AU2023248067B2 - Cereblon ligands and bifunctional compounds comprising the same - Google Patents

Abstract

#$%^&*AU2023248067B220250904.pdf##### ABSTRACT The description relates to cereblon E3 ligase binding compounds, including bifunctional compounds comprising the same, which find utility as modulators of targeted ubiquitination, especially inhibitors of a variety of polypeptides and other proteins which are degraded and/or otherwise inhibited by bifunctional compounds according to the present disclosure. In particular, the description provides compounds, which contain on one end a ligand which binds to the cereblon E3 ubiquitin ligase and on the other end a moiety which binds a target protein such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of that protein. Compounds can be synthesized that exhibit a broad range of pharmacological activities consistent with the degradation/inhibition of targeted polypeptides of nearly any type. ABSTRACT 10 Oct 2023 The description relates to cereblon E3 ligase binding compounds, including bifunctional compounds comprising the same, which find utility as modulators of targeted ubiquitination, especially inhibitors of a variety of polypeptides and other proteins which are degraded and/or otherwise inhibited by bifunctional compounds according to the present disclosure. In particular, the description provides compounds, which contain on one end a ligand which binds to the 2023248067 cereblon E3 ubiquitin ligase and on the other end a moiety which binds a target protein such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of that protein. Compounds can be synthesized that exhibit a broad range of pharmacological activities consistent with the degradation/inhibition of targeted polypeptides of nearly any type.

Description

CEREBLON LIGANDS AND ANDBIFUNCTIONAL BIFUNCTIONALCOMPOUNDS COMPOUNDS 2023248067 10 Oct 2023

CEREBLON LIGANDS COMPRISING THE COMPRISING THE SAME SAME CROSS-REFERENCETO CROSS-REFERENCE TORELATED RELATEDAPPLICATIONS APPLICATIONS

[0001] This This

[0001] application application is a divisional is a divisional application application of Australian of Australian Patent Application Patent Application No. No. 2019251223filed 2019251223 filed on on 9 April 9 April 2019,2019, of which of which is the Australian is the Australian national national phase phase entry entry of an of an International Application International claiming priority Application claiming priority to to U.S. U.S. Non-Provisional ApplicationNo. Non-Provisional Application No. 15/953108, 15/953108,

filed filed 13 13 April 2018, 2018, which whichisisa aContinuation-in-Part Continuation-in-Partof of U.S.Non-Provisional U.S. Non-Provisional Application Application No. No.

14/686,640, filed 14/686,640, filed 1414April April2015, 2015, thatthat claims claims priority priority to U.S. to U.S. Provisional Provisional Application Application No. No. 61/979,351, filed 14 61/979,351, filed 14 April April 2014, 2014, and and aa Continuation-in-Part Continuation-in-Part of ofU.S. U.S. Non-Provisional Non-ProvisionalApplication Application No. 14/792,414, No. 14/792,414,filed filed6 6July July 2015, 2015, thatthat claims claims priority priority to U.S. to U.S. Provisional Provisional Application Application No. No. 61/979,351, filed 61/979,351, filed 14 14 April April 2014, 2014, and andU.S. U.S.Provisional ProvisionalApplication Application No.No. 62/171,090, 62/171,090, filed filed 4 June 4 June

2015,all 2015, all ofofwhich whichareare incorporated incorporated herein herein by reference by reference in theirinentirety. their entirety.

BYREFERENCE INCORPORATIONBY INCORPORATION REFERENCE

[0002] U.S. U.S.

[0002] Patent Patent Application Application Serial Serial No. 15/230,354, filed filed No. 15/230,354, on August on August 5, 2016, 5, 2016, published published as as U.S. Patent U.S. Patent Application Application Publication PublicationNo. No.2017/0065719; 2017/0065719; and and U.S. U.S. Patent Patent Application Application SerialSerial No. No. 15/801,243, filed 15/801,243, filed on on 11 November November 2017; 2017; and U.S. and U.S. Patent Patent Application Application 15/206,497 15/206,497 filed 11filed July11 July 2016; and 2016; andU.S. U.S.Patent PatentApplication Application 15/209,648 15/209,648 filed filed 13 13 JulyJuly 2016; 2016; and and U.S. U.S. Patent Patent Application Application

Serial No. Serial No. 15/730,728, filed filed on on October 11, 2017; October 11, 2017; U.S. U.S. Patent Patent Application Application Serial Serial No. No. 15/829,541, 15/829,541, filed filed on on December December 1,1,2017; 2017;U.S. U.S.Patent PatentApplication Application SerialNo.No. Serial 15/881,318, 15/881,318, filed filed on on January January 26, 26,

2018; and 2018; andU.S. U.S. Patent PatentApplication ApplicationSerial SerialNo. No.14/686,640, 14/686,640, filed filed on on April April 14,14, 2015, 2015, published published as as U.S. Patent U.S. Patent Application Application Publication PublicationNo. No.2015/0291562; 2015/0291562; and and U.S. U.S. Patent Patent Application Application SerialSerial No. No. 14/792,414, filed 14/792,414, filed ononJuly July6, 6, 2015, 2015, published published as Patent as U.S. U.S. Patent Application Application Publication Publication No. No. 2016/0058872;andand 2016/0058872; U.S.U.S. Patent Patent Application Application SerialSerial No. 14/371,956, No. 14/371,956, filed onfiled July on 11, July 2014,11, 2014, published as published as U.S. U.S. Patent Patent Application Application Publication Publication No. No. 2014/0356322; 2014/0356322;andand U.S. U.S. Patent Patent Application Application

Serial No. Serial 15/074,820, filed No. 15/074,820, filed on March18, on March 18,2016, 2016,published publishedas as U.S. U.S. Patent Patent Application Application

Publication No. Publication No. 2016/0272639; 2016/0272639;andand U.S. U.S. Patent Patent Application Application Serial Serial No. No. 15/885,671, 15/885,671, filed filed on 31on 31 January 2018, January 2018, are are incorporated incorporatedherein hereinbybyreference referenceinintheir their entirety. entirety. Furthermore, Furthermore, all all references references cited hereinare cited herein areincorporated incorporated by reference by reference hereinherein in entirety. in their their entirety.

FIELD OF THE THE INVENTION INVENTION 2023248067 10 Oct 2023

FIELD OF The description

[0003] The description

[0003] provides provides imide-based imide-based compounds, compounds, including including bifunctional bifunctional compounds compounds

comprising thesame, comprising the same,and andassociated associatedmethods methodsof of use. use. TheThe bifunctional bifunctional compounds compounds are useful are useful as as modulators ofoftargeted modulators targetedubiquitination, ubiquitination, especially especially with withrespect respecttotoa avariety variety ofofpolypeptides polypeptidesandand other proteins, proteins, which whicharearedegraded degraded and/or and/or otherwise otherwise inhibited inhibited by bifunctional by bifunctional compounds compounds

according according toto thepresent the present disclosure. disclosure.

BACKGROUND BACKGROUND

[0004] Most

[0004] Most small small molecule molecule drugs bindbind drugs enzymes enzymes or receptors or receptors in tight andand in tight well-defined well-defined

pockets.On On pockets. the the other other hand,hand, protein-protein protein-protein interactions interactions are notoriously are notoriously difficult todifficult to target target using using small molecules moleculesdue duetototheir theirlarge largecontact contactsurfaces surfacesandand thethe shallow shallow grooves grooves or flat or flat interfaces interfaces

involved. E3E3 ubiquitin ubiquitin ligases ligases (of(of which which hundreds hundreds are known are known in confer in humans) humans) confer substrate substrate specificity forubiquitination, specificity for ubiquitination,andand therefore, therefore, are attractive are more more attractive therapeutic therapeutic targets targets than than general general

proteasomeinhibitors proteasome inhibitors due duetoto their their specificity specificity for for certain certainprotein proteinsubstrates. substrates.The Thedevelopment of development of

ligands of of E3 E3ligases ligases has hasproven proven challenging, challenging, in part in part due due to fact to the the fact that that they they must must disrupt disrupt

protein-protein interactions. protein-protein interactions. However, However, recent recent developments developments have provided have provided specific specific ligands ligands whichbind which bind to to these these ligases. ligases. For For example, example, since since the the discovery discovery ofthe of nutlins, nutlins, the first first small small molecule molecule E3 ligaseinhibitors, E3 ligase inhibitors,additional additional compounds compounds have have been been that reported reported targetthat target E3 E3 ligases ligases but the fieldbut the field

remains underdeveloped. remains underdeveloped.

[0005] OneOne

[0005] E3 ligase E3 ligase withwith therapeutic therapeutic potential the von potential isis the von Hippel-Lindau Hippel-Lindau (VHL) tumor (VHL)tumor suppressor. VHL suppressor. VHL comprises comprises the substrate the substrate recognition recognition subunit/E3 subunit/E3 ligaseligase complex complex VCB, which VCB, which

includes elongins BB and and C, C, and andaa complex complexincluding includingCullin-2 Cullin-2andand Rbxl. Rbx1. TheThe primary primary substrate substrate of of

VHLisisHypoxia VHL Hypoxia Inducible Inducible Factor Factor la (HIF-la), 1 (HIF-1), a transcription a transcription factor factor thatthat upregulates upregulates genes genes suchsuch

as the pro-angiogenic as the pro-angiogenic growth growth factor factor VEGF VEGFand and the blood the red red blood cell inducing cell inducing cytokine cytokine

erythropoietin in erythropoietin in response response to to low low oxygen levels. We oxygen levels. generatedthe We generated thefirst first small small molecule ligands of molecule ligands of Von Hippel Von Hippel Lindau Lindau(VHL) (VHL) to to thethe substraterecognition substrate recognition subunit subunit of of the the E3 ligase, VCB, E3 ligase, an VCB, an

important target inincancer, important target cancer,chronic chronic anemia anemia and ischemia, and ischemia, and obtained and obtained crystal structures crystal structures

confirming that the confirming that the compound compound mimics mimics the the binding binding mode mode of theof the transcription transcription factor factor HIF-la, HIF-1, the the major substrate major substrate of of VHL. VHL.

[0006] Cereblon

[0006] Cereblon is a protein is a protein thatthat in humans in humans is encoded is encoded byCRBN by the the CRBN gene. gene. CRBN CRBN orthologs orthologs are are highly conserved conservedfrom from plantsto to plants humans, humans, which which underscores underscores its physiological its physiological importance. importance.

2

Cereblon forms formsananE3E3 ubiquitinligase ligasecomplex complex withwith damaged DNA binding protein protein 1 (DDB1), 2023248067 10 Oct 2023

Cereblon ubiquitin damaged DNA binding 1 (DDB1),

Cullin-4A (CUL4A), Cullin-4A (CUL4A), andand regulator regulator of of cullins1 (ROC1). cullins 1 (ROC1). ThisThis complex complex ubiquitinates ubiquitinates a number a number of of other other proteins. proteins.Through Through aa mechanism whichhas mechanism which hasnot notbeen beencompletely completelyelucidated, elucidated, cereblon cereblon ubquitination of ubquitination of target target proteins proteins results resultsininincreased increased levels levelsoffibroblast of fibroblastgrowth growth factor factor8 8(FGF8) (FGF8)

and fibroblast growth and fibroblast growthfactor factor 1010(FGF10). (FGF10).FGF8FGF8 in regulates in turn turn regulates a number a number of developmental of developmental

processes,such processes, such as as limb limb and and auditory auditory vesicle vesicle formation. formation. The net The net result is result is that that this this ubiquitin ubiquitin ligase ligase complex complex isis important importantfor forlimb limboutgrowth outgrowthin in embryos. embryos. In In thethe absence absence of of cereblon, cereblon, DDB1 DDB1 formsforms a a complex withDDB2 complex with DDB2thatthat functions functions as as a DNA a DNA damage-binding damage-binding protein. protein.

Thalidomide, which

[0007] Thalidomide,

[0007] beenapproved hasbeen whichhas forthe approvedfor thetreatment numberof of of a anumber treatment of immunological indications,hashas immunological indications, also also beenbeen approved approved for thefor the treatment treatment of neoplastic of certain certain neoplastic diseases, including diseases, including multiple multiple myeloma. myeloma. InInaddition additiontotomultiple multiplemyeloma, myeloma, thalidomide thalidomide and and several several

of its analogs of its arealso analogs are alsocurrently currently under under investigation investigation for usefor in use in treating treating a variety a variety of other of other types of types of

cancer. cancer. While the precise While the precise mechanism mechanism of of thalidomide's thalidomide's anti-tumor anti-tumor activity activity is is still emerging, still emerging,ititis is knowntotoinhibit known inhibit angiogenesis. angiogenesis. Recent Recentliterature literature discussing the biology discussing the of the biology of the imides imides includes includes Lu Lu

et et al alScience Science 343, 343, 305 305 (2014) (2014) and Kr6nkeetetal and Krönke al Science Science 343, 343, 301 301 (2014). (2014).

[0008] Significantly,

[0008] Significantly, thalidomide thalidomide and analogs and its its analogs e.g. e.g. pomolinamiode pomolinamiode and lenalinomide, and lenalinomide, are are knowntotobind known bindcereblon. cereblon.These These agents agents bind bind to to cereblon,altering cereblon, alteringthe the specificity specificity of of the the complex to complex to

induce the ubiquitination induce the ubiquitination and and degradation degradationofofIkaros Ikaros(IKZF1) (IKZF1) andand Aiolos Aiolos (IKZF3), (IKZF3), transcription transcription

factors factors essential essentialfor formultiple multiplemyeloma growth.Indeed, myeloma growth. Indeed, higher higher expression expression of cereblon of cereblon has has beenbeen

linked to ananincrease linked to increasein in efficacy efficacy of imide of imide drugsdrugs in theintreatment the treatment of multiple of multiple myeloma. myeloma.

[0009] An ongoing

[0009] An ongoing need in need exists exists in the the art for art for effective effective treatments treatments for disease, for disease, especially especially

hyperplasias and hyperplasias andcancers, cancers, such suchasasmultiple multiple myeloma. myeloma. However, However, non-specific non-specific effects,effects, and theand the inability inability to to target target and modulate modulatecertain certainclasses classesof of proteins proteins altogether,such altogether, such as transcription as transcription

factors, factors, remain as obstacles remain as obstacles to to the the development development ofof effectiveanti-cancer effective anti-canceragents. agents.AsAssuch, such,small small moleculetherapeutic molecule therapeutic agents agents that that leverage leverage or potentiate or potentiate cereblon's cereblon's substratesubstrate specificity specificity and, at theand, at the same time,areare"tunable" same time, "tunable" suchsuch that that a wide a wide range range of of protein protein classes classes can be targetted can be targetted and and modulated modulated with with specificity specificity wouldwould beuseful be very very as useful as a therapeutic. a therapeutic.

BRIEF SUMMARY BRIEF OFTHE SUMMARY OF THEINVENTION INVENTION The present

[0010] The present

[0010] disclosure disclosure describes describes bifunctional bifunctional compounds compounds which function recruitto function which to recruit endogenousproteins endogenous proteinstotoananE3E3Ubiquitin Ubiquitin Ligase Ligase forfor degradation, degradation, andand methods methods of using of using the the same. same.

3

In particular, the present particular, the present disclosure disclosure provides providesbifunctional or or bifunctional proteolysis targeting chimeric 2023248067 10 Oct 2023

In proteolysis targeting chimeric

(PROTAC) which which compounds, (PROTAC) compounds, find utility find utility as modulators as modulators of targeted of targeted ubiquitination ubiquitination of a variety of a variety

of polypeptides of andother polypeptides and otherproteins, proteins, which whicharearethen then degraded degraded and/or and/or otherwise otherwise inhibited inhibited by by the the bifunctional compounds bifunctional compounds asas described described herein.AnAn herein. advantage advantage of the of the compounds compounds provided provided hereinherein is is that aa broad that broadrange range of pharmacological of pharmacological activities activities is possible, is possible, consistent consistent with with the the degradation/inhibition of degradation/inhibition of targeted targeted polypeptides polypeptides from fromvirtually virtuallyany anyprotein protein classor or class family.In family. In addition, the addition, the description description provides methodsofof provides methods using using an an effectiveamount effective amount of the of the compounds compounds as as described herein described herein for for the the treatment treatment or or amelioration ameliorationofofa adisease diseasecondition, condition,such suchas as cancer,e.g., cancer, e.g., multiple myeloma. multiple myeloma.

[0011] As As

[0011] such, such, in one in one aspect thethe aspect disclosure providesnovel disclosureprovides compounds imide-basedcompounds novelimide-based as as described herein. described herein.

[0012] In anInadditional

[0012] an additional aspect, aspect, the the disclosure disclosure provides provides bifunctional bifunctional or or PROTAC PROTAC compounds, compounds,

whichcomprise which compriseananE3E3 Ubiquitin Ubiquitin Ligase Ligase binding binding moiety moiety (i.e.,a aligand (i.e., ligandfor forananE3E3Ubiquitin UbiquitinLigase Ligase or "ULM" or group), "ULM" group), andand a moiety a moiety thatthat binds binds a target a target protein protein (i.e.,a aprotein/polypeptide (i.e., protein/polypeptidetargeting targeting ligand or or "PTM" "PTM" group) group) suchsuch thatthat the the target target protein/polypeptide protein/polypeptide is placed is placed in proximity in proximity to the to the

ubiquitinligase ubiquitin ligasetotoeffect effectdegradation degradation (and (and inhibition) inhibition) ofprotein. of that that protein. In a preferred In a preferred embodiment, embodiment, the ULM the ULM is is a a cereblon cereblon E3 E3 Ubiquitin Ubiquitin Ligase Ligase binding binding moiety moiety (i.e., (i.e., a "CLM"). a "CLM"). For example, For example, the the structure structure of of the thebifunctional bifunctionalcompound can be compound can bedepicted depicted as: as:

PTM CLM

[0013] TheThe

[0013] respectivepositions respective the PTM of the positions of PTM and CLM andCLM moieties as as moieties well their number wellasastheir as number as illustrated illustratedherein hereinisis provided providedbybyway way of of example only and example only andis is not not intended to to limit limit the thecompounds compounds

in any any way. way.As As would would be understood be understood by the by the skilled skilled artisan, artisan, the bifunctional the bifunctional compounds compounds as as described herein described hereincan canbe be synthesized synthesized such such thatnumber that the the number and position and position of the respective of the respective

functionalmoieties functional moietiescancan be varied be varied as desired. as desired.

[0014] In In

[0014] certainembodiments, certain thethe embodiments, bifunctional compound bifunctional compound further comprisesa achemical furthercomprises chemical linker ("L"). linker ("L"). InInthis thisexample, example, the the structure structure ofbifunctional of the the bifunctional compound compound can be can be depicted as:depicted as:

PTM L CLM where PTM where PTMis is a protein/polypeptide a protein/polypeptide targeting targeting moiety, moiety, L ais linker, L is a linker,andand CLMCLM is a is a cereblon cereblon E3 E3 ubiquitinligase ubiquitin ligasebinding binding moiety. moiety.

4

[0015] In certain

[0015] In certain preferred preferred embodiments, embodiments, the E3the E3 Ubiquitin Ubiquitin Ligase Ligase is cereblon. is cereblon. As such,As in such, in certain certain additional additional embodiments, theCLM embodiments, the CLM of the of the bifunctional bifunctional compound compound comprises comprises chemistries chemistries

such as imide, such as imide, amide, amide,thioamide, thioamide,thioimide thioimide derived derived moieties. moieties. In additional In additional embodiments, embodiments, the the CLM comprises CLM comprises a phthalimido a phthalimido group group or an or an analog analog or derivative or derivative thereof.thereof. In still additional In still additional

embodiments, theCLMCLM embodiments, the comprises comprises a phthalimido-glutarimide a phthalimido-glutarimide group group or or an oranalog an analog or derivative derivative

thereof. In thereof. In still still other other embodiments, theCLM embodiments, the CLM comprises comprises a member a member of the of the consisting group group consisting of of thalidomide, lenalidomide, thalidomide, lenalidomide, pomalidomide, pomalidomide,andand analogs analogs or or derivativesthereof. derivatives thereof.

[0016] In certain

[0016] In certain embodiments, embodiments, the compounds the compounds as described as described herein herein comprise comprise multiplemultiple CLMs, CLMs, multiple PTMs, multiple multiplechemical PTMs, multiple chemicallinkers linkersororaacombination combinationthereof. thereof.

[0017] In In

[0017] anyany aspect aspect or embodiment or embodiment described described herein, herein, the the ULM ULM (ubiquitination (ubiquitination ligase ligase

modulator) can modulator) canbebeVon Von Hippel-Lindau Hippel-Lindau E3 ubiquitin E3 ubiquitin ligase ligase (VHL)(VHL) bindingbinding moiety moiety (VLM), (VLM), or a or a cereblon cereblon E3 ubiquitin ligase E3 ubiquitin ligasebinding bindingmoiety moiety(CLM), (CLM), or or aa mouse double minute mouse double minute 22 homolog homolog (MDM2) (MDM2) E3E3 ubiquitinligase ubiquitin ligase binding binding moiety moiety (MLM), (MLM),ororananIAP IAP E3 E3 ubiquitinligase ubiquitin ligase binding binding moiety (i.e., moiety (i.e., aa "ILM"). "ILM").In any In aspect any aspect or embodiments or embodiments describeddescribed herein, herein, the the bifunctional bifunctional

compound includes compound includes at least at least oneone additional additional E3 ligase E3 ligase binding binding moiety moiety selected selected from from the the group group

consisting consistingofofVLM, VLM, VLM', CLM,CLM', VLM', CLM, CLM', MLM, MLM, MLM', MLM', ILM, ILM', ILM, ILM', or a combination or a combination thereof. thereof.

For example, For example, there there can can beleast be at at least 1, 2,1, 3, 2, 4,3, or 4, 5oradditional 5 additional E3 ligase E3 ligase binding binding moieties. moieties.

[0018] In anInadditional

[0018] an additional aspect, aspect, the the description description provides provides therapeutic therapeutic compositions compositions comprising comprising

an effective amount an effective amount ofofa acompound compound as described as described herein herein or form or salt salt form thereof, thereof, and aand a

pharmaceutically acceptable pharmaceutically acceptablecarrier. carrier. The Thetherapeutic therapeutic compositions compositionsmodulate modulate protein protein degradation degradation

in aa patient patient or orsubject, subject,for forexample, example, an an animal animal such as aa human, such as andcan human, and canbebeused used forfor treatingoror treating

ameliorating disease ameliorating disease states states or or conditions conditions which are modulated which are modulatedthrough through thethe degraded degraded protein. protein. In In certain embodiments, certain thetherapeutic embodiments, the therapeutic compositions compositionsasasdescribed describedherein hereinmay may be be used used to effectuate to effectuate

the degradation the degradation of of proteins proteins of interest of interest for treatment for the the treatment or amelioration or amelioration of a disease, of a disease, e.g., e.g., cancer. cancer. In yet In yet another another aspect, aspect, the the present present disclosure disclosure provides provides a amethod methodof of ubiquitinating/ ubiquitinating/ degrading degrading a a target protein target protein in in aa cell. cell.InIn certain certainembodiments, embodiments, the the method comprises administering method comprises administering aa bifunctional compound bifunctional compound as as described described herein herein comprising comprising an and an CLM CLM and preferably a PTM, a PTM, preferably linked linked through aa linker through linker moiety, as as otherwise described herein, otherwise described herein, wherein whereinthe the CLM CLM is coupled is coupled to the to the PTMPTM

and wherein and wherein the the CLM CLM recognizes recognizes a ubiquitin a ubiquitin pathway pathway protein protein (e.g.,an an (e.g., ubiquitinligase, ubiquitin ligase, preferably an preferably an E3 E3ubiquitin ubiquitinligase ligasesuch suchas,as,e.g., e.g.,cereblon) cereblon)andand thethe PTMPTM recognizes recognizes the target the target

proteinsuch protein suchthat thatdegradation degradation oftarget of the the target protein protein will when will occur occur thewhen targetthe targetis protein protein placed inis placed in proximity to the ubiquitin ligase, thus resulting in degradation/inhibition of the effects of the 06 Aug 2025 target protein and the control of protein levels. The control of protein levels afforded by the present disclosure provides treatment of a disease state or condition, which is modulated through the target protein by lowering the level of that protein in the cells of a patient.

[0019] In an additional aspect, the description provides a method for assessing (i.e., determining and/or measuring) a CLM’s binding affinity. In certain embodiments, the method comprises providing a test agent or compound of interest, for example, an agent or compound 2023248067

having an imide moiety, e.g., a phthalimido group, phthalimido-glutarimide group, derivatized thalidomide, derivatized lenalidomide or derivatized pomalidomide, and comparing the cereblon binding affinity and/or inhibitory activity of the test agent or compound as compared to an agent or compound known to bind and/or inhibit the activity of cereblon.

[0020] In still another aspect, the description provides methods for treating or emeliorating a disease, disorder or symptom thereof in a subject or a patient, e.g., an animal such as a human, comprising administering to a subject in need thereof a composition comprising an effective amount, e.g., a therapeutically effective amount, of a compound as described herein or salt form thereof, and a pharmaceutically acceptable carrier, wherein the composition is effective for treating or ameliorating the disease or disorder or symptom thereof in the subject.

[0021] In another aspect, the description provides methods for identifying the effects of the degradation of proteins of interest in a biological system using compounds according to the present disclosure.

[0021a] In still another aspect, the description provides a compound selected from the group consisting of

6a

06 Aug 2025

Ex. # Chemical Structure

1 2023248067

2

3

4

5

6a

6b

2023248067 06 Aug 2025

9 8 7 6

6b

6c

2023248067 06 Aug 2025

14 13 12 11 10

6c p9

2023248067 06 Aug 2025

18 17 16 15

6d

6e

2023248067 06 Aug 2025

23 22 21 20 19

6e

6f

2023248067 06 Aug 2025

28 27 26 25 24

6f

6g

29 06 Aug 2025

30 2023248067

31

or a pharmaceutically acceptable salt thereof. The description further provides a composition comprising an effective amount of the above-mentioned compound and a pharmaceutically acceptable carrier.

[0021b] In still another aspect, the description provides use of an effective amount of the above-mentioned compound of in the manufacture of a medicament for treating a disease or disorder in a subject, wherein the disease or disorder is associated with the accumulation and/or aggregation of the target protein. The description further provides use of an effective amount of the above-mentioned compound in the manufacture of a medicament for inducing degradation of a target protein or for treating cancer, wherein the compound effectuates degradation of the target protein, or wherein the compound effectuates for the treatment or alleviation of at least one symptom of cancer in the patient.

[0021c] In still another aspect, the description provides a method for treating a disease or disorder in a subject, wherein the disease or disorder is associated with the accumulation and/or

6g

6h

aggregation of the target protein, said method comprising administering to the subject an 06 Aug 2025

effective amount of the above-mentioned compound, or a pharmaceutically acceptable thereof. The description further provides a method for inducing degradation of a target protein in a cell, the method comprising administering an effective amount of the above-mentioned compound to the cell, wherein the compound effectuates degradation of the target protein. Furthermore, the description further provides a method for treating cancer, said method comprising administering the above-mentioned compound to a patient in need thereof, wherein the compound effectuates 2023248067

for the treatment or alleviation of at least one symptom of cancer in the patient.

[0022] The preceding general areas of utility are given by way of example only and are not intended to be limiting on the scope of the present disclosure and appended claims. Additional advantages associated with the compositions, methods, and processes of the present disclosure will be appreciated by one of ordinary skill in the art in light of the instant claims, description, and examples. For example, the various aspects and embodiments of the invention may be utilized in numerous combinations, all of which are expressly contemplated by the present description. These additional advantages and embodiments are expressly included within the scope of the present disclosure. The publications and other materials used herein to illuminate the background of the invention, and in particular cases, to provide additional details respecting the practice, are incorporated by reference.

[0022a] A reference herein to a patent document or any other matter identified as prior art, is not to be taken as an admission that the document or other matter was known or that the information it contains was part of the common general knowledge as at the priority date of any of the claims.

6h

BRIEF DESCRIPTION DESCRIPTION OF OF THE THE DRAWINGS 2023248067 10 Oct 2023

BRIEF DRAWINGS

[0023] TheThe

[0023] accompanying accompanying drawings, drawings, which which are incorporated and and intointo are incorporated form form a part of of a part thethe

specification, specification, illustrate illustrateseveral severalembodiments embodiments ofofthe thepresent presentdisclosure disclosureand, and,together together with with the the

description, description, serve to to explain explain the the principles principles of ofthe the invention. invention.TheThe drawings drawings are only are only for the for the

purposeofof purpose illustratingan an illustrating embodiment embodiment of the of the invention invention andtoare and are not be not to be as construed construed as limiting limiting the the invention. Further objects, invention. Further objects, features features and advantages ofofthe and advantages the invention inventionwill will become becomeapparent apparent from from

the following the detailed description following detailed description taken taken in in conjunction conjunction with withthe theaccompanying accompanying figures figures showing showing

illustrative embodiments illustrative embodiments of invention, of the the invention, in which: in which:

[0024] Figure

[0024] Figure 1A 1B. 1A and and Illustration 1B. Illustration of general of general principlefor principle forPROTAC PROTAC function. function. (A) (A) ExemplaryPROTACs Exemplary PROTACs comprise comprise a protein a protein targeting targeting moiety moiety (PTM;shaded (PTM; darkly darkly shaded rectangle), rectangle), a a ubiquitin ligase ubiquitin ligase binding binding moiety (ULM;lightly moiety (ULM; lightlyshaded shaded triangle),and triangle), and optionally optionally aa linker linker moiety (L; moiety (L;

black line) black line) coupling or tethering coupling or tethering the the PTM to the PTM to the ULM. ULM.(B) (B) Illustrates Illustrates thethe functional functional useuse of of thethe

PROTACs PROTACs as described as described herein. herein. Briefly, Briefly, thethe ULM ULM recognizes recognizes and binds and binds to a to a specific specific E3 Ubiquitin E3 Ubiquitin

Ligase,and Ligase, andthethe PTMPTM bindsbinds and recruits and recruits a targeta protein target protein bringing bringing it into it into close close proximity proximity to the E3 to the E3 Ubiquitin Ligase. Typically, Ubiquitin Ligase. Typically, the the E3 Ubiquitin E3 Ubiquitin LigaseLigase is complexed is complexed with with an E2 an E2 ubiquitin ubiquitin-

conjugating protein, and conjugating protein, andeither either alone aloneororvia viathetheE2E2 protein protein catalyzes catalyzes attachment attachment of ubiquitin of ubiquitin

(dark circles) to (dark circles) to aa lysine lysine on on the the target target protein protein via via an an isopeptide isopeptide bond. Thepoly-ubiquitinated bond. The poly-ubiquitinated protein(far protein (farright) right)isisthen thentargeted targetedforfor degration degration by proteosomal by the the proteosomal machinery machinery of of the cell. the cell.

DETAILED DESCRIPTION DETAILED DESCRIPTION The following

[0025] The following

[0025] is a detailed is a detailed description description provided provided to aid those those skilled to aidskilled in the in the art in art in practicing the practicing the present present disclosure. disclosure. Those of ordinary skill Those of skill in inthe theart artmay may make modifications and make modifications and variations variations in in the the embodiments describedherein embodiments described hereinwithout withoutdeparting departingfrom from thethe spiritororscope spirit scopeofofthe the present disclosure. present disclosure. All All publications, publications, patent patent applications, applications, patents, patents, figures figures and and other otherreferences references mentioned mentioned herein herein are are expressly expressly incorporated incorporated by reference by reference in their entirety. in their entirety.

[0026] Presently

[0026] Presently described described are compositions are compositions and methods and methods thattorelate that relate to the surprising the surprising and and unexpecteddiscovery unexpected discoverythat thatananE3E3Ubiquitin Ubiquitin Ligase Ligase protein, protein, e.g.,cereblon, e.g., cereblon,ubiquitinates ubiquitinatesa target a target protein once protein once itit and andthe thetarget targetprotein proteinare areplaced placed in in proximity proximity by aby a bifunctional bifunctional or chimeric or chimeric

construct that that binds the E3 binds the E3Ubiquitin UbiquitinLigase Ligase protein protein and and the target the target protein. protein. Accordingly Accordingly the the

present disclosure present disclosure provides providessuch suchcompounds compounds and compositions and compositions comprising comprising an E3 Ubiquintin an E3 Ubiquintin

Ligase binding Ligase bindingmoiety moiety ("ULM") ("ULM") coupled coupled to a protein to a protein target target binding binding moiety moiety ("PTM"),("PTM"), which which result in in the the ubiquitination ubiquitination of of aa chosen target protein, protein, which leads to to degradation degradation of ofthe the target target 2023248067 10 Oct 2023 result chosen target which leads protein by protein by the the proteasome (see proteasome(see Figures Figures 1A 1B). 1A and and The 1B).present The present disclosure disclosure also provides also provides a a library of compositions library of compositions and and the thereof. the use use thereof.

[0027] In In

[0027] the present aspects, the certainaspects, certain present disclosure provides compounds disclosure provides compounds which comprisea whichcomprise a ligand, e.g., aa small ligand, e.g., smallmolecule molecule ligand ligand (i.e., (i.e., having having a molecular a molecular weight weight of below of below 2,000, 2,000, 1,000, 500, 1,000, 500,

or 200 Daltons), which 200 Daltons), whichisis capable capable of of binding binding to to aa ubiquitin ligase, ligase,such such as asIAP, IAP, VHL, MDM2, VHL, MDM2, or or

cereblon. Thecompounds cereblon. The compoundsalsoalso comprise comprise a moiety a moiety that that is capable is capable of binding of binding to target to target protein, protein, in in

such such aaway way that that thethe target target protein protein is placed is placed in proximity in proximity to thetoubiquitin the ubiquitin ligase ligase to effect to effect

degradation (and/or degradation (and/or inhibition) inhibition) of of that that protein. protein. Small Small molecule molecule can can mean, mean, in addition in addition to theto the above, thatthethe above, that molecule molecule is non-peptidyl, is non-peptidyl, that is,that is,not it is it generally is not generally consideredconsidered a peptide, e.g., a peptide, e.g., comprises fewerthan comprises fewer than4, 4,3,3,oror2 2amino amino acids. acids. In accordance In accordance withpresent with the the present description, description, the the

PTM, ULM PTM, ULMor or PROTAC PROTAC molecule molecule can can be abesmall a small molecule. molecule.

[0028] Unless

[0028] Unless otherwise otherwise defined, defined, all technical and and all technical scientific scientific terms terms used used herein herein have the the have samesame

meaningasascommonly meaning commonly understood understood by oneby of one of ordinary ordinary skill inskill the in artthe to art to this which which this invention invention

belongs. The belongs. Theterminology terminologyused used in in thethe descriptionis isfor description fordescribing describingparticular particular embodiments embodiments only only

andisis not and not intended intendedto to be be limiting limiting of the of the invention. invention.

[0029] WhereWhere

[0029] a range a range of values of values is provided, is provided, it is it is understood understood that intervening that each each intervening value,value, to to the tenth the tenth ofofthe theunit unitofofthe thelower lower limit limit unless unless the the context context clearly clearly dictates dictates otherwise otherwise (such as(such in theas in the case of of aa group containinga anumber group containing numberof of carbon carbon atoms atoms in which in which case case each carbon each carbon atom number atom number

falling falling within within the the range is provided), range is provided), between theupper between the upperand and lower lower limit limit of of thatrange that range andand anyany

other stated other stated or or intervening intervening value in in that that stated stated range range is is encompassed withinthe encompassed within theinvention. invention.The The upper and upper andlower lowerlimits limitsofofthese thesesmaller smallerranges ranges maymay independently independently be included be included in the in the smaller smaller

rangesisisalso ranges alsoencompassed encompassedwithinwithin the invention, the invention, subject subject to any specifically to any specifically excluded excluded limit in the limit in the stated stated range. range. Where thestated Where the stated range range includes includesone oneororboth bothofofthe thelimits, limits, ranges rangesexcluding excludingeither either bothofofthose both thoseincluded included limits limits are are alsoalso included included in theininvention. the invention.

[0030] terms terms The following

[0030] The following are used used are to to describe describe the present the present invention. invention. In instances where where In instances a a term is term is not not specifically specificallydefined defined herein, herein,that thatterm termisisgiven givenananart-recognized art-recognizedmeaning by those meaning by those of of ordinaryskill ordinary skillapplying applying that that term term in context in context to use to its its use in describing in describing the present the present invention. invention.

[0031] "a" "a" The articles

[0031] The articles and and "an""an" as used as used herein and and herein in the in the appended appended claims claims are used are used herein herein to to refer to refer to one or to one or to more more than thanone one(i.e., (i.e., to to at at least least one) one) of the the grammatical objectofofthe grammatical object thearticle article

8 unless the the context context clearly clearly indicates indicates otherwise. otherwise. ByBywayway of example, "an element" means one 2023248067 10 Oct 2023 unless of example, "an element" means one element or more element or morethan thanone oneelement. element.

The phrase

[0032] The phrase

[0032] "and/or," "and/or," as used hereinherein as used the specification in theinspecification and in the claims, andthein claims, should should be be understood understood totomean mean "either "either or both" or both" ofelements of the the elements so conjoined, so conjoined, i.e., elements i.e., elements that are that are

conjunctively present conjunctively present in in some somecases casesand anddisjunctively disjunctivelypresent presentininother othercases. cases.Multiple Multipleelements elements listed listed with with "and/or" "and/or" should should be construed construed in in the the same fashion, i.e., same fashion, i.e., "one "one or ormore" of the more" of the elements elements

so conjoined. conjoined. Other Otherelements elements maymay optionally optionally be present be present other other thanelements than the the elements specifically specifically

identified identified by the "and/or" by the "and/or" clause, clause, whether whetherrelated relatedor orunrelated unrelated to to those those elements elements specifically specifically

identified. identified.Thus, Thus,asasa anon-limiting non-limitingexample, example,a areference referencetoto"A "A and/or and/or B", B", when used in when used in conjunction with open-ended conjunction with open-endedlanguage language such such as "comprising" as "comprising" can refer, can refer, in one in one embodiment, embodiment, to A to A

only (optionally (optionally including elements elements other other than than B); B); in in another another embodiment, embodiment, to to B B only only (optionally (optionally

including elements elementsother otherthan than A);A); in yet in yet another another embodiment, embodiment, to bothtoA both and BA(optionally and B (optionally including otherelements); including other elements); etc.etc.

[0033] As used

[0033] hereinherein As used the specification in theinspecification and in theinclaims, and the claims, "or" should "or" should be understood be understood to to have the have the same samemeaning meaning as "and/or" as "and/or" as defined as defined above. above. For For example, example, when separating when separating items items in a in a list, list, "or" "or" or or "and/or" shallbebeinterpreted "and/or" shall interpreted as being as being inclusive, inclusive, i.e., i.e., the inclusion the inclusion of at of at one, least leastbut one, but also includingmore also including more thanthan one, one, of a of a number number or list or of list of elements, elements, and, optionally, and, optionally, additional additional unlisted unlisted items. Onlyterms items. Only terms clearly clearly indicated indicated to thetocontrary, the contrary, such assuch "onlyas "only one of or one of orone"exactly "exactly of," or, one of," or,

whenused when usedininthe theclaims, claims,"consisting "consistingof," of," will will refer refer to to the the inclusion inclusion of of exactly exactly one one element of aa element of

numberororlist number list of of elements. elements. In In general, general, the the term term "or" "or" as as used used herein herein shall shall only only be be interpreted interpreted as as indicating exclusive indicating exclusive alternatives alternatives (i.e.,"one (i.e., "one or the or the other other butboth") but not not both") when preceded when preceded by terms ofby terms of

exclusivity, suchasas"either," exclusivity, such "either,""one "one of,"of," "only "only one or one of," of,""exactly or "exactly one of." one of."

[0034] In the

[0034] the claims, In claims, as well as well the specification in specification as inasthe above, above, all transitional all transitional phrases suchsuch phrases as as "comprising," "including," "comprising," "including," "carrying," "carrying," "having," "having," "containing," "containing," "involving," "involving," "holding," "holding," "composedof," "composed of,"and andthethelike likeare aretotobebeunderstood understoodto to be be open-ended, open-ended, i.e.,totomean i.e., mean including including but but

not limited not limitedto. to.Only Onlythethe transitional transitional phrases phrases "consisting "consisting of and of and "consisting "consisting essentially essentially of of shall be shall be closed closed ororsemi-closed semi-closed transitional transitional phrases, phrases, respectively, respectively, as setin forth as set forth in the the United United States States Patent Patent

Office Manual ManualofofPatent PatentExamining Examining Procedures, Procedures, Section Section 2111.03. 2111.03.

[0035] As used

[0035] As used hereinherein in theinspecification the specification and inand the in the claims, claims, the phrase the phrase "at one," "at least least in one," in reference to reference to a list list ofofone one or or more elements, should more elements, should bebeunderstood understoodto tomean mean at least at least oneone element element

selected fromanyone selected from anyoneor or moremore of elements of the the elements in the in theof list list of elements, elements, but not but not necessarily necessarily

9 including including atatleast leastone oneof of each and and everyevery element specifically listed the within list the list of elements and 2023248067 10 Oct 2023 each element specifically listed within of elements and not excluding not excluding any anycombinations combinationsof of elements elements in the in the listofofelements. list elements.This This definition definition alsoallows also allows that elements that elementsmaymay optionally optionally be present be present other other than than the the elements elements specifically specifically identified identified within the within the list listof ofelements elements to to which the phrase which the phrase "at "at least least one" one" refers, refers, whether related or whether related or unrelated unrelated to to those those elements specifically elements specifically identified. identified. Thus, Thus, as as aa nonlimiting example, "at nonlimiting example, "at least least one one of of AAand andB"B"(or, (or, equivalently,"at"atleast equivalently, leastoneone of of A B," A or or or, B," equivalently or, equivalently "at one "at least least of one of AB") A and/or and/or B") can can refer, in refer, in one embodiment,to toatatleast one embodiment, leastone, one, optionally optionally including includingmore morethan than one,A,A,with one, with no no B present B present (and (and optionally including elements optionally including elements other otherthan thanB); B);ininanother anotherembodiment, embodiment, to least to at at least one, one, optionally optionally including more morethan thanone, one, B, B, with withnonoA Apresent present(and (andoptionally optionallyincluding includingelements elements other other than than A); A); in yet another embodiment, embodiment, totoatatleast least one, one, optionally optionally including includingmore morethan than one, one, A, A, andand at least at least one, optionallyincluding one, optionally including moremore than than one, Bone, (and B (and optionally optionally includingincluding other elements); other elements); etc. etc.

[0036] also also It should

[0036] It should be understood that, that, be understood in certain in certain methods methods described described herein that that include hereininclude morethan more than oneone stepstep or act, or act, the the order order ofsteps of the the steps or of or acts acts theof the method method is not necessarily is not necessarily limited tolimited to the order the order inin which whichthethe steps steps or or acts acts of the of the method method are recited are recited unlessunless the context the context indicates indicates

otherwise. otherwise.

The terms

[0037] The terms

[0037] "co-administration" "co-administration" and "co-administering" and "co-administering" or "combination therapy"therapy" or "combination refer refer to both to concurrent administration both concurrent administration (administration (administration ofoftwo twoorormore moretherapeutic therapeuticagents agentsat atthe thesame same time) and time) and time time varied varied administration administration (administration (administration of ofone oneorormore moretherapeutic therapeuticagents agentsatata atime time different fromthat different from that of of thethe administration administration of an of an additional additional therapeutic therapeutic agent or agent orasagents), agents), long as as long as

the therapeutic the therapeuticagents agents are are present present in patient in the the patient toextent, to some some preferably extent, preferably at amounts, at effective effective amounts, at at the the same time. In same time. In certain certain preferred preferred aspects, aspects, one one or more ofthe more of the present present compounds compounds described described

herein, are herein, arecoadministered coadministered in combination in combination with at with least at least one one additional additional bioactive bioactive agent, agent, especially especially including anananticancer anticanceragent. agent. In particularly In particularly preferred preferred aspects, aspects, the co-administration the co-administration of of compounds compounds results results in synergistic in synergistic activity activity and/orand/or therapy, therapy, including including anticancer anticancer activity. activity.

[0038] TheThe

[0038] termterm "compound", "compound", as used as used herein, herein, unless unless otherwise otherwise indicated,refers indicated, referstotoany any specific specific chemical compound chemical compound disclosed disclosed herein herein and and includes includes tautomers, tautomers, regioisomers, regioisomers, geometric geometric

isomers, and and where whereapplicable, applicable, stereoisomers, stereoisomers,including includingoptical optical isomers isomers(enantiomers) (enantiomers)andand other other

stereoisomers (diastereomers) stereoisomers (diastereomers)thereof, thereof, as well as well as pharmaceutically as pharmaceutically acceptable acceptable salts and salts and derivatives, including prodrug derivatives, prodrugand/or and/ordeuterated deuterated forms forms thereof thereof where where applicable, applicable, in context. in context.

Deuterated small Deuterated small molecules moleculescontemplated contemplated areare those those in in which which one one or more or more of the of the hydrogen hydrogen atomsatoms

contained in the contained in the drug drug molecule havebeen molecule have beenreplaced replacedbybydeuterium. deuterium.

10

[0039] Within

[0039] Within its use its use in context, in context, the the termterm compound compound generally generally refers refers to a single to a single compound, compound,

but also but also may may include include other other compounds compounds such assuch as stereoisomers, stereoisomers, regioisomers regioisomers and/or and/or optical optical isomers (includingracemic isomers (including racemic mixtures) mixtures) as well as well as specific as specific enantiomers enantiomers or enantiomerically or enantiomerically

enriched mixtures enriched mixtures ofofdisclosed disclosed compounds. compounds.TheThe termterm alsoalso refers, refers, in in context context to to prodrug prodrug forms forms of of compounds compounds which which have have beenbeen modified modified to facilitate to facilitate thethe administrationandand administration delivery delivery of of compounds compounds

to aa site to site of of activity. activity. It It is is noted notedthat thatinindescribing describing the the present present compounds, compounds, numerous numerous substituents substituents

and variables and variables associated associated with withsame, same,among among others, others, are are described. described. It understood It is is understood by those by those of of ordinary skill ordinary skill that that molecules molecules which whichareare described described herein herein are stable are stable compounds compounds as generally as generally

described hereunder. described hereunder. When the bond When the bondisisshown, shown,both botha double a double bond bond and and single single bondbond are are represented or represented or understood understoodwithin withinthe thecontext contextofofthe thecompound compound shown shown and well-known and well-known rules rules for for valenceinteractions. valence interactions.

[0040] The term

[0040] The term "Ubiquitin "Ubiquitin Ligase" refers refers Ligase" to a family to a family of proteins of proteins that facilitate the the that facilitate transfer transfer

of ubiquitin of ubiquitintotoa specific a specific substrate substrate protein, protein, targeting targeting the substrate the substrate protein protein for degradation. for degradation. For For example, cereblon example, cereblonisisananE3E3Ubiquitin Ubiquitin Ligase Ligase protein protein thatthat alone alone or combination or in in combination with with an E2 an E2 ubiquitin-conjugating enzyme ubiquitin-conjugating enzymecauses causes thethe attachment attachment of ubiquitin of ubiquitin to to a lysine a lysine on on a targetprotein, a target protein, and subsequently and subsequentlytargets targets the the specific specific protein protein substrates substratesfor fordegradation degradationby bythe theproteasome. proteasome. Thus, Thus,

E3 ubiquitin E3 ubiquitin ligase ligase alone alone or or in complex withananE2E2ubiquitin complex with ubiquitinconjugating conjugatingenzyme enzyme is responsible is responsible

for the transfer for the transferofofubiquitin ubiquitin to targeted to targeted proteins. proteins. In general, In general, the ubiquitin the ubiquitin ligase is ligase involvedis ininvolved in

polyubiquitination polyubiquitination suchsuch that that a second a second ubiquitin ubiquitin is attached is attached to the to the first; first; isa attached a third third is to attached the to the second, and second, andsosoforth. forth.Polyubiquitination Polyubiquitinationmarks marks proteins proteins for degradation for degradation by thebyproteasome. the proteasome. However,there However, thereare aresome someubiquitination ubiquitinationevents eventsthat thatare arelimited limitedtotomono-ubiquitination, mono-ubiquitination,ininwhich which only aasingle only singleubiquitin ubiquitinis isadded added by ubiquitin by the the ubiquitin ligaseligase to a substrate to a substrate molecule. molecule. Mono- Mono ubiquitinated proteins ubiquitinated proteins are are not not targeted targeted to to the the proteasome proteasomeforfordegradation, degradation, butbut maymay instead instead be be altered in altered in their their cellular cellularlocation locationororfunction, function,for forexample, example, via via binding other proteins binding other that have proteins that have

domainscapable domains capable of binding of binding ubiquitin. ubiquitin. Further Further complicating complicating matters,matters, differentdifferent lysines lysines on on ubiquitin can ubiquitin can be be targeted targeted by by ananE3E3to tomake make chains. chains. The The mostmost common common lysine lysine is Lys48is on Lys48 the on the ubiquitin chain. ubiquitin chain. This This isis the thelysine lysineused usedto tomake make polyubiquitin, polyubiquitin, whichwhich is recognized is recognized by the by the proteasome. proteasome.

[0041] term "patient" The "patient"

[0041] The term or "subject" or "subject" is used used throughout is throughout the specification the specification to describe to describe an an animal, preferably animal, preferably a human human ororaadomesticated domesticatedanimal, animal,totowhom whom treatment, treatment, including including prophylactic prophylactic

treatment,with treatment, withthethe compositions compositions according according to the present to the present disclosure disclosure is provided. is provided. Foroftreatment For treatment of

11 those infections, infections, conditions conditions or or disease states which are specific specific for for aa specific specific animal animal such as aa 2023248067 10 Oct 2023 those disease states which are such as humanpatient, human patient, the theterm termpatient patientrefers refers toto that that specific specific animal, animal, including including aa domesticated domesticatedanimal animal such such asasa adog dogor or catcat orfarm or a a farm animal animal such assuch as a cow, a horse, horse, cow,etc.sheep, sheep, etc. Iningeneral, In general, in the present the present disclosure, the term patient refers to a human patient unless otherwise stated or implied from the disclosure, the term patient refers to a human patient unless otherwise stated or implied from the context context ofofthe theuse useofof theterm. the term. The term

[0042] The term

[0042] "effective" "effective" is used is used to describe to describe an amount an amount of a compound, of a compound, composition composition or or component which, component which, when when usedused within within the context the context of intended of its its intended use, use, effects effects an intended an intended result. result.

The term The termeffective effective subsumes subsumesallallother othereffective effectiveamount amountor or effectiveconcentration effective concentrationterms, terms, which which

are otherwise described or used in the present application. are otherwise described or used in the present application.

Compounds andCompositions Compounds and Compositions

[0043] In In

[0043] oneone aspect,thethe aspect, descriptionprovides description compounds providescompounds comprising comprising an E3 E3 Ubiquitin an Ubiquitin Ligase binding Ligase binding moiety moiety("ULM") ("ULM")thatthat is is a cereblonE3E3 a cereblon Ubiquitin Ubiquitin Ligase Ligase binding binding moiety moiety ("CLM"). ("CLM").

In In one embodiment,the one embodiment, theCLM CLM is coupled is coupled to atochemical a chemical linker linker (L)(L) according according to the to the structure: structure:

(I) (I) L-CLM L-CLM wherein LLisis aa chemical wherein chemical linker linker group group and andCLM CLMis is a cereblon a cereblon E3 E3 Ubiquitin Ubiquitin Ligase Ligase binding binding moiety. moiety.

The number The number and/or and/or relative relative positions positions of the of the moieties moieties in compounds in the the compounds illustrated illustrated herein herein is is provided by provided byway wayofofexample example only. only. As As would would be understood be understood byskilled by the the skilled artisan, artisan, compounds compounds as as described herein can described herein canbebesynthesized synthesized with with any any desired desired number number and/orand/or relative relative position position of the of the

respective functional moieties. respective functional moieties.

[0044] TheThe

[0044] terms ULM ULM terms and CLM used are andareCLM their inclusive their ininclusive in used sense unless sense unless the context the context

indicates indicates otherwise. For example, otherwise. For example,the theterm termULMULM is inclusive is inclusive of all of all ULMs, ULMs, including including thosethose that that

bind cereblon bind cereblon (i.e., (i.e., CLMs). Further, CLMs). Further, thethe term term CLM CLM is inclusive is inclusive of allof all possible possible cereblon cereblon E3 E3 Ubiquitin Ligase binding Ubiquitin Ligase binding moieties. moieties. In another

[0045] In another

[0045] aspect, aspect, the present the present disclosure disclosure provides provides bifunctional bifunctional or multifunctional or multifunctional

PROTAC PROTAC compounds compounds useful useful for regulating for regulating protein protein activityactivity by inducing by inducing the degradation the degradation of a of a target protein. target In certain protein. In certain embodiments, thecompound embodiments, the compound comprises comprises a CLM acoupled, CLM coupled, e.g., e.g., linked linked covalently, directlyor or covalently, directly indirectly, indirectly, to atomoiety a moiety that abinds that binds targetaprotein target (i.e., protein (i.e., targeting protein protein targeting moiety or moiety or "PTM"). "PTM"). InIncertain certain embodiments, embodiments, the the CLM CLMand andPTMPTM are are joined joined or or coupledviaviaa coupled a chemical linker (L). chemical linker The CLM (L). The CLM recognizesthethecereblon recognizes cereblonE3E3ubiquitin ubiquitinligase ligase and and the the PTM PTM recognizes a atarget recognizes target protein proteinand andthethe interactionof of interaction thethe respective respective moieties moieties with with theirtheir targets targets

12 facilitates the degradation degradationof of the the target protein by placing the protein target in protein in proximity to the 2023248067 10 2023 facilitates the target protein by placing the target proximity to the ubiquitin ligase ubiquitin ligase protein. protein.An An exemplary bifunctional compound exemplary bifunctional compoundcancan be be depicted depicted as:as:

Oct (II) (II) PTM-CLM PTM-CLM

[0046] In In

[0046] embodiments,thethebifunctional certainembodiments, certain compound bifunctional compound further comprisesa achemical furthercomprises chemical linker linker ("L"). ("L"). For For example, the bifunctional example, the bifunctional compound compound cancan be be depicted depicted as:as:

(III) (III) PTM-L-CLM PTM-L-CLM wherein PTM wherein PTMis is a protein/polypeptidetargeting a protein/polypeptide targetingmoiety, moiety,L is L isa alinker, linker, and andCLM CLMis ais cereblon a cereblon E3 E3 ligase ligase binding binding moiety.

In certain

[0047] In certain

[0047] embodiments, embodiments, the compounds the compounds as described as described herein comprise multiple multiple herein comprise PTMs PTMs (targeting the same (targeting the sameorordifferent differentprotein proteintargets), targets),multiple multipleCLMs, CLMs, onemore one or or ULMs more(i.e., ULMs (i.e., moieties that moieties that bind bind specifically specifically to to another another E3E3Ubiquitin Ubiquitin Ligase, Ligase, e.g., e.g., VHL) VHL) or a or a combination combination

thereof. In thereof. In any any ofofthe the aspects aspects ofofembodiments embodiments described described herein, herein, the the PTMs, PTMs, CLMs,CLMs, and and ULMs ULMs can be coupled can be coupleddirectly directlyor or viavia oneone or more or more chemical chemical linkers linkers or a combination or a combination thereof. thereof. In In additional embodiments, additional where embodiments, where a compound a compound has multiple has multiple ULMs,ULMs, thecan the ULMs ULMs canthe be for be same for the same E3 UbiquintinLigase E3 Ubiquintin Ligaseororeach eachrespective respectiveULMULM can bind can bind specifically specifically to a to a different different E3 Ubiquitin E3 Ubiquitin

Ligase. In Ligase. In still still further furtherembodiments, whereaacompound embodiments, where compoundhashas multiple multiple PTMs, PTMs, the PTMs the PTMs can can bind bind the same the sametarget targetprotein protein or or each each respective respective PTM PTM can bindcan bind specifically specifically to a target to a different different target protein. protein.

[0048] In In

[0048] another another embodiment, the the embodiment, description compound providesa acompound descriptionprovides which which comprises comprises a a plurality ofofCLMs plurality coupled directly CLMs coupled directly or or via via aa chemical chemical linker linker moiety moiety (L). (L). For For example, example, aa compound having compound having twotwo CLMs CLMs can can be be depicted depicted as: as:

(IV) (IV) CLM-CLM CLM-CLM oror (V) (V) CLM-L-CLM CLM-L-CLM

[0049] In certain

[0049] where embodiments,where In certainembodiments, thecompound the compound comprises comprises multiple thethe CLMs, multipleCLMs, CLMs CLMs

are are identical. identical.InInadditional additionalembodiments, the compound embodiments, the compound comprising comprising a plurality a plurality of of CLMs CLMs further further

comprises at least comprises at least one PTMcoupled one PTM coupled to to a CLM a CLM directly directly or via or via a chemical a chemical linker linker (L)both. (L) or or both. In In

certain certain additional additionalembodiments, the compound embodiments, the compoundcomprising comprisinga plurality a pluralityof of CLMs CLMs further further

comprises multiplePTMs. comprises multiple PTMs.In In stilladditional still additional embodiments, embodiments,thethe PTMs PTMs are the are the samesame or, optionally, or, optionally,

different. In different. In still stillfurther embodiments, further embodiments,wherein wherein the the PTMs are different PTMs are different the respective respective PTMs may PTMs may

bindthe bind thesame same protein protein target target or bind or bind specifically specifically to a different to a different protein protein target.target.

13 embodiments, the description provides a compound comprising at least 2023248067 10 Oct 2023

[0050] In additional

[0050] In additional embodiments, the description provides a compound comprising at least

two different two different CLMs coupled CLMs coupled directly directly or or viavia a chemical a chemical linker linker (L)(L) or or both.ForFor both. example, example, such such a a compound having compound having twotwo different different CLMs CLMs candepicted can be be depicted as: as:

(VI) (VI) CLM-CLM' or CLM-CLM' or (VII) (VII) CLM-L-CLM' CLM-L-CLM' wherein CLM' wherein CLM' indicates indicates a cereblon a cereblon E3 Ubiquitin E3 Ubiquitin Ligase Ligase binding binding moiety moiety that that is structurally is structurally

different different from from CLM. CLM. In In certainembodiments, certain embodiments, the the compound compound may comprise may comprise a plurality a plurality of CLMsof CLMs

and/or and/or aa plurality plurality of CLM's.In In of CLM's. further further embodiments, embodiments, the compound the compound comprising comprising at least at twoleast two

different different CLMs, CLMs, a aplurality plurality of of CLMs, CLMs,and/or and/ora aplurality plurality ofofCLM's CLM's furthercomprises further comprises at leastoneone at least

PTMcoupled PTM coupled to CLM to a a CLM or a directly or a CLM' CLM' directly or via aorchemical via a chemical linker orlinker both. or In both. any of In theany of the embodiments described embodiments described herein,a compound herein, a compound comprising comprising at least at least two different two different CLMs CLMs can further can further

comprise multiple PTMs. comprise multiple PTMs.In In stilladditional still additional embodiments, embodiments,thethe PTMs PTMs are the are the samesame or, optionally, or, optionally,

different. In different. In still stillfurther embodiments, further embodiments,wherein wherein the the PTMs are different PTMs are different the the respective respective PTMs may PTMs may

bind the bind the same sameprotein proteintarget targetororbind bindspecifically specificallytotoa adifferent differentprotein proteintarget. target. InInstill still further further embodiments, thePTM embodiments, the PTM itselfisisaaULM itself ULMor or CLMCLM (or ULM' (or ULM' or CLM'). or CLM').

[0051] In In

[0051] a preferredembodiment, a preferred CLMCLM thethe embodiment, comprises comprises a moiety that that a moiety of of a ligand is aisligand thethe

cereblon E3 Ubiquitin cereblon E3 Ubiquitin Ligase Ligase (CRBN). (CRBN).In certain In certain embodiments, embodiments, the comprises the CLM CLM comprises a a chemotype from chemotype from the"imide" the "imide" classofofofofmolecules. class molecules.In In certainadditional certain additionalembodiments, embodiments, the the CLM CLM

comprises comprises aa phthalimido phthalimido group group ororanananalog analogor or derivativethereof. derivative thereof. In still In still additional additional

embodiments, theCLMCLM embodiments, the comprises comprises a phthalimido-glutarimide a phthalimido-glutarimide group group or or an oranalog an analog or derivative derivative

thereof. In still thereof. In still other other embodiments, theCLM embodiments, the CLM comprises comprises a member a member of the of the consisting group group consisting of of thalidomide, lenalidomide, thalidomide, lenalidomide, pomalidomide, pomalidomide,andand analogs analogs or or derivativesthereof. derivatives thereof.

[0052] In In

[0052] embodiments,thethe additionalembodiments, additional description provides description the compounds provides the described compoundsas asdescribed herein including herein including their their enantiomers, enantiomers, diastereomers, diastereomers, solvates solvates and and polymorphs, polymorphs,including including pharmaceutically acceptable salt forms thereof, e.g., acid and base salt forms. pharmaceutically acceptable salt forms thereof, e.g., acid and base salt forms.

Exemplary

[0053] Exemplary

[0053] Cereblon Cereblon Binding Binding and/or and/or Inhibiting Inhibiting Compounds Compounds

[0054] In one

[0054] In one aspect aspect the description the description provides provides compounds compounds usefuluseful for binding for binding and/orand/or inhibiting inhibiting

cereblon E3Ubiquitin cereblon E3 UbiquitinLigase Ligase binding binding moiety. moiety. In certain In certain embodiments, embodiments, the compound the compound has a has a chemical structure that chemical structure that includes includes atat least least one one ofof(e.g., (e.g., the the compound compoundhas has a chemical a chemical structure structure

selected fromthethe selected from group group consisting consisting of): of):

14

Oct 2023

[0055]

[0055] Neo-imide Compounds Neo-imide Compounds

[0056]

[0056] In In one aspect the one aspect the description description provides provides compounds usefulfor compounds useful forbinding bindingand/or and/orinhibiting inhibiting cereblon. In certain cereblon. In certain embodiments, thecompound embodiments, the compound is selected is selected from from thethe group group consisting consisting of of

chemical structures: chemical structures: 2023248067 10

X X G X X G Q4 on3 Q 03 N Q Q3 Q N Q N A =ZZ N Norm N N> Z

Q W A Q W N Q Rn Q Rn Rn Rn ZRn Rn R R' G' Z

(al) (al) (b) (b)

G G x X .1 N z Z X X X G xX X G Q N Q4 N 03 Q Q N oa N Normal Z 1/ N Z Rn Q2 Rn~ Q W w A A N NQI Q Y,, Y Z Z Rn Rn X X G' G' Rn Rn Q (c) (c) (dl) (d1)

G G

X x N1 z Z x X x X x X

Q4 Q4 3Q4 X NR4 oa Q of Q3 N Rn Norm Z

Q Q W kr N N A A RnNvwR Q Rn Rn Q Rn Rn Rn Rn

(e) (e) (f) (f)

G 2023 G

X N Z

. X X X 2023248067 10 Oct X X G Q4 X/ Q4 Q4 N of Q; W3 Q33 N N W N Norm Z Z Rn Rn Q2 Q2 W A Rn/Rn Q Rn Rn Q Rn Rn

(a2) (a2) (d2) (d2)

X XN/GG

Q4 N 03 Q N Z /

W4 Norm Z Q2 Q1 W A Q W A Rn Rn Rn Rn

(a3) (a3) wherein: wherein:

W W ofofFormulas Formulas(a)(a) through through (e)(e) is is independently independently selected selected fromfrom the the group group CH2 C=O, CH, CHR, , CHR, C=O, NH,cyclopropyl S02,NH, SO, cyclopropyl group, group, cyclobutyl cyclobutyl group, and and group, N-alkyl; N-alkyl;

W isisselected W3 fromC CororN;N; selectedfrom each each XX of ofFormulas Formulas(a) (a)through through(e) (e)isis absent absent or or independently independentlyselected selected from fromthe thegroup groupO 0 andand

S; S;

Y of Y of Formulas Formulas(a) (a)through through(e)(e)isisindependently independentlyselected selectedfrom from thethe group group CH, CH 2 , -C=CR', -C=CR', NH, NH, N-alkyl, N-aryl, N-alkyl, N-aryl, N-hetaryl, N-hetaryl, N-cycloalkyl, N-cycloalkyl, N-heterocyclyl, 0, and N-heterocyclyl, O, and S; S; each each ZZ of of Formulas Formulas(a) (a) through through(e) (e) isis absent absent or or independently independently selected selected from fromthe thegroup groupO 0andand S, except S, except that that both both XX and and Z cannot cannot be be absent; absent; G andG'of G and G'of Formulas Formulas (a) (a) through through (e) independently (e) are are independently selected selected from from the the H, group group H, alkyl alkyl

(linear, (linear,branched, optionally branched, substituted), optionally OH, R'OCOOR, substituted), R'OCONRR", OH, R'OCOOR, R'OCONRR", CH CH-2 heterocyclyloptionally heterocyclyl optionally substituted substituted with with R',benzyl R', and and benzyl optionally optionally substituted substituted with R'; with R'; Q1 -- Q4 Q1 Q4ofofFormulas Formulas (a)(a)through through(e)(e)represent representa carbon a carbon C substitutedwith C substituted witha group a group independently selected from independently selected fromR', R', NNororN-oxide; N-oxide; A ofFormulas A of Formulas(a)(a) through through (e) (e) is independently is independently selected selected from from the group the group H, (linear, H, alkyl alkyl (linear, branched,optionally branched, optionally substituted), substituted), cycloalkyl, cycloalkyl, Cl andCl F;and F;

16

2023 R of R of Formulas Formulas(a) (a) through through(e) (e)comprises, comprises,but butisis not not limited limited to: to: -CONR'R", -CONR'R", -OR', -OR', -NR'R", -NR'R",

SR', -SOR', SR', -SO2NR'R", -SO2R', -SONR'R", -CR'R"-, -CR'R"-, -CR'NR'R"-, -CR'NR'R"-, (-CR'O),,R", (-CR'O)'R", -aryl,-aryl, -hetaryl, -hetaryl, -alkyl -alkyl 2023248067 10 Oct

(linear, (linear, branched, optionally substituted), branched, optionally substituted), -cycloalkyl, -cycloalkyl, -heterocyclyl, -heterocyclyl, -P(O)(OR')R", -P(O)(OR')R",-

P(O)R'R", -OP(O)(OR')R", P(O)R'R", -OP(O)(OR')R", -OP(O)R'R", -OP(O)R'R",-Cl, -Cl, -F, -F, -Br, -Br,-I, -CF-CF, -I, 3 , -CN, -CN,-NR'SO 2NR'R", -NR'SONR'R",-

NR'CONR'R",-CONR'COR", NR'CONR'R", -CONR'COR", -NR'C(=N-CN)NR'R", -NR'C(=N-CN)NR'R", -C(=N-CN)NR'R", -C(=N-CN)NR'R", -NR'C(=N -NR'C(=N- CN)R", -NR'C(=C-NO2)NR'R", CN)R", -NR'C(=C-NO)NR'R", -SO2NR'COR", -SO2NR'COR", -NO,-NO2 , -CO -COR', 2 R', -C(C=N-OR')R", -C(C=N-OR')R", - CR'=CR'R", -CCR',-S(C=O)(C=N-R')R", CR'=CR'R", -CCR', -S(C=O)(C=N-R')R", -SFand -SF5 5 and -OCF 3 -OCF

R' andR"R"ofofFormulas R' and Formulas (a) (a) through through (e) are (e) are independently independently selected selected from from a bond, a bond, H, alkyl, H, alkyl,

cycloalkyl, aryl, heteroaryl, cycloalkyl, aryl, heteroaryl, heterocyclic, heterocyclic, -C(=O)R, -C(=)R, heterocyclyl, heterocyclyl, eacheach of which of which is is optionally substituted; optionally substituted;

n' of n' of Formulas Formulas (a)(a) through through (e) (e) is integer is an an integer from from 1-10 (e.g., 1-10 (e.g., 1-4); 1-4); - ~~~~ of Formulas of Formulas(a) (a)through through(e) (e) represents represents aa bond bondthat that may maybebestereospecific stereospecific ((R) ((R)oror(S)) (S)) or or non-stereospecific; non-stereospecific;

' represents represents aa single single bond bond or or aa double double bond; bond;

representsa abond represents bond that that maymay be stereospecific be stereospecific ((R) or((R) (S))or or(S)) or non-stereospecific; non-stereospecific; and and Rn comprises1-41-4independent Rn comprises independent functional functional groups, groups, optionally optionally substituted substituted linear linear or branched or branched

alkyl (e.g., aa C1-C6 alkyl (e.g., linear oror branched C1-C6 linear branchedalkyl alkyloptionally optionallysubstituted substitutedwith with one one or more or more

halogen, cycloalkyl halogen, cycloalkyl (e.g., (e.g., aa C3-C6 C3-C6cycloalkyl), cycloalkyl),or or aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), optionally optionally

substituted aryl(e.g., substituted aryl (e.g.,ananoptionally optionally substituted substituted C5-C7C5-C7 aryl), optionally aryl), optionally substituted substituted alkyl- alkyl aryl (e.g., an aryl (e.g., an alkyl-aryl alkyl-arylcomprising comprising at least at least one one of an of an optionally optionally substituted substituted C1-C6 alkyl, C1-C6 alkyl,

an optionally substituted an optionally substituted C5-C7 C5-C7aryl, aryl,or or combinations combinations thereof), thereof), optionally optionally substituted substituted

alkoxyl group alkoxyl group(e.g., (e.g., aa methoxy, methoxy,ethoxy, ethoxy,butoxy, butoxy, propoxy, propoxy, pentoxy, pentoxy, or hexoxy; or hexoxy; wherein wherein

the alkoxyl the alkoxyl may maybebesubstituted substitutedwith with oneone or or more more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl,

cycloalkyl (e.g.,a aC3-C6 cycloalkyl (e.g., C3-C6 cycloalkyl), cycloalkyl), or arylor(e.g., aryl C5-C7 (e.g., aryl)), C5-C7optionally aryl)), optionally substituted substituted

X z Z y (e.g., optionally (e.g., substituted with optionally substituted with one oneor or more more

halogen,alkyl, halogen, alkyl,haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., (e.g., a C3-C6a cycloalkyl), C3-C6 cycloalkyl), or aryl (e.g., or aryl (e.g.,

0 O x C5-C7 aryl)), optionally C5-C7 aryl)), optionallysubstituted substituted Yy (e.g., optionally (e.g., optionally

17 substituted withoneone or more halogen, alkyl, alkyl, haloalky, fluoroalkyl, cycloalkyl (e.g., a C3 2023248067 10 Oct 2023 substituted with or more halogen, haloalky, fluoroalkyl, cycloalkyl (e.g., a C3-

C6 cycloalkyl),or or C6 cycloalkyl), aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), or atoms; or atoms; and and each ofX,x,y,y, and each of andZ zare areindependently 0, 1,0,2,1,3,2,4,3,5, independently 4,or5,6,or 6,

[0057]

[0057] Exemplary CLMs Exemplary CLMs

[0058]

[0058] In In any of the any of compounds the compounds described described herein, herein, CLMCLM thethe comprises comprises a chemical a chemical structure structure

selected selected from the group: from the

X X G X X G N/XG X X G Q4 Q4 N N on Q Q3 Norm Z N Z Q2 Q W A W N Q Rn Q n Rn Rn ZRnNRZ Rn R' G' ,

(a1) (al) (b) (b)

G A "/ I Y Z G X N Z X X G N X N

Q4 Q4 N 03 Q4 Q N ZQ4 03 Q N Novr Z Rn

Q WN W A N N ZZ Q Rn

Rn Q X xG' z rJJRn Rn Q Y Z Rn X x xx x (c) (c) (dl) (d1)

G G N' A N Q NY.n, QX X NN Z4 Z X X X Q4 Q4 Q N N Q3 O3 Rn Norrn Z

Q Q W Rn Q N A Rn Q Rn Rn Rn Rn

(e) (e) (f) (f)

18

Oct 2023 G G

X N Z

. X X X X X G Q4 X/ Q4 Q4 N W3 Q; Q3 N N 2023248067 10 & W N Norm Z Z & Rn Rn Q2 Q2 W A Rn/Rn Q Rn Rn Q Rn Rn

(a2) (a2) (d2) (d2)

X XN/GG

Q4 N 03 N Z /

W4 Normal Z Q2 Q1 W A Q W A Rn Rn Rn Rn

(a3) (a3) wherein: wherein:

W W ofofFormulas Formulas(a)(a) through through (f) (f) is is independently independently selected selected from from the the group group CH2 C=O, CH, CHR, , CHR, C=O, NH, S02,NH, SO, N, N, optionally optionally substituted substituted cyclopropyl cyclopropyl group, group, optionally optionally substituted substituted cyclobutyl cyclobutyl

group, and N-alkyl; group, and N-alkyl; W isisselected W3 fromC CororN;N; selectedfrom each each XX of ofFormulas Formulas(a) (a)through through(f) (f) isis absent absent or or independently independently selected selected from fromthe thegroup groupO 0andand S; S;

Y of Y of Formulas Formulas(a) (a)through through(f)(f)isis independently independentlyselected selectedfrom from thethe group group CH2, CH2, -C=CR', -C=CR', NH, NH, N-alkyl, N-aryl, N-alkyl, N-aryl, N-hetaryl, N-hetaryl, N-cycloalkyl, N-cycloalkyl, N-heterocyclyl, 0, and N-heterocyclyl, O, and S; S; each each ZZ of of Formulas Formulas(a) (a)through through(f) (f) is is absent absent or or independently independently selected selected from fromthe thegroup groupO 0 andand

S, except S, except that that both both X X and and Z cannot be Z cannot be absent; absent; G andG'G'ofof G and Formulas Formulas (a) (a) through through (f) are (f) are independently independently selected selected from from the group the group H, alkyl H, alkyl

(linear, (linear,branched), OH, branched), OH, R'OCOOR, R'OCONRR", R'OCOOR, R'OCONRR", CH2-heterocyclyl CH-heterocyclyl optionally optionally substituted withR',R',andand substituted with benzyl benzyl optionally optionally substituted substituted with R'; with R';

Q1 -- Q4Q4 Q1 of of Formulas Formulas (a)(a) through through (f)(f)represent representa acarbon carbonC substituted C substitutedwith witha agroup group independently selected from independently selected fromR', R', NNororN-oxide; N-oxide; A ofofFormulas A Formulas(a)(a) through through (f) (f) is is independently independently selected selected fromfrom the group the group H, (linear, H, alkyl alkyl (linear, branched,optionally branched, optionally substituted), substituted), cycloalkyl, cycloalkyl, Cl andCl F;and F;

19

Oct 2023 R of R of Formulas Formulas(a) (a) through through(f) (f) comprises, comprises,but butisis not not limited limited to: to: -CONR'R", -CONR'R", -OR', -OR', -NR'R", -NR'R",-

SR', -SO2R', SR', -SO2R',-SO2NR'R", -SO2NR'R", -CR'R"-, -CR'R"-, -CR'NR'R"-, -CR'NR'R"-, (-CR'O),,R", (-CR'O)'R", -aryl, -alkyl -aryl, -hetaryl, -hetaryl, -alkyl (linear, (linear, branched, optionally substituted), branched, optionally substituted), -cycloalkyl, -cycloalkyl, -heterocyclyl, -heterocyclyl, -P(O)(OR')R", -P(O)(OR')R",-

P(O)R'R",-OP(O)(OR')R", P(O)R'R", -OP(O)(OR')R", -OP(O)R'R", -OP(O)R'R", -Cl,-Br, -Cl, -F, -F, -I, -Br, -CF3, -I, -CF3, -CN,-CN, -NR'SO2NR'R", -NR'SO2NR'R",- 2023248067 10

NR'CONR'R",-CONR'COR", NR'CONR'R", -CONR'COR", -NR'C(=N-CN)NR'R", -NR'C(=N-CN)NR'R", -C(=N-CN)NR'R", -C(=N-CN)NR'R", -NR'C(=N -NR'C(=N- CN)R", -NR'C(=C-N02)NR'R",-SO2NR'COR", CN)R", -NR'C(=C-NO2)NR'R", -SO2NR'COR", -N02, -NO2, -CO2R', -CO2R', -C(C=N-OR')R", -C(C=N-OR')R", - CR'=CR'R", -CCR',-S(C=O)(C=N-R')R", CR'=CR'R", -CCR', -S(C=O)(C=N-R')R", -SF5 -SF5 andand -OCF3 -OCF3

R' andR"R"of of R' and Formulas Formulas (a) (a) through through (f) are (f) are independently independently selected selected from from a bond, a bond, H, alkyl, H, alkyl,

cycloalkyl, aryl, heteroaryl, cycloalkyl, aryl, heteroaryl, heterocyclic, heterocyclic, -C(=O)R, -C(=)R, heterocyclyl, heterocyclyl, eacheach of which of which is is optionally substituted; optionally substituted;

n'of n' of Formulas Formulas (a)(a) through through (f) (f) is integer is an an integer from from 1-10 (e.g., 1-10 (e.g., 1-4); 1-4);

' represents represents aa single single bond bond or or aa double double bond; bond;

of Formulas of Formulas (a)(a) through through (f) represents (f) represents a bonda that bondmaythat may be stereospecific be stereospecific ((R) ((R) or (S)) or or (S)) or non-stereospecific; non-stereospecific;

Rn comprises1-41-4independent Rn comprises independent functional functional groups, groups, optionally optionally substituted substituted linear linear or branched or branched

alkyl (e.g., aa C1-C6 alkyl (e.g., linear oror branched C1-C6 linear branchedalkyl alkyloptionally optionallysubstituted substitutedwith with one one or more or more

halogen, cycloalkyl halogen, cycloalkyl (e.g., (e.g., aa C3-C6 C3-C6cycloalkyl), cycloalkyl),or or aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), optionally optionally

substituted aryl(e.g., substituted aryl (e.g.,ananoptionally optionally substituted substituted C5-C7C5-C7 aryl), optionally aryl), optionally substituted substituted alkyl- alkyl aryl (e.g., an aryl (e.g., an alkyl-aryl alkyl-arylcomprising comprising at least at least one one of an of an optionally optionally substituted substituted C1-C6 alkyl, C1-C6 alkyl,

an optionally optionally substituted substituted C5-C7 C5-C7aryl, aryl,or or combinations combinations thereof), thereof), optionally optionally substituted substituted

alkoxyl group alkoxyl group(e.g., (e.g., aa methoxy, methoxy,ethoxy, ethoxy,butoxy, butoxy, propoxy, propoxy, pentoxy, pentoxy, or hexoxy; or hexoxy; wherein wherein

the alkoxyl alkoxyl may maybebesubstituted substitutedwith with oneone or or more more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl,

cycloalkyl (e.g., (e.g., aa C3-C6 cycloalkyl), or C3-C6 cycloalkyl), or aryl aryl (e.g., (e.g., C5-C7 aryl)), optionally C5-C7 aryl)), optionally substituted substituted

X z Z y (e.g., optionally (e.g., substituted with optionally substituted with one oneor or moremore

halogen,alkyl, halogen, alkyl,haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., (e.g., a C3-C6a cycloalkyl), C3-C6 cycloalkyl), or aryl or aryl (e.g., (e.g.,

O' 0

X C5-C7 aryl)), optionally C5-C7 aryl)), optionallysubstituted substituted y (e.g., optionally (e.g., optionally

20

Oct 2023 substituted withoneone substituted with or more or more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., a C3 (e.g., a C3-

C6 cycloalkyl),or or C6 cycloalkyl), aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), or atoms; or atoms; and and each ofx,x,y,y, and each of andZ zare areindependently independently 4,or5,6.or 6. 0, 1,0,2,1,3,2,4,3,5,

[0059] any aspect In aspect

[0059] In any or embodiment or embodiment described herein,herein, described the of each ofeach the ZX of X and and of the 6-member theZ 6-member 2023248067 10

monocycliccyloalkyl monocyclic cyloalkylorormonocyclic monocyclic heterocycloalkyl heterocycloalkyl of of thethe CLMCLM are each are each independently independently absent, absent,

o or S,S, except O or exceptthat thatboth bothX X andand Z cannot Z cannot be absent. be absent. In anyInaspect any aspect or embodiment or embodiment describeddescribed

herein, X herein, on the X on the middle middle ring ring is is selected selected from from O0and andS,S, and andeach eachofofthe the XXand andZ Zofofthe the6-member 6-member monocycliccyloalkyl monocyclic cyloalkylorormonocyclic monocyclic heterocycloalkyl heterocycloalkyl of of thethe CLMCLM are each are each independently independently absent, absent,

oO or or S, S, except that that both both X X and Z Z cannot cannot be be absent. absent.

[0060] In In

[0060] certain embodiments certainembodiments described the CLM herein, the describedherein, CLM or or ULM ULM comprises comprises a chemical a chemical

structure selectedfrom structure selected from thethe group: group:

0 O 0 O NH NH N 0 O WR W R Rn Rn Formula(g) Formula (g) wherein: wherein:

WofofFormula W Formula(g) (g)isis independently independentlyselected selectedfrom fromthe thegroup groupCH, CH 2 , C=0, C=0, NH,N-alkyl; NH, and and N-alkyl; R of R of Formula Formula(g)(g)isisindependently independentlyselected selectedfrom from a H, a H, methyl, methyl, alkyl alkyl (e.g.,a or (e.g., a or C1-C6 C1-C6 alkyl alkyl

(linear, (linear, branched, optionally branched, optionally substituted)); substituted);

- of Formula of Formula(g)(g)represents representsa bond a bond thatthat may may be stereospecific be stereospecific ((R) ((R) or (S))oror(S)) non-or non stereospecific; and stereospecific; and

Rn comprises1-41-4independent Rn comprises independent functional functional groups, groups, optionally optionally substituted substituted linear linear or branched or branched

alkyl (e.g., aa C1-C6 alkyl (e.g., linear oror branched C1-C6 linear branchedalkyl alkyloptionally optionally substituted substituted with with one one or more or more

halogen, cycloalkyl halogen, cycloalkyl (e.g., (e.g., aa C3-C6 C3-C6cycloalkyl), cycloalkyl),or or aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), optionally optionally

substitutedaryl substituted aryl(e.g., (e.g.,ananoptionally optionally substituted substituted C5-C7C5-C7 aryl), optionally aryl), optionally substituted substituted alkyl- alkyl aryl (e.g., aryl (e.g., an an alkyl-aryl alkyl-arylcomprising comprising at least at least one one of an of an optionally optionally substituted substituted C1-C6 alkyl, C1-C6 alkyl, an optionally substituted an optionally substituted C5-C7 C5-C7 aryl,or or aryl, combinations combinations thereof), thereof), optionally optionally substituted substituted

alkoxyl group alkoxyl group(e.g., (e.g., aa methoxy, methoxy,ethoxy, ethoxy,butoxy, butoxy, propoxy, propoxy, pentoxy, pentoxy, or hexoxy; or hexoxy; wherein wherein

21

Oct 2023 the alkoxyl alkoxyl may maybebesubstituted substitutedwith with oneone or or more more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl,

cycloalkyl (e.g., aa C3-C6 cycloalkyl (e.g., cycloalkyl), or C3-C6 cycloalkyl), or aryl aryl (e.g., (e.g., C5-C7 aryl)), optionally C5-C7 aryl)), optionally substituted substituted

offx X 2023248067 10 z Z y (e.g., optionally (e.g., substituted with optionally substituted with one oneor or more more

halogen,alkyl, halogen, alkyl,haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., (e.g., a C3-C6a cycloalkyl), C3-C6 cycloalkyl), or aryl or aryl (e.g., (e.g.,

O' 0

[0061] C5-C7 aryl)), optionally C5-C7 aryl)),

substituted substituted with

C6 optionallysubstituted withoneone cycloalkyl),or or C6 cycloalkyl),

[00611 In any In of or more

aryl aryl

anytheofembodiments substituted or more halogen, halogen,

(e.g.,C5-C7 (e.g.,

the embodiments C5-C7

described described Xof alkyl, alkyl,

aryl)), aryl)), haloalky, haloalky,

or atoms. or atoms.

herein, herein, the W, the W,Z,X,G,Y,G', X, Y, y fluoroalkyl, fluoroalkyl,

Z, R, G, R', G',R", (e.g., optionally (e.g.,

cycloalkyl cycloalkyl

R, R', Q1- optionally (e.g., a C3 (e.g., a C3-

R", QI Q4, A, and Q4, A, andRnRnofofFormulas Formulas (a)(a) through through (g)(g) cancan independently independently be covalently be covalently coupled coupled to a linker to a linker

and/or a linker and/or a linker to towhich which is isattached attachedone oneor ormore more PTM, ULM, PTM, ULM, CLMCLM or CLM' or CLM' groups.groups.

[0062] More

[0062] More specifically, non-limiting examples specifically,non-limiting CLMs of CLMs examples of includethose include shown thoseshown below below as as well as as those those "hybrid" "hybrid" molecules moleculesthat thatarise arisefrom fromthe thecombination combination ofor1 more of 1 or more of the of the different different

features features shown in the shown in the molecules below. molecules below.

22

0 0 2023248067 10 Oct 2023

0 o 0 O 0 o o0 o O NH HN HN HN NH O 0 o N o pNH 0 4Null""I 0 =

Rn Rn 4N

0 O Rn Rn 0Rn o - Rn 0

0 o Ss 0 o O0 0 O o0 NH HN HN HN

N 0 NNH NI11111- 0 o NN o0 4NH O MIIN Ali Alk Alk Alk

Rn Rh 0 Rn Rn oRn Rh o0 o O

Oo 0 0 O 0 o 0 o O0 o

N,,NH NNNH HN N~NH HN HN 0 N _ 0 N N _ S N N _ o N O

Rn Rn Rn Rn 0 o Rn S

0 0 O 0 O o O o0 o0 o N NH NN N HN N NNH HN HN N N o N o N o 0 4N 0 N 0N

Rn N 0 Rn 0n Rnxo Rn o R o R o

0 o0 o0 0 0 o0 o o o N N N HN HN HN N 0H N 0H N N 0H N o N o N o N c"N N, L N 0 Rn 0Rn 0 Rn Rh Rh Rn o o 00 / 0 O 0 o 0 o o o O NHN,,NH HN N,,N N/,,, HN N - =0 N N _ N N - o0 N o

Rn Rn 0 Rn Rn O 0 Rn RE o0 o

0 o 0 OH 0 o 0 o 0 o o0 o HO

N,,NNN ~NHHN ', N

0 N 0 NN 0 o N N o N o OS S

0 Rn Rnx Rn Rn Rn R o

23

0 0 0 0 O 0 o 0 o o o o NH NH NH NH NH NH N S N O N o N NH 0 N NH N NH NH NH NH RnY-Rn Rn Rn s S Rnp Rn 0 O

0 0 0 o0 o 0 o O o o o - NH N zN H NNH N" NH N NH NH N -0 N N0N N o N o N >= o NH 0 L NH NH N N NH NH NH Rn 0 0 Rn Rn RnRn Rn O Rn o o

o 0 0 0 0 O o0 o o o o

N4 N - N 0 NN4 N N NH N NH NH NH0N NH0 N N o N o N O NNY- 0 N N NH N NH NH NH N NH Rn Rn Rn Rn 0 Rn0 Rn o O o

o OH O o o o 0 o 0 NzOH 0 0 N N 0 0 N/ N NH N NHo> N N S N O N o N NH 0 \ NH 0NH NH NH NH Rn Rn Rn Rn s S Rn0 Rn o

o o o o o o N NH NH NzNHNH N

0 N 0= N N S NN o N o NH NH NH OS S SNHNH NH

Rny Rn O2Rn Rn 0Rn o Rn 0 o

24 o 0 0 0 0 o 0 O O O o NH HN NH NH N N0 HN HN >==o" 0- N > o NN 0 O 0011111N O NH N NH HN HN HN RnN nH Rn Rn 0 S S Rn Rn O O O o O0 Rn0 0 Rn0 o0 s o O 0 O 0 o 0 O S

NH HN NH HN NNH HN NI~I'0 N N 0 o NN o0 millin o

E Alk Ali NH HN Alk RnAlk HN NH HN NHp R0 Rn 0 Rn Rn 0 0 Rn 0 o O o o 0 O

0 o0 0 0 O 0 o O0 O O

NH NH NH HN N HN HN N- 0= N- N > s S NN- o0= N o NH NH HN NH HN HN 0 Rn -4 0 Rnp 0- 0 Rn Rin Rn o Rn o S o O o 0

o 0 0 o 00 o o O0 o o NN N HN N HN HN N czN- 0= o o0 N NN o0 NN

NH NN N HN NH HN HN Rn 0 O O0 Rn Rn 0 O 0 0 Rn Rn 0 o 0 o R

Oo O0 o0 o0 o0 o0

NN NH HN NNN N HN ' N ~NHHN N NN- 0=o r NN o01 N- N 0 o o N HN HN HN N NH Rn 0N0 0 0- N Rn Rn Rn Rn o Rn o o o O o 0 0 0 0 O 00

o O o o o Ho o N/0 0 N /O 0 NH 0 0 N N HN NN 0 o N o NN 0o O 4N 0 NH NH HN NH HN HN Rn 0 0 Rn Rn o0 0 o Rn Rn 0Rn O O o

0 0 O 0 o 0 o o00 O O NH NH N HN HN

N o0 NN 0 o 0 o NH S NH HN HN HN °0 Rn 0- 0 H Rn Rn 02 0 o Rn Rn0H O o o R o

25

H HN H HN H ZI 2023 O, 0 OO 0 0 N O 0 0s 'N N o o O S

MIIIIIIII

2023248067 10 Oct N N N N'~

O O0 o 0 o Rn Rn HNN Rn Rn HHN Rn Rn HIZ 0 o N 00 N 0S o S N 0 o o o 0 0 o 0 o O

All

N NNN" N-. N N N Alk Alk Alk Alk

0 o 0 o 0 o IZ IZ Rn HNN RnH RnH Rn oO o 0 Rn N o 0 o Rn Rn 0 O N S S o o 0 o 0 o

NN NN N N

S S 0 o Rn NN HN Rn Rn Rn Rn Rn 0 O, 0H IZ H HN

o o O O o 0 0 o O

N 'N NN NPN N N "N N- N N N N 0 o K L Rn H00 ZI o O Rn 0 N 0 Rn H Rn H o o Rn ZI Rn ZI

O 0 o 0 0NO O 0 N o o N N N NN N N N. N

NN 0 o N, N .

RnyN Rn N 0 o 0 O Rn Rn Rn Rn H HN

0 N 0H IZ H O O HN

o o o O O 0 0 o o N N NN N N N N-N NNN IZ N H O 0N0 Rn Rn O N O Rn Rn Rn OH Rn OH H HN

0 OO N N O 0 0 o 0 N O 00 O NJ o 0 o 0 o

" N N N N N " N

0 S 0~ 0 o S O Rn o Rn Rn Rn Rn Rn

26

H IZ H IZ

N 0 o O0 N O o 0 o O N 0 O 0 N

N NN: N Nr N

O 0 O0 o Rn Rn Rn Rn Rn Rh

H IZ

O o O

0 N 0 0 o Rh IZ

O o 0 0 o o o o 0 N 0 N N N 00 o N N o0

N / / Rn Rn HN Rn Rn Rn Rn 0 0 o 0 O O

o R0 O 0 O o 'o 0 o Rn Rn N N 0 O Rn0N O NH HN

NH NH NNt =0 o HN HN 0 0 Rn O O Rn o 0 o0 0 o O o NH NH HN HN NH 0= NNt H= N o O

Rn Rn Rn Rn o O o o

27 LT

F E

O o NNH F O HN F 0 00 0 o N N 0= O NH HN

N O

Rn O Rnn o Rn o O O0 0 O 0 o

HN NH , HN NH N N O N O N

Rn Rü Rn Rn o O 0 o NNNH O O HN 0N0 HN N o N O

Rn Rn O0 o

o 0 0 0, o O o o

HN HN N 0 NN 0 N o N o

Rn 0 Rn 0 Rü o Rn o o O 0 o 0 O 0 0 HN HN N N0 N 0 0H Nt N o

Rn Ri 0 Rn 0 O o R

28

H IZ H 0 O N 0 O 0 o O0 0 o NH HN NN N N NNt N 0 =o

Rn Rn Rn Rn 0 X O

0 O O0 x

HN NH N HN N N NN- = O N o NCH= N Rn Rn O0 Rn Rn H ZI

0 o N 0 O 0 O

N N

I cxNN Rn Rn

29

O o 0 o o 0 O 00 o O 0

NHNHN HN HN HN

N 0 millin N0l" o0 N o O

RnzRn Rn Rn Rn R 0 o S o0 0 O O o0 0 s

NH HN NH HN NH HN N 0 NI11,i"- millin 0 N N - o 0 N Fill O o

Alk Alk Alk Alk Rn Rin Rn Rh Rn Rn

oo 0 o S 0 o o 0 o 0

NH NH HN ',NH HN HN NNHN0 N N S N o o

RnpRn Rin Rn Rn R o0 o oo 0 o 0 o 0 O 0

N N HN N NH) NH HN NH HN N N- 0 N4 N N o N 0N0 o N o N ll&Rn Rin Rn RnpRn Rin 0 o O 0 OH 0 0 o o HO 0 0 o o o o N N NH NNH N N N HN N~N HN N N o N o N O N

Rh o Rn N N -0 R N- 0N N 0

o o O o o

Rn 0RnR HN N HN

N N N o o 0~~~ 0 F00 Rn Rin Rn

0 0 NH

o o o O Rn Rn N HN N o N O 20s

Rn Ri

300

S S 0 0 o 0 o o 0 o 0 o OH OH / NH NH N NH N S N NNN N NH o N o NH NH NH Rn Rn 0 N Rn N NH NH NNH NHN

o o O o o o Rn Rn 2023248067 Rn R N NH N NH NH N N O N o N o 0 0 0 00 N 0 NH NH NH N N Rn NH ~ Rn N NH Rn N 0

N o NH oNNH o o o NH o Rn N RnR N N NH NH NH N N o N o N o O~~ 00 N NH NH NH N Rn Rn Rn 0- N N/ NH Ns o o N N- >NH 0 NHH

o o NH Rn R

N S 0 0 N o NH 0 0 N NH Rn NN Rn N 0 o o NH= o o NH RnN NH Rn NH R N S N o NH S NH

Rn O RnRn Rn

31

0 o0 2023248067 10 Oct 2023

o O0 O0 O0 O0 HN HN HN N NH o "IIIIIN N o O HN HN HN NH NN S 0 NH Rn O RA O R = N 0== NIIIi"". 0N->

O O S O O 7f NH Ak o NH NH

HN HN 0 HN Rn 0R0 Rn o N O N o o 0 0 0 0 0 Alk HN HN Alk HN NH Rn NNH Rh NNH O O o R 0= NI i"' N - 0= N - S O O o 0 0 A NH Ak NH NH

Rn 0 Rn 0 HN R HN HN N S N O N O o 0 S 0 0 0 HN HN HN NNH .~NH ,N~ NH Rn Rn Rn O O O N- o0= N- S= N 0== o O O o NH P- O O NH NNH N HN N Rn 0 HN Rn 0 Rnp0 HN N N o N O N o o 0 HN 0 0 HN N 0 0 HN

NNH Rh NNH NH o Rn O o R 0== = 0 N 0 N >=r NN O O O NH ;NH o o O N Rn N 0n Rn 0 HN Rn N 0n HN HN N N O N o N o 0 OS 0 NH0 N 0 N/0 0 N/O HN HN HN O2 Rn Rn o O o 0== R N- 0= N 0== N

SNH N o o NH o o NHHo o O

Rn 0Rn HN: 0 N Rn 0 N

N o N o N o O 00 0 NH 00 OH HN HN HN Rn Rn O Rn o

O o NH pNH o o RnRn HN 0 Rn N 0

o N o

HN HN Rn Rn O o

32

H HN H ZI HHN 0 O N 0 o 0 N N 0 o 0 N 0 o o o 0 0 S S o o MIIIIIIII

N N N N""

Rn Rn H HN Rn Rn H ZI Rn Rn H HN

0 O N 0 0 N 0 S S N 0 O O o o 0 o 0 0 o o All 2023248067

N N NN"""N N 5 N N Alk Alk Alk Alk

Rn H HN Rn OH Rn HN Hn Rn Rn Rn 0 N S S O 0O N o 0 00 o o o N O o ON o 0 0 :

N N N

Rn Rn HN H Rn Rn 0 N 0H o HN HHN O o Rn o o o o 0 0 o o 0 n N N N N N N N N: N N N NNN~

IZ Rn 0 H Rn 0 N 0 Rn HZI Rn H o o Rn Rn HN

o 0 0 o 0 o 0No 0o o o 0 N N. N N rNN N. N N N N N

N N Rn Rn N N Rn Rn Rn Rn HZI HHN o o o j00 o o 0 N 0 o 0 N o 0O : NN o o N D N N N N NN N N. N N N Rn Rn R Rn RRn Rn

33

Oct 2023

H HN H HN

N 0 o 0 o NN 0 o 0 N o 0 0 o 0 2023248067 10 o o

N N NN N N

Rn Rn Rn Rn Rn Rn

H HN

o0 N HO o O o

Rn Rn

N o o o o NH NH 0 Rn Rn N HN 0 o Rn Rn N N HN o

0 o 0 o N 0 0 N o O

N NNH NH NH NH Rn Rn N HN HN 0 o Rn Rn N N H HN 0 o

o 0o NH

NH Rn Rn N N N N HN HN 0 o

termterm

[0063] TheThe

[0063] "independently" "independently" is used is used herein herein to indicate thatthe to indicatethat thevariable, which isis variable, which independently applied, varies independently applied, varies independently fromapplication independently from applicationtoto application. application.

[0064]

[0064] The term"alkyl" The term "alkyl" shall shall meanmean withinwithin its context its context a linear, a linear, branch-chained branch-chained or cyclic fully or cyclic fully

saturated saturatedhydrocarbon hydrocarbon radical radicalororalkyl group, alkyl preferably group, a CI-C1o, preferably a C-C, more more preferably preferablya aC1-C6, C-C,

alternatively alternatively aa C-C CI-C3 alkyl alkyl group, group, which which may may be be optionally optionally substituted. substituted. Examples Examples of alkyl of alkyl

groups aremethyl, groups are methyl,ethyl, ethyl, n-butyl, n-butyl, sec-butyl, sec-butyl, n-hexyl, n-hexyl, n-heptyl, n-heptyl, n-octyl, n-octyl, n-nonyl, n-nonyl, n-decyl, n-decyl,

34

isopropyl, 2-methylpropyl, cyclopropyl, isopropyl, 2-methylpropyl, cyclopropyl,cyclopropylmethyl, cyclopropylmethyl, cyclobutyl, cyclobutyl, cyclopentyl, cyclopentyl, cyclopen cyclopen-

tylethyl, cyclohexylethyl tylethyl, cyclohexylethyl and and cyclohexyl, among amongothers. others.InIncertain certain embodiments, embodiments,thethe alkylgroup alkyl group is is end-capped witha ahalogen end-capped with halogen group group (At,(At, Br, Br, Cl, Cl, F, I). F, or or I). In certain In certain preferred preferred embodiments, embodiments,

compounds according compounds according totothe thepresent present disclosure disclosure which which may maybebeused used to to covalentlybind covalently bindtoto dehalogenase enzymes.These dehalogenase enzymes. These compounds compounds generally generally contain contain a sidea chain side chain (often(often linked linked through through a a polyethylene glycol polyethylene glycol group) group)which which terminates terminates in in an an alkyl alkyl group group which which has has a halogen a halogen substituent substituent

(often (often chlorine chlorine or bromine) on its its distal distal end end which results in in covalent covalent binding of the the compound compound 2023248067

bromine) on which results binding of

containingsuch containing such a moiety a moiety to protein. to the the protein.

[0065] The term

[0065] The term "Alkoxy" "Alkoxy" refers refers to an to an alkyl alkyl groupgroup singularly singularly bonded bonded to oxygen. to oxygen.

[0066] The term

[0066] The term "Alkenyl" "Alkenyl" refers refers to linear, to linear, branch-chained branch-chained or cyclic or cyclic C2-C10 (preferably C2-C (preferably C2- C2 C hydrocarbon C)6 )hydrocarbon containingatatleast radicalscontaining radicals least one C=C bond. bond.

[0067] The term

[0067] The term "Alkynyl" "Alkynyl" refers refers to linear, to linear, branch-chained branch-chained or cyclic or cyclic C2-C10 (preferably C-C (preferably C- C2 C) hydrocarbon C 6 )hydrocarbon containingatatleast radicalscontaining radicals least one bond. C--Cbond.

[0068] The term

[0068] The term "alkylene" "alkylene" when refers when used, used, refers to a -(CH2)n- to a -(CH)n- group (ngroup is an (n is an integer integer generally generally

from 0-6), which from 0-6), which may maybebeoptionally optionallysubstituted. substituted. When When substituted,thethealkylene substituted, alkylenegroup group preferably preferably

is is substituted substituted on on one or more one or moreofofthe themethylene methylene groups groups withwith a C-Ca alkyl C 1-C group alkyl(including group (including a a cyclopropyl group cyclopropyl groupororaa t-butyl t-butyl group), group), but but may also be may also be substituted substituted with with one one or or more morehalo halogroups, groups, preferably from preferably from1 1toto3 3halo halogroups groupsor or oneone or or twotwo hydroxyl hydroxyl groups, groups, 0-(C-C O-(C-C alkyl) or alkyl) 6groups groups or amino acidsidechains amino acid sidechainsasas otherwise otherwisedisclosed disclosedherein. herein. InIncertain certain embodiments, embodiments,an an alkylene alkylene group group

maybebesubstituted may substituted with withaaurethane urethaneororalkoxy alkoxygroup group(or(orother othergroup) group)which which is is furthersubstituted further substituted withaapolyethylene with polyethylene glycol glycol chainchain (of1from (of from to 10,1preferably to 10, preferably 1 to 16,tooften 1 to 6, often 1 to 4glycol 4 ethylene ethylene glycol units) to units) to which whichis issubstituted substituted (preferably, (preferably, but but not exclusively not exclusively on theon the end distal distal end of the of the polyethylene glycol polyethylene glycol chain) chain)ananalkyl alkylchain chainsubstituted substitutedwith witha asingle singlehalogen halogengroup, group,preferably preferably a a chlorine group. group. In In stillother still otherembodiments, embodiments,the the alkylene alkylene (often, (often, a methylene) a methylene) group, group, may may be be substituted with ananamino substituted with amino acid acid sidechain sidechain group group such such as as a sidechain a sidechain group ofgroup of aornatural or a natural

unnatural amino unnatural aminoacid, acid, forfor example, example, alanine, alanine, p-alanine, arginine, ß-alanine, arginine, asparagine, asparagine, aspartic aspartic acid, acid, cysteine, cystine,glutamic cysteine, cystine, glutamic acid, acid, glutamine, glutamine, glycine, glycine, phenylalanine, phenylalanine, histidine,histidine, isoleucine,isoleucine, lysine, lysine, leucine, methionine, leucine, methionine, proline, proline, serine, serine, threonine, threonine, valine, valine, tryptophan tryptophan or tyrosine. or tyrosine.

[0069] The term

[0069] The term "unsubstituted" "unsubstituted" shall shall mean substituted mean substituted only hydrogen only with with hydrogen atoms. Aatoms. range A range of carbon of atomswhich carbon atoms whichincludes includesC means Co means that that carbon carbon is absent is absent andreplaced and is is replaced with with H. Thus, H. Thus, a a

35 range of of carbon atomswhich whichisisCo-C6 includes carbons atomsatoms of 1,of 2,1,3, 2, 4, 5 and 3, 54,and 6 and for for C, Co, H 2023248067 10 Oct 2023 range carbon atoms C-C includes carbons 6 and H stands in place stands in placeofofcarbon. carbon.

[0070] The term

[0070] The "substituted" term "substituted" or "optionally or "optionally substituted" substituted" shallindependently shall mean mean independently (i.e., (i.e., where more where morethan thansubstituent substituentoccurs, occurs,each eachsubstituent substituentisisindependent independentof of another another substituent)oneone substituent)

or more substituents (independently up to five substitutents, preferably up to three substituents, or more substituents (independently up to five substitutents, preferably up to three substituents,

often 1 or often 1 or 22 substituents substituentson on aamoiety moiety in in aa compound accordingtotothe compound according thepresent presentdisclosure disclosure and andmay may include substituents which include substituents whichthemselves themselves maymay be further be further substituted) substituted) at a carbon at a carbon (or nitrogen) (or nitrogen)

position anywhere position anywhereonona molecule a molecule within within context, context, and and includes includes as substituents as substituents hydroxyl, hydroxyl, thiol,thiol,

carboxyl, cyano carboxyl, cyano(C=N), (C-N),nitro nitro(NO), (NOhalogen 2 ), halogen (preferably, (preferably, 1, 2 1,or23 or 3 halogens, halogens, especially especially on anon an alkyl, especially alkyl, especially aa methyl group such methyl group suchasasa atrifluoromethyl), trifluoromethyl), an an alkyl alkyl group group(preferably, (preferably, C-C, C-Cio, morepreferably, more preferably, C-C), 6), aryl CI-C aryl (especially (especially phenyl phenyl and and substituted substituted phenyl phenyl for example for example benzylbenzyl or or benzoyl), alkoxy benzoyl), alkoxy group group(preferably, (preferably,CI-C 6 alkyl C-C alkyl or or aryl,including aryl, includingphenyl phenyl andand substitutedphenyl), substituted phenyl), thioether (C thioether 1-Calkyl (C-C 6 alkyl ororaryl), aryl), acyl acyl (preferably, (preferably,CC1-C 1 -C6acyl), acyl), ester ester or or thioester thioester (preferably,C -C (preferably, 1C1-C

alkyl or aryl) including alkylene ester (such that attachment is on the alkylene group, rather than alkyl or aryl) including alkylene ester (such that attachment is on the alkylene group, rather than

at the ester function which is preferably substituted with aC1 -C6 alkyl or aryl group), preferably, at the ester function which is preferably substituted with a C-C alkyl or aryl group), preferably,

6 alkyl Ci-Calkyl C-C or or aryl, aryl, halogen halogen (preferably, or Cl), F orF C1), (preferably, amine amine (including (including a five- a five- or six-membered or six-membered

cyclic alkylene cyclic alkylene amine, amine, further further including includinga aC -C 6 alkyl C-C1 alkyl amineamine or adialkyl or a C-C C1 -C6 amine dialkylwhich amine which alkyl groups alkyl maybebesubstituted groups may substitutedwith withone oneorortwo twohydroxyl hydroxyl groups) groups) or or an an optionally optionally substituted substituted - -

N(C-C group group alkyl)alkyl) 6 alkyl)C(O)(0-C1-C6 N(Co-Calkyl)C(O)(O-C-C may be may (which (which be optionally optionally substituted substituted with a with a polyethylene glycol polyethylene glycol chain chaintoto which whichisisfurther further bound boundananalkyl alkylgroup groupcontaining containinga single a singlehalogen, halogen, preferably chlorine preferably chlorine substituent), substituent), hydrazine, amido, which hydrazine, amido, whichisispreferably preferablysubstituted substitutedwith withoneone or or two C-C two Ci-C 6 alkyl alkyl groups groups (including (including a carboxamide a carboxamide whichwhich is optionally is optionally substituted withwith substituted one one or or two two

6 alkyl groups), alkanol (preferably, Ci-C 6 alkyl or aryl), or alkanoic acid (preferably,C-C6 CI-Calkyl C-C groups), alkanol (preferably, C-C alkyl or aryl), or alkanoic acid (preferably, C1-C

alkyl or alkyl or aryl). aryl). Substituents Substituentsaccording accordingto tothethe present present disclosure disclosure may may include, include, for example for example -

SiRi subR2subR3subgroups SiRsubRsubR3sub where groups where each each of Risuband of Rub R2su and Rub is 1 is as otherwise as otherwise described described hereinand herein and Rub R3sub

is is H or aaCi-C H or alkylgroup, C1-C6 alkyl group,preferably preferablyRub, Risub, Rub,R2sub, thisincontext R3su Rub in this context is aC1-C3 is a C1-C alkyl group alkyl group

(including an isopropyl (including an isopropyl orort-butyl t-butyl group). group). Each Each of of thethe above-described above-described groups groups may may be be linked linked

directly to the substituted moiety or alternatively, the substituent may be linked to the substituted directly to the substituted moiety or alternatively, the substituent may be linked to the substituted

moiety (preferably in the case of an aryl or heteraryl moiety) through an optionally substituted moiety (preferably in the case of an aryl or heteraryl moiety) through an optionally substituted -

(CH ororalternatively 2 )m- (CH)- alternatively an an optionally optionallysubstituted -(OCH substituted 2 )m-, -(OCH -(OCH)m-, 2 CH2)m- or -(OCHCH)m- -(CH 2 CH2 0)m or -(CHCHO)m- group, whichmaymay group, which be substituted be substituted withwith any orone any one or ofmore more of the above-described the above-described substituents. substituents.

36

Alkylene groups-(CH)m- or -(CH2)n- groups or other chains such as ethylene glycolglycol chains, as 2023248067 10 Oct 2023

Alkylene groups -(CH2)m- or -(CH)n- groups or other chains such as ethylene chains, as

identified identified above, above, may besubstituted may be substituted anywhere anywhereonon thethe chain.Preferred chain. Preferred substitutentsonon substitutents alkylene alkylene

groups include halogen groups include halogen or or C 1-C (preferably C-C 6 (preferably C-C) 3 ) alkyl C1-Calkyl groups,which groups, which maymay be optionally be optionally

substituted with substituted one or with one or two twohydroxyl hydroxylgroups, groups,oneone or or twotwo ether ether groups groups (0-C1-C (O-C-C 6 groups), groups), up to up to three halo groups three groups (preferably (preferably F), F), or or aa sidechain of of an an amino aminoacid acidasasotherwise otherwisedescribed describedherein herein and optionally substituted and optionally substituted amide amide(preferably (preferablycarboxamide carboxamide substituted substituted as described as described above)above) or or urethane groups urethane groups(often (often with withone oneorortwo twoC-C Co-C6 alkyl alkyl substitutents, substitutents, which which group(s) group(s) may may be further be further

substituted). In substituted). In certain certain embodiments, embodiments,thethealkylene alkylene group group (often (often a single a single methylene methylene group) group) is is substituted substituted with one one orortwo twooptionally optionallysubstituted substitutedC-CC 1alkyl -C6 alkyl groups, groups, preferably preferably C1 -C 4 alkyl C-C alkyl

group, mostoften group, most oftenmethyl methyl or or 0-methyl O-methyl groups groups or a sidechain or a sidechain of an acid of an amino amino acid as otherwise as otherwise

described herein. In described herein. In the the present present disclosure, disclosure, aamoiety moiety in in aa molecule molecule may beoptionally may be optionally substituted substituted with up with uptoto five fivesubstituents, substituents, preferably preferably upuptotothree threesubstituents. substituents.Most Most often, often, in present in the the present disclosure, moieties disclosure, moieties which which are substituted are substituted are substituted are substituted with with one onesubstituents. or two or two substituents.

[0071] The term

[0071] The term "substituted" "substituted" (each (each substituent substituent beingbeing independent independent of any of any substituent) other other substituent) shall shall also also mean within its mean within its context context of of use use C-C C 1-C alkyl, alkyl, 6 C-C C 1-C6 alkoxy,alkoxy, halogen, halogen, amido, amido,

carboxamido, sulfone, including carboxamido, sulfone, including sulfonamide, sulfonamide, keto, keto, carboxy, carboxy, C-CC 1ester -C6 ester (oxyester (oxyester or or carbonylester), carbonylester),C-C keto, urethane C-C keto, urethane -0-C(O)-NRsubRsub -- C(O)-NRisubR2sbor or -N(Risub)-C(O)-O-Risub,nitro, -N(R1su)-C(O)-O-R1sub, nitro, cyano andamine cyano and amine(especially (especiallyincluding includinga aC-C C-C alkylene-NRisubR2sub, alkylene-NR1subR2sub, a mono- a mono- or di-orC-C di-alkyl C1 -C6 alkyl

substituted substituted amines whichmay amines which maybebe optionallysubstituted optionally substitutedwith withone oneorortwo two hydroxyl hydroxyl groups). groups). EachEach

of these these groups contain unless groups contain unless otherwise indicated, indicated, within within context, context, between between 11 and and 66 carbon carbon atoms. atoms. In certain In certain embodiments, preferredsubstituents embodiments, preferred substituentswill will include includefor forexample, example,-NH-, -NH-, -NHC(O)-, -NHC(O)-, -0-, -0-, -(CH 2 )m- =0, -(CH)m- =0, (here, m and (here,m and n are n are in context, in context, 1, 1, 3, 3,4,4,5 5oror6), 2, 2, 6), -S-, -S-, -S(O)-, -S(O)-, SO- S02-oror-NH-C(O)- -NH-C(O) NH-, -(CH2)nOH, NH-, -(CH)nOH, -(CH2)nSH, -(CH2)nCOOH, -(CH)SH, -(CH)COOH, Ci-C6 C1-C alkyl, -(CH)O-(C-C alkyl, -(CH2)nO-(C1-C6 alkyl), -(CH)C(O)- alkyl), -(CH2)nC(O)

(Ci-C alkyl), -(CH (C-C 6alkyl), 2 )nOC(O)-(C1-C 6alkyl), -(CH)OC(O)-(C-C alkyl), -(CH 2 )nC(0)0-(C1-C 6 alkyl), -(CH),C(O)O-(C-C alkyl), -(CH 2)nNHC(O)-Risub, -(CH)NHC(O)-Rb, -(CH 2)nC(O)-NRisubR2sub, -(OCH 2)nOH, -(CHO)COOH, -(OCH)OH, -(CH 20)nCOOH,C-C C 1-C 6 alkyl, -(OCH alkyl, 2 )nO-(C1-C 6 alkyl), -(OCH)O-(C-C -(CH 2 0)nC(O)-(C1-C 6 alkyl), -(OCH 2)nNHC(O)-Risub, -(CH 2 0)nC(O)-NRisubR2sub, -S(0)2-Rs, -(CHO)C(O)-(C-C alkyl), -(OCH)NHC(O)-R1sb, -S(O)-Rs, - S(O)-Rs(Rs S(O)-Rs (RsisisC1-C 6 alkyl C-Calkyl or or a -(CH2)m-NRisubR2sub a -(CH2)m-NRsubRsub group), NO, CNNO group), , CN or(F, or 2halogen halogen (F,I,Cl, Cl, Br, Br, I, preferably FF or preferably or C1), Cl), depending dependingononthe thecontext contextofofthe theuse useofofthe thesubstituent. substituent. Rub Risub and and Rub R2sub are are each, within within context, context, H or aa C H or 1 -Calkyl C-C 6 alkyl group group (which (which may may be optionally be optionally substituted substituted withwith one or one or

two hydroxyl two hydroxylgroups groupsororupuptotothree threehalogen halogengroups, groups,preferably preferablyfluorine). fluorine). The The term term "substituted" "substituted"

shall shall also also mean, within the mean, within the chemical chemicalcontext contextofofthe thecompound compound defined defined and and substituent substituent used, used, an an

37 optionally substituted aryl or heteroaryl groupgroup or an optionally substituted heterocyclic group as 2023248067 10 Oct 2023 optionally substituted aryl or heteroaryl or an optionally substituted heterocyclic group as otherwise describedherein. otherwise described herein.Alkylene Alkylene groups groups may bealso may also be substituted substituted as otherwise as otherwise disclosed disclosed herein, preferably herein, preferably with with optionally optionally substituted substituted C-C C 1-Calkyl 6 alkyl groups groups (methyl, (methyl, ethyl ethyl or or hydroxymethylor or hydroxymethyl hydroxyethyl hydroxyethyl is preferred, is preferred, thus thus providing providing a chiral a chiral center), center), a sidechain a sidechain of of an an amino acidgroup amino acid groupasasotherwise otherwisedescribed describedherein, herein,anan amido amido group group as described as described hereinabove, hereinabove, or a or a urethane group urethane groupO-C(O)-NRisubR2sub O-C(O)-NR group group where where RisubRub Rub and andare as are R2sub as otherwise otherwise described described herein, although herein, numerousother although numerous other groups groups may may also also be used be used as substituents. as substituents. Various Various optionally optionally substituted substituted moieties maybebesubstituted moieties may substitutedwith with3 3orormore more substituents,preferably substituents, preferablynonomore more than than 3 3 substituents and substituents preferably with and preferably with1 1oror2 2substituents. substituents. It Itisisnoted notedthat thatinininstances instanceswhere, where, in in a a compound compound at aatparticular a particular position position of theofmolecule the molecule substitution substitution is required is required (principally, (principally, because of because of valency), butnono valency), but substitution substitution is indicated, is indicated, then then that that substituent substituent is construed is construed or understood or understood to be H, to be H, unlessthe unless thecontext contextof of thethe substitution substitution suggests suggests otherwise. otherwise.

term "aryl" The "aryl"

[0072] The term

[0072] or "aromatic", or "aromatic", in context, refers torefers in context, to a substituted a substituted (as (as otherwise otherwise described herein) described herein) ororunsubstituted unsubstitutedmonovalent monovalent aromatic aromatic radical radical havinghaving a singlea ring single ring (e.g., (e.g., benzene, phenyl, benzene, phenyl, benzyl) benzyl)ororcondensed condensed rings rings (e.g., (e.g., naphthyl, naphthyl, anthracenyl, anthracenyl, phenanthrenyl, phenanthrenyl, etc.) etc.)

and can and can bebebound boundto to thethe compound compound according according to thetopresent the present disclosure disclosure at anyatavailable any available stablestable

position on position on the the ring(s) ring(s) or or asas otherwise otherwiseindicated indicatedin in thethe chemical chemical structure structure presented. presented. OtherOther

examples ofofaryl examples aryl groups, groups, inincontext, context, may may include include heterocyclicaromatic heterocyclic aromatic ring ring systems, systems,

"heteroaryl" groups "heteroaryl" groups having havingone oneorormore morenitrogen, nitrogen,oxygen, oxygen,ororsulfur sulfuratoms atomsininthe thering ring (moncyclic) (moncyclic) such as imidazole, such as imidazole,furyl, furyl,pyrrole, pyrrole,furanyl, furanyl,thiene, thiene, thiazole, thiazole, pyridine, pyridine, pyrimidine, pyrimidine, pyrazine, pyrazine,

triazole, oxazole triazole, or fused oxazole or fusedring ringsystems systems such such as indole, as indole, quinoline, quinoline, indolizine, indolizine, azaindolizine, azaindolizine,

benzofurazan, etc., benzofurazan, etc., among among others, others, which which may may be be optionally optionally substituted substituted as described as described above. above. Among Among thethe heteroaryl heteroaryl groups groups which which may may be be mentioned mentioned include include nitrogen-containing nitrogen-containing heteroaryl heteroaryl

groups suchas as groups such pyrrole, pyrrole, pyridine, pyridine, pyridone, pyridone, pyridazine, pyridazine, pyrimidine, pyrimidine, pyrazine, pyrazine, pyrazole, pyrazole,

imidazole, triazole, triazole, triazine, triazine, tetrazole, tetrazole, indole, indole, isoindole, isoindole, indolizine, indolizine, azaindolizine, azaindolizine,purine, purine, indazole, quinoline, dihydroquinoline, indazole, quinoline, dihydroquinoline,tetrahydroquinoline, tetrahydroquinoline,isoquinoline, isoquinoline,dihydroisoquinoline, dihydroisoquinoline, tetrahydroisoquinoline, tetrahydroisoquinoline, quinolizine, phthalazine, quinolizine, phthalazine, naphthyridine, naphthyridine, quinoxaline, quinoxaline, quinazoline, quinazoline,

cinnoline, pteridine, cinnoline, pteridine, imidazopyridine, imidazopyridine, imidazotriazine, imidazotriazine, pyrazinopyridazine, pyrazinopyridazine, acridine, acridine, phenanthridine, carbazole, phenanthridine, carbazole,carbazoline, carbazoline,pyrimidine, pyrimidine, phenanthroline, phenanthroline, phenacene, phenacene, oxadiazole, oxadiazole,

benzimidazole, pyrrolopyridine, benzimidazole, pyrrolopyridine,pyrrolopyrimidine pyrrolopyrimidine and pyridopyrimidine; and pyridopyrimidine; sulfur-containing sulfur-containing

aromatic heterocycles aromatic heterocycles such as thiophene such as thiophene and and benzothiophene; benzothiophene; oxygen-containing oxygen-containing aromatic aromatic

38 heterocycles such such as as furan, furan, pyran, cyclopentapyran, benzofuran benzofuranand andisobenzofuran; isobenzofuran; andand aromatic 2023248067 10 Oct 2023 heterocycles pyran, cyclopentapyran, aromatic heterocycles comprising heterocycles comprising2 or 2 or more more hetero hetero atoms atoms selected selected fromnitrogen, from among among nitrogen, sulfur andsulfur and oxygen, suchasasthiazole, oxygen, such thiazole, thiadizole, thiadizole, isothiazole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole, benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, isoxazole, phenothiazine, isoxazole, furazan, furazan, phenoxazine, phenoxazine,pyrazoloxazole, pyrazoloxazole, imidazothiazole, imidazothiazole, thienofuran, thienofuran, furopyrrole, furopyrrole, pyridoxazine, furopyridine, furopyridine, furopyrimidine, furopyrimidine,thienopyrimidine thienopyrimidineandand oxazole, oxazole, among among others, all of others, all of which may which may be optionally be optionally substituted. substituted.

The term

[0073] The term

[0073] aryl" aryl" "substituted "substituted refers refers an aromatic to antoaromatic carbocyclic group group carbocyclic comprised comprised of at of at least least one aromatic ring one aromatic ring ororofofmultiple multiplecondensed condensed rings rings at least at least oneone of which of which beingbeing aromatic, aromatic,

wherein the wherein the ring(s) ring(s) are are substituted substitutedwith withone one or ormore more substituents. substituents. For For example, an aryl example, an aryl group can group can

comprise aa substituent(s) comprise substituent(s)selected from:from: selected -(CH -(CH)nOH, 2)nOH, -(CH2)n-O-(C1-C 6)alkyl, -(CH -(CH)n-O-(C-C)alkyl, 2 )n-O-(CH 2 )n -(CH)-O-(CH)n-

(C-C)alkyl, -(CH)-C(O)(C-C) (C1-C6)alkyl, alkyl,-(CH)-C(O)O(C-C)alkyl, -(CH2)n-C(O)(Co-C6)alkyl, -(CH2)n-OC(O)(Co -(CH)n-OC(O)(C- -(CH2)n-C(O)O(Co-C6)alkyl,

C6)alkyl, C)alkyl, amine, or di-(C1-C mono- or amine, mono- di-(C-C 6alkyl) wherein the amine wherein alkyl) amine alkyl group the alkyl group on amine isis the amine on the optionally substituted with optionally substituted with 11 or or 22 hydroxyl groups or hydroxyl groups or up up toto three three halo halo (preferably (preferably F, F, C1) Cl) groups, groups,

OH, COOH, OH, COOH, Ci-C C1-C 6 alkyl, alkyl, preferably CH, preferably CH 3CF, , CFOMe, 3 , OMe, OCF,OCF NO,3 , or NOCN 2 , or CN (each group group of (each of which which

maybebesubstituted may substituted inin ortho-, ortho-, meta- meta- and/or and/orpara- para- positions positions ofofthe the phenyl phenylring, ring, preferably preferably para-), para-), an optionally optionally substituted substituted phenyl phenyl group group(the (thephenyl phenylgroup group itselfisispreferably itself preferablysubstituted substituted with witha a linker linker group group attached to to aa PTM group,including PTM group, includinga aULM ULM group), group), and/or and/or at leastoneone at least of of F,F, Cl,OH, Cl, OH, COOH, COOH, CH,CH 3 , OMe, CF, CF3 , OCF, OMe,NO, OCF , NO or 3CN 2 , or(in group CNortho-, group (in ortho-, meta- meta- and/or and/or para- para-of positions positions the of the phenyl ring, phenyl ring, preferably preferablypara-), para-),a anaphthyl naphthyl group, group, whichwhich may bemay be optionally optionally substituted, substituted, an an optionally substituted heteroaryl, optionally substituted heteroaryl, preferably preferably ananoptionally optionallysubstituted substituted isoxazole isoxazole including including a a methylsubstituted isoxazole, ananoptionally methylsubstituted isoxazole, optionallysubstituted substitutedoxazole oxazole including including a methylsubstituted a methylsubstituted

oxazole, oxazole, anan optionally optionally substituted substituted thiazole thiazole including including a methyla substituted methyl substituted thiazole, thiazole, an an optionally optionally

substituted isothiazole including substituted isothiazole including a amethyl methyl substituted substituted isothiazole, isothiazole, an optionally an optionally substituted substituted

pyrrole including pyrrole including aa methylsubstituted methylsubstitutedpyrrole, pyrrole,ananoptionally optionallysubstituted substitutedimidazole imidazole including including a a methylimidazole, an methylimidazole, an optionally optionally substituted substituted benzimidazole benzimidazole or methoxybenzylimidazole, an or methoxybenzylimidazole, an optionally optionally substituted substituted oximidazole or methyloximidazole, oximidazole or methyloximidazole, ananoptionally optionallysubstituted substituted diazole diazole group, group, including a methyldiazole including a methyldiazole group, group, ananoptionally optionally substituted substituted triazole triazole group, group, including including aa methylsubstituted triazole group, methylsubstituted triazole group, ananoptionally optionallysubstituted substitutedpyridine pyridine group, group, including including a halo a halo-

(preferably, (preferably, F) F) or or methylsubstitutedpyridine group ororananoxapyridine methylsubstitutedpyridine group oxapyridinegroup group (where (where the the pyridine pyridine

group is linked to group is to the the phenyl phenyl group by an group by anoxygen), oxygen),ananoptionally optionallysubstituted substituted furan, furan, an an optionally optionally substituted substituted benzofuran, anoptionally benzofuran, an optionallysubstituted substituteddihydrobenzofuran, dihydrobenzofuran, an optionally an optionally substituted substituted

39

indole, indolizineor orazaindolizine indole, indolizine azaindolizine (2,or3,4-azaindolizine), (2, 3, or 4-azaindolizine), an optionally an optionally substituted substituted quinoline,quinoline,

and combinationsthereof. and combinations thereof.

[0074] "Carboxyl"

[0074] "Carboxyl" denotes thethe denotes group group --C(O)OR, --C(O)OR, where where is hydrogen, R isRhydrogen, alkyl, alkyl, substituted substituted

alkyl, aryl, substituted alkyl, aryl, substituted aryl, aryl, heteroaryl heteroaryl ororsubstituted substitutedheteroaryl heteroaryl , whereas , whereas these these genericgeneric

substituents substituents have meaningswhich have meanings which are are identical identical withwith definitions definitions of corresponding of the the corresponding groupsgroups

defined herein. defined herein.

2023248067

[0075] The term

[0075] The term "heteroaryl"or "heteroaryl"or "hetaryl" "hetaryl" can but can mean meanis but is in in no waynolimited way limited to an optionally to an optionally

substituted substituted quinoline quinoline (which maybebeattached (which may attachedtotothe the pharmacophore pharmacophoreor or substitutedon on substituted anyany carbon carbon

atom withinthe atom within thequinoline quinolinering), ring), ananoptionally optionallysubstituted substitutedindole indole(including (includingdihydroindole), dihydroindole),an an optionally substitutedindolizine, optionally substituted indolizine, an optionally an optionally substituted substituted azaindolizine azaindolizine (2, 3 or (2, 3 or 4-azaindolizine) 4-azaindolizine)

an optionally substituted an optionally substituted benzimidazole, benzimidazole, benzodiazole, benzodiazole,benzoxofuran, benzoxofuran, an optionally an optionally substituted substituted

imidazole, anoptionally imidazole, an optionallysubstituted substitutedisoxazole, isoxazole,an an optionally optionally substituted substituted oxazole oxazole (preferably (preferably

methyl substituted), methyl substituted), ananoptionally optionallysubstituted substituteddiazole, diazole, an optionally an optionally substituted substituted triazole, triazole, a a tetrazole, an optionally substituted benzofuran, an optionally substituted thiophene, an optionally tetrazole, an optionally substituted benzofuran, an optionally substituted thiophene, an optionally

substituted thiazole (preferably substituted thiazole (preferably methyl methyl and/or and/or thiol thiol substituted), substituted), an optionally an optionally substituted substituted

isothiazole, isothiazole, an optionally substituted an optionally substituted triazole triazole (preferably (preferably a a1,2,3-triazole 1,2,3-triazole substituted substitutedwith witha a methyl group, aa triisopropylsilyl methyl group, triisopropylsilyl group, group, an optionallysubstituted an optionally substituted-(CH 2 )m-0-C 1 -C -(CH)n-O-C-C 6 alkyl alkyl group group or or an optionally substituted an optionally substituted -(CH 2 )m-C(O)-O-CI-C -(CH)C(O)-O-C-C 6 alkyl group), alkyl group), an optionally an optionally substituted substituted pyridinepyridine (2-, (2-, 3, 3, or or 4-pyridine) 4-pyridine) orora agroup group according according to chemical to the the chemical structure: structure:

SSC sc O H RHET O O - -== K , - RHET R HET 2 N N N N \URE URE URE R R URE 0 0 o o HET RHET RHET N R N-3 RHET HET R HET N R N N ONN 0 O

HET K-JN RHET R YC wherein wherein

40

Se is Sc is CHRSS, NRu", CHRSS, NR or O; URE, or 0; 2023248067 10 Oct 2023

RT is is RHET H, H, CN,CN, NO,N02, halo halo (preferably (preferably Cl orClF), or optionally F), optionally substituted substituted C-C C-C6 alkyl (preferably alkyl (preferably

substituted with substituted with one oneorortwo two hydroxyl hydroxyl groups groups or up or to up to halo three threegroups halo (e.g. groupsCF), (e.g. CF3 ),

optionally substituted substituted O(Ci-C 6 alkyl) O(C1-C alkyl) (preferably (preferably substitutedwith substituted with oneone or two or two hydroxyl hydroxyl

groups or up groups or up to to three three halo halo groups) or an groups) or optionally substituted an optionally substituted acetylenic acetylenic group group -C=C-Ra -C=C-Ra

where RRaisisH Horora aC-C where CI-C 6 alkyl alkyl group group (preferably (preferably C-C C-C3 alkyl); alkyl);

Rss is H, RSS is H, CN, N02, CN, NO, halo halo (preferably (preferably F or F or Cl), C1), optionally optionally substitutedC-CC-C substituted alkylalkyl 6 (preferably (preferably

substituted with substituted with one oneorortwo two hydroxyl hydroxyl groups groups or uportoup to three three halo groups), halo groups), optionally optionally

substituted 0-(CI-C substituted 6 alkyl) O-(C-C alkyl) (preferablysubstituted (preferably substitutedwith withone oneorortwo twohydroxyl hydroxyl groups groups or or up up to three to three halo halo groups) groups) or or an optionally substituted an optionally substituted -C()(C-C6 alkyl) (preferably -C(O)(C1-C alkyl) (preferably substituted with substituted with one one or two two hydroxyl groupsororup hydroxyl groups up to to three three halo groups); RURE RUl is H, aa C is H, C-C 6 alkyl (preferably H or C1 -C 3 alkyl) or a -C(O)(CI-C 6 alkyl), each of 1 -Calkyl (preferably H or C-C alkyl) or a -C(O)(C-C alkyl), each of

whichgroups which groupsisisoptionally optionallysubstituted substitutedwith withone oneor or two two hydroxyl hydroxyl groups groups or uportoup to three three

halogen, preferably halogen, preferably fluorine fluorine groups, groups, or or an an optionally optionally substituted substituted heterocycle, heterocycle,for forexample example

piperidine, morpholine, piperidine, morpholine,pyrrolidine, pyrrolidine,tetrahydrofuran, tetrahydrofuran,tetrahydrothiophene, tetrahydrothiophene, piperidine, piperidine,

piperazine,each piperazine, eachof of which which is optionally is optionally substituted, substituted, and and YCisis NNororC-R Y C-Rc, where , where RYc R is H, isOH, H, CN, OH,NO, CN,halo N02, halo (preferably (preferably Cl oroptionally Cl or F), F), optionally substituted Ci-C substituted 6 alkyl C-C alkyl (preferablysubstituted (preferably substitutedwith withone one or or two two hydroxyl hydroxyl groups groups or to or up up to three halo three halo groups (e.g. CF groups (e.g. 3 ), optionally CF), optionally substituted substituted O(C-C alkyl)(preferably O(C1-C 6alkyl) (preferablysubstituted substituted with one with one or or two two hydroxyl hydroxylgroups groups or or up up to to threehalo three halogroups) groups) or or an an optionallysubstituted optionally substituted acetylenic group acetylenic -C=C-Rawhere group -C=C-Ra where RaHisorH aorC-C R is a CI-C 6 alkyl alkyl groupgroup (preferably (preferably C-C3 C-C alkyl). alkyl).

[0076] TheThe

[0076] termterm "Heterocycle" "Heterocycle" refers refers a cyclic to atocyclic group group which which contains contains at least at least one one heteroatom, e.g., heteroatom, e.g., N, N, O 0or or S, S, and and may may be aromatic be aromatic (heteroaryl) (heteroaryl) or non-aromatic. or non-aromatic. Thus, theThus, the heteroaryl moieties heteroaryl moieties are are subsumed subsumedunder under thethe definitionofofheterocycle, definition heterocycle,depending depending on on the the context context

of of its itsuse. use.Exemplary Exemplary heteroaryl heteroaryl groups groups are described described hereinabove. hereinabove.

[0077] Exemplary

[0077] Exemplary heterocyclics heterocyclics include: include: azetidinyl, azetidinyl, benzimidazolyl, benzimidazolyl, 1,4- benzodioxanyl, 1,4- benzodioxanyl,

1,3-benzodioxolyl, benzoxazolyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl, benzothiazolyl,benzothienyl, benzothienyl, dihydroimidazolyl, dihydroimidazolyl,

dihydropyranyl, dihydrofuranyl,dioxanyl, dihydropyranyl, dihydrofuranyl, dioxanyl,dioxolanyl, dioxolanyl,ethyleneurea, ethyleneurea,1,3-dioxolane, 1,3-dioxolane,1,3-dioxane, 1,3-dioxane, 1,4-dioxane, furyl,homopiperidinyl, 1,4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolyl, imidazolinyl, imidazolinyl, imidazolidinyl, imidazolidinyl, indolinyl, indolyl, indolinyl, indolyl,

isoquinolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolidinyl, isoxazolyl, isoxazolyl, morpholinyl, morpholinyl,

naphthyridinyl, oxazolidinyl, oxazolyl, naphthyridinyl, oxazolidinyl, oxazolyl,pyridone, pyridone, 2-pyrrolidone, 2-pyrrolidone, pyridine, pyridine, , N- , N piperazinyl, piperazinyl,

41 methylpiperazinyl, piperidinyl, piperidinyl,phthalimide, phthalimide, 2023248067 10 Oct 2023 methylpiperazinyl, succinimide, succinimide, pyrazinyl, pyrazinyl, pyrazolinyl, pyrazolinyl, pyridyl,pyridyl, pyrimidinyl,pyrrolidinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolinyl, pyrrolyl, pyrrolyl, quinolinyl, quinolinyl, tetrahydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyranyl, tetrahydroquinoline, thiazolidinyl, tetrahydroquinoline, thiazolidinyl, thiazolyl, thiazolyl, thienyl, thienyl, tetrahydrothiophene, tetrahydrothiophene,oxane, oxane, oxetanyl, oxetanyl, oxathiolanyl, thiane oxathiolanyl, thiane among others. among others.

[0078] Heterocyclic

[0078] Heterocyclic groups can becan groups be optionally optionally substituted with awith substituted member selected selected a member from the from the group consisting of group consisting ofalkoxy, alkoxy, substituted substituted alkoxy, alkoxy, cycloalkyl, cycloalkyl, substituted substituted cycloalkyl, cycloalkyl, cycloalkenyl, cycloalkenyl, substituted cycloalkenyl, substituted cycloalkenyl, acyl, acyl,acylamino, acylamino, acyloxy, acyloxy, amino, amino, substituted substituted amino, amino, aminoacyl, aminoacyl,

aminoacyloxy, oxyaminoacyl, azido, aminoacyloxy, oxyaminoacyl, azido, cyano, cyano,halogen, halogen,hydroxyl, hydroxyl,keto, keto,thioketo, thioketo,carboxy, carboxy, carboxyalkyl, thioaryloxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheteroaryloxy, thioheterocyclooxy, thioheterocyclooxy,thiol, thiol,thioalkoxy, thioalkoxy,substituted substituted thioalkoxy, aryl, thioalkoxy, aryl, aryloxy, aryloxy, heteroaryl, heteroaryl,heteroaryloxy, heteroaryloxy,heterocyclic, heterocyclic, heterocyclooxy, heterocyclooxy,

hydroxyamino, alkoxyamino, hydroxyamino, alkoxyamino, nitro, nitro, -SO-alkyl, -SO-alkyl, -SO-substituted -SO-substituted alkyl, alkyl,-SOaryl, -SOaryl,-SO--SO heteroaryl, -S02-alkyl, heteroaryl, -SO2-substituted -SO2-alkyl, -SO2-substituted alkyl, alkyl, -S02-aryl, -SO2-aryl, oxo and OXO (=0), (=0), and -S02-heteroaryl. -SO2-heteroaryl.

Such heterocyclic Such heterocyclicgroups groupscancan havehave a single a single ring ring or multiple or multiple condensed condensed rings. Examples rings. Examples of of nitrogen heterocycles nitrogen heterocyclesandand heteroaryls heteroaryls include, include, but not but are are limited not limited to, pyrrole, to, pyrrole, imidazole, imidazole,

pyrazole, pyridine, pyrazole, pyridine, pyrazine, pyrazine, pyrimidine, pyrimidine,pyridazine, pyridazine, indolizine, indolizine, isoindole, isoindole, indole, indole, indazole, indazole,

purine, quinolizine, purine, quinolizine, isoquinoline, isoquinoline,quinoline, quinoline, phthalazine, phthalazine, naphthylpyridine, naphthylpyridine, quinoxaline, quinoxaline,

quinazoline,cinnoline, quinazoline, cinnoline, pteridine, pteridine, carbazole, carbazole, carboline, carboline, phenanthridine, phenanthridine, acridine, acridine, phenanthroline, phenanthroline, isothiazole, phenazine, isoxazole, isothiazole, phenazine, isoxazole,phenoxazine, phenoxazine, phenothiazine, phenothiazine, imidazolidine, imidazolidine, imidazoline, imidazoline,

piperidine,piperazine, piperidine, piperazine, indoline, indoline, morpholino, morpholino, piperidinyl, piperidinyl, tetrahydrofuranyl, tetrahydrofuranyl, and and the like as the well like as well as N-alkoxy-nitrogencontaining as N-alkoxy-nitrogen containingheterocycles. heterocycles. TheThe termterm "heterocyclic" "heterocyclic" also also includes includes bicyclic bicyclic

groups in which groups in whichany anyofofthe the heterocyclic heterocyclic rings rings is is fused fused to to aa benzene ring or benzene ring or aa cyclohexane ring or cyclohexane ring or anotherheterocyclic another heterocyclic ring ring (for(for example, example, indolyl, indolyl, quinolyl, quinolyl, isoquinolyl, isoquinolyl, tetrahydroquinolyl, tetrahydroquinolyl, and the and the like). like).

[0079] The term

[0079] The term "cycloalkyl" "cycloalkyl" can but can mean is but mean is way in no way limited in nolimited to univalent to univalent derivedderived groups groups

from monocyclicor or from monocyclic polycyclic polycyclic alkyl alkyl groups groups or cycloalkanes, or cycloalkanes, as defied as defnied herein, herein, e.g.,e.g., saturated saturated

monocyclichydrocarbon monocyclic hydrocarbon groups groups having having fromfrom three three to twenty to twenty carbon carbon atomsatoms in ring, in the the ring, including, including,

but not but not limited limited to, to, cyclopropyl, cyclopropyl, cyclobutyl, cyclobutyl, cyclopentyl, cyclopentyl, cyclohexyl, cyclohexyl,cycloheptyl cycloheptyl andand thethe like. like.

The term The term "substituted "substituted cycloalkyl" cycloalkyl"can can mean but is mean but is in in no no way limited to way limited to aa monocyclic monocyclic or or polycyclic alkyl polycyclic alkyl group groupand andbeing beingsubstituted substitutedbyby one one or or more more substituents, substituents, forfor example, example, amino, amino,

halogen, alkyl, halogen, alkyl, substituted substituted alkyl, alkyl, carbyloxy, carbyloxy,carbylmercapto, carbylmercapto, aryl, aryl, nitro, nitro, mercapto mercapto or sulfo, or sulfo,

42 whereas these these generic genericsubstituent substituent groups groupshave havemeanings meanings which are identical withwith definitions of 2023248067 10 Oct 2023 whereas which are identical definitions of the corresponding the corresponding groups groups as defined as defined in this this legend. inlegend.

[0080]

[0080] The term The term"hydrocarbyl" "hydrocarbyl"shall shallmean mean a compound a compound whichwhich contains contains carbon carbon and hydrogen and hydrogen

and whichmay and which may be be fully fully saturated, saturated, partiallyunsaturated partially unsaturated or or aromatic aromatic and and includes includes aryl aryl groups, groups,

alkyl alkyl groups, groups, alkenyl alkenyl groups and alkynyl groups and alkynyl groups. groups.

[0081] The term

[0081] The term "lower "lower alkyl"alkyl" refers refers to methyl, to methyl, ethylethyl or propyl or propyl

[0082] The term

[0082] The term "lower "lower alkoxy" alkoxy" refersrefers to methoxy, to methoxy, ethoxyethoxy or propoxy. or propoxy.

[0083] More

[0083] More specifically, non-limiting examples specifically,non-limiting CLMs of CLMs examples of include thoseshown includethose shown below below as as well as as "hybrid" "hybrid" molecules moleculesororcompounds compounds that that arise arise fromfrom combining combining 1 or featrues 1 or more more featrues of the of the

following compounds: following compounds:

43

0 0 0 O 0 o O O Rn Rn .1 NH NH HN HN

[ii N SN S O 0 Rn Rn-~ M N o

R' R' 0 O R¹ O (V) () (W) (M)

N N o 0 0 O 1 Rn HN N HN N NH Rn- N O Rn / Nt O R¹

R Mx (x) (y) ()

IZ O O 0 0 N H0 N:i H 0 O N O HN O Rn Rn--..- N o CON CNN w N M R R N

(Z) (z) (aa) (ee)

0 O N HN ~jNN 0 O

RT Rn Rn \/

(ab) (qe)

44

H ZI H 0:1 O N O N o O N0 N NH N R³ RN N N 0 O N Rn-I Rn N N 0 O

2023248067

(ac) (ac) (ad) (ad)

0N 0 o N O N-NH NH R³ R N O Rn NCZ 0 N N O NH0 NH 00 O O

(ae) (ae) (af) (af)

0 /1-NN O

Rn HN- 0 HN O NH NH 0 O (ag) (ag)

45

Oct 2023 o o R5 R 0 o 0 o NH N Q R N o W R' 022 2023248067 10 Rn

O R² o NH NH NH N NH NN 0 o N N o

1 Rn QR W R' R' Rn R' N O R¹

R R o 0 O 0 o 0 o NN NH NIC NH N Nt N o0 o HN HN 0 o CN0 N o R'

H HN 0 o N o 0 o o 0 o

N+ NH NH 0Ny N N CH N o Ho HO 0 O Rn W Rn R R¹ HO HO

wherein: wherein:

Wisis independently W independentlyselected fromthe selectedfrom thegroup groupCH, CHCHR, 2,CHR, C=O, C=O, S02,NH, SO, NH, and N-alkyl; and N-alkyl;

R' is R¹ is selected selected from from the the group group absent, absent, H, H, CH, CN,C1-C3 CH, CN, Cl-C3 alkyl; alkyl;

is HH or R²2 is R or aa Cl-C3 alkyl; C1-C3 alkyl;

is selected R³ is R3 selectedfrom fromH, H, alkyl, alkyl, substituted substituted alkyl, alkyl, alkoxy, alkoxy, substituted substituted alkoxy;alkoxy;

R4 R isis methyl methylororethyl; ethyl; R' R isis HHor or halo; halo; R'isis HHoror halo; R halo;

46

R of CLM isisH; the CLM of the H; 2023248067 10 Oct 2023

R

R' is R' is H or an H or an attachment pointfor attachmentpoint foraaPTM, PTM, a PTM', a PTM', a chemical a chemical linker linker (L), a(L), groupgroup a ULM, ULM, a a CLM, CLM, aa CLM', QI andQ2Q2 Q1 and areare each each independently independently C orCN or N substituted substituted with with a group a group independently independently selected selected

from from HHor or C1-C3 Cl-C3alkyl; alkyl;

is is aa single or double single or doublebond; bond; andand

Rn comprises Rn comprisesa afunctional functional group groupororananatom. atom. R1 , R 2, Qi, Q2, Q3, and Rn Q4, and

[0084]

[0084] In In anyany of of thetheembodiments describedherein, described embodiments the W, herein, the W, R¹, R², Q, Q2, Q, Q4, Rn

can independentlybebecovalently can independently covalentlycoupled coupled to to a linkerand/or a linker and/ora linker a linkertotowhich whichis is attachedoneone attached or or

more PTM, more PTM,ULM, ULM,ULM', ULM', CLMCLM or CLM' or CLM' groups. groups.

[0085] the R¹,the 1, R R²,RQ, 2 , Qi, Q2, and Q4,canand Q3, Rn Rn can

[0085] In any In of anythe of embodiments described the embodiments herein,herein, described Q2, Q, Q4,

independently becovalently independently be covalentlycoupled coupledtotoa alinker linker and/or and/or aalinker linker to to which is attached which is attached one one or or more more PTM, ULM, PTM, ULM,ULM', ULM', CLMCLM or CLM' or CLM' groups. groups.

[0086] In In

[0086] any any of the of the embodiments embodiments described described herein, the the herein, Q, Q2, Q2,Q4, Qi, Q, Q3,andQ4,Rn and can Rn can independently becovalently independently be covalentlycoupled coupledtotoa alinker linker and/or and/or aalinker linker to to which is attached which is attached one one or or more more PTM, ULM, PTM, ULM,ULM', ULM', CLMCLM or CLM' or CLM' groups. groups.

[0087] any aspect In aspect

[0087] In any or embodiment or embodiment described herein,herein, described Rn is modified R is modified to be covalently to be covalently joined joined to the to the linker linkergroup group (L), (L),a aPTM, PTM, a ULM, ULM, a asecond secondCLM CLM having having the same the same chemical chemical structure structure as as the the CLM, CLM, a aCLM', CLM', a second a second linker, linker, or or any any multiple multiple or or combination combination thereof. thereof.

Exemplary

[0088] Exemplary

[0088] Linkers Linkers

[0089] In In

[0089] embodiments,thethecompounds certainembodiments, certain compounds as described as described herein herein include more oneor ormore includeone CLMs chemically CLMs chemically linked linked or or coupled coupled to to oneone or or more more PTMs PTMs (e.g., (e.g., PTM PTM and/orand/or PTM'),PTM'), ULMs (e.g., ULMs (e.g.,

ULM,ULM', ULM, ULM', and/or and/or CLM') CLM') via a via a chemical chemical linkerlinker (L).certain (L). In In certain embodiments, embodiments, the linker the linker group group L is L is aa group group comprising oneoror more comprising one morecovalently covalentlyconnected connected structuralunits structural units (e.g., (e.g., -AL...(AL)q -AL (A¹)q- oror -

(AL)q-), whereinA Ais 1is (A¹)-), wherein a group a group coupled coupled to PTM, to PTM, and and Aq is Aq is a group a group coupledcoupled to at one to at least leastofone a of a ULM,a aULM', ULM, ULM', a CLM, a CLM, a CLM', a CLM', or a combination or a combination thereof.thereof. In certain In certain embodiments, embodiments, AL links ALilinksa a

CLM CLM or or CLM' CLM' directly directly to another to another ULM,ULM, PTM, PTM, or or combination combination thereof. thereof. In other In other embodiments, embodiments,

AL links AL 1 links a CLM or CLM' indirectly to another ULM, PTM, or combination thereof through a CLM or CLM' indirectly to another ULM, PTM, or combination thereof through Aq. Aq.

[0090] In In

[0090] anyany or or aspect aspect embodiment embodiment described described herein, thelinker herein,the group LL isis aa bond linker group or aa bond or chemical linker group chemical linker group represented represented bybythe theformula formula-(A¹), -(AL)q-, wherein wherein A isAa is a chemical chemical moiety moiety and q and q

is is an integer from an integer 1-100, and from 1-100, andwherein wherein L covalently L is is covalently bound bound to PTM to the the and PTMthe and ULM,the andULM, and

47 provides for for sufficient sufficient binding binding of of the the PTM to the the protein protein target target and the ULM ULM to to anan E3E3 ubiquitin 2023248067 10 Oct 2023 provides PTM to and the ubiquitin ligase to result ligase to result in in target target protein proteinubiquitination. ubiquitination.

[0091]

[0091] In certain embodiments, In certain the linker embodiments, group the linker is -(AL)q-, is -(A¹), group wherein wherein

-(AL)q- isis aa group -(A¹)- group which whichisisconnected connectedtotoatatleast least one oneofofa aULM ULM moiety, moiety, a PTM a PTM moiety, moiety, or a or a combination thereof; combination thereof;

q of q of the the linker linkerisis an aninteger integergreater greaterthan than or or equal equal to to 1; 1; each ALisis independently each AL independentlyselected selectedfrom fromthethegroup consisting group of of consisting a bond, CR¹¹R¹, a bond, CRLRLO, S, 2 SO, , O, S, SO, S02, NR¹, SO, SO 2NRL3 ,SONR¹³, NRL', SONR¹³, SONRL3 , CONR¹³, NRL3 CONRL 4 ,NR¹³SONR¹, CONRL', NRL³CONR¹, NRLSO 2NRL 4CO, , CO, CR¹¹=CR¹²,2 , C=C, CRLl=CRL SiRLlRL 2 ,P(O)R¹¹, C-C,SiR¹¹R¹², P(O)RL, P(O)OR¹¹, P(O)OR, NR¹³C(=NCN)NR¹, NRC(=NCN)NRL 4 , NR 3 C(=NCN), NR¹³C(=NCN),

NR¹³C(=CNO)NR¹, NRLC(=CN0 2 )NRL 4 C3-11cycloalkyl optionally optionally , C3-icycloalkyl substituted with0-6 substitutedwith R and/or 0-6RL¹ RL 2 and/orR¹² groups, C5 . 13 spirocycloalkyl groups, C5-13 spirocycloalkyl optionally substituted with optionally substituted 0-9 R¹¹ with 0-9 RL and/or RL 2groups, and/orR¹² groups,C-C3 iiheterocyclyl optionally 11heterocyclyl optionally substituted substituted with 0-6 RL¹ with 0-6 RLland/or and/or R¹² RL 2 groups, groups, C5-13 C5- 13

spiroheterocycloalkyl optionally spiroheterocycloalkyl optionallysubstituted substitutedwith with 0-8 0-8 R RL¹ and/orand/or RL 2 aryl R¹² groups, groups, aryl optionally substituted with optionally substituted 0-6 R¹¹ with 0-6 R and/or RL 2groups, and/orR¹² groups,heteroaryl heteroaryloptionally optionallysubstituted substituted 2 2 independently are optionally with 0-6 with 0-6 RL¹ RL and/or and/orR¹²RLgroups, groups, where where RL R¹², RL¹ or or RL each, each independently are optionally linked to other linked to othergroups groups to form to form cycloalkyl cycloalkyl and/or and/or heterocyclyl heterocyclyl moiety, optionally moiety, optionally

substituted substituted with with 0-4 0-4 R RLLgroups; and groups; and

RL, RL2 , R¹, RL¹, R¹², RL 3,R¹RLand 4 and RLare, R¹ are, each each independently, H, halo, independently, C-alkyl, H, halo, OC1-alkyl, Ci-8alkyl, SC1-alkyl, OCi-8alkyl, SCi-8alkyl, NHCi-8alkyl,N(C-alkyl), NHC-alkyl, N(Ci-8alkyl)2, C3-lcycloalkyl, C3-11cycloalkyl, aryl, aryl, heteroaryl, heteroaryl, C3.nheterocyclyl,OC1-OC1 C3-1heterocyclyl,

8cycloalkyl, SCi-8cycloalkyl, scycloalkyl, SC-cycloalkyl, NHCi-8cycloalkyl, N(Ci-8cycloalkyl)2, NHC-cycloalkyl, N(C-cycloalkyl), N(Ci-8cycloalkyl)(C N(C-cycloalkyl)(C-

8alkyl), OH, alkyl), OH, NH NH,2 ,SH, SH, SOC-alkyl, SO2Ci-8alkyl, P(O)(OC-8alkyl)(C-8alkyl), P(O)(OC-alkyl)(Calkyl), P(O)(OCi-8alkyl)2, P(O)(OC-alkyl), CC-C1- CC-C 1

. 8alkyl, CCH, alkyl, CH=CH(Ci-8alkyl), C(C-alkyl)=CH(C-alkyl), CCH, CH=CH(C-alkyl), C(Ci-8alkyl)=CH(Ci-8alkyl), C(C-alkyl)=C(C-alkyl), C(Ci-8alkyl)=C(Ci-8alkyl)2, Si(OH) 3, Si(C-alkyl), Si(OH), Si(Ci-8alkyl)3, Si(OH)(C-alkyl), Si(OH)(Ci-8alkyl)2,COC-alkyl, COCi-8alkyl, COH,CO 2H, halogen, halogen, CN,CHF, CN, CF, CF 3, CHF2 , CH 2F, NO, CHF, NO 2SF, , SF5SONHC-alkyl, , SO 2NHC 1.8alkyl, SO 2N(Ci-8alkyl)2, SON(C-alkyl), SONHC 1 .8alkyl, SONHC-alkyl, SON(Ci-8alkyl)2, SON(C-alkyl), CONHCi-8alkyl, CON(Ci-8alkyl)2, CONHC-alkyl, CON(C-alkyl), N(C-8alkyl)CONH(C-8alkyl), N(C-alky1)CONH(C.8alkyl), N(Ci-8alkyl)CON(C1 N(C-alkyl)CON(C. 8alkyl)2, NHCONH(Ci-8alkyl), alkyl), NHCONH(C-salkyl), NHCON(Ci-8alkyl)2, NHCON(C-salkyl), NHCONH 2 , N(Ci-8alkyl)SO2NH(C1 NHCONH, N(C-alkyl)SONH(C. 8alkyl), N(Ci-8alkyl) salkyl), N(C1-alkyl)SO 2SON(C-salkyl), N(Ci-8alkyl)2, NH NH SOSONH(C-alkyl), 2NH(Ci-8alkyl), NH SO 2N(Ci-8alkyl)2, NH NH SON(C-alkyl), NH SO 2NH 2 SO2NH. .

[0092] In certain

[0092] In certain embodiments, embodiments, q of q of the the linker linker is an is an integer integer greater greater than than or or equal equal to 0. to In 0. In certain embodiments, certain embodiments, q is qanisinteger an integer greater greater than than or or to equal equal 1. to 1.

48

2, ALq than 2, A is isa agroup groupwhich is is which 2023248067 10 2023

[0093]

[0093] In In certain certainembodiments, e.g., embodiments, whereq qisis greater e.g., where greater than connected connected to to aaULM or ULM' ULM or moiety(such ULM' moiety (such as as CLM CLMororCLM'), CLM'),and andALALandand AL AL are connected connected qare

Oct via structural units via structural unitsofofthethelinker linker (L). (L).

[0094]

[0094] In certain embodiments, In certain e.g., embodiments, whereq qofofthe e.g.,where the linker 2, AL is 2, linker is is aa group A qis whichisis group which connected to AL connected to ALand 1and to a ULM or a ULM' moiety (such as CLM or CLM'). to a ULM or a ULM' moiety (such as CLM or CLM').

In certain

[0095] In certain

[0095] e.g., e.g., embodiments, embodiments, where where q of q of the the linker linker is 1, the the structure is 1,structure the linker of theoflinker

group L is is -ALI-, and AL -AL-, and AL1isis aa group group which is connected which is connected to toa aULM or ULM' ULM or moiety(such ULM' moiety (such as as CLM CLM ororCLM') CLM')and anda aPTM PTM moiety. moiety.

In certain

[0096] In certain

[0096] embodiments, embodiments, the linker the linker (L) comprises group represented a group arepresented (L) comprises by a by a general general structure selectedfrom structure selected from thethe group group consisting consisting of: of: -NR(CH2)n-(lower -NR(CH)-(lower alkyl)-, -NR(CH)-(lower alkyl)-, -NR(CH2)n-(lower alkoxyl)-,-NR(CH)-(lower alkoxyl)-, -NR(CH2)n-(lower alkoxyl)-OCH2-, alkoxyl)-OCH-,

-NR(CH2)n-(lower -NR(CH)-(lower alkoxyl)-(lower alkyl)-OCH-, alkoxyl)-(lower alkyl)-OCH2-,-NR(CH)-(cycloalkyl)-(lower -NR(CH2)n-(cycloalkyl)-(loweralkyl)- alkyl) OCH 2 -, -NR(CH)-(hetero OCH-, -NR(CH2)n-(heterocycloalkyl)-, cycloalkyl)-, -NR(CHCHO)-(lower -NR(CH2CH20)n-(lower alkyl)-O-CH2-, alkyl)-O-CH-, - cycloalkyl)-O-CH2-, NR(CH2CH20)n-(hetero cycloalkyl)-O-CH-, NR(CHCHO)-(hetero -NR(CH2CH20)n-Aryl-O-CH2-, -NR(CHCHO)-Aryl-O-CH, - NR(CH2CH20)n-(hetero NR(CHCHO)-(hetero aryl)-O-CH2-,-NR(CH2CH20)n-(cyclo aryl)-O-CH-, alkyl)-O-(hetero -NR(CHCHO)-(cyclo alkyl)-O-(hetero aryl)-O-aryl)-O CH 2 -, -NR(CHCHO)-(cyclo CH-, -NR(CH2CH20)n-(cyclo alkyl)-O-Aryl-O-CH2-,-NR(CHCHO)-(lower alkyl)-O-Aryl-O-CH-, -NR(CH2CH20)n-(lower alkyl) alkyl)- NH-Aryl-O-CH2-, -NR(CHCHO)-(lower NH-Aryl-O-CH-, -NR(CH2CH20)n-(loweralkyl)-O-Aryl-CH, alkyl)-O-Aryl-CH2, -NR(CHCHO)n- -NR(CH 2CH 20)n cycloalkyl-O-Aryl-, cycloalkyl-O-Aryl-, -NR(CH2CH20)n-cycloalkyl-O-(heteroaryl)l-, -NR(CHCHO)-cycloalkyl-O-(heteroaryl)l-,-NR(CH 2CH2)n -NR(CHCH)n- (cycloalkyl)-O-(heterocycle)-CH2, (cycloalkyl)-O-(heterocycle)-CH, -NR(CH2CH2)n-(heterocycle)-(heterocycle)-CH2, NR(CHCH)-(heterocycle)-(heterocycle)-CH, -

N(R1R2)-(heterocycle)-CH2; where N(R1R2)-(heterocycle)-CH; where n of n of the the linker linkercan canbebe0 to 0 to10;10; R ofthe R of thelinker linkercan canbebe H, H, lower lower alkyl; alkyl;

RI and R1 andR2 R2ofofthe the linker linker can can form formaa ring ring with with the the connecting N. connecting N.

[0097]

[0097] In certain embodiments, In certain the ALthe embodiments, group AL is represented group by a general is represented structure structure by a general selected selected

from thegroup from the group consisting consisting of: of:

-N(R)-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-, -N(R)-(CH2)-O(CH2)-O(CH2)-O(CH2))-O(CH2)q-O(CH2)-OCH2-, -0-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-, -O-(CH2)-O(CH2)-O(CH2)-O(CH2)p-O(CH2)q-O(CH2)-OCH2-, -0-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-O-; -O-(CH2)m-O(CH2)-O(CH2)-O(CH2)p-O(CH2)q-O(CH2)r-O+; -N(R)-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-O-; -N(R)-(CH2)-O(CH2)-O(CH2)-O(CH2)p+O(CH2)-O(CH2)-O-; -(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-O-; -(CH2)O(CH2)-O(CH2)-O(CH2)-O(CH2)q-O(CH2)+-O-; -(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-; -(CH2)-O(CH2)-O(CH2)-O(CH2)-O(CH2)q-O(CH2)-OCH2-;

49

2023248067 10 2023

-(0H 2 ),O (0H -(CH)O (CH)- N N- N(CH)O 2 )r7 N 00(OH 2)p - (0H 2 ).(CH)- ;

Oct -1CH 2 )m-N \/N-(CH 2 )n-NH

{1(CH 2 )m-N \-/N-(CH 2 )n-O7

-:-(CH 2 )mO(CH 2 )n-N N-(CH 2 )o-NH

't +(CH)O(CH) N N-(CH)-O 4(CH2 )mO(CH 2 )nN&N-(CH 2 )o-NH

-(CH)mO(CH) N X N-(CH)-NH

±(CH 2 )mO(CH 2 )nNX\N-CH2 ).-O' --I-(CH)O(CH) / N N-(CH)-O

++- (CH 2 )m-:, (CH);- -:-N -i-N 0 O N N -'-NO -,-N 0 O ; N N ;

(CH);- NICHH2)m 4

N (CH) _!_ -:~(HO -+- ; (CH) ; N N-(CH); N IN'.,;-N

O ; (CH) ;

o

N N 0__ 0 N N

o o -N / \N-// /N F \N -N

N N 0_/N N N N N

o o

50

N \/N-CH 2)mO(CH 2),,O(CH 2)pO(CH 2)0, ;

H N HN N / O(CH 2 )mO(CH 2 )nO(CH 2)pO(CH 2)qOCH O(CH)O(CH)O(CH)pO(CH)OCH. 2

X ;

NH 2023248067 H/ O(CH 2 )mO(CH 2)nO(CH 2 )pO(CH 2 )qOCH 2 O(CH)O(CH)O(CH)pO(CH)qOCH ;

NH NH -4- NO(CH 2)mO(CH 2)nO(CH 2)pO(CH 2)qOCH 2 1111. O(CH)O(CH)O(CH)pO(CHqOCH ;

NH H2NH)CH \/ O(CH 2 )mO(CH 2)nO(CH 2)pO(CH 2)qOCH 2 O(CH)O(CH)O(CH)O(CH)qOCH ;

_ NH -:-NH \/ O(CH 2 )mO(CH 2 )nOCH 2 O(CH)O(CH)OCH ;

O(CH 2 )mO(CH 2 )nOCH 2 O(CH)O(CH)OCH HN X X ;and ; and

N ZI N \ H H N-(CH 2 )mOCH 2 N (CH)OCH N N ; wherein ; wherein

m, n, m, n, o,o, p, p, q, andrrofofthe q, and thelinker linkerare areindependently independently 4,6;5,7,6;8,7, 9,8, 10, 0, 1,0,2,1,3,2,4,3,5, 9, 10, 11, 11, 12, 14, 12, 13, 13, 14, 15, 16, 17, 15, 16, 17, 18, 18, 19, 19,oror20; 20; whenthe when thenumber numberis iszero, zero, there there is is no no N-0 or 0-0 N-O or 0-0bond bond R of R ofthe thelinker linkerisisH,H,methyl methyland and ethyl; ethyl;

Xofofthe X thelinker linkerisisHHandand F F

''NN IZ \ H / / N(CH 2 )mOCH 2 H N-(CH)OCH N where where m m of of thethe linker linker can can be3,2,4,3,5;4, 5; be 2,

51

IZ IZ IZ H H IZ IZ H IZ IZ N N H IZ IZ H HN N NN' O HN

N N H- H-: o ZI

HN

o 0HIl 0

o o IZ

o

O o 0 :

......!! IIIIIIIII. N N N N N N

H2

IZ H IZ /N IZ N IZ H H H H IZ N IZ N' H H IZ IZ IZ H H H N H

÷/-o IZ

H /N TNNo IZ

H 0 o 0,0 IZ IZ S/ S 0 H H , oN H, IZ

H H H IZ IZ IZ H

N 0 H, HHH o IZ N IZ O

o IZ HN Ci/ HN H H HH

IZ N N IZ H H N H IZ 'N N

N H ZI N IZ IZ H H H -H N ' NJ'~ H ' N"'I N' "~1 o O H '

N IZ N N NH H IZ N H 'N H 0 /N -NH 0H ~/, ZI H IZ

NH O N f

o HN N o N 53H O HN N N n

N IZ H X

in

O~~O

OH Ho 0

0

0 - 0 O O ~ /

a a O

/ ~LK~ O H O Kz~o~j< ZI N / 0/

H IZ 0 0 0 N N ~O N

\~-~--~---a a/ a / a 5 a O 9 a

54 SA

rai, ronal, marry's resort regles somes N N

0 0 0 2023248067

N N ; swag ; Onaly 0 nN 0

00

Oraly N Ronal ; Onaly N 0

0 0

Onaly Onaly ; Anal N NN N NN0

0N-A o - -,,

analy. Onaly. aor N'p N

N N-N 0 0 -oN N N ;

N N N N NN N N ; will

O o N /5 N N N N N in

N ; HO HO

0O N 0 O N 25 s N }{ N N ; O ZNN ,,,

O 3 O 3 3'm /\NN N-- N N-0 N -I-Nr in N N-- N in N -- N N N N ; ; ;

viv in N N N-m N N N N-

56

N'I' ZI IZ IZ N N "N N 1111.

' i H H H O x X

IZ H IZ H N ZI N N N H H H /~ XH, F X=H,F X X H ZI H ZI H, H N N IZ O N7N N H ~ N N o N HH ZI H IZ N N N N IZ 0 H N 'H7 O N N H H ZI H IZ N IZ N N H H O 0 ~ IZ IZ IZ N H N 11111

N IIII

H H H N o H~ 'H H

IZ IZ IZ N N N N H N7/ H N xX xX IZ 1110 H IZ IZ

H N // N 1110

H H N N X ZI H IZ N HN/ N IZ N H N N H N N N

N= xX x X NN -C- I HN HN NN N N

57

Oct 2023 -:-NH - ~NH N NH NH NI N- N '--NH N N NH 0 - 0 N N N - o N O \/ 0 O 0 o -:-NH 0 -NHNH I - -NH NH o NH N N N 0-N- N --/-\ N N N 2023248067 10 N o U 0 O 0 O

-'-NH NH 0 -'-NH NH I-: - -NH NH 0 o o o

NH O ZI H IZ N N A 0 \ H -\ \ \

o ZI H o N N H ZI N IZ N Z~is 1110

S NHIH IZ H o H N N

iC I~u /\ H 0 'Nr0 N X HN HN HN o x X=H, F N X X=H,F X X

HN O N \ / '-N HN -K~o / _ _N HN m-oi-oO x./ N N N

HN HN HN N O IZ IZ millo N-O N N

N N N O N N N N N -N~ a NO N N

N 0/NC-\ N N L O O NN

58

HN O O HN HN N N N O N

HN HN HN~K)N HN x N X H*- / ' - O IZ IZ HN H H Z IIIII

H x X x X "00 HNPKI o ' /N~ ZI / "N IZ HN N N H H H

. H x X 0'

H ZI H N H ZI H H ZI N N 00' 0 0'1 O

HN o 0 w HN H\ 0- HN-0 N ~ N HN o HN N

HN N$\H

HN N HN N N HN O

N N N IZ N IZ N N H- H H \/z N 0~ H O STATE

-+- N -'-N N NCN--' -:-N N-~ 'N NN-" N N N -|-N N N

59

2023248067 10 2023

-N-N-N' N N N -|-NN N N N N -- N NXNN N N THE Oct

N N N N HO Ho HO Ho -:-NN N N _ N -|-N N-/ N -N N -- N N N 0-' N

0-~~0 o N N N N N NNN N HN N 0 O 1

ON/OI -|-N N NN HN N N N N N N 0-1 N o

O |-N N ,-N HN N ' N O HN N

N NN N HN N HN N N HN N N N

NH O OH HN N HN N\ 'N N\ O HN N HH N HN~~ 60 N ., O N ZI H N HN N o / F F FF

where each where each nandrmnof n and the linker m of the linker can independently can independently be3,0,1, be 0, 1, 2, 6. 4, 5,6. 2,3, 4, 5,

100981

[0098] In In any any aspect aspect or orembodiment describedherein, embodiment described herein,the theAL group isisselected A'group selected from fromthe the group consisting of:. group consisting of:

60

N N N' N N N: N N N N N N N-.N

N N N N N N o... N N N N 0_0

N N N N.., O-/- N N=N N N," N

N N N 0 -N N N N N p N. NNo'*

N N N N N NN N N to N m~- N " "-" N

N \\00-'NN.,' N N w N N N N w N 'I

N N N N N N N N N N N.'

N \,\N N N N N N N N N N NCI 0 N -NC

H HH H- H

/N~f N H

HN HN 61 I9

E o o

IZ ZI NN HN IZ

0- 0

N ZI N HN HN ZI N H ZI

O o ZI

H HIH

IZ ZI Ni HN

H

0 N ZI ZI ZI

NN"

H 0; HH ZI ZI ZI

O ZI 0 N ZI N

N 0 62 IZ IZ HN

Oct 2023

H ,_ 0 stonaly snagal mayor subraral HH N0 N 0 H F 0

, 0 /I,,, N Ny 1 NN N ,, NN I 2023248067 10

smol, may may may H

.,0 N , H

/ o0 0 0 0 0

0N~ o

0IC N 0 -,fA maraly snaggs , ,o ; olI O0

00

small final magnis snages, H IF 0 0N

0 0 00

snargly snangly /~N msais magal, H- H H 0 NN N Y N

0 0 0

ingal, ~ mggal ingoris And N N N N H >I.: ,

H i HH N -F H

0 0~ 0 0 N ."O N

moors. 0 O H -- H0 H H 0

0

sports smoul H N

F o? H N - V - - -x 0 -/0 Na o'S-o

HF

0 0 0

H N /F magal sport q 4NO0

H No, '

H' N,

0

~ q 0 0,' 0S N 0r>f OI

H [H H NN ~o "F"Or" regal, 0 F

0 0K~ 00 0\W~ N NN

subses goods HKNO F

0 N N. ~K~H F F

H0 00

F

0~ years Hair Haus 0 HE

mg wagal, sports more F H N F IS~ 0

63

ZI N H N ONH O s N O ZI F H H H FF F F FF

O o o /I NNIII o N..Q- ."NI"Nh! N N N r ZI ZI N ZI N H H H H H and and N H

wherein each wherein each m and m and n is n is independently independently selectedselected from from 0, 1, 1, 2,or3, 6.4, 5 , or 6. 0, 4,5, 2, 3, 2023248067

[0099] In any

[0099] In aspect or embodiment any aspect described or embodiment herein, herein, described AL group ALisgroup selected from thefrom is selected group the group consisting of: consisting of:

O ZI O ZI N O N N H H ZI O ZI N N N H H HH

N ZI N O0 N0 ZI O N H H

N H O N H H H O I ONzzN ZI N N ZI N H H

N N %O o N N N N O O N No

o

N N N N ,O N N N N O N N

o o

64

NKN N N .1NNN

' N N III H IZ

,,-0 Ni N 1.0 N N IZ N H NN,,N, N NN N, IZ H N~, N N")

N N N ZI NN%~

N N ZI IZ

N N N N oN,<'o(0\ F F FF FEF FF FEF F F

N N N N N N ZI ZI F F H FEF H; FF FF N N

ZI N N FF FE IZ H1 . FNE'1 F F lizN''N' FF N FF N FE H N N ZI

FF F F

65 $9

Oct 2023

N O N N 0 N N NZ~ N IZ N IZ N FEF H H H H 2023248067 10 FF

N H HH IZ ZI

H 0," NN IZ N HH ZI 0o H oH N OH OH ; OH OH 0 O "' O"~"~"O N" IZ N OHH OH ZI HH H H OH OH *. O N

OH0 OH IZ N H O O H. IZ

O H

H ZI H N -',' H ZI H o O 0 O IZ

H;

0 0 o o IZ IZ

H H ;~ H H

0 o IZ N H

66

o IZ IZ 1-0 N H H

IZ H H

o N N o N N m n n m mm

N N

m n o N m n

n N N % N n n o O m

-:-NN N- N -:-NN NN N N 0 m m

N\-----O

-- f" n N N N N N m

n m n n m m

0 (%%O\\ o / N N N N-0(%O% -:-N N N-V N f * m nl n 0 m o o

67

m010 HN tax N n n n

mm Y

% n n n n n

n N 0 o

0% N' ot

' m

m n N n

N N XIX o

N N N m o N N m NH N N m N - N-- /) n - S

N + N m O N m NH N m 00 0

o N o NH (/) I-H

N N N N n n O o m m

o

N IZ N NH m B n n

N-'68

N N N N m m N

Nm - N ?~ >NN m1- - N m 3/2/20

N + N N + N m N m m n

m N

N + N + N

m + N N r r~N N

N N

m WE N N N N.

N N

m m mm m WE N N N WE N N N N N

m m

N6N

H ZI H HN ZI N N

2023248067 10 Oct m NNN

ZI pn 0 N N n p n N N IZ pn N N

O\ N N-\ n n p - n OV NN

n n

N N N N n N n N

N N

m m 4 m m

N N N m m

N N

N N Na o m m

70

N F FF N N N ,,.o0 N N N

n m m m n m N 0OH N OH N ZI

N N m N m x n N m n m CF 3 N N OH N CF 0 OH N N N N N O 2< '0n NN n n N m n m N o N O N '\ N N o m xin m N N NZ N-:-X_ N N n O N N N O. NN Xl~ N N N N

NNN m n n N N N N mm N o n N N N n 00 fl m N N N N -mNIO n OH m o NN D N n NJ N m n N /N- Nk n N O-N N n m

n ~NN N F m

N N ~N m N N p m n ZI

n~ ~ m n N N N,4"

N N H ~Nj n N n N N Ii n0l, IZ

N N you N'N' N" N F N m n H r N N N N m N 0n m m N

n m F7

2023248067 10 2023

N N N n NN: N Oct N N m IN/ N m N

N IH n

m IZ N H n NH NH2 N N

o o m N N n O m o NN NN o o m n 0, 0 ---0

n N N

o o N N m HO N N p O N n N HO n m m o Po~W>) p n m HO N N N mN N n n m 00

N N

N N N 7HO m n

o

H

Z Z h N0 HN

IZ NX NN

N N N N 'N/ N N m N N N N N w

N w N E 3'3' N w N m N/ m N/ N H N E 3'3' E N N N m u ZZN 3 '-NN N NC u NON/~ N N N N w N N w 0 O S m N N w 3'3" w N -/~N N N

N NN N N w N N HN N N u m0

73 EL

N NNI n N N N N N u N''~ C:N N N

N N NN N N N N,, N N u N. I N N\-~'N~.-NN ~-,I N -, N - 0/ u N N N N N N w N N N

N N' N N N N N-' N 0-/- NN N IN N

N N NNN N NN N N mm w w

- ~~\ b/-\N Na -N"' N N N N N N N N

N N N N //o -:N N

N N NNI N N N N % m N-N N N N N N N N El- u N N u

0-÷ -:-N N N N u N N u

CFF N N N N E N N

74

O O O O 00 0 o 0-11 p m n

o m o n O 0 o

n o m 0 H ZI H QQ0 o o o /'1 0 o N 11

m 4n / m% m n m ZI H O O o o N

0~~ 0 0 N N m m n ZI

o O O O N HH n o p m ZI H O O O I'-f

mm n m n n oo p

000 O N 0 0 n O no m mn m n 0o m o o o O o IH

n o p q m o O o o

nn n o p m ZI

oo oo o0 ,, o0% mn n 0 q m m ZI

O O o O

o0 m n m 0 n ZI 'I H O o o o n o p q m m n 075 ZI

o O o

n o p m

o N O O

n o p m

N 11 N rlN N n N

N

N". to 0 ~ N

to N

N- m N

+ N0,

0'l

0_ N

N N+0+0+ + m N _

M m

N \ 0N

NN

m m

0,1

N m 76N

Any N N

N N. N +0-0+nx N N N / NN N

N

o~ N' N

N N

+0-0-0+ 0

_ - N - :- _ NN

ro o

0 m

77 X

things

H) H2

IZ m N m N N N m n

O N NN N-- o 6H) NH-- NH oo n m m 0 m m4n N N m N N n m .....!!! IIIIIIIII.

N N N N mNO Ni -:-o\

O NN N N N N n m o

N /--N N N-I-- - : N N N N o 00 o

-/ X/\ N N mNKXV/ 0 n N N -- :-o N N o O n N

N mNI-78 N

N N N n n o m

N N m N N n n o

ON nN 2023248067

N

X N N N N n 00

o N 00N N N N N n m m n m o o N. NH/ NH % -NH '- 0-/&N- / /

......!!

N 0I N n HN m N N m o o n N m

X NH X NH

m N n

O N n X ZI

o m N n

m

X mN N N(~N ~79

N )emN N 0-k0 m N \N- N0 N N

-N N N-

N N n N Nm N - + m m N O

X N N N n m m N N- N N-4&

m m N -/&NN n X~ o -

mO 0m N 0 o

04 - 0 n N N Nt N N 0 -,-- m

N N N N

800

0N~NN '0 'A'

N

-' white o m

m o

m m

N " -N -N 0

0 N

n 0

N 0

0 0 0 m m m N 0 n ' 0 N 0 ' -' p.. 0 N N 0' n m -'N 0 0 N N N N,' ifi m 0x m ' 0 ---/--

\A 0~~0O n m

N N 0 N

0

N,

N ~ ['N0

N ' N.0 N,' -, N A, N 0 0

n

N, ~ 0 N 0 0 N,' ("'N

0 ~ 0 A"~NN,,A' 0 0 0 N

~',~0 0 /

'N N 0 o o N II m 0 /'~'-0

0 0

N' 0 0 , 0

XXXXXXX --/-- >0 n m N H

81

N

0 N

n nn

0'

0",O

0 N0 to they n0

0 n 0 N

name 0 N

' 0 0-N

0- I Ir

This NN 0 NN 0 0=

think ,,- 00o-4 0 NN

00

00

0 0

82

NC

N 0/ to 00 00 -\Y N

N --/--

N 0°3

Not

00

00N

ot 0' 0

ui

N N 00 ON NC

0 NN It n 00

0 ---. 0CN' FEE I N n

/ 0 0 N --- N 3 0 N o

N N O

0 N

ON NC '00 N

83 E8

0' ',0 0 nF F

F FF

*Dratcopt F.

00 F

2023248067

"0

N CF N CF

N..,, N/ N F3

the N F3C 00

N 0 0 N

any F FC

0

0

N

N 84

CF 3 CF

m m m m n 0n n

OH

*0

n

OH o~tio

0

OH5 n

m m

N N

+

00

N

00 00

N l-5'

2023248067

NC

N 00 N

N ~ , oxl

0 NN

N

'0 N

F3C

NNN

F3 C N

0 0 ,0N N,>5 N F3C N

00

N. N o N NO N0N N N.

NC NN

05 HO NN

N N.

N 0 N N

N ,,>~ 0 --- O N CN

0 C N N

N. N N N

86

N

00

04 .N. N 0> '00 CF

CF 3

00

0 N 0

CF

0'

141 " N

'00

CF

"'O0

0

-- N

00

oxo '00

'0O.

N Nc

NC

o' N N 0>

87

Oct 2023

N N N

N N m NJ 0 2023248067 10 FC N N N 0 N NN

N N0

N

N +0+1.0+ T I~~~0

N N- N

N m 00 N n

-11 1N N m0N

* 00

NJN N

- 0 N N 00 N

N O

0 o0 m 0 N 0N

0 00 N

N 0 N

N HO

.0 N 0

N 0 \10N N 0 O HOJ

N

N 88'

N N NN r, r 4-,m Nj N ,N N 0O 0 %%

m nn p p

N N N rl* N r r j 0o N N N %N N 0 q N m n n p p

N N N N N N

m n

\\, N0 N

mm

i NH2

N N o o n m n

X + N

o m n

N N

N N N 8N

m n p

H IZ

N NN ZI N N N H m

N o o O N 0 N 0 IZ

HH ZI N N m u N N 00 o o o H IZ

N N N 0-'S N m m 00 N o o H IZ

N y N N N N N o o

N N N N HN N N N N

'5, ; 0

N N N N

09 ZI N N

N N N N O ON O N N N

N N ON ONJ N Y- O O 0O., O O CF 3 N; N O' CF O/

N ,wherein , eachm, m, wherein each n, o, n, o, p, p, q, q, andand r isr is independently independently 0, 1, 0, 2, 1, 2, 3, 4, 5, 5, 6, 6, 7, 8, 9,10,11,12,13,14,15,16,17,18,19, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, oror 20. 20.

[0100]

[0100] In any In any aspect or embodiment aspect or embodiment described describedherein, herein,the groupisisselected theALALgroup fromthe selected from the group consisting group consisting of:of:

"A -- 0 0- ;

O -0 0 0 0 0 a a 0 0--1 ;

O O O O-\ O- /O\ O-- N. O-\ O- O- O -~NH NH -0 HNO HN - o- HN ---. 0- 0 ' 0 0__ o HN O o O O ;

NH; O HN 0 0 0 0 o NH O O

; ; ; 0 0-/

-- 0 0 0-- - ; ;

-- - -- -

0 ;

0 0

91

0 0--*-\ 0 0

0 0-

0- 00 ~ \

, O O _ -- C NI,

-0-0-- - - 0 0 -\/ 0 0-/ a/ N

N N ó -0 N -

N N N

a o-- N-, 0--- -- N0 -\ N N

N N N N

0 0-- 0N N 02 N N

N N N N N N N - NNN N N N-N

0

N N N N N-/- N N- C N-- N -- N

~-- 0 00-- -

0 N N N 0

N N

- 00 N

N 0' 0 O N N N N HN

-0 NH N- NH NH HN N---o 0 00 O 0

/ - J-O HN--- NH _/0 -0

O HN 0 0 0HN 00-'

NH 93a HN

E6

2023248067 10 Oct 2023

o -- /\O -0 OHN-- O HN ;

0 HN 0 o 00N NH O O o O 00 0 0 HN o NH 0 O NH 0. 0 O HN -- O o ;

- p -N-- 0- - N 00 O O O NH HN O o HN O ; ; ;

\/ 0HNN----- H/ NOH- O O O O O HN ; O HN ;

o O NH O O NH 0 ; O ;

00 o NH NH O ; O ;

N // or

% O o o O 00 _/ NH 0 0 O NH O HN0 O O ; ;

0 /N

0 HN 0 NH NH O ; O ; ;

0 NHN0 HN- o o NH NH O 00 HN __H -- HN--- N O O o o o O NH4 HN HN HN ; ; ;

O O O HN HN ;

0 0 2023248067 10 Oct 2023

NH N NH -NH NH 0 0 0 __\/0 o 0O o 0 O

0 0 O NH _ NH O O O 0 0 0 HN-- O HN

O 0 0 O O -- o \/ 0 0 OOOHN HN -----\0 O/- -- 0HN-- O HN 0 O 0 o 0_C __/- HN--- HN -NIH NH N N O N O HN O 0 / 0 % 0 HN 0

O N --- N NH-- NH N N N NH 0 O 0 O ;

O O HN 00 NH N N O -- 0 N0 N 0-N 0 HO 00 N O HN N NH

0 0 0 N N HN o HN 0 N O

f~N (N o 00 00 N NH HN-- o N N 0 O N00

-N NH -'N N N N 0 O N N NH N o O NH NN O o

N v-95 N N NH N NH O o

Oct 2023

N N N N O / N N NH N NH 0 0No N HN, HN O O O -- C N N-__ N-C N NH NH ;N/H O 0 HN -- HN---0 o 2023248067 10

NH NH NH O O O O 0 0 0 O O o NH NH r-\ N NH HN--HN-; o O HN HN N ,\~N-j N-o N-O ,0 O N-0 N-O O 0-u - U/

/ N-O N-O O ;

O N-O o N-O N- 0 O

O N-O N-0 N-O N-0 O O -0 0 -0 N-O N-O O K 0N-0 N-0 ;

N-O O N-O O

N-C N-O /N-0 N-0 O N-O O

N-0 N N-0 N-O N-O O O ;

N-O N-O N-0 o N- 0 ;

N-O N-O

96

N-O N-O

N-O NNI- - NN N N-0 N N

N N-o N N '- N N

N

NNN1/ O-0\ N N N-o N N N N -0

N /\ \--_ N N -0 N-O 0 N N N

N N N N N N N

N N-O N N-O N N ~ N~jN-0 N-0N~

N N-O N N-O N N

\ -- 0--\ /- 0-\ -J I O

-- O-.-0 o O 0- -- F0 00 i o O O

--o Ór--\ O/-\ a0\ -- \ -- a a- __ -- a -0-a/a--.__a--. --o O O O O

o O --o O 0. O -- a -, -- 0 0-/- -- 0 0 0--.--0\0 0- 0--.

97

2023248067 10 Oct 2023

O o -- 0 0. 0 0--.--0 0 0--.--00-

-- N N/-\N-- N

0 0--_ / -- N N\ /N---- N -- N N\-/N- N 0-; -- N NN 0d -- N N \-/NN N

N _ -N N--- N 0_ O _

-- N N N N "'-N N N 0_ _N NN-- N N O N

N l\ jN N N --- -- N N N'--N N N \-/NN

-- N N -- N N -- N

N\ N -- 0 _N N NN__/ -- N N NA N 0 0-

--I N N/ \N N 0 0-- 1J N -- N \N 0 a--\_O__NN/ O \N_0 N

N-, N--- -- 0N N0 N N -- N N/-\N-\ N N N N N O N

0 -0 HN--- 0 0 o NH NH HN -- 0 O 0__--0 O 0 N HN \.H-0 'i0 0

o 0 0 O /0 0 O -~0 0-\ NH HN -- 00_\-/ NH ---.--0 o0OHN 0 0_ 0

0 H N-- O NH O O_\ 0 0 NH-- HN -0 NH HN / 0 O_0 0 0 o 0. 0 O

O 0 O 0 O 0_ 0 -- 0 / HN __ ` HN --- 0 HN NH NH NH

O o -0 0 NH -- 0 HN 0 O NH --0 NH- O HN HN O O NH 000 O O NH 0-- NH 0 N HN o HN O HN 0_ 0 0 O O

98

O 0 O 0 O NH NH 0 o NH NH NH NH - 0 .-- O 0 O 0 O O 0 0 0 NH O NH O0-- NH .O NH NH NH 0 O 0 o -- O 0 O -- OO

0O NH 00HN O O NH O NH NH -N 2 2023248067

NH 0H O O f-K -- 0 0_ 0__N-.-0 O O __-- O NH -- 0 O'O--; OO'.O O HN -- O O HN O O o O O 0 OO --0 OHN---. O HN OO O0O O HN -- O ;

0 O 0 O 0 HN---. OO O N---. O OO O O HN O O HN ;

o O O 0 0 O O OHNO O 0 HN 0 o HN HN---. ;

00O O O O O 0HNm 0- O \-0 / HN HN HN ;

O O O O HN O HN ;

0 o a o o-- Oa o- a N-- HN N-- N N N N -- 0 O HN--N HNNON O O HN HN O

ao a o aN /- -- N-- NH N H - -- N O NNH ---N N N N ON O -N N N O HN ON---N N NH O 00 O o O O HN -N N NH N N ON-F--j O O a a O O NH -- N --N N N 0 o N -- N N N N O O - O HN

99

000 O HN HN HN o O O N 0-- N -- N --N N- N -0 O HN~; HN N N

0 O O N N~-- J NH NH N N N/\N---' -- N N HN __0 o-0N N N -O NH N 0 O ;

N NN \ N N N -- 0 0--- \s-j - NH--C/ C- NH o NH O O NH 00 O ; O ;

N/ \N/N/ N N N -- N --N N-- N - NH NH ---N N N N NH NH 0C O O ;

N O --N N N -N N ~~N ' 0-N N N N NH 0 NH 0 NN N HN O ; O

N-0 0 N-O N-O N-0 N-O N- N-O O O ;

N-O N-O N-O O

N-O N-O N-O O 0 O 0 N N- N

N-O N-O N-O o O

N-0 N-O N N-O O 0 N-0 N- N-O N-O

0 0 N- N-O N- N-O I / -. I

100

N- 0 N- 0 Oct 2023

N-O N-O O O0 /O

N- 0 N-O N- N-O 0

O 2023248067 10 ; O

N- 0 N-O /\ N.- 0 N " N N-O /- -- NN N -/ N N N- O N 0 O N0 N N o ; ;

N-O N-N N0 N-O ('N N N \N N N N N-O N N-/-N~ N N

N-O N N-O -- N N-O N N O- N O N N s'O N O -N 0 NN N N-O N N N N-O N N

N-O N-O N o N N N O

N-O N N N N N N - NN O N ; and ; and NN)/

[0101]

[0101] In additionalembodiments, In additional embodiments, the linker the linker (L) comprises (L) comprises a structure a structure selected selected from, but from, but

not limited to the structure shown below, where a dashed line indicates the attachment point to not limited to the structure shown below, where a dashed line indicates the attachment point to

the the PTM or ULM PTM or moieties: ULM moieties:

(YL¹) 0-2 WyL10-2 (YL¹) WL1 WL2 WL1 WL2 LL0-2 O / n n / n

or or

wherein: wherein:

2 WL andWL² WL¹ and WLareareeach eachindependently independently absent,a 4-8 absent, a 4-8membered membered ring ring withwith 0-4 0-4 heteroatoms, heteroatoms, optionally optionally substituted substituted with with RQ, Rº, each each RQ is independently R° is independently aa H, halo, OH, H, halo, CN,CF, OH, CN, CFC-C 3 , CI-C6

alkyl (linear, branched, alkyl (linear, branched,optionally optionally substituted),C substituted), 1 -C 6 (linear, C-C alkoxy alkoxy branched, (linear, branched, optionally optionally

101

Oct 2023 substituted), or22R°RQgroups substituted), or groups taken taken together together with with the thethey atom atom arethey are attached attached to, form ato, 4-8form a 4-8

membered membered ringsystem ring system containing containing 0-40-4 heteroatoms; heteroatoms;

Y" is each YL¹ is each independently independently aa bond, bond, C-C C 1-C6 alkyl (linear, branched, optionally substituted) and alkyl (linear, branched, optionally substituted) and

optionally optionally one one or or more more CCatoms atomsare arereplaced replacedwith withO;0;ororC C-C1-Calkoxy (linear, alkoxy (linear, branched, branched, 2023248067 10

optionally substituted); optionally substituted);

n is 0-10; and n is 0-10; and

aa dashed line indicates dashed line indicates the theattachment attachment point point to tothe thePTM or ULM PTM or moieties. ULM moieties.

[0102]

[0102] In additional embodiments, the linker (L) comprises a structure selected from, but In additional embodiments, the linker (L) comprises a structure selected from, but

not limited to the structure shown below, where a dashed line indicates the attachment point to not limited to the structure shown below, where a dashed line indicates the attachment point to

the PTM the or ULM PTM or moieties: ULM moieties:

(R°)- (YL¹) (yLl)0-2 WL 2 O WL2 WLI WL1 QLn QL n

or or

(R°)- (YL¹) (yLl)0-2 L2 WL2 my W WL QL QL W n

wherein: wherein:

are2 are cyclic, heterocyclic,C independently 1-6alkyl WL and WL¹ and WL²WL each each independently absent, absent, aryl, heteroaryl, aryl, heteroaryl, cyclic, heterocyclic, C1-6 alkyl

and optionally one and optionally or more one or more CCatoms atomsare arereplaced replacedwith withO,O,C 1-6alkene C- alkene and and optionally optionally one one

or or more more CCatoms atomsare arereplaced replacedwith withO,O,C 1-6alkyne C- alkyne and and optionally optionally one one or more or more C atoms C atoms are are

replaced with 0, bicyclic, biaryl, biheteroaryl,or biheterocyclic, each optionally replaced with O, bicyclic, biaryl, biheteroaryl,or biheterocyclic, each optionally

substituted substituted with with RQ, R°, each each RQ is independently R° is independently aa H, H, halo, halo, OH, CN,CF, OH, CN, CFhydroxyl, 3 , hydroxyl, nitro,C C nitro,

-=CH, CH,C2-6 C2-6alkenyl, alkenyl, C2-6 C2-6 alkynyl, alkynyl, C-C 6 alkyl CI-Calkyl (linear, branched, (linear, branched,optionally optionallysubstituted), substituted), C 1-Calkoxy C-C 6 alkoxy (linear, branched,optionally (linear,branched, OCi-3alkyl substituted), OC-alkyl optionallysubstituted), (optionally (optionally substituted substituted

by 11 or by or more -F), OH, more -F), OH,NH, NH NRY¹R², CN, 2or 2 R° groups taken together with the atom 2 , NR'Rv , CN, or 2 RQ groups taken together with the atom they are they are attached attached to, to,form form aa4-8 4-8membered ringsystem membered ring systemcontaining containing0-4 0-4heteroatoms; heteroatoms;

102

YL¹ is each Y" is each independently independently aa bond, bond, NRYL¹, NR",O,O,S,s,NRYL², NRYL 2CRYL¹RYL², C=0,C=0, C=S, C=S, SO, SO, SO,C-S02, 2023248067 10 Oct 2023

, CR YL2, C1

alkyl(linear, C 6alkyl C (linear, branched, substituted)and optionally substituted) branched, optionally optionallyone and optionally one or more C ormore atoms are C atoms are replaced with replaced with O; 0; C-C C1 -C6 alkoxy alkoxy (linear,branched, (linear, branched, optionally optionally substituted); substituted);

QL is a 3-6 QL is a 3-6 membered alicyclic or aromatic ring with 0-4 heteroatoms, optionally membered alicyclic or aromatic ring with 0-4 heteroatoms,optionally bridged, bridged, optionally optionally substituted substituted with with 0-6 0-6 RQ, R°, each each RQ R° is independently H, is independently H, C-C 1alkyl .6 alkyl(linear, (linear, branched, optionally branched, optionally substituted substituted by by 11 or or more halo, C more halo, C-1 .6alkoxyl), alkoxyl), oror 22 R° RQgroups groupstaken taken together with the atom together they are atom they are attached to, to,form form aa 3-8 3-8 membered ringsystem membered ring systemcontaining containing 0-2 heteroatoms); heteroatoms);

R¹¹, , RYLeach RYL are 2 areindependently each independently H, alkyl H, OH, C- OH, C(linear, 1 .6 alkyl (linear, branched, optionally substituted branched, optionally substituted

by 11 orormore by morehalo, C- C halo, alkoxyl), 1 .6 or R¹, R² together 2 with the atom they are attached to, alkoxyl), or R, R together with the atom they are attached to, form form aa 3-8 3-8 membered membered ringsystem ring system containing containing 0-20-2 heteroatoms); heteroatoms);

n is n is 0-10; and 0-10; and

a dashed line indicates dashed line indicates the theattachment attachment point point to tothe thePTM or ULM PTM or moieties. ULM moieties.

[0103] InInadditional

[0103] embodiments,thethe additional embodiments, linker linker group group is optionally is optionally substituted substituted (poly)ethyleneglycol having (poly)ethyleneglycol havingbetween between1 and 1 and about about 100100 ethylene ethylene glycol glycol units,between units, between about about 1 and 1 and

about 50 ethylene about 50 ethylene glycol glycolunits, units, between between1 1and and about about 25 25 ethylene ethylene glycol glycol units, units, between between about about 1 1 and 10 and 10 ethylene ethylene glycol glycolunits, units, between between1 1and andabout about 8 ethylene 8 ethylene glycol glycol unitsandand units 1 and 1 and 6 ethylene 6 ethylene

glycol units, between glycol units, between2 and 2 and 4 ethylene 4 ethylene glycol glycol units,or units,or optionally optionally substituted substituted alkyl groups alkyl groups

interdispersed interdispersed with optionally substituted, with optionally substituted, O, 0, N, N, S, S, P P or Si Si atoms. In In certain certain embodiments, embodiments,thethe

linker is substituted linker is withanan substituted with aryl,phenyl, aryl, phenyl, benzyl, benzyl, alkyl, alkyl, alkylene, alkylene, or heterocycle or heterocycle group. group. In certainIn certain

embodiments, thelinker embodiments, the linkermay maybebeasymmetric asymmetric or or symmetrical. symmetrical.

[0104] In any

[0104] In of anythe of embodiments the embodiments of the of the compounds compounds describeddescribed herein, herein, the thegroup linker linkermay group may be any be any suitable suitable moiety moietyasasdescribed describedherein. herein.In In oneone embodiment, embodiment, the linker the linker is a substituted is a substituted or or unsubstituted polyethylene unsubstituted glycol group polyethylene glycol groupranging rangingininsize size from fromabout about1 1totoabout about1212ethylene ethyleneglycol glycol units, between units, between 1 1and andabout about 10 ethylene 10 ethylene glycol glycol units, units, about about 2 about 2 about 6 ethylene 6 ethylene glycol glycol units, units, betweenabout between about2 2and and5 5ethylene ethyleneglycol glycolunits, units, between betweenabout about2 2and and4 4ethylene ethyleneglycol glycolunits. units.

[0105] In In

[0105] another another the the embodiment, embodiment, present present disclosureisisdirected disclosure to aa compound directed to which compoundwhich comprises comprises aaPTM PTM group, group, which which binds binds to a to a target target protein protein or polypeptide, or polypeptide, which which is ubiquitinated is ubiquitinated

by an by an ubiquitin ubiquitin ligase ligase and andisis chemically chemicallylinked linkeddirectly directlytotothe theULM ULM group group (such(such as or as CLM) CLM) or through aa linker through linker moiety L, or moiety L, or PTM PTM isisalternatively alternatively aa ULM' group ULM' group (such (such as as CLM') CLM') which which is also is also a a ubiquitin ligase ubiquitin ligase binding moiety, which binding moiety, whichmaymay be the be the samesame or different or different than than the group the ULM ULMasgroup as

103 described above andisislinked aboveand linkeddirectly directly to to the the ULM ULM group or or directly through the the linker moiety; 2023248067 10 Oct 2023 described group directly through linker moiety; and and LL is is aa linker linker moiety moiety as as described above which described above which may maybe be present absentand presentororabsent andwhich which chemically (covalently) links chemically (covalently) links ULM ULM to to PTM, PTM, or a or a pharmaceutically pharmaceutically acceptable acceptable salt, salt, enantiomer, enantiomer, stereoisomer, solvate stereoisomer, solvate or or polymorph thereof. polymorph thereof.

[0106] In In

[0106] certain thethe embodiments, certainembodiments, linker L isa group groupL is linkergroup a group comprising oneone comprising or more or more

covalently connected covalently connected structural structural unitsunits independently independently selectedselected from the from group the group of: consisting consisting of:

* *** ** X N N

*I O R1 R¹ R¹ R1 O * ** N O * *' N * N * R R¹ 0 The XXisis selected The selected from fromthe thegroup groupconsisting consistingofofO,0,N,N,S,S,S(O) S(O)andand SO;SO ; ninteger n 2is is integer fromfrom 1 to1 5; to 5;

N Rl is hydrogen RL¹ is hydrogenor or alkyl, * alkyl, is a mono- is mono-or orbicyclic bicyclicaryl arylor or heteroaryl heteroaryl optionally optionally

substituted substituted with 1-3 substituents with 1-3 substituents selected selectedfrom fromalkyl, alkyl,halogen, halogen,haloalkyl, haloalkyl,hydroxy, hydroxy, alkoxy alkoxy or or

N cyano; cyano; * * is aa mono- is mono-ororbicyclic bicycliccycloalkyl cycloalkyl orora heterocycloalkyl a heterocycloalkyloptionally optionally substituted substituted with 1-3 1-3 substituents substituents selected selected from fromalkyl, alkyl,halogen, halogen,haloalkyl, haloalkyl,hydroxy, hydroxy, alkoxy alkoxy or or

cyano; andthe cyano; and thephenyl phenylring ringfragment fragment can can be optionally be optionally substituted substituted withwith 1, 2 1, or 23 or 3 substituents substituents

selected selected from the grou from the grou consisting consisting of of alkyl, alkyl, halogen, halogen, haloalkyl, haloalkyl,hydroxy, hydroxy, alkoxy and cyano. alkoxy and cyano. InInanan embodiment, thelinker embodiment, the linkergroup group L comprises L comprises up toup 10 to 10 covalently covalently connected connected structural structural units, units, as as described above. described above.

[0107] Although

[0107] the the Although groupgroup ULMULM andgroup and PTM group PTM may be may covalently linked linked be covalently the linker to the tolinker

group throughanyany group through group group which which is appropriate is appropriate and stable and stable to thetochemistry the chemistry of the in of the linker, linker, in preferred aspects preferred aspects of of the the present dislcosure, dislcosure, the the linker linkerisisindependently independently covalently covalently bonded to the bonded to the ULM group ULM group andand thethe PTMPTM group group preferably preferably through through an amide, an amide, ester, ester, thioester, keto thioester, keto group, group, carbamate (urethane), carbon carbamate (urethane), carbonororether, ether,each eachofofwhich which groups groups may may be inserted be inserted anywhere anywhere on theon the

ULM groupand ULM group andPTM PTM group group to provide to provide maximum maximum binding binding of the of the ULMULM groupgroup on ubiquitin on the the ubiquitin ligase andthe ligase and thePTM PTM groupgroup on theon the target target proteinprotein to be degraded. to be degraded. (It that (It is noted is noted that in in certain certain aspects aspects

104

Oct 2023 where the where the PTM PTM group group is aisULM a ULM group,group, the target the target protein protein for degradation for degradation may bemay the be the ubiquitin ubiquitin

ligase itself). In ligase itself). certainpreferred In certain preferredaspects, aspects, the the linker linker may may be be to linked linked to an optionally an optionally substitutedsubstituted

alkyl, alkylene,alkene alkyl, alkylene, alkene or oralkyne group, an alkyne group, an aryl arylgroup or aaheterocyclic group or group on heterocyclicgroup on the theULM and/or ULM and/or

groups. PTMgroups. PTM 2023248067 10

[0108] In additional

[0108] In additional embodiments, embodiments, q isinteger q is an an integer fromfrom 1to 100, 1 to 100, 1 to 1 90, to 90, 1 to1 80, to 80, 1 to70,70,1 1toto 1 to

60, 60, 11 to to 50, 50, 11 to to 40, 40,1 1toto30, 30,1 1toto20, 20,oror1 Ito to10.10.

[0109]

[0109] In certain In certain embodiments, embodiments, the linker the linker (L) (L) is is selected selected from from the theconsisting group group consisting of: of:

2

o 0

OH OH 0

O O0 0 O O

0

oo 10

0 0

o ZI H N

H ZI 0 0 0 H N N O N

0 0-. ~ ~ 0 o 0/ O 0

0 ~ 0 0

105

Oct 2023 0 0 0

2023248067 10

O 0 0 O 0 0 new

N o; ; ;n NVV nvvv

00 0O

ml N o ~~~~ ^^^^

you 0

0 0 ;and ; and

N N, N

[0110] InInadditional

[0110] embodiments,thethe additional embodiments, linker group linkergroup is optionally is optionally substituted substituted (poly)ethyleneglycol havingbetween (poly)ethyleneglycol having between1 and 1 and about about 100100 ethylene ethylene glycol glycol units,between units, between about about 1 and 1 and

about 50 ethylene about 50 ethylene glycol glycolunits, units, between between1 1and andabout about 25 25 ethylene ethylene glycol glycol units, units, between between about about 1 1 and 10 ethylene and 10 ethylene glycol glycolunits, units, between between1 1and andabout about8 ethylene 8 ethylene glycol glycol unitsandand units 1 and 1 and 6 ethylene 6 ethylene

glycol units, units, between between2 and 2 and 4 ethylene 4 ethylene glycol glycol units,or units,or optionally optionally substituted substituted alkyl groups alkyl groups

interdispersed interdispersed with with optionally substituted, substituted, O, 0, N, N, S, S, P P or Si Si atoms. In In certain certain embodiments, embodiments,thethe

linker is substituted linker is withanan substituted with aryl,phenyl, aryl, phenyl, benzyl, benzyl, alkyl, alkyl, alkylene, alkylene, or heterocycle or heterocycle group. group. In certainIn certain

embodiments, thelinker embodiments, the linkermay maybebeasymmetric asymmetric or or symmetrical. symmetrical.

[0111] In any

[0111] In of anythe of embodiments the embodiments of the of the compounds compounds described described herein, herein, the thegroup linker linkermay group may be any be any suitable suitable moiety moietyasasdescribed describedherein. herein.In In oneone embodiment, embodiment, the linker the linker is a is a substituted substituted or or unsubstituted polyethylene glycol group polyethylene glycol groupranging rangingininsize size from fromabout about1 1toto about about1212ethylene ethyleneglycol glycol

106

units, between units, between 1 1and andabout about 10 ethylene 10 ethylene glycol glycol units, units, about about 2 about 2 about 6 ethylene 6 ethylene glycol glycol units, units, betweenabout between about2 2and and5 5ethylene ethyleneglycol glycolunits, units, between betweenabout about2 2and and4 4ethylene ethyleneglycol glycolunits. units. thethe Although

[0112] Although

[0112] (or (or CLMCLM ULM) groupgroup ULM) and PTM PTM group and group may may be be covalently covalently linked linked to the to the

linker linker group through any group through anygroup groupwhich whichis isappropriate appropriateandand stabletotothe stable thechemistry chemistryofofthe thelinker, linker, in in preferred aspects preferred aspects of of the the present disclosure, disclosure, the the linker linkerisisindependently independently covalently covalently bonded to the bonded to the CLM groupandand CLM group thethe PTMPTM group group preferably preferably through through an amide, an amide, ester, ester, thioester, keto thioester, keto group, group, 2023248067

carbamate (urethane), carbon carbamate (urethane), carbonororether, ether, each eachofofwhich which groups groups may may be inserted be inserted anywhere anywhere on theon the

CLM groupand CLM group andPTM PTM group group to to providemaximum provide maximum binding binding of the of the CLMCLM group group on ubiquitin on the the ubiquitin ligase andthe ligase and thePTM PTM groupgroup on theon the target target proteinprotein to be degraded. to be degraded. (It that (It is noted is noted that in in certain certain aspects aspects

where the PTM where the PTM group group is aisULM a ULM group,group, the target the target protein protein for degradation for degradation may bemay the be the ubiquitin ubiquitin

ligase itself). In ligase itself). certainpreferred In certain preferredaspects, aspects, the the linker linker may may be be to linked linked to an optionally an optionally substitutedsubstituted

alkyl, alkyl, alkylene, alkylene,alkene alkene or oralkyne alkyne group, group, an an aryl arylgroup group or or aaheterocyclic heterocyclicgroup group on on the theCLM and/or CLM and/or

PTM groups. PTM groups.

[0113] In certain

[0113] In certain embodiments, embodiments, "L"becan "L" can be linear linear chainschains with linear with linear atoms atoms from 4 from 4 to to 24, the24, the carbon atomininthe carbon atom thelinear linearchain chaincancan be be substituted substituted with with oxygen, oxygen, nitrogen, nitrogen, amide, amide, fluorinated fluorinated

carbon, etc., such carbon, etc., suchasasthethefollowing: following:

N IZ O O O O O,,,--''ONOOO IZ N H N H O H H H ZI H ZI H H N IZ O N N O o O N N0 N O O N H H O

IZ IZ o N H N H H H 0 0 O o N IZ N N O N IZ N Os o IZ N O N IZ N O o H H H H H H H H

o IZ o N H N H H H H ZI H H ZI H N IZ O N NO IZ O N N O N N H H O o

107

2023248067 10 2023

ZI IZ O O H N H N H H O O 0' Oct ZI O IZ o ZI IZ O N HH N H N H N H H H H -/- IZ IZ N HH O N O H H

-/- H ZI H ZI N IZ o N N O HH H H O -/- ZI i'N N N IZ N INIZ N N' IZ N HH HH H H H H

-/-

ZI O IZ IZ O IZ N HH N HH N N H H H H ,or ,or

H H -/-

IZ O ZI IZ N N N H N H o HH H H F FFEFF FF F FEF F F H H IZ IZ ZI 'N N N N N O HH F F F F H H H H

101141

[0114] In certain certainembodiments, "L"can embodiments, "L" canbebenonlinear nonlinearchains, chains,and andcan canbebealiphatic aliphatic or or aromatic aromatic

or or heteroaromatic cyclic heteroaromatic cyclic moieties, moieties, someexamples some examples of "L" of"L" includeinclude but not but not be limited be limited to the to the

following: following:

-1 -- %

O-X-Y-|- HN HN \/ O-X-Y- O-X-Yi HN H O-X-Y-:

F

HN\/HN\/ N HN O-X-Y -O-X-Y-H o-x-y-|- HN F F

HN X Y HN O-X-Y- N N N Y-/-

HN O-X-Y-i N ,N HN -NN N

108

F

N ZI N / X ' \1-N -NFF - X N X H H N H /0H0 N H H FF

IZ N X IZ X ZI X N N H H H -NIZ NN H IZ N N H HHH X H N N N

O OY N NN N -;N \ IZ N NN 1_ N N W HN H N Y N -:-NH H / N _N\-X Y

O IZ N N N N Y.

Y NH .~-H H'o- o | 0

N N NH NH X X-Y --- X -- NH N/\ --- NH NH

wherein: wherein:

'X"inin above X" abovestructures structures can can be be linear linear chain chain with with atoms ranging from atoms ranging from 22 to to 14, 14, and and the the mentioned chaincan mentioned chain cancontain containheteroatoms heteroatomssuch such as as oxygen; oxygen; andand

"Y" in above "Y" in abovestructures structures can can be be O, 0, N, N, S(O) S(O),(n=0, (n=O,1,1,2). 2).

[0115] Exemplary

[0115] PTMs Exemplary PTMs

[0116]

[0116] In aspects of preferred aspects In preferred of the the present disclosure,the presentdisclosure, thePTM PTM group group, which is aa group, group is binds which binds

to target to targetproteins. proteins.Targets Targetsofofthe PTM the PTM group are numerous group are numerous ininkind kindand andare areselected selectedfrom fromproteins proteins that are expressed that are expressedin in a cell a cell such such that that at least at least a portion a portion of theof the sequences sequences is the is found in found cellinand the cell and maybind may bindtotoaa PTM PTM group. group. TheThe term term "protein" "protein" includes includes oligopeptides oligopeptides and and polypeptide polypeptide sequences sequences

of of sufficient sufficient length lengththat thatthey theycan canbind bind to toaaPTM groupaccording PTM group accordingtotothethepresent presentdisclosure. disclosure.Any Any protein in protein in aa eukaryotic systemorora amicrobial eukaryotic system microbialsystem, system,including including a virus,bacteria a virus, bacteriaor orfungus, fungus,as as

109 otherwise herein, are described herein, are targets targets for ubiquitination mediated for ubiquitination by the the compounds compounds according 2023248067 10 2023 otherwise described mediated by according to the present disclosure. Preferably, the target protein is a eukaryotic protein. In certain aspects, to the present disclosure. Preferably, the target protein is a eukaryotic protein. In certain aspects,

Oct the protein the protein binding moiety isis aa haloalkane binding moiety haloalkane (preferably (preferably aa C-C Ci-Cio alkyl alkyl group group whichwhich is substituted is substituted

with at with at least least one halo group, one halo group, preferably preferably a ahalo halogroup groupatatthethedistal distal end endofofthe thealkyl alkylgroup, group,i.e., i.e., away fromthe away from thelinker linker oror CLM CLM group), group), which which may may covalently covalently bind bind to a to a dehalogenase dehalogenase enzymeenzyme in a in a patient or subject or in a diagnostic assay. patient or subject or in a diagnostic assay.

[0117] PTM groups

[0117] PTM groups according according to the present to the present disclosure disclosure include, include, for example, for example, include include any any moiety which binds to a protein specifically (binds to a target protein) and includes the following moiety which binds to a protein specifically (binds to a target protein) and includes the following

non-limiting examples non-limiting examplesof of small small molecule molecule targettarget protein protein moieties: moieties: Hsp90 inhibitors, Hsp90 inhibitors, kinase kinase inhibitors, inhibitors,androgen androgenreceptor receptorinhibitors, HDM2 inhibitors, HDM2 & MDM2 & MDM2 inhibitors, compounds inhibitors, compounds targeting targeting

Human BETBET Human Bromodomain-containing Bromodomain-containing proteins, proteins, HDACHDAC inhibitors, inhibitors, human human lysine lysine methyltransferase inhibitors, methyltransferase inhibitors, angiogenesis angiogenesisinhibitors, inhibitors,nuclear nuclearhormone hormone receptor receptor compounds, compounds,

immunosuppressive compounds, immunosuppressive compounds, and compounds and compounds targeting targeting thehydrocarbon the aryl aryl hydrocarbon receptor receptor (AHR), (AHR),

among numerous among numerous others. others. TheThe compositions compositions described described belowbelow exemplify exemplify some ofsome of the members the members of of these nine these nine types types of of small small molecule moleculetarget targetprotein proteinbinding bindingmoieties. moieties.Such Such small small molecule molecule target target

protein binding protein binding moieties moietiesalso alsoinclude includepharmaceutically pharmaceutically acceptable acceptable salts, salts, enantiomers, enantiomers, solvates solvates

and polymorphs and polymorphsofofthese thesecompositions, compositions,asaswell wellasasother othersmall smallmolecules moleculesthat thatmay may targeta aprotein target protein of interest. of interest. These binding moieties These binding moieties are are linked linked toto the the ubiquitin ubiquitin ligase ligase binding moietypreferably binding moiety preferably through a linker in order to present a target protein (to which the protein target moiety is bound) through a linker in order to present a target protein (to which the protein target moiety is bound)

in in proximity proximity toto theubiquitin the ubiquitin ligase ligase for for ubiquitination ubiquitination and degradation. and degradation.

whichwhich Any protein,

[0118] Any protein,

[0118] bind can to can bind to a protein a protein target target moiety moiety or group or PTM group PTM and andonacted acted or on or degraded degraded by by a ubiquitin a ubiquitin ligase ligase is a is a target target protein protein according according to the disclosure. to the present present disclosure. In general,In general,

target target proteins proteins may include, for may include, forexample, example,structural structuralproteins, proteins,receptors, receptors, enzymes, enzymes,cell cellsurface surface proteins, proteins proteins, proteins pertinent pertinent to to the the integrated integrated function of aa cell, function of cell, including including proteins proteins involved in involved in

catalytic catalytic activity, activity, aromatase activity, motor aromatase activity, motoractivity, activity, helicase helicaseactivity, activity, metabolic metabolicprocesses processes (anabolism andcatabolism), (anabolism and catabolism),antioxidant antioxidantactivity, activity, proteolysis, proteolysis, biosynthesis, biosynthesis, proteins proteins with kinase with kinase

activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase

activity, ligase activity, activity, ligase activity, enzyme enzyme regulator regulator activity, activity, signal signal transducer transducer activity, activity, structural structural moleculemolecule

activity, binding activity (protein, lipid carbohydrate), receptor activity, cell motility, membrane activity, binding activity (protein, lipid carbohydrate), receptor activity, cell motility, membrane

fusion, cell communication, fusion, cell communication, regulation regulation of biological of biological processes, processes, development, development, cell differentiation, cell differentiation,

response to stimulus, behavioral proteins, cell adhesion proteins, proteins involved in cell death, response to stimulus, behavioral proteins, cell adhesion proteins, proteins involved in cell death,

110 involvedinintransport proteins involved transport(including (includingprotein protein transporter activity,nuclear nuclear ion ion transport, 2023248067 10 Oct 2023 proteins transporter activity, transport, transporter activity,channel transporter activity, channel transporter transporter activity, activity, carriercarrier activity, activity, permease permease activity, secretion activity, secretion activity, electron activity, electron transporter transporter activity, activity,pathogenesis, pathogenesis, chaperone regulator activity, chaperone regulator activity, nucleic nucleic acid acid binding activity, binding activity, transcription transcription regulator regulatoractivity, activity, extracellular extracellular organization organizationandand biogenesis biogenesis activity, translation activity, translationregulator regulatoractivity. activity.Proteins Proteins of interest of interest can include can include proteins proteins from eurkaryotes from eurkaryotes and prokaryotes and prokaryotes including including humans humansasastargets targets for for drug drugtherapy, therapy, other other animals, animals, including including domesticated animals, domesticated animals,microbials microbials for for the the determination determination of targets of targets for antibiotics for antibiotics and and other other antimicrobials and antimicrobials and plants, plants, and and even viruses, among even viruses, numerous among numerous others. others.

[0119] In still

[0119] In still other other embodiments, embodiments, the group the PTM PTM isgroup is a haloalkyl a haloalkyl group, wherein group, wherein said said alkyl alkyl group generally ranges group generally rangesininsize size from fromabout about 1 or 1 or 2 carbons 2 carbons to about to about 12 carbons 12 carbons in length, in length, oftenoften

about about 22 to to 10 10 carbons carbonsininlength, length, often often about about 33 carbons carbonstotoabout about8 8carbons carbonsininlength, length,more more often often

about about 44 carbons carbonstotoabout about6 6carbons carbons in in length.TheThe length. haloalkyl haloalkyl groups groups are generally are generally linear linear alkylalkyl

groups (although branched-chain groups (although branched-chainalkyl alkylgroups groupsmay may also also be be used) used) andand areare end-capped end-capped withwith at least at least

one halogengroup, one halogen group,preferably preferablya asingle singlehalogen halogengroup, group,often oftena single a singlechloride chloridegroup. group.Haloalkyl Haloalkyl PT, groups for PT, groups for use use in in the the present present disclosure disclosure are are preferably preferably represented represented by the chemical by the structure chemical structure

-(CH2)v-Halo -(CH)-Halo where where V is v is integer any any integer from from 2 to about 2 to about 12, often 12, often about about 3 to 8, 3 to about about more8,often more often about about 44 to to about about 6. 6. Halo maybebeany Halo may anyhalogen, halogen,but butisispreferably preferably Cl Cl oror Br, Br, more moreoften often Cl. Cl. In another

[0120] In another

[0120] embodiment, embodiment, the present the present disclosure disclosure provides provides a library a library of compounds. of compounds. The The library library comprises morethan comprises more thanoneone compound compound wherein wherein each composition each composition has a of has a formula formula A-B, of A-B, wherein A Ais isa aubiquitin wherein ubiquitinpathway pathway protein protein binding binding moiety moiety (preferably, (preferably, an E3 an E3 ubiquitin ubiquitin ligase ligase moiety as moiety as otherwise otherwisedisclosed disclosedherein) herein) and andB Bisisaa protein protein binding bindingmember memberof of a molecular a molecular library, library,

wherein A Ais iscoupled wherein coupled (preferably, (preferably, through through a linker a linker moiety) moiety) to B,toand B,wherein and wherein the ubiquitin the ubiquitin

pathwayprotein pathway proteinbinding bindingmoiety moiety recognizes recognizes an ubiquitin an ubiquitin pathway pathway protein, protein, in particular, in particular, an E3an E3 ubiquitin ligase, ubiquitin ligase, such as cereblon. such as cereblon. In In aa particular particular embodiment, embodiment,thethe librarycontains library contains a specific a specific

cereblon E3ubiquitin cereblon E3 ubiquitinligase ligase binding bindingmoiety moiety bound bound to random to random targettarget protein protein binding binding elements elements

(e.g., (e.g.,a achemical chemical compound library). As compound library). As such, such, the the target target protein protein is isnot notdetermined determined in inadvance advance and and

the method the canbebeused method can used to to determine determine the the activity activity of of a putativeprotein a putative protein binding binding element element and and its its pharmacologicalvalue pharmacological valueasasa atarget target upon degradationbybyubiquitin upon degradation ubiquitinligase. ligase.

[0121] TheThe

[0121] present present disclosuremaymay disclosure be used be used to treat to treat a number a number of disease of disease states states and/or and/or

conditions, including conditions, including any any disease disease state state and/or and/or condition condition in in which whichproteins proteinsare aredysregulated dysregulatedandand where where aa patient patient would benefit from would benefit fromthe the degradation degradationofofproteins. proteins.

111

[0122] In anInadditional

[0122] an additional aspect, aspect, the the description description provides provides therapeutic therapeutic compositions compositions comprising comprising

an effective amount an effective amount ofofa acompound compound as described as described herein herein or form or salt salt form thereof, thereof, and aand a

pharmaceutically pharmaceutically acceptable acceptable carrier, carrier, additive additive or excipient, or excipient, and optionally and optionally an additional an additional bioactive bioactive agent. Thetherapeutic agent. The therapeuticcompositions compositions modulate modulate protein protein degradation degradation in a patient in a patient or subject, or subject, for for

example, anananimal example, animalsuch suchasasa ahuman, human,andand can can be used be used for for treating treating or or ameliorating ameliorating disease disease states states

or conditions conditions which whichare aremodulated modulated through through the the degraded degraded protein. protein. In certain In certain embodiments, embodiments, the the therapeutic compositions therapeutic compositionsas asdescribed described herein herein may may be toused be used to effectuate effectuate the degradation the degradation of of proteinsofofinterest proteins interestforforthethe treatment treatment or amelioration or amelioration of a disease, of a disease, e.g.,(such e.g., cancer cancer (such as as prostate prostate cancer) and andKennedy's Kennedy's Disease. Disease. In certain In certain additional additional embodiments, embodiments, the is the disease disease is prostate prostate

cancer. cancer.

In alternative

[0123] In alternative

[0123] thethe aspects, aspects, present present disclosure disclosure relatestotoa amethod relates methodforfor treatinga adisease treating disease state or state or ameliorating the symptoms ameliorating the symptoms of of a disease a disease or condition or condition in ainsubject a subject in need in need thereof thereof by by degrading degrading a aprotein proteinororpolypeptide polypeptide through through which which a disease a disease state state or condition or condition is modulated is modulated

comprising administeringtotosaid comprising administering saidpatient patientororsubject subjectananeffective effectiveamount, amount, e.g.,a atherapeutically e.g., therapeutically effective effective amount, of at amount, of at least least one one compound compound asasdescribed describedhereinabove, hereinabove, optionally optionally in in combination combination

with aa pharmaceutically with pharmaceuticallyacceptable acceptablecarrier, carrier,additive additiveororexcipient, excipient,and andoptionally optionallyan an additional additional

bioactive agent, bioactive agent, wherein the composition wherein the compositionisiseffective effective for for treating treating or or ameliorating ameliorating the the disease disease or or disorder or symptom disorder thereofininthe symptom thereof thesubject. subject. The Themethod method according according to to thethe present present disclosuremaymay disclosure

be used be usedtototreat treata alarge largenumber number of disease of disease states states or conditions or conditions includingincluding cancer, by cancer, virtue ofby thevirtue of the administration of effective administration of effective amounts of at amounts of at least leastone onecompound describedherein. compound described herein. The The diseasestate disease state or condition may maybebea adisease diseasecaused caused by by a microbial a microbial agent agent or other or other exogenous exogenous agentagent such such as a as a

virus, virus, bacteria, bacteria,fungus, fungus,protozoa protozoa or or other other microbe or may microbe or bea adisease may be diseasestate, state, which whichisis caused causedbyby overexpression ofprotein, overexpression of a a protein, which which leads leads to a disease to a disease state and/or state and/or condition. condition.

In another

[0124] In another

[0124] aspect, aspect, the description the description provides provides methods methods for identifying for identifying the effects the effects of of the the degradation ofofproteins degradation proteins ofofinterest interest inin aabiological biological system systemusing using compounds compounds according according to the to the presentdisclosure. present disclosure.

[0125] term "target The "target

[0125] The term protein" protein" is used used is to to describe describe a protein a protein or polypeptide, which iswhich or polypeptide, a is a target for target for binding binding totoa acompound compound according according to the to the present present disclosure disclosure and degradation and degradation by by ubiquitin ligase ligase hereunder. hereunder. Such Suchsmall smallmolecule molecule target target protein protein binding binding moieties moieties also also include include

pharmaceutically acceptable pharmaceutically acceptablesalts, salts, enantiomers, enantiomers, solvates solvates and andpolymorphs polymorphsof of these these compositions, compositions,

112 as well asasother othersmall small molecules thattarget may target a protein of interest. Thesemoieties bindingaremoieties are 2023248067 10 Oct 2023 as well molecules that may a protein of interest. These binding linked linked to to CLM CLM ororULM ULM groups groups through through linker linker groups groups L. L.

[0126] Target

[0126] Target proteins proteins whichwhich may may be be bound bound to the to the protein protein targettarget moietymoiety and degraded and degraded by the by the ligase ligase to to which theubiquitin which the ubiquitinligase ligasebinding bindingmoiety moiety is is bound bound include include any protein any protein or peptide, or peptide,

including fragments fragmentsthereof, thereof,analogues analogues thereof, thereof, and/or and/or homologues homologues thereof. thereof. Target Target proteinsproteins

include proteins proteins and andpeptides peptideshaving having anyany biological biological function function or activity or activity including including structural, structural,

regulatory, hormonal, regulatory, hormonal, enzymatic, enzymatic,genetic, genetic,immunological, immunological, contractile, contractile, storage, storage, transportation, transportation,

and signal transduction. and signal transduction. In In certain certain embodiments, embodiments,thethetarget targetproteins proteinsinclude includestructural structural proteins, proteins, receptors, enzymes, receptors, enzymes,cellcell surface surface proteins, proteins, proteins proteins pertinent pertinent to the integrated to the integrated function offunction a cell, of a cell, including proteins proteins involved involvedinincatalytic catalyticactivity, activity, aromatase aromataseactivity, activity,motor motor activity,helicase activity, helicase activity, metabolic activity, processes(anabolism metabolic processes (anabolism and and catabolism), catabolism), antioxidant antioxidant activity, activity, proteolysis, proteolysis,

biosynthesis, proteins biosynthesis, proteins with with kinase kinase activity, activity, oxidoreductase oxidoreductase activity, activity, transferase transferase activity, activity,

hydrolaseactivity, hydrolase activity,lyase lyase activity, activity, isomerase isomerase activity, activity, ligase ligase activity, activity, enzyme activity, enzyme regulator regulator activity, signal transducer signal transduceractivity, activity,structural structuralmolecule molecule activity, activity, binding binding activity activity (protein, (protein, lipid lipid carbohydrate), receptor carbohydrate), receptor activity, activity, cell cellmotility, motility,membrane fusion, cell membrane fusion, cell communication, regulation communication, regulation

of biological biological processes, processes, development, development, cellcell differentiation,response differentiation, response to stimulus, to stimulus, behavioral behavioral

proteins, cell proteins, cell adhesion proteins, proteins adhesion proteins, proteins involved in cell involved in cell death, death, proteins involved inin transport proteins involved transport (including protein protein transporter transporteractivity, activity, nuclear nucleartransport, transport,ionion transporter transporter activity, activity, channel channel

transporteractivity, transporter activity,carrier carrier activity, activity, permease permease activity, activity, secretion secretion activity, activity, electron transporter electron transporter

activity, pathogenesis,chaperone activity, pathogenesis, chaperone regulator regulator activity, activity, nucleic nucleic acidactivity, acid binding bindingtranscription activity, transcription regulatoractivity, regulator activity,extracellular extracellularorganization organization and biogenesis and biogenesis activity, activity, translation translation regulator regulator activity.activity.

Proteins of Proteins of interest interest can can include proteins from eukaryotes proteins from eukaryotesand andprokaryotes, prokaryotes,including including microbes, microbes,

viruses, viruses, fungi fungi and parasites, including humans, and parasites, humans,microbes, microbes, viruses,fungi viruses, fungi andand parasites, parasites, among among

numerousothers, numerous others,asastargets targetsfor fordrug drug therapy,other therapy, other animals, animals, including including domesticated domesticated animals, animals,

microbialsforforthethedetermination microbials determination of targets of targets for antibiotics for antibiotics and antimicrobials and other other antimicrobials andandplants, and plants, and even viruses, even viruses, among numerous among numerous others. others.

[0127] More More

[0127] specifically, specifically, a number a number of drug drug targets of targets for human for human therapeutics therapeutics represent proteinprotein represent targets totowhich targets which protein protein target targetmoiety moiety may be bound may be boundand andincorporated incorporatedinto intocompounds compounds according according

to the to the present present disclosure. disclosure. These Theseinclude includeproteins proteins which which may may be be toused used to restore restore function function in in numerous polygenic numerous polygenic diseases, diseases, including forfor including example B7.1B7.1 example andand B7, B7, TINFRm, TNFR2, TINFRlm, TNFR2,NADPH NADPH

oxidase, oxidase, BclIBax andother BcllBax and otherpartners partners in in the the apotosis apotosis pathway, C5areceptor, pathway, C5a receptor, HMG-CoA HMG-CoA reductase, reductase,

113

PDE PDE V V phosphodiesterase type, type, PDE PDEIV IV phosphodiesterasetype 2023248067 10 Oct 2023

phosphodiesterase phosphodiesterase type4,4,PDE PDE I, I, PDEII, PDEII, PDEIII, PDEIII,

squalene cyclaseinhibitor, squalene cyclase inhibitor, CXCR1, CXCR1, CXCR2, CXCR2, nitric nitric oxide oxide (NO) synthase, (NO) synthase, cyclo-oxygenase cyclo-oxygenase 1, 1, cyclo-oxygenase cyclo-oxygenase 2,2,5HT 5HT receptors,dopamine receptors, dopamine receptors, receptors, G Proteins, G Proteins, i.e.,Gq, i.e., Gq,histamine histaminereceptors, receptors, 5-lipoxygenase, tryptase serine 5-lipoxygenase, tryptase serine protease, protease, thymidylate synthase, purine thymidylate synthase, nucleoside phosphorylase, purine nucleoside phosphorylase, GAPDH trypanosomal, GAPDH trypanosomal, glycogen glycogen phosphorylase, phosphorylase, Carbonic Carbonic anhydrase, anhydrase, chemokine chemokine receptors, receptors,

JAWSTAT, JAW STAT, RXRRXR and similar, and similar, HIV HIV 1 protease, 1 protease, HIV HIV 1 integrase, 1 integrase, influenza,neuramimidase, influenza, neuramimidase, hepatitis B hepatitis reverse transcriptase, B reverse transcriptase, sodium sodiumchannel, channel, multi multi drugdrug resistance resistance (MDR), (MDR), proteinprotein P- P glycoprotein (and glycoprotein (and MRP), MRP), tyrosine tyrosine kinases, kinases, CD23, CD23, CD124, CD124, tyrosine tyrosine kinasekinase p56CD4, p56 lck, lck, CD5, CD4, CD5, IL-2 IL-2 receptor, receptor,IL-I IL-1receptor, TNF-alphaR, receptor, TNF-alphaR,ICAM1, ICAM1, Cat+ channels, channels, VCAM, VLA-4 VCAM, VLA-4 integrin, integrin,

selectins, selectins,CD40/CD40L, newokinins CD40/CD40L, newokinins and and receptors, receptors, inosine inosine monophosphate monophosphate dehydrogenase, dehydrogenase, p38 p38 MAP Kinase,RaslRaflMEWERK MAP Kinase, RaslRaflMEWERK pathway, pathway, interleukin-1 interleukin-1 converting converting enzyme, enzyme, caspase, caspase, HCV,HCV,

NS3 protease, NS3 protease, HCV HCVNS3 NS3 RNA helicase, RNA helicase, glycinamide glycinamide ribonucleotide ribonucleotide formylformyl transferase, transferase,

rhinovirus 3C rhinovirus 3Cprotease, protease,herpes herpes simplex simplex virus-i virus-1 (HSV-I), (HSV-I), protease, protease, cytomegalovirus cytomegalovirus (CMV) (CMV) protease, poly protease, (ADP-ribose)polymerase, poly (ADP-ribose) polymerase, cyclin cyclin dependent dependent kinases, kinases, vascular vascular endothelial endothelial growth growth

factor, oxytocinreceptor, factor, oxytocin receptor, microsomal microsomal transfertransfer protein protein inhibitor,inhibitor, bile acid bile acid inhibitor, transport transport 5 inhibitor, 5 alpha reductase reductase inhibitors, inhibitors, angiotensin angiotensin 11, 11,glycine glycinereceptor, receptor,noradrenaline noradrenaline reuptake reuptake receptor, receptor,

endothelin receptors, endothelin receptors, neuropeptide neuropeptide YYand andreceptor, receptor,estrogen estrogenreceptors, receptors, androgen androgenreceptors receptors(AR), (AR), adenosine receptors, adenosine adenosine receptors, adenosinekinase kinaseand andAMP AMP deaminase, deaminase, purinergic purinergic receptors receptors (P2Y1,(P2Yl, P2Y2, P2Y2,

P2Y4, P2Y6, P2Y4, P2Y6, P2X1-7), P2X1-7), famesyltransferases, farnesyltransferases, geranylgeranyl geranylgeranyl transferase, transferase, TrkA aTrkA a receptor receptor for for NGF,beta-amyloid, NGF, beta-amyloid,tyrosine tyrosinekinase kinase Flk-IIKDR, Flk-IIKDR, vitronectin vitronectin receptor, receptor, integrin integrin receptor, receptor, Her-21 Her-21

neu, telomerase neu, telomeraseinhibition, inhibition,cytosolic cytosolicphospholipaseA2 phospholipaseA2 and and EGF EGF tyrosine receptor receptor kinase. tyrosine kinase. Additional protein Additional protein targets targets include, include, for forexample, example, ecdysone 20-monooxygenase, ecdysone 20-monooxygenase, ion ion channel channel of the of the

GABA gated GABA gated chloride chloride channel, channel, acetylcholinesterase, acetylcholinesterase, voltage-sensitive voltage-sensitive sodium sodium channel channel protein, protein,

calcium release channel, calcium release channel, and andchloride chloridechannels. channels.Still Still further further target target proteins proteins include include Acetyl-CoA Acetyl-CoA

carboxylase, adenylosuccinate synthetase, carboxylase, adenylosuccinate synthetase, protoporphyrinogen protoporphyrinogen oxidase, oxidase, and and enolpyruvylshikimate-phosphatesynthase. enolpyruvylshikimate-phosphate synthase.

Haloalkane

[0128] Haloalkane

[0128] dehalogenase dehalogenase enzymes enzymes are another target target are another of specific of specific compounds compounds

according toto the according thepresent presentdisclosure. disclosure.Compounds Compounds according according to the present to the present disclosure disclosure which which contain chloroalkane chloroalkane peptide peptide binding bindingmoieties moieties(C-C 1 2 often often (C1-C about about C2-C1 oalkyl C-C alkyl halo groups) halo groups) may may be used be used toto inhibit inhibit and/or and/or degrade degradehaloalkane haloalkanedehalogenase dehalogenase enzymes enzymes which which areinused are used in fusion fusion proteins or proteins or related related dioagnostic dioagnostic proteins proteins as as described in PCT/US2012/063401 described in PCT/US2012/063401 filedfiled December December 6, 6,

114

2011 and publishedasasWOWO andpublished 2012/078559 on 14, 14, 2012, June2012, the contents of which is incorporated 2023248067 10 Oct 2023

2011 2012/078559 on June the contents of which is incorporated

by reference by herein. referenceherein.

[0129] TheseThese

[0129] various various protein protein targets may may targets be used be used in screens that that in screens identify identify compound compound moieties moieties

which bindtotothe which bind the protein protein and incorporationof of andbybyincorporation themoiety the moiety into into compounds compounds according to theto the according presentdisclosure, present disclosure,thethelevel level of of activity activity of the of the protein protein may may be be altered altered for therapeutic for therapeutic end end result. result.

The term

[0130] The term

[0130] "protein "protein target target moiety" or PTMoris moiety" usedisto PTM used describe small molecule a smalla molecule to describe which which binds to binds to aa target target protein protein ororother otherprotein proteinororpolypeptide polypeptide of interest of interest andand places/presents places/presents thatthat

proteinororpolypeptide protein polypeptide in proximity in proximity to an to an ubiquitin ubiquitin ligase ligase such thatsuch that degradation degradation of the of the protein or protein or polypeptide bybyubiquitin polypeptide ubiquitinligase ligasemaymay occur. occur. Non-limiting Non-limiting examples examples ofmolecule of small small molecule target target protein binding protein moieties include binding moieties include Hsp90 Hsp90inhibitors, inhibitors, kinase kinase inhibitors, inhibitors, MDM2 inhibitors, MDM2 inhibitors,

compounds targeting Human compounds targeting BETBromodomain-containing Human BET Bromodomain-containing proteins, HDAC proteins, HDAC inhibitors, human inhibitors, human lysine lysine methyltransferase inhibitors, angiogenesis methyltransferase inhibitors, angiogenesis inhibitors, inhibitors,immunosuppressive compounds, immunosuppressive compounds, andand

compounds targeting compounds targeting the the aryl aryl hydrocarbon hydrocarbon receptor receptor (AHR), (AHR),among among numerous numerous others. others. The The

compositions describedbelow compositions described below exemplify exemplify some some of theof the members members of these of these nine nine types types of small of small

moleculetarget molecule target protein. protein.

[0131] Exemplary

[0131] Exemplary protein protein target target moieties moieties according according to the to the present present disclosureinclude, disclosure include, haloalkane halogenase haloalkane halogenase inhibitors, inhibitors, Hsp90 Hsp90 inhibitors, inhibitors, kinasekinase inhibitors, inhibitors, MDM2 inhibitors, MDM2 inhibitors,

compounds targeting compounds targeting Human BETBromodomain-containing Human BET Bromodomain-containing proteins, HDAC proteins, HDAC inhibitors, human inhibitors, human lysine lysine methyltransferase inhibitors, angiogenesis methyltransferase inhibitors, angiogenesis inhibitors, inhibitors,immunosuppressive compounds, immunosuppressive compounds, andand

compounds targetingthe compounds targeting thearyl arylhydrocarbon hydrocarbon receptor(AHR). receptor (AHR).

[0132] The compositions

[0132] The compositions described described below exemplify below exemplify some of the of the of somemembers members of these these types of types of small molecule moleculetarget targetprotein proteinbinding binding moieties. moieties. SuchSuch smallsmall molecule molecule target target proteinprotein bindingbinding

moieties also moieties also include pharmaceutically acceptable salts, pharmaceutically acceptable salts, enantiomers, solvates and polymorphs enantiomers, solvates polymorphs ofof

these compositions, these compositions,asaswell well as as other other small small molecules molecules that target that may may target a protein a protein of interest. of interest.

Referenceswhich References which are cited are cited hereinbelow hereinbelow are incorporated are incorporated by herein by reference reference herein in their in their entirety. entirety.

[0133] I. I.

[0133] Heat Heat Shock Shock Protein 90 90 Protein (HSP90) (HSP90) Inhibitors: Inhibitors:

[0134] HSP90HSP90

[0134] inhibitors inhibitors as used as used herein herein include, include, but are are not but not limited limited to: to:

[0135] 1. The

[0135] The HSP90 1. HSP90 inhibitors inhibitors identified identified in Vallee, in Vallee, et al., et al., "Tricyclic "Tricyclic Series Series of Heat of Heat Shock Shock

Protein 90 Protein 90 (HSP90) (HSP90)Inhibitors InhibitorsPart PartI: I: Discovery DiscoveryofofTricyclic TricyclicImidazo[4,5-C]Pyridines Imidazo[4,5-C]Pyridines as Potent as Potent

Inhibitors ofofthethe Inhibitors HSP90 HSP90Molecular MolecularChaperone Chaperone (2011) (2011) J.Med.Chem. J.Med.Chem. 54: 54: 7206, 7206, including includingYKB YKB

(N-[4-(3H-imidazo[4,5-C]Pyridin-2-yl)-9H-Fluoren-9-yl]-succinamide): (N-[4-(3H-imidazo[4,5-C]Pyridin-2-yl)-9H-Fluoren-9-yl]-succinamide).

115

Oct 2023 0 O HN HN NH - /\ O O 2023248067 10

N N NH NH

[01361

[0136] N N derivatized where derivatized where aa linker linkergroup groupLL or or aa -(L-CLM) -(L-CLM)

group is attached, group is attached, for forexample, example, via via the theterminal terminalamide amide group; group;

[0137] 2. The

[0137] The HSP90 2. HSP90 inhibitor inhibitor p54 (modified) p54 (modified) (8-[(2,4-dimethylphenyl)sulfanyl]-3]pent-4 (8-[(2,4-dimethylphenyl)sulfanyl]-3]pent-4-

yn-1-yl-3H-purin-6-amine): yn-1-yl-3H-purin-6-amine):

NH NH2 N N N N S S N N N N

[01381

[0138] derivatized where derivatized where aa linker linker group group LL or or aa -(L-CLM) -(L-CLM) group group isisattached, attached,forforexample, example, via terminal via the the terminal acetylene acetylene group; group;

[0139] 3. The

[0139] The HSP90 3. HSP90 inhibitors inhibitors (modified) (modified) identified identified in Brough, in Brough, et al., et al., "4,5-Diarylisoxazole "4,5-Diarylisoxazole

HSP90Chaperone HSP90 Chaperone Inhibitors:Potential Inhibitors: PotentialTherapeutic TherapeuticAgents Agents forfor thethe Treatment Treatment of Cancer", of Cancer",

J.MED.CHEM. J.MED.CHEM. vol: vol: 51, 51, pag:pag:196 (2008), 196 (2008), including including the the compound compound 2GJ (5-[2,4-dihydroxy-5-(1 2GJ (5-[2,4-dihydroxy-5-(1-

methylethyl)phenyl]-n-ethyl-4-[4-(morpholin-4-ylmethyl)phenyl]isoxazole-3-carboxamide) methylethyl)phenyl]-n-ethyl-4-[4-(morpholin-4-ylmethyl)phenyljisoxazole-3-carboxamide)

having the structure: having the structure:

116

Oct 2023

C_0 N

NN 2023248067 10

0

N HO Ho | H H O-NN

[01401

[0140] OH derivatized, where derivatized, where aa linker linker group group LL or or aa -(L-CLM) -(L-CLM)

group isisattached, group attached,forforexample, example, via amide via the the amide group group (at the (at theoramine amine at the or at the alkyl alkyl group group on the on the

amine); amine);

[0141] 4. The

[0141] 4. HSP90 The HSP90 inhibitors inhibitors (modified) (modified) identified identified in Wright, in Wright, et al., et al., Structure-Activity Structure-Activity

Relationships in Relationships in Purine-Based Inhibitor Binding Purine-Based Inhibitor Bindingtoto HSP90 HSP90 Isoforms, Isoforms, Chem Chem Biol. Biol. 20042004

Jun;11(6):775-85, including Jun;11(6):775-85, including the the HSP90 HSP90 inhibitorPU3 inhibitor PU3 having having thethe structure: structure:

NH NH2 NA N N N NN N/ N 0 O -0 0

[0142]

[0142] derivatized derivatized where where aa linker linker group L or group L or -(L-CLM) -(L-CLM) is is

attached, forexample, attached, for example,via via the the butyl butyl group; group; and and

[0143]

[0143] 5. 5. The HSP90inhibitor The HSP90 inhibitorgeldanamycin geldanamycin ((4E,6Z,8S,9S,1OE,12S,13R,14S,16R)-13 (4E,6Z,8S,9S,10E,12S,13R,14S,16R)-13-

hydroxy-8,14,19-trimethoxy-4,10,12,16-tetramethyl-3,20,22-trioxo-2-azabicyclo[16.3.1] hydroxy-8,14,19-trimethoxy-4,10,12,16-tetramethyl-3,20,22-trioxo-2-azabicyclo[163.1]

(derivatized) (derivatized) or or any any of of its itsderivatives (e.g. derivatives 17-alkylamino-17-desmethoxygeldanamycin (e.g. ("17 17-alkylamino-17-desmethoxygeldanamycin ("17-

AAG")oror 17-(2-dimethylaminoethyl)amino-17-desmethoxygeldanamyci AAG") 17-(2-dimethylaminoethyl)amino-17-desmethoxygeldanamycin ("17-DMAG")) ("17-DMAG")) (derivatized, (derivatized, where where a a linker linker group group L L or or a-(L-CLM) group a-(L-CLM) group is isattached, attached,for forexample, example,via viathe the amide amide group). group).

[0144]

[0144] I. II. Kinase andPhosphatase Kinase and Phosphatase Inhibitors: Inhibitors:

Kinase

[0145] Kinase

[0145] inhibitors inhibitors as used as used herein herein include, but but include, notnot are are limited to:to: limited

[0146]

[0146] 1. Erlotinib 1. Derivative Tyrosine Erlotinib Derivative Tyrosine Kinase Inhibitor: Kinase Inhibitor:

117

HN HN O R '0 -Zz N TO NN

where where RRis is aa linker linker group or aa -(L-CLM) L or group L group -(L-CLM) group attached, example,viaviathetheether attached,forforexample, ethergroup; group; 2023248067

[0147]

[0147] 2. The kinase inhibitor sunitinib (derivatized): 2. The kinase inhibitor sunitinib (derivatized):

R R F F /N N IZ

H 0H ZI O N

[01481

[0148] H derivatized where derivatized where RRis is aa linker linker group group L or aa -(L-CLM) L or -(L-CLM)

group attached, group attached, forfor example, example, to the to the pyrrole pyrrole moiety; moiety;

[0149]

[0149] 3. Kinase Inhibitor sorafenib (derivatized): 3. Kinase Inhibitor sorafenib (derivatized):

0 O CI O R O IZ "aoKNNH 0 R CF N N IZ O N CF N N

[0150]

[0150] H H derivatized where R is a linker derivatized where R is a linker

group group LL or or aa -(L-CLM) group -(L-CLM) group attached, attached, forfor example, example, to to thethe amide amide moiety; moiety;

[0151]

[0151] 4. The kinase inhibitor desatinib (derivatized): 4. The kinase inhibitor desatinib (derivatized):

CI CI

NH N 0 O S NH NH N N N N R derivatized R derivatized where where RRis is aa linker linker group Lora-(L-CLM) group Lor a-(L-CLM) attached, forexample, attached, for example, to the to the pyrimidine; pyrimidine;

[0152] 5. The

[0152] 5. kinase The kinase inhibitor inhibitor lapatinib lapatinib (derivatized): (derivatized):

118

Oct 2023 F F

CI CI

O O S 2023248067 10

HN HN HN HN 0 NO N O

[01531

[0153] N derivatized where derivatized where aa linker linker group L or group L or a-(L- a-(L CLM) is isattached, group CLM) group attached,for for example, example,via viathe the terminal methylofofthe terminal methyl the sulfonyl sulfonyl methyl methylgroup; group;

[0154] 6. The

[0154] 6. kinase The kinase inhibitor inhibitor U09-CX-5279 U09-CX-5279 (derivatized): (derivatized):

H ZI H N N N N N N HO O Ho N NH N NH 0C O CF derivatized where a linker group L or a -(L-CLM)

[0155]

[0155] CF3 derivatized where a linker group L or a -(L-CLM) group is attached, group is attached, for forexample, example,viavia thethe amine amine (aniline), (aniline), carboxylic carboxylic acid acid or amine or amine alpha to alpha to

cyclopropyl group, oror cyclopropyl cyclopropyl group, cyclopropylgroup; group;

[0156] The kinase 7. kinase

[0156] 7. The inhibitors inhibitors identified identified in Millan, in Millan, et al., et al., Design and and Design Synthesis Synthesis of Inhaled of Inhaled

P38 Inhibitors for P38 Inhibitors forthe theTreatment Treatmentofof Chronic ChronicObstructive ObstructivePulmonary PulmonaryDisease, Disease,J.MED.CHEM. J.MED.CHEM.

vol:54, vol:54, pag:7797 (2011), including pag:7797 (2011), including the the kinase kinase inhibitors inhibitors YlW and Y1W and YlX Y1X (Derivatized) (Derivatized) having having the the

structures: structures:

0 o N N N IZ IZ N~ H H H H S

N N N-N N-N

119

YIX(1-ethyl-3-(2-{[3-(1-methylethyl)[1,2,4]triazolo[4,3-a]pyridine-6

[0157] YIX(1-ethyl-3-(2-{[3-(1-methylethyl)[1,2,4]triazolo[4,3-a]pyridine-6-

[0157]

yl]sulfanyl}benzyl)urea, derivatized yl]sulfanyl}benzyl)urea, derivatized where where aa linker linker group group LLor or a-(L-CLM) a-(L-CLM) group group is attached, is attached, forfor

example, via the example, via the propyl ¹propyl group; group;

oO N-N N-N 2023248067

HN HN N IZ N H H NN ~ s~NN N N NN CS- S

YIW YIW 1-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-{[3-(1-methylethyl)[1,2,4]triazolo[4,3-a]pyridin-6-yl]sulfanyl}benzyl)urea 1-(3-tert-buty-1-pheny-1H-pyrazol-5-y)-3-(2-{[3-(1-methylethyl)[1,2,4triazolol4,3-a]pyridin-6-yl]sultany)berzyi)urea

derivatized derivatized where where aa linker linker group group LL or or aa -(L-CLM) group -(L-CLM) group is is attached,forforexample, attached, example, preferablyviavia preferably

either the i-propyl either the i-propylgroup groupor or thethe t-butyl t-butyl group; group;

[0158] 8. The

[0158] The kinase 8. kinase inhibitors inhibitors identified identified in Schenkel, et al., et in Schenkel, Discovery of Potentofand al., Discovery Potent and Highly SelectiveThienopyridine Highly Selective Thienopyridine Janus Janus Kinase Kinase 2 Inhibitors 2 Inhibitors J.Chem., J. Med. Med. 2011, Chem., 54 2011, (24), 54 (24),

pp 8440-8450, pp 8440-8450,including includingthe thecompounds compounds6TP 6TP and (Derivatized) and OTP OTP (Derivatized) havinghaving the structures: the structures:

HN HN O O 0=6=0 S-NH

S S1 - O N N S0

NH2 NH 6TP 6TP 4-amino-2-[4-(tert-butylsulfamoyl)phenyl]-N-methylthieno[3,2-c]pyridine-7-carboxamide 4-amino-2-[4-(tert-butylsulfamoyl)phenyl-N-methylthienol[3,2-c]pyridine-7-carboxamide Thienopyridine1919 Thienopyridine

derivatized wherea alinker derivatized where linkergroup group L aor-(L-CLM) L or a -(L-CLM) group group is is attached, attached, for example, for example, via the via the

terminal methyl terminal groupbound methyl group boundtotoamide amide moiety; moiety;

H1N HN O O S N1 N O N N H2 NH 120

Oct 2023 OTP OTP 4-amino-N-methyl-2-[4-(morpholin-4-yl)phenyl]thieno[3,2-c]pyridine-7-carboxamide 4-amino-N-methyl-2-[4-(morpholin-4-yl)phenylhieno[3,2-c]pyridine-7-carboxamide Thienopyridine 88 Thienopyridine

derivatized wherea alinker derivatized where linkergroup group L orL aor a -(L-CLM)group -(L-CLM)group is attached, is attached, for example, for example, via the via the terminal methyl terminal groupbound methyl group boundtotothe theamide amidemoiety; moiety; 2023248067 10

9. kinase

[0159] 9. The

[0159] The kinase inhibitors inhibitors identified identified in Van Van et in Eis, et al., Eis,al., "2,6-Naphthyridines "2,6-Naphthyridines as potent as potent

and selective inhibitors and selective inhibitors of of the the novel novel protein protein kinase kinase C isozymes",Biorg. C isozymes", Biorg.Med. Med. Chem. Chem. Lett.2011 Lett.2011

Dec 15;21(24):7367-72,including Dec 15;21(24):7367-72, includingthe thekinase kinaseinhibitor inhibitor07U 07Uhaving having thestructure: the structure: NH NH2 HN HN N N N Nzz - N N 07U 07U 2-methyl-N-1--[3-(pyridin-4-yl)-2,6-naphthyridin-1-yl]propane-1,2-diamine 2-methyl-N-1~-[3-(pyridin-4-yl)-2,6-naphthyridin-1-yilpopane-1,2-diamine

derivatized wherea alinker derivatized where linkergroup group L orL aor a -(L-CLM)group -(L-CLM)group is attached, is attached, for example, for example, via the via the secondary amineororterminal secondary amine terminalamino aminogroup; group;

[0160] The kinase 10. kinase

[0160] 10. The inhibitors inhibitors identified identified in Lountos, in Lountos, et al., et al., "Structural Characterizationofof "StructuralCharacterization Inhibitor Inhibitor Complexes with Complexes with Checkpoint Checkpoint Kinase Kinase 2 (Chk2), 2 (Chk2), a Drug aTarget Drug for Target forTherapy", Cancer Cancer Therapy", J.STRUCT.BIOL.vol:176, J.STRUCT.BIOL. vol:176,pag:292 pag:292(2011), (2011),including including the the kinase kinase inhibitor inhibitor YCF havingthe YCF having the structure: structure:

H H HH H ZI H N N N / N - NN N N HO' Ho N N 0 N' N N 'OH O OH NH 2 NH N A N& N NH NH N N

[01611

[0161] H H derivatized where derivatized where a linker linker group group L Lorora -(L-CLM) a -(L-CLM) group group is attached, is attached, for example, for example, via of via either either of the terminal the terminal

hydroxyl groups; hydroxyl groups; 11. kinase

[0162] 11. The

[0162] The kinase inhibitors inhibitors identified identified in Lountos, in Lountos, et al., et al., "Structural Characterizationofof "StructuralCharacterization Inhibitor Complexes Inhibitor with Complexes with Checkpoint Checkpoint Kinase Kinase 2 (Chk2), 2 (Chk2), a Drug aTarget Drug for Target forTherapy", Cancer Cancer Therapy", J.STRUCT.BIOL. vol:176, J.STRUCT.BIOL. vol:176,pag:292 pag:292(2011), (2011),including includingthe thekinase kinase inhibitors inhibitors XK9 XK9and andNXPNXP (derivatized) having (derivatized) having thethe structures: structures:

121

HN OH 2023248067 10 Oct 2023

HN OH Y-NH NH N0 2 HN-NH NO ~NH ZI

HN/ N-NH N HN

O XK9 XK9 N-{4-[(1E)-N-(N-hydroxycarbamimidoyl)ethanehydrazonoyl]phenyl}-7-nitro-1H-indole-2-carboxamide; N-{4-[(1E)-N-(N-hydroxycarbamimidoyl)ethanehydrazonoyl]phenyl)-7-nitto-1H-indole-2-carboxamide

H ZI H N N

NH NH 0 O

N NH NH HN NH2 NXP NH NXP

[0163]

[0163] N-{4-[(1E)-N-CARBAMIMIDOYLETHANEHYDRAZONOYLIPHENYL}1H-INDOLE-3-CARBOXAMIDE N-{4-[(1E)-N-CARBAMIMIDOYLETHANEHYDRAZONOYL]PHENYL}-1H-INDOLE-3-CARBOXAMIDE derivatized wherea alinker derivatized where linkergroup group L aor-(L-CLM) L or a -(L-CLM) group group is is attached, attached, for example, for example, via the via the

terminal terminal hydroxyl group(XK9) hydroxyl group (XK9)or or thehydrazone the hydrazone group group (NXP); (NXP);

[0164]

[0164] 12. The The 12. kinasekinase inhibitor inhibitor afatinib afatinib (derivatized) (derivatized) (N-[4-[(3-chloro-4 (N-[4-[(3-chloro-4- fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)-2 fluorophenyl)amino]-7-I[(3S)-tetrahydro-3-furanylloxy]-6-quinazolinyl]-4(dimethylamino)-2-

butenamide)(Derivatized butenamide) (Derivatizedwhere where a linkergroup a linker groupL L or or a a -(L-CLM) -(L-CLM) group group is attached, is attached, for for example, example,

via the aliphatic via the aliphaticamine amine group); group);

[0165] 13.13.

[0165] The kinase The kinase inhibitor inhibitor fostamatinib fostamatinib (derivatized) (derivatized) ([6-({5-fluoro-2-[(3,4,5 ([6-({5-fluoro-2-[(3,4,5-

trimethoxyphenyl)amino]pyrimidin-4-yl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H trimethoxyphenyl)amino]pyrimidin-4-yl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-

pyrido[3,2-b]-1,4-oxazin-4-yl]methyl pyrido[3,2-b]-1,4-oxazin-4-yl]methyl disodium disodium phosphate phosphate hexahydrate) hexahydrate) (Derivatized (Derivatized where awhere a linker linker group group L or aa -(L-CLM) L or group -(L-CLM) group is isattached, attached,for for example, example,via viaa amethoxy methoxy group); group);

[0166] 14. kinase

[0166] 14. The The inhibitor kinase inhibitor gefitinib gefitinib (derivatized) (derivatized) (N-(3-chloro-4-fluoro-phenyl)-7 (N-(3-chloro-4-fluoro-phenyl)-7-

methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine): methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine):

122

F F HN HN CI CI R R N N

[01671

[0167] 0 O N derivatized where derivatized where aa linker linkergroup groupLL or or aa -(L-CLM) -(L-CLM)

group is attached, group is attached, for forexample, example, via via aamethoxy or ether methoxy or ether group; group; 2023248067

[01681

[0168] 15. 15. The kinase The kinase inhibitor inhibitor lenvatinib lenvatinib (derivatized) (derivatized) (4-[3-chloro-4 (4-[3-chloro-4-

(cyclopropylcarbamoylamino)phenoxy]-7-methoxy-quinoline-6-carboxamide) (cyclopropylcarbamoylamino)phenoxy]-7-methoxy-quinoline-6-carboxamide) (derivatized (derivatized

where where a alinker linkergroup groupL or L aor-(L-CLM) a -(L-CLM) group group is is attached, attached, for example, for example, via the cyclopropyl via the cyclopropyl

group); group);

[0169] 16. kinase

[0169] 16. The The kinase inhibitor inhibitor vandetanib vandetanib (derivatized) (derivatized) (N-(4-bromo-2-fluorophenyl)-6 (N-(4-bromo-2-fluorophenyl)-6-

methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine) (derivatized methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine) (derivatized where where a linker a linker

group group LL or or aa -(L-CLM) group -(L-CLM) group is is attached,for attached, forexample, example,viaviathethemethoxy methoxyor or hydroxyl hydroxyl group); group);

[0170] 17. The

[0170] 17. kinase The kinase inhibitor inhibitor vemurafenib vemurafenib (derivatized) (derivatized) (propane-1-sulfonic (propane-1-sulfonic acid acid {3-[5- {3-[5 (4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluoro-phenyl}-amide), (4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluoro-phenyl}-amide),

derivatized wherea alinker derivatized where linkergroup groupL or L aor-(L-CLM) a -(L-CLM) group group is is attached, attached, for example, for example, via the via the

sulfonyl sulfonyl propyl group; propyl group;

[0171] 18. kinase

[0171] 18. The The kinase inhibitor inhibitor Gleevec Gleevec (derivatized): (derivatized):

N H N N HN N N HN HN

[0172]

[0172] O O R derivatized where derivatized where RRas asaalinker linker group group LLorora-(L-CLM) a-(L-CLM) group group isisattached, attached,forforexample, example, via amide via the the amide group group or or via via the the amine aniline aniline amine group; group;

[0173] 19. kinase

[0173] 19. The The kinase inhibitor inhibitor pazopanib pazopanib (derivatized) (derivatized) (VEGFR3 (VEGFR3 inhibitor): inhibitor):

123

R R 'NH NH N N NN N N N NN

[01741

[0174] where RRisisa alinker derivatized where derivatized groupL Lorora a- linker group (L-CLM) group (L-CLM) group attached,forforexample, attached, example, to to phenylmoiety thephenyl the moiety viavia or or theaniline the anilineamine group; aminegroup;

[0175]

[0175] Thekinase 20. The 20. inhibitor AT-9283 kinaseinhibitor Aurora (Derivatized)Aurora AT-9283(Derivatized) Kinase Kinase Inhibitor Inhibitor

0 O H N IZ N HN HN H H R R N SN N-NH N NH IZ N

[01761

[0176] H H N where RR is where is aa linker linkergroup groupL Lorora a-(L-CLM) group -(L-CLM) group

attached, forexample, attached, for example, to the to the phenyl phenyl moiety); moiety);

[0177]

[0177] 21. Thekinase 21. The TAE684 inhibitor TAE684 kinaseinhibitor (derivatized) ALK (derivatized)ALK inhibitor inhibitor

CI CI N 00 H N NH O O HN N NH S O

[01781

[0178] R where where RRisis aa linker linker group groupLLorora a-(L-CLM) -(L-CLM) group group

attached, forexample, attached, for example, to the to the phenyl phenyl moiety); moiety);

[0179]

[0179] 22. The kinase inhibitor nilotanib (derivatized) Abl inhibitor: 22. The kinase inhibitor nilotanib (derivatized) Abl inhibitor:

HN HN N N N N O N N NH NH O N N 0 R R

[01801

[0180] F3C FC derivatized where derivatized where RRis is aa linker linker group group LL

or or aa -(L-CLM) group -(L-CLM) group attached,forforexample, attached, example,totothe thephenyl phenylmoiety moiety or or theaniline the anilineamine aminegroup; group;

124

[0181] 23.23.

[0181] Kinase Kinase Inhibitor NVP-BSK805 InhibitorNVP-BSK805 (derivatized) JAK2 (derivatized)JAK2 Inhibitor Inhibitor

0 O N N F F F F

N N 2023248067

NN N-R ''

[0182]

[0182] N derivatized where derivatized where R Risisa alinker linkergroup group L aor-(L- L or a -(L CLM) group CLM) group example,to tothe attached,forforexample, attached, thephenyl phenylmoiety moiety or or thethediazole diazolegroup; group;

[0183]

[0183] 24. Kinase 24. Kinase Inhibitor Inhibitor crizotinib crizotinib Derivatized Derivatized Alk Inhibitor Alk Inhibitor

R R N N N N N NH 0 NH2 O CI CI CI CI

[0184]

[0184] F F derivatized where derivatized where RRis is aa linker linker group L or group L or aa -(L-CLM) -(L-CLM) group group

attached, forexample, attached, for example, to the to the phenyl phenyl moiety moiety or the or the diazole diazole group; group;

25. Kinase

[0185] 25. Kinase

[0185] Inhibitor Inhibitor JNJ FMS FMS (derivatized) JNJ(derivatized) Inhibitor Inhibitor

O HN'O1 O HN O R N 0 N O N IZ N N N N N H H

[01861

[0186] where RRisis aa linker derivatized where derivatized groupL Lorora a- linker group (L-CLM) group (L-CLM) group attached,forforexample, attached, example, to to thephenyl the phenyl moiety; moiety;

The kinase 26. kinase

[0187] 26. The

[0187] inhibitor inhibitor foretinib foretinib (derivatized) MetMet (derivatized) Inhibitor Inhibitor

125

H ZI H H ZI H Ra R N N N N1 0" O 0 0F F O R,0 R

[01881

[0188] O N N derivatized where RRisis aa linker derivatized where linker 2023248067 group group LL or or aa -(L-CLM)group -(L-CLM)group for for attached, attached, example, example, to the to the phenyl phenyl moiety moiety or a or a hydroxyl hydroxyl or ether or ether

group on the group on the quinoline quinoline moiety; moiety;

[0189]

[0189] 27. The 27. Theallosteric allosteric Protein Protein Tyrosine PhosphataseInhibitor Tyrosine Phosphatase Inhibitor PTP1B PTP1B (derivatized): (derivatized):

0 O HN HN R R N 00 N S IZ S S N' N H N IZ S N H |0 V-0s O H

O0 O Br Br

[0190]

[0190] Br Br OH derivatized where a a derivatized where linker linker group group L or aa -(L-CLM) L or group -(L-CLM) group is isattached, attached,for for example, example,atatR, R, as as indicated; indicated;

[0191] 28. The

[0191] 28. inhibitor The inhibitor of SHP-2 of SHP-2 DomainDomain of Tyrosine of Tyrosine Phosphatase Phosphatase (derivatized): (derivatized):

OMe OMe

0 O

R HN R R N / S N \-NN

[0192]

[0192] S derivatized where derivatized where aa linker linkergroup groupL Lorora a-(L-CLM) -(L-CLM)

group group isisattached, attached,forforexample, example, at R; at R;

[0193]

[0193] Theinhibitor 29. The 29. (derivatized) of inhibitor (derivatized) of BRaf BRaf (BRafV 6 0 0 E)/MEK:

126

Oct 2023 R R F. HN S O O CIO CI O 2023248067 10

F F IZ N N N

[01941

[0194] N H H wherea alinker derivatized where derivatized L or groupL or linkergroup a- a (L-CLM) group (L-CLM) group is is forexample, attached,for attached, example,atatR;R; 30. Inhibitor

[0195] 30. Inhibitor

[0195] (derivatized) (derivatized) of Tyrosine of Tyrosine Kinase Kinase ABL ABL Mew. Me

HN HN NH NH R N N N 0N O O N N R N , N

[01961

[0196] derivatized where derivatized wherea alinker linkergroup group L or a-(L-CLM) L or group a-(L-CLM) group is is attached,for attached, example,atatR;R; forexample,

[0197]

[0197] 31. The 31. The kinase inhibitor OSI-027 kinase inhibitor OSI-027 (derivatized) mTORC1/2 (derivatized) mTORC1/2 inhibitor inhibitor

// O

NH NH NH NH2

NN N N N/N N N N /R R ZI

[01981

[0198] 0 O where a alinker derivatized where derivatized linker group groupL Lor ora-(L-CLM) groupgroup a-(L-CLM) is is attached, forexample, attached, for example, at R; at R;

[0199] 32. The

[0199] The kinase 32. kinase inhibitor inhibitor OSI-930 OSI-930 (derivatized) (derivatized) c-Kit/KDR c-Kit/KDR inhibitor inhibitor

127

Oct 2023 OCF 3 OCF

HN\' HN S S

o 2023248067 10

NH NH /N N

0

[02001

[0200] R- R o derivatized where derivatized where aa linker linker group group LLorora-(L-CLM) a-(L-CLM) group attached,forforexample, group isisattached, example, R; and atand at R;

[0201]

[0201] 33. Thekinase 33. The OSI-906(derivatized) inhibitor OSI-906 kinaseinhibitor IGF1R/IR (derivatized) IGF1R/IR inhibitor inhibitor

N N

NH2 NH2

N N N N

[0202]

[0202] R derivatized where derivatized where aa linker linkergroup groupLL or ora-(L-CLM) a-(L-CLM)

group group isisattached, attached,forforexample, example, at R. at R.

[0203] Wherein,

[0203] Wherein, in ofany in any theofembodiments the embodiments described described in sections I-XVII, I-XVII, in sections "R" designates "R" designates a a site site for forattachment attachment of ofaalinker linkergroup groupL Lorora a -(L-CLM)group -(L-CLM)group ononthe thepiperazine piperazinemoiety. moiety.

[0204] III.

[0204] HDM2/MDM2 III. HDM2/MDM2 Inhibitors: Inhibitors: inhibitors inhibitors HDM2/MDM2

[0205] HDM2/MDM2

[0205] as used as used herein herein but include, include, are not are not limited but limited to: to:

[0206] 1. The

[0206] The HDM2/MDM2 1. HDM2/MDM2 inhibitors identified identified inhibitors in in et Vassilev, al., In et Vassilev, vivo In vivo activation al.,activation of of the the p53 pathway by p53 pathway by small-molecule small-molecule antagonists antagonists of of MDM2, SCIENCE MDM2, SCIENCE vol:303, vol:303, pag:844-848 pag:844-848

(2004), and Schneekloth, (2004), and Schneekloth,etetal., al., Targeted Targetedintracellular intracellular protein protein degradation degradationinduced inducedby by a small a small

molecule: En molecule: Enroute routetotochemical chemical proteomics, proteomics, Bioorg. Bioorg. Med.Med. Chem. Chem. Lett. Lett. 18 18 (2008) (2008) 5904-5908, 5904-5908,

128

including (or additionally) including (or additionally) the thecompounds compounds nutlin-3, nutlin-3, nutlin-2, nutlin-2, and nutlin-1 and nutlin-1 (derivatized) (derivatized) as as described below, described below, as well as well as all as all derivatives derivatives and analogs and analogs thereof: thereof:

CI CI

0 O 6 !!!!,

O N N 2023248067

H N HN N " IIII \ CI CI N

(derivatized where a alinker (derivatized where linkergroup groupL or L aor-(L-CLM)group a -(L-CLM)group is attached, is attached, for example, for example, at the at the methoxy groupororasasa ahydroxyl methoxy group hydroxylgroup); group); Br Br

o O N N N N Br HO N N Br HO N N

O O (derivatized where a alinker (derivatized where linkergroup groupL or L or a -(L-CLM) a -(L-CLM) group group is is attached, attached, for example, for example, at the at the methoxy groupororhydroxyl methoxy group hydroxyl group); group);

CI CI O .....

C >NN N N N N "III / C CI

N O 0 O O

(derivatized (derivatized where where a alinker linker group groupL L or or a -(L-CLM) a -(L-CLM) group group is attached, is attached, for example, for example, via the via the

methoxygroup methoxy groupororasasa ahydroxyl hydroxylgroup); group);and and

129

[0207] 2. 2.

[0207] Trans-4-Iodo-4'-Boranyl-Chalcone Trans-4-Iodo-4'-Boranyl-Chalcone

0 O

OOH B OH B OH OH

[0208] (derivatized

[0208] (derivatizedwhere wherea alinker linker group group LL or or aa aa linker linker group group L L or or a-(L-CLM) group is a-(L-CLM) group is 2023248067

attached, attached, for for example, via aa hydroxy example, via group). hydroxy group).

IV.Compounds

[0209] IV.

[0209] Compounds Targeting Human TargetingHuman BETBET Bromodomain-containing Bromodomain-containing proteins: proteins:

In certain

[0210] In certain

[0210] embodiments, "PTM" "PTM" embodiments, can be ligands bindingbinding can be ligands to Bromo- to Bromo- and Extra-terminal and Extra-terminal

(BET) proteins BRD2, (BET) proteins BRD3 BRD2, BRD3 andand BRD4. BRD4. Compounds Compounds targeting targeting HumanHuman BET Bromodomain BET Bromodomain-

containing proteins include, containing proteins include, but are are not not limited limited to to the the compounds associatedwith compounds associated withthe thetargets targetsasas described below, where described below, where"R" "R"oror"linker" "linker"designates designatesa asite site for for linker linker group group L or a-(L-CLM) L or group a-(L-CLM) group

attachment, for for example: example:

[0211] 1. JQl,

[0211] 1. JQ1, et et Filippakopoulos Filippakopoulos al.al.Selective Selective inhibition BET bromodomains. inhibition ofofBET Nature bromodomains. Nature

(2010): (2010):

R R S IZsS Is N N R I"N_ N N N R "" N R / NN NN O-R N N CI CI N O R N O R O X = Cl, Br, F, H, X = CI, Br, F, H,

R S \ N bond,or bond, chemical or aa chemical R S moiety coupling the N N N the S N XN o moiety CLM CLM toto coupling the PTM the PTM S N N O B R O X CI, Br, F, H N N H, aa lower R RR == H, % alkyl, loweralkyl, NN Linker Linker X = CI, Br, F, H R a bond, ororaa chemcial a bond, chemcial moietycoupling moiety couplingthe the \ CLMtoto the CLM the PTM PTM X X N N -N N OO S S 0 CONH 2 \ O O0 CONH O -' , 7- Linker Linker X= X = CI, Br, F, CI, Br, F, H H Linker-N Linker N -N N X= C1, Br, F, H X = CI, Br, F, H NN N N

x X xX

130

Oct 2023

N N -N N S O o O 0 X N o H H Linker-N Linker -N N X = CI, Br, F, H X N N =CI, Br, F, H N H N H N 2023248067 10

x Linker Linker X or or

N S N x_ xi S N N N N 0 X o N N H N H H

Linker Linker

X=H,F X=H,F

[0212]

[0212] 2. Nicodeme I-BET, Nicodeme 2. I-BET, et et Supressionofof al.al.Supression Inflammation by by Inflammation a Synthetic a Synthetic Histone Histone Mimic. Mimic.

Nature(2010). Nature (2010). Chung Chunget etal. al. Discovery Discoveryand and Characterization Characterization of of Small Small Molecule Molecule Inhibitors Inhibitors of the of the

BET Family Bromodomains. BET Family Bromodomains.J.J. Med MedChem. Chem.(2011): (2011): R R O R \ \ R / N N N /

-N N / N N /

JH HNN -NN N N HN-R HN-R CI N O R O R O R R N X == CI, X CI, Br, Br, F, F, H, H, N N N or aa chemical bond, or bond, chemical R / N N 0 moiety coupling moiety the coupling the R O CLMto to the the PTM CLM PTM NH N R R == H, R H,a alower loweralkyl, alkyl, bond, or aa bond, or aa chemcial chemcial moiety coupling the moiety coupling the x CLM to CLM the PTM to the PTM X

[0213] 3. 3.

[0213] Compounds Compounds described described in Hewings in Hewings et al. et al. 3,5-DimethylisoxazolesAct 3,5-Dimethylisoxazoles Actas asAcetyl- Acetyl lysine lysine Bromodomain Ligands. Bromodomain Ligands. J. J. Med. Med. Chem. Chem. (2011) (2011) 54 6761-6770. 54 6761-6770.

Oct 2023 R R HO HO Ho Ho 9O Z-O

- /\ -- N / \ - / -- NOO N N 2023248067 10 0 O OR R

[0214] 4. 4.

[0214] I-BET151, I-BET151, Dawson al. Inhibition et al.et Inhibition Dawson of BET of BET Recruitment Recruitment to Chromatin as anas to Chromatin an Efective Efective Treatment for MLL-fusion Treatment for MLL-fusion Leukemia. Leukemia. Nature Nature (2011): (2011):

R Linker Linker R

N N N N N N

O R IIIIIIIIII O N N N NH NH N N 0 O NH NH O O N N N N N / N N NI N NI N N N O O o

N N IIIIIIIII. Linker Linker

N N ON O N

N ~ N N N 0 O

[0215]

[0215] 5. 5. Carbazole type (US Carbazole type (US 2015/0256700) 2015/0256700) 0 O 0 o NH2 NH2 o -nNH o NH N N -Linker Linker -R N N R N N Linker Linker

102161

[0216]

[0217]

[0217] 6. 6. Pyrrolopyridone type (US Pyrrolopyridone type (US2015/0148342) 2015/0148342)

132

Linke \ Linker, R N / N N N N N Linker .- Linker N F F F O O F O\ \ O

F F / \ N-.. F F / \ N-.. N N N N N H H 0 O H O 2023248067

[02181

[0218]

[0219]

[0219] 7. 7. Tetrahydroquinoline type (WO Tetrahydroquinoline type (WO 2015/074064) 2015/074064)

0 N N N-R Linker Linker

N

[0220]

[0220] O

[0221]

[0221] 8. 8. Triazolopyrazine type (WO Triazolopyrazine type (WO2015/067770) 2015/067770)

N N N N N N N N N N N N N N N N N N R" N N N Linker Linker N N R Linker Linker

[02221

[0222] o 0'

[0223]

[0223] 9. 9. Pyridone type (WO Pyridone type (WO2015/022332) 2015/022332)

N N N N o Linker Linker N

[0224]

[0224]

[0225]

[0225] 10. Quinazolinone 10. type(WO Quinazolinone type (WO 2015/015318) 2015/015318)

R R HN O o70NN N Linker Linker .- l NH NH

[0226]

[0226] 1O O 0

133

[0227] 11.11.

[0227] Dihydropyridopyrazinonetype Dihydropyridopyrazinone type(WO (WO 2015/011084) 2015/011084)

N O N HN HN N N ,-0 O A6 /N N

Linker 2023248067 O NLinker O N

[0228]

[0228] H H

[0229] (Where

[0229] (Where R or LRororlinker, L or linker, in each in each instance, instance, designates designates a site fora attachment, site for attachment, for for example, ofaa linker example, of linker group L or group L or aa -(L-CLM) group). -(L-CLM) group).

[0230] In any

[0230] In any aspect aspect or embodiment or embodiment described described herein, herein, the claimed the claimed structure structure the may the PTM PTMbe may be composed composed ofof tricyclic diazepine tricyclic diazepine or or tricyclic tricyclic azepine azepine as asa aBET/BRD4 targetingmoiety BET/BRD4 targeting moiety (PTM-a), (PTM-a),

where thedashed where the dashed lines lines indicate indicate the linker the linker connection connection trajectory trajectory and and three three sites are sites aretodefined to defined

which linkers may which linkers maybebeattached: attached:

' Z, A Z A Y Y 1 N 1

N0 Y3 Y O B B N N

PTM-a PTM-a

wherein: wherein:

A and A andBBare are independently independentlyananaromatic aromaticring, ring,aaheteroaromatic heteroaromaticring, ring, aa 5-membered 5-membered carbocyclic, carbocyclic, aa 6-membered carbocyclic,a a5-membered 6-membered carbocyclic, 5-membered heterocyclic, heterocyclic, a 6-membered a 6-membered

heterocyclic, heterocyclic, a athiophene, thiophene, a pyrrole, a pyrrole, a pyrazole, a pyrazole, a pyridine, a pyridine, a pyrimidine, a pyrimidine, a pyrazine, a pyrazine,

optionally substituted optionally substituted by by alkyl, alkyl, aloxy, aloxy, halogen, halogen, nitrile nitrile or another or another aromatic aromatic or or heteroaromatic ring, where heteroaromatic ring, where AAis is fused fused to to the the central centralazepine azepine (Yl=C) or diazepine (Y1=C) or diazepine (Y1 (Y1= =

N) moiety; N) moiety; Yl, Y2, Y1, Y2, and andY3 Y3and andY4Y4 cancan be be carbon, carbon, nitrogen nitrogen or or oxygen oxygen for for to to form form a fused a fused 5-membered 5-membered

aromatic ringas astriazole aromatic ring triazoleor or isoxazole; isoxazole; and and

ZI is methyl, Z1 is methyl,ororlower lower alkyl alkyl group. group.

134

The fragment

[0231] The fragment

[0231] of PTM-a of PTM-a as BET/BRD4 moiety is moiety targeting targeting as BET/BRD4 is described described in the literature in the literature

(WO 2016/069578; WO2014/001356; (WO 2016/069578; W02014/001356; W02016/050821; WO2016/050821; WO 2015/195863; WO 2015/195863; WO 2014/128111). WO 2014/128111).

In any

[0232] In any

[0232] aspect aspect or embodiment or embodiment described hereinherein described comprising comprising the structure the structure CLM-L-PTM CLM-L-PTM-

a, a, PTM-a canbeberepresented PTM-a can representedbybythe thefollowing followinggeneral generalstructures, structures, where wheredashed dashedline lineindicates indicates aa possible linker possible linker connection connection point. point. In Instructure structurePTM-aa through PTM-ai, PTM-aa through PTM-ai,the thesubstitution substitution pattern pattern of of X and YYcan X and canbebemono- mono-or or bis-substitution. bis-substitution. 2023248067

S S )

1 N N N -N N NI IN 0 X= F, Br, Cl, F, X = CI, Br, H, H, CN, methyl, acetylene, CN, methyl, acetylene,methoxy methoxy N

X X PTM-aa PTM-aa

S S _ NN O 0 O N S N X= F, Br, Cl, F, X = CI, Br, H, H, CN, methyl, acetylene, CN, methyl, acetylene,methoxy methoxy

PTM-ab PTM-ab x X YN Y N NN N NN Cl, F, X == CI, X F, Br, Br, H, H, CN, CN, methyl, methoxy,acetylene methyl,methoxy, acetylene IN N Y: mono-orordi-substitution, Y: mono- di-substitution, YY= Me,OMe, = Me, OMe, N-methypyrazole/imidazole N-methypyrazole/imidazole

x X PTM-ac PTM-ac

_N N Y O0 O acetylene X = CI, F, Br, H, CN, methyl, methoxy, acetylene X = Cl, F, Br, H, CN, methyl, methoxy, N ! N Y: mono- Y: mono-orordi-substitution, di-substitution, YY == Me, Me,OMe, OMe, N-methylpyrazole/imidazole N-methylpyrazole/imidazole

X PTM-ad PTM-ad

135

-N N N O O O X = CI, F, Br, X = Cl, F, Br, H, H, CN, CN, methyl, methyl, methoxy, acetylene methoxy,acetylene N N NR IZR R = lower alkyl, aryl, substituted aryl R = lower alkyl, aryl, substituted aryl N N H H

x X PTM-ae PTM-ae 2023248067

S N N N N YN 0 N O R= lower alkyl, aryl, substituted aryl N R = lower alkyl, aryl, substituted aryl

. H-R N R IZ O. o H PTM-af PTM-af

N N Y N N N N o Y: mono- or di-substitution, Y = Me, OMe, N-methylpyrazole/imidazole N-methylpyrazole/imidazole \ O Y: mono- or di-substitution, Y = Me, OMe, N ', N'R ZI R N N H R R ==lower alkyl, aryl, substituted aryl lower alkyl, aryl, substituted aryl H

Or O PTM-ag PTM-ag

YNN N N N N N -N O IN N O Cl, F, XX ==CI, Br, H, F, Br, H, CN, methyl, acetylene, CN, methyl, methoxy acetylene,methoxy

X PTM-ah X PTM-ah

N N N N N N N N NO IN N XX ==Cl, CI, F, Br, H, F, Br, H, CN, methyl, acetylene, CN, methyl, methoxy acetylene,methoxy

X X PTM-ai PTM-ai

136

Oct 2023

[0233] In any

[0233] In any aspect aspect or embodiment or embodiment described described herein, herein, the structures the structures of PTM-a as theas of PTM-a the BET/BRD4 targeting BET/BRD4 targeting moiety moiety includes, includes, wherein wherein the the dashed dashed lineline indicates indicates thethe connection connection point point

between the BET/BRD4 between the BET/BRD4 targeting targeting moiety moiety and linkers: and the the linkers:

SS N S S NS S N 2023248067 10

N NIN N / N N I N N N N N N ) N 0N 0N N N O N N O N O N CI CI F F PTM-al PTM-a1 PTM-a2 PTM-a2 PTM-a3 PTM-a3

S S S NN S N N / N NN Z-Z N N -N N N N N O N N 0 N O NN ON o N O N N NNC NC PTM-a4 PTM-a4 PTM-a5 PTM-a5 PTM-a6 PTM-a6

S S S S N S N N N N / NN N, / N N N/ NN N N N N N N 0 0 0 o SN-j NN O N NNs'/ N N O N N N N N O N N ON N N N N PTM-a7 PTM-a7 PTM-a8 PTM-a8 PTM-a9

Z-Z

N 137 N N N N N N N -N -NN -NN N NN o N o N N N N C1 F CI F PTM-a1O PTM-a10 PTM-all PTM-a11 PTM-a12 PTM-a12

\/ Z-Z

\/N NN Z-Z ~ NN Z-z N N N N -N N )-N0N N N -~ ~O 00 O 0 N N N N NC NC PTM-a13 PTM-a13 PTM-al4 PTM-a14 PTM-a15 PTM-a15

N 2023248067

Z-Z

O N N N N N -N N N N -N N N N N Q N N O O ~ N~-O N--~ N N N CI CI CI

PTM-a16 PTM-a16 PTM-al7 PTM-a17 PTM-a18 PTM-a18

-NN -NN -NN 0 00 o O O O o O

-NN N N N N

CI CI F cl PTM-a19 F PTM-a20 cl PTM-a21 PTM-a21 PTM-a19 PTM-a20

-N N N %N % N N 0 00 o O 0 / o 0 0 \ X 0 O O O SN N N N N NC N/

NC NC PTM-a22 PTM-a23 PTM-a23 PTM-a24 PTM-a24 PTM-a22

-NN% -NN N N N N -0

o o O O o o N o N N N N "~N N

CI CI o -0 PTM-a25 cl PTM-a26 cl PTM-a27 PTM-a27 PTM-a25 PTM-a26

138

Oct 2023

-N-N N N N A\ N N\N 'NN \ 0 O -N N N O 0 0 N O N '.N N N N N N 2023248067 10 N O 0"

/ CI CI PTM-a28 CI PTM-a29 PTM-a29 PTM-a30 PTM-a30 PTM-a28

S S N SO N S o Z-Z N -N N 1 N N/ N N/ '\/N N N N 0 -NN N SN -/"I OI r Q N O N O Oz N 0N N IZ N HN- HN H H HN HN PTM-a31 PTM-a31 PTM-a32 PTM-a32 PTM-a33 PTM-a33

-N N NN N -6N- N - C '- -NN O O 0/ 0 O 0 N~ N o o o N N "J N N IZ IZ N\ N N-- IZ N N N H H H H H

CI CI PTM-a34 PTM-a35 PTM-a36 PTM-a36 PTM-a34 PTM-a35

[0234]

[0234] V. HDAC V. HDAC Inhibitors: Inhibitors:

[0235]

[0235] HDAC Inhibitors HDAC Inhibitors (derivatized) (derivatized) include, but arebut include, not are not limited limited to: to:

[0236]

[0236] 1. Finnin, 1. al. Structures M.S.S. etet al. Finnin, M. of Histone Structures of Histone Deacetylase Homologue DeacetylaseHomologue to theto BoundBound the TSA andSAHA TSA and SAHA Inhibitors. Inhibitors. Nature Nature 40, 40, 188-193 188-193 (1999). (1999).

OH O H H R R'N N OH O ZI H N OH HN N N'OH O HN OH O O O R R

(Derivatized where where "R" "R"designates designatesa asite site for for attachment, attachment, for for example, example, ofofaa linker linker group group LL or or aa - (L-CLM) group); and (L-CLM) group); and

139 formula (I) PCT of of WO0222577("DEACETYLASE W00222577 10 Oct 2023

[0237] as as Compounds

[0237] 2.2.Compounds defined by formula defined by (I) PCT ("DEACETYLASE INHIBITORS")(Derivatized INHIBITORS") (Derivatizedwhere wherea alinker linker group group LLoror a a-(L-CLM) -(L-CLM) group group is attached,for is attached, for example, viathe example, via the hydroxyl hydroxylgroup); group);

[0238] VI.VI.

[0238] Human Human Lysine Lysine Methyltransferase Methyltransferase Inhibitors: Inhibitors:

LysineLysine

[0239] HumanHuman

[0239] Methyltransferase Methyltransferase inhibitors inhibitors include, include, but but are not not limited arelimited to: to:

[0240]

[0240] 1. Chang 1. Chang al. al. et et Structural Structural Basis for for Basis G9a-Like G9a-Like protein LysineLysine protein Methyltransferase Methyltransferase

Inhibition Inhibition by by BIX-1294. Nat.Struct. Struct. Biol. Biol. (2009) 16(3) 312. 312. 2023248067

BIX-1294. Nat. (2009) 16(3)

N- N-R N-R N O N N N N J ,0 O N :N-rN

N N O O HN HN H N HN H N N N N'R R

[0241] wherewhere (Derivatized

[0241] (Derivatized "R" designates "R" designates a site a site for for attachment, attachment, for example, for example, of of a linker a linker group group LL or or aa -(L-CLM) group); -(L-CLM) group);

[0242] F. etF.aletDiscovery 2. Liu,

[0242] 2. Liu, al Discovery of a 2,4-Diamino-7-aminoalkoxyquinazoline of a 2,4-Diamino-7-aminoalkoxyquinazoline as as a Potent a Potent and Selective Inhibitor and Selective Inhibitor of ofHistone Histone Methyltransferase G9a. J. Methyltransferase G9a. J. Med. Chem.(2009) Med. Chem. (2009) 52(24) 52(24) 7950. 7950.

N- | N-R N-R N N OO N N NNJN N -NN O O N N N N

/ -N N 0O / N O O O HN HN HN N N HN N N'R R (Derivatized

[0243] (Derivatized

[0243] wherewhere "R" designates "R" designates a potential a potential site for for attachment, siteattachment, for example, for example, of a of a linker linker group group L or a -(L-CLM) L or group); -(L-CLM) group);

3. Azacitidine

[0244] 3. Azacitidine

[0244] (derivatized) (derivatized) (4-amino--p-D-ribofuranosyl-1,3,5-triazin-2(1H)-one) (4-amino-1-ß-D-ribofuranosyl-1,3,5-triazin-2(1H)-on.

(Derivatized where wherea alinker linkergroup groupL or L or a -(L-CLM) a -(L-CLM) group group is attached, is attached, for example, for example, via the via the

hydroxyoror amino hydroxy aminogroups); groups);and and

[0245] 4. Decitabine

[0245] 4. Decitabine (derivatized) (derivatized) (4-amino--(2-deoxy-b-D-erythro-pentofuranosyl)-1, (4-amino-1-(2-deoxy-b-D-erythro-pentofiuranosyl)-1., 3, 3, 5-triazin-2(1H)-one) (Derivatized where 5-triazin-2(1H)-one) (Derivatized wherea alinker linker group groupL Lorora a-(L-CLM) -(L-CLM) groupgroup is attached, is attached, for for

example, viaeither example, via either of of the the hydroxy groupsororat hydroxy groups at the the amino aminogroup). group).

140

Inhibitors: 2023248067 10 Oct 2023

[0246] VII.

[02461 VII. Angiogenesis Angiogenesis Inhibitors:

Angiogenesis

[0247] Angiogenesis

[0247] inhibitors inhibitors include, include, but are not not but are limited to: to: limited

[0248] 1. GA-1

[0248] 1. GA-1 (derivatized) (derivatized) and derivatives and derivatives and analogs and analogs thereof, thereof, having having the the structure(s) structure(s)

and binding to and binding to linkers linkers as as described described in in Sakamoto, et al., Sakamoto, et al., Development Development ofofProtacs Protacsto target cancer to target cancer-

promoting proteins promoting proteins for forubiquitination ubiquitination andand degradation,Mol Mol degradation, Cell Proteomics Cell Proteomics 2003 2003 Dec;2(12):1350-8; Dec;2(12):1350-8;

[0249] 2. Estradiol

[0249] 2. Estradiol (derivatized), (derivatized), which which may may be be bound bound to a linker to a linker group group L or a L or a -(L-CLM) -(L-CLM)

group group asasisisgenerally generallydescribed described in Rodriguez-Gonzalez, in Rodriguez-Gonzalez, et al.,etTargeting al., Targeting steroid steroid hormone hormone

receptors for receptors for ubiquitination ubiquitination and and degradation in breast degradation in breast and prostate cancer, and prostate cancer, Oncogene (2008)27,27, Oncogene (2008)

7201-7211; 7201-7211;

[0250] 3. Estradiol,

[0250] 3. Estradiol, testosterone testosterone (derivatized) (derivatized) and related and related derivatives, derivatives, including but notbut including not limited to to DHT DHTandand derivatives derivatives andand analogs analogs thereof, thereof, having having the structure(s) the structure(s) and and binding binding to a to a

linker linker group group L or aa -(L-CLM) L or group -(L-CLM) group as as generally generally described described in in Sakamoto, Sakamoto, et al., et al., Development Development of of

Protacs to target Protacs to target cancer-promoting cancer-promoting proteins proteins for ubiquitination for ubiquitination and degradation, and degradation, Mol CellMol Cell

Proteomics2003 Proteomics 2003Dec; Dec; 2(12):1350-8; 2(12):1350-8; andand

[0251] 4. Ovalicin,

[0251] 4. Ovalicin, fumagillin fumagillin (derivatized), (derivatized), and and derivatives derivatives and and analogs analogs thereof, thereof, having having the the structure(s) structure(s) and and binding to aa linker binding to linker group group LL or oraa -(L-CLM) -(L-CLM) group group asgenerally as is is generally described described in in

Sakamoto,etetal., Sakamoto, al., Protacs: Protacs: chimeric chimericmolecules molecules thatthat target target proteins proteins to the to the Skpl-Cullin-F Skp1-Cullin-F box box complex forubiquitination complex for ubiquitination and anddegradation degradationProc Proc Natl Natl Acad Acad SciSci USA. USA. 20012001 Jul 17;98(15):8554-9 Jul 17;98(15):8554-9

and United States and United States Patent Patent No. 7,208,157. No. 7,208,157.

VIII.Immunosuppressive

[0252] VIII.

[0252] Compounds: ImmunosuppressiveCompounds: Immunosuppressive

[0253] Immunosuppressive

[0253] compounds compounds include, include, but are not are not to: but limited limited to:

[0254] 1. AP21998

[0254] 1. AP21998 (derivatized), (derivatized), havinghaving the structure(s) the structure(s) and binding and binding to a linker to a linker group group L or L or a -(L-CLM) group -(L-CLM) group as as is is generallydescribed generally described in in Schneekloth, Schneekloth, et et al.,Chemical al., Chemical Genetic Genetic Control Control of of

Protein Levels: Selective Protein Selective in in Vivo TargetedDegradation, Vivo Targeted Degradation,J.JAM. AM.CHEM. CHEM. SOC. SOC. 2004,3748- 2004, 126, 126, 3748 3754; 3754;

[0255] 2. 2.

[0255] Glucocorticoids(e.g., Glucocorticoids (e.g., hydrocortisone, hydrocortisone, prednisone, prednisone, prednisolone, prednisolone, and and methylprednisolone) (Derivatizedwhere methylprednisolone) (Derivatized where a linkergroup a linker groupL or L ora -(L-CLM) a -(L-CLM) groupgroup is to isbound, to bound, e.g. e.g.

to any to of the any of the hydroxyls) and beclometasone hydroxyls) and beclometasonedipropionate dipropionate (Derivatized (Derivatized where where a linker a linker group group or aor a -(L-CLM)is isbound, -(L-CLM) bound, e.g.totoaaproprionate); e.g. proprionate);

141

-(L-CLM) groupororaa -(L-CLM) can can group be 2023248067 10 Oct 2023

[0256] 3. 3.

[0256] Methotrexate Methotrexate (Derivatized where (Derivatizedwhere a linkergroup a linker group be

bound,e.g. bound, e.g.totoeither eitherofofthe theterminal terminal hydroxyls); hydroxyls);

[0257] 4. 4.

[0257] Ciclosporin Ciclosporin (Derivatized where (Derivatizedwhere a linker groupor ora a-(L-CLM) a linkergroup -(L-CLM) group group can can be be bound,e.g. bound, e.g.atatany anyof of the the butyl butyl groups); groups);

[0258] 5. 5.

[0258] Tacrolimus Tacrolimus and and (FK-506) (FK-506) rapamycin rapamycin (Derivatized wherea alinker (Derivatizedwhere or aa - group LL or linker group (L-CLM) group (L-CLM) group cancan be be bound, bound, e.g.e.g. at at oneofof one themethoxy the methoxy groups); groups); andand

[0259] 6. 6.

[0259] Actinomycins Actinomycins (Derivatized (Derivatized where where a linker groupLLoror aa -(L-CLM) a linkergroup canbebe groupcan -(L-CLM)group bound,e.g. bound, e.g.atatone oneof of the the isopropyl isopropyl groups). groups).

[0260] IX.IX.

[0260] Compounds Compounds targeting targeting the aryl the aryl hydrocarbon hydrocarbon receptor receptor (AHR): (AHR):

Compounds

[0261] Compounds

[0261] the the targeting targeting arylaryl hydrocarbon hydrocarbon receptor receptor (AHR) (AHR) include, are are but but include, not not

limited to: limited to:

[0262] 1. Apigenin

[0262] 1. Apigenin (Derivatized (Derivatized in in a away waywhich which bindstoto aa linker binds linker group group LL or ora a-(L-CLM) -(L-CLM)

group as is group as is generally generally illustrated illustrated in in Lee, Lee, et et al., al.,Targeted Targeted Degradation of the Degradation of the Aryl Aryl Hydrocarbon Hydrocarbon Receptor by Receptor by the thePROTAC Approach:A AUseful PROTAC Approach: UsefulChemical ChemicalGenetic Genetic Tool, Tool, ChemBioChem Volume ChemBioChem Volume

8, Issue 8, Issue 17, 17,pages pages 2058-2062, November 2058-2062, November 23,23, 2007); 2007); andand

[0263] 2. 2.

[0263] SR1 SRI and LGCO06 and LGC006 (derivatized (derivatized such such that that a a linker linker group group L orLa or a -(L-CLM) -(L-CLM) is is bound), as bound), as described describedininBoitano, Boitano,etetal., al., Aryl ArylHydrocarbon Hydrocarbon Receptor Receptor Antagonists Antagonists Promote Promote the the ExpansionofofHuman Expansion Human Hematopoietic Hematopoietic Stem Stem Cells,Cells, Science Science 10 September 10 September 2010:329Vol. 2010: Vol. no.329 5997no. 5997 pp. 1345-1348. pp. 1345-1348.

[0264] X. X.

[0264] Compounds Compounds targeting targeting RAF Receptor RAF Receptor (Kinase): (Kinase):

0

F, F HN S HN'S R R O O \ F F IZ N' N N N H H

[0265]

[0265] PLX4032 PLX4032

[0266] (Derivatized

[0266] (Derivatizedwhere where "R""R" designatesa site designates a sitefor for linker linker group LL or or -(L-CLM) -(L-CLM)group group attachment, for for example). example).

whichwhich Any protein,

[0267] Any protein,

[0267] bind can to can bind to a protein a protein target target moiety moiety or group or PTM group PTM and andonacted acted or on or degraded degraded by by an an ubiquitin ubiquitin ligase ligase (e.g., (e.g., RAF) RAF) is a target is a target proteinprotein according according to thedisclosure. to the present present disclosure.

142

Oct 2023

[0268] anyany

[0268] In In or or aspect aspect embodiment embodiment described described herein,the herein, PTMtargets the PTM and/or binds targets and/or RAF binds RAF (i.e., (i.e.,a aRaf Raf or or BRaf targeting moiety). BRaf targeting moiety). For For example, example, in any in any aspect aspect or embodiment or embodiment described described

herein, the herein, the PTM may PTM may comprise comprise a chemical a chemical group group selected selected from from the group the group of chemical of chemical structures structures

consisting consisting of of PTM-Ia orPTM-Ib: PTM-Ia or PTM-Ib: 2023248067 10

RPTM1

XpTM XPTM RPTM2 RPTM2 WPTM WT YPTM RPTM3 M Y ,PTM-RPTM3

ZPTM ZPTM RPTM4 RPTM4

N NN

HO N N Ho PTM-Ia PTM-Ia or or

RPTM1 RPTM1

XpTM XPTM P T RPTM2 RPTM2 WPTM WPTM- YPTM-RPTM3 "YPTM-RPTM3 VPTM ZPTM ZPTM XPTM35 XPTM35 RPTM4 RPTM4

RRPTM5 RRPTM5 //o H HN

S

O XPTM3B6 XPTM36 XPTM38 XPTM38 N XPTM37 XPTM37

PTM-Ib PTM-Ib .

wherein: wherein:

double dotted bonds double dotted bondsare are aromaric aromaricbonds; bonds; VPTM, WPTM, VPTM, WPTM,XPTM, YPTM, XPTM, ZPTM YPTM, ZPTM is ofone the is one of the following following combinations: combinations: C, N, C, CH, CH,N,N,C; N,C, C; N, C, N, N, CH, N, CH,C;C;C,C,O,O,C,C,CH, CH, C; C; C, C, S, S, C, C, CH,CH, C; CH, C; C, C, CH, C, O,C,C; O,C,C;CH,C, C,CH, S, C, C; S, C, C; CH,C,N,CH, N, CH, C; N, CH, C; N, CH, C, CH, CH, C, C; C, CH, C; C, CH, CH, C, C, CH, CH, N; N; N, N, C, CH, C; N, CH, C,C,CH, N,N,C,CH,C;N,CH,C,N,C; N, C;C,C, CH, C, N, N; N, C, N, N; C, N, C, C, CH, CH,N; N;C,C,N,N,C,C,N, N, C;C;and andC,C,N,N,N,N,N,N,C;C; XPTM35, XPTM35, XPTM36, XPTM36, XPTM37, andXPTM38 XPTM37, and areindependently XPTM38are fromCHCH selectedfrom independentlyselected andand N; N;

143

RPTM1 RPTM1 isiscovalently covalently joined totoaa ULM, joined ULM, a achemical chemical linkergroup linker group (L),a CLM, (L), a CLM, an ILM, an ILM, a a VLM, VLM, MLM, MLM, a aULM', ULM',a aCLM', CLM',a aILM', ILM',a aVLM', VLM',a aMLM', MLM',or or combinationthereof; combination thereof; is hydrogen, RPTM2 is RPTM2 hydrogen,halogen, halogen,aryl, methyl, aryl, ethyl, ethyl, methyl, OCH 3 , OCH, NHCH 3 or or NHCH M1-CH 2-CH 2-M2, M1-CH-CH-M2, wherein M1isis CH, wherein M1 CHO2 , and 0 and NH, NH, and and M2 is M2 is hydrogen, hydrogen, alkyl, alkyl, cycliccyclic alkyl,alkyl, aryl aryl or or

heterocycle; heterocycle;

RPTM3 is absent, RPTM3 is absent, hydrogen, hydrogen,aryl, aryl, methyl, methyl,ethyl, ethyl, other other alkyl, alkyl, cyclic cyclic alkyl, alkyl, OCH, OCHNHCH 3 , NHCH or 3 or M1-CH 2 -CH 2-M2, wherein M1 isM1 CH,isO CH 0 and andNH, andhydrogen, M2 is hydrogen, alkyl, cyclic 2023248067

M1-CH-CH-M2, wherein 2 , NH, and M2 is alkyl, cyclic

alkyl, aryl or alkyl, aryl or heterocycle; heterocycle; is hydrogen, RPTM4 is RPTM4 hydrogen,halogen, halogen,aryl, methyl, aryl, ethyl, ethyl, methyl, OCH 3 , OCH, NHCH 3 or or NHCH M1-CH 2-CH 2-M2, M1-CH-CH-M2, wherein M1 wherein M1isis CH, CHO2 , and 0 and NH, NH, and and M2 is M2 is hydrogen, hydrogen, alkyl, alkyl, cycliccyclic alkyl,alkyl, aryl aryl or or heterocycle; and heterocycle; and

RPTM5 isisselected RPTM5 selected from the group from the consisting of group consisting of

F* F FF F* N N F N---- N F F [: NF*F N--- N - --- N---- N

N----- N - ----- 0/ ----- F-- F N

----- F N---- N ----- F F N----- N FF F N

N F ----- F HOF-N N--- N_ F HO N---- N N N Holl!!!!!

HO HO HO HO HO Ho

HO HO

[0269] In In

[0269] anyany ororembodiment aspect aspect embodiment describedherein, described PTM the PTM herein, the may may comprise comprise a chemical a chemical

group selected from group selected from the the group group of ofchemical chemicalstructures structures consisting consisting of of PTM-IIa PTM-IIa ororPTM-IIb: PTM-IIb:

144

RPTM5a RPTM5a RPTM6a RPTM6a RPTM8

O RPTM7 RPTM7 O XPTM3 XPTM3 RPTM9 RPTM9 RPTM6 RPTM6 XPTM2 XPTM2 XPTM4 XPTM4

XPTM1 XPTM XPTM5 PTM5 2023248067

XPTM6 XPTM6 RPTM10 RPTM10 RPTM6b RPTM6b

RPTM11 RPTM11 N IZ N H H N N PTM-IIa PTM-IIa or or

RPTM6a RPTMsa RPTM5a RPTM6a RPTM8 RPTM8

RPTM7 RPTM7 O RPTM9 PTM9 N RPTM6 RPTM6 N

XPTM1 XpTM1 N RPTM10 RPTM6b

N RPTM11 RPTM11 IZ N H N H N

PTM-IIb PTM-Ilb wherein: wherein:

XPTMXPTM2,XPTM3,PTM4, and XPTM5,and XPTM1, XPTM2, XPTM3, XPTM4, XPTM5, XPTM6 XPTM6 areare independently independently selected selected from from CHN;or N; CH or

RPTM5a RPTM5a is is selectedfrom selected from the the group group consisting consisting of: bond, of: bond, optionally optionally substituted substituted amine, amine, optionally substituted optionally substituted amide amide (e.g., (e.g., optionally optionally substituted substituted with with an an methyl, alkyl, alkyl, methyl, ethyl, ethyl, RPTM5 RPTM5 0 o SS NH NH

propyl, ororbutyl o propyl, butylgroup), group), H, H, , -NHC(O)RPTM5; -NHC(O)RPTM5;

RPTM5 isisselected RPTM5 selected from the group from the group consisting of

145

Oct 2023 F F F F

N-----N-- N N

F F 2023248067 10

N----- N ----- F F ---- N

\N----- --- > -- --- - N N F

F F ----- F- F ) NN---- 0 ----- FW-0 N N----- FMW<

F F FF

Ho N ----- HON---- N N N Holl!!!!!

HOHO HO HO HO HON- HO F N

HO Ho RPTM6a and RPTM6b RPTM6a and RPTM6bare eachindependently are each selectedfrom independentlyselected hydrogen, fromhydrogen, halogen, or or halogen, optionally optionally

substitutedCl substituted C-C -C 6 alkyl alkyl (linear, (linear, branched, branched, optionally optionally substituted); substituted);

RPTM6 is isabsent, absent,hydrogen, halogen, hydrogen, aryl,aryl, halogen, methyl, ethyl, ethyl, methyl, OCH 3 , NHCH 3 or M1-CH OCH, NHCH 2-CH 2-M2, or M1-CH-CH-M2,

wherein M1 wherein M1isis CH, CHO2 , and 0 and NH, NH, and and M2 is M2 is hydrogen, hydrogen, alkyl, alkyl, cycliccyclic alkyl,alkyl, aryl aryl or or heterocycle; heterocycle;

RPTM7 is isabsent, absent,hydrogen, halogen, hydrogen, aryl,aryl, halogen, methyl, ethyl, ethyl, methyl, OCH 3 , NHCH 3 or M1-CH OCH, NHCH 2-CH 2-M2, or M1-CH-CH-M2,

wherein M1 wherein M1isisCH, CHO2 ,or or NH, 0 NH, and and M2 M2 is is hydrogen, hydrogen, alkyl, alkyl, cyclic cyclic alkyl, alkyl, arylaryl or or heterocycle; heterocycle;

RPTM8, RPTM9oror RPTM8, RPTM9 RPTM10 RPTMoare are independently independently selected selected fromgroup from the the consisting group consisting of of absent, absent, hydrogen, halogen,aryl, hydrogen, halogen, aryl, heteroaryl, heteroaryl, alkyl, alkyl, cycloalkyl, cycloalkyl, heterocycle, heterocycle, methyl, methyl, ethyl, ethyl, OCH OCH, 3 ,

NHCH NHCH or3 or M1-CH 2-CHwherein M1-CH-CH-M2, 2 -M2, wherein M1 is M1 is CH, O CH and2 ,NH, 0 and andNH, andhydrogen, M2 is M2 is hydrogen, alkyl, alkyl,

cyclic alkyl, aryl cyclic alkyl, aryl ororheterocycle; heterocycle; RPTM11 is absent, RPTMll is absent,hydrogen, hydrogen,halogen, methyl, halogen, ethyl, methyl, OCHOCH, ethyl, 3 , NH NH CH or M1-CH-CH-M2, CH 3or M1-CH 2-CH 2-M2, wherein M1 wherein M1isisCH, CHO2 ,or or NH, 0 NH, and and M2 M2 is is hydrogen, hydrogen, alkyl,alkyl, cyclic cyclic alkyl, alkyl, arylaryl or or heterocycle; heterocycle;

and and

146

Oct 2023 at at least least one of RPTM8, one of RPTM9or or RPTM8, RPTM9 RPTMO RPTM10 is modified is modified tocovalently to be be covalently joined joined to a aULM, a to a ULM,

chemical chemical linker linkergroup group(L), a CLM, (L), anan a CLM, ILM, a VLM, ILM, a VLM,MLM, MLM, aa ULM', ULM', aa CLM', CLM',a aILM', ILM',aa VLM', VLM', a aMLM', MLM', or combination or combination thereof. thereof.

[0270]

[0270] certain embodiments, In certain In thePTM embodiments, the PTMmay may comprise comprise a chemical a chemical group selected group selected from the from the 2023248067 10

group group ofof chemical chemical structures structures consisting consisting of: of:

RPTM5 RPTM5 0 o S SNH NH RPTM6a RPTM6a RPTM8 o RPTM8

0 RPTM7 RPTM7 XPTM3 PTM3 RPTM9 RPTM9 o RPTM6 XPB RPTM6 XPTM22 P 4 XPTM4

XPTM1 XPTM1 XPTM5 XPTM5 XPTM6 RPTM10 'RPTM10 RPTM6b RPTM6b RPTMM

RPTM11 RPTM11 ZI N H N N o orr

RPTM5 RPTM5 O o \/1 S NH 1 Is e biNHeRPTMia RPTM6a o RPTms RPTM8

w RPTM7 RPTM9 h O RPTus RPTM6 N

XPTM1 N RPTM10 RPTM6b RPTM1 TRPTMe N) -- RPTM11 1 RPTM~ XPT-11,

1 IrZI bwR RPTM11 N N

wherein RPTM5, wherein RPTM5,RPTM6a, RPTM6a, RPTM6b, RPTM6b, RPTM6, RPTM6,RPTM7, RPTM7,RPTM8, RPTM8,RPTM9, RPTM9,RPTM10, RPTM10,RPTM11 RPTMlliare described areasasdescribed

herein. herein.

[0271] In some

[0271] In some embodiments, embodiments, whenisRPTM9 when RPTM9 is the covalently the covalently position, joined position, RPTM7 and and RPTM8 RPTM8 RPTM7 joined can be connected can be connectedtogether togethervia via aa covalent covalent bond bondininaaway waytotoform forma abicyclic bicyclicgroup groupwith the with thering ringtoto which RPTM7 which andRPTM8 RPTM7and RPTM8 areare attached. attached.

[0272] In In

[0272] other other embodiments, whenwhen embodiments, RPTM8RPTM8 is the the covalently is covalently joined joined position,RPTM9 position, andand RPTM9

RPTM10 RPTM10 can be connected can be togethervia connectedtogether viaa acovalent bondinina away covalentbond wayto to form form a bicyclic group a bicyclicgroup with with thethe

RPTad to which ring to ring which RPTM9 andRPTM10 RPTM9 and RPTM10are attached. areattached.

147

In further

[0273] In further

[0273] embodiments, embodiments, when is when RPTM10 is the covalently the covalently RPTMO joined position, RPTM8 andRPTM8 joined position, and canbe RPTM9 can RPTM9 beconnected connectedtogether togethervia viaa acovalent covalentbond bond in in a way a way to to form form a bicyclic a bicyclic group group with with the the

ring to ring to which which RPTM8 andRPTM9 RPTM8 and areattached. RPTM9are attached. anyany

[0274] In In

[0274] aspect aspect embodiment ororembodiment describedherein, described PTM the PTM herein, the may may comprise comprise a chemical a chemical

group selected from group selected from the the group group of ofchemical chemicalstructures structures consisting consisting of of PTM-III: PTM-III:

XPTM16 XPTM16 O O XPTM15 RPTM13

RpTM14 RPTM14 PTM10 XPTM10 RPTM12 RPTM12 H ZI H IIIIIIIII XPTM17 XPTM17 RPTM17 RPTM17 XP \lTM XPTM11 XPTM9 RPTM18 N \ o

XP XPTM20 XPTM14 XPTM1 4 XPTM8 XPTM8 // \ N N XPTM12 XPTM7 RPTM16 RPTM16 XPTM19 XPTM19 11

XPTM18 XPTM13 XPTM13 N.-O RPTM15 RPTM19 XPTM18 RpTI9PTI8N N 0 ZI N N H O H RPTM20 RPTM20 RPTM21 RPTM21

PTM-III PTM-III

wherein: wherein:

XPTM7, XPTM8, XPTM7, XPTM8,XPTM9, XPTM10, XPTM9, XPTM11, XPTM1O, XPTM12, XPTM11, XPTM13, XPTM12, XPTM14,XPTM14, XPTM13, XPTM15, XPTM15, XPTM16, XPTM17, XPTM16, XPTM17,

XPTM18, XPTM19, XPTM20 XPTM18, XPTM19, XPTM2 are or or independentlyCHCH are independently N;N;

RPTM12, RPTM13, RPTM12, RPTM13, RPTM14, RPTM15,RPTM16, RPTM14, RPTM15, RPTM17, RPTM16,RPTM17, RPTM18, RPTM18, RPTM19 are are RPTM19 independently independently selected selected

from the group from the groupconsisting consisting of of absent, absent, hydrogen, hydrogen, halogen, halogen, aryl, aryl, heteroaryl, heteroaryl, cycloalkyl, cycloalkyl,

heterocycle, heterocycle,methyl, methyl,ethyl, otherother ethyl, alkyl,alkyl, OCH 3 ,OCH, NHCH 3 oror NHCH M1-CH 2-CH 2 -M2, M1-CH-CH-M2, wherein wherein M1M1

is is CH CH,2 ,O0and andNH, NH,andand M2 M2 is hydrogen, is hydrogen, alkyl, alkyl, cyclic cyclic alkyl,aryl alkyl, arylororheterocycle; heterocycle; RPTM20 isis RPTM20 a small small group group containing less less than than four four non-hydrogen atoms; non-hydrogen atoms;

RPTM21 is selectedfrom RPTM21is selected from the the group group consisting consisting of trifluoromethyl, of trifluoromethyl, chloro,fluoro, chloro, bromo, bromo, fluoro, methyl, methyl, ethyl, propyl,isopropyl, ethyl, propyl, isopropyl, tert-butyl, tert-butyl, butyl, butyl, iso-butyl, iso-butyl, cyclopropyl, cyclopropyl, cyclobutyl, cyclobutyl, cyclopentyl, cyclopentyl,

cyclohexyl, cyclohexyl, OCH OCH,3 , NHCH, NHCHdimethylamino 3 , dimethylamino or or M1-CH 2-CHwherein M1-CH-CH-M2, 2 -M2, wherein M1 Ois CH2 , 0 M1 is CH,

or NH,andand or NH, M2 M2 is hydrogen, is hydrogen, alkyl, alkyl, cyclic cyclic alkyl, alkyl, aryl oraryl or heterocycle; heterocycle; and and at at least least one one of of RPTM12, RPTM12, RPTM13 andRPTM16 RPTM13 and RPTM16is is modifiedtotobebe covalently modified joined covalently joined to a to ULM,a ULM, a a chemical chemical linker linkergroup group(L), a CLM, (L), anan a CLM, ILM, a VLM, ILM, a VLM,MLM, MLM, aa ULM', ULM', aa CLM', CLM',a aILM', ILM',aa VLM',a aMLM', VLM', MLM', or combination or combination thereof. thereof.

148

2023248067 10 2023

[0275] In In

[0275] some some embodiments, whenwhen embodiments, RPTM12 is the RPTM12 the covalently is covalently joined joined position,RPTM13 position, and RPTM13and

can be RPTM14 can RPTM14 connectedtogether be connected viaa acovalent togethervia bondinina away covalentbond wayto to form form a bicyclic group a bicyclicgroup with with thethe

Oct ring to ring to which whichRPTM13 andand RPTM13 RPTM14 RPTM14 are attached; are attached; and/or and/or RPTM15RPTM15 and can and RPTM16 RPTM16 can be connected be connected

together together via via aa covalent covalent bond in aa way bond in to form way to form aa bicyclic bicyclic group group with with the the ring ring to to which which RPTM15 and RPTM15and

areattached. RPTM16 are RPTM16 attached.

[0276] In In

[0276] other other embodiments, embodiments, when when RPTM13 RPTM13 is the is the covalentlyjoined covalently position, RPTM12 joinedposition, and RPTM12 and

can be RPTM16 can RPTM16 be connected connectedtogether togethervia viaa acovalent covalentbond bondinina away wayto to form form a bicyclicgroup a bicyclic group with with thethe

ring to ring to which whichRPTM12 andand RPTM12 RPTM16 RPTM16 are attached; are attached; and/or and/or RPTM15RPTM15 and can and RPTM16 RPTM16 can be connected be connected

together via aa covalent together covalent bond in a way bond in to form way to form aa bicyclic bicyclic group group with with the the ring ring to to which which RPTM15 and RPTM15and

RPTM16 areattached. RPTM16are attached.

[0277] In further

[0277] In further embodiments, embodiments, when is when RPTM16 is the covalently the covalently RPTM16 joined position, joined position, RPTM12 and RPTM12 and can be RPTM13 can RPTM13 be connected connectedtogether togethervia viaa acovalent covalentbond bondinina away wayto to form form a bicyclicgroup a bicyclic group with with thethe

ring to ring to which whichRPTM12 andand RPTM12 RPTM13 RPTM13 are attached; are attached; and/or and/or RPTM13RPTM13 and can and RPTM14 RPTM14 can be connected be connected

together via together via aa covalent covalent bond in aa way bond in to form way to form aa bicyclic bicyclic group group with with the the ring ring to to which which RPTM13 and RPTM13and

areattached. RPTM14 are RPTM14 attached. anyany

[0278] In In

[0278] ororembodiment aspect aspect embodiment described the PTM herein, the describedherein, PTM may may comprise comprise a chemical a chemical

group selected from group selected from the the group group of ofchemical chemicalstructures structures consisting consisting of of PTM-IVa PTM-IVa or or PTM-IVb: PTM-IVb:

RPTM22 RPTM22 o S NH RPTM25a NH RPTM25a RPTM26 RPTM26 N N o RPTM25 RPTM25 XM2 XPTM29

XPTM21 XPTM21 XPTM23 XPTM 23 / N N XPTM27 XPTM27 N N XPTM28 XPTM28 RPTM28 XPT 22/ XPTM22

RPTM27 RPTM27 RPTM30PRPTM29 RpTM25b RPTM25b XPTM24 XPTM26 RPTM30 XPTM26 XPTM24 XPTM34 XPTM33 XPTM34=XPTM33 XPTM25 XPTM25 - RPTM24 N XPTM32 XPTM32 N /N// XPTM31 RPTM RPTM23 XPTM30-XPTM31 XPTM30

RPTM32 RPTM32 RPTM3 RPTM31

PTM-IVa PTM-IVa or or

149

RPTM22 RPTM22 0 o \//° S /S N NH RPTM26 RPTM25a RPTM26 N NH RPTM25a N o RPTM25 RPTM25 XPTM29 XPTM2

XPTM21 XPTM214 XPTM23 XPT31 XPTM27 --'XPTM27 \/ N N N N" XPTM28 XPTM28 N, RPTM28 RPTM28 XPTM22/ XPTM22 2023248067

RPTM27 RPTM27 RPTM29 RPMT25b RPMT25b XPTM24 XPTM26 RPTM30 RPTM30 RPTM29 XPTM26 XPTM24 XPTM34 XPTM33 XPTM34-XPTM33 XPTM25 XPTM25 RPTM24 RPTM24 XPTM32 XPTM32 N N XPTM30 XPTM31 RPTM23 RPTM23 XPTM 3 0 XPTM31

RPTM32 RPTM32 RPTM RPTM31

PTM-IVb PTM-IVb ,

wherein: wherein:

XPTM21, XPTM21, XPTM22, XPTM22,XPTM23, XPTM23,XPTM24, XPTM25, XPTM24, XPTM26, XPTM25, XPTM27, XPTM26, XPTM28, XPTM27, XPTM29, XPTM28, XPTM30, XPTM29, XPTM31,XPTM31, XPTM30,

XPTM32, XPTM33, XPTM32, XPTM33, XPTM34 XPTM34 are or or independentlyCHCH are independently N;N;

selected from RPTM22 isisselected RPTM22 from the group consisting the group consisting of of

150

F F F F N-----N-- N N F F

N ON----- ----- F4* F NN----

\N----- --- > -- --- - N N F

----- F N----- F ----- F N---- F N F F N F F FF

HO N ----- HON---- N N N HOHO HO HO HO Ho HOF- Ho F N

HO Ho RPTM25a and RPTM25b RPTM25aand RPTM25bare are each independentlyselected each independently fromhydrogen, selectedfrom or or halogen, hydrogen,halogen, C-C -C 6 alkyl C1alkyl (linear, (linear, branched, optionally branched, optionally substituted); substituted);

RPTM23, RPTM24, RPTM28, RPTM29, RPTM30, RPTM24,RPTM28,RPTM29, RPTM31, RPTM32 are independently selected M30,RRPTM31,RPTM32are independently fromthe selected from the group consisting group consisting of of absent, absent, bond, bond, hydrogen, hydrogen, halogen, halogen, aryl (optionally aryl (optionally substituted), substituted),

heteroaryl(optionally heteroaryl (optionally substituted), substituted), cycloalkyl cycloalkyl (optionally (optionally substituted), substituted), heterocycle heterocycle

(optionallysubstituted), (optionally substituted),methyl, methyl, ethyl ethyl (optionally (optionally substituted), substituted), other other alkyl (linear, alkyl (linear,

branched, optionally branched, optionallysubstituted), OCHOCH, substituted), 3 , NHCH NHCH3 or orM1-CH 2 -CH 2-M2, M1-CH-CH-M2, wherein wherein M1 M1 is is CH CH, 2O, 0 and and NH, NH, and and M2hydrogen, M2 is is hydrogen, alkylalkyl (linear, (linear, branched, branched, optionally optionally substituted), substituted),

cyclic alkyl(optionally cyclic alkyl (optionallysubstituted), substituted), aryl aryl (optionally (optionally substituted)or substituted)or heterocycle heterocycle

(optionally substituted);andand (optionally substituted);

RPTM25 is absent, RPTM25 is absent, hydrogen, hydrogen,halogen, halogen,C-C C1 alkyl -C6 alkyl (linear, (linear, branched, branched, optionally optionally substituted), substituted),

OCH3, OCH3, NHCH or SCH NHCH 3or SCH;3 ; RPTM26 is absent, RPTM26 is absent, hydrogen, hydrogen,halogen, halogen,C-C C 1alkyl -C 6 alkyl (linear, (linear, branched, branched, optionally optionally substituted), substituted),

OCH3, OCH3, NHCH or SCH NHCH 3or SCH;3 ;

151

RPTM27 isisselected selected from from the group consisting of 2023248067 10 Oct 2023

RPTM27 group consisting of absent, absent, hydrogen, halogen, C-C hydrogen, halogen, Cl-C 6 alkyl (linear, alkyl (linear,

branched, optionally branched, optionallysubstituted), OCHOCH, substituted), 3 , NHCH NHCH3 or or SCH 3 ; and SCH; and at at least least one of RPTM24, one of RPTM24, RPTM29, RPTM29,RPTM32 is is RPTM32 modified modified tob to be e covalently covalently joined joined to a aULM, a to a ULM,

chemical chemical linker linkergroup group(L), a CLM, (L), anan a CLM, ILM, a VLM, ILM, a VLM,MLM, MLM, aa ULM', ULM', aa CLM', CLM',a aILM', ILM',aa VLM',a aMLM', VLM', MLM', or combination or combination thereof. thereof.

[0279] In In

[0279] some some embodiments, whenwhen embodiments, RPTM24 is covalently is the RPTM24 the covalently joined joined position,RPTM31 position, and RPTM31 and

RPTM32 can be RPTM32 can be connected connectedtogether togethervia viaa acovalent covalentbond bondinina away wayto to form form a bicyclicgroup a bicyclic group with with thethe

ring to ring to which andRPTM32 RPTM31 and which RPTM31 areare RPTM32 attached;or orRPTM29 attached; andand RPTM29 RPTM30 canconnected can be RPTM30 be connected together together

via a covalent bond in aa way bond in waytoto form forma abicyclic bicyclic group groupwith with the ringtotowhich the ring whichRPTM29 andand RPTM29 RPTM30 RPTM30

are attached. are attached.

[0280] In In

[0280] other other embodiments, embodiments, when when RPTM29 is the is the RPTM29 covalentlyjoined covalently position, RPTM24 joinedposition, and RPTM24 and

RPTM32 can be RPTM32 can be connected connectedtogether togethervia viaa acovalent covalentbond bondinina away wayto to form form a bicyclicgroup a bicyclic group with with thethe

ring to ring to which whichRPTM24 andand RPTM24 RPTM32 RPTM32 are attached; are attached; and/or and/or RPTM31RPTM31 and can and RPTM32 RPTM32 can be connected be connected

together via together via aa covalent covalent bond in aa way bond in to form way to form aa bicyclic bicyclic group group with with the the ring ring to to which which RPTM31 and RPTM31 and

areattached. RPTM32 are RPTM32 attached.

[0281] In further

[0281] In further embodiments, embodiments, when is when RPTM32 is the covalently the covalently RPTM32 joined position, joined position, RPTM24 and RPTM24 and can be RPTM29 can RPTM29 be connected connectedtogether togethervia viaa acovalent covalentbond bondinina away wayto to form form a bicyclicgroup a bicyclic group with with thethe

ring to ring to which whichRPTM24 andand RPTM24 RPTM29 RPTM29 are attached; are attached; and/or and/or RPTM29RPTM29 and can and RPTM30 RPTM30 can be connected be connected

together via aa covalent together covalent bond in a way bond in to form way to form aa bicyclic bicyclic group group with with the the ring ring to to which which RPTM29 and RPTM29and

areattached. RPTM30 are RPTM3o attached.

[0282] In any

[0282] In aspect any aspect or embodiments or embodiments described described herein, herein, the PTMthe is PTM is selected selected from thefrom groupthe group consisting consistingofofchemical structures chemical PTM-1, structures PTM-2, PTM-1, PTM-3, PTM-2, PTM-4, PTM-3, PTM-4,PTM-5, PTM-5, PTM-6, PTM-6, PTM-7, and PTM-7, and

PTM-8: PTM-8:

152

F 1 F 0, N S N NH F HO' N,'I N O F HO-N O NN

F N N IZ N N N H 2023248067 PTM-1 PTM-2 PTM-2 O===O O 0 O 10 S NH F O N O O NH IZ F FN N N 0 o F- F H ZI N H N F H FF PTM-3 PTM-3 PTM-4 PTM-4

\N N O===O 0 o o o ZI F OH F ' / N S NH ,NH S N 0 F

N O N N F N N F IZ N N N H PTM-5 PTM-5 T PTM-6 PTM-6 N N ZI 0 0 0 N"H N FN ~ o SHN F o oH 0/H 'N F S N N N N~ N N N N NII F N N N PTM-7 PTM-7 PTM-8 PTM-8

153

Oct 2023 F F O O N S F NO NH F F F: N NO' S NH F O F O O N C'\0 N O N C c 2023248067 10

F F F F CN N

IZ IZ N H NH N N N H H H N PTM-9 PTM-9 PTM-10 PTM-10

[0283]

[0283] XI. Compounds XI. Targeting FKBP: Compounds Targeting FKBP: MeO MeO

MeO H ZI H O N N'R R O N The 0 O O

MeO MeO OMe OMe

[0284]

[0284] OMe

[0285] (Derivatized

[0285] (Derivatized wherewhere "R" designates "R" designates a site afor sitea for a linker linker groupgroup L or aL -(L-CLM) or a -(L-CLM) group group for example). attachment, for example).

[0286] XII.

[0286] XII.Compounds Compounds TargetingAndrogen Targeting AndrogenReceptor Receptor (AR) (AR)

[02871 1. RU59063

[0287] 1. RU59063 Ligand Ligand (derivatized) (derivatized) of of Androgen Androgen Rceptor Rceptor

NCa NC S S F3C N N FC '

N 0/ O O

[0288]

[0288] R

[0289] (Derivatized

[0289] (Derivatized wherewhere "R" designates "R" designates a site afor sitea for a linker linker groupgroup L or aL -(L-CLM) or a -(L-CLM) group group attachment, for for example). example).

[0290] 2. 2.

[0290] SARMSARM Ligand Ligand (derivatized) (derivatized) of of Androgen Androgen Receptor Receptor

154

Oct 2023 F3C FC O O 0 2N N N - ON H H -NHH \ / NH N OH O R R

[02911 0 2023248067 10

[0291] O

[0292] (Derivatized

[0292] (Derivatized wherewhere "R" designates "R" designates a site afor sitea linker for a linker group group L or a-(L-CLM) L or a-(L-CLM) group group attachment, for example). attachment, for example).

[0293]

[0293] 3. Androgen 3. Androgen Receptor Receptor Ligand Ligand DHT DHT (derivatized) (derivatized)

0 O O N-R ZI R N H H

[0294]

[0294] O

[0295] (Derivatized

[0295] (Derivatizedwhere where"R""R" designatesa asite designates site for fora alinker group linker L or group L or-(L-CLM) group -(L-CLM) group

attachment, for example). attachment, for example).

[0296]

[0296] 4. MDV3100 4. MDV3100Ligand(derivatized) Ligand (derivatized)

5s - R S NC/\ NC N N 1 N N F 3C FC

[02971

[0297] O

[0298]

[0298] 5. ARN-509 5. ARN-509 Ligand Ligand (derivatized) (derivatized)

R N S NC NC N N N R N F FC3C

[0299]

[0299] 0 O

[0300]

[0300] 6. Hexahydrobenzisoxazoles 6. Hexahydrobenzisoxazoles

N N-X RR F3C FC

[0301]

[0301] NC

[0302]

[0302] 7. Tetramethylcyclobutanes 7. Tetramethylcyclobutanes

155

2023 R R

2023248067 10 Oct C1 CI O O "N N 0 O

[03031

[0303] NC NC H

[0304]

[0304] 8. In 8. In any aspect or any aspect or embodiment described embodiment described herein,thethePTM herein, PTM is aischemical a chemical moiety moiety that that

binds to binds to the the androgen receptor (AR). androgen receptor (AR).Various Variousandrogen androgen receptor receptor binding binding compounds compounds have have been been described in literature, described in literature, including including various androgen derivatives various androgen derivatives suchas astestosterone, such testosterone, dihydrotestosterone, andmetribolone dihydrotestosterone, and metribolone(also (also known known as methyltrienolone as methyltrienolone or R1881), or R1881), and non-and non

steroidal steroidal compounds such compounds such as as bicalutamide, bicalutamide, enzalutamide, enzalutamide, somesome of which of which are described are described above. above.

Those Those ofofordinary ordinaryskill skillin in thethe art art would would appreciate appreciate that these that these androgen androgen receptorreceptor binding binding

compounds couldbebepotentially compounds could potentially used used as as an an androgen androgen binding binding moiety moiety (ABM) in aa PROTAC (ABM) in PROTAC compound. Such compound. Such literatureincludes, literature includes,butbutnotnot limited limited to,to, G. G. F. F. Allan Allan et. et. al,al, Nuclear Nuclear Receptor Receptor

Signaling, 2003, Signaling, 2003, 1,1, e009; e009;R.R.H.H.Bradbury Bradbury et. al, et. al, Bioorganic Bioorganic & Medicinal & Medicinal Chemistry Chemistry Letters,Letters,

2011 5442-5445; 2011 5442-5445;C. C. GuoGuo et. et. al,al,Bioorganic Bioorganic & Medicinal & Medicinal Chemistry Chemistry Letters, Letters, 2012 2012 2572-2578; 2572-2578; P. P. K. Poutiainen K. Poutiainen et. et. al, al,JJ.Med. Med. Chem. 2012,55, Chem. 2012, 6316- -6327 55, 6316 6327 A. A. Pepe Pepe et. et. al,al, J.JMed. Med.Chem. Chem. 2013, 2013,

56, 8280 -- 8297; 56, 8280 8297;M.M.E.E.Jung Jungetetal, al, J. J Med. Med.Chem. Chem. 2010, 2010, 53,53, 2779-2796, 2779-2796, which which are incorporated are incorporated

by reference by reference herein herein In any

[0305] In any

[0305] aspect aspect or embodiment or embodiment described described herein, herein, the ABM ABM comprises thecomprises a structure a structure

selected from,butbut selected from, notnot limited limited to the to the structures structures shownshown below, below, wherein wherein a dashed a dashed line linetheindicates the indicates

attachment point of attachment point of aa linker linker moiety moiety or or aa ULM, suchasasaaCLM: ULM, such CLM: Y² R1 Y2 R¹ R²

w¹ N _R2 N N Y N ( W² Y¹ )

ABM-a ABM-a ;

(RQ) 0 6 (R²)- Y5 Y3 Y³ Q Y, 4 W² Q .

Y w1

ABM-b ABM-b

156

Oct 2023 y1 Y¹ 1 ga y3 Y3 Y ,R~ w2 W W² W¹ N R Rb Y Y² R 2023248067 10

ABM-c ABM-c ;and ; and

Y³ W² WW2 (Y³)-5 Q w1

ABM-d ABM-d

wherein: wherein:

W' W¹ isisaryl, aryl, heteroaryl, heteroaryl,bicyclic, bicyclic,or or biheterocyclic, biheterocyclic, each each independently independently substituted substituted by 1 or by 1 or

moreH,H,halo, more halo, hydroxyl, hydroxyl, nitro, nitro, CN, C=CH,C CN, C=CH, C1-61 -6alkyl (linear, branched, alkyl (linear, branched, optionally optionally substituted; for substituted; forexample, example, optionally optionally substituted substitutedby by1 1orormore more halo,C1.6alkoxyl),Ci-6 halo, C- alkoxyl), C-

alkoxyl (linear, branched, optionally substituted; for example, optionally substituted by 1 alkoxyl (linear, branched, optionally substituted; for example, optionally substituted by 1

or morehalo), or more halo),C2-6alkenyl,C2-6alkynyl, or CF 3; C2-6 alkenyl, C2-6 alkynyl, or CF;

Y¹, Y²2 are Yl, Y are each NR 1O, independently NRY¹, each independently , 0,S;S; Y Y³,, y , y 5 areeach 3 Y,4 Y are eachindependently independently a bond, a bond, O, 0, NRY², NRY 2 CRY¹RY², 2 C=S, SO, SO, C=O, , CR'RY , C=0, C=S, SO, SO 2

, heteroaryl, or aryl; heteroaryl, or aryl;

Q is aa 3-6 Q is 3-6 membered ringwith membered ring with0-4 0-4heteroatoms, heteroatoms,optionally optionallysubstituted substitutedwith with0-6 0-6R°, RQ,each each RQ,is R°, isindependently independently H, C1 -6alkyl H, C- (linear, branched, alkyl (linear, branched, optionally optionally substituted; for example, substituted; for example,

optionally substituted optionally substituted by by 1 or1 more or more halo,halo,Ci-6alkoxyl), halogen,Ci-6alkoxy, C1-6 alkoxyl), halogen, C1-6 alkoxy, or 2 R° or 2 RQ

groups taken together groups taken together with with the the atom atom they they are are attached attached to, to, form form a a 3-8 3-8 membered ring membered ring

system containing 0-2 system containing 0-2 heteroatoms); heteroatoms); R', R R¹, 2 R², R, R, , Ra, R ,RRy2are R,R¹¹, eachindependently ² are each independentlyH,H,C- C1-6alkyl (linear, alkyl (linear, branched, branched, optionally optionally

substituted; for substituted; forexample, example, optionally optionally substituted substitutedby by1 1orormore morehalo,C1-6alkoxyl), halo, C- alkoxyl), halogen, halogen,

CI-6alkoxy, cyclic, heterocyclic, or R1 , R2 together C- alkoxy, cyclic, heterocyclic, or R¹, R² together with the atom they are attached to, with the atom they are attached to, form form aa 3-8 3-8 membered membered ringsystem ring system containing containing 0-20-2 heteroatoms); heteroatoms);

W² 2isisa bond, C- alkyl, C- heteroalkyl, O, aryl, heteroaryl, alicyclic, heterocyclic, W a bond,C1 -6alkyl,C1 -6heteroalkyl, 0, aryl, heteroaryl, alicyclic, heterocyclic, biheterocyclic, biaryl, or biheteroaryl, each optionally substituted by 1-10 Rw 2; biheterocyclic, biaryl, or biheteroaryl, each optionally substituted by 1-10 R²;

157

each R2 each RW² is isindependently independently H, halo, H, halo, C 1-6 C- alkyl alkyl (linear, (linear, branched, branched, optionallyoptionally substituted; substituted; for for example, optionally substituted example, optionally substituted by by 1 1 or more F), 2 , C- cycloalkyl, C4-6 -ORW²A or more F), -ORw A ,C3-6 cycloalkyl, C4 -6

cycloheteroalkyl, cycloheteroalkyl, C- C 1-6 alicyclic alicyclic (optionally (optionally substituted), substituted), heterocyclic heterocyclic (optionally (optionally

substituted), aryl(optionally substituted), aryl (optionallysubstituted), substituted), or heteroaryl or heteroaryl (optionally (optionally substituted), substituted), bicyclic bicyclic

2 hereoaryl or aryl, 2 , NRY Rv , CN; and hereoaryl or aryl,OCi-3alkyl (optionallysubstituted), OC-alkyl (optionally substituted), OH, NHNR OH, NH, Y¹RY², CN; and

RW2 Aisis H, RW²A H, C- C 1-6 alkyl(linear, alkyl (linear, branched), branched), or or C- C 1-6heteroalkyl(linear, heteroalkyl (linear, branched), branched), each each optionally optionally substituted bya acycloalkyl, cycloalkyl, cycloheteroalkyl, aryl, aryl, heterocyclic, heteroaryl, halo, orhalo, OC1- or OC1 2023248067

substituted by cycloheteroalkyl, heterocyclic, heteroaryl,

. 3alkyl. alkyl.

2

[0306]

[0306] In any In any aspect or embodiment aspect or embodiment described describedherein, theW²Wis herein,the iscovalently oneoror coupledtotoone covalentlycoupled more ULM more ULM or CLM or CLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as described herein. described herein.

(R23)0-4 (R)-4 MVV

R22

[0307]

[0307] In any In any aspect or embodiment aspect or embodiment described describedherein, 1 is herein,W¹Wis - R or or

(R23)0-3 (R)- MVV

R22

N R , wherein each R 2 isindependently , wherein each R 2is independentlyhalo, halo,H,H,optionally optionallysubstituted substitutedalkyl, alkyl, haloalkyl, cyano, haloalkyl, cyano, or nitro; nitro;and andeach eachRR2 3is is independently independentlyH,H,halo, halo, CF, CF 3optionally , optionallysubstituted substituted alkyl, alkoxy,haloalkyl, alkyl, alkoxy, cyano, haloalkyl, cyano, or or nitro. nitro.

[0308]

[0308] In any aspect In any or embodiment aspect or embodiment described describedherein, 1 isselected herein,W¹Wis selectedfrom fromthe thegroup group consisting of: consisting of:

CF CF F F CI CI CF3 CF3 ^^^^ MVV MVV MVV MVV

1_ CN. CN 1-0 -NO 2 . / CN CN J \/ CN. CN CN CN ; NO ; ; ;

CI CI CI OMe F F C1 CI { C1 CF 3 CF OMe ~~~~

CN ~ CN s / CN { CN { ~ CN MCN CN CN F N CN F N N ;and ; N ; and

CN CN

158

ABM comprises thecomprises a structure 2023248067 10 Oct 2023

[0309] In any

[0309] In any aspect aspect or embodiment or embodiment described described herein, herein, the ABM a structure

rrr selected selected from the following from the structures shown following structures below,where shown below, wherea a indicates tha indicates tha attachment attachment

point of point of aa linker linkerorora a ULM: ULM:

N 2 N N R° RQ3 3 R°. R~ Y4 N N Y5Y5

R°2 Y³ R° R°R RQ3 R R

R2 y3 R R° 2 RQ3 R° R R Y4 N Y 0 R R°2 R Y³ R R° R° RQ and , and

Ro2 y3 RR' R° R03 RQ3 R°. ROY4 Y4 K \ RQ Y Y³ R° R R°

wherein: wherein:

R² is a H, R2is H, halogen, halogen,CH CH3 or or CF 3; CF; 3 RQ is R³ is H, H, halo, halo, hydroxyl, hydroxyl, nitro, nitro, CN, C--CH,C- Calkyl CN, C=CH, 1-6 alkyl (linear, branched, optionally (linear, branched, optionally

substituted substituted by by 11 or or more more halo, halo, C C-1-6alkoxyl), C- alkoxyl), C 1alkoxyl -6 alkoxyl (linear, branched, (linear, branched,optionally optionally substituted substituted by by 11 or or more more halo), halo), C2-6 C2-6alkenyl, alkenyl,C2-6 C- alkynyl, alkynyl, or or CF CF;3 ;

Y3 , Y Y³, 5 are Y,4 YYare eachindependently each independently a bond, a bond, O, 0, NRY NRY², 2 , CRYIRv CRY¹R², C=O, heteroaryl, C=0,2 ,heteroaryl, or or aryl; aryl; R 1, RRY2 R¹¹, are each ² are each independently independentlyH,H,ororC- C 1alkyl -6 alkyl (linear, branched, (linear, branched,optionally optionally substituted substituted by by 11 or or more halo, C more halo, C-1-6 alkoxyl,cyclic, alkoxyl, cyclic, or or heterocyclic); heterocyclic); and and

159

RQ each R° each independently independentlyisisH, H,C-C alkyl alkyl (linear,branched, (linear, branched,optionally optionallysubstituted substitutedbyby1 1oror 2023248067 10 Oct 2023

CI-C 6

more halo, more halo, or or C- C 1 .6alkoxyl), alkoxyl),orortwo twoR°RQtogether togetherwith withthe theatom atomthey theyareareattached attachedto, to, form forma a 3-8 3-8 membered ringsystem membered ring system containing containing 0-20-2 heteroatoms. heteroatoms.

[0310]

[0310] In any In any aspect or embodiment aspect or embodiment described describedherein, herein,each eachR°RQ is isindependently H or independently CH.CH H or In . In 3

another another embodiment R³Q3is embodiment R is CN.

[0311]

[0311] In In any any aspect or embodiment aspect or embodiment described theABM herein,the describedherein, ABM comprises comprises a structure a structure

selected selected from the following from the structures shown following structures below,where shown below, wherea a indicates the indicates the attachment attachment

point of point of aa linker linkerorora a ULM: ULM:

N N N N N

4 RQ RQ3 3 Y Y4 Rs Yy y5 Y5 N N

R°2 Y³ N N 2 Rs RQ3 Y4 y4 5 Y Y5 N N

RQ2 R°2 y3"I Y³

x X R0 4 y X 2 Q3 Y4 R X Y5 X

Y³ , and and R Ros RQ3 Y4 y4 ysk Y5

RRQ2 ² y3a Y³

wherein: wherein:

RQ2 is aa H, R² is H, halogen, halogen,CN, CN,CH or CF CH3 or CF;3 ; and

160

3 H, halo, hydroxyl, nitro, CN, C=CH, C- alkyl (linear, branched, optionally Oct 2023 RQis R³ is H, halo, hydroxyl, nitro, CN, C-CH, C 1-6 alkyl (linear, branched, optionally substituted substituted by by 11 or or more more halo, halo, C C-1-6 C 1alkoxyl alkoxyl), C- alkoxyl), -6 alkoxyl (linear,branched, (linear, branched,optionally optionally substituted substituted by by 11 or or more more halo), halo), C2-6 alkenyl, C- C- alkenyl, C2-6 alkynyl,ororCF; alkynyl, CF.3 ;

. Y3, Y Y³, 5 are Y,4 YYare eachindependently each independently a bond, a bond, O, 0, NRY NRY², 2 C=O,2 ,heteroaryl, , CRYIRv CRY¹R², C=O, heteroaryl, or and or aryl; aryl; and 2023248067 10

R 1, RRY2 R¹¹, are each ² are each independently independentlyHHororC-C alkyl 1-6 alkyl (linear, branched, optionally substituted by 1 (linear, branched, optionally substituted by 1

or more halo, C more halo, C-1-6 alkoxyl,cyclic, alkoxyl, cyclic, or or heterocyclic); heterocyclic); and and

X is X is N or C. N or C.

[0312]

[0312] In any In any aspect or embodiment aspect or embodiment described describedherein, is3 is herein,R³RQ a CN. a CN.

[0313]

[0313] In In any aspect or any aspect or embodiment embodiment described theABM herein,the describedherein, ABM comprises comprises a structure a structure shown shown

below, where below, wherea adashed dashedline lineindicates indicates the the attachment attachment point point of of aa linker linker moiety or aa ULM moiety or ULM orora aCLM: CLM:

R¹ R² R¹ R² W² (Y) (R )12 R¹ R1 4 Y3 R¹

your R

R¹ R1 R² R2 R² R2 R¹ R1

W¹

wherein: wherein:

(R23)-3 (R23)0-3 (R 2 3) 0-4 (R)-4 MVV

N MVV

-<')\-R2 R22 N R W¹ is W,

each is

each RR 2is R R2or or

is independently H or -CN; independently H or -CN; ;

2

each each RR 2is 3 is independentlyH, H, independently halo,C-CCI-C halo, 6 alkyl alkyl (linear, (linear, branched, branched, optionally optionally substituted),C-C1 substituted),

C alkoxy, or C 6alkoxy, or -CF3; -CF; Y³ 3is Y is aa bond bondororO;0; Y 4isis aa bond Y bondoror NH; NH; Y 5isis aa bond, Y bond, C=0, C=0,C1-C C1 -C6 heteroaryl, or C1 -C6 aryl; heteroaryl, or C-C aryl;

R', R 2 , are are each each independently H, or or C-C 6 alkyl (linear or branched, optionally substituted; R¹, R², independently H, CI-Calkyl (linear or branched, optionally substituted;

for for example, optionally substituted example, optionally substituted by by 11 or or more more halo, halo, or or CC- 1-6 alkoxyl); alkoxyl);

W² 2is W is aa bond, bond, C C- -6 aryl, aryl, C- C 1heteroaryl, -6 heteroaryl,C-Calicyclic, or C- heterocyclic, biheterocyclic, 1 1-6 alicyclic, or C 1- 6 heterocyclic, biheterocyclic,

biaryl, or biaryl, or biheteroaryl, biheteroaryl,each each optionally optionally substituted substituted R²; andRw 2 ; by 1-10 by 1-10 and

161

Oct 2023 each each R²² R2isis independently independently H, H,or or halo; halo; and and ~ represents representsa abond bond that that maymay be stereospecific be stereospecific ((R) or((R) (S))or or(S)) or non-stereospecific. non-stereospecific.

2

[0314]

[0314] In any aspect In any or embodiment aspect described or embodiment herein, described the W²the herein, is W is covalently covalently coupled to onetoorone coupled or more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as 2023248067 10

described herein. described herein.

[0315]

[0315] In any aspect In any or embodiment aspect described or embodiment herein, described W¹ is W herein, 1 is selected selected from the fromgroup the group consisting of: consisting of:

F CI1CI CI CF 3 CF 3 CF F CF $ {

CN MVV \/ //// CN MVV

CN. 1 CN. CN CN CN CN CN CN ; ; ; N ; N

[0316]

[0316] In any In any aspect or embodiment aspect or embodiment described describedherein, herein,W²Wis 2 selected from the group is selected from the group nvvv MVV AVVV NVV MVV ^^^^ NVV

N N / / -- NN N- \ nvv

N N N consisting of: consisting of: /N~/ ; N ; N; N

N NN NVV

N N=N N MVV

N~ N N //// N H N F N

N N N N N-N N N N N N N A N N NT NTN

N N N N N N ON'N N N N N N ; , *~N/and ; and

[0317]

[0317] In In any aspect or any aspect or embodiment embodiment described theABM herein,the describedherein, ABM comprises comprises a structure a structure

selected from,butbut selected from, notnot limited limited to the to the structures structures shownshown below, below, where a where a dashed dashed line linethe indicates indicates the attachment point of attachment point of aa linker linker moiety moiety or aa ULM: ULM:

162

Oct 2023

Y² y2 R¹ R 1R R² W¹ N N N Y¹ W² 2023248067 10

ABM-a ABM-a

wherein: wherein:

(R)-4 (R23)0-4 (R23)0-3 (R)- ^^^^

MVV

R22 R22 W¹ is W, is - R or or N R ;

each each RR 22 is isindependently independently H or H or -CN; -CN;

each each RR 23 is isindependently independentlyH, H, halo,oror-CF; halo, -CF3; yl, Y¹, y2 are each Y² are independently O0ororS;S; each independently

R 1, R R¹, 2, are R², are each each independently independently HH or or aa methyl group; methyl group;

W² 2is substituted by by 1, 1,22oror3 3Rw 2 and W is aa bond, C 1-6 bond, C- aryl,or aryl, heteroaryl, each or heteroaryl, optionally substituted each optionally R²;; and each each R²² R2isis independently independently H, H,halo, halo, C- alkyl(optionally alkyl C1-6 (optionallysubstituted substituted by by 11 or or more moreF), F), OC- OC1 3alkyl (optionally alkyl (optionally substituted or 1 by 1by substituted more -F). -F). or more 2 to onetoorone

[0318]

[0318] In any of the In any embodiments of the described embodiments herein, described the W² herein, is covalently theisWcovalently coupled coupled or more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as described herein. described herein.

[0319]

[0319] In certain In certain additional additionalembodiments, embodiments, W¹ 1 is selected W is selected from fromthe the group groupconsisting consisting of: of: CI CF 3 F F CI CI CI CF ^^^^ ^^^^ { MVV { CN CN CN& CN CN CN CN ; ; ; N N and ; and

CF 3 ~~~ CF $

CN N N

[0320]

[0320] In In any aspect or any aspect or embodiment describedherein, embodiment described herein,W2W2 is is selectedfrom selected from thegroup the group

/// ~~~

consisting of: consisting of: and and

163

103211 In any

[0321] In any aspect aspect or embodiment or embodiment described described herein, ABM isABM herein, is selected selected from from the the group group

consisting of:. consisting of:

in s S C1 CI S

o o N7 N \ N k NE N NEEN N+ N N N N \ NN

Fp F N N FF N -\ N S F F SS F F FF F o 0 ~ 00 O o o NC NC N NC N N N 0ON /' 2 N N N ON N F 3C SIQ0 F FC S 0 FC FC S S F FC 0 F 3

0 0 o o 0 O NC- / \ N NC NC /\ NN N NC N NC N N N N S S FS F S inP CI S m 0 00 o o o N0 NC- /\ _N/`+ NC- N N+ NC N NC- NC N N Q F3 0- /N N' N eN F NC N N N FC S MeO MeO S S A S _\ F3C FC SA S O Q0 0F F 0 o o o NC~N N/\N1LNC~N/` NC N NC N N NC N N N P ci sI MeO Sn1 FCI3 C S MeO S FC S 3~ F F0 N N 0 F F o a 0 N~ 0 O O O N o

NC/\NNC NC N N NC NC \ N N NC N N N F 3C SF S S -NCN 3Ce FC S o FC FC S o 0 o 0 o 0 0C- o NC N+ N Nh' NNC-+NC~ -N NN NCFC N N NC N S N F 3C S IF 3C s S I3 FC S FC S FC F N

164

O,, 0 NH' N NH NH N CI o N c 1aNAwho N;0c CI o N IZ N H N H N

O,, NN NH N CI 0, NH O N CI 6, N NA O NHH we N ~ IZ IZ N H NN H CI 0 NN O,

111 NA3 05NA NH H A H N CI NH o N CF o nn IZ

H 0 O,, O,

NH N NH NH NH N CI N N CI O CF O o who

N A o o NH NH N N N N N F S N F F F 0 F S F F H N

NN

N- NN O N H; 0 N N ~~~~

N F N F F FF S K NH HN N NNN

CI CI 0 0

165

H

N NX NI 2023248067 10 Oct

N o 00 N N N N HN N NNI CI

Ci ~ 0 N CI

N 00 you

O HN////, N N NN N N

o S CI cl ~ 00

N

you N N 1J 0 N NN N IZ NH HN N 00 o CIl CI 0 O N

166

N ZI H 'N~ ZI H N N ~ ZI H N ~ HN N HN\N "N ~H N "." NN ZI H N N 0 0 "N -0 0(0 0 O 0 0ec Wy 0 O 0 O CI CI CI K- CI CI C CI C CI N N IIN N N N NN N N

NN N\ N N N ZI N H H HZI H IZ H ,0 "NH "r ZI H HN HN N 11 F 0 00 N 0 0 0 OFF O O O O

CI -l C I; Cl CI c CI C N N N N NN N N N H H/ ZI H N ZI N N O, j, .,",NfNT N\ H N N-- N N N 0"'00 N *."',Ny N N- \ O O 0"" 0 0" O O O CI C CI CI CI N N N N 11III N N N N N

167

Oct 2023 N IZ N ZI ZI H NNZI H H H HN N N N N N N N N

H N Ni NON O CI CI CI 2023248067 10

C C N N N N N N N HN N N N N C N CI ZI ZI N H H CI H CI N ZI N N N N N O HN O N F C NN 0' C O 00 OF O F O O

CI CI C CI C C ICI N NN N N N N N

NNN N / H ZI H NN N / N N 0 N HZI N N

' 01 0"(' H O 0 .L N N N Y- N NN\

[02]In nIseto emodmntdecibe heenIh B N oisstetrcu: I~.0O 0 O o c CI CI

N N N CI CI CI CI C1 N CI

N N N 103221 In any

[0322] In any aspect aspect or embodiment or embodiment described described herein, herein, thecomprises the ABM ABM comprises the structure: the structure:

(R°)-

Y 33 Y Y4Y W2 W² Y W1 w¹

A BM-b ABM-b

wherein: wherein:

W¹ 1isisaryl, W aryl, ororheteroaryl, heteroaryl,each independently each substituted by 1 or more H, halo, hydroxyl, independently substituted by 1 or more H, halo, hydroxyl, nitro, CN, nitro, CN, C=CH, C=CH, C-C1-6 alkyl alkyl (linear,branched, (linear, branched,optionally optionallysubstituted substitutedbyby1 1orormore morehalo, halo, C1.6 alkoxyl),C-C1.6 C- alkoxyl), alkoxyl alkoxyl (linear,branched, (linear, branched,optionally optionallysubstituted substitutedbyby1 1orormore morehalo), halo), C2.6 alkenyl, C2-6 C2-6 alkenyl, alkynyl,or or C2-6alkynyl, CF;CF ; 3

Y³, Y,4 Y Y3 , Y , Yareareeach eachindependently independently a bond, a bond, O, NRY², 2 0, NRYCRY¹RY², C=0,2 , C=S, , CR RY C=0, SO, C=S, SO, SO, SO2 ,

heteroaryl, ororaryl; heteroaryl, aryl;

168

Oct 2023 Q is aa 44 membered Q is alicyclic ring membered alicyclic ring with with 0-2 0-2 heteroatoms, heteroatoms, optionally optionally substituted with 0-6 substituted with 0-6 R°, RQ, each RQ is each R° is independently independently H,H,C-C1alkyl -6alkyl (linear,branched, (linear, branched,optionally optionallysubstituted substitutedbyby1 1oror more halo, C more halo, C1-6 -6 1 alkoxyl), or 2 RQ groups taken together with the atom they are attached to, alkoxyl), or 2 R° groups taken together with the atom they are attached to,

form form aa 3-8 3-8 membered membered ringsystem ring system containing containing 0-20-2 heteroatoms); heteroatoms); 2023248067 10

R 1, RR2 R¹¹, are each ² are each independently independentlyH,H,C-C 1alkyl -6alkyl (linear,branched, (linear, branched,optionally optionallysubstituted substituted by by 11 or or more halo, more halo, C- CI-6 alkoxyl); alkoxyl);

W² 2isisa abond, W bond,C- Calkyl, C1-6 heteroalkyl, O, C- alicyclic, heterocyclic, aryl, biheterocyclic, 1 -6 alkyl, C1 -6heteroalkyl, 0, C1-6 alicyclic, heterocyclic, aryl, biheterocyclic,

biaryl, or biheteroaryl, or heteroaryl, each optionally substituted by 1, 2 or 3 Rw 2 ; and biaryl, or biheteroaryl, or heteroaryl, each optionally substituted by 1, 2 or 3 R²; and

each R2 each R² independentlyH,H,halo, isisindependently halo,C-C1-6 alkyl alkyl (linear,branched, (linear, branched,optionally optionallysubstituted substitutedby by11oror more F), more F), C- C 1 -6heteroalkyl(linear, heteroalkyl (linear, branched, branched, optionally substituted), -ORw optionally substituted), 2 OC-alkyl -ORW²AA OC1-3alkyl

(optionally substitutedby by (optionally substituted 1 more 1 or or more -F), -F), cycloalkyl, C 3 -6 C- cycloalkyl, C4-6 cycloheteroalkyl C4-6 cycloheteroalkyl (optionally (optionally

substituted), C1-6 substituted), C- alkyl (optionally substituted), alicyclic (optionally substituted), alkyl (optionally substituted), C1-6 alicyclic C1-6 (optionally substituted),

heterocyclic (optionally substituted), aryl (optionally substituted), heteroaryl (optionally heterocyclic (optionally substituted), aryl (optionally substituted), heteroaryl (optionally

substituted), bicyclic substituted), bicyclichereoaryl hereoaryl(optionally (optionallysubstituted), bicyclic substituted), aryl,aryl, bicyclic OH,OH, NHNH, , NR 'R 2 NRY¹R², 2

or or CN; and CN; and

RW 2 Aisis H, R²²A H, C- C 1-6 alkyl(linear, alkyl (linear, branched), branched), or or C- C1-6heteroalkyl heteroalkyl(linear, (linear, branched), branched), each each optionally optionally substituted by a cycloalkyl, cycloheteroalkyl, aryl, heterocyclic, heteroaryl, halo, or OCi1 substituted by a cycloalkyl, cycloheteroalkyl, aryl, heterocyclic, heteroaryl, halo, or OC1-

3alkyl. alkyl.

[0323] In any

[0323] In any aspect aspect or embodiment or embodiment described described herein, herein, the description the description provides provides an androgen an androgen

receptor bindingcompound receptor comprising bindingcompound comprising a structure a structure of:of:

(R°)- 2. Y5 W W² Y³ Y Y w¹ Y ABM-e ABM-e

wherein: wherein:

W¹ 1 W isisaryl, heteroaryl,, ,bicyclic, aryl, heteroaryl, bicyclic,ororbiheterocyclic, biheterocyclic, eacheach independently independently substituted substituted by 1 or by 1 or

more H,H,halo, more halo, hydroxyl, hydroxyl, nitro, nitro, CN, C-CH,C1-6 CN, C=CH, C1 -6alkyl(linear, alkyl (linear, branched, branched, optionally optionally substituted by1 1orormore substituted by more halo, halo, C1-6 alkoxyl), C- alkoxyl), C1-6 alkoxyl C1-6 alkoxyl (linear, (linear, branched,branched, optionally optionally

substituted by1 1orormore substituted by more halo), halo), C2-6C2-6 alkenyl, alkenyl, C2-6 C2-6 alkynyl, alkynyl, or CF;or CF ; 3

Y¹, Y²2 are Y', Y are each NR 1O, independently NRY¹, each independently , 0,ororS;S;

169

Oct 2023 Y³, Y,4 Y Y 3, Y , Yareareeach eachindependently independently a bond, a bond, O, NRY², 2 0, NRYCRY¹RY², C=O, 2C=S, , CR RY , C=0,SO,C=S, SO,SO, SO2, heteroaryl, ororaryl; heteroaryl, aryl; Q is aa 3-6 Q is 3-6 membered alicyclic or membered alicyclic or aromatic aromatic ring ring with with 0-4 0-4 heteroatoms, heteroatoms,optionally optionally substituted substituted with 0-6 with 0-6 RQ, each R° R°, each RQ,is independentlyH,H,C1-6 is independently alkyl(linear, C1-6alkyl (linear, branched, optionally branched, optionally 2023248067 10

substituted substituted by by 11 or or more more halo, halo, C1-6 alkoxyl), or C- alkoxyl), or 22 R° RQ groups groupstaken takentogether togetherwith withthe the atom atom they they are are attached attached to, to,form form aa 3-8 3-8membered ringsystem membered ring systemcontaining containing0-2 0-2heteroatoms); heteroatoms); R1, R¹, R2, R², Ra, R, R, R 1,R Ry2 R, R¹¹, areeach ² are eachindependently independently H, H, C1-6 alkyl C- alkyl (linear, (linear, branched, branched, optionally optionally

substitutedbyby1 1orormore substituted more halo, halo, C1-6C1-6 alkoxyl), alkoxyl), orR²R1together or R¹, , R2 together with with the atomthe atom they are they are attached attached to, to, form form aa 3-8 3-8 membered ringsystem membered ring systemcontaining containing0-20-2heteroatoms); heteroatoms); W² 2isisa abond, W bond,C- Calkyl, C-6 heteroalkyl, O, C- alicyclic, heterocyclic, aryl, biheterocyclic, 1 -6 alkyl, C1 -6heteroalkyl, 0, C1-6 alicyclic, heterocyclic, aryl, biheterocyclic,

biaryl, or biaryl, or biheteroaryl, biheteroaryl,ororheteroaryl, heteroaryl, each each optionally optionally substituted substituted by 1, by 2 or1,3 2R²; or 3 Rw 2 each RW2 each R²² is isindependently independently H, halo, H, halo, C- alkyl alkyl (linear, C1-6(linear, branched, branched, optionallyoptionally substitutedsubstituted by 1 or by 1 or 2 more F), more F), C- heteroalkyl(linear, C1-6heteroalkyl (linear, branched, branched, optionally optionally substituted), substituted), -ORw -OR W²A, OC-alkyl A, OC1-3alkyl

(optionally substituted (optionally substituted by by 1 more 1 or or more -F), -F), C 3 -6 cycloalkyl, C3-6 cycloalkyl, C4-6 cycloheteroalkyl, C4-6 cycloheteroalkyl, CI-6 alkyl C1-6 alkyl

(optionally substituted),C-C1-6 (optionally substituted), alicyclic alicyclic (optionally (optionally substituted), substituted), heterocyclic heterocyclic (optionally (optionally

substituted), aryl(optionally substituted), aryl (optionallysubstituted), substituted), or or heteroaryl heteroaryl (optionally (optionally substituted), substituted), bicyclic bicyclic

hereoaryl ororaryl, OH,OH, NHNH, 1 Rv 2, CN; hereoaryl aryl, 2, NR NRY¹R², CN; and and

R²²A2 A RW isisH,H, C1-6 alkyl C 1-6 (linear, alkyl (linear,branched), or C-orheteroalkyl branched), (linear,(linear, C1-6 heteroalkyl branched), each optionally branched), each optionally substitutedbybya acycloalkyl, substituted cycloalkyl, cycloheteroalkyl, cycloheteroalkyl, aryl, aryl, heterocyclic, heterocyclic, heteroaryl, heteroaryl, halo, orhalo, OC1- or OC1. 3alkyl. alkyl.

[0324] In any

[0324] In any aspect aspect or embodiment or embodiment described described herein, herein, an androgen an androgen receptor receptor binding binding moiety moiety

has aa structure has structureof: of: (R°)- 2. Y5 W W² Y³ Y YY

w¹ Y ABM-e ABM-e

wherein: wherein:

(R23)0o4 (R)-4 (R23)0-3 (R)- ^^^^

^^^^

R R22 W, is W¹ is - R or or N R ;

each each RR 2is 2 is independently independently H or H or -CN; -CN;

170

Oct 2023 each each RR 2is 3 isindependently independentlyH, H, halo,oror-CF; halo, -CF3; Y³ 3 is Y is aa bond bondororO;0; Q is aa 44 member Q is ring, optionally member ring, optionally substituted substituted with with 0-4 0-4 RQ, Rº, each each RQ is independently R° is independently HHor or methyl; methyl; 2023248067 10

Y4 is aa bond Y4 is or NH; bond or NH; Y5 is aa bond, Y5 is bond, a a C=O, or aa C=S; C=O, or C=S;and and 2 independently a bond, C1-6 aryl or heteroaryl, each optionally substituted by 1, 2 each Wis each W² is independently a bond, C1-6 aryl or heteroaryl, each optionally substituted by 1, 2 or R²,2 ,each or 33 RW eachR²² Rw2isisindependently independentlyH,H,halo, halo,a a6 6member member alicyclicring alicyclic ringwith with1 or 1 or2 2 heteroatoms ororaa 55 member heteroatoms member aromatic aromatic ring ring with with 1 or2 2oror3 3heteroatoms. 1 or heteroatoms.

[0325]

[0325] In any In any aspect or embodiment aspect or embodiment described describedherein, herein,W²Wis 2 selected from the group is selected from the group nvvr MVV nvvv ^^^^ /VVV nvv NVV NVV

N N N N consisting of: consisting of: N N N N , , ,

N NN N NWV

MVV 0 N=N N N N N nnn

H , , ;; and and F F

2

[0326]

[0326] In any In any aspect or embodiment aspect or embodiment described describedherein, theW²Wis herein,the iscovalently covalentlycoupled oneoror coupledtotoone more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as described herein. described herein.

[0327]

[0327] In any aspect In any or embodiment aspect described or embodiment herein, described W¹ is W herein, selected from the is selected fromgroup the group CI CF 3 F F CI CI CI CF ~~~ $ MVV nnv ^^^^

CN. CNCN C CN CN CN CN consisting of: consisting of: ; ; ; ; ; NN N ;

Me We CF Me OMe ~~~ CF3 s S CN CN S CN CN and CN ; and N

[0328]

[0328] In In any aspect or any aspect or embodiment embodiment described herein,ananandrogen describedherein, androgen binding binding moiety hashas moiety a a

structure of: structure of:

171

Oct 2023

(R²)- Y³ W² (Y³)-5

w¹

2023248067 10 ABM-d ABM-d

wherein: wherein:

W¹ 1is W is aryl, aryl, independently substituted by independently substituted by 11 or or more more halo, halo, CN; CN;

Y3 Y³ are areeach independently each a bond, independently a NRY 2 bond, 1 RY 2C=O; NRY², , CR , C=O; Q is aa 55 membered Q is aromaticring membered aromatic ringwith with1 1oror2 2heteroatoms; heteroatoms; R¹¹,, Ry2are R each independently R ² are each independently H,H,C-C1-6alkyl (linear,branched); alkyl (linear, branched); W² 2isisa abond, W bond,aryl, aryl,ororheteroaryl, heteroaryl, each each optionally optionally by 1, 2by substituted substituted or 1, 3 2 R²; 3 Rw 2; and orand each RW2isis independently each R²² independently H,H,halo, halo,C1-6alkyl (optionallysubstituted C- alkyl (optionally substituted by by 11 or or more moreF), F), OC1- OCi 3alkyl (optionally alkyl (optionally substituted or 1 by 1by substituted more -F). -F). or more 2

[0329]

[0329] In any aspect In any or embodiment or embodiment aspect described described herein, herein, is W the W²the is covalently covalently coupled to onetoorone coupled or more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as described herein. described herein.

(R23)0o4 (R)-4 ^^^^^

[0330] In any

[0330] In any aspect aspect or embodiment or embodiment described described herein, W¹ is W 1 is herein, - R ;

wherein each wherein eachRRis 22 is independently independently halo halo or or CN;CN; and and each 3 is each RR 2is independentlyH or independently H or halo. halo.

[0331] In any

[0331] In any aspect aspect or embodiment or embodiment described described herein, W¹ is W1 herein, selected from the is selected fromgroup the group consisting of: consisting of:

CI CI

FF C1 CI F F C1 CI ~~~ $ \CN CN ~~~ MVV ^^^^

$ CN CN CN CN ON; CN ;and ; ; ; and F F

23 NN N

[0332] In any

[0332] In any aspect aspect or embodiment or embodiment described described herein, herein, Q is Q is

172

^^^^ N-NH N-NH

[0333]

[0333] In any aspect In any or embodiment aspect described or embodiment herein, described W² is W 2 herein, is { NVV

(R) ' (R)N NH NH nnn

[0334]

[0334] In any aspect In any or embodiment aspect or embodiment described described herein, herein, is 3 )o-s (Y³). (Y is 0 2023248067

In any

[0335] In any

[0335] aspect aspect or embodiment or embodiment described described herein, herein, the ABM ABM comprises thecomprises a structure a structure

selected from,butbut selected from, notnot limited limited to the to the structures structures shownshown below, below, where a where a dashed dashed line linethe indicates indicates the attachment point of attachment point of aa linker linker moiety moiety or or aa ULM, suchasasaaCLM: ULM, such CLM:

R1 R2 Y² R1 y2 R¹ R² R R w¹ N R2 N Y N Y³ Y¹ W² W² Y ABM-a ABM-a W R2 R R R W N N

N N N--NH NH NNHNH ///////// NH

W2 0 O W wherein: wherein:

(R23)0-4 (R)-4 (R23)0-3 (R)- ^^^^

^^^^

R22 R22 W, is W¹ is - R or or N R ;

each each RR 2is 2 is independently independently H or H or -CN; -CN;

each each RR 2is 3 isindependently independentlyH, H, halo,oror-CF; halo, -CF3; Y', Y¹, Y2 are each Y² are independently O0ororS;S; each independently

Y³, Y,4 ,YYareare Y3 , Y each eachindependently a bond, a bond, independently O, 0, NRY², 2 NR CRY¹RY², 2 C=S, so, or SO;; , CRRYC=O,, C=0, C=S, SO, or SO 2 ;; R 1, R R¹, 2, are R², are each each independently independently HH or or aa methyl group; methyl group;

W² 2isisa abond, W bond,C1-6 C1 aryl, or or -6 aryl, heteroaryl, eacheach heteroaryl, optionally substituted optionally by 1, 2by substituted or 1, 3 R²; 3 Rw 2 ; and 2 orand

173

Oct 2023 each each R²² R2isis independently independently H, H,halo, halo, C- Ci-6 alkyl(optionally alkyl (optionallysubstituted substituted by by 11 or or more moreF), F), C3-6 C 3 -6 cycloalkyl, cycloalkyl, C4-6 C4-6cycloheteroalkyl, cycloheteroalkyl,OCi-3alkyl (optionally substituted OC-alkyl (optionally substituted by by 11 or or more more-F). -F). 2

[0336]

[0336] In any aspect In any or embodiment aspect described or embodiment herein, described the W²the herein, is W covalently coupled is covalently to onetoorone coupled or more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as 2023248067 10

described herein. described herein.

[0337]

[0337] In any In any aspect or embodiment aspect or embodiment described describedherein, 1 isselected herein,W¹Wis selectedfrom fromthe thegroup group consisting of: consisting of:

F CI1CI CI CF 3 CF 3 CF F CF $ $ \/ CN MVV ^^^^

CN MVV

CN. 1 CN. CN CN CN CN CN CN ; ; ; N ; N ;

N N CN and and N

[0338]

[0338] In In any aspect or any aspect or embodiment describedherein, embodiment described herein,W2W2 is is selectedfrom selected from thegroup the group

nvv ////

N N N N consisting of consisting of: , N ,

N N N

, and and

[0339]

[0339] In In any aspect or any aspect or embodiment embodiment described herein,the describedherein, ABM theABM comprises comprises a structure a structure shown shown

below, where below, wherea adashed dashedline lineindicates indicates the the attachment attachment point point of of aa linker linker moiety or a ULM moiety or ULM orora aCLM: CLM:

R¹ R² R2 R² R¹ R1 R2 (Y W² R¹ (Y)- R¹

your 3 Y R¹ R² R R2 R1 R2 R R² R R¹

W¹

wherein: wherein:

174

Oct 2023

(R23)0-4 (R)-4 (R)- (R23)0-3 ^^^^

MVV

-R22 W R22 r W W¹ is is i R eor or N R ;

2023248067 10 each each R 2 is R 2is independently independently Hor H or -CN; -CN;

each is isindependently each RR 23 independentlyH, H, halo,oror-CF; halo, -CF3; Y³ 3is Y is aa bond bondororO;0; Y 4isis aa bond Y bondoror NH; NH; Y 5isis aa bond, Y bond,C=O, C1 heteroaryl, C=O, C-C -C6 heteroaryl, orCaryl; or C-C 1 -C6 aryl;

R 1, R R¹, 2, are R², are each each independently H, or independently H, orCI-C6alkyl (linearororbranched, C-C alkyl (linear branched,optionally optionallysubstituted substituted by 11 or by or more halo, orC more halo, alkoxyl); or C- 6alkoxyl);

W² 2 is W is aa bond,C 1 .aryl, bond, C- 6 aryl,C- 6 heteroaryl,C C- heteroaryl, .6 alicyclic,ororC C- 1alicyclic, -6 heterocyclic, C- 1heterocyclic, each each optionally optionally

substituted substituted by by 1-10 R²; 2;and 1-10 Rw and each each R² R2 isisindependently independentlyH,H,ororhalo; halo;and and

[0340]

[0340] representsa abond ~~ represents bond that that maymay be stereospecific (S))or ((R) or((R) be stereospecific or(S)) or non-stereospecific. non-stereospecific.

2 to onetoorone

[0341]

[0341] In any of the of the In any embodiments embodiments described described herein, herein, the W²theisWcovalently is covalently coupled coupled or more ULM more ULMor or CLMCLM groups, groups, or a or a linker linker to which to which is attached is attached one one or more or more ULM ULM or CLM or CLMasgroups groups as described herein. described herein.

[0342] In certain

[0342] In certain additional additional embodiments, embodiments, 1 is selected W¹ isWselected from from the group the group consisting consisting of: of:

CF 3 F F CI CC1CI CF 3 CF CF $ ~~~

CN MVV \ CN //// NVV CN CN CN CN CN CN CN CN ; ; ; N ; N

[0343]

[0343] In certain In certain additional additionalembodiments, W 2isis selected embodiments, W² selected from fromthe the group groupconsisting consisting of: of: nvvv NVV nvv nnn NVV MVV ^^^^ MVV ^^^^

-NN N N -N N N N N ; ; N ; N ; ;

^^^^

N N=N N/N N N N nnn

N H H 5 N - ~ .F F ; NN ;

175

N N N NN N N NN N NN N N NN N N N N ; N ; ; , N N N N

2023248067 N N N ; ;and ; and N

[0344] In certain

[0344] In certain embodiments, embodiments, the androgen the androgen receptor receptor binding binding compound compound of ABM is ABM is ofselected selected from thegroup from the group consisting consisting of: of:

trans-2-Chloro-4-[3-amino-2,2,4,4-tetramethylcyclobutoxy]benzonitrile; trans-2-Chloro-4-[3-amino-2,2,4,4-tetramethylcyclobutoxy]benzonitrile;

cis-2-Chloro-4-[3-amino-2,2,4,4-tetramethylcyclobutoxy]benzonitrile; cis-2-Chloro-4-[3-amino-2,2,4,4-tetramethylcyclobutoxybenzonitrile;

trans trans 6-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyridazine-3 6-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyridazine-3-

carboxamide; carboxamide;

trans tert-Butyl trans N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]carbamate; tert-ButylN-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]carbamate;

trans trans 4-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide; 4-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide;

trans trans 5-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyrazine-2 5-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetraethylcyclobutyl]pyrazine-2-

carboxamide; carboxamide;

trans 2-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyrimidine-5 trans 2-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyrimidine-5-

carboxamide; carboxamide;

4-Methoxy-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide; 4-Methoxy-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide;

trans trans 1-(2-Hydroxyethyl)-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]-1H 1-(2-Hydroxyethyl)-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyI]-1H

pyrazole-4-carboxamide; pyrazole-4-carboxamide;

trans 6-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyridine-3 trans 6-Amino-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]pyridine-3-

carboxamide; carboxamide;

trans 4-[(5-Hydroxypentyl)amino]-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4 trans4-[(5-Hydroxypentyl)amino]-N-[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl]benzamide; and tetramethylcyclobutyl]benzamide; and

trans tert-Butyl trans tert-Butyl2-({5-[(4-{[3-(3-chloro-4-cyanophenoxy)-2,2,4,4 2-({5-[(4-{[3-(3-chloro-4-cyanophenoxy)-2,2,4,4

tetramethylcyclobutyl]carbamoyl}phenyl)aminopentyl}oxy)acetate; tetramethylcyclobutyl]carbamoyl}phenyl)aminopentyl}oxy)acetate;and and N-((1r,3r)-3-(4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-methylbenzamide. N-(1r,3r)-3-(4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-methylbenzamide.

XIII.

[0345] XIII.

[0345] Compounds Compounds Targeting Targeting Estrogen Estrogen Receptor (ER) (ER) Receptor ICI-182780 ICI-182780

176

Oct 2023

[0346] 1. Estrogen

[0346] 1. Estrogen Receptor Receptor Ligand Ligand

OH OH H H 2023248067 10

HH H H H ZI H N R

[03471

[0347] HO Ho R

[0348] (Derivatized

[0348] (Derivatizedwhere where "R""R" designatesa site designates a site for for linker linker group group LL or -(L-CLM)group or -(L-CLM) group attachment). attachment).

[0349] In In

[0349] anyany embodiment embodiment or aspect or aspect describedherein, described herein, the the PTM maybeberepresented PTM may represented by by the the Formula PTM-I: Formula PTM-I:

XPTM1- XPTM1 .- RPTM3 PTM3- XPTM2

XPTM XpTM

RPTM2 RPTM2 RPTM1 RPTM1 S

PTM-I PTM-I wherein: wherein:

XPTM is OO or XPTM is C=O; or C=O; each each of of XPTM1 andXPTM2 XPTMI and XPTM2is isindependentlyselected independently selectedfrom fromN N or or CH; CH;

RPTM1 is independently RPTM1 is independentlyselected selectedfrom from OH,OH, O(CO)RPTM, O(CO)RPTM, 0-lower O-lower alkyl, wherein alkyl, wherein RPTM is RPTM an is an alkyl or aryl alkyl or aryl group groupin inthetheester; ester; at at least least one RPTM2, each one RPTM2, independentlyselected each independently selectedfrom fromH, H, OH,OH, halogen, halogen, CN, CN, CF 3 , S02-alkyl, CF, SO-alkyl,

0-lower alkyl; O-lower alkyl;

at at least leastone RPTM3, each oneRPTM3, each independently selected from independently selected H, halogen; from H, halogen; and and the dashed the dashedline line indicates indicates the the site site of of attachment attachmentofofatatleast least one onelinker, linker, CLM, CLM, CLM', CLM', PTM, PTM, PTM', PTM', or a or a combinationthereof. combination thereof.

[0350] In In

[0350] anyany embodiment embodiment or aspect or aspect describedherein, described herein, the the PTM maybeberepresented PTM may represented by by the the Formula PTM-I: Formula PTM-I:

177

Oct 2023

XPTM1- XPTM1 RPTM3-- RPTM3 XPTM2 XPTM2

XPTM XpTM 2023248067 10

RPTM2_ RPTM2 RPTM2RPTM1 RPTM1 S

PTM-I PTM-I wherein: wherein:

XPTM is O0or XPTM is C=O; or C=O; each each of XPTM1 andXPTM2 XPTM1 and XPTM2is isindependentlyselected independently selectedfrom fromN N or or CH; CH;

RPTM1 is independently RPTM1 is independentlyselected selectedfrom from OH,OH, O(CO)RPTM, O(CO)RPTM, 0-lower O-lower alkyl, wherein alkyl, wherein RPTM is RPTM an is an alkyl or aryl alkyl or aryl group groupin inthetheester; ester; each is independently each RPTM2 is independentlyselected selectedfrom H,H,OH, from OH,halogen, halogen,CN, CN,CF 3 , S02-alkyl, CF, SO-alkyl, 0-lower O-lower

alkyl; alkyl;

each RPTM3 isisindependently each RPTM3 independently selected selected from fromH,H,halogen; halogen; the the PTM-I comprises PTM-I comprises as as leastone least oneRPTM2, at at RPTM2, leastone least oneRPTM3, RPTM3,or ora acombination combination thereof thereof on on thethe

respectiverings; respective rings;and and the dashed the dashedline line indicates indicates the the site site of of attachment attachmentofofatatleast least one onelinker, linker, CLM, CLM, CLM', CLM', PTM, PTM, PTM', PTM', or a or a combinationthereof. combination thereof.

[0351] In

[0351] In any any embodiment embodiment or or aspect aspect described described herein,PTM-I herein, PTM-I has has at least at least oneone of:of:two two RPTM2, RPTM2, twotwo

RPTM3, or aa combination RPTM3, or thereof. combination thereof.

[0352] In

[0352] In any any embodiment or aspect embodiment or aspect described described herein, herein,the thePTM maybeberepresented PTM may represented by by the the Formula PTM-II: Formula PTM-II:

RPTM5 XPTM1 RPTM5 XPT \I\1' RPTM3 XPTM2

RPTM4 XPTM XpTM

RPTM2 RPTM2 R RPTM1 S RPTM1 S

PTM-II PTM-II wherein: wherein:

178 is O0or 2023248067 10 Oct 2023

XPTM XPTM is C=O; or C=O;

eachof each of XPTM1 andXPTM2 XPTM1 and XPTM2 is isindependentlyselected independently fromN N selectedfrom or or CH; CH;

RPTM1 is independently RPTM1 is independentlyselected selectedfrom from OH,OH, O(CO)RPTM, O(CO)RPTM, O-lower O-lower alkyl, wherein alkyl, wherein RPTM is RPTMis an an alkyl or aryl alkyl or aryl group groupin inthetheester; ester; andRPTM4 RPTM2 and RPTM2 RPTM4 areindependently are independently selected selected from from H, halogen, H, OH, OH, halogen, CN, CN, CF, CF 3 , S02-alkyl, SO2-alkyl, O- 0 lower alkyl; lower alkyl;

andRPTM5 RPTM3 and RPTM3 areindependently RPTM5are independentlyselected selectedfrom from H,H, halogen; halogen; andand

the dashed the dashedline line indicates indicates the the site site of of attachment attachmentofofatatleast least one onelinker, linker, CLM, CLM, CLM', CLM', PTM, PTM, PTM', PTM', or a or a combinationthereof. combination thereof.

[0353] In aspect

[0353] In embodiment aspect ororembodiment described described herein, herein, O(CO)RPTM O(CO)RPTM functions functions as a of as a prodrug prodrug the of the corresponding phenolininFormula corresponding phenol Formula PTM-I PTM-I or PTM-II. or PTM-II.

any embodiment In any

[0354] In

[0354] embodiment or or aspect aspect described described herein, O-lower herein,thetheO-lower alkyl of of alkyl PTM-I PTM-I or PTM-II or PTM-II an an alkyl alkyl chain chain with with carbon number1 1toto3.3. carbon number

[0355]

[0355] In In aspect embodiment aspect ororembodiment described described herein, herein, the present the present disclosure disclosure provides provides a compound a compound or or PTMof of PTM Formula(IPTM): Formula (IPTM):

RPTM4 XPTM XpTM zPTM XpTM | RPTM1 || XpTM XPT

RPTM3 RPTM3 PM XpTM XPTM

RPTM2 RPTM2

Formula(IPMT) Formula (IPMT)

wherein: wherein:

eachXPTM each XPTM isisindependently independently CH,CH, N; N;

3~~ indicates indicates the the site site of of attachment attachment of at least of at least one one linker, linker, CLM, CLM',PTM, CLM, CLM', PTM, PTM',PTM', or a or a

combination thereof; combination thereof;

each RPTMI isisindependently each RPTMI independentlyOH,OH, halogen, halogen, O(CO)RPTM,where O(CO)RPTM, or alkyl RPTMis where RPTM is alkyl or cycloalkyl cycloalkyl group withgroup with

11 to to 66 carbons or aryl carbons or aryl groups, substitution can groups, substitution can be be mono-, mono-,di-di-orortri-substituted; tri-substituted; each RPTM2 isis independently each RPTM2 independently H,H,halogen, halogen,CN,CN, CF, CF 3, alkoxy, alkoxy, substitution substitution can can be mono- be mono- or di- or di

substitution; and substitution; and

179

Oct 2023 each each RPTM3 RPTM3 is independently is independentlyH,H, halogen, halogen, substitution substitution cancan be be mono- mono- or di-substitution. or di-substitution.

[0356]

[0356] In In any aspect or any aspect or embodiment embodiment described described herein, herein, the the PTM PTM is represented by theby is represented Formula (): the Formula (IIPTM):

s

2023248067 10 RPTM1 ~

T RPTM3 RPTM3 XpTM

RPTM2 RPTM2

Formula (IIPMT) Formula () wherein: wherein:

XPTM is XPTM CH,N;N; is CH,

rrr indicates indicates the the site siteof ofattachment of at attachment of at least leastone one linker, linker,CLM, CLM', CLM, CLM', PTM, PTM',PTM', PTM, ULM, an ILM, an ILM, ULM, aa VLM, MLM, VLM, MLM, a ULM', a ULM', a ILM', a ILM', a VLM', a VLM', a MLM',a or MLM', or a combination a combination thereof;thereof;

each RPTMI isis independently each RPTM1 independentlyOH,OH, halogen halogen (e.g., (e.g., F);F);

each RPTM2isisindependently each RPTM2 independently H, halogen H, halogen (e.g., (e.g., F), substitution F), CF, CF3 , substitution can becan be or mono- mono- or di-substitution; di-substitution;

and and

each each RPTM3 RPTM3 is independently is independentlyhalogen halogen (e.g. (e.g. F),F), substitutioncancan substitution bebe mono- mono- or di-substitution. or di-substitution.

[0357]

[0357] In certain In embodiments, at at certain embodiments, leastone least of: oneof:

XPTM of XPTM ofFormula Formula(IIPTM) () isis CH; CH;

RPTMI RPTM1 of of Formula Formula (IIPTM) is OH; () is OH;

RPTM2 of Formula RPTM2 of Formula (IIPTM)isH;H; (IIPTM)is

each RPTM3 RPTM3 of of Formula Formula (IIPTM) is independentlyH H () is independently F;F;or oror or

a combination thereof. combination thereof.

[0358] XIV.

[0358] XIV.Compounds Compounds Targeting Targeting ThyroidHormone Thyroid Hormone Receptor(TR) Receptor (TR)

[0359]

[0359] 1. Thyroid 1. Hormone Thyroid Hormone Receptor Receptor Ligand Ligand (derivatized) (derivatized)

180

Oct 2023

MOMO OH OH MOMO O 0 O 2023248067 10

H ZI H N' R N R

[03601

[0360] O

[0361] (Derivatized

[0361] (Derivatizedwhere where"R""R" designatesa site designates a site for for linker linker group group LL or -(L-CLM)group or -(L-CLM) group attachment attachment and andMOMO indicates aa methoxymethoxy MOMO indicates group). methoxymethoxy group).

XV.

[0362] XV.

[0362] Compounds Compounds targeting HIV targetingHIV Protease Protease

[0363] 1. Inhibitor

[0363] 1. Inhibitor of HIV of HIV Protease Protease (derivatized) (derivatized)

Ph~ Ph O HO ZI H O R R'N N N N N ZI O N N N ~KOHH

[0364]

[0364] O Ph Ph OH

[0365] (Derivatized

[0365] (Derivatizedwhere where "R""R" designates designates a sitefor a site forlinker linker group group L Lor-(L-CLM) or-(L-CLM) group group

attachment). See, J attachment). See, J. Med. Med. Chem. 2010,53, Chem. 2010, 53,521-538. 521-538.

[0366] 2. 2.

[0366] Inhibitor Protease HIVProtease InhibitorofofHIV

OH ZI OH 0N HH N N O S O Ph O R'N R Ph O O N

[03671

[0367] H

[0368] (Derivatized

[0368] (Derivatized wherewhere "R" designates "R" designates a potential a potential sitelinker site for for linker group group L or -(L-CLM) L or -(L-CLM)

group attachment). See, group attachment). See, J. J Med. Chem.2010, Med. Chem. 2010,53,53,521-538. 521-538.

[0369] XVI.

[0369] XVI.Compounds Compounds targetingHIV targeting HIVIntegrase Integrase

[0370] 1. Inhibitor

[0370] 1. Inhibitor of HIV of HIV Integrase Integrase (derivatized) (derivatized)

181

R'O0 R O ,0111

MeO MeO N N OH OH F F 0 0 O O 2023248067

CI

[03711

[0371] CI

[0372] (Derivatized

[0372] (Derivatizedwhere where "R""R" designatesa site designates a sitefor for linker linker group group LL or or -(L-CLM) -(L-CLM)group group attachment). See, J attachment). See, J. Med. Chem.2010, Med. Chem. 2010,53, 53,6466. 6466.

[0373] 2. Inhibitor

[0373] 2. Inhibitor of HIV of HIV Integrase Integrase (derivatized) (derivatized)

OH OH ,0111

MeO N N MeO O R F F 0 0 O O

[0374]

[0374] CI CI

[0375] 3. Inhibitor

[0375] 3. Inhibitor of HIV of HIV integrase integrase Isetntress Isetntress (derivatized) (derivatized)

F F o

NNN OH OH N N HO N HN R NH SNNH O N

[0376]

[0376] 0 o 0 o

[0377] (Derivatized

[0377] (Derivatizedwhere where "R""R" designatesa site designates a sitefor for linker linker group group LL or or -(L-CLM) -(L-CLM)group group attachment). See, J attachment). See, J. Med. Chem.2010, Med. Chem. 2010,53, 53,6466. 6466.

[0378] Compounds XVII.Compounds

[0378] XVII. targeting HCV targetingHCV Protease Protease

[0379] 1. Inhibitors

[0379] 1. Inhibitors of HCV of HCV Protease Protease (derivatized) (derivatized)

182

Oct 2023

NH NH S S N N 2023248067 10

N N 0 On

MeO

/ MeO tBu tBu N N N4H O O C02H COH NH NH NH OO R-o O O

[0380]

[0380] R-O 0 N

[0381] (Derivatized

[0381] (Derivatizedwhere where "R""R" designatesa site designates a sitefor for linker linker group group LL or or -(L-CLM) -(L-CLM)group group attachment). attachment).

[0382] XVIII.

[0382] XVIII. Compounds Compounds targeting targeting Acyl-protein Acyl-protein Thioesterase-1 and -2and Thioesterase-1 -2 and (APT1 (APTi and APT2) APT2)

[0383] 1. Inhibitor

[0383] 1. InhibitorofofAPT1 APTand IandAPT2 APT2 (derivatized) (derivatized)

Me N 2 MeN

s=o S=O R R N-N N-N N.

[0384]

[0384] O

[0385] (Derivatized

[0385] (Derivatizedwhere where "R""R" designatesa site designates a sitefor for linker linker group group LL or or -(L-CLM) -(L-CLM)group group attachment). See, Angew. attachment). See, Angew.Chem. Chem. Int. Int. Ed.Ed. 2011, 2011, 50, 50, 98389838 -9842, -9842, where where L is a L is a linker linker group as group as

otherwise described herein otherwise described herein and andsaid said CLM CLM group group is otherwise is as as otherwise described described herein herein such such that that -(L--(L

CLM) bindsthetheCLM CLM) binds CLM group group to a to a PTMgroup PTMgroup as otherwise as otherwise described described herein.herein.

[0386] Compound VIV.Compound

[0386] VIV. targetingTau targeting Protein TauProtein In any

[0387] In any

[0387] aspect aspect or embodiment or embodiment described described herein, herein, themay the PTM PTM may include include a Tau protein a Tau protein

binding moieties. For binding moieties. Forexample, example,the thePTM PTMmaymay be represented be represented by Formula by Formula I, Formula I, Formula II, Formula II, Formula

183

III, III,Formula Formula IV, Formula V,Formula Formula V, Formula VI,Formula, VI, Formula, VII, VII, Formula, Formula, VIII, VIII, Formula Formula IX, IX, Formula Formula X, X, or or Formula XI: Formula XI:

LPTM LPTM D A B A B C LPTM D I II

2023248067

A B C LPTM LPTM D E A B E LPTM D III III IV IV QA LPTM LPTM B LPTM C -- LPTM D V V

A -- LPTM LPTM I(:XcD B C LPTM LPTM A B LPTM LPTM C LPTM D

VI VI VII VII D -- LPTM - A- LPTM B B LPTM C LPTM E VIII VIII

A B LPTM C --LPTM LPTM A LPTM D x X LXX IX

A LPTM D LPTM - :Ki- B C LPTM LPTM F

XI XI ,

wherein: wherein:

184

A, B, B,C,C,D,D,E,E,andand F are independently selected from anfrom an optionally substitutedsubstituted 5- or 6 2023248067 10 Oct 2023

A, F are independently selected optionally 5- or 6-

membered membered heteroarylring, arylororheteroaryl aryl ring, an an optionally substituted 4- optionally substituted 4- to to7-membered cycloalkyl 7-membered cycloalkyl

or a heterocycloalkyl, or a where heterocycloalkyl, where contact contact between between circles circles indicates indicates ring and ring fusion; fusion; and LPTMis is LPTM selectedfrom selected from a bond, a bond, an alkyl, an alkyl, an alkenyl an alkenyl or an alkynyl, or an alkynyl, optionally optionally interrupted interrupted by one by one or more or morerings rings(i.e., (i.e., cycloalkyl, cycloalkyl,heterocycloalkyl, heterocycloalkyl, aryl aryl or heteroaryl), or heteroaryl), or one or or one moreor more functional functional groups selected from groups selected the groups from the groups -0-, -0-,-S-, -S-, -NRPTM- (whereRRPTM -NR PTM- (where PTM isis selected selected from HHor from alkyl), -N=N-, or alkyl), -S(O)-, -SO-, -N=N-, -S(O)-, -S2-,-C(O)-, -C(O)-,-NHC(O)-, -NHC(O)-, -C(O)NH-, -C(O)NH-, -NHSO 2-, -NHSO2-,

NHC(O)NH-,-NHC(O)O-, NHC(O)NH-, -NHC(O)O-, or or -OC(O)NH-, -OC(O)NH-, wherein wherein the said the said functionalgroup functional groupare are optionally locatedat ateither optionally located eitherendend of the of the linker. linker.

[0388] In any

[0388] In any aspect aspect or embodiment or embodiment described described herein, herein, arylheteroaryl aryl and and heteroaryl rings rings of A,of B,A,C,B,D,C, D, E, and F of and F of PTM areoptionally PTM are optionallysubstituted substituted with with 1-3 1-3 substituents substituents each each independently independentlyselected selected from alkyl, alkenyl, from alkyl, alkenyl, haloalkyl, haloalkyl,halogen, halogen,hydroxyl, hydroxyl, alkoxy, alkoxy, fluoroalkoxy, fluoroalkoxy, amino, alkylamino, amino, alkylamino,

dialkylamino, acylamino, dialkylamino, acylamino,trifluoromethyl, trifluoromethyl, and andcyano, cyano,wherein whereinthe thesaid saidalkyl alkyland andalkenyl alkenylgroups groups are further optionally are further optionallysubstituted. substituted.

[0389] In any

[0389] In any aspect aspect or embodiment or embodiment described described herein, herein, the rings the rings of atof at least least one one of B, of A, A, C, B, F, C, F, or or aa combination thereof is combination thereof is selected selected from from optionally optionally substituted substituted5-5-oror6-membered aryl or 6-membered aryl or heteroarylrings; heteroaryl rings; In any

[0390] In any

[0390] aspect aspect or embodiment or embodiment described described herein, herein, the PTM PTM thehas thehas the chemical chemical structure structure of of FormulaI,I, wherein: Formula wherein: A, BB and A, and CCrings rings are are independently independently5-5- or or 6- 6- membered membered fused fused arylororheteroaryl aryl heteroarylrings; rings; LPTM isisselected LPTM selected from a bond from a or an bond or an alkyl, alkyl, and and

D is D is selected selected from from a a 6-membered aryl,heteroaryl 6-membered aryl, heteroaryl or or heterocycloalkyl, heterocycloalkyl, wherein A, wherein A, B, B, CCand andD Dare areoptionally optionallysubstituted substituted with with alkyl, alkyl, haloalkyl, haloalkyl, halogen, halogen, hydroxyl, hydroxyl,

alkoxy, amino, amino, alkylamino, alkylamino,dialkylamino dialkylaminoororcyano. cyano. In any

[0391] In any

[0391] aspect aspect or embodiment or embodiment described described herein, herein, The The PTM has the has PTM the chemical chemical structure structure

of Formula I, wherein: Formula I, wherein:

A and A andCCare are aa phenyl phenyloror aa 6-membered 6-membered heteroaryl heteroaryl ring; ring;

B is B is aa 5-membered heteroarylring; 5-membered heteroaryl ring; LPTM isis a bond; LPTM and bond; and

D is D is a 6-membered heteroarylorora a6-membered 6-membered heteroaryl 6-membered heterocycloalkyl heterocycloalkyl ring; ring;

185

Oct 2023 wherein each wherein eachA, A, B,B,CCand andD Disisoptionally optionallyindependently independentlysubstituted substitutedwith withalkyl, alkyl, haloalkyl, haloalkyl, halogen, hydroxyl, hydroxyl, alkoxy, alkoxy, amino, amino,dialkylamino dialkylaminoororcyano, cyano,and and wherein wherein a nitrogen a nitrogen atom atom

of any ofofthe of any theA,A,B,B,C and C and D rings D rings is directly is not not directly connected connected to a heteroatom to a heteroatom or to a carbon or to a carbon

atom, towhich atom, to which another another heteroatom heteroatom is directly is directly attached. attached. 2023248067 10

[0392] In any

[0392] In any aspect aspect or embodiment or embodiment described described herein, herein, the PTM PTM thehas thehas the chemical chemical structure structure of of Formula III or Formula III or IV, wherein A, BBand wherein A, are5-5- or andC Care or 6- 6- membered membered fused fused aryl aryl oror heteroarylrings, heteroaryl rings, LPTM isisselected LPTM selected from a bond from a or an bond or an alkyl, alkyl, and and D and EE are D and 5- or 6-membered are 5- fusedaryl 6-membered fused aryloror heteroaryl rings,wherein heteroaryl rings, wherein A, C,B,D C, A, B, andDE and E are optionally are optionally substituted substituted with with alkyl, alkyl, haloalkyl, haloalkyl,

halogen, hydroxyl, halogen, hydroxyl, alkoxy, alkoxy, amino, amino,alkylamino, alkylamino,dialkylamino dialkylamino or or cyano. cyano.

[0393] In any

[0393] In any aspect aspect or embodiment or embodiment described described herein, herein, theisPTM the PTM is represented represented by following by following

chemical structure: chemical structure:

HN R¹ O HN N S R1 O O N0 NH2 NH N ON NH R 1Orij N NHS 2 N O O R O NH

R4 OHO R OH o R6 OH ''OH N\ N NHNH O N S NH N R5 0 N R O N N N N M MeO S N R N N

N NH R N N O

CN CN 0186 R¹ CN N 186 R²

N N 6 N R6 K, - N \R S R R¹ S R R¹

N N N N R² N R² N

NI~ N R6 s N NR6 N R R¹ N R R¹ R³ NNH R³ N N R² R² 2023248067 N N R6 ;,tN R6 N R R¹ N N. R R¹

N N N N N R³ R² R³ R²

QN N N1\/ R R F F 6 31 R

R¹ %R 1 N RR0 NR N N N. RNNR R¹ F,

C1 N N N N N N N R³ R² R³ R² N N R¹ O F. N 0 F N R¹ N NH N o S N N N NN R³ N F ,

wherein: wherein:

R', R²2 and R¹, R R3 are and R³ are independently fromH,H,methyl, selected from independently selected ethyl, 2-fluoroethyl methyl,ethyl, 2-fluoroethyl and 2,2,2 and 2,2,2- trifluoroethyl; trifluoroethyl;

5 andR Rare R and R4 are independently independently selected selected from from H, H, methyl, methyl, ethyl ethyl andand halogen; halogen; and and R'isis 11 to R to 22 substituents substituents independently independently selected selected from H, methyl, from H, methyl, ethyl ethyl and and halogen, halogen, wherein the wherein the PTM PTMis iscoupled coupledtotoa aULM ULMvia via L. L.

[0394] In any

[0394] In any of the of the aspects aspects or embodiments or embodiments described described herein, herein, the is the PTM PTM is covalently covalently

coupled to one coupled to one or or more moreULM ULM (VLM (VLM or CLM) or CLM) groups,groups, or a linker or a linker to which to which is attached is attached one orone or

more ULM more ULM(VLM (VLM or or CLM) CLM) groups groups as describedherein. as described herein. In any

[0395] In any

[0395] aspect aspect or embodiment or embodiment described described herein, herein, PTM isPTM is represented represented by chemical by chemical

structure: structure:

187

N NR8-N N R8 R R Rx R 7 / R R 7 /R & N Rx R R R 6d N N z- N N NN "A"

N N " NN NN 8 R R7 R N Rx N 7 /9 N NN W 6d R /R9 N NN R N -N R R R RI N in N N 1 TR F N iN' R 8 N' N N & N

N N N N N R N N Rx R N R R R R "R" N N 8. BL "A"

N N- R8N N 7 / R RN9 7

NN RB N' - N '

N 1 N8 & RNN 8 NR

R X-Z R N R N N N Rx N 2-2 & N R N R10 N R10 N R10 R R8 R8 R N N N N N N N N N

N Qlo 1 N RlQ 7 N No~ R R N R & N R11 N N N R8 N N N 8 N N N R NRR R7 N lo R- /-N,-N4Rl R NR Rx Rx N N R N N N N N~ R, R R R R N N N N N N N N N HN R7~L( R7 HN 7 HN

R R8 R R8 BR N N4N & N N N N N N R NH N N N LKINH HN HN

188

Oct 2023 R8 R9 R8 R R - R - OH R R N N R¹. R!N R¹. RNN R99 N N R² R2 2 R²

2023248067 10

R8 F

R7 o R o R³ O R/ O'R3R3 R O R R F

N N R¹ R9 R¹. R9 N R2 N R² RR²

N - 8 R N R8 R1 OH OH R N.'R NN R o o R³ R R9 R N R¹. RN R 0R99 RN N RR0 N N R¹ N R² 8 R R NHN N R N R R R 10R a R¹ 1 R R¹ 0 R NN N N~ R N N N$ R! N N J' R N N R\_N N RRR N N N R¹ R¹ NH 1 H R10 R 189 N N R R R R9 R - -R R R¹ N R 8 R¹ R R N N R N NNN NLJR N NR N R1 R¹ R¹1 KN H \ /e `R¹

NR NH R N N R8R R R77 ~ R N R R N N R R N R¹ N N N N N R¹ F N N R9 R N R9

N R R ,

wherein: wherein:

R', R 2and R¹, R² 3 independently selected from H, optionally substituted alkyl, methyl, ethyl, andR³Rare are independently selected from H, optionally substituted alkyl, methyl, ethyl, 2-fluoroethyl and 2-fluoroethyl and 2,2,2-trifluoroethyl; 2,2,2-trifluoroethyl; and and

R',R,R', R'and and 1 0 are R¹Rare Ito8 8substituents independently R, , R 1 to substituents independently selectedfrom selected from H,H, optionally optionally

substituted substituted alkyl, alkyl,haloalkyl, haloalkyl,halogen, hydroxyl, halogen, hydroxyl,alkoxy, amino, alkoxy, amino,dialkylamino, dialkylamino,aceylamino, aceylamino,

189

trifluoromethyl or cyano, trifluoromethyl or and wherein cyano, and whereinthe thePTM PTMis is coupled coupled to to a aULM (VLM ULM (VLM or CLM) or CLM) via via L. L.

103961

[0396] In In any any aspect aspect or orembodiment describedherein, embodiment described herein,PTM PTMis is represented represented byby chemical chemical

structure: structure:

N N N N- N N IZ 2023248067 HN N N N H H N N N N N N F

N N N N N N FF N N N F N FF N N F F F F N- N-N F FE N F F N N N_ F F F N F N N N N NN NN N N N N N N II N N IZ N N N N H/ N H N N OH N OH N OH OH N OH N OH F F N N N N NF N N N F N N OH F N F N F OHHF F N OH N OH F FEF N OH N OH F F F F N N- F N/ N N- N FF F F

N N ~ ~N N N N

N OH N HN N OH N OH

190

N N N N- N N IZ -N -N N N HHI( N N F EL N N F EL )N N F E FEL

N N- N N- 1 NN

NN 'N N F EL N N EL F N F EL \FN EL F N F N N EL N EL FFF EL F E N-. N F E N-.. N-.. F EL N N E E N N F N F N N N N F N F NN F EL EL N N E

N N N N 0 N'""N N NN JNN N N N N EL

N N N EL F EL FE F E

N NN N N N N N NN N N N N2J N NJN N N EL N N EL F El F EL F

F F E F F E N N EF NNN' N N N N N N

F QN'NN N 4

N N N N N N N N N N N N N N N F OH HO N H EL Ho

191 I6I

Oct 2023 F F F N N NN N N N N -N N \NN N N NNC No N NN NH NH N NH NH N -N N NH NH F F 2023248067 10

[0397]

[0397] In In any aspect or any aspect or embodiment describedherein, embodiment described thelinker herein,the attachmentpoint linker attachment to PTM pointto PTM is is

as indicatedbybythethedotted as indicated dotted line: line:

8 8 R R 7 N. N R R R R R R¹ R9 N N Z-/ R R N N N N R9 N N Z R¹ N N N MeN Z=N,CH Z=N, CH

CN NR9N N N H2 N.. N N N N N N N N IZ '

, N NLa~j N N H N N N

, N N Me N 2 MeN

[0398] Therapeutic

[0398] Compositions TherapeuticCompositions Pharmaceutical

[0399] Pharmaceutical

[0399] compositions compositions comprising comprising combinations combinations of an effective amount ofamount of an effective at of at least least one one bifunctional bifunctionalcompound as described compound as described herein, herein,and and one one or or more of the more of the compounds compounds otherwise describedherein, otherwise described herein,all all inin effective effective amounts, amounts,inincombination combination withwith a pharmaceutically a pharmaceutically

effective effective amount amount ofofa acarrier, carrier, additive additive or or excipient, excipient, represents represents aa further further aspect aspect ofofthe the present present disclosure. disclosure.

[0400] The present

[0400] The present disclosure disclosure includes, includes, wherewhere applicable, applicable, the compositions the compositions comprising comprising the the pharmaceutically acceptablesalts, pharmaceutically acceptable salts, ininparticular, particular, acid acid or or base baseaddition additionsalts salts ofofcompounds compoundsas as

described herein. described herein. TheThe acids acids which which are used are used to prepare to prepare the pharmaceutically the pharmaceutically acceptable acceptable acid acid addition salts salts of of the the aforementioned basecompounds aforementioned base compounds useful useful according according to this to this aspect aspect are are those those

which form which formnon-toxic non-toxic acid acid addition addition salts, salts, i.e.,salts i.e., saltscontaining containingpharmacologically pharmacologically acceptable acceptable

192 anions, suchasasthethe hydrochloride, hydrobromide, hydroiodide, nitrate, nitrate, 2023248067 10 Oct 2023 anions, such hydrochloride, hydrobromide, hydroiodide, sulfate,sulfate, bisulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, maleate, fumarate, gluconate, gluconate, saccharate, saccharate, benzoate, benzoate, methanesulfonate, methanesulfonate, ethanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate benzenesulfonate, p-toluenesulfonate and pamoate and pamoate [i.e., [i.e., 1,1'-methylene-bis-(2-hydroxy-3 1,1'-methylene-bis-(2-hydroxy-3 naphthoate)]salts, naphthoate)]salts, among numerous among numerous others. others.

Pharmaceuticallyacceptable

[0401] Pharmaceutically

[0401] base acceptablebase addition saltsmaymay additionsalts also also be used be used to produce to produce

pharmaceutically acceptable pharmaceutically acceptablesalt salt forms forms ofofthe the compounds compounds or or derivativesaccording derivatives according to to thethe present present

disclosure. The disclosure. Thechemical chemical bases bases that that may may be beasused used as reagents reagents to pharmaceutically to prepare prepare pharmaceutically acceptable base salts acceptable base salts of of the the present present compounds thatare compounds that are acidic acidic in in nature nature are are those those that that form form non non-

toxic base toxic salts with base salts with such compounds.Such such compounds. Such non-toxic non-toxic basebase salts salts include, include, butbut areare notnot limited limited to to

those derived those derived from fromsuch suchpharmacologically pharmacologically acceptable acceptable cations cations such such as alkali as alkali metal metal cations cations (eg., (eg.,

potassiumand potassium andsodium) sodium) and and alkaline alkaline earth earth metal metal cations cations (eg, (eg, calcium, calcium, zinc magnesium), zinc and and magnesium), ammonium ammonium or water-soluble or water-soluble amine amine addition addition saltssalts suchsuch as N-methylglucamine-(meglumine), as N-methylglucamine-(meglumine) and and the lower the alkanolammonium lower alkanolammonium and and other other basebase salts salts of of pharmaceutically pharmaceutically acceptable acceptable organic organic amines, amines,

among others. among others.

[0402] TheThe

[0402] compounds compounds as described as described herein herein may, may, in accordance in accordance with with the disclosure, the disclosure, be be administered in single administered in single or or divided divided doses doses by by the the oral, oral, parenteral parenteral or or topical topicalroutes. routes. Administration Administration

of the active of the active compound compoundmaymay range range fromfrom continuous continuous (intravenous (intravenous drip)drip) to several to several oraloral

administrations perday administrations per day(for(for example, example, Q.I.D.) Q.I.D.) andinclude and may may oral, include oral, parenteral, topical, topical, parenteral, intramuscular, intravenous, sub-cutaneous, intramuscular, intravenous, sub-cutaneous, transdermal transdermal (which (which may ainclude may include a penetration penetration

enhancement agent),buccal, enhancement agent), buccal,sublingual sublingualandand suppository suppository administration, administration, among among other other routes routes of of administration. Enteric administration. Enteric coated coatedoral oraltablets tablets may mayalso alsobebeused used to to enhance enhance bioavailability bioavailability of the of the

compoundsfrom compounds from an oral an oral route route of administration. of administration. The The most most effective effective dosage dosage formdepend form will will depend upon the pharmacokinetics of the particular agent chosen as well as the severity of disease in the upon the pharmacokinetics of the particular agent chosen as well as the severity of disease in the

patient. Administration patient. Administrationofofcompounds compounds according according to present to the the present disclosure disclosure as sprays, as sprays, mists, mists, or or aerosols for aerosols for intra-nasal, intra-nasal, intra-tracheal intra-trachealor or pulmonary administrationmaymay pulmonary administration alsoalso be used. be used. The The present disclosure present disclosure therefore therefore also alsoisisdirected directedtotopharmaceutical pharmaceutical compositions compositions comprising comprising an an effective effective amount of compound amount of compound as described as described herein, herein, optionallyin in optionally combination combination withwith a a pharmaceutically acceptable pharmaceutically acceptablecarrier, carrier, additive additive or or excipient. Compounds excipient. Compounds according according to the to the present present

disclosureion may disclosureion maybebeadministered administered in immediate in immediate release, release, intermediate intermediate release release or sustained or sustained or or controlled release forms. controlled release forms. Sustained Sustained or controlled or controlled release release forms forms are preferably are preferably administered administered

193 orally, orally, but but also alsoin suppositoryand insuppository and transdermal transdermal or other topical forms. other topical Intramuscularinjections injections 2023248067 10 Oct 2023 forms. Intramuscular in liposomal formmay liposomal form mayalso alsobebeused usedtotocontrol controlororsustain sustain the the release release of compound compound atatananinjection injection site. site.

[0403] TheThe

[0403] as as compositions compositions described hereinmay describedherein conventional manner formulatedinin aa conventional maybebeformulated manner

using one or using one or more more pharmaceutically pharmaceutically acceptable acceptable carriers carriers and and may also be may also be administered administered in in controlled-release controlled-release formulations. formulations. Pharmaceutically acceptablecarriers Pharmaceutically acceptable carriers that that may maybebeused used in in these these

pharmaceutical compositions pharmaceutical compositionsinclude, include,but butare arenot notlimited limitedto, to, ion ion exchangers, exchangers, alumina, alumina,aluminum aluminum stearate, stearate, lecithin, lecithin,serum proteins, such serum proteins, such as as human human serum serum albumin, albumin, bufferbuffer substances substances such assuch as

phosphates, glycine, phosphates, glycine, sorbic sorbicacid, acid,potassium potassium sorbate, sorbate, partial partial glyceride glyceride mixtures mixtures of saturated of saturated

vegetable fatty acids, vegetable fatty acids, water, water, salts saltsororelectrolytes, such electrolytes, as as such prolamine prolaminesulfate, sulfate,disodium disodiumhydrogen hydrogen

phosphate, potassium phosphate, potassium hydrogen hydrogen phosphate, phosphate, sodium chloride, sodium chloride, zinc zinc salts, salts, silica, colloidal colloidal silica, magnesium magnesium trisilicate, polyvinyl trisilicate, polyvinyl pyrrolidone, pyrrolidone,cellulose-based cellulose-basedsubstances, substances,polyethylene polyethylene glycol, glycol,

sodium carboxymethylcellulose, sodium carboxymethylcellulose, polyacrylates, polyacrylates, waxes, waxes, polyethylene-polyoxypropylene-block polyethylene-polyoxypropylene-block

polymers, polyethylene polymers, polyethyleneglycol glycoland andwool woolfat. fat.

[0404] The compositions

[0404] The compositions as described as described herein herein may be administered may be administered orally, parenterally, orally, parenterally, by by inhalation spray,topically, inhalation spray, topically, rectally, rectally, nasally, nasally, buccally, buccally, vaginally vaginally or implanted or via an via an implanted reservoir. reservoir.

The term The term"parenteral" "parenteral"asasused usedherein hereinincludes includessubcutaneous, subcutaneous, intravenous, intravenous, intramuscular, intramuscular, intra intra-

articular, intra-synovial, articular, intra-synovial,intrasternal, intrasternal, intrathecal, intrathecal, intrahepatic, intrahepatic, intralesional intralesional and intracranial and intracranial

injection or infusion injection or infusiontechniques. techniques. Preferably, Preferably, the compositions the compositions are administered are administered orally, orally, intraperitoneally intraperitoneally oror intravenously. intravenously.

[0405] Sterile

[0405] Sterile injectable injectable forms forms of the of the compositions compositions as described as described may bemay hereinherein be aqueous aqueous or or oleaginous suspension. oleaginous suspension. These Thesesuspensions suspensions maymay be formulated be formulated according according to techniques to techniques knownknown in in the art the art using using suitable suitable dispersing dispersing or orwetting wetting agents agents and suspending agents. and suspending agents. The Thesterile sterile injectable injectable preparation may preparation mayalso alsobebea asterile sterile injectable injectable solution solution or or suspension suspensioninina anon-toxic non-toxicparenterally- parenterally acceptable diluent diluent or or solvent, solvent,for forexample example as as aa solution solutioninin1,1, 3-butanediol. Among 3-butanediol. Among the acceptable acceptable

vehicles and vehicles and solvents solvents that that may maybebe employed employed are water, are water, Ringer's Ringer's solution solution and isotonic and isotonic sodium sodium

chloride solution. solution. In addition, addition, sterile, sterile, fixed fixedoils oilsare areconventionally conventionally employed employed asasa asolvent solventor or suspending medium. suspending medium. ForFor this this purpose, purpose, anyany bland bland fixed fixed oiloil maymay be employed be employed including including synthetic synthetic

mono-or or mono- di-glycerides. di-glycerides. Fatty Fatty acids, acids, such such as as acid oleic oleicandacid its and its glyceride glyceride derivatives derivatives are useful are in useful in the preparation the preparationof of injectables, injectables, as natural as are are natural pharmaceutically-acceptable pharmaceutically-acceptable oils, oils, such as such olive oil as olive oil

194 or castor castor oil, oil, especially especially in intheir theirpolyoxyethylated polyoxyethylated versions. versions. These oil solutions solutions or or suspensions suspensions 2023248067 10 Oct 2023

These oil

mayalso may alsocontain contain a long-chain a long-chain alcohol alcohol diluentdiluent or dispersant, or dispersant, such as such as Ph. Ph. Helv Helv or or similar similar alcohol. alcohol.

The pharmaceutical

[0406] The pharmaceutical

[0406] compositions compositions as described as described herein herein may be orally orally administered may beadministered in in any orally acceptable any orally acceptabledosage dosage form form including, including, but limited but not not limited to, capsules, to, capsules, tablets, tablets, aqueous aqueous

suspensions oror solutions. suspensions solutions. In In the the case case of of tablets tablets for for oral oral use, use,carriers carrierswhich which are arecommonly used commonly used

include lactose lactose and andcorn corn starch.Lubricating starch. Lubricating agents, agents, such such as magnesium as magnesium stearate,stearate, are alsoare also

typically added. For typically For oral oral administration administration inin aa capsule capsule form, form,useful usefuldiluents diluentsinclude includelactose lactoseand and dried corn dried corn starch. starch. When aqueous When aqueous suspensions suspensions are are required required for for oraloral use, use, thethe active active ingredient ingredient is is combinedwith combined with emulsifying emulsifying andand suspending suspending agents. agents. If desired, If desired, certain certain sweetening, sweetening, flavoring flavoring or or coloring coloring agents mayalso agents may also be be added. added.

[0407] Alternatively,

[0407] Alternatively, the thepharmaceutical pharmaceuticalcompositions compositions as described as described herein herein may may be be administered in the administered in the form formofofsuppositories suppositoriesforforrectal rectaladministration. administration.These Thesecancan be be prepared prepared by by

mixing the mixing the agent agent with witha asuitable suitable non-irritating non-irritating excipient, excipient,which which is is solid solidatatroom room temperature temperature but

liquid at rectal liquid at rectal temperature andtherefore temperature and thereforewill willmelt meltin inthethe rectum rectum to release to release the the drug. drug. Such Such

materials include cocoa materials butter, beeswax cocoa butter, andpolyethylene beeswax and polyethyleneglycols. glycols.

[0408] TheThe

[0408] pharmaceutical pharmaceutical compositions compositions as described as described herein herein alsoalso maymay be administered be administered

topically. Suitable topically. Suitable topical topical formulations formulations are are readily readily prepared for each prepared for each of of these these areas areas oror organs. organs. Topical application Topical application for forthethelower lower intestinal intestinal tract tract can can be effected be effected in a rectal in a rectal suppository suppository

formulation (see above) formulation (see above)ororinina asuitable suitableenema enema formulation. formulation. Topically-acceptable Topically-acceptable transdermal transdermal

patches may patches mayalso also be beused. used. For topical

[0409] For topical

[0409] applications, applications, the pharmaceutical the pharmaceutical compositions compositions may be formulated may be formulated in a in a suitable ointment containing suitable ointment containingthetheactive activecomponent component suspended suspended or dissolved or dissolved in more in one or one or more carriers. carriers.Carriers Carriersfor fortopical administration topical administrationofofthe thecompounds ofthis compounds of this invention invention include, include, but are are not limited not limited to, to, mineral mineraloil, oil, liquid liquidpetrolatum, petrolatum,white white petrolatum, petrolatum, propylene propylene glycol, glycol,

polyoxyethylene, polyoxypropylene polyoxyethylene, polyoxypropylene compound, compound, emulsifying emulsifying wax wax and and water. water. In certain In certain preferred preferred

aspects of of the the invention, invention, the the compounds compounds maymay be coated be coated onto onto a stent a stent whichwhich is to is be to be surgically surgically

implanted into implanted into a patient a patient in order in order to inhibit to inhibit or reduce or reduce the likelihood the likelihood of occlusion of occlusion occurring occurring in the in the stent in the stent in the patient. patient. the the Alternatively,

[0410] Alternatively,

[0410] pharmaceutical pharmaceutical compositions can becan compositions be formulated formulated in a suitable lotionlotion in a suitable or cream containing or cream containing the theactive activecomponents components suspended suspended or dissolved or dissolved in orone in one moreor more

pharmaceutically pharmaceutically acceptable acceptable carriers. carriers. Suitable Suitable carriers carriers include, include, but are but not are not to, limited mineral limited to,oil, mineral oil,

195 sorbitan monostearate, polysorbate polysorbate60,60, 2023248067 10 Oct 2023 sorbitan monostearate, cetyl cetyl esters esters wax, wax, cetearyl cetearyl alcohol, alcohol, 2-octyldodecanol, 2-octyldodecanol, benzyl alcohol benzyl alcohol and and water. water.

[0411] ForFor

[0411] use,use, ophthalmic ophthalmic the pharmaceutical the pharmaceutical compositions compositions may may be be formulated formulated as as micronized suspensions micronized suspensionsin in isotonic,pHpH isotonic, adjusted adjusted sterile sterile saline,or,or,preferably, saline, preferably,as assolutions solutionsin in isotonic, isotonic, pH adjusted sterile pH adjusted sterile saline, saline, either either with our without with our without a apreservative preservative such suchas as benzylalkoniumchloride. benzylalkonium chloride.Alternatively, Alternatively,for forophthalmic ophthalmic uses, uses, thethe pharmaceutical pharmaceutical compositions compositions

maybebeformulated may formulatedininananointment ointmentsuch suchasaspetrolatum. petrolatum.

[0412] TheThe

[0412] pharmaceutical pharmaceutical compositions compositions as as describedherein described herein may mayalso also be be administered administered by nasal aerosol or nasal or inhalation. inhalation.Such Such compositions are prepared compositions are prepared according accordingtototechniques techniqueswell-known well-known in the the art artof ofpharmaceutical pharmaceutical formulation andmay formulation and maybebe prepared prepared as as solutions solutions in in saline,employing saline, employing benzyl alcohol benzyl alcohol or or other other suitable suitable preservatives, preservatives, absorption promoterstotoenhance absorption promoters enhancebioavailability, bioavailability, fluorocarbons, and/or fluorocarbons, and/or other other conventional conventional solubilizing solubilizing or dispersing or dispersing agents. agents.

[0413] TheThe

[0413] amount amount of compound in a in of compound a pharmaceutical pharmaceutical composition as as composition describedherein described that herein that maybebecombined may combined with with thethe carriermaterials carrier materialstotoproduce producea asingle singledosage dosageform form willvary will varydepending depending uponthe upon thehost hostandand disease disease treated, treated, the particular the particular mode mode of administration. of administration. Preferably, Preferably, the the compositions should be compositions should formulated to be formulated to contain contain between about 0.05 between about 0.05 milligram milligram to to about about 750 750 milligrams or milligrams or more, more, more morepreferably preferablyabout about1 milligram 1 milligram to to about about 600600 milligrams, milligrams, and and eveneven more more

preferably about preferably about1010milligrams milligrams to about to about 500 milligrams 500 milligrams of ingredient, of active active ingredient, alone or alone in or in combinationwith combination withatatleast least one other other compound according compound according to to thepresent the presentdisclosure. disclosure.

[0414] It should

[0414] It should also also be understood be understood that athat a specific specific dosagedosage and treatment and treatment regimen regimen for any for any particular patient particular patient will will depend upona avariety depend upon varietyofoffactors, factors, including includingthe theactivity activityofofthe thespecific specific compound employed, compound employed, the the age,age, body body weight, weight, general general health, health, sex,sex, diet,time diet, timeofof administration,rate administration, rate of excretion, excretion, drug combination,and drug combination, andthe thejudgment judgment of the of the treating treating physician physician andand the the severity severity of of

the particular the particulardisease diseaseororcondition condition being being treated. treated.

[0415] A patient

[0415] A patient or subject or subject in need in need of therapy using using of therapy compounds compounds according according to the to the methods methods described herein described herein can can be be treated treated by by administering administeringtotothe thepatient patient (subject) (subject) an an effective effective amount of amount of

the compound the according compound according to the to the present present disclosure disclosure including including pharmaceutically pharmaceutically acceptable acceptable salts, salts,

solvates solvates or polymorphs, thereofoptionally polymorphs, thereof optionallyinina apharmaceutically pharmaceutically acceptable acceptable carrier carrier or or diluent, diluent,

either either alone, alone, or or in in combination with other combination with other known known erythopoiesisstimulating erythopoiesis stimulating agents agents as as otherwise otherwise

identified herein. identified herein.

196

[0416] TheseThese

[0416] compounds compounds can be administered can be administered by any appropriate by any appropriate route, route, for for example, example, orally, orally, parenterally,intravenously, parenterally, intravenously, intradermally, intradermally, subcutaneously, subcutaneously, or topically, or topically, including including transdermally, transdermally,

in liquid, cream, in liquid, cream,gel, gel,ororsolid solidform, form, or or by by aerosol aerosol form.form.

[0417] The active

[0417] The active compound compound is included in the in is included the pharmaceutically pharmaceutically acceptable carriercarrier acceptable or diluent or diluent

in an amount in an amount sufficient sufficient to deliver to deliver to a to a patient patient a therapeutically a therapeutically effective effective amount amount for for the desired the desired

indication, indication, without causing serious without causing serious toxic toxic effects effects in the the patient patient treated. treated.AA preferred preferred dose of the dose of

active active compound forall compound for allofofthe the herein-mentioned herein-mentionedconditions conditionsis isininthe therange rangefrom fromabout about 10 10 ng/kg ng/kg

to 300 to 300 mg/kg, mg/kg,preferably preferably0.10.1to to100100 mg/kg mg/kg per day, per day, more generally more generally 0.5 to 0.5 to 25 about about 25 mg per mg per kilogram body kilogram bodyweight weight of of therecipient/patient the recipient/patient per perday. day. AAtypical typicaltopical topical dosage dosagewill willrange rangefrom from 0.01-5% wt/wt 0.01-5% wt/wt in ain a suitable suitable carrier. carrier.

[0418] TheThe

[0418] compound compound is conveniently is conveniently administered administered in suitable in any any suitable unit unit dosage dosage form,form,

including but but not not limited to to one one containing containing less less than than 1mg, 1mg, 11 mg to 3000 mg to 3000mg, mg,preferably preferably5 5toto500 500 mgofofactive mg activeingredient ingredientperperunit unitdosage dosage form. form. An oral An oral dosage dosage of 25-250 of about about 25-250 mg mg is often is often convenient. convenient.

[0419] TheThe

[0419] active active ingredient ingredient is preferably is preferably administered administered to achieve to achieve peak peak plasma plasma concentrations of the concentrations of the active active compound compound of of about about 0.00001-30 0.00001-30 mM, preferably mM, preferably about 0.1-30 about 0.1-30 µM. [M. This may maybebeachieved, achieved,for forexample, example,by by thethe intravenous intravenous injectionof of injection a a solutionororformulation solution formulationofof the active the active ingredient, ingredient, optionally optionally in in saline, saline,ororananaqueous aqueous medium medium ororadministered administeredasasa abolus bolusofof the active the active ingredient. ingredient. Oral Oraladministration administrationis isalso also appropriate appropriate to generate to generate effective effective plasma plasma

concentrations concentrations of of active active agent. agent.

[0420] TheThe

[0420] of of concentration concentration active compound activecompound in the in the drugdrug composition willwill composition depend depend on on absorption, distribution, inactivation, absorption, distribution, inactivation,and and excretion excretion rates rates of the drug as the drug as well wellasasother otherfactors factors knowntotothose known thoseofofskill skillinin the the art. art. It It isis totobe be noted noted that that dosage values will dosage values will also also vary vary with withthe the severity severity ofofthe thecondition condition toalleviated. to be be alleviated. It is It to is be to be further further understood understood that that for any for any particular particular

subject, subject, specific specific dosage dosage regimens shouldbebeadjusted regimens should adjustedover overtime timeaccording accordingtotothe theindividual individualneed need and the professional and the professional judgment judgmentofofthe theperson personadministering administering or or supervising supervising thethe administration administration of of

the compositions, the andthat compositions, and that the the concentration concentration ranges rangesset set forth forth herein herein are are exemplary exemplaryonly onlyand and areare

not intended not intended to to limit limit the thescope scope or orpractice practiceofof thethe claimed claimedcomposition. composition. The The active active ingredient ingredientmay may

be administered be administered at at once, once, or or may bedivided may be dividedinto intoaanumber numberofof smallerdoses smaller dosestotobebeadministered administered at at

varying intervalsofof varying intervals time. time.

197

[0421] Oral Oral

[0421] compositions compositions will generally will generally include include an diluent an inert inert diluent or an or an edible edible carrier. They carrier. They maybebeenclosed may enclosed in gelatin in gelatin capsules capsules or compressed or compressed into tablets. into tablets. For the For the of purpose purpose oral of oral therapeutic administration, the therapeutic administration, the active active compound compound or or itsits prodrug prodrug derivative derivative cancan be incorporated be incorporated

with excipients with excipients and andusedused in form in the the of form of tablets, tablets, troches, troches, or capsules. or capsules. Pharmaceutically Pharmaceutically

compatible bindingagents, compatible binding agents, and/or and/or adjuvant adjuvantmaterials materials can canbe beincluded includedasas part part of of the composition. composition.

[0422] The tablets,

[0422] The tablets, pills, pills, capsules, capsules, troches troches and and the like the like can contain can contain any ofany the of the following following

ingredients, ingredients, or or compounds compounds ofof a asimilar similarnature: nature: a abinder bindersuch suchasasmicrocrystalline microcrystallinecellulose, cellulose, gum gum tragacanth or tragacanth or gelatin; gelatin; an an excipient such as excipient such as starch starch or or lactose, lactose, aa dispersing dispersing agent such such as as alginic alginic acid, Primogel, acid, Primogel,or or corn corn starch; starch; a lubricant a lubricant such such as magnesium as magnesium stearate stearate or or aSterotes; Sterotes; a glidant glidant such such as colloidal silicon as colloidal silicondioxide; dioxide;a sweetening a sweetening agentagent such such as as sucrose sucrose or saccharin; or saccharin; or a flavoring or a flavoring agent agent such as peppermint, such as peppermint,methyl methyl salicylate,or or salicylate, orange orange flavoring. flavoring. When When the dosage the dosage unitisform unit form a is a capsule, capsule, itit can cancontain, contain,in in addition addition to material to material of theofabove the above type, a type, liquid acarrier liquid such carrier as a such fatty as a fatty

oil. oil.In Inaddition, addition,dosage dosageunit unitforms formscan cancontain containvarious variousother othermaterials materialswhich which modify the physical modify the physical

form form ofof thedosage the dosage unit, unit, for for example, example, coatings coatings of sugar, of sugar, shellac,shellac, or enteric or enteric agents. agents.

The active

[0423] The active

[0423] compound compound or pharmaceutically or pharmaceutically acceptable acceptable salt thereof can be can salt thereof be administered administered

as aa component component of of anan elixir, suspension, elixir, suspension,syrup, syrup,wafer, wafer,chewing chewing gum gum or like. or the the like. A syrup A syrup may may

contain, in addition contain, in addition totothe theactive activecompounds, compounds, sucrose sucrose as a sweetening as a sweetening agent andagent and certain certain

preservatives,dyes preservatives, dyes andand colorings colorings and flavors. and flavors.

The active

[0424] The active

[0424] compound compound or pharmaceutically or pharmaceutically acceptable acceptable salts thereof salts thereof canbealso can also be mixed mixed with other with otheractive activematerials materialsthat thatdo do not not impair impair the desired the desired action,action, or withormaterials with materials that that supplementthe supplement thedesired desiredaction, action,such such as as erythropoietin erythropoietin stimulating stimulating agents, agents, including including EPO EPO and and darbapoietin alfa, darbapoietin alfa, among amongothers. others.In In certainpreferred certain preferred aspects aspects of the of the invention, invention, one one or or more more compoundsaccording compounds according to to thethe present present disclosurearearecoadministered disclosure coadministered with with another another bioactive bioactive agent, agent,

such as an such as an erythropoietin erythropoietin stimulating stimulating agent agent or or aa would healingagent, would healing agent, including includingananantibiotic, antibiotic, as as otherwise described herein. otherwise described herein.

[0425] Solutions

[0425] Solutions or suspensions or suspensions usedparenteral, used for for parenteral, intradermal, intradermal, subcutaneous, subcutaneous, or topical or topical

application can application can include include the the following followingcomponents: components: a sterilediluent a sterile diluentsuch suchasaswater waterforforinjection, injection, saline saline solution, solution, fixed fixed oils, oils,polyethylene polyethylene glycols, glycols, glycerine, glycerine,propylene glycol or propylene glycol or other other synthetic synthetic solvents; solvents; antibacterial antibacterial agents agents such as benzyl such as alcohol orormethyl benzyl alcohol methylparabens; parabens;antioxidants antioxidants such such as as

ascorbic acid ascorbic acid ororsodium sodium bisulfite;chelating bisulfite; chelatingagents agents suchsuch as ethylenediaminetetraacetic as ethylenediaminetetraacetic acid; acid; buffers such buffers suchas as acetates, acetates, citrates citrates or phosphates or phosphates and agents and agents for the for the adjustment adjustment of tonicityof tonicity such as such as

198 sodium chlorideorordextrose. dextrose. The Theparental parentalpreparation preparationcancanbebeenclosed enclosed in in ampoules, disposable 2023248067 10 Oct 2023 sodium chloride ampoules, disposable syringes syringes orormultiple multiple dose dose vials vials mademade of glass of glass or plastic. or plastic.

[0426] If administered

[0426] If administered intravenously, intravenously, preferred preferred carriers carriers areare physiological physiological saline saline or or phosphate phosphate

buffered saline buffered saline (PBS). (PBS).

[0427] one embodiment, In embodiment,

[0427] In one the active the active compounds compounds are prepared are prepared with carriers with carriers that will will protect thatprotect the compound the againstrapid compound against rapidelimination eliminationfrom from thethe body, body, such such as as a controlledrelease a controlled releaseformulation, formulation, including implantsandand including implants microencapsulated microencapsulated delivery delivery systems. systems. Biodegradable, Biodegradable, biocompatible biocompatible

polymerscancanbe be polymers used, used, such such as ethylene as ethylene vinyl acetate, vinyl acetate, polyanhydrides, polyanhydrides, polyglycolic polyglycolic acid, acid, collagen, collagen, polyorthoesters, polyorthoesters, and and polylactic polylactic acid. acid.Methods for preparation Methods for preparation of of such such formulations will formulations will

be apparent to those skilled in the art. be apparent to those skilled in the art.

[0428] Liposomal

[0428] Liposomal suspensions suspensions may may also be also be pharmaceutically pharmaceutically acceptable acceptable carriers.carriers. These These may may be prepared be prepared according accordingtotomethods methodsknown known to those to those skilled skilled in in theart, the art, for for example, example,asasdescribed describedinin U.S. Pat. U.S. Pat. No. 4,522,811 (which No. 4,522,811 (whichisisincorporated incorporatedherein hereinbybyreference referenceininits its entirety). entirety). For For example, example,

liposome formulationsmaymay liposome formulations be prepared be prepared by dissolving by dissolving appropriate appropriate lipid(s) lipid(s) (such (such as as stearoyl stearoyl

phosphatidyl ethanolamine, phosphatidyl ethanolamine,stearoyl stearoylphosphatidyl phosphatidylcholine, choline,arachadoyl arachadoylphosphatidyl phosphatidyl choline, choline, andand

cholesterol) cholesterol) in in an an inorganic inorganic solvent that isis then solvent that then evaporated, evaporated, leaving leaving behind behind aa thin thin film film of of dried dried lipid lipid on the surface on the surface ofofthe the container. container. AnAnaqueous aqueous solution solution of the of the active active compound compound are then are then

introduced into the introduced into the container. container. The container is The container is then then swirled swirled by by hand handtotofree free lipid lipid material material from from the sides the sides of of the the container containerandand to to disperse disperse lipid lipid aggregates, aggregates, thereby thereby forming forming the liposomal the liposomal

suspension. suspension.

[0429] Therapeutic

[0429] Methods TherapeuticMethods

[0430] In anInadditional

[0430] an additional aspect, aspect, the the description description provides provides therapeutic therapeutic compositions compositions comprising comprising

an effective amount an effective ofa acompound amount of compound as described as described herein herein or salt or salt form form thereof, thereof, and aand a

pharmaceutically acceptable pharmaceutically acceptablecarrier. carrier. The Thetherapeutic therapeutic compositions compositionsmodulate modulate protein protein degradation degradation

in in aa patient patient or orsubject, subject,for forexample, example, an an animal animal such as aa human, such as andcan human, and canbebeused usedforfortreating treatingoror ameliorating disease ameliorating disease states states or orconditions conditions which which are are modulated throughthe modulated through thedegraded degradedprotein. protein. The terms

[0431] The terms

[0431] "treat", "treat", "treating", and and "treating", "treatment", "treatment", etc., etc., as as used used herein, anyaction refertoto any herein,refer action providing aa benefit providing benefit to to aapatient patientfor forwhich whichthe thepresent presentcompounds maybebeadministered, compounds may administered,including including the treatment the treatment of of any disease state any disease state or or condition condition which is modulated which is throughthe modulated through theprotein proteintotowhich which the present the present compounds compounds bind. bind. Disease Disease statesstates or conditions, or conditions, including including cancer, cancer, which which may be may be treated using treated using compounds accordingtotothethepresent compounds according presentdisclosure disclosureare are set set forth forth hereinabove. hereinabove.

199 herein herein as described for 2023248067 10 Oct 2023

[0432] TheThe

[0432] description description provides provides therapeutic therapeutic compositions compositions as described for

effectuatingthe effectuating thedegradation degradation of proteins of proteins of interest of interest fortreatment for the the treatment or amelioration or amelioration of a disease, of a disease, e.g., cancer. e.g., cancer. In certain certain additional additional embodiments, thedisease embodiments, the diseaseisis multiple multiple myeloma. myeloma. As such, As such, in in anotheraspect, another aspect,thethedescription description provides provides a method a method of ubiquitinating/ of ubiquitinating/ degrading degrading a target a target protein in protein in a cell. cell.InIncertain certainembodiments, embodiments, the the method comprisesadministering method comprises administeringa bifunctional a bifunctionalcompound compound as as

described herein described herein comprising, comprising, e.g., e.g., aa CLM anda aPTM, CLM and PTM, preferably preferably linked linked through through a linker a linker moiety, moiety,

as otherwise described described herein, herein, wherein whereinthe theCLM CLM is coupled is coupled to the to the PTM PTM and wherein and wherein the CLMthe CLM

recognizes aa ubiquitin recognizes ubiquitin pathway pathway protein protein (e.g.,an an (e.g., ubiquitin ubiquitin ligase,preferably ligase, preferably an an E3 ubiquitin E3 ubiquitin

ligase suchas, ligase such as,e.g., e.g., cereblon) cereblon)andand the the PTM PTM recognizes recognizes theprotein the target target such protein that such that degradation degradation of of the target the target protein proteinwill willoccur occur when when the target the target protein protein is placed is placed in proximity in proximity to the ubiquitin to the ubiquitin ligase, ligase, thus resulting thus resulting in in degradation/inhibition degradation/inhibition of of the the effects effects of of the the target target protein protein and and the the control control ofof proteinlevels. protein levels. The The control control of protein of protein levels levels afforded afforded by theby the present present disclosure disclosure provides provides treatment treatment of aa disease state state or or condition, condition, which is modulated which is modulatedthrough through thethe targetprotein target proteinbyby lowering lowering the the

level of that level of that protein in the protein in cell, e.g., the cell, e.g.,cell cellofofa apatient. patient. In In certain certain embodiments, themethod embodiments, the method comprises administeringan an comprises administering effectiveamount effective amount of a of a compound compound as described as described herein, herein, optionally optionally

including aa pharamaceutically pharamaceutically acceptable acceptableexcipient, excipient, carrier, carrier, adjuvant, adjuvant, another bioactive agent or or combinationthereof. combination thereof.

[0433] In In

[0433] additionalembodiments, additional embodiments, the the description description provides provides methods methods for treating for treating or or emelioratinga disease, emeliorating a disease, disorder disorder or symptom or symptom thereof thereof in a or in a subject subject or a e.g., a patient, patient, e.g., an an animal animal such such as as a a human, comprisingadministering human, comprising administering to to a subjectininneed a subject needthereof thereof a a composition composition comprising comprising an an

effective amount, effective e.g., aa therapeutically amount, e.g., therapeuticallyeffective effectiveamount, amount, of ofa acompound as described compound as describedherein hereinoror salt salt form thereof, and form thereof, anda pharmaceutically a pharmaceutically acceptable acceptable excipient, excipient, carrier, carrier, adjuvant, adjuvant, anotheranother

bioactive agent bioactive agent ororcombination combination thereof,wherein thereof, wherein the the composition composition is effective is effective for treating for treating or or amelioratingthethe ameliorating disease disease or disorder or disorder or symptom or symptom thereof thereof in the in the subject. subject.

[0434] In another

[0434] In another aspect, aspect, the the description description provides provides methods methods for identifying for identifying the effects the effects of of the the degradation ofofproteins degradation proteins ofofinterest interest inin aa biological biological system systemusing using compounds compounds according according to theto the presentdisclosure. present disclosure.

[0435] In another

[0435] In another embodiment, embodiment, the present the present disclosure disclosure is directed is directed to a method to a method of treating of treating a a humanpatient human patientininneed needforfora adisease diseasestate stateororcondition conditionmodulated modulated through through a protein a protein where where the the degradation ofofthat degradation thatprotein proteinwill will produce produce a therapeutic a therapeutic effecteffect in patient, in that that patient, the the method method comprising administering comprising administeringtotoa apatient patient inin need needananeffective effective amount amountof of a compound a compound according according to to

200 the present disclosure, disclosure, optionally optionally in incombination with another another bioactive bioactive agent. agent. The The disease disease state state 2023248067 10 Oct 2023 the combination with or condition may maybebea adisease diseasecaused causedby by a microbial a microbial agent agent or other or other exogenous exogenous agentagent such such as a as a virus, bacteria, virus, bacteria,fungus, fungus,protozoa protozoa or or other other microbe or may microbe or bea adisease may be disease state, state, which is caused which is caused by by overexpression overexpression ofprotein, of a a protein, which which leads leads to a disease to a disease state and/or state and/or condition condition termterm

[0436] TheThe

[0436] "disease "disease state or or state condition" to to used condition"is isused describeanyany describe disease stateoror diseasestate condition wherein whereinprotein proteindysregulation dysregulation(i.e., (i.e., the the amount amountof of protein protein expressed expressed in ainpatient a patient is is

elevated) occurs and elevated) occurs andwhere where degradation degradation of or of one onemore or proteins more proteins in a may in a patient patient may provide provide

beneficial therapy or beneficial or relief relief of of symptoms symptoms totoa apatient patientininneed needthereof. thereof.InIncertain certaininstances, instances, the the disease stateororcondition disease state conditionmaymay be cured. be cured.

Disease

[0437] Disease

[0437] states states of conditions of conditions which which may may be treated using using be treated compounds compounds according according to the to the present disclosure present disclosure include, include, for for example, example, asthma, autoimmunediseases asthma, autoimmune diseases such such as as multiplesclerosis, multiple sclerosis, various cancers, cancers, ciliopathies, ciliopathies, cleft cleft palate, palate, diabetes, diabetes,heart heartdisease, disease,hypertension, hypertension, inflammatory inflammatory

boweldisease, bowel disease, mental mental retardation, retardation, mood disorder, mood disorder, obesity, obesity, refractive refractive error, infertility, error, infertility, Angelman Angelman syndrome, Canavan syndrome, Canavan disease, disease, Coeliac Coeliac disease, disease, Charcot-Marie-Tooth Charcot-Marie-Tooth disease, disease, Cystic fibrosis, Cystic fibrosis,

Duchenne muscular Duchenne musculardystrophy, dystrophy,Haemochromatosis, Haemochromatosis, Haemophilia, Haemophilia, Klinefelter's Klinefelter's syndrome, syndrome,

Neurofibromatosis,Phenylketonuria, Neurofibromatosis, Phenylketonuria,Polycystic Polycystic kidney kidney disease, disease, (PKD1) (PKD1) or 4 (PKD2) or 4 (PKD2) Prader- Prader

Willi syndrome, Willi Sickle-cell disease, syndrome, Sickle-cell disease, Tay-Sachs disease, Turner Tay-Sachs disease, Turnersyndrome. syndrome.

[0438] Further

[0438] Further disease disease states states or conditions or conditions which which may may be treated be treated by compounds by compounds according according to to the present the present disclosure disclosure include include Alzheimer's disease, Amyotrophic Alzheimer's disease, Amyotrophic lateralsclerosis lateral sclerosis (Lou (LouGehrig's Gehrig's disease), Anorexia disease), nervosa,Anxiety Anorexia nervosa, Anxiety disorder, disorder, Atherosclerosis, Atherosclerosis, Attention Attention deficit deficit hyperactivity hyperactivity

disorder, Autism, disorder, Bipolardisorder, Autism, Bipolar disorder, Chronic Chronicfatigue fatiguesyndrome, syndrome, Chronic Chronic obstructive obstructive pulmonary pulmonary

disease, disease, Crohn's Crohn's disease, disease, Coronary heart disease, Coronary heart disease, Dementia, Depression, Diabetes Dementia, Depression, Diabetesmellitus mellitustype type1,1, Diabetes mellitus Diabetes mellitus type type 2, 2, Epilepsy, Epilepsy, Guillain-Barré Guillain-Barr6 syndrome, syndrome,Irritable Irritablebowel bowelsyndrome, syndrome, Lupus, Lupus,

Metabolic syndrome, Metabolic syndrome,Multiple Multiple sclerosis,Myocardial sclerosis, Myocardial infarction,Obesity, infarction, Obesity, Obsessive-compulsive Obsessive-compulsive

disorder, Panic disorder, Panic disorder, disorder,Parkinson's Parkinson'sdisease, disease, Psoriasis, Psoriasis, Rheumatoid Rheumatoid arthritis, arthritis, Sarcoidosis, Sarcoidosis,

Schizophrenia, Stroke, Schizophrenia, Stroke, Thromboangiitis Thromboangiitisobliterans, obliterans, Tourette Tourettesyndrome, syndrome,Vasculitis. Vasculitis.

[0439] Stilladditional

[0439] Still states or disease states additional disease or conditions can be which can conditionswhich treatedbybycompounds be treated compounds

according totothe according thepresent present disclosure disclosure include include aceruloplasminemia, aceruloplasminemia, Achondrogenesis Achondrogenesis type II, type II, achondroplasia, Acrocephaly, Acrocephaly,Gaucher Gaucher disease disease type type 2, acute 2, acute intermittent intermittent porphyria, porphyria, Canavan Canavan

disease, Adenomatous disease, Adenomatous Polyposis Polyposis Coli,Coli, ALA dehydratase ALA dehydratase deficiency, deficiency, adenylosuccinate adenylosuccinate lyase lyase deficiency, deficiency, Adrenogenital syndrome, Adrenoleukodystrophy, Adrenogenital syndrome, Adrenoleukodystrophy,ALA-D ALA-D porphyria, porphyria, ALA ALA

201

Oct 2023 dehydratase deficiency, dehydratase Alkaptonuria,Alexander deficiency, Alkaptonuria, Alexander disease, disease, Alkaptonuric Alkaptonuric ochronosis, ochronosis, alpha alpha 1- 1 antitrypsin antitrypsin deficiency, deficiency, alpha-i proteinase inhibitor, alpha-1 proteinase inhibitor, emphysema, amyotrophic emphysema, amyotrophic lateral lateral sclerosis sclerosis

Alstr6msyndrome, Alström syndrome, Alexander Alexander disease, disease, Amelogenesis Amelogenesis imperfecta, imperfecta, ALA dehydratase ALA dehydratase deficiency, deficiency,

Anderson-Fabry disease, Anderson-Fabry disease, androgen insensitivity syndrome, androgen insensitivity syndrome,Anemia Anemia Angiokeratoma Corporis Angiokeratoma Corporis 2023248067 10

Diffusum, Angiomatosis Diffusum, Angiomatosis retinae retinae (von (von Hippel-Lindau Hippel-Lindau disease) disease) ApertApert syndrome, syndrome, Arachnodactyly Arachnodactyly

(Marfan syndrome), (Marfan syndrome), Stickler Stickler syndrome, syndrome, Arthrochalasis Arthrochalasis multiplex multiplex congenital congenital (Ehlers-Danlos (Ehlers-Danlos

syndrome#arthrochalasia type) ataxia syndrome#arthrochalasia type) ataxia telangiectasia, telangiectasia, Rett Rettsyndrome, syndrome, primary pulmonary primary pulmonary

hypertension, Sandhoff hypertension, Sandhoff disease, disease, neurofibromatosis neurofibromatosis typeBeare-Stevenson type II, II, Beare-Stevenson cutis cutis gyrata gyrata syndrome, Mediterranean syndrome, Mediterranean fever, fever, familial, familial, Benjamin Benjamin syndrome, syndrome, beta-thalassemia, beta-thalassemia, Bilateral Bilateral

Acoustic Neurofibromatosis Acoustic Neurofibromatosis(neurofibromatosis (neurofibromatosis type type II),II), factorV V factor Leiden Leiden thrombophilia, thrombophilia, Bloch Bloch-

Sulzberger syndrome Sulzberger syndrome (incontinentiapigmenti), (incontinentia pigmenti),Bloom Bloom syndrome, syndrome, X-linked X-linked sideroblastic sideroblastic anemia, anemia,

Bonnevie-Ullrichsyndrome Bonnevie-Ullrich syndrome (Turner (Turner syndrome), syndrome), Bourneville Bourneville disease disease (tuberous (tuberous sclerosis), sclerosis), prionprion

disease, Birt-Hogg-Dub6 disease, Birt-Hogg-Dubé syndrome, syndrome, Brittle Brittle bone disease bone disease (osteogenesis (osteogenesis imperfecta), imperfecta), Broad Broad Thumb-Hallux syndrome Thumb-Hallux syndrome(Rubinstein-Taybi (Rubinstein-Taybisyndrome), syndrome),Bronze Bronze Diabetes/Bronzed Diabetes/Bronzed Cirrhosis Cirrhosis

(hemochromatosis), Bulbospinal (hemochromatosis), Bulbospinal muscular muscular atrophy atrophy (Kennedy's (Kennedy's disease), disease), Burger-Grutz Burger-Grutz syndrome syndrome

(lipoprotein lipase (lipoprotein lipase deficiency), deficiency), CGD Chronic CGD Chronic granulomatous granulomatous disorder, disorder, Campomelic Campomelic dysplasia, dysplasia,

biotinidase deficiency, biotinidase deficiency, Cardiomyopathy Cardiomyopathy (Noonan (Noonan syndrome), syndrome), Cri du Cri duCAVD chat, chat,(congenital CAVD (congenital absence of the absence of the vas vas deferens), deferens), Caylor Caylor cardiofacial cardiofacial syndrome (CBAVD), syndrome (CBAVD), CEPCEP (congenital (congenital

erythropoietic porphyria), cystic erythropoietic porphyria), cysticfibrosis, fibrosis,congenital congenital hypothyroidism, hypothyroidism, Chondrodystrophy Chondrodystrophy

syndrome (achondroplasia), otospondylomegaepiphyseal syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, dysplasia, Lesch-Nyhan syndrome, Lesch-Nyhan syndrome,

galactosemia, galactosemia, Ehlers-Danlos Ehlers-Danlos syndrome, Thanatophoric Thanatophoric dysplasia, dysplasia, Coffin-Lowry Coffin-Lowry syndrome, syndrome, Cockayne syndrome, Cockayne syndrome, (familial (familial adenomatous adenomatous polyposis), polyposis), Congenital Congenital erythropoietic erythropoietic porphyria, porphyria,

Congenital heart disease, Congenital heart disease, Methemoglobinemia/Congenital Methemoglobinemia/Congenital methaemoglobinaemia, methaemoglobinaemia, achondroplasia, X-linked X-linkedsideroblastic sideroblastic anemia, anemia,Connective Connective tissuedisease, tissue disease,Conotruncal Conotruncal anomaly anomaly

face syndrome, Cooley'sAnemia syndrome, Cooley's Anemia (beta-thalassemia), (beta-thalassemia), Copper Copper storage storage disease disease (Wilson's (Wilson's disease), disease),

Copper transport disease Copper transport disease (Menkes disease), hereditary (Menkes disease), hereditarycoproporphyria, coproporphyria,Cowden syndrome, Cowden syndrome,

Craniofacial dysarthrosis dysarthrosis (Crouzon (Crouzon syndrome), syndrome), Creutzfeldt-Jakob Creutzfeldt-Jakob disease disease (prion disease), (prion disease),

Cockayne syndrome,Cowden Cockayne syndrome, Cowden syndrome, syndrome, Curschmann-Batten-Steinert Curschmann-Batten-Steinert syndrome syndrome (myotonic (myotonic

dystrophy), Beare-Stevenson cutis dystrophy), Beare-Stevenson cutis gyrata gyratasyndrome, syndrome, primary primary hyperoxaluria, hyperoxaluria, spondyloepimetaphysealdysplasia spondyloepimetaphyseal dysplasia (Strudwick (Strudwick type), type), muscular muscular dystrophy, dystrophy, Duchenne Duchenne and Becker and Becker

types (DBMD), types (DBMD), Usher Usher syndrome, syndrome, Degenerative Degenerative nerve nerve diseases diseases including including de Grouchy de Grouchy syndrome syndrome

202 and Dejerine-Sottas syndrome, syndrome,developmental developmental 2023248067 10 Oct 2023 and Dejerine-Sottas disabilities,distal disabilities, distal spinal spinal muscular atrophy, type muscular atrophy, type V, V, insensitivity syndrome, androgen insensitivity androgen syndrome, Diffuse Globoid DiffuseGloboid Body Body Sclerosis Sclerosis (Krabbe (Krabbe disease), disease), Di George's Di George's syndrome, Dihydrotestosterone syndrome, Dihydrotestosterone receptor receptor deficiency, deficiency, androgen androgen insensitivity insensitivity syndrome, syndrome, Down Down syndrome, Dwarfism, syndrome, Dwarfism, erythropoietic erythropoietic protoporphyria protoporphyria Erythroid Erythroid 5-aminolevulinate 5-aminolevulinate synthetase synthetase deficiency, deficiency, Erythropoietic Erythropoietic porphyria, erythropoietic erythropoietic protoporphyria, protoporphyria, erythropoietic erythropoietic uroporphyria, uroporphyria,

Friedreich'sataxia,, Friedreich's ataxia,,familial familialparoxysmal paroxysmal polyserositis, polyserositis, porphyria porphyria cutanea cutanea tarda, tarda, pressure familial familial pressure sensitive sensitive neuropathy, neuropathy, primary primary pulmonary hypertension (PPH), pulmonary hypertension (PPH), Fibrocystic Fibrocystic disease disease of of the the pancreas, fragile pancreas, fragile XXsyndrome, syndrome, galactosemia, galactosemia, genetic genetic brain disorders, brain disorders, Giant Giant cell cell hepatitis hepatitis

(Neonatal hemochromatosis), Gronblad-Strandberg (Neonatal hemochromatosis), Gronblad-Strandberg syndrome syndrome(pseudoxanthoma (pseudoxanthoma elasticum), elasticum),

Gunther disease(congenital Gunther disease (congenitalerythropoietic erythropoieticporphyria), porphyria),haemochromatosis, haemochromatosis, Hallgren Hallgren syndrome, syndrome,

sickle cell anemia, sickle cell hemophilia,hepatoerythropoietic anemia, hemophilia, hepatoerythropoieticporphyria porphyria (HEP), (HEP), Hippel-Lindau Hippel-Lindau disease disease

(von Hippel-Lindau (von Hippel-Lindau disease), disease), Huntington's Huntington's disease, disease, Hutchinson-Gilford Hutchinson-Gilford progeriaprogeria syndrome syndrome

(progeria), (progeria),Hyperandrogenism, Hyperandrogenism, Hypochondroplasia, Hypochromicanemia, Hypochondroplasia, Hypochromic anemia,Immune Immune system system

disorders, including X-linked disorders, including X-linkedsevere severe combined combined immunodeficiency, immunodeficiency, Insley-Astley Insley-Astley syndrome,syndrome,

Kennedy's syndrome, Kennedy's syndrome, Jackson-Weiss Jackson-Weisssyndrome, syndrome,Joubert Joubertsyndrome, syndrome,Lesch-Nyhan Lesch-Nyhan syndrome, syndrome,

Jackson-Weiss syndrome, Jackson-Weiss syndrome, Kidney Kidney diseases, diseases, including including hyperoxaluria, hyperoxaluria, Klinefelter's Klinefelter's syndrome, syndrome,

Kniest dysplasia, Kniest dysplasia, Lacunar Lacunar dementia,Langer-Saldino dementia,Langer-Saldino achondrogenesis, achondrogenesis, ataxia telangiectasia, ataxia telangiectasia,

Lynchsyndrome, Lynch syndrome, Lysyl-hydroxylase Lysyl-hydroxylase deficiency, deficiency, Machado-Joseph Machado-Joseph disease, disease, Metabolic Metabolic disorders, disorders,

including Kniest Kniest dysplasia, dysplasia, Marfan Marfansyndrome, syndrome, Movement Movement disorders, disorders, Mowat-Wilson Mowat-Wilson syndrome,syndrome,

cystic cystic fibrosis, fibrosis,Muenke syndrome,Multiple Muenke syndrome, Multipleneurofibromatosis, neurofibromatosis, Nance-Insley Nance-Insley syndrome, syndrome, Nance Nance-

Sweeney chondrodysplasia, Sweeney chondrodysplasia, Niemann-Pick Niemann-Pickdisease, disease,Noack Noack syndrome syndrome (Pfeiffer (Pfeiffer syndrome), syndrome),

Osler-Weber-Rendu disease, Osler-Weber-Rendu disease, Peutz-Jeghers Peutz-Jeghers syndrome, syndrome, Polycystic Polycystic kidney kidney disease, disease, polyostotic polyostotic

fibrous fibrous dysplasia (McCune-Albright (McCune-Albright syndrome), syndrome), Peutz-Jeghers Peutz-Jeghers syndrome, syndrome, Prader-Labhart-Willi Prader-Labhart-Willi

syndrome, hemochromatosis, primary syndrome, hemochromatosis, primaryhyperuricemia hyperuricemiasyndrome syndrome (Lesch-Nyhan (Lesch-Nyhan syndrome), syndrome),

primary pulmonary primary pulmonary hypertension, hypertension, primary primary senile senile degenerative degenerative dementia, dementia, prionprion disease, disease, progeria progeria

(Hutchinson GilfordProgeria (Hutchinson Gilford ProgeriaSyndrome), Syndrome), progressive progressive chorea, chorea, chronic chronic hereditary hereditary (Huntington) (Huntington)

(Huntington's disease), (Huntington's disease), progressive progressivemuscular muscular atrophy, atrophy, spinalspinal muscular muscular atrophy,atrophy, propionicpropionic

acidemia, protoporphyria, acidemia, protoporphyria, proximal proximalmyotonic myotonic dystrophy, dystrophy, pulmonary pulmonary arterial arterial hypertension, hypertension, PXE PXE (pseudoxanthoma elasticum), (pseudoxanthoma elasticum), Rb Rb (retinoblastoma), (retinoblastoma), Recklinghausen Recklinghausen disease disease (neurofibromatosis (neurofibromatosis

type I), type I), Recurrent Recurrent polyserositis, polyserositis,Retinal Retinaldisorders, disorders,Retinoblastoma, Retinoblastoma, Rett Rett syndrome, RFALS syndrome, RFALS type type

3, 3, Ricker syndrome,Riley-Day Ricker syndrome, Riley-Day syndrome, syndrome, Roussy-Levy Roussy-Levy syndrome, syndrome, severe achondroplasia severe achondroplasia with with

203 developmental delay delay and and acanthosis acanthosis nigricans 2023248067 10 Oct 2023 developmental nigricans(SADDAN), Li-Fraumeni syndrome, (SADDAN), Li-Fraumeni syndrome,sarcoma, sarcoma, breast, leukemia, breast, and adrenal leukemia, and adrenal gland gland(SBLA) (SBLA) syndrome, syndrome, sclerosis sclerosis tuberose tuberose (tuberous (tuberous sclerosis), sclerosis),

SDAT, SED SDAT, SED congenital(spondyloepiphyseal congenital (spondyloepiphyseal dysplasia dysplasia congenita), congenita), SED SEDStrudwick Strudwick (spondyloepimetaphyseal dysplasia, (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc Strudwick type), (spondyloepiphyseal dysplasia SEDc (spondyloepiphyseal dysplasia congenita) SEMD,Strudwick congenita) SEMD, Strudwick typetype (spondyloepimetaphyseal (spondyloepimetaphyseal dysplasia, dysplasia, Strudwick Strudwick type), type),

Shprintzen syndrome, Shprintzen syndrome, Skin Skin pigmentation pigmentationdisorders, disorders, Smith-Lemli-Opitz Smith-Lemli-Opitz syndrome, syndrome,South- South African genetic African genetic porphyria porphyria(variegate (variegate porphyria), porphyria), infantile-onset infantile-onset ascending ascending hereditary hereditary spastic spastic

paralysis, Speech paralysis, andcommunication Speech and communication disorders, disorders, sphingolipidosis,Tay-Sachs sphingolipidosis, Tay-Sachs disease, disease,

spinocerebellar spinocerebellar ataxia, Stickler syndrome, ataxia, Stickler syndrome, stroke, stroke, androgen androgen insensitivitysyndrome, insensitivity syndrome, tetrahydrobiopterin deficiency, tetrahydrobiopterin deficiency, beta-thalassemia, beta-thalassemia,Thyroid Thyroid disease, disease, Tomaculous Tomaculous neuropathy neuropathy

(hereditary (hereditary neuropathy withliability neuropathy with liability to to pressure pressure palsies), palsies), Treacher Treacher Collins Collins syndrome, TriploX X syndrome, Triplo

syndrome syndrome ( (triple triple X syndrome),Trisomy X syndrome), Trisomy21 21 (Down (Down syndrome), syndrome), Trisomy Trisomy X, VHL X, VHL syndrome syndrome (von (von Hippel-Lindaudisease), Hippel-Lindau disease),Vision Visionimpairment impairment and and blindness blindness (Alstr6m (Alström syndrome), syndrome), VrolikVrolik disease, disease,

Waardenburg syndrome, Waardenburg syndrome, Warburg Warburg Sjo Fledelius Sjo Fledelius Syndrome, Syndrome, Weissenbacher-Zweymiiller Weissenbacher-Zweymüller

syndrome, Wolf-Hirschhorn syndrome, syndrome, Wolf-Hirschhorn syndrome,Wolff Wolff Periodicdisease, Periodic disease, Weissenbacher-Zweymiller Weissenbacher-Zweymiiller syndrome and Xeroderma syndrome and Xeroderma pigmentosum, pigmentosum, among amongothers. others.

[0440] The term

[0440] The term "neoplasia" "neoplasia" or "cancer" or "cancer" is used used throughout is throughout the specification the specification to refer to refer to theto the pathological process pathological process that thatresults results ininthe theformation formation andand growth growth of a of a cancerous cancerous or malignant or malignant

neoplasm, i.e., neoplasm, i.e., abnormal abnormaltissue tissuethat thatgrows grows by by cellular cellular proliferation,often proliferation, often more more rapidly rapidly thanthan

normal and normal andcontinues continuestotogrow grow after after thethe stimuli stimuli thatinitiated that initiated the the new newgrowth growth cease. cease. Malignant Malignant

neoplasmsshow neoplasms show partialororcomplete partial complete lack lack of of structuralorganization structural organizationandand functional functional coordination coordination

with the with the normal tissue and normal tissue and most mostinvade invadesurrounding surrounding tissues,metastasize tissues, metastasizetotoseveral severalsites, sites, and are and are

likely likely to to recur recur after afterattempted attempted removal andtotocause removal and causethe thedeath deathofofthe thepatient patientunless unlessadequately adequately treated. As treated. As used herein, the used herein, the term term neoplasia neoplasia isis used used toto describe describe all all cancerous cancerous disease diseasestates states and and embracesororencompasses embraces encompasses the the pathological pathological process process associated associated with malignant with malignant hematogenous, hematogenous,

ascitic and ascitic and solid solid tumors. Exemplarycancers tumors. Exemplary cancers which which may may be treated be treated bypresent by the the present compounds compounds

either either alone alone or or in in combination withatat least combination with least one additional anti-cancer one additional anti-cancer agent agent include include squamous- squamous cell cell carcinoma, basal cell carcinoma, basal cell carcinoma, adenocarcinoma,hepatocellular carcinoma, adenocarcinoma, hepatocellularcarcinomas, carcinomas, andand renal renal cell cell

carcinomas, cancerofofthe carcinomas, cancer thebladder, bladder, bowel, bowel,breast, breast, cervix, cervix, colon, colon, esophagus, esophagus,head, head,kidney, kidney,liver, liver, lung, lung, neck, ovary, ovary, pancreas, pancreas, prostate, prostate,and and stomach; stomach; leukemias; benign and leukemias; benign andmalignant malignantlymphomas, lymphomas, particularly Burkitt's particularly Burkitt'slymphoma and Non-Hodgkin's lymphoma and Non-Hodgkin'slymphoma; lymphoma; benign benign and malignant and malignant

204 melanomas; myeloproliferative myeloproliferative diseases; 2023248067 10 Oct 2023 melanomas; diseases; sarcomas, including sarcomas, including Ewing's Ewing's sarcoma, sarcoma, hemangiosarcoma, hemangiosarcoma, Kaposi's Kaposi's sarcoma, sarcoma, liposarcoma, liposarcoma, myosarcomas, myosarcomas, peripheral peripheral neuroepithelioma, neuroepithelioma, synovial sarcoma, sarcoma,gliomas, gliomas, astrocytomas, astrocytomas, oligodendrogliomas, oligodendrogliomas, ependymomas, ependymomas, gliobastomas, gliobastomas, neuroblastomas, ganglioneuromas, neuroblastomas, ganglioneuromas, gangliogliomas, gangliogliomas, medulloblastomas, medulloblastomas, pineal pineal cell cell tumors, tumors, meningiomas, meningeal meningiomas, meningeal sarcomas, sarcomas, neurofibromas, neurofibromas, and and Schwannomas; Schwannomas;bowel bowel cancer,breast cancer, breast cancer, prostate cancer, cancer, prostate cancer, cervical cervical cancer, cancer, uterine uterine cancer, cancer, lung cancer, ovarian lung cancer, ovarian cancer, cancer, testicular testicular cancer, cancer, thyroid cancer, cancer, astrocytoma, esophagealcancer, astrocytoma, esophageal cancer, pancreatic pancreatic cancer, cancer, stomach stomachcancer, cancer,liver liver cancer, cancer, colon cancer, melanoma; colon cancer, melanoma; carcinosarcoma, carcinosarcoma, Hodgkin's Hodgkin'sdisease, disease, Wilms' Wilms'tumor tumor and and teratocarcinomas. Additional teratocarcinomas. Additionalcancers cancers which which may may be treated be treated using using compounds compounds according according to the to the present disclosure present disclosure include, include, for for example, T-lineage Acute example, T-lineage Acutelymphoblastic lymphoblasticLeukemia Leukemia (T-ALL), (T-ALL), T- T lineage lymphoblastic lymphoblastic Lymphoma Lymphoma (T-LL), (T-LL), Peripheral Peripheral T-cell T-cell lymphoma, lymphoma, Adult T-cell Adult T-cell Leukemia, Leukemia,

Pre-B ALL, Pre-B ALL, Pre-B Pre-BLymphomas, Lymphomas, Large Large B-cell B-cell Lymphoma, Lymphoma, Burkitts Burkitts Lymphoma, Lymphoma, B-cellB-cell ALL, ALL,

Philadelphia chromosome Philadelphia chromosome positive positive ALLALL and and Philadelphia Philadelphia chromosome chromosome positive positive CML. CML.

[0441] TheThe

[0441] termterm "bioactive "bioactive agent" agent" is isused usedtotodescribe describe ananagent, agent, other other than than aa compound compound

according to according to the the present present disclosure, disclosure, which which is is used used in in combination with the combination with the present present compounds compounds as as

an agent agent with withbiological biologicalactivity activity totoassist assist in in effecting effecting an an intended intendedtherapy, therapy,inhibition inhibitionand/or and/or prevention/prophylaxis for prevention/prophylaxis forwhich whichthethepresent presentcompounds compounds are used. are used. Preferred Preferred bioactive bioactive agentsagents

for for use use herein herein include include those those agents agents which have pharmacological which have pharmacologicalactivity activitysimilar similar to to that that for forwhich which

the present the compounds present compounds areare used used or or administered administered and include and include for example, for example, anti-cancer anti-cancer agents,agents,

antiviral agents, antiviral agents,especially especiallyincluding includinganti-HIV anti-HIV agents agents and and anti-HCV agents, antimicrobial anti-HCV agents, antimicrobial agents, agents, antifungalagents, antifungal agents,etc. etc.

[0442] The term

[0442] The "additional term "additional anti-cancer anti-cancer agent" agent" is used istoused to describe describe an anti-cancer an anti-cancer agent, agent, which may which maybe be combined combined with with compounds compounds according according to the disclosure to the present present disclosure to treat to treat cancer. cancer. These agents include, These agents include, for for example, example,everolimus, everolimus,trabectedin, trabectedin,abraxane, abraxane,TLKTLK 286,286, AV-299, AV-299, DN- DN 101, pazopanib, 101, pazopanib,GSK690693, RTA744, GSK690693, RTA 744,ONON0910.Na, 0910.Na,AZD AZD 6244 6244 (ARRY-142886), (ARRY-142886), AMN-107, AMN-107,

TKI-258, GSK461364, TKI-258, GSK461364,AZD AZD 1152, 1152, enzastaurin,vandetanib, enzastaurin, vandetanib, ARQ-197, ARQ-197,MK-0457, MK-0457, MLN8054, MLN8054,

PHA-739358, PHA-739358, R-763, R-763, AT-9263, AT-9263, a FLT-3 a FLT-3 inhibitor, inhibitor, a VEGFR a VEGFR inhibitor, inhibitor, an EGFR an TK EGFR TK inhibitor, inhibitor, an aurora aurora kinase kinase inhibitor, inhibitor, a PIK-1 modulator,a aBcl-2 PIK-1 modulator, Bcl-2inhibitor, inhibitor,ananHDAC HDAC inhbitor, inhbitor, a c-MET a c-MET

inhibitor, inhibitor, aa PARP inhibitor, aa Cdk PARP inhibitor, Cdkinhibitor, inhibitor, an an EGFR EGFR TK TK inhibitor, inhibitor, an an IGFR-TK IGFR-TK inhibitor, inhibitor, an an anti-HGF antibody, anti-HGF antibody, aa PI3 P13 kinase kinase inhibitor, inhibitor, an an AKT inhibitor, an AKT inhibitor, mTORCI/2 an mTORC1/2 inhibitor, a inhibitor, a JAK/STAT JAK/STAT inhibitor, inhibitor, a checkpoint-i a checkpoint-1 or 2 or 2 inhibitor, inhibitor, a focal a focal adhesion adhesion kinase kinase inhibitor, inhibitor, a Mapa Map

205 kinase kinase kinase (mek) (mek) inhibitor, inhibitor, a VEGF trapantibody, antibody,pemetrexed, pemetrexed, erlotinib,dasatanib, 2023248067 10 Oct 2023 kinase VEGF trap erlotinib, dasatanib,nilotinib, nilotinib, decatanib, panitumumab, decatanib, panitumumab, amrubicin, amrubicin, oregovomab, oregovomab, Lep-etu, Lep-etu, nolatrexed, nolatrexed, azd2171, azd2171, batabulin, batabulin, ofatumumab, ofatumumab, zanolimumab,edotecarin, zanolimumab, edotecarin,tetrandrine, tetrandrine,rubitecan, rubitecan,tesmilifene, tesmilifene, oblimersen, oblimersen, ticilimumab, ticilimumab, ipilimumab, ipilimumab, gossypol, gossypol,Bio Bio 111, 111, 131-I-TM-601, 131-I-TM-601, ALT-110, BIO140, ALT-110, BIO 140,CCCC 8490, 8490, cilengitide, cilengitide, gimatecan, IL13-PE38QQR, gimatecan, IL13-PE38QQR, INO 1001, IPdR INO 1001, KRX-0402,lucanthone, IPdR1 KRX-0402, lucanthone,LY317615, LY317615, neuradiab, vitespan, neuradiab, vitespan, Rta 744, Sdx Rta 744, Sdx 102, 102, talampanel, talampanel, atrasentan, atrasentan, Xr 311, romidepsin, Xr 311, romidepsin, ADS-100380, ADS-100380, sunitinib, 5-fluorouracil,vorinostat, sunitinib, 5-fluorouracil, vorinostat, etoposide, etoposide, gemcitabine, gemcitabine, doxorubicin, doxorubicin, liposomal liposomal doxorubicin,doxorubicin,

5'-deoxy-5-fluorouridine, vincristine, temozolomide, 5'-deoxy-5-fluorouridine, vincristine, ZK-304709, temozolomide, ZK-304709, seliciclib;PD0325901, seliciclib; PD0325901, AZD- AZD

6244, capecitabine, L-Glutamic 6244, capecitabine, L-Glutamic acid,acid, N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H- N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H pyrrolo[2,3 pyrrolo[2,3-

d]pyrimidin-5-yl)ethyl]benzoyl]-, disodium d]pyrimidin-5-yl)ethyl]benzoy1]-, disodium salt, salt, heptahydrate, heptahydrate, camptothecin, camptothecin, PEG-labeled PEG-labeled

irinotecan, tamoxifen, irinotecan, tamoxifen, toremifene toremifene citrate, citrate, anastrazole, anastrazole, exemestane, exemestane, letrozole, letrozole,

DES(diethylstilbestrol), estradiol, estrogen, DES(diethylstilbestrol), estradiol, estrogen, conjugated conjugatedestrogen, estrogen,bevacizumab, bevacizumab, IMC-1CI1, IMC-1C11,

CHIR-258); 3-[5-(methylsulfonylpiperadinemethyl)- CHIR-258); 3-[5-(methylsulfonylpiperadinemethyl)- indolyl-quinolone, indolyl-quinolone, vatalanib, vatalanib, AG-013736, AG-013736,

AVE-0005,goserelin AVE-0005, goserelin acetate, acetate, leuprolide leuprolide acetate, acetate, triptorelin triptorelin pamoate, pamoate, medroxyprogesterone medroxyprogesterone

acetate, hydroxyprogesterone acetate, caproate,megestrol hydroxyprogesterone caproate, megestrol acetate,raloxifene, acetate, raloxifene,bicalutamide, bicalutamide, flutamide, flutamide,

nilutamide, megestrol nilutamide, megestrol acetate, acetate, CP-724714; CP-724714;TAK-165, TAK-165, HKI-272, HKI-272, erlotinib, erlotinib, lapatanib, lapatanib, canertinib, canertinib,

ABX-EGF ABX-EGF antibody,erbitux, antibody, erbitux,EKB-569, EKB-569, PKI-166, PKI-166, GW-572016, GW-572016, Ionafarnib, Ionafarnib, BMS-214662, BMS-214662,

tipifarnib; tipifarnib;amifostine, amifostine,NVP-LAQ824, NVP-LAQ824, suberoylanalide suberoyl analidehydroxamic hydroxamic acid, acid, valproic valproic acid, acid,

trichostatin A,A, FK-228, trichostatin SU11248,sorafenib, FK-228, SU11248, sorafenib, KRN951 KRN951 , aminoglutethimide, , aminoglutethimide, arnsacrine, arnsacrine,

anagrelide, L-asparaginase, Bacillus anagrelide, L-asparaginase, Bacillus Calmette-Guerin Calmette-Guerin (BCG) (BCG) vaccine, vaccine, adriamycin, adriamycin, bleomycin, bleomycin,

buserelin, busulfan, buserelin, busulfan, carboplatin, carboplatin, carmustine, carmustine,chlorambucil, chlorambucil, cisplatin, cisplatin, cladribine,clodronate, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, cyproterone, cytarabine, dacarbazine, dactinomycin, dactinomycin,daunorubicin, daunorubicin,diethylstilbestrol, diethylstilbestrol, epirubicin, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, fludarabine, fludrocortisone, fluoxymesterone,flutamide, flutamide,gleevec, gleevec,gemcitabine, gemcitabine, hydroxyurea, hydroxyurea,

idarubicin, idarubicin, ifosfamide, ifosfamide, imatinib, imatinib,leuprolide, leuprolide,levamisole, levamisole,lomustine, lomustine,mechlorethamine, melphalan, mechlorethamine, melphalan,

6-mercaptopurine, mesna, 6-mercaptopurine, mesna, methotrexate, methotrexate, mitomycin, mitomycin, mitotane, mitotane,mitoxantrone, mitoxantrone,nilutamide, nilutamide, octreotide, oxaliplatin,pamidronate, octreotide, oxaliplatin, pamidronate, pentostatin, pentostatin, plicamycin, plicamycin, porfimer, porfimer, procarbazine, procarbazine, raltitrexed,raltitrexed,

rituximab, streptozocin, rituximab, streptozocin, teniposide, teniposide, testosterone, testosterone, thalidomide, thalidomide, thioguanine, thioguanine, thiotepa, thiotepa, tretinoin, tretinoin, vindesine, 13-cis-retinoic vindesine, 13-cis-retinoic acid, acid, phenylalanine phenylalanine mustard, mustard, uracil mustard, uracil mustard, estramustine, estramustine, altretamine,altretamine,

floxuridine, floxuridine, 5-deooxyuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, deoxycoformycin, calcitriol, calcitriol,valrubicin, valrubicin, mithramycin, vinblastine, vinorelbine, mithramycin, vinblastine, vinorelbine, topotecan, topotecan,razoxin, razoxin,marimastat, marimastat, COL-3, neovastat, BMS-275291 COL-3, neovastat, , squalamine,endostatin, BMS-275291, squalamine, endostatin, SU5416, SU5416,SU6668, SU6668, EMD121974, EMD121974,

206 interleukin-12, IM862, angiostatin, 2023248067 10 Oct 2023 interleukin-12, IM862, angiostatin, vitaxin, vitaxin, droloxifene, droloxifene, idoxyfene, idoxyfene, spironolactone, spironolactone, finasteride, finasteride, cimitidine, trastuzumab, denileukin cimitidine, trastuzumab, diftitox,gefitinib, bortezimib, denileukindiftitox,gefitinib, bortezimib,paclitaxel, cremophor-free paclitaxel, cremophor-free paclitaxel, docetaxel, paclitaxel, epithilone docetaxel, B, B, epithilone BMS- 247550, BMS-310705, BMS- 247550, BMS-310705, droloxifene, droloxifene, 4-4 hydroxytamoxifen,pipendoxifene, hydroxytamoxifen, pipendoxifene, ERA-923, ERA-923, arzoxifene, arzoxifene, fulvestrant, fulvestrant, acolbifene, acolbifene, lasofoxifene, lasofoxifene, idoxifene, idoxifene,TSE-424, TSE-424, HMR- 3339,ZK186619, HMR- 3339, ZK186619, topotecan,PTK787/ZK topotecan, PTK787/ZK 222584, 222584, VX-745, VX-745, PD PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, 184352, rapamycin, 40-0-(2-hydroxyethyl)-rapamycin, temsirolimus, temsirolimus, AP-23573, AP-23573, RAD001, RADOO1, ABT-578, BC-210,LY294002, ABT-578, BC-210, LY294002, LY292223, LY292223, LY292696, LY292696, LY293684, LY293684, LY293646, LY293646, wortmannin, wortmannin,

ZM336372,L-779,450, ZM336372, L-779,450,PEG-filgrastim, PEG-filgrastim,darbepoetin, darbepoetin,erythropoietin, erythropoietin, granulocyte granulocyte colony- colony stimulating factor, factor, zolendronate, zolendronate,prednisone, prednisone, cetuximab, cetuximab, granulocyte granulocyte macrophage macrophage colony- colony stimulating factor,histrelin, stimulating factor, histrelin,pegylated pegylated interferon interferon alfa-2a, alfa-2a, interferon interferon alfa-2a, alfa-2a, pegylated pegylated interferon interferon

alfa-2b, interferon alfa-2b, interferon alfa-2b, alfa-2b,azacitidine, azacitidine,PEG-L-asparaginase, PEG-L-asparaginase, lenalidomide, lenalidomide, gemtuzumab, gemtuzumab,

hydrocortisone, interleukin-11, hydrocortisone, interleukin-11, dexrazoxane, dexrazoxane, alemtuzumab, alemtuzumab, all-transretinoicacid, all-transretinoic acid,ketoconazole, ketoconazole, interleukin-2, interleukin-2, megestrol, immuneglobulin, megestrol, immune globulin, nitrogen nitrogen mustard, mustard, methylprednisolone, methylprednisolone,

ibritgumomab tiuxetan,androgens, ibritgumomab tiuxetan, androgens,decitabine, decitabine,hexamethylmelamine, hexamethylmelamine, bexarotene, bexarotene, tositumomab, tositumomab,

arsenic trioxide, arsenic trioxide, cortisone, cortisone,editronate, editronate,mitotane, mitotane,cyclosporine, cyclosporine,liposomal liposomal daunorubicin, daunorubicin, Edwina Edwina-

asparaginase, strontium asparaginase, strontium 89, 89, casopitant, casopitant, netupitant, netupitant, an an NK-1 NK-1receptor receptor antagonist, antagonist, palonosetron, palonosetron,

aprepitant, diphenhydramine, aprepitant, hydroxyzine,metoclopramide, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, lorazepam, alprazolam, alprazolam, haloperidol, haloperidol,

droperidol, dronabinol, droperidol, dronabinol, dexamethasone, dexamethasone, methylprednisolone, methylprednisolone, prochlorperazine, prochlorperazine, granisetron, granisetron,

ondansetron, dolasetron, ondansetron, dolasetron, tropisetron, tropisetron, pegfilgrastim, pegfilgrastim, erythropoietin, erythropoietin, epoetin epoetin alfa, alfa, darbepoetin darbepoetin alfa alfa and mixtures and mixtures thereof. thereof.

[0443] TheThe

[0443] termterm "anti-HIV "anti-HIV agent" agent" or "additionalanti-HIV or "additional anti-HIVagent" agent"includes, includes, for for example, example, nucleoside reverse nucleoside reverse transcriptase transcriptase inhibitors inhibitors (NRTI), other non-nucloeoside (NRTI), other non-nucloeosidereverse reverse transcriptase transcriptase

inhibitors (i.e., those inhibitors (i.e., which those which areare notnot representative representative of theofpresent the present disclosure), disclosure), proteaseprotease inhibitors, inhibitors, fusion inhibitors, inhibitors, among others, exemplary among others, compounds exemplary compounds of which of which may may include, include, for example, for example, 3TC 3TC

(Lamivudine), AZT (Lamivudine), AZT (Zidovudine), (Zidovudine), (-)-FTC, (-)-FTC, ddI ddl (Didanosine), (Didanosine), ddC ddC (zalcitabine), (zalcitabine), abacavir abacavir (ABC), (ABC),

tenofovir (PMPA), tenofovir D-D4FC (PMPA), D-D4FC (Reverset),D4T (Reverset), D4T (Stavudine),Racivir, (Stavudine), Racivir, L-FddC, L-FddC,L-FD4C, L-FD4C, NVPNVP

(Nevirapine), (Nevirapine), DLV (Delavirdine), EFV DLV (Delavirdine), EFV(Efavirenz), (Efavirenz), SQVM SQVM (Saquinavir (Saquinavir mesylate), mesylate), RTV RTV

(Ritonavir), (Ritonavir), IDV (Indinavir), SQV IDV (Indinavir), SQV (Saquinavir), (Saquinavir), NFVNFV (Nelfinavir), (Nelfinavir), APV (Amprenavir), APV (Amprenavir), LPV LPV (Lopinavir), fusion inhibitors (Lopinavir), inhibitorssuch such as as T20, T20, among others, fuseon among others, fuseon and andmixtures mixturesthereof, thereof, including including anti-HIV compounds anti-HIV compounds presently presently in in clinicaltrials clinical trials or in in development. development.

207

Oct 2023 Other

[0444] Other

[0444] anti-HIV anti-HIV agents agents which may may which be used be used in coadministration withwith in coadministration compounds compounds

according to the according to the present presentdisclosure disclosureinclude, include,for forexample, example,other other NNRTI's NNRTI's (i.e., (i.e., other other thanthan the the

NNRTI's NNRTI's according according to the to the present present disclosure) disclosure) may may be selected be selected from from the the consisting group group consisting of of nevirapine (BI-R6-587), nevirapine delavirdine (U-90152S/T), (BI-R6-587), delavirdine efavirenz (DMP-266), (U-90152S/T), efavirenz UC-781(N-[4- (DMP-266), UC-781 (N-[4 2023248067 10

chloro-3-(3-methyl-2-butenyloxy)phenyl]-2methyl3-furancarbothiamide), etravirine chloro-3-(3-methyl-2-butenyloxy)phenyl]-2methyl3-furancarbothiamide), etravirine (TMC125), (TMC125),

Trovirdine (Ly300046.HCI), Trovirdine (Ly300046.HCl),MKC-442 MKC-442 (emivirine, (emivirine, coactinon), coactinon), HI-236, HI-236, HI-240, HI-240, HI-280, HI-280, HI-281,HI-281,

rilpivirine rilpivirine (TMC-278), (TMC-278),MSC-127, MSC-127, HBY 097, DMP266, HBY 097, DMP266,Baicalin Baicalin(TJN-151) (TJN-151)ADAM-II ADAM-II (Methyl (Methyl

3',3'-dichloro-4',4"-dimethoxy-5',5"-bis(methoxycarbonyl)-6,6-diphenylhexenoate), ,3'-dichloro-4',4'-dimethoxy-5',5"-bis(methoxycarbonyl)-6,6-diphenylhexenoate) Methyl Methyl 3- 3 Bromo-5-(1-5-bromo-4-methoxy-3-(methoxycarbonyl)phenyl)hept-1-enyl)-2-methoxybenzoate Bromo-5-(1-5-bromo-4-methoxy-3-(methoxycarbonyl)phenyl)hept-l-enyl)-2-methoxybenzoate

(Alkenyldiarylmethane (Alkenyldiarylmethane analog, Adam analog, Adam analog), analog), (5-chloro-3-(phenylsulfinyl)-2' (5-chloro-3-(phenylsulfinyl)-2' indolecarboxamide), indolecarboxamide),AAP-BHAP (U-104489or orPNU-104489), AAP-BHAP (U-104489 PNU-104489),Capravirine Capravirine(AG-1549, (AG-1549,S-1153), S-1153), atevirdine (U-87201E),aurin atevirdine (U-87201E), aurintricarboxylic tricarboxylicacid acid(SD-095345), (SD-095345), 1-[(6-cyano-2-indolyl)carbonyl] 1-[(6-cyano-2-indolyl)carbonyl]-

4-[3-(isopropylamino)-2-pyridinyl]piperazine, 4-[3-(isopropylamino)-2-pyridinyl]piperazine, 1-[5-[[N-(methyl)methylsulfonylamino]-2 1-[5-[[N-(methyl)methylsulfonylamino]-2-

indolylcarbonyl-4-[3-(isopropylamino)-2-pyridinyl]piperazine, indolylcarbonyl-4-[3-(isopropylamino)-2-pyridinyl]piperazine, 1-[3-(Ethylamino)-2-[pyridinyl] 1-[3-(Ethylamino)-2-[pyridinyl]-

4-[(5-hydroxy-2-indolyl)carbonyl]piperazine, 4-[(5-hydroxy-2-indolyl)carbonyl]piperazine, 1-[(6-Formyl-2-indolyl)carbonyl]-4-[3 1-[(6-Formyl-2-indolyl)carbonyl]-4-[3-

(isopropylamino)-2-pyridinyl]piperazine, (isopropylamino)-2-pyridinyI]piperazine, 1-[[5-(Methylsulfonyloxy)-2-indoyly)carbonyl]-4-[3 1-[[5-(Methylsulfonyloxy)-2-indoyly)carbonyl]-4-[3-

(isopropylamino)-2-pyridinyl]piperazine, U88204E, U88204E, Bis(2-nitrophenyl)sulfone Bis(2-nitrophenyl)sulfone (NSC 633001), (NSC 633001),

Calanolide Calanolide AA(NSC675451), (NSC675451), Calanolide Calanolide B, 6-Benzyl-5-methyl-2-(cyclohexyloxy)pyrimidin-4 B, 6-Benzyl-5-methyl-2-(cyclohexyloxy)pyrinmidin-4-

one (DABO-546),DPC one (DABO-546), DPC 961,961, E-EBU, E-EBU, E-EBU-dm, E-EBU-dm, E-EPSeU, E-EPSeU, E-EPU, Foscarnet E-EPU, Foscarnet (Foscavir), (Foscavir),

HEPT HEPT (1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)thymine), (1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)thymine), HEPT-M HEPT-M (1-[(2 (1-[(2-

Hydroxyethoxy)methyl]-6-(3-methylphenyl)thio)thymine), Hydroxyethoxy)methyl]-6-(3-methylphenyl)thio)thymine), HEPT-S HEPT-S (1-[(2 (1-[(2-

Hydroxyethoxy)methyl]-6-(phenylthio)-2-thiothymine), Hydroxyethoxy)methyl]-6-(phenylthio)-2-thiothymine), Inophyllum InophyllumP,P, L-737,126, L-737,126, Michellamine Michellamine AA(NSC650898), (NSC650898), Michellamine Michellamine B (NSC649324), B (NSC649324), Michellamine Michellamine F, 6-(3,5 F, 6-(3,5-

Dimethylbenzyl)-1-[(2-hydroxyethoxy)methyl]-5-isopropyluracil, 6-(3,5-Dimethylbenzyl)-1 Dimethylbenzyl)-1-[(2-hydroxyethoxy)methyl]-5-isopropyluracil, 6-(3,5-Dimethylbenzyl)-1-

(ethyoxymethyl)-5-isopropyluracil, NPPS, (ethyoxymethyl)-5-isopropyluracil, NPPS, E-BPTU (NSC E-BPTU (NSC 648400), 648400), Oltipraz Oltipraz (4-Methyl-5 (4-Methyl-5-

(pyrazinyl)-3H-1,2-dithiole-3-thione), (pyrazinyl)-3H-1,2-dithiole-3-thione), N-{2-(2-Chloro-6-fluorophenethyl]-N'-(2 N-{2-(2-Chloro-6-fluorophenethyl]-N'-(2-

thiazolyl)thiourea thiazolyl)thiourea (PETT (PETT Cl,Cl, F derivative), F derivative), N-{2-(2,6-Difluorophenethyl]-N'-[2-(5 N-{2-(2,6-Difluorophenethy1]-N'-[2-(5-

bromopyridyl)]thiourea {PETT bromopyridyl)]thiourea {PETT derivative), derivative), N-{2-(2,6-Difluorophenethyl]-N'-[2-(5 N-{2-(2,6-Difluorophenethyl]-N'-[2-(5-

methylpyridyl)]thiourea {PETT methylpyridyl)]thiourea {PETT Pyridyl Pyridyl derivative), derivative), N-[2-(3-Fluorofuranyl)ethyl]-N'-[2-(5 N-[2-(3-Fluorofuranyl)ethyl]-N'-[2-(5-

chloropyridyl)]thiourea, chloropyridy1)]thiourea, N-[2-(2-Fluoro-6-ethoxyphenethyl)]-N'-[2-(5-bromopyridyl)]thiourea, N-[2-(2-Fluoro-6-ethoxyphenethyl)]-N'-[2-(5-bromopyridyl)]thiourea,

N-(2-Phenethyl)-N'-(2-thiazolyl)thiourea (LY-73497), N-(2-Phenethyl)-N'-(2-thiazolyl)thiourea (LY-73497), L-697,639, L-697,639, L-697,593, L-697,593, L-697,661, L-697,661, 3-[2- 3-[2

208

Oct 2023 (4,7-Difluorobenzoxazol-2-yl)ethyl}-5-ethyl-6-methyl(pypridin-2(1H)-thione(2-Pyridinone (4,7-Difluorobenzoxazol-2-yl)ethyl}-5-ethyl-6-methyl(pypridin-2(1H)-thione (2-Pyridinone Derivative), B-[[(2-Methoxy-5,6-dimethyl-3-pyridyl)methylJamine]-5-ethyl-6-methyl(pypridin- Derivative), 3-[[(2-Methoxy-5,6-dimethyl-3-pyridyl)methyl]amine]-5-ethyl-6-methyl(pypridin 2(1H)-thione, R82150, 2(1H)-thione, R82150,R82913, R82913, R87232, R87232, R88703, R88703, R89439 R89439 (Loviride), (Loviride), R90385,R90385, S-2720, S-2720, Suramin Suramin Sodium,TBZ Sodium, TBZ (Thiazolobenzimidazole, (Thiazolobenzimidazole, NSC 625487), NSC 625487), Thiazoloisoindol-5-one, Thiazoloisoindol-5-one, (+)(R)-9b-(3,5 (+)(R)-9b-(3,5- 2023248067 10

Dimethylphenyl-2,3-dihydrothiazolo[2,3-a]isoindol-5(9bH)-one, Dimethylphenyl-2,3-dihydrothiazolo[2,3-a]isoindol-5(9bH)-one, Tivirapine Tivirapine (R86183), (R86183), UC-38 UC-38 and UC-84,among and UC-84, among others. others.

[0445] The term

[0445] The term "pharmaceutically "pharmaceutically acceptable acceptable salt" is used throughout usedisthroughout salt" the specification the specification to to describe, where describe, applicable, aa salt where applicable, saltform form of of one one or or more of the more of the compounds described compounds described herein herein which which

are presented to are presented to increase increase the thesolubility solubility ofofthe the compound compound in the in the gastic gastic juices juices of the of the patient's patient's

gastrointestinal gastrointestinal tract tractininorder ordertotopromote promote dissolution dissolution and and the the bioavailability bioavailability of ofthe thecompounds. compounds.

Pharmaceutically acceptable Pharmaceutically acceptablesalts salts include include those those derived derived from pharmaceutically from pharmaceutically acceptable acceptable

inorganic or or organic bases bases and and acids, acids, where applicable. Suitable where applicable. Suitable salts salts include include those those derived derived from from

alkali alkali metals metals such as potassium such as andsodium, potassium and sodium,alkaline alkalineearth earthmetals metalssuch such as as calcium, calcium, magnesium magnesium

and ammonium and ammonium salts, salts, among among numerous numerous other other acids acids and bases and bases well known well known in the pharmaceutical in the pharmaceutical

art. art. Sodium and Sodium and potassium potassium salts salts are particularly are particularly preferred preferred as neutralization as neutralization salts ofsalts the of the

phosphates according phosphates accordingtotothe the present present disclosure. disclosure. term term

[0446] TheThe

[0446] "pharmaceutically "pharmaceutically acceptable acceptable derivative" derivative" is used used throughout is throughout the the specification specification to to describe describe any any pharmaceutically acceptable prodrug pharmaceutically acceptable prodrugform form (suchas asananester, (such ester,amide amide other prodrug other prodrug group), group), which, which, upon administration upon administration to a provides to a patient, patient, directly providesordirectly or the indirectly indirectly the present compound present compound or or anan activemetabolite active metaboliteofofthe thepresent present compound. compound.

[0447] General

[0447] Synthetic Approach GeneralSynthetic Approach

[0448] The synthetic

[0448] The synthetic realization and and realization optimization optimization of the the bifunctional of bifunctional molecules molecules as described as described

herein may herein maybebeapproached approached in ainstep-wise a step-wise or modular or modular fashion. fashion. For example, For example, identification identification of of compounds thatbind compounds that bind to tothe thetarget targetmolecules moleculescancan involve involve high high or or medium medium throughput throughput screening screening

campaigns if no campaigns if no suitable suitable ligands ligands are immediately are immediately available. available. It is notItunusual is not for unusual initialfor initialtoligands to ligands

require iterative require iterative design design and and optimization cycles to optimization cycles to improve improvesuboptimal suboptimal aspects aspects as as identifiedbyby identified

data from suitable data from suitable in in vitro vitro and andpharmacological pharmacologicaland/or and/orADMET ADMET assays.assays. Part Part of the of the

optimization/SAR campaign optimization/SAR campaign wouldwould be to be to positions probe probe positions of thethat of the ligand ligand are that are of tolerant tolerant of substitution substitution and and that that might might be be suitable suitable places places on on which to attach which to attach the the linker linker chemistry chemistry previously previously

referred to referred to herein. herein. Where crystallographic ororNMR Where crystallographic NMR structural structural datadata are are available, available, these these can can be be used tofocus used to focussuch such a synthetic a synthetic effort. effort.

209

2023248067 10 2023

[0449] In a Invery

[0449] a very analogous analogous waycanone way one can identify identify and optimize and optimize ligandsligands for an for an E3 Ligase, E3 Ligase, i.e. i.e. ULMs/CLMs. ULMs/CLMs. Oct [0450] With

[0450] PTMsPTMs With and ULMs and ULMs (e.g. CLMs) in handin one (e.g. CLMs) hand one skilled skilled in the art can the can in art use use known known

synthetic synthetic methods for their methods for their combination withororwithout combination with withouta alinker linkermoiety. moiety.Linker Linkermoieties moietiescan canbebe synthesized with synthesized with a range a range of compositions, of compositions, lengthslengths and flexibility and flexibility and functionalized and functionalized such that the such that the

PTMand PTM and ULM ULM groups groups canattached can be be attached sequentially sequentially to distal to distal endsends of the of the linker. linker. Thus Thus a libraryofof a library

bifunctional bifunctional molecules canbeberealized molecules can realized and andprofiled profiledinin in in vitro vitro and in vivo and in vivo pharmacological pharmacologicaland and ADMET/PK ADMET/PK studies. studies. As with As with the and the PTM PTMULM and ULM the groups, groups, finalthe final bifunctional bifunctional molecules molecules can be can be subject subject to to iterative iterativedesign design and and optimization optimization cycles cycles in in order order to to identify identifymolecules molecules with desirable desirable properties. properties.

Abbreviations:

[0451] Abbreviations:

[0451] ACN: acetonitrile ACN: acetonitrile

AcOH, aceticacid AcOH, acetic acid ADDP: 1,1'-(azodicarbonyl)dipiperidine ADDP: 1,1'-(azodicarbonyl)dipiperidine

aq., aq., aqueous aqueous

BAST: N,N-bis(2-methoxyethyl)aminosulfur BAST: N,N-bis(2-methoxyethyl)aminosulfur trifluoride trifluoride

BINAP, 2,2'-bis(diphenylphosphino)-1,1'-binaphthalene BINAP, ,2,2'-bis(diphenylphosphino)-1,1'-binaphthalene

Boc, tert-butoxycarbonyl Boc, tert-butoxycarbonyl

Boc20, di-tert-butyl decarbonate BocO, di-tert-butyl decarbonate BOP, (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium BOP, (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate hexafluorophosphate

BPO: benzoylperoxide BPO: benzoyl peroxide Cbz: Cbz: Carbonylbezyloxy Carbonylbezyloxy

CDCl , deuteriochloroform CDCl, 3 deuteriochloroform

CD30D, deuteriomethanol CD3OD, deuteriomethanol

CH 3CN, CHCN, acetonitrile acetonitrile

CH 30H,methanol CHOH, methanol CsF, cesiumfluoride CsF, cesium fluoride Cs2CO3, cesium CsCO, cesium carbonate carbonate

Cu(OAc)2, copper Cu(OAc), copper (II)acetate (II) acetate Cy2NMe, dicyclohexylmethylamine CyNMe, dicyclohexylmethylamine

DAST:diethylaminosulfur DAST: diethylaminosulfur trifluoride trifluoride

210

DBE: 1,2-dibromoethane DBE: 1,2-dibromoethane

DCM: dichloromethane DCM: dichloromethane

DEAD: diethylazodicarboxylate DEAD: diethyl azodicarboxylate DIAD: diisopropylazodicarboxylate DIAD: diisopropyl azodicarboxylate DIBAL: disiobutylaluminium DIBAL: disiobutylaluminium hydride hydride

DIEA or DIPEA: DIEA or DIPEA:disopropylethylamine diisopropylethylamine DMA: N,N-dimethylacetamide 2023248067

DMA: N,N-dimethylacetamide

DMAP, N,N-dimethylaminopyridine DMAP, N,N-dimethylaminopyridine

DMF: N,N-dimethylformamide DMF: N,N-dimethylformamide

DMP: Dess-Martin periodinane DMP: Dess-Martin periodinane DMSO, dimethylsulfoxide DMSO, dimethylsulfoxide

DMSO-d hexadeuterodimethyl DMSO-d, 6, hexadeuterodimethylsulfoxide sulfoxide EA: ethyl acetate EA: ethyl acetate EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodimide

Et 2NH,diethylamine EtNH, diethylamine EtOAc EtOAc ororEA, EA,ethyl ethylacetate acetate HCl, hydrochloric acid HCI, hydrochloric acid H 20, water HO, water

HBTU: N,N,N'N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate HBTU: N,N,NN'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate HMDS: bis9trimethylsilyl)amine HMDS: bis9trimethylsilyl)amine

HMPA: hexamethylphosphoramide HMPA: hexamethylphosphoramide

HPLC, highperformance HPLC, high performance liquid liquid chromatography chromatography

IBX, 2-iodoxybenzoic IBX, 2-iodoxybenzoicacid acid KOAc, potassium KOAc, potassium acetate acetate

LCMS, liquidchromatography LCMS, liquid chromatography / mass / mass spectrometry spectrometry

LDA: lithiumdiisopropylamide LDA: lithium diisopropylamide LiOH,lithium LiOH, lithiumhydroxide hydroxide MCPBA: meta-chloroperoxybenzoicacid MCPBA: meta-chloroperoxybenzoic acid MeOH, methanol MeOH, methanol

MsCl: methanesulfonylchloride MsCl: methanesulfonyl chloride M.W: microwave M.W: microwave

211

nitrogen N 2,nitrogen N,

NaH, sodium NaH, sodium hydride hydride NaBH 3 CN, NaBHCN, sodium sodium cyanoborohydride cyanoborohydride

NaBH(OAc)3, NaBH(OAc), sodium sodium triacetoxyborohydride triacetoxyborohydride

NaCl, sodium NaCl, sodiumchloride chloride NaHCO3, NaHCO, sodium sodium bicarbonate bicarbonate

Nal, sodium sodiumiodide iodide 2023248067

Nal,

Na2SOsodium NaSO, 4 , sodium sulfate sulfate

NBS: N-bromosuccinimide NBS: N-bromosuccinimide n-BuLi, n-butyllithium n-BuLi, n-butyllithium NH 3,ammonia NH, ammonia NH 4Cl,ammonium NHCl, ammonium chloride chloride

NH 2 OH-HCl, NHOH·HCI, hydroxylamine hydroxylamine hydrochloride hydrochloride

NMP,N-methylpyrrolidone NMP, N-methylpyrrolidone NMR,nuclear NMR, nuclear magnetic magnetic resonance resonance

02,oxygen O, oxygen

PCC: pyridiniumchlorochromate PCC: pyridinium chlorochromate Pd-118orPd(dtpf)Cl : 1,1'-bis(di-tert-butylphosphino)ferrocenedichloropalladium Pd-118 or Pd(dtpf)Cl: 21,1-bis(di-tert-butylphosphino)ferrocene dichloropalladium

Pd(aMPhos)C12, bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) Pd(aMPhos)Cl, bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(Il)

Pd 2(dba) 3Tris(dibenzylideneacetone)dipalladium Pd(dba): : Tris(dibenzylideneacetone)dipalladium Pd(dppf)C121,1'-bis(diphenylphosphino)ferrocene Pd(dppf)Cl: : 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium dichloropalladium

Pd(dba) Pd(dba):2 :bis(dibenzylideneacetone)palladium bis(dibenzylideneacetone)palladium Pd(OH) Pd(OH),2 ,palladium palladium hydroxide hydroxide Pd(PPh Pd(PPh),3)tetrakis(triphenylphosphine)palladium(0) 4 , tetrakis(triphenylphosphine)palladium(O)

PE, petroleumether PE, petroleum ether Ph 3 P,triphenylphosphine PhP, triphenylphosphine PPTS: pyridiump-tolunesulfonate PPTS: pyridium p-tolunesulfonate PTSA:p-toluenesulfonic PTSA: p-toluenesulfonicacid acid Py, pyridine Py, pyridine

PyBOP, (benzotriazol-1-yloxy)tripyrrolidinophosphonium PyBOP, (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate hexafluorophosphate

rt, rt,room room temperature temperature

212

RuPhos-Pd-G3: RuPhos-Pd-G3: XPhos-Pd-G3: XPhos-Pd-G3: [(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)-2

[(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)-2-

(2'-amino-1,1'-biphenyl)] palladium(II) methanesulfonate (2'-amino-1,1'-biphenyl)] methanesulfonate

RuPhos-Pd-G2: RuPhos-Pd-G2: Chloro[(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)-2-(2' Chloro[(2-dicyclohexylphosphino-2',6'-disopropoxy-1,1'-biphenyl)-2-(2'-

amino-i,1'-biphenyl)] palladium(II) amino-1,1'-biphenyl)] palladium(II)

SFC:supercritical SFC: supercritical fluid fluid chromatography chromatography

TBAF,tetra-n-butylammonium TBAF, tetra-n-butylammonium fluoride fluoride

TBDPSCl, tert-butyldiphenylsilylchloride chloride 2023248067

TBDPSCI, tert-butyldiphenylsilyl

TBS,tert-butyldimethylsilyl TBS, tert-butyldimethylsilyl tBuOK,potassium tBuOK, potassium tert-butoxide tert-butoxide

[tBu 3PH]BF4,

[tBuPH]BF, tri-tert-butylphosphonium tri-tert-butyl phosphonium tetrafluoroborate tetrafluoroborate

t-BuXPhos-Pd-G3: t-BuXPhos-Pd-G3: [(2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino

[(2-di-tert-butylphosphino-2',4',6'-trisopropyl-1,1'-biphenyl)-2-(2'-amino-

1,1'-biphenyl)] 1,1'-biphenyl)] palladium(II) palladium(II) methanesulfonate methanesulfonate

TEA: trimethylamine TEA: trimethylamine TFA:trifluoroacetic TFA: trifluoroacetic acid acid

TLC:thin TLC: thin layer layer chromatography chromatography TMP:2,2,6,6-tetramethylpiperidine TMP: 2,2,6,6-tetramethylpiperidine TEMPO: TEMPO: 2,2,6,6-tetramethylpiperidine-N-oxide 2,2,6,6-tetramethylpiperidine-N-oxide

TMSOTf, trimethylsilyltrifluoromethanesulfonate TMSOTf, trimethylsilyl trifluoromethanesulfonate TosCloror TsCl: TosCl TsCl: p-toluenesulfonyl p-toluenesulfonyl chloride chloride TsCl, p-toluenesufonyl TsCl, p-toluenesufonyl chloride chloride TsOH: p-toluenesulfonicacid TsOH: p-toluenesulfonic acid XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

XPhos:2-dicyclohexylphosphino-2'4'6'-triisopropylbiphenyl XPhos: 2-dicyclohexylphosphino-2'4'6'-triisopropylbiphenyl XPhos-Pd-G3: [(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1' XPhos-Pd-G3: [(2-dicyclohexylphosphino-2',4',6'-trisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-

biphenyl)] palladium(II) methanesulfonate biphenyl)] palladium(II) methanesulfonate

12354-85-7: bis(pentamethylcyclopentadienylrhodium 12354-85-7: bis(pentamethylcyclopentadienylrhodium dichloride) dichloride)

[0452] A. A.

[0452] Exemplary Exemplary Synthetic Synthetic Schemes Schemes for Exemplary for Exemplary Estrogen Estrogen Receptor Receptor Binding Binding

Moiety Moiety Based Based Compounds Compounds Synthetic

[0453] Synthetic

[0453] scheme scheme A-2 through A-1,through A-1, A-2 A-5,and A-5, A-6, A-6, A-7and A-7 described described the used the routes inused routes in the preparation the preparation of of CRBN ligands,asaswell CRBN ligands, wellasasCRBN CRBN ligands ligands with with partial partial linkermoieties linker moieties connected. connected.

213

Oct 2023 104541 General

[0454] General synthetic synthetic scheme A-1 toA-prepare scheme I to prepare intermediate. intermediate.

L"OH L L "'OH OH LK"OH OH OH L"OH LK"-0 O OH F XIi F NaNO2HBr, CuBr , HBr,CuBr 6Dess-Martin X YY IZ N Pd/C, HH2 N NaNO, Dess-Martin

N Pd/C, N _______ NN N N 0 2023248067 10

K20, M X_( X Y THIF THF X Y ' H 2 0,O0-100 HO, 0-100 °CC x X Y X Y KCO, DMF NO 2 NO 1:. NO 2 NH2 NH Br Br Br Br NO o OTf o 0i~ 0OTf O O o o o' B-B Bn o pyridinium tribromide TsOH TsOH N Pd(dPPf)C1 2, KOAc Pd(dppf)Cl, KOAc N Pd(dPPf)C1 2, KKCO 2C0 3 N pyiiimrirmd N Pd(dppf)Cl, N N N trimethoxymethane X trimethoxymethane dioxane, 100 °C dioxane100 OC x N,:WY x X Y yx y X Y X X Y dioxane/H 100 0°C 0, 100 dioxane/HO, 2 OC I TEA, DCM, 0-25 °C, 2h TEA, DCM, 0-25 C, 2h I Br Br Or B N N Br Br Bn o Bn o

"1 O 01 o N 0 0 o o L <-O o L>"0OL- o o

N N N Pd(dPPf)C1 2 , KKCO Pd(dppf)Cl, 2C0 3 N P/HNN chiral SFC separation chiral SFC separation N N x N xdC N y Pd/C, H H2 xX Y X + X Y X Y TFEO THF/EtOH Y dioxane/H20, dioxane/H2O, 100OC 100 °C

phenylboronic acid phenylboronic acid

Bn, HO HOIDHO Ho Bn HO o HO X and Y X and Y = =H, F; L(CH ) where n 0,1,2 H, F; L = (CH) where 2 n = 0, 1, 2

General 104551 General

[0455] synthetic synthetic scheme scheme A-2 toA-2 to prepare prepare intermediate. intermediate.

214

Oct 2023

'0 OH Br"-Br rN 1 H NH 0 0 N 0 0 OH Br Br N 0 ~ 0 o Boc" N,_0 Boc NaOH NaOH N. OH OH o `.Zz OH Pd(OAc) (0.1 eq), KHPO (3 eq) FPd(OAc)2 (0.1 eq), K 2HPO4 (3 eq) 0 DIPEA, DIEA, DMSO DMSO NN H 2 0, MeOHITHF- rN'( OH F HO, MeOH/THF N dioxane, 140 °C, 10 h F 120 °C, 12 h N N diloxane, 140 *C,l10h F 120 0, 12 h Boc Boc N Boc' Boc

2023248067 10

0 o NH O o DIEA O o0 MnO 2 MnO o 0 H2N HI 0. O HATU, DIEA HATU, H. O HCI -OHN.N OH NH OH H N o 0 25 C,h DCM,25°C, DCM, 1h I 0 NN DMFC N N DMF 0 N N ~ O NaOAc, NaBHC NaOAc, NaBHCN N N Boc N H AcOH,MeOH AcOH, MeOH NJc N 0I~ NN0J IZ N Boc H Boc o N O Boc H

HCI/Dioxane HCI/Dioxane

\0 O 1 0 HCI 'N N \- ,,N_ N NH NH HN H 0 o

104561

[0456] General synthetic General synthetic scheme A-3totoprepare schemeA-3 intermediate. prepareintermediate. 0 o 0 0 0 o 0 o Brr,,-0 o

X OH H2SO H S0 - FFFFF -Ci 0~ NaOH NaOH e OH OH 2 4 N. N.F F 0F F 0 O F j F OH OH MeOH, MeOH, 85C,20 85°C, 20 h h F F OH 1(3003,DMF KCO, DMF THF/MeOH/H 20 THF/MeOH/HO OH 80-C, 12h 80°C, 12h F1 F F 'NlFF F F F

FF FF F F N_H) F N0< OH F"' O ' i F ~ NH F o OH F N O Boc N. 0CH 3CH 2 , K2C03 CHCHI, KCO 0__0______N__0 o AgOAc, cat: 12354-85-7 O O DIPEA, DMSO, 120°C, 12 h 1-methylnaphthalene F" N.rFN DMF, 20°C, 1h N 1-methylnaphthalene F F 0 100°C, 12h 100 C,12 h FBoe NNN Boc

FF O O F F 0 o F F H2N 'C H2N NH F 0 0F 0 00 NH F o F o o F o 0S04(2% eq), NaIO (3 eq) N. 0 DIPEA N NH OsO(2% eq), NalO (3 4eq) o0- DIPEA N.0 1N N 0 3CN NN , N DMF - IA: THF, H2O, 20°CTH, 2 ,0CNaOAc, NaOAc, NaBH NaBHCN -ll DMF N N DCM,MeOH DCM, MeOH ~ N N NH N N N BoO NN N Boc Boce o Boc' Boc Boc

F F

HCI/dioxane HCI/dioxane F'"0 F 0 0 o 0-0 NH dioxane N HCI N HNJ HN

104571

[0457] synthetic scheme General synthetic General schemeA-4 prepareintermediate. A-4totoprepare intermediate.

215

0~~ 0NHBc NH o CU2 O,NaOH o CHI, K 2CO3 o Bo N 0 o NIS Boc NIS CuO, NaOH CH|, KCO O Br Br ~ trifluoromethanesulfonic trifluoromethanesulfonicacid acid Br BrH 2 0,DMF, HO, 80°C DMF,80 OC Br Braeoe acetone Br r IOH OH 0, o 0 o 0 0 0 o o o NaOH NaOH - ~ OH OH Mn2HCH2- OH CIH HN NH =0OCOCH0 MnO COOH N THF, H NTHF, H2O0 N OH OH NNO)II o H o 2 INO NaOAc, NaBHCN, N DCM DCM N,00 NaOAc, NaBH 3CN, IN_ NH N o N -P br-t o H N NH Boc' Boc Boc Boc O Boc' N Boc o MeOH Boc HMeON Bc[ , N Boc o o 2023248067

o00 0 o0 o 0 NH NH HATU _____ IN _C) N N 0 o HCI/dioxane r NC) IC/ixn. N-L N 0 o ri~ N N DIPEA, DMF DIPEA, DMF dioxane Boc' N Boc N,_) o 0, dioxane HN HN") o 0" HCI HCI

o o $l NH o 00 o 0 0 NH Br N NaOH o Boc 0oc IN0__ ,_ _ o NaOll- OH Br 0rr NO OH Br Br N MeOH, N KCO, DMF Br6K20, M XPhos-Pd-G3, CsCO, XPhos-Pd-G3, Cs2 CO 3, MOH,THF, THF,HOH 2 0OP OH o 0-' - ` dioxane, 90*C, dioxane,90 16 hh °C, 16 BocN BocN ) o 20°C, 2h h Boc IN, ,_-,,N 20-C, 20C OH o Boc o o O o MnO 2 NH OHO0 COOH NH OH CIH HN o HATU, DIEA MnO O Mn02OH IH- H H 0 N HATU, DIEAC; N- N o DCM, 20 °C, 2 h N N N N H DMF, . NH 1 hh 20C,1 DMF,20°C, r N NaOAc, NaBHCN 0,_ 0Boc-O Boc BN N oI 0_,,, NaOAc,NaBH 3CN Boc IN N oO.. 0 ,-, O Boc N o MeOH, 20°C, 2h O 0 0 o NH NH HCI/dioxane HCl/dioxane IN 0 N o N i::: t. HC N DCM HN HCI HN J o 0,_-,, HCI

104581 General

[0458] General synthetic synthetic scheme A-5 toA-5 scheme to prepare prepare intermediate. intermediate.

216

Oct 2023 0 0 0 0 o o o o b Br- Br'-'O..O-. N BFJK NaOH N OH OH BFK NaOH o KCO,3 , Pd(dPPf)C1 2, KCO N o H2 0,20°C, TH F, HO, 200C,l1h N- OH Br Br K2CC3 KIKI KCsCO, Br C6 Br Pd(dppf)Cl, THF, 1h -

OH DMF, 100°C, 10000C,lh O,,-0 .O Dioxane, Dioxane,H 2 0,700, 70°C, 3h 3h OH DMF, 1h o HO, o0----0,o-oo o

0 o H2N H2N 0 OH 2023248067 10 OH MnO 2 o H HNONNH0 0 NH H o DM,2,l 0 HN H2N o N, N. NH2 HATU,DIEA HATU,DIEA NN o NH2 MnO OH o NH DCM, 20°C, 1h OH,20C NaBH3CN,AcOH NaBHCN, AcOH -0,N-N---- o DMF,20°C, DMF, 2000, 1h lh N° o MeOH,20°C, 201C, 12.5h 12.5h MeOH, 0 o 0 0----,- o O OLOO

0 NH2 0 NH2 o NH o NH Cbz, N--\ Cbz-

I o o N NaBO 3.4H NaBO 2O, BH 4HO, 3 -Me 2S BH-MeS N- N° TosCi, DMAP, TosCl, DMAP, TEA TEA I N" ,N N° NH THF, 0-200C,3h3h THF, 0-20°C, Ho HO ~ ~ DCM, 50TODIPEA, 25°C, 12h TsO ACN, DIPEA, ACN, 60-C, 60°C, 24h24h

O O O o

o o 0 O 00 O o o o <"N N° o ~ Cbz-N N-'\- 0 N.N N I'- N 0 Cbz- N N N N',. N,,, NHHN NH HN N \ N,,, NH Cbz-N\ Cbz-N o 2 H2N o >0 o P/,15 Psi Pd/C, H, 15 Psi 0~ o PdCH2 0 fo HN benzensulfonic acid benzensulfonic acid 0 70

o DMF, 20°C, 2h ACN, 80°C, 12h 0ACN, 800, 12h o 0 DMF, 200C, 2h o 0

o o o

104591 General

[0459] General synthetic synthetic scheme scheme A-6 A-6 to to prepare prepare intermediate. intermediate.

0 0 0 0 o o o o 0 o "N NH o NH 0 Br NHH o 0 S Boc' Boc N / TiiPO4 Ti(i-PrO) Br o N DIPEA, DMSO DMSO EtO, EtMgBr F F C NDIPEA, rNN Et 2O, EtMgBr N NaH,DMF, NaH, DMF,303000C,12h °C, 12h N N N Boc/C Bac/N Boc Boc

0 O 0 O / NH 3 H2 0/MeOH NH'HO/MeOH o 0o O JCN N 0 (1) 50 °C, 15 h (1) 50 C, 15h CI- NtNH N N 'N N 0 HN,_ HN 0 O OH BoN N Bc'T / (2) (2 -,\\0 S Boc 0 003.0 o eq 3.0 eq 0 o MeCN, MeCN, 909000C °C, 2h2h

104601 General

[0460] General synthetic synthetic scheme scheme A-7 A-7 to to prepare prepare intermediate. intermediate.

217

Oct 2023

N-Boc Boc N HH0 2 CDI CDI HN HN\ N-Boc BocN NH H2O N N LC,2h 20 °C, 12 hr N~ NNNH RuPhos-Pd-G2, tBuN MeCN, 50 °C, 12h N N FF F 20 ',12 hr N N IZ N H0 NH NH2 MeCN, 50 °C, 12h NHR hP-Gt RuPhos-Pd-G2, u t-BuONa N N N H o t-Amyl-OH, t-Amyl-OH, 90 90°C, °C, 16h 16h N N o N NH 2023248067 10 Br NH 00 N HN NYNH NH Br--Z o O O HN HN) O Br o N OHCI/Dioxane NH HCI/Dioxane o o Boc NH N NBoc N N N o NN NH tBuOKMeCN 'BuOK, MeCN, 12 2hr hr N 7C Nz N HCI <N N0N o DM HI N 1000°C 100 O NC- N DCM 0 O 0 o

[0461] Synthetic

[0461] Synthetic schemes schemes A-8, A-10, A-8, A-9, A-9, A-10, A-11, A-11, A-12, A-14, A-12, A-13, A-13, A-15, A-14,A-16, A-15,and A-16, A-17,and A-17, described the routes described the routes used used in in the thepreparation preparation of ofrepresentative representativechimeric chimericcompounds claimedinin compounds claimed

this this application. application.

General

[0462] General

[0462] synthetic synthetic scheme A-8 toA-8 scheme to prepare prepare claimed claimed compounds. compounds.

HO Ho A NN O N N HN N-Q\ o N H NaCNH HOAc, MeOH NaCNH,3 ,HOAc, HN N MeOH O N N __ _ _ _ _ _ O + N + HCI N IZ Y O N O HIH H HO R R O O Ho HN Y Y R HN R O n =0, 2, 3, 4 n=0,2,3,4 O- H H o R=HF; R = H, F; Y=CH,N Y=CH,N

(CH2)n HO O N N Ho O N O HN N N NaCNH 3 ,HOAc, NaCNH, HOAc, MeOH MeOH O o o N + N- N o0N _

HCI o O o IZ N O Y H H RO R o HOH Ho HNQ HN Y R H O O n=0,2,3,4 n=0,2,3,4 R=H,F; R = H, F; YY=CH,N = CH, N

General

[0463] General

[0463] synthetic synthetic scheme A-9 toA-9 scheme to prepare prepare claimed claimed compounds. compounds.

218

Oct 2023

0HH2N 2N O :- H 2N H2N 0H o 2N H2N o 0 "BFBF3 K 0 0r NaB 3 .41H,BH3 -Me 2 S 0 TosC,ODMAP, TEA K o NaBO 4HO, BH-MeS TosCI, DMAP, TEA o N" 0 No 0 / ,dPP)'2, KCO Pd(dppf)Cl, 2- N" 0,,, o THF.O-15 THF, C,3h 0-15°C, 3h N0 CM,O0-15 DCM, C,.2h 0-15°C, 2h

o dioxane, HO, 70 °C, 3h Br Br 0 dioxane, H 2 , 70 C, 3h o 0 HO Ho 0 o o H2N (NHNH O 0 0 O NH 2 0oH 2N o o 2023248067 10

NH NH N 0 Cbz1 N N'z'_ Benzenesulfonic Benzenesulfonic acid acid o N° NH No N° o KI, DIPEA, KI, DIPEA,CH 3 CN CHCN 0 CH CN 3 N o CHCN Cbz-N TosO TosO o0 N O 0~' X-\ Cbz\-j o o ~~CbzNi_ Cbz-NN

HO"-- H H2, Pd/C H2, Pd/C HO THF/Et0H THF/EtOH

N HO,,; Ho N O0 o \NN0\ o N,, NH 0IM N,, 7 HN HNNNN o N\ NH N,, NH N0 o MeOH, DCE, AcOH, NaBH3CN 0 MeOH, DCE, AcOH, NaBHCN o

104641

[0464] General synthetic General synthetic scheme schemeA-10 A- 1 totoprepare prepareclaimed claimed compounds. compounds.

0 0 o 0 (1) TMSCHN (1) 2 , ACN TMSCHN, ACN o CI Cl Co ~(2) ~Br0 Br o (2) HBr/AcOH, HBr/AcOH, ACNM ACN o o o 0 o OH o 0 Bn 0o N o0 o" IFEI Bn1 BnN Br Br 0" o Bn -k 0 x0 DAT- o 0 o FF DAST Bn o o N , acetone-N K 2C0 3acetone KCO, DCE DCE o o

Bn Bn O CIH.HN N LiAIH4

\/~ F o o FCHH' NBNN oFF NH 0 Bn- 0 IF Bn o o F F o Bn o THF

F NN (1) chiral SFC o

N N N 0 o NaOAc, NaBH CN, AcOH, MeOH 3 NaOAc, NaBHCN, AcOH, MeOH I (2) PCC, (2) PCC, DCM/0F DCM 0 F o F N NH NI Pd/C, HH2 Pd/C,

HO THF THF o 0 OH HO 0\ FF F - o 00o N N NH N O

104651 General

[0465] General syntheticscheme synthetic schemeA-11 A-i 1totoprepare prepare claimed claimed compounds. compounds.

219

Oct 2023

00 0 o o 0 ONa ONa o H(II>0 OH NaBH HCI 00 011111 Na0H,MeOH, NaOH, MeOH, o reflux reflux 0 P1)IA 1 0 o HCI HO TsOH, toluene TsOH, toluene o co\0 0 / Y NaBH4 MeOH MeOH I

. NaO NaO 0 o 0 -00o 2023248067 10

TsCI - SC TsO- T v0 HO---T- TsO Ho DOM, TEA \---- b D DCM, TEA -

Bn- Bn 0O Bn . 0*, Br<O 0~ Bn OH OH Bn IN \S -- o (1M)0 HCI (1M) HCI o TsO o THF F HI CS2CO 3,DMF CsCO, DMIF F F I F FF 00 0 f-\- 0N~N NH N N NH N N N Ho HN o O o HCI HCI 0 0 o Pd/C, HH2 Pd/C, N' a 3 O THE NaBHCN, NaOAc, MeOH/DCM THF N NaBH 3CN, NaOAc, MeOH/DCM IF F FF HO HO

220

[0466] General synthetic General A-12to toprepare schemeA-12 synthetic scheme claimed prepareclaimed compounds. compounds.

F F x IF F F F t-Bu OH o F O o X OH~ I O~ I F, F F, F 0x S HN F 0FF F F o F F F 0 S F F O F o O F F F X X KCO, THF, MeCN t-BuONa, Pd(OAc) t-BuONa, 0dO~) K2C0 3,THF, MeCN t-Bu t-Bu 0dO~) t-u~I XPhos toluene, 90°C \/Bu N Noun,9* o XPhos I o t-Bu o 2023248067 00

HO HO X o o NH ax N HO N HO =0 N N N 0I HCI HSO(2M) X HN N o THF(2M) H 2S0 4 \/ N NaOAc, 0xN_ NaBHCN, IO MeOH, DCM aHC, eH C THIF NHjN)c X=H, F X= H,IF N N o o o N NH NH NH N DMH MeOH, DCM HN HNN I N 0 NaOAc, NaBH NaOAc, 3 CN, MeOH, NaBHCN, DCM HCI NH 0 o 0 o o

HO xI X N HO N N N N

0 o N O o I XH,F NH X=H,F 0 O

General 104671 General

[0467] syntheticscheme synthetic to prepare claimed A- 13to prepare schemeA-13 compounds. claimed compounds.

221

OH CbzCI OH DMP /0 CH(OMe) . 0- H,, Pd/C ON-Y0 CH(OMe)3 Oct 2023 OH CbzCl OH DMP O H, Pd/C o HN N HN Na2 003DCM, NaCO, ,0DCM,H2O H2 0 DCM,rk DCM, rt TsOH, MeOH TsOH, MeOH ~.MeOH, MeOH, THEF THF HN, HN 0 NN IFIFIFCb Cbz NN Cbz'N F F F Cbz Cbz F F F F FF Bn OH Ho OH HO N OS F Bn O O 2023248067 10

O o O \ -HN HN \/Pd/C, Pd/C, HH(50 2 (50Psi) psi)

__ t-BuOK, Pd(OAc) t-BuOK, Pd(OAc) 2 / \/ N MeOH,THF MeOH, THF N N chiral SEC chiral SFC separation Ho H + XPhos XPhos 0eaato n tOoluene,90'C / spartioHO toluene, 90°C N Bn O SOH OH o 0, Ho OH N NHO OH o NH N HSO H 2 S04(2M) (2V) N =0 - N H THF, 70 °C N \/ND - HN., HCIC HN N TH, 70 OC o NaOAc, HOAc, MeOH, NaOAc, HOAc, MeOH, DCM DCM j N N

O Oo NaOAc, HOAc, o ON NaOAc,DCM MeOH, HOAc, HN 'N N C N y~ NH N Nh N HCI o N HO HI0Oo Ho o o N\: NIN/N NH N N N N N O 4H NH 0 0 NH o 0 O

104681 General

[0468] General syntheticscheme synthetic claimed compounds. prepare claimed A-14toto prepare schemeA-14 compounds.

222

Oct 2023 t-BuO t-BuO -~OH OH

OBn OBn OH OH N2

0 Pd/C, Pd/C,HH2 O _ TosCi, KOH TosCI, KOH 0~ OTss -

H /- o DMFTH- CS2 CO 3 ,DMF THF, EtOH THE, EtOH /- THF CsCO, 2023248067 10

t-BuON t-BuO

O OH 00'OH OH OH O O AIUiH AILiH4 SEC separation SFC separation

THE THF

tB t-BuO~~ t-BuO B 0 H ZI t-BuO t-BuO t-Bu0 t-BuO O O NHN N OH000 Nc o N OH J~ O N rOH I NNHNH N-,) N N N O O HN o Dess-Martin N HCI 0I N Dess-Martin O - DCM NaQAc, NaCNBH 3 DCM NaOAc, NaCNBH N N' DCM/MeOH, DCM/ MeOH,rt,rt, 1212h h

" t N t-Bu0N t-BuO BuO BuO t-BuO( t-BuO

O 0< N ~N 0 NH N N HCI/dioxane HCI/dioxane HO\ NH Ho 0 DCM DCM

104691 General

[0469] General syntheticscheme synthetic schemeA-15 A- 15totoprepare prepare claimed claimed compounds. compounds.

223

Oct 2023

Bn OH OH A o~l TosCl, TEA DCM DCM Bn AE OTos OTos

o Bn Bn OL~iiY iirOH OH OH OH OTos

Bn-0 o o O 02 dC 5piP Ph Ph 0 O H2, Pd/C, 15 psi N H 2,Pd/C,15psi Swernoxidation N Swern oxidation ~ Ph P~~j 2023248067 10

I Cs 2CO.DMVF, CsCO, 80C, DMF, 80°C, 16h 16h MeOHITIHF, MeOH/THF, r.t., 16 h Ph DCM, r.t., 6 h N r.t. ,16 h - ~DCM, r.t.,6 h t-BuO t-BuO N t-BuO t-BuO Nt-BuO N t-BuO t-BuO t-BuO

OH 0 OH O

OH OH 0"- OH OH 0'L o o o o OH NDIBAL-H DIBAL-H Nm-CPBA N LAIH- LiAIH4 4 NDMVIP m-CPBA DMP - DCM,-78°C DCM, -78C ~ ~DCM, r.t,1616h DCM, r.t., h ~~ THE DCM DCM THF

t-BuO t-BuO Nt-BuO t-BuO Nt-BuO t-BuO Nt-BuO t-BuO N

-0 O OH - 1- IN 00l

OH N N - OH OH o N NH N IZ N N OH N N o =0 OH o o0 N N N 0N HN o HN)0HCI/dioxane HCI/dioxane N 0 H IZ

TEA, NaBH(OAc) CM, TEA, D-DCM, NaBH(OAc) 3H

t-BuO' t-BuO HON HO

)0

104701 General

[0470] General syntheticscheme synthetic schemeA-16 A- 16toto prepare prepare claimed claimed compounds. compounds.

OH

CI NaBH4, CaCl N SOCI N HO OH CI CI MeOH, 0-20 °C, 16 h N THF,80°C,4h H OH o N CsCO, ACN, 90°C, 3h

FIN N ON O 0aH4 NB1 01 H HO Z N /cl Ncpl/ 0EIOFt11 0 M.H NN2'C 16 h THF,20*C,3h N CSC3 NN 0C,3

H H HN N N o 0- 0-H TFA N N° N N 10471 TEA, KI, DMF, r.t, 16h Geea syteiHceeA1 N N opeaecamdcmons o THF, 20°C, 3h N

O

[0471] General synthetic scheme A-17 to prepare claimed compounds.

HN N 0 0 02N4

Oct 2023 0 BnO BnO OH OH 'I'OA -I OH OH o

o o 0 O LLAIH 4, THF LIAIH4, THF Dess-Martin Dess-Martin

DCM, DCM, 0 °COC to r.t.,1h1 0 to r.t., h CsCO, DMF, 80°C, 16h 16h 80C, 0°C, 11hh 0°C, Tos-O Cs 2CO3, DMF, Tos o 2023248067 10

BnO / BnO BnO BnO BnO BnO 0 0 o HO Ho o Br Br Br

O O o Br Br Br O~~O m-CPBA m-CPBA LiBr, LiBr,AcOH/THF AcOH/THF Dess-Martin Dess-Martin DCss-rtin DCM, 0 - r.t., 1h \ DCM, 0 C to r.t., 16 h DCM, 0 °C to r.t., 16 h rt, 16 h r.t., 16h

n-BuLi, -60°C,°C,2h2 h n-BuLi, -60

BnO BnO BnO BnO BnO BnO BnO BnO

0 o 0 O 0 1 N N 0 NH o N _NH O 000 NI H=0 N N o 0 0_1 N NN0\' NH 0 HN.)N 0 N /_NN

/ o o N HN H 30 N NJ Pd/C, H ,50 PSI Pd/C, H, 50 2 psi

ACN/DMSO, ACN/DMSO, DIEA, DIEA, 70 'C,2 2h 70 °C, h 110- HO 0'N10 o N o DMF/THF, 6h 0OMF/THF, 6h - /H H

BnO BnO

[0472] Exemplary

[0472] Exemplary Synthesisof ofExemplary Synthesis Exemplary Compound Compound 2: 3-{5-[4-(5-{4-I(1R,2S)-6 2: 3-{5-[4-(5-{4-[(1R,2S)-6- hydroxy-2-phenyl-1,2,3,4-tetrahydronaphthalen-1-yl]phenoxy}pentyl)piperazin-1-yl]-7 hydroxy-2-phenyl-1,2,3,4-tetrahydronaphthalen-1-ylphenoxy}pentyl)piperazin-1-yl]-7-

methoxy-1-oxo-2,3-dihydro-1H-isoindol-2-yl}piperidine-2,6-dione methoxy-1-oxo-2,3-dihydro-1H-isoindol-2-yl}piperidine-2,6-dione

[0473] Step Step

[0473] 1: Preparation 1: Preparation of 5-bromopentanal of 5-bromopentanal

DMSO DMSO oxalyl dichloride oxalyl dichloride Br Br m~-OH »BBr o OH TEA TEA

[0474] To a To

[0474] a solution solution of oxalyl of oxalyl dichloride dichloride (9.12 (9.12 g, 72 g, 72 mmol, mmol, 6 mL, 6 mL, 4.00 4.00 eq) eq) in in dichloromethane (50mL) dichloromethane (50 mL) was was added added a solution a solution of of dimethylsulfoxide dimethylsulfoxide (5.61 (5.61 g, 72 g, 72 mmol, mmol, 4.004.00 eq) eq)

in in dichloromethane (10 mL) dichloromethane (10 mL)atat-70 -70°C°Cover over3030min, min,andand then then 5-bromopentan-1-ol 5-bromopentan-1-ol (3.00 (3.00 g, 18 g, 18

mmol,1.00 mmol, 1.00eq) eq)was wasadded addedat atbelow below-60-60 °C.°C. TheThe resultingmixture resulting mixture waswas stirredatat-70 stirred -70°C°Cfor for11 hr. hr. Thin-layer chromatography(petroleum Thin-layer chromatography (petroleum ether:ethyl ether: ethylacetate acetate= =10:1) 10:1)showed showedthethe reactionwaswas reaction

complete. Triethylamine(14.54 complete. Triethylamine (14.54g,g, 144 144mmol, mmol,20 20 mL,mL, 8.00 8.00 eq) eq) waswas added added into into the the mixture mixture and and

225

2023248067 10 2023

reaction was the reaction the was stirred stirredatat-60 -60°C°Cfor for min.The 3030min. Themixture mixture was was poured into water poured into water (20 mL)and (20 mL) and stirred stirredfor for1 1min. min.The Theaqueous aqueous phase wasextracted phase was extracted with dichloromethane(20(20mLmL with dichloromethane x 3). x 3). TheThe

Oct combined organicphase combined organic phasewaswas washed washed withwith brine brine (20 (20 mL XmL 2), dried 2),x dried withwith anhydrous anhydrous sodium sodium

sulfate, sulfate, filtered andconcentrated filtered and concentrated in vacuum. in vacuum. The residue The residue was directly was directly usednext used for the forstep the next step without further without further purification. purification.5-bromopentanal (2.80 g, 5-bromopentanal (2.80 g, 17 17 mmol, 94% mmol, 94% yield)was yield) wasobtained obtained as as a a colorless oil. colorless oil.

[0475] StepStep

[0475] of 5-bromo-1,1-dimethoxypentane Preparation of 2: 2:Preparation 5-bromo-1,1-dimethoxypentane

trimethoxymethane trimethoxymethane O Br O methanol methanol Br Br O

[0476] To a To

[0476] a solution solution of 5-bromopentanal of 5-bromopentanal (2.80 (2.80 g, 16.97 g, 16.97 mmol,mmol, 1.00in eq) 1.00 eq) in methanol methanol (50 mL)(50 mL) was added was addedtrimethoxymethane trimethoxymethane (9.00 (9.00 g, 85 g, 85 mmol, mmol, 9 mL, 9 mL, 5.00 5.00 eq) 4-methylbenzenesulfonic eq) and and 4-methylbenzenesulfonic acid acid hydrate hydrate (161 mg, 0.85 (161 mg, 0.85 mmol, mmol,0.05 0.05eq)eq)atat2525°C. °C.The Theresulting resultingmixture mixturewas wasstirred stirred atat 25 25 °C °C for for 16 16 hr. hr.Thin-layer Thin-layer chromatography (petroleumether: chromatography (petroleum ether:ethyl ethylacetate acetate =10:1) showeda amajor =10:1) showed major new new

spot. spot. The The mixture waspoured mixture was pouredinto intowater water(40 (40mL) mL)andand stirredfor stirred for11 min. min. The Theaqueous aqueousphase phase waswas

extracted extracted with with ethyl ethyl acetate acetate (30 (30 mL x 3). mL X 3). The The combined organicphase combined organic phasewaswas washed washed withwith brine brine

(20 (20 mL 2), dried mL x x2), dried with anhydroussodium with anhydrous sodium sulfate,filtered sulfate, filtered and concentrated in and concentrated in vacuum. vacuum.The The residue was residue purified by was purified silica gel by silica gelcolumn column chromatography (petroleum chromatography (petroleum ether:ethyl ether: ethylacetate acetate =15:1). =15:1). 5-Bromo-1,1-dimethoxy-pentane 5-Bromo-1,1-dimethoxy-pentane (3.50 (3.50 g, 16.58 g, 16.58 mmol, mmol, 97% yield) 97% yield) was obtain was obtain as a colorless as a colorless oil. oil. H NMR ¹H NMR (400MHz, (400MHz, CDCl)CDC 4.37 ) (t,4.37 3 (t, J=5.6 J=5.6 Hz,3.41 Hz, 1H), 1H),(s,3.41 (s, 3.33 2H), 2H), (s, 3.336H), (s, 1.95 6H), 1.84 1.95 (m, - 1.84 (m, 2H), 1.67 -- 1.59 2H), 1.67 1.59 (m, (m, 2H), 1.54 - 1.45 2H), 1.54- 1.45 (m, (m, 2H). 2H).

[0477] Step Step

[0477] 3: Preparation 3: Preparation of (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]-2 of (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]=2-

phenyl-tetralin phenyl-tetralin

Bn Bn OH OH 0 0 Br" Br O

Cs 2CO DMF, CsCO, OC,1h1h 100100 3 , DMF, °C,

BnO Bn O\

[0478]

[0478] To To aa solution solution of of 4-[(1R,2S)-6-benzyloxy-2-phenyl-tetralin-1-yl]phenol 4-[(1R,2S)-6-benzyloxy-2-phenyl-tetralin-1-yl]phenol(500 (500 mg,mg, 1.231.23

mmol,1.00 mmol, 1.00eq) eq)inindimethylformamide dimethylformamide (5 mL) (5 mL) was added was added cesium cesium carbonate carbonate (1.2 mmol, (1.2 g, 3.69 g, 3.69 mmol, 3.00 eq) 3.00 eq) and and 5-bromo-1,1-dimethoxy-pentane 5-bromo-1,1-dimethoxy-pentane(390 (390 mg, mmol, mg, 1.84 1.84 mmol, 1.50 1.50 eq). Theeq). The was mixture mixture was stirred stirred at at100 100 °C °C for for1 1 hour. hour. The The reaction reaction mixture was diluted mixture was diluted with with water water(30 (30mL) mL)andand extracted extracted

226

Oct 2023 with ethyl with ethyl acetate acetate (15 (15 mL 2). The mL Xx 2). combinedorganic The combined organicphase phasewaswas washed washed withwith saturated saturated brine brine (15 (15

mLx x2), mL 2), dried dried with with anhydrous anhydroussodium sodium sulfate,filtered sulfate, filtered and andconcentrated concentratedininvacuum. vacuum.TheThe residue residue

was purified was purified by by silica silica gel gelcolumn chromatography(petroleum column chromatography (petroleum ether:ethyl ether: ethylacetate acetate= =50:1 50:1toto10:1) 10:1) to give (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin to give (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin (500 mg, (500 mg, 2023248067 10

0.93 mmol, 76% 0.93 mmol, yield) as 76% yield) as a awhite whitesolid. LC/MS solid. (ESI) LC/MS m/z:m/z: (ESI) 559.2 559.2 [M+23],

[M+23]*, 'H ¹H NMR (400MHz, NMR (400MHz,

CDCl 8 7.49 CDCl) 3)7.49 - 7.45 7.45 (m, 2H), (m, 2H), 7.38 -(m, 7.44 7.44 7.38 (m,7.37 2H), 7.31 -(m, 2H),- 7.37 7.311H), (m,7.21 7.13 -(m, 1H),- 7.21 7.133H), (m,6.90 3H), 6.90 -- 6.85 (m,2H), 6.85 (m, 2H),6.82 6.82 (dd, (dd, J=2.0, J=2.0, 7.2 7.2 Hz, 2H), Hz, 2H), 6.76J=2.4, 6.76 (dd, (dd, J=2.4, 8.4 Hz, 8.4 1H),Hz, 6.531H), 6.53 Hz, (d, J=8.8 (d, J=8.8 2H), Hz, 2H), 6.32 (d, J=8.8 6.32 (d, Hz, 2H), J=8.8 Hz, 2H),5.07 5.07(s, (s, 2H), 2H),4.38 4.38(t,(t, J=5.6 J=5.6Hz, Hz,1H), 1H), 4.25 4.25 (d,(d, J=4.8 J=4.8 Hz, Hz, 1H),1H), 3.843.84 (t, (t,

J=6.4 Hz, J=6.4 Hz, 2H), 2H), 3.41 - 3.28 3.41 3.28 (m,(m, 7H), 7H), 3.17 3.17 - 2.99(m, - 2.99 (m,2H), 2H),2.28 - 2.13 2.282.13 (m, (m, 1H),1H), 1.871.87 - 1.71 - 1.71 (m, (m, 3H),3H),

1.69 1.69 -1.60 1.60(m, (m,2H), 2H),1.54 1.54- -1.42 1.42(m, (m, 2H). 2H).

[0479] Step Step

[0479] 4: Preparation 4: Preparation of (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]- 2-phenyl of (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-

tetralin-6-ol tetralin-6-ol

Bn Bn Bn HO HO ó 0

- Pd/C, HH2 Pd/C, 0

MeOH, THF, 25 MeOH, THF, °C, 12 25C, h 12h o

[0480] To a To

[0480] a solution solution of (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl of (1R,2S)-6-benzyloxy-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-

tetralin (500 tetralin (500 mg, 0.93 mmol, mg, 0.93 mmol,1.00 1.00eq)eq) in in methanol methanol (20 (20 mL) mL) and tetrahydrofuran and tetrahydrofuran (20 mL)(20 wasmL) was added palladiumonon added palladium carbon carbon (200 (200 mg, mg, 10% purity) 10% purity) under under nitrogen nitrogen atmosphere. atmosphere. The suspension The suspension

was degassed was degassedand and purged purged with with hydrogen hydrogen 3 times. 3 times. The mixture The mixture was stirred was stirred under hydrogen under hydrogen (15 (15 psi) at psi) at 25 25 °C °C for 12 h. h. The reaction mixture The reaction mixture was wasfiltered filtered and and the the filter filter was was concentrated to give concentrated to give (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin-6-o (420 (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin-6-ol (420 mg, mg, crude) crude) as a as a white white

solid. LC/MS solid. (ESI)m/z: LC/MS (ESI) m/z:469.1 469.1[M+23].

[M+23]*.

[0481] Step Step

[0481] 5: Preparation 5: Preparation of 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl] of 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl

phenoxy]pentanal phenoxy]pentanal

HO Ho HO Ho

(2 M) H2 S04(2M) 0 HSO OO O THF70OC,5h THF, 70 °C, 0.5 h O O O O

227

[0482] To a To

[0482] a solution solution of (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin-6-ol of (1R,2S)-1-[4-(5,5-dimethoxypentoxy)phenyl]-2-phenyl-tetralin-6-ol

(420 mg, 0.94 (420 mg, 0.94 mmol, mmol,1.00 1.00eq)eq)inintetrahydrofuran tetrahydrofuran(75 (75mL) mL) waswas added added sulfuric sulfuric acidacid (2 in (2 M M water, in water, 18 mL,40.00 18 mL, 40.00eq). eq). TheThe mixture mixture was stirred was stirred at 70 at°C70 for°C 0.5for h. 0.5 Thinh.layer Thinchromatography layer chromatography (petroleum ether: ethyl (petroleum ether: ethyl acetate acetate == 3:1) 3:1)showed the reaction showed the reaction was completedand was completed anda anew newspot spotformed. formed. The reaction The reaction mixture mixturewas wasdiluted dilutedwith withwater water(40 (40mL)mL) andand extracted extracted withwith ethyl ethyl acetate acetate (20(20 mL mL x x

2). The 2). combined The combined organic organic phase phase was was washed washed with saturated with saturated sodium sodium bicarbonate bicarbonate (15 (15 mL) and mL) and saturated saturated brine (20 (20 mL mLx x2), 2),dried driedwith withanhydrous anhydrous sodium sodium sulfate, sulfate, filtered filtered andand concentrated concentrated in in

vacuum vacuum totogive give5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy] 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy] pentanal pentanal (370(370 mg, mg, 0.92 0.92

mmol,98% mmol, 98% yield)asasa awhite yield) whitesolid. solid.

[0483] Step Step

[0483] 6: Preparation 6: Preparation of tert-butyl of tert-butyl 4-(7-methoxy-1-oxo-1,3-dihydroisobenzofuran-5 4-(7-methoxy-1-oxo-1,3-dihydroisobenzofuran-5-

yl)piperazine-1-carboxylate yl)piperazine-1-carboxylate

NH NH 0 O O N 0 0 Boc'N Boc O O O DIPEA, NMP, 90 °C N N F O DIPEA, NMP, 90 °C N BocA,) Boc

[0484] To a To

[0484] a mixture mixture of 5-fluoro-7-methoxy-3H-isobenzofuran-1-one of 5-fluoro-7-methoxy-3H-isobenzofuran-1-one (1 mmol, (1 g, 5.49 g, 5.491 mmol, eq) 1 eq) and tert-butyl piperazine-1-carboxylate and tert-butyl piperazine-1-carboxylate (2.05 (2.05 g, g, 10.98 10.98 mmol, mmol, 22 eq) eq) in in 1-methylpyrrolidin-2-one 1-methylpyrrolidin-2-one

(6 (6 mL) wasadded mL) was addedN-ethyl-N-isopropylpropan-2-amine N-ethyl-N-isopropylpropan-2-amine (2.84(2.84 g, 21.96 g, 21.96 mmol,mmol, 3.834 mL, 3.83 mL, 4 eq) in eq) in

one portion. The one portion. mixture was The mixture wasstirred stirred at at 100 °C for 12 °C for 12 hours. hours. TLC (ethyl acetate/petroleum TLC (ethyl acetate/petroleum ether ether 1/1, Rf= = 1/1, = 0.1)indicated Rf=0.1) indicatedaa new newspot spotformed. formed.The The reactionmixture reaction mixture was was diluted diluted with with water water (20(20

mL)and mL) andextracted extractedwith withethyl ethylacetate acetate (40 (40 mL mLX x2). 2). The Thecombined combined organic organic layers layers were were washed washed

with water with water (15 (15 mL), mL), dried dried over oversodium sodiumsulfate, sulfate, filtered filtered and and concentrated concentrated under reducedpressure. under reduced pressure. The residue The residue was waspurified purified by by silica silica gel gel chromatography (petroleumether/ethyl chromatography (petroleum ether/ethylacetate acetate ==10/1 10/1 to to 1:1). 1:1). Tert-butyl Tert-butyl4-(7-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1-carboxylate 4-(7-methoxy-1-oxo-3H-isobenzofiuran-5-yl)piperazine-1-carboxylate (1 (1 g, g,2.87 2.87 mmol,52% mmol, 52% yield)was yield) was obtained obtained as as a yellow a yellow solid.LC/MS solid. LC/MS (ESI) (ESI) m/z:m/z: 349.3 349.3 [M+1]*;

[M+1]; ¹H NMR 'H NMR (400MHz, CDC 6.38 (400MHz, CDCl) 3) 6.38 (s, 1H), (s, 1H), 6.30 6.30 (s, 1H), (s, 1H), 5.135.13 (s, (s, 2H), 2H), 3.99 3.99 (s,(s,3H), 3H),3.62-3.59 3.62-3.59(m,(in, 4H), 4H),

3.42-3.35 (in,4H), 3.42-3.35 (m, 4H), 1.481.48 (s, (s, 9H).9H).

[0485] Step Step

[0485] 7: Preparation 7: Preparation of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(hydroxymethyl) of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(hydroxymethyl)-

6-methoxybenzoicacid 6-methoxybenzoic acid

228

0 0 0 0

NaOH NaOH OH OH O N -- OH N MeOH/THF H 2 0, MeOH/THF HO, OH N N N N BcN N Boc Boc Boc Boc

[0486] To a To

[0486] a mixture mixture of tert-butyl of tert-butyl 4-(7-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1 4-(7-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1

carboxylate (1 g, carboxylate (1 g, 2.87 2.87 mmol, mmol, 1 1eq) eq)ininmethyl methylalcohol alcohol(10 (10mL) mL) andand tetrahydrofuran tetrahydrofuran (10 (10 mL) mL) was was

added the solution added the solution of ofsodium hydroxide (459 sodium hydroxide (459 mg, 11.48 mmol, mg, 11.48 mmol, 44 eq) eq) in in water water (2 (2 mL). The mL). The

mixturewaswas mixture stirred stirred at at 20 20 °C for °C for 1 h.1 TLC h. (ethyl TLC (ethyl acetate/petroleum acetate/petroleum ether ether = 1/1, Rf = 1/1,indicated = 0) Rf= 0)aindicated a newspot new spotformed. formed.TheThe reaction reaction mixture mixture was concentrated was concentrated under reduced under reduced pressure pressure to removeto remove solvent. solvent. The The residue was diluted with water was diluted (20 mL) water (20 mL)and andextracted extractedwith withethyl ethylacetate acetate (30 (30 mL mLX x2). 2). The aqueous The aqueousphase phase was was adjusted adjusted to pHtovalue pH value to 4 ~to5 with 4 ~ 5hydrochloric with hydrochloric acid (1.5acid N), (1.5 then N), then filtered filtered and the solid and the solid was wascollected. collected.TheThe solid solid was was used used fornext for the the step nextwithout step without further further

purification. 4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-6-methoxy-benzoic purification. 4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-6-methoxy-benzoic acid acid (700 mg,1.68 (700 mg, 1.68mmol, mmol,58%58% yield, yield, 88% 88% purity) purity) was obtained was obtained as a white as a white solid.solid. LC/MSLC/MS (ESI) (ESI)

m/z: 367.3 m/z: 367.3[M+1].

[M+1].

[0487] Step Step

[0487] 8: Preparation 8: Preparation of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl) of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl) -2-formyl-6 -2-formyl-6-

methoxybenzoicacid methoxybenzoic acid

0 00 0 O o OH OH MnO 2 OH OH MnO Nit OH OH O N MeOH,6060°C°C MeOH, NN

Boc' N Boc'N Boc N Boc

[0488] To To

[0488] a mixture a mixture of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-6 of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-6-

methoxy-benzoic acid (650 methoxy-benzoic acid (650 mg, 1.77 mmol, mg, 1.77 mmol, 1 1eq) eq) and and in in methyl methyl alcohol alcohol (20 (20 mL) mL) was added was added

manganesedioxide manganese dioxide (1.54 (1.54 g, g, 17.74 17.74 mmol, mmol, 10ineq) 10 eq) oneinportion one portion at 20 at 20 °C °C nitrogen. under under nitrogen. The The mixture was mixture stirred atat5050°C°C for was stirred for 12 12hours. hours.LC/MS showed the LC/MS showed the reaction reaction was completed and was completed and desired product was formed. product was formed.The Thereaction reactionmixture mixturewaswas filteredand filtered andthe thesolution solutionwas wasconcentrated concentrated under vacuum. under vacuum.TheThe reaction reaction waswas usedused for the for the nextnext step step without without further further purification. purification. 4-(4-Tert 4-(4-Tert-

butoxycarbonylpiperazin-1-yl)-2-formyl-6-methoxy-benzoic butoxycarbonylpiperazin-1-yl)-2-formyl-6-methoxy-benzoicacid acid(600 (600mg, mg, 1.65 1.65 mmol, 92% mmol, 92%

yield) was yield) obtained as was obtained as aa yellow solid. LC/MS yellow solid. (ESI)m/z: LC/MS (ESI) m/z:365.3 365.3[M+1].

[M+1]'.

229

[0489] StepStep

[0489] 9: Preparation 9: Preparation of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(((2,6 of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(2,6-

dioxopiperidin-3-yl)amino)methyl)-6-methoxybenzoicacid dioxopiperidin-3-yl)amino)methyl)-6-methoxybenzoicacid

0 HN o 0 0 O O 2 H2N O -NHO NH OH OH HCI HCI NRoOH OH 0 o 0 H N O N N NaOAc, MeOH, NaBH NaOAc, MeOH, 3 CN NaBHCN N N NH NH N Boc'N Boc Boc'N,N Boc O 2023248067

[0490] To a To

[0490] a mixture mixture of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-formyl-6-methoxy-benzoic of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-formyl-6-methoxy-benzoiq

acid (600 (600 mg, 1.65 mmol, mg, 1.65 mmol,1 1eq) eq)and and3-aminopiperidine-2,6-dione 3-aminopiperidine-2,6-dione (407 (407 2.472.47 mg,mg, mmol, mmol, 1.5 eq, 1.5 eq,

HCl) in methyl HCI) in methylalcohol alcohol (10 (10 mL) mL)was was added added sodium sodium acetate acetate (203(203 mg, mg, 2.472.47 mmol, mmol, 1.5 and 1.5 eq) eq) and sodium cyanoborohydride sodium cyanoborohydride (310 (310 mg,mg, 4.944.94 mmol, mmol, 3 eq)3 in eq)one in one portion portion at °C. at 20 20 °C. The The mixture mixture was was

stirred stirredatat2020°C °Cfor for2 2h. h. LC/MS showedthe LC/MS showed thereaction reaction was wascompleted completedandand desired desired product product waswas

formed. Thereaction formed. The reaction mixture mixturewas wasconcentrated concentratedunder under vacuum. vacuum. The The residue residue was was purified purified by by

reverse reverse phase flash silica phase flash silicagel gelchromatography (120 gg SepaFlash chromatography (120 SepaFlashsilica silicagel gel column, column,eluent eluentofof 0-60% acetonitrileininwater 0~60% acetonitrile waterwith witha aflow flowrate rate of of 30 30 mL/min). mL/min).4-(4-Tert-butoxycarbonylpiperazin- 4-(4-Tert-butoxycarbonylpiperazin 1-yl)-2-[[(2,6-dioxo-3-piperidyl)amino]methyl]-6-methoxy-benzoic 1-yl)-2-[[(2,6-dioxo-3-piperidyl)amino]methyl]-6-methoxy-benzoic acidacid (300(300 mg, mg, 0.630.63 mmol, mmol,

38% yield) was 38% yield) wasobtained obtainedasasa awhite whitesolid. solid. LC/MS LC/MS (ESI) (ESI) m/z:477.4 m/z: 477.4 [M+1]'.

[M+1].

[0491] Step Step

[0491] 10: Preparation 10: Preparation of tert-butyl of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1 4-(2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1-

oxoisoindolin-5-yl)piperazine-1-carboxylate oxoisoindolin-5-yl)piperazine-1-carboxylate

O 0 00 O o o 00 NH PH OH O EDCI, TEA EDCI, TEA N O N o N N N NH HOBT, DCM DCM N N NH HOBT, N N N BocN N 0 BocNN Boc O Boc

[0492] To a To

[0492] a mixture mixture of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-[[(2,6-dioxo-3 of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-[[(2,6-dioxo-3-

piperidyl)amino]methyl]-6-methoxy-benzoic piperidyl)amino]methyl]-6-methoxy-benzoic acidacid (300(300 mg, mg, 0.63 0.63 mmol, mmol, 1 eq) 1ineq) in dichloromethane dichloromethane

(10 (10 mL) wasadded mL) was added N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide N-ethyl-N'-(3-dimethylaminopropyl)carbodiinide hydrochloride hydrochloride (181 mg, (181 mg,

0.94 0.94 mmol, 1.5 eq), mmol, 1.5 eq), N-hydroxybenzotrizole N-hydroxybenzotrizole(128 (128 mg,mg, 0.94 0.94 mmol, mmol, 1.5 eq), 1.5 eq), and and triethylamine triethylamine (191(191

mg, 1.89 mg, 1.89 mmol, mmol,3 3eq). eq).The Themixture mixturewaswas stirredatat20 stirred 20°C°Cfor for11 h. h. LC/MS LC/MS showed showed the the reaction reaction waswas

completed anddesired completed and desiredproduct productwas wasformed. formed. TheThe reaction reaction mixture mixture waswas quenched quenched by addition by addition of of water (15 mL), water (15 mL), and andthen thenextracted extracted with with dichloromethane dichloromethane(40(40 mLmL x 2). X 2). TheThe combined combined organic organic

layers layers were washedwith were washed withbrine brine(10 (10 mL), mL),dried driedover oversodium sodium sulfate,filtered sulfate, filtered and and concentrated concentrated under reduced under reduced pressure. pressure. The Theresidue residue was waspurified purifiedby bypreparative preparative TLC TLC (dichloromethane: (dichloromethane: methyl methyl

230

Oct 2023 10:1, Rf == 0.60). alcohol== 10:1, alcohol 0.60). Tert-butyl Tert-butyl4-[2-(2,6-dioxo-3-piperidyl)-7-methoxy-1-oxo-isoindolin 4-[2-(2,6-dioxo-3-piperidyl)-7-methoxy-1-oxo-isoindolin-

5-yl]piperazine-1-carboxylate (260 5-yl]piperazine-1-carboxylate (260mg, mg,0.57 0.57mmol, mmol,90%90% yield) yield) waswas obtained obtained as aaswhite a white solid. solid.

LC/MS (ESI) m/z: LC/MS (ESI) m/z: 459.4 [M+1]*.

[M+1]

[0493] Step Step

[0493] 11: Preparation 11: Preparation of 3-(7-methoxy-1-oxo-5-(piperazin-1-yl)isoindolin-2-yl) of 3-(7-methoxy-1-oxo-5-(piperazin-1-yl)isoindolin-2-yl) 2023248067 10

piperidine-2,6-dione piperidine-2,6-dione

0 O 00 O NH HCI/dioxane 0 00 NH N N O HCI/dioxane N O N O ND-Z N 7 20 OC 20 °C -N N Boc'N N HN HN Boc

[0494] To a To

[0494] a mixture mixture of tert-butyl of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-7-methoxy-1-oxo-isoindolin 4-[2-(2,6-dioxo-3-piperidyl)-7-methoxy-1-oxo-isoindolin-

5-yl]piperazine-1-carboxylate (300 (300 mg, mg,0.65 0.65mmol, mmol, 1 eq) 1 eq) inindioxane dioxane (10mL)mL) (10 waswas added added

hydrogen chloride/dioxane(4(4M,M,1717mL,mL, hydrogen chloride/dioxane 105.81 105.81 eq) eq) in in oneone portion. portion. The The mixture mixture waswas stirred stirred at at

20 °C 20 °C for for 22 h. h. The The reaction reaction mixture was concentrated mixture was concentrated under underreduced reducedpressure pressuretotoremove remove solvent. solvent.

The residue was The residue wasused usedfor for the the next next step step without further purification. without further purification.3-(7-Methoxy-1-oxo-5 3-(7-Methoxy-1-oxo-5-

piperazin-1-yl-isoindolin-2-yl)piperidine-2,6-dione (216 piperazin-1-yl-isoindolin-2-yl)piperidine-2,6-dione mg, 0.55 (216 mg, 0.55 mmol, mmol,83%83% yield, yield, HCIHCl salt) salt)

was obtained asasa awhite was obtained whitesolid. LC/MS solid. (ESI) LC/MS m/z:m/z: (ESI) 359.2359.2

[M+1]*; 1H-NMR

[M+1]; (400MHz, H-NMR MeOD) MeOD) (400MHz, 6: :

6.72 6.72 (s, 1H),6.60 (s, 1H), 1H),5.08-5.04 6.60(s,(s,1H), 5.08-5.04 (m, (m, 1H), 1H), 4.36-4.35 4.36-4.35 (m, (m, 2H), 2H), 3.92 (s, 3.92 (s, 3H), 3.66-3.65 3H), 3.66-3.65 (m, 5H), (m, 5H), 3.38-3.35 (m, 4H), 3.38-3.35 (m, 4H), 2.89-2.78 2.89-2.78 (m, (m, 1H), 1H), 2.77-2.67 2.77-2.67 (m, (m,1H), 1H),2.45-2.42 2.45-2.42(m, (m,1H), 1H),2.14-2.14 2.14-2.14(m, (m,1H). 1H).

[0495] StepStep

[0495] 12: Preparation 12: Preparation of 3-{5-[4-(5-{4-[(1R,2S)-6-hydroxy-2-phenyl-1,2,3,4 of 3-{5-[4-(5-{4-[(1R,2S)-6-hydroxy-2-phenyl-1,2,3,4-

tetrahydronaphthalen-1-yl]phenoxy}pentyl)piperazin-1-yl]-7-methoxy-1-oxo-2,3-dihydro-1H etrahydronaphthalen-1-yI]phenoxy}pentyl)piperazin-1-yl]-7-methoxy-1-oxo-2,3-dihydro-1H-

isoindol-2-yl}piperidine-2,6-dione (Exemplary isoindol-2-yl}piperidine-2,6-dione (Exemplary Compound Compound 2) 2)

0 0 0 0 NH NH I N N O N N O HO HO O( N -7O HNN N K-HN) HOHO HN HO HO O N N N N

NaBH 3CN,NaOAc, NaBHCN, NaOAc, AcOH, AcOH, MeOH, MeOH, DCMDCM

[0496] To a To

[0496] a mixture mixture of 3-(7-methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2-yl)piperidine-2,6 of 3-(7-methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2-yl)piperidine-2,6-

dione hydrochloride (89 dione hydrochloride (89mg, mg,0.23 0.23mmol) mmol) in methyl in methyl alcohol alcohol (5 mL) (5 mL) and dichloromethane and dichloromethane (1 mL) (1 mL)

was addedsodium was added sodium acetate(102 acetate (102 mg,mg, 1.251.25 mmol, mmol, 5 eq)5 in eq)one in portion one portion at 20at°C. 20 The °C. mixture The mixture was was stirred stirred at at20 20 °C for1 1 h, °C for h, then 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentanal 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentanal

231

Oct 2023 (100 mg,0.25 (100 mg, 0.25mmol, mmol, 1 was 1 eq) eq) added was added to the to the reaction reaction mixture mixture and for and stirred stirred 1 h. for 1 h. Sodium Sodium

cyanoborohydride (31mg,mg, cyanoborohydride (31 0.50 0.50 mmol, mmol, 2 eq) 2 eq) and and acetic acetic acidacid (0.05 (0.05 mL)mL) was was addedadded to reaction to the the reaction mixture. The mixture. Theresulting resulting solution solution was wasstirred stirred atat 20 20°C°Cforfor5 5h.h.LC/MS LC/MS showed showed the reaction the reaction was was completed and desired completed and desired product product was was formed. formed. The Thereaction reaction mixture mixturewas wasconcentrated concentratedunder under 2023248067 10

reduced pressure reduced pressure toto remove remove solvent. solvent. TheThe residue residue was was purified purified by preparative by preparative HPLC (column: HPLC (column:

PhenomenexSynergi Phenomenex SynergiC18 C18150150 x 25 X 25 x 10 X 10 um;um; mobile mobile phase: phase: [water(0.05%HC)-acetonitrile];

[water(0.05%HCl)-acetonitrile];

B%: 35%-55%, B%: 35%-55%, 7.8 3-[5-[4-[5-[4-[(1R,2S)-6-Hydroxy-2-phenyl-tetralin-1- 7.8 min). min). 3-[5-[4-[5-[4-[(1R,2S)-6-Hydroxy-2-phenyl-tetralin-1 yl]phenoxy]pentyl]piperazin-1-yl]-7-methoxy-1-oxo-isoindolin-2-yl]piperidine-2,6-dione(109.9 yl]phenoxy]pentyl]piperazin-1-yl]-7-methoxy-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (109.9 mg, 0.14 mg, 0.14 mmol, mmol,56%56% yield, yield, 100% 100% purity, purity, HCI HCl salt)salt) was was obtained obtained as a as a white white solid. solid. LC/MSLC/MS (ESI) (ESI) m/z: 743.7 m/z: 743.7 [M+1>]; 'H-NMR(400MHz,

[M+1]; ¹H-NMR (400MHz, DMSO-d6) DMSO-d6) 8 10.93 10.93 (s, 1H), (s, 1H), 10.56-10.43 10.56-10.43 (m,(in, 1H), 1H), 9.18 9.18-

9.13 (m, 9.13 (in, 1H), 1H), 7.16-7.13 7.16-7.13 (m, (in,3H), 3H),6.84-6.83 6.84-6.83(d,(d,JJ= 6.4Hz,2H), = 6.4Hz, 2H),6.69 6.69(s,(s,1H), 1H),6.62-6.61 6.62-6.61(m,(in, 2H), 2H),

6.55-6.52 (in, 3H), 6.55-6.52 (m, 3H), 6.28-6.26 6.28-6.26 (d, (d, J= J = 8.4Hz, 8.4Hz, 2H), 4.99-4.97 (m, 2H), 4.99-4.97 (in, 1H), 1H), 4.29-4.25 4.29-4.25 (m, (in,1H), 1H),4.23-4.18 4.23-4.18 (in, 1H), 4.17-4.15 (m, 1H), 4.17-4.15 (m, (in,1H), 1H), 4.06-4.00 4.06-4.00(m, (in,2H), 2H),3.85-3.83 3.85-3.83(m,(in, 5H), 5H), 3.56-3.53 3.56-3.53 (m,(in, 1H), 1H), 3.34-3.33 3.34-3.33

(in, 4H), 3.10-3.02 (m, 4H), 3.10-3.02 (m, (in, 4H), 4H), 3.00-2.85 3.00-2.85 (m, (in,2H), 2H),2.60-2.58 2.60-2.58(m,(in, 3H), 3H), 2.16-2.08 2.16-2.08 (m,(in, 1H), 1H), 1.91-1.88 1.91-1.88

(in, 1H), 1.76-1.69 (m, 1H), 1.76-1.69 (m, (in, 5H), 5H), 1.43-1.41 1.43-1.41 (m, (in,2H). 2H).

[0497] Exemplary

[0497] Exemplary Synthesisof ofExemplary Synthesis Exemplary Compound Compound 3: 3-[5-[4-[5-[4-[(1R,2S)-6 3: 3-[5-[4-|5-[4-[(1R,2S)-6- hydroxy-2-phenyl-tetralin-1-yl] hydroxy-2-phenyl-tetralin-1-yl| phenoxy]pentyl]piperazin-1-yl]-4-methoxy-1-oxo phenoxy|pentyl|piperazin-1-yl]-4-methoxy-1-oxo-

isoindolin-2-yl]piperidine-2,6-dione isoindolin-2-yl|piperidine-2,6-dione

[0498] Step Step

[0498] 1: Preparation 1: Preparation of 5-bromo-4-iodo-3H-isobenzofuran-1-one of 5-bromo-4-iodo-3H-isobenzofuran-1-one

0 0 o O NIS NIS

trifluoromethanesulfonic acid Br Br Br trifluoromethanesulfonic acid Br

[0499] To a To

[0499] a solution solution of 5-bromo-3H-isobenzofuran-1-one of 5-bromo-3H-isobenzofuran-1-one (50 g, 234.71 (50 g, 234.71 mmol, 1 mmol, eq) in 1 eq) in trifluoromethanesulfonic acid trifluoromethanesulfonic acid (680 (680 g, g, 4.53 4.53 mol, 400 mL, mol, 400 mL,19.30 19.30eq) eq)was wasadded added 1-iodopyrrolidine 1-iodopyrrolidine-

2,5-dione (55.45 2,5-dione (55.45 g, g, 246.45 mmol,1.05 246.45 mmol, 1.05eq) eq)atat 00 °C °C inin portions. portions. The mixture was The mixture wasallowed allowedtotowarm warm to 15 to 15 °C °C and held for and held for 16 16 h. h. TLC (petroleumether: TLC (petroleum ether: ethyl ethyl acetate acetate == 5:1) 5:1) showed nostarting showed no starting material remained material andtwo remained and twonew new spots(R=0.4,0.5) spots (Rf= 0.4, formed. 0.5) formed. The reaction The reaction mixture mixture was poured was poured

into ice-water (1 L) and yellow solid precipitated. The mixture was filtered and the filter cake into ice-water (1L) and yellow solid precipitated. The mixture was filtered and the filter cake

was washed was washedwith withwater. water.The Thefilter filter cake cake was wasdissolved dissolvedininethyl ethyl acetate acetate (500 (500 mL) mL)and andthe theresulting resulting orange solution orange solution waswas dried dried over over sodium sodium sulfate.sulfate. The was The mixture mixture wasandfiltered filtered and the the filtrate wasfiltrate was

concentrated to afford a yellow solid. The residue was triturated with ethyl acetate (50 mL), concentrated to afford a yellow solid. The residue was triturated with ethyl acetate (50 mL),

232 filtered filteredand andwashed ethyl acetate with ethyl acetate (10 (10 mL mL Xx 2). 5-Bromo-4-iodo-3H-isobenzofuran-1-one (40 (40 2). 5-Bromo-4-iodo-3H-isobenzofuran-1-one 2023248067 10 Oct 2023 washed with g, 118.02 mmol, g, 118.02 50% mmol, 50% was yield)was yield) obtained obtained as as a yellow a yellow solid.¹H'HNMR solid. NMR (400MHz, (400MHz, 7.83 3 )8 7.83 CDCl) CDCl

(d, (d, J= 8.0 Hz, J = 8.0 Hz,1H), 7.77 1H),7.77 (d,(d, J =J= 8.08.0 Hz, Hz, 5.10 5.10 1H),1H), (s, 2H). (s, 2H).

[0500] Step Step

[0500] 2: Preparation 2: Preparation of 5-bromo-4-hydroxy-3H-isobenzofuran-1-one of 5-bromo-4-hydroxy-3H-isobenzofuran-1-one

o O 0 NaOH Cu 2 0, NaOH CuO, o O Br Br H 20, DMA, HO, DMA,80°C 80 °C Br Br OH OH

[0501] To a To

[0501] a mixture mixture of 5-bromo-4-iodo-3H-isobenzofuran-1-one of 5-bromo-4-iodo-3H-isobenzofuran-1-one (40 g,mmol, (40 g, 118.02 118.02 mmol, 1 eq), 1 eq), sodium hydroxide(23.60 sodium hydroxide (23.60g, g,590.10 590.10 mmol, mmol, 5 eq) 5 eq) in water in water (400(400 mL) mL) and N,N-dimethylacetamide and N,N-dimethylacetamide

(200 mL)was (200 mL) wasadded added cuprous cuprous oxide oxide (3.38 (3.38 g, 23.60 g, 23.60 mmol, mmol, 2.40.2 2.4 mL, mL,eq). 0.2 The eq).reaction The reaction mixture mixture

was heated was heatedtoto8080°C°Candand held held for for 16 TLC 16 h. h. TLC (petroleum (petroleum ether ether : ethyl : ethyl acetate acetate = 1:1, 1:1,= Rf = Rf 0.3)= 0.3) showed thereaction showed the reactionwaswas completed. completed. The The reaction reaction mixture mixture was poured was poured into 1N into IN hydrochloride hydrochloride

solution solution (400 mL)and (400 mL) andextracted extractedwith withethyl ethylacetate acetate(400 (400mLmL x 2). X 2). TheThe combined combined organic organic layerslayers

were concentrated were concentratedandand dissolved dissolved in ethyl in ethyl acetate acetate (500(500 mL), mL), washedwashed with saturated with saturated aqueous aqueous sodium bicarbonate(150 sodium bicarbonate (150mL), mL), brine brine (150 (150 mL) mL) and then and then drieddried over over sodium sodium sulfate. sulfate. The mixture The mixture

filteredand wasfiltered was andthethe filtratewaswas filtrate concentrated concentrated to afford to afford a residue. a residue. The was The residue residue was with triturated triturated with ethyl acetate ethyl acetate(20 (20mL), mL), filtered filtered and and washed washed withacetate with ethyl ethyl acetate (10give (10 mL) to mL) to give a solid. Thea filtrate solid. The filtrate was further concentrated was further concentrated andand trituratedwith triturated with ethyl ethyl acetate. acetate. 5-Bromo-4-hydroxy-3H 5-Bromo-4-hydroxy-3H-

isobenzofuran-1-one(14.5 isobenzofuran-1-one (14.5g,g, 60.15 60.15mmol, mmol,50%50% yield, yield, 95%95% purity) purity) was was obtained obtained as a as a white white solid. solid.

'H NMR ¹H NMR (400MHz, (400MHz, DMSO)DMSO) 10.90 (s,10.90 1H), (s, 7.721H), (d, 7.72 J = 8.0 Hz, 8.0 (d, J= 1H),Hz, 7.23 (d, 7.23 1H), J = 8.0 Hz, 8.0 (d, J= 1H),Hz, 1H), 5.35 (s, 2H). 5.35 (s, 2H). 3: 3:

[0502] StepStep

[0502] Preparation of 5-bromo-4-methoxy-3H-isobenzofuran-1-one Preparation of 5-bromo-4-methoxy-3H-isobenzofuran-1-one

o O 0 o CH 3 1, KKCO CHI, 2 CO3 oO O O Br Br acetone acetone Br Br OH OH O1

[0503] To a To

[0503] a mixture mixture of 5-bromo-4-hydroxy-3H-isobenzofuran-1-one of 5-bromo-4-hydroxy-3H-isobenzofuran-1-one (3 mmol, (3 g, 13.10 g, 13.10 mmol, 1 eq) 1 eq) in acetone acetone (20 mL) wasadded mL) was addediodomethane iodomethane (17.5 (17.5 g, 123.29 g, 123.29 mmol, mmol, 7.7 9.41 7.7 mL, mL, 9.41 eq) and eq) and

potassiumcarbonate potassium carbonate(5.43 (5.43g,g, 39.30 39.30 mmol, mmol,3 3eq). eq).The Themixture mixture was was stirredatat2020°C°Cfor stirred for1515h.h. TLC TLC (ethyl acetate: petroleum (ethyl acetate: petroleum ether ether = 1:3, = 1:3, Rf =Rf= 0.37)0.37) indicated indicated reaction reaction was completed. was completed. The reaction The reaction

mixture was mixture wasquenched quenchedby by addition addition of of water(10(10mL), water mL), andand then then extracted extracted with with ethyl ethyl acetate(20(20 acetate

233

Oct 2023 2). The mLx x2). mL Thecombined combined organic organic layerswere layers were washed washed withwith saturated saturated sodium sodium bicarbonate bicarbonate (10X (10 mL mL x 2), dried 2), dried over over sodium sulfate, filtered sodium sulfate, filteredand andconcentrated concentratedunder underreduced pressure.5-Bromo-4 reduced pressure. 5-Bromo-4-

methoxy-3H-isobenzofuran-1-one methoxy-3H-isobenzofuran-1-one (2.9(2.9 g, 11.93 g, 11.93 mmol, mmol, 91% yield) 91% yield) was obtained was obtained as a yellow as a yellow solid.solid.

'H NMR ¹H NMR (400MHz, (400MHz, CDCl)CDC 7.72 ) (d,7.72 J = (d, 3 8.0J= Hz,8.0 Hz,7.49 1H), 1H),(d, 7.49 J =(d,8.0 J=Hz,8.01H), Hz, 5.44 1H), (s, 5.442H), (s, 2H), 2023248067 10

4.00 (s, 3H). 4.00 (s, 3H).

[0504] StepStep

[0504] 4: Preparation 4: Preparation of tert-butyl of tert-butyl 4-(4-methoxy-1-oxo-3H-isobenzofuran 4-(4-methoxy-1-oxo-3H-isobenzofuran -5-yl) -5-yl)

piperazine-1-carboxylate piperazine-1-carboxylate

ONH NH 0 o O N Boc'N Boc

O Br Br Pd(dba), XantPhos Pd 2 (dba) 3, XantPhos N N OI*_I potassium phosphate potassium phosphate Boc'NN O, dioxane, 100°C°C dioxane, 100 Boc

[0505]

[0505] A vial A vial was chargedwith was charged with5-bromo-4-methoxy-3H-isobenzofuran-1-on 5-bromo-4-methoxy-3H-isobenzofuran-1-one (500 mg,(500 2.06mg, 2.06 mmol,1 1eq), mmol, eq), tert-butyl tert-butyl piperazine-1-carboxylate piperazine-1-carboxylate (383 mg, 2.06 (383 mg, 2.06 mmol, mmol,1 1eq), eq), tris(dibenzylideneacetone)dipalladium(0) (188mg, tris(dibenzylideneacetone)dipalladium(0) (188 mg,0.20 0.20mmol, mmol, 0.10.1 eq),XantPhos eq), XantPhos (119(119 mg, mg, 0.20 0.20

mmol,0.1 mmol, 0.1 eq), eq), potassium potassiumphosphate phosphate(873 (873mg,mg, 4.11 4.11 mmol, mmol, 2 eq) 2 eq) and and dioxane dioxane (5 mL). (5 mL). The The mixture was mixture waspurged purgedwith withnitrogen nitrogenand andheated heatedtoto100100°C°C forfor 1616 h.h.TLC TLC (ethylacetate: (ethyl acetate:petroleum petroleum ether ether == 1:3) 1:3) showed reaction was showed reaction was complete. complete.The Themixture mixture was was diluted diluted with with ethylacetate ethyl acetate(30 (30mL) mL) and washedwith and washed withwater water(30 (30mL). mL).TheThe aqueous aqueous layer layer was was extracted extracted withwith ethyl ethyl acetate acetate (15(15 mL mL x 3). X 3).

The organic The organic layer layer was waswashed washedwith withbrine brine(30 (30mL) mL) andand dried dried over over sodium sodium sulfate. sulfate. TheThe crude crude was was

purified by silica gel chromatography (ethyl acetate : petroleum ether 1:20 to 1:6). Tert-butyl purified by silica gel chromatography (ethyl acetate : petroleum ether = 1:20 to=1:6). Tert-butyl

4-(4-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1-carboxylate 4-(4-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1-carboxylate (700 (700 mg, 2.01 mg, 2.01 mmol,mmol, 97% 97% yield) was yield) obtained as was obtained as aa yellow solid. LC/MS yellow solid. (ESI)m/z: LC/MS (ESI) m/z:349.2 349.2[M+1].

[M+1]*.

[0506] Step Step

[0506] 5: Preparation 5: Preparation of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2- of (4-(4-tert-butoxycarbonylpiperazin-1-yl)-2- (hydroxylmethyl) (hydroxylmethyl)-

3-methoxy-benzoicacid 3-methoxy-benzoic acid

0 0 o o NaOH NaOH OH OH O N OH N THF, 20 THF, HH2O OH N N Boc'N Boc N O Boc'N N O, Boc

[0507] To a To

[0507] a solution solution of tert-butyl of tert-butyl 4-(4-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1 4-(4-methoxy-1-oxo-3H-isobenzofuran-5-yl)piperazine-1-

carboxylate (700 mg, carboxylate (700 mg,2.01 2.01 mmol, mmol,1 eq) 1 eq)inintetrahydrofuran tetrahydrofuran(4(4mL) mL)andand water water (4 (4 mL)mL) was was added added

sodiumhydroxide sodium hydroxide(401 (401mg,mg, 10.05 10.05 mmol, mmol, 5 eq). 5 eq). The The mixture mixture was stirred was stirred at °C at 20 20 for °C for 16 h. 16 h. TLCTLC

234

(ethyl (ethyl acetate: acetate:petroleum petroleum ether ether== 1:2) 1:2)showed showed reaction reaction was complete. The Themixture mixturewas wasadjusted adjusted 2023248067 10 Oct 2023

was complete.

to to pH pH == 44 with with aqueous aqueoushydrochloric hydrochloricacid acid(1M) (1 M) andand extracted extracted with with ethyl ethyl acetate(10(10mlml acetate x 3).The x 3). The organic layer was organic layer washedwith was washed withbrine brine(20 (20mL) mL)andand dried dried over over sodium sodium sulfate.TheThe sulfate. crude crude material material

was not further was not further purified. purified.4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl) 4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-3- -3

methoxy-benzoic acid(700 methoxy-benzoic acid (700mg,mg, crude) crude) waswas obtained obtained as aasyellow a yellow solid. solid.

[0508] Step Step

[0508] 6: Preparation 6: Preparation of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2- of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2- formyl-3 formyl-3-

methoxybenzoic acid methoxybenzoic acid

O O 0 O OH MnO OH OH MnO2 OH N OH OH DCM N O O N DCM N Boc'N Boc N O O Boc'N Boc N 0, O

[0509]

[0509] To To aa solution solution of of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-3 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-(hydroxymethyl)-3-

methoxy-benzoic methoxy -benzoicacid acid(700 (700mg, mg,1.91 1.91mmol, mmol, 1 eq) 1 eq) in in dichloromethane dichloromethane (10 (10 mL) mL) was added was added

manganesedioxide manganese dioxide(2.49 (2.49g,g,28.66 28.66mmol, mmol,15 15 eq). eq). TheThe mixture mixture was was stirred stirred at at 2020 °C °C forfor 1 1 h.h.TLC TLC (dichloromethane: methanol= (dichloromethane: methanol 20:1)showed = 20:1) showed reaction reaction waswas complete. complete. The The mixture mixture was diluted was diluted

with dichloromethane(10 with dichloromethane (10mL) mL)andand filteredthrough filtered througha apad padofofCelite. Celite. The Thefiltrate filtrate was concentrated was concentrated

in vacuum. Thecrude vacuum. The crudeproduct productwas was purifiedbybysilica purified silicagel gel column columnchromatography chromatography (dichloromethane: methanol= (dichloromethane: methanol = 100:1to 100:1to 60:1).4-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2- 60:1). 4-(4-(Tert-butoxycarbonyl)piperazin-1-yl)-2 formyl-3-methoxybenzoic acid formyl-3-methoxybenzoic acid (300 (300 mg,mg, 0.82 0.82 mmol, mmol, 43% yield) 43% yield) was obtained was obtained as a yellow as a pale pale yellow solid. solid.

[0510] Step Step

[0510] 7: Preparation 7: Preparation of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2 of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2 -[[(2,6-dioxo -[(2,6-dic -3- -3 piperidyl)amino]methyl]-3-methoxy-benzoic piperidyl)amino]methyl]-3-methoxy-benzoic acid acid

0 0N0 o o NH O CIH HN o OH OH CIHH 2 N O OQHH 0 o H N N -OO NaOAc, NaBH NaOAc, 3CN, NaBHCN, N N N NH NH N O MeOH N o Boc'N Boc O, MeOH Boc'N Boc O0 O

[0511]

[0511] To To aa mixture mixture of of 3-aminopiperidine-2,6-dione 3-aminopiperidine-2,6-dione(135 (135mg,mg, 0.82 0.82 mmol, mmol, 1 eq, 1 eq, HCIHCl salt) salt) in in

methanol(2(2 mL) methanol mL)and and dichloromethane dichloromethane (4 mL) (4 mL) was was addedadded sodium sodium acetate acetate (2703.29 (270 mg, mg,mmol, 3.29 4mmol, 4 eq). Themixture eq). The mixturewaswas stirred stirred at°C at 20 20for °C10formin, 10 then min,4-(4-tert-butoxycarbonylpiperazin-1-yl) then 4-(4-tert-butoxycarbonylpiperazin-1-yl) -

2-formyl-3-methoxy-benzoic 2-formyl-3-methoxy-benzoic acid acid (300 (300 mg,mg, 0.820.82 mmol, mmol, 1 eq)1 was eq) added was added andmixture and the the mixture was was stirred stirredfor for10 10min. min.Sodium cyanoborohydride(103 Sodium cyanoborohydride (103mg,mg, 1.65 1.65 mmol, mmol, 2 eq) 2 eq) was was added added and the and the

235 mixture was wasfurther further stirred stirred for for40 40min. min. LCMS showed reaction waswas complete. The The mixture was 2023248067 10 Oct 2023 mixture LCMS showed reaction complete. mixture was adjusted adjusted to to pH 4-5 with pH == 4-5 with aqueous aqueoushydrochloric hydrochloricacid acidsolution solution(1(1 M) M)and andextracted extractedwith withethyl ethyl acetate acetate (10 (10 mL 3). The mL xx 3). organic layer The organic layer was was dried dried over over sodium sodiumsulfate. sulfate. The Thecrude crudeproduct productwas wasnot not further further purified. purified.4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-[[(2,6-dioxo-3 4-(4-Tert-butoxycarbonylpiperazin-1-yl)-2-|[(2,6-dioxo-3- piperidyl)amino]methyl]-3-methoxy-benzoic piperidyl)amino]methyl]-3-methoxy-benzoic acidacid (400(400 mg, mg, crude) crude) was obtained was obtained as a as a white white solid. solid.

LC/MS (ESI) m/z: LC/MS (ESI) m/z: 477.1 477.1 [M+1]*.

[M+1].

[0512] Step Step

[0512] 8: Preparation 8: Preparation of tert-butyl of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo 4-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo-

isoindolin-5-yl]piperazine-1-carboxylate isoindolin-5-yl]piperazine-1-carboxylate

0 o 00 o O NH NH OHH O HATU HATU N 0 A N O N N1N !NHNH DMF DIPEA, DMF DIPEA, N N N Boc'NN o O N Boc'N O, o Boc Boc O

[0513]

[0513] To of 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-[[(2,6-dioxo-3 solution of To aa solution 4-(4-tert-butoxycarbonylpiperazin-1-yl)-2-[(2,6-dioxo-3-

piperidyl)amino] methyl]-3-methoxy-benzoic piperidyl)amino] methyl]-3-methoxy-benzoicacidacid (400 (400 mg, mg, 0.840.84 mmol, mmol, 1 eq) 1in eq) in dimethylformamide dimethylformamide (5 (5 mL) mL) was was added added o-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium o-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium

hexafluorophosphate (383mg,mg, hexafluorophosphate (383 1.01mmol, 1.01 mmol, 1.2 1.2 eq).eq). TheThe solution solution waswas stirred stirred forfor 10min, 10min, then then N,N N,N-

diisopropylethylamine (325 disopropylethylamine (325 mg,mg, 2.52 2.52 mmol, mmol, 3 eq) 3 eq) was was added. added. The solution The solution was stirred was stirred at °C at 20 20 °C for for 20 20 min. min. LCMS showed LCMS showed reaction reaction was was complete. complete. The solution The solution was diluted was diluted with with ethylethyl acetate acetate

(40 mL) andwashed mL) and washed with with water water (30(30 mL mL 5) and X 5)x and brine brine (40 (40 mL). mL). The The organic organic layerlayer was was drieddried

over sodiumsulfate. over sodium sulfate. Tert-butyl Tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo-isoindolin-5-yl] 4-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo-isoindolin-5-jyl]

piperazine-1-carbo xylate piperazine-1-carbo xylate (400 (400 mg, mg,crude) crude)was wasobtained obtainedasasa apale paleyellow yellowsolid. solid. LC/MS LC/MS (ESI) (ESI)

m/z: 459.1 m/z: 459.1[M+1].

[M+1].

[0514] Step Step

[0514] 9: Preparation 9: Preparation of 3-(4-methoxy-1-oxo-5-piperazin-1-yl-isoindolin of 3-(4-methoxy-1-oxo-5-piperazin-1-yl-isoindolin -2-yl)-2-yl)

piperidine-2,6-dione piperidine-2,6-dione

o o O NH HCl/dioxane NH N o N o N N NO HCI/dioxane dioxane N Ndioxane NN0 N Boc'N Boc N o O, HN O, o HCI HCI

[0515] To a To

[0515] a mixture mixture of tert-butyl of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo-isoindolin 14-[2-(2,6-dioxo-3-piperidyl)-4-methoxy-1-oxo-isoindolin-

5-yl] 5-yl] piperazine-1-carboxylate (400 mg, piperazine-1-carboxylate (400 mg, 0.87 0.87 mmol, mmol,1 eq) 1 eq)inindioxane dioxane(2(2mL) mL)waswas added added

hydrochloric acid in hydrochloric acid in dioxane (4 M, dioxane (4 M, 44 mL, mL,18.34 18.34eq). eq). The Themixture mixturewas was stirredatat20 stirred 20 °C °Cfor for 10 10 min min

236

Oct 2023 and was removed solvent was and solvent removed under under vacuum. vacuum. 3-(4-Methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2 -(4-Methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2

yl)piperidine-2,6-dione (350 yl)piperidine-2,6-dione (350 mg, mg, crude, HCl crude, HCI salt) was salt) was obtained obtainedasas aa white solid. LC/MS white solid. (ESI) LC/MS (ESI)

m/z: 359.1 m/z: 359.1[M+1].

[M+1].

[0516] Step Step

[0516] 10: Preparation 10: Preparation of 3-[5-[4-[5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl] of 3-[5-[4-[5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl] 2023248067 10

phenoxy]pentyl]piperazin-1-yl]-4-methoxy-1-oxo-isoindolin-2-yl]piperidine phenoxylpentyl]piperazin-1-yl]-4-methoxy-1-oxo-isoindolin-2-yllpiperidine -2,6-dione -2,6-dione

(Exemplary (Exemplary Compound 3) Compound 3) HO HO o H IZ H o HN O O N O o HN NN N N N

0 Np o / O N o N N O AcONa, NaBH 3CN, MeOH N N O AcONa, NaBHCN, MeOH HCI HCI NH NH NH O

0 HO Ho O

[0517] To a To

[0517] a mixture mixture of 3-(4-methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2-yl)piperidine-2,6 of 3-(4-methoxy-1-oxo-5-piperazin-1-yl-isoindolin-2-yl)piperidine-2,6-

dione (100 mg, dione (100 mg,0.25 0.25mmol, mmol, 1 eq,HCI 1 eq, HCl salt)inindichloromethane salt) dichloromethane(4 (4 mL)mL) and and methanol methanol (1 was (1 mL) mL) was added sodiumacetate added sodium acetate(83(83mg,mg, 1.01 1.01 mmol, mmol, 4 eq). 4 eq). The mixture The mixture was stirred was stirred at 20 at °C20 for°C10for 10 min. min.

Then 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentanal Then 5-[4-[(1R,2S)-6-hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentanal (101 0.25 (101 mg, mg,mmol, 0.25 mmol, 1.00 1.00 eq) was addedand was added andthethemixture mixture waswas stirred stirred forfor 10 10 min. min. Sodium Sodium cyanoborohydride cyanoborohydride (31 mg,(31 mg,

0.51 mmol, mmol,2 2eq) eq)was was added added to the to the mixture mixture and and stirring stirring was was keptkept for min. for 40 40 min. LCMS LCMS and TLC and TLC

(dichloromethane: methanol= (dichloromethane: methanol = 10:1) 10:1) showed showed reaction reaction was was complete. complete. Solvent Solvent was removed was removed under under vacuum. Thecrude vacuum. The crudeproduct product was was purified purified by by prep-TLC prep-TLC (dichloromethane: (dichloromethane: methanol methanol = 10:1). = 10:1). 3-[5- 3-[5

[4-[5-[4-[(1R,2S)-6-Hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentyl]piperazin-l-yl]-4-methoxy

[4-[5-[4-[(IR,2S)-6-Hydroxy-2-phenyl-tetralin-1-yl]phenoxy]pentyl]piperazin-1-yl]-4-nethoxya

1-oxo-isoindolin-2-yl]piperidine-2,6-dione (55 1-oxo-isoindolin-2-yl]piperidine-2,6-dione (55 mg, mg, 0.07 0.07 mmol, mmol, 29% yield, 29% yield, 99% was 99% purity) purity) was obtained obtained as as aawhite whitesolid. LC/MS solid. LC/MS(ESI) (ESI)m/z: 743.3 m/z: [M+1]*; 743.3 [M+1]; 'H-NMR (400MHz,DMSO-d6) ¹H-NMR (400MHz, DMSO-d6) 8 10.96 (s, (s, 1H), 1H), 9.12 9.12 (s, (s, 1H), 1H), 7.39 7.39 (d, (d,J=8.0 J=8.0Hz, Hz, 1H), 7.25 -- 6.98 1H), 7.25 6.98 (m, (m, 4H), 4H), 6.83 6.83 (d, (d,J=6.8 J=6.8 Hz, Hz, 2H), 2H),

6.72 -- 6.43 6.43 (m, 5H), 5H), 6.26 6.26 (d, (d, J=8.6 Hz, 2H), J=8.6 Hz, 2H), 5.06 5.06 (dd, (dd, J=5.0, J=5.0, 13.2 13.2 Hz, Hz, 1H), 1H),4.56 4.56- -4.11 4.11 (m, (m,3H), 3H), 3.94 -- 3.70 3.70 (m, 3.30 -- 3.25 (m, 5H), 3.30 3.25 (m, (m, 1H), 3.21 -- 2.77 1H), 3.21 2.77 (m, (m, 8H), 8H), 2.64-2.55 (m, 5H), 2.64-2.55 (m, 2.46 -- 2.26 5H), 2.46 2.26 (m, (m, 2H), 2.16- -1.94 2H), 2.16 1.94(m,(m, 2H), 2H), 1.801.80 - 1.22 - - 1.22 (m,(m, 7H). 7H).

237

[0518] B. B.

[0518] Exemplary Exemplary Synthetic Synthetic Schemes Schemes for Exemplary for Exemplary Androgen Androgen Receptor Receptor Binding Binding

Moiety Based Compounds Moiety Based Compounds

[0519]

[0519] General Synthetic Scheme General Synthetic B-i Scheme B-1

0 N "OHOH Nr OH OH O HN N Dess-Martin Dess-Martin N NN DIEA, DMF, DIEA, 100°CC DMF, 100 o O DCM, rt, overnight DCM, rt, overnight 0 o FF r 0 o 0 o ZI 2023248067 H HN o N HN N O o o o TFHN TFA N O'0O N o H N NH N NH N H N N o H 4N HCI 4NHCI o 0 - 0 O Dioxane N NaBH(OAc) 3 , DOM, NaBH(OAc), DCM, rt rt CNNN N o N N o N N o \40 -aHOHo

HCI HCI O NC NC O "NH 2 CI 0NC0N NH O o CI o o O NC IZ NH CI N o HATU,DIEA, HATU, DMF DIEA,DMF CI N N C:4O - N N N o

[0520]

[0520] General Synthetic Scheme General Synthetic B-2 Scheme B-2

0 o 0 o BocN Boc 0 N o Br Br Br Br CKH0OxNen Br Br - OH MH7 OH20 0 RuPhosPdCs2CO 3 N RuPhosPd, CsCO 0 KMnO, NaOH HSO N O CNN H2 0/dioxane OH OHMeOH, 700,2020hh MeOH, 70°C, O 0 P~,rfu,1 PhMe, reflux, 16 h HO/dioxane O hh 100°C, 2020 O0 0 -O O 100°C, o o o o o o

Boc, HN 0 Boc O IACDBoc N N) 0 0 T QO HN NH o 0 O BaOH N N N NH NaOH NaH ~ N~. OH DIACIN DIEA, CDI o NH TE'N TFA -0 N N NH N OH MeOHH 2 0,rt rt OH N LCHCN,7000 DCM,rt,rt,3 DCM 3 h 0 MeOH/HO, CHCN, 70°C 200 0 HCN 700 o 0 o 0 0 o o o o

Exemplary

[0521] Exemplary

[0521] Synthetic Synthetic Scheme Scheme for Exemplary for Exemplary Compound Compound 32: 32:

238

Boc BON Br o 0 o BocN N 0 Br Br Br- H2SO4 RuPhosPd, KMn4, NaOH KMnO, NaOH Br OH OH H 2 SO4 Br A O/ o RuPhosPd, CsCsCO CO 2 3 N N O OH 0 o CN H 2 0/dioxane HO/dioxane OH MeOH, 70 C, MeOH, 70°C, 2020 h h O PhMe, 16 hh reflux, 16 PhMe,reflux, Os o 0 0 o O 100°C, 20 100°C, 20 hh / 0 O o 0 O AO o O

Boc'N TFA Boc 0Boc'N Boc O N o N HN HN O NaOH N OH DIEA, CDI CDI N o o o NN NH NaOH N DIEA, ANH TFA OH N NH NH 2023248067 0 N o N MeOH/H 2 0, MeOH/HO, rtrt I OH CH 3CN,70°C CHCN, 70 C ,rt, 33 hh DCM,rt, DCM N O .1 o 0 o 0 o 110o o0 o

O O O

O H H KNN N No N N 0 0 o NHH NC NC T N N'-NN O NC ON O N A N H H N 0 N o o 0 NC NC A. Ny WINN N N CI CI O'

CI0 o o 100o CI NaBH(OAc), NaBH(OAc), DCM,DCM, rt, rt, 1h 1h O

1. Synthesisofof5-bromo-3-methoxybenzene-1,2-dicarboxylic

[0522] 1. Synthesis

[0522] acid 5-bromo-3-methoxybenzene-1,2-dicarboxylic acid

[0523] Into

[0523] Into a 100-mL a 100-mL round-bottom round-bottom flask, was flask, was placed placed 4-bromo-2-methoxy-6 4-bromo-2-methoxy-6-

methylbenzonitrile (800 mg, methylbenzonitrile (800 mg,3.54 3.54mmol, mmol, 1.00 1.00 equiv),water equiv), water (10mL), (10 mL), sodium sodium hydroxide hydroxide (708 (708 mg, mg,

17.70 mmol,5.00 17.70 mmol, 5.00equiv), equiv), KMnO KMnO4 (1.12 (1.12 g, 7.09 g, 7.09 mmol, mmol, 2.00 2.00 equiv). equiv). The resulting The resulting solution solution was was

stirred for 16 stirred for 16 hhatat 100°C 100°Cin in an an oiloil bath. bath. TheThe solids solids were were filtered filtered out.pHThe out. The pHofvalue value of the solution the solution

was adjusted was adjusted to to 33 with with hydrogen chloride(2(2 mol/L). hydrogen chloride mol/L). The Theresulting resulting solution solution was wasextracted extracted with with dichloromethane (15mLmL dichloromethane (15 x 3) X 3) andand thethe aqueous aqueous layers layers combined. combined. The resulting The resulting solution solution was was

extracted extracted with with ethyl ethyl acetate acetate/ /methanol methanol == 10:1 10:1 (15 (15 mL 3) and mL xx 3) and the the organic organic layers layers combined and combined and

dried in in an an oven oven under reduced pressure, under reduced pressure, concentrated concentrated under undervacuum. vacuum.This This resultedinin330 resulted 330mgmg (34%) of5-bromo-3-methoxybenzene-1,2-dicarboxylic (34%) of 5-bromo-3-methoxybenzene-1,2-dicarboxylic acida as acid as a white white solid. solid.

[0524] 2. 2.

[0524] Synthesisofof1,2-dimethyl Synthesis 1,2-dimethyl5-bromo-3-methoxybenzene-1,2-dicarboxylate 5-bromo-3-methoxybenzene-1,2-dicarboxylate

[0525] Into

[0525] Into a 100-mL a 100-mL round-bottom round-bottom flask, was flask, was placed placed 5-bromo-3-methoxybenzene-1,2 5-bromo-3-methoxybenzene-1,2-

dicarboxylic acid dicarboxylic acid (330 mg, mg, 1.20 1.20 mmol, mmol,1.00 1.00equiv), equiv),methanol methanol(20(20 mL), mL), sulfuric sulfuric acid(5(5mL). acid mL).TheThe resulting solution resulting solutionwaswas stirred stirred forfor 16ath 70°C 16 h at 70°C in aninoil anbath. oil bath. The resulting The resulting solution solution was was diluted diluted with water with water (40 (40 mL). mL). The ThepHpH valueofof value thesolution the solutionwas wasadjusted adjustedtoto8 8with withsodium sodium carbonate.TheThe carbonate.

resulting solution resulting solution was was extracted extracted with with ethyl ethyl acetate acetate(30 (30mL x 3) mL x 3) and and the the organic organic layers layers combined combined

and dried over and dried anhydroussodium over anhydrous sodium sulfateand sulfate andconcentrated concentratedunder under vacuum. vacuum. The The residue residue was was

239 applied ontoa asilica silicagelgelcolumn column with with ethyl ethyl acetate/petroleum etherThis (1:10). Thisinresulted in 340 2023248067 10 Oct 2023 applied onto acetate/petroleum ether (1:10). resulted 340 mg(93%) mg (93%)ofof1,2-dimethyl 1,2-dimethyl5-bromo-3-methoxybenzene-1,2-dicarboxylate 5-bromo-3-methoxybenzene-1,2-dicarboxylate as a white as a white solid.solid.

[0526] LC-MS

[0526] LC-MS (ES+): (ES+): m/z m/z 302.85 302.85 [MH+],

[MH+], = 0.906 tR =tR0.906 minmin (2.0minute (2.0 minuterun). run).

[0527] 3. Synthesis

[0527] 3. Synthesis of 1,2-dimethyl-5-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]-3 of 1,2-dimethyl-5-[4-[(tert-butoxy)carbonyl|piperazin-1-yll-3-

methoxybenzene-1,2-dicarboxylate methoxybenzene-1,2-dicarboxylate

[0528] Into Into

[0528] a 100-mL a 100-mL round-bottom flask, flask, round-bottom was placed was placed 1,2-dimethyl 1,2-dimethyl 5-bromo-3 5-bromo-3-

methoxybenzene-1,2-dicarboxylate methoxybenzene-1,2-dicarboxylate (300 (300 mg, mg, 0.990.99 mmol, mmol, 1.00 1.00 equiv), equiv), tert-butyl tert-butyl piperazine-1 piperazine-1-

carboxylate (277 (277 mg, mg,1.49 1.49mmol, mmol, 1.50 1.50 equiv),RuphosPd equiv), RuphosPd (39 (39 mg, mg, 0.050.05 mmol, mmol, 0.05 equiv), 0.05 equiv),

(978(978 Cs2CO3 CsCO 3.00 3.00 mg, mg, mmol, mmol, 3.00 equiv), 3.00 equiv), toluene toluene (15 mL). (15 mL). The resulting The resulting solution solution was stirred was stirred for for 12 12 hh at at 100°C 100°C inin anan oiloil bath.TheThe bath. resulting resulting solution solution was diluted was diluted with(30 with water water mL). (30 mL). The resulting The resulting

solution solution was extracted with was extracted ethyl acetate with ethyl acetate (30 (30 mL 3) and mL xx 3) the organic and the organic layers layers combined anddried combined and dried over anhydroussodium over anhydrous sodium sulfateand sulfate andconcentrated concentratedunder under vacuum. vacuum. The The residue residue was was applied applied onto onto a a silica silicagel gelcolumn column with with dichloromethane/ethyl acetate (10:1). dichloromethane/ethyl acetate (10:1). This resulted resulted in in 340 340 mg (84%)ofof mg (84%)

1,2-dimethyl 5-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylate 5-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylate as light yellow as light yellowoil. oil.

[0529] LC-MS

[0529] LC-MS (ES+): (ES+): m/z m/z 409.05 409.05 [MH+],

[MH+], = 0.963 tR =tR0.963 minmin (2.0minute (2.0 minuterun). run).

[0530] 4. 4.

[0530] Synthesis 5-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene Synthesisofof5-[4-[(tert-butoxy)carbonyl|piperazin-1-yl]-3-methoxybenzene- 1,2-dicarboxylic acid 1,2-dicarboxylic acid

[0531] Into Into

[0531] a 100-mL a 100-mL round-bottom flask, flask, round-bottom was placed was placed 1,2-dimethyl 1,2-dimethyl 5-[4-[(tert 5-[4-[(tert-

butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylate butoxy)carbonyl|piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylate (340(340 mg, 0.83 mg, 0.83 mmol,mmol, 1.00 1.00

equiv), methanol/H20/THF equiv), (8 mL), methanol/HO/THF (8 mL), sodiumol sodiumol (1002.50 (100 mg, mg,mmol, 2.50 3.00 mmol, 3.00 equiv). equiv). The resulting The resulting

solution was stirred for was stirred for12 12hhatat25°C 25°CThe Theresulting resultingsolution solutionwas wasdiluted dilutedwith withwater water (30 (30 mL). mL). The The

pHvalue pH valueof of thethe solution solution was was adjusted adjusted to 8hydrogen to 8 with with hydrogen chloride chloride (2 mol/L). (2 mol/L). citric acid citric acid monohydratewas monohydrate was employed employed to adjust to adjust thethe pH pH to 3. to 3. TheThe resulting resulting solution solution was was extracted extracted with with ethyl ethyl

acetate acetate (30 (30 mL 3) and mL xx 3) and the the organic organic layers layers combined anddried combined and driedover overanhydrous anhydrous sodium sodium sulfate sulfate

and concentrated and concentrated under undervacuum. vacuum.This This resultedinin300 resulted 300mgmg (95%) (95%) of 5-[4-[(tert of 5-[4-[(tert-

butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylic butoxy)carbonyl]piperazin-1-yl]-3-methoxybenzene-1,2-dicarboxylic acid acid as colorless as colorless oil. oil.

[0532] LC-MS

[0532] LC-MS (ES+): (ES+): m/z m/z 306.95 306.95 [MH+],

[MH+], = 0.853 tR =tR0.853 minmin (2.0minute (2.0 minuterun). run).

[0533] 5. Synthesis

[0533] 5. Synthesis of tert-butyl-4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3 of tert-butyl-4-|2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3-

dihydro-1H-isoindol-5-yl]piperazine-1-carboxylate dihydro-1H-isoindol-5-yllpiperazine-1-carboxylate

240

100-mL 2023248067 10 Oct 2023

[0534] Into Into

[0534] a a 100-mL round-bottom round-bottom flask, flask, was placed was placed tert-butyl tert-butyl 4-(7-methoxy-1,3-dioxo-1,3 4-(7-methoxy-1,3-dioxo-1,3-

(260 dihydro-2-benzofuran-5-yl)piperazine-1-carboxylate(260 dihydro-2-benzofuran-5-yl)piperazine-1-carboxylate mg,mg, 0.72 0.72 mmol, mmol, 1.00 1.00 equiv), equiv), 3- 3 aminopiperidine-2,6-dionehydrochloride aminopiperidine-2,6-dione hydrochloride(153.6 (153.6mg,mg, 0.93 0.93 mmol, mmol, 1.301.30 equiv), equiv), pyridine pyridine (10 (10 mL).mL).

Theresulting The resultingsolution solution waswas stirred stirred forh 4at h120°C for 4 at 120°C in an in oilan oil bath. bath. The resulting The resulting solutionsolution was was diluted diluted with with water water (30 (30 mL). Theresulting mL). The resulting solution solution was extracted with was extracted with ethyl ethyl acetate acetate (30 (30 mL 3) mL Xx 3)

and the organic layers and the layers combined anddried combined and driedover overanhydrous anhydrous sodium sodium sulfate sulfate andand concentrated concentrated

under vacuum. under vacuum.The The residuewaswas residue applied applied onto onto a silicagel a silica gelcolumn columnwith with dichloromethane/methanol dichloromethane/methanol

(100:1). (100:1). This This resulted resulted in in280 280 mg (83%)ofoftert-butyl mg (83%) tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy 4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy-

1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1-carboxylateas as 1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1-carboxylate a yellow a yellow solid. solid.

[0535] LC-MS

[0535] LC-MS (ES+): (ES+): m/z m/z 417.05 417.05 [MH+],

[MH+], = 0.852 tR =tR0.852 minmin (2.0minute (2.0 minuterun). run). 6. Synthesis

[0536] 6. Synthesis

[0536] of 2-(2,6-dioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1 of 2-(2,6-dioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1-

yl)isoindoline-1,3-dione yl)isoindoline-1,3-dione

[0537] Into Into

[0537] a 50-mL a 50-mL round-bottom flask, flask, round-bottom was placed was placed tert-butyl tert-butyl 4-[2-(2,6-dioxopiperidin-3 4-[2-(2,6-dioxopiperidin-3-

yl)-7-methoxy-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1-carboxylate yl)-7-methoxy-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1-carboxylate (270(270 mg, mg, 0.570.57

mmol,1 1equiv), mmol, equiv), dichloromethane dichloromethane(6 (6mL, mL, 0.07 0.07 mmol, mmol, 0.124 0.124 equiv), equiv), TFA TFA (2 0.02 (2 mL, mL, mmol, 0.02 mmol, 0.031 equiv).TheThe 0.031 equiv). resulting resulting solution solution was stirred was stirred for 2 for 2 hr hr at at 25 25 °C. The°C. The resulting resulting mixture was mixture was

concentrated to give concentrated to give 2-(2,6-dioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1-yl)isoindoline-1,3- 2-(2,6-dioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1-yl)isoindoline-1,3 dione asa abrown dione as brownoil.oil.

[0538] LC-MS

[0538] LC-MS (ES+): (ES+): m/z m/z 373.05 373.05 [MH+],

[MH+], = 0.155 tR =tR0.155 minmin (2.0minute (2.0 minuterun). run).

[0539] 7. Synthesis

[0539] 7. Synthesis of 6-[4-([4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3 of 6-|4-([4-|2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3-

dihydro-1H-isoindol-5-yl]piperazin-1-yl]methyl)piperidin-1-yl]-N-[(1r,4r)-4-(3-chloro-4 dihydro-1H-isoindol-5-yl|piperazin-1-yl|methyl)piperidin-1-yl-N-|(1r,4r)-4-(3-chloro-4-

cyanophenoxy)cyclohexyl]pyridazine-3-carboxamide cyanophenoxy)cyclohexyl|pyridazine-3-carboxamide

[0540] Into Into

[0540] a 100-mL a 100-mL round-bottom round-bottom flask, flask, was placed was placed 2,2,2-trifluoroacetaldehyde; 2,2,2-trifluoroacetaldehyde; 2-(2,6 2-(2,6-

dioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione (130 lioxopiperidin-3-yl)-4-methoxy-6-(piperazin-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione(130. mg,mg,

0.28 mmol, 0.28 mmol,1.078 1.078equiv), equiv),dichloromethane dichloromethane(10(10 mL,mL, 0.12 0.12 mmol), mmol), 6-(4-formylpiperidin-1-yl)-N 6-(4-formylpiperidin-1-yl)-N-

[(1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl]pyridazine-3-carboxamide

[(1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl]pyridazine-3-carboxamide (120 (120 mg, mmol, mg, 0.26 0.26 mmol, 1 equiv), equiv), NaBH(OAc)3 (163.4 NaBH(OAc) (163.4 mg, mg, 0.77 0.77 mmol, mmol, 3.006 3.006 equiv). equiv). The resulting The resulting solution solution was stirred was stirred

for for 22 hr hr at at 25°C. 25°C. The resulting solution The resulting solution was was diluted diluted with with dichloromethane (30mL). dichloromethane (30 mL).The The resulting mixture resulting mixture was washedwith was washed withHOH(30 2 0 (30 mL xmL 3). The 3).x The mixture mixture was dried was dried over anhydrous over anhydrous

sodium sulfate and sodium sulfate and concentrated concentrated under undervacuum. vacuum.TheThe resulting resulting mixture mixture waswas concentrated concentrated under under

vacuum. Theresidue vacuum. The residuewas wasapplied appliedonto ontoa silica a silica gel gel column columnwith withdichloromethane/ethyl dichloromethane/ethyl acetate acetate

241

(3:1). (3:1). The The crude crude product waspurified product was purifiedbybyPrep-HPLC Prep-HPLCwithwith the the following following conditions: conditions: Column, Column,

XBridge Prep C18 XBridge Prep OBDColumn, C18 OBD Column,5um, 5um,19*150mm; mobile 19*150mm; mobile phase,Water phase, Water(10 (10 mmol/L mmol/LNH4HCO) NH 4HCO 3

) and acetonitrile (43% and acetonitrile PhaseB Bupup (43% Phase to to 65% 65% in min); in 8 8 min); Detector, Detector, uv.uv. This This resultedinin7070mgmg resulted

(33.11%) of6-[4-([4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3-dihydro-1H- (33.11%) of 6-[4-([4-[2-(2,6-dioxopiperidin-3-yl)-7-methoxy-1,3-dioxo-2,3-dihydro-1H isoindol-5-yl]piperazin-1-yl]methyl)piperidin-1-yl]-N-[(1r,4r)-4-(3-chloro-4 isoindol-5-yl]piperazin-1-yl]methyl)piperidin-1-yl]-N-|(1r,4r)-4-(3-chloro-4-

cyanophenoxy)cyclohexyl]pyridazine-3-carboxamide as a yellow cyanophenoxy)cyclohexyl]pyridazine-3-carboxamide as a yellow solid. solid. 2023248067

[0541] 1H NMR

[0541] MHz, MHz, (400 (400 1H NMR DMSO-d6) DMSO-d6) 6 11.04 11.04 (s, 1H), 8.57 J(d,= J= 1H), (d, (s, 8.57 8.4Hz, 8.4Hz, 1H), 1H), 7.87-7.79 7.87-7.79

(m, 2H),7.39-7.32 (m, 2H), 7.39-7.32(m, (m,2H), 2H), 7.15-7.12 7.15-7.12 (m,(m, 1H), 1H), 6.966.96 (s, 1H), (s, 1H), 6.686.68 (s, (s, 1H), 1H), 5.04-4.98 5.04-4.98 (m, (m, 1H), 1H),

4.50-4.47 (m, 4.50-4.47 (m, 3H), 3H), 4.93-3.85 4.93-3.85(m, (m,4H), 4H),3.35-3.33 3.35-3.33(m,(m, 5H), 5H), 3.07 3.07 - 2.81 - 2.81 (m,(m, 3H), 3H), 2.51 2.51 (s,(s,3H), 3H), 2.27 -22.1 2.27 -22.1 (m, (m, 2H), 2H), 2.09 2.09-2.01 -2.01 (m, (m,2H), 2H),2.00 2.00- -1.49 1.49(m, 11H),1.23 (m,11H), 1.23- -1.11(m, 1.11(m,3H); 3H); LC-MS LC-MS

(ES+): m/z m/z 824.25/826.25 824.25/826.25[MH+],

[MH+], tR = 182 tR 182 = minmin (3.0(3.0 minute minute run). run).

[0542] Chemical

[0542] ChemicalFormula: Formula: C 4 2 H 4 6ClN CHCINO 90 7 [823.32/825.32]

[823.32/825.32]

[0543] Total

[0543] Total H count HNMR fromHNMR H countfrom data: 46.46. data:

[0544] Exemplary

[0544] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 34 Compound 34

CI CI O 0 00 o o o HN, NH N CO N NN N O N 0 N N N _, N rac-N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-((4-(2'-(2,6 rac-N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(4-(2-(26-

dioxopiperidin-3-yl)-3'-oxospiro[cyclopropane-1,1'-isoindolin]-6'-yl)piperazin-1 dioxopiperidin-3-yl)-3'-oxospirolcyclopropane-1,1'-isoindolin|-6'-y)piperazin-1-

yl)methyl)piperidin-1-yl)benzamide yl)methyl)piperidin-1-yl)benzamide

[0545] Synthetic

[0545] scheme: Synthetic scheme:

242

Oct 2023 00 1 0012 ,70°C 1.SOCl,70 C 0 0

HO H0 0 HO HO OH 2.Br,70°C OH 2.Br 2 ,70C B 3.MeOH,0°C 3.MeOH,0 °C Br Br

0 O 0 o 0 0/ 0 -NH NH o0 O 0 0N, o o Ii I Br NN 0( Br .NBo Boc NTi(i-PrO) NH Ti(i-PrO)4 4 N NH N DIEA, DMSO DIEA, DMSO N N THF, EtMgBr THF, EtMgBr NN NaH,DMF, NaH, DMF,3030°C, °C1212h h BcN Boc N F N 2023248067 10 NNN eCN N 90 °Ca3Th12h N Boc

M~aN H o 0 0 ocBcoc 0 0 0 0 N, OH OH CNH N OH OH 0 NaOH,THF N UreaNMPN NH2 C) N , N NH NaOH,THF N Urea, NMP N N NaBHAO), T DCE,+ + N 12.50h N N N 3N + N N + MeOH, H2O H 20 3N.0 160 °C, 2h CN_ N MeOH, Bo N OH Boc' N Br OH Boc' N NH r BC N Boc Boc Boc OH OH NH2 Boc Bo- NH0N o o' o

0 0 OH o OH urH O 0 m o OH O o00 o 1b N N (629 g, 1 NH Hes ldioxane HCl/dioxane NH NH N' N 0 C)I N-L N 0= N N,,MeCN, 90 *C.3h MeCN, 90 °C, 3h N'>r N N Bie N w t NH Boc HN.> BToeCN N NH2 HN Boc O

O NH N CI N CI 0 o N HN N NH NH NH, Nish N m 0,73mL 8 NBH(AcO), NaBH(AcO), TEA,TEA, CE30 DCE, 30 C, 12 °C, 12.5 h N °CN. Na Lt N desNie 0 N N O CI N

105461

[0546] Step 1: Step 1: Synthesisof Synthesis of dimethyl 2-bromopentanedioate dimethyl 2-bromopentanedioate

0 o 0 o 11.SOCI,70 SOCI C 2 ,70 °C 0 o 0 o

HOH2.Br 2 ,7000C 0 HO OH 2.Br,70 °C 3.MeOH,O0'C0 3.MeOH,0 °C Br1 Br

[0547] To a Toasolution

[0547] ofglutaricacid(30g, solution of glutaric 227.07mmol, acid (30 g, 227.07 mmol, 1 eq) Iineq)inchloroform(90mL)was chloroform (90 mL) was

addedthionylchloride(59g,499.56mmol,36mL,2.2eq).Themixturewasstirredat70'Cfor added thionyl chloride (59 g, 499.56 mmol, 36 mL, 2.2 eq). The mixture was stirred at 70 °C for

1 1 h. h.Liquid Liquid bromine (36.29 g, bromine (36.29 g, 227.07 mmol,1Ieq) 227.07 mmol, wasadded 1 eq) was addedinto intothe themixture mixturedropwise. dropwise.TheThe mixturewas mixture wasstirred stirred at at 70'C 70 °C for for 12 12h.The h. Themixture mixture was cooled to was cooled to 0'C 0 °C and and methanol (58g,g, 1.82 methanol (58 1.82 mol, 73 mol, 73 mL, mL,88eq) wasadded eq) was addedinto intothe the mixture mixturedrop dropwise wiseatat0 'C.LCMS 0 °C. detected LCMS detected thethe desired desired

product. The product. mixturewas The mixture wasextracted extractedwith withethyl ethylacetate acetate(150 (15 0mL mL X x3) andwashed 3) and washed with with saturated saturated

aqueous sodium aqueous sodium bicarbonate bicarbonate (200 (200 mL). mL). TheThe organic organic layer layer waswas dried dried over over anhydrous anhydrous sodium sodium

sulfate sulfate and and concentrated. concentrated. The The residue residue was was purified purified with with Flash Flash C18 C18 column chromatography column chromatography

(acetonitrile: (acetonitrile:water water= =1:0 1:0toto 1:1). Dimethyl 1:1). 2-bromopentanedioate Dimethyl (4g+20g g(crude), 2-bromopentanedioate (4g+20 16.73 mmol, (crude), 16.73 mmol, 700yield) wasobtained 7% yield) was obtainedasasaayellow oil. yellow oil.

105481 LCMS:

[0548] LCMS: MS MS (ESI) (ESI) m/z:241.0 m/z: 241.0 [M+1>'.

[M+1]. Chemical 105491 Chemical

[0549] Formula: Formula: lBrO4, Molecular C 7 1 Molecular CHBrO, Weight:Weight: 239.0623 9.06

243

2023 [0550]

[0550] ¹H 1H NMR: NMR: (400 (400 DCCl)DCCl3) MHz, MHz, .: 4.36 : 4.39 - - 4.36 4.39 (m, 1H), 1H),(s, (m,3.78 3.78 (s, 3.72 3H), 3H), (s, 3H), -2.56 (s, 2.56 3.723H), 2.49 (m, 2.49 (m,2H), 2H),2.44 2.44 - 2.34 - 2.34 (m, (m, 1H), - 2.23- (m, 1H), 2.33 2.33 2.231H). (m, 1H). 2023248067 10 Oct

[0551]

[0551] Total H Total H count HNMR fromHNMR count from data: 11. 11. data:

[0552] StepStep

[0552] 2: 2: Synthesisofoftert-butyl Synthesis tert-butyl 4-(3-cyano-4-(methoxycarbonyl) 4-(3-cyano-4-(methoxycarbonyl)

phenyl)piperazine-1-carboxylate phenyl)piperazine-1-carboxylate

0 o NH O N / Boc' Boc O

DIEA, DMSO N F F DIEA, DMSO N N N N N Boc' N N Boc

[0553] To a To

[0553] a solution solution of methyl of methyl 2-cyano-4-fluoro-benzoate (10 g,(10 2-cyano-4-fluoro-benzoate g, 55.82 55.82 1 eq), 1tert- mmol, mmol, eq), tert butyl piperazine-1-carboxylate butyl (12.48 g, piperazine-1-carboxylate (12.48 g, 66.98 66.98 mmol, mmol,1.2 1.2eq) eq)inin dimethylsulfoxide dimethylsulfoxide(100 (100mL) mL) waswas

added diisopropylethylamine added disopropylethylamine (28.86 (28.86 g, g, 223.28 223.28 mmol, mmol, 4 eq). 4 eq). The The reaction reaction mixture mixture was was stirred stirred at at

120 °C for 120 °C for 12 12 h. h. Thin layer chromatography Thin layer (petroleum chromatography (petroleum ether:Ethyl ether: Ethylacetate acetate==3:1) 3:1) showed showed methyl 2-cyano-4-fluoro-benzoate methyl 2-cyano-4-fluoro-benzoatewaswas consumed, consumed, and desired and desired product product was detected. was detected. The The mixture was mixture waspoured pouredinto intowater water(50 (50mL), mL),andand filtered. The filtered. Thefiltrate filtrate was was dried dried under under vacuum. The vacuum. The

residue was residue purified with was purified silica gel with silica gelcolumn column chromatography (petroleum chromatography (petroleum ether:ethyl ether: ethylacetate acetate == 10:1 10:1 to to 3:1). 3:1).Tert-butyl Tert-butyl4-(3-cyano-4-methoxycarbonyl-phenyl)piperazine-1- carboxylate 4-(3-cyano-4-methoxycarbonyl-phenyl)piperazine-1- carboxylate (18(18 g, g,

52.11 mmol,93% 52.11 mmol, 93% yield)waswas yield) obtained obtained as as a yellow a yellow solid. solid.

[0554] Chemical

[0554] Formula: C ChemicalFormula: 1 8 H23N CHNO, 3 0 4 , Molecular Molecular Weight: Weight: 345.39 345.39

[0555] Step Step

[0555] 3: Synthesis 3: Synthesis of tert-butyl of tert-butyl 4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5' 4-(1'-oxospiro|cyclopropane-1,3'-isoindoline|-5'-

yl)piperazine-1-carboxylate yl)piperazine-1-carboxylate

0 o 0 O O Ti(i-PrO)4 Ti(i-PrO) NH NH N N -N THF, EtMgBr THF, EtMgBr N N N N BocNON o N Boc

[0556] To a To

[0556] a solution solution of tert-butyl of tert-butyl 4-(3-cyano-4-methoxycarbonyl-phenyl)piperazine-1 4-(3-cyano-4-methoxycarbonyl-phenyl)piperazine-1-

carboxylate (18 g, carboxylate (18 g, 52.11 52.11 mmol, mmol, 1 1eq) eq)inin tetrahydrofuran tetrahydrofuran (200 (200mL) mL)was was added added tetraisopropyl tetraisopropyl

titanate titanate (17.77 (17.77 g,g,62.54 62.54mmol, 1.2 eq) and mmol, 1.2 and aa solution solution of ofethyl ethylmagnesium bromideinin magnesium bromide

tetrahydrofuran (2 (2 M, 52.11 mL, M, 52.11 mL,2 2eq) eq)atat 00 °C. °C. The Themixture mixturewas wasstirred stirredat at 25 25 °C °C for for 1 h. h. Thin Thin

layer chromatography (petroleum chromatography (petroleum ether:ethyl ether: ethylacetate acetate ==1:1) 1:1) showed showedtert-butyl tert-butyl 4-(3-cyano-4- 4-(3-cyano-4 methoxycarbony 1-phenyl)piperazine-1-carboxylate methoxycarbony 1-phenyl)piperazine-1-carboxylate was was consumed, consumed, and desired and desired product product was was detected. The detected. mixture was The mixture wasadded addedinto intosaturated saturated aqueous aqueousammonuim ammonuim chloride chloride (150 (150 mL). mL). The The

244 mixture was wasextracted withethyl extracted with ethyl acetate acetate (100 (100 mL mLx x3). 3). The Theorganic layerwas organiclayer wasdried driedover oversodium sodium 2023248067 10 Oct 2023 mixture sulfate andconcentrated. sulfate and concentrated.The The residue residue was triturated was triturated withacetate with ethyl ethyl acetate (30filtered. (30 mL) and filtered. Tert mL) andTert- butyl 4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-l-carboxylate butyl 4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-1-carboxylate(6 g, 17.47 (6 g, 17.47 mmol, 33% mmol,33% yield)was yield) was obtained obtained as as a yellow a yellow solid. solid.

[0557] Chemical

[0557] Formula: C ChemicalFormula: 19 H 2 5 0 CHON, 3N 3 , Molecular Molecular Weight: Weight: 343.42 343.42

[0558] ¹H NMR:

[0558] CDCl)CDCl MHz, MHz, (400 (400 1H NMR: 3) : 7.75 7.73- (d, -: 7.75 1H),1H), Hz, Hz, (d, J=8.8 7.73J=8.8 6.976.97 - 6.95(d, - 6.95 (d, J=8.8 Hz, 1H), Hz, 1H), 6.94 6.94 - 6.85 - 6.85 (m, (m, 1H), 1H), 6.41 6.41 (s, 3.61 3.58 (s, 1H), 1H), - 3.58 3.61(t, J=4.8(t, Hz,J=4.8 Hz, -4H), 4H), 3.28 3.253.28 - 3.25Hz,(t, (t, J=4.8 J=4.8 Hz, 4H), 1.56 4H), 1.56(s,(s,2H), 2H),1.49 1.49 (s,(s, 9H), 9H), 1.381.38 - 1.36 - 1.36 (m, (m, 2H). 2H).

[0559] Total

[0559] Total H count HNMR fromHNMR H countfrom data: 25.25. data:

[0560] Step Step

[0560] 4: Synthesis 4: Synthesis of dimethyl of dimethyl 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedioate spiro[cyclopropane-1,1'-isoindoline]-2'-yl|pentanedioate

0 0 0 0 0 o 0 0 o -O NH Br N 0 N NN H NaH, DMF, NaH, DMF,3030°C, °C,1212h h N o N Boc N o 0 0 o O

[0561]

[0561] 20 parallel: batchesininparallel: 20 batches

[0562]

[0562] To To aasolution solutionofof tert-butyl tert-butyl4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5' 4-('-oxospiro[cyclopropane-1,3'-isoindoline]-5' yl)piperazine-1- carboxylate yl)piperazine-1. carboxylate (100 (100 mg, mg, 0.29 0.29mmol, mmol,1 1eq)eq)and anddimethyl dimethyl 2-bromopentanedioate 2-bromopentanedioate

(104 mg, 0.44 (104 mg, 0.44 mmol, mmol,1.51.5eq) eq)inindimethylformamide dimethylformamide (2 mL) (2 mL) was was addedadded sodiumsodium hydride hydride (35 mg,(35 mg,

0.88 mmol, 60% mmol, 60% in in mineral mineral oil,3 3eq). oil, eq). The Themixture mixturewas wasstirred stirredatat 30 30 °C °C for for 12 12 h. h. Thin layer Thin layer

cromatography (petroleum cromatography (petroleum ether:ethyl ether: ethylacetate acetate ==1:1) 1:1) showed showed30% 30% of the of the tert-butyl4-(1'- tert-butyl 4-(1' oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-1-carboxylate was oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-1-carboxylate wasconsumed. consumed. TheThe 20 20 reaction mixtures reaction were poured mixtures were pouredinto into 50 50 mL mLofofbrine, brine,and andextracted extractedwith withethyl ethyl acetate acetate (30 (30 mL mLx x2), 2), the combined the organiclayers combined organic layerswere weredried driedover overanhydrous anhydrous sodium sodium sulfate, sulfate, filteredand filtered andconcentrated concentrated in in vacuum. Theresidue vacuum. The residuewas waspurified purifiedbybysilica silica gel gel column chromatography column chromatography (petroleum (petroleum ether/ ether/

ethyl acetate=3/1 ethyl acetate=3/1to to1/1). 1/1).Dimethyl Dimethyl 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo 1 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedioate (200 mg, spiro[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedioate (200 mg, 0.40 0.40 mmol, mmol,10%10% yield yield

corrected forrecovered corrected for recovered starting starting material) material) was obtained was obtained as a oil. as a yellow yellow Also oil. Also was isolated isolated tert- was tert

butyl 4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-1-carboxylatet butyl 4-(1'-oxospiro[cyclopropane-1,3'-isoindoline]-5'-yl)piperazine-1-carboxylate (675 mg). (675 mg).

[0563] Formula: C ChemicalFormula:

[0563] Chemical C2HNO, 0 7 , Molecular 2 6H 3 5 N 3Molecular Weight: 501.57 Weight:501.57

245 of 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo 10 Oct 2023

[0564] Step Step

[0564] 5: Synthesis 5: Synthesis of 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedioicacid spiro[cyclopropane-1,1'-isoindoline]-2'-yl|pentanedioicacid

0 0 / o 0o 0 o OH OH N N NaOH,THF NaOH, THF N N N N0 N N/ MeoH, HHO MeOH, 20 N N Boc' N Boc O Boc' N Boc OH OH o 0 0 o O

[0565] To a To a solution of dimethyl 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro 2023248067

[0565] solution of dimethyl 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro

[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedioate (800

[cyclopropane-1,1'-isoindoline]-2'-yl]pentanedicate (800 mg, mg, 1.59 1.59mmol, mmol,1 eq) 1 eq)inin tetrahydrofuran (5 mL) tetrahydrofuran (5 andmethanol mL) and methanol(5 (5mL) mL) waswas added added a solution a solution of sodium of sodium hydroxide hydroxide (255 mg, (255 mg,

6.38 6.38 mmol, mmol, 44eq) eq)inin water water (3 (3 mL). mL). The Themixture mixturewas was 25 °C stirredatat25 stirred °Cfor for 22 hr. hr. LCMS showed LCMS showed the the

reaction was reaction completedand was completed anddesired desiredMSMS waswas detected. detected. TheThe mixture mixture together together withwith the the other other batch batch

was pouredinto was poured into 20 20mL mLwater, water,and andadjusted adjustedthe thepHpH to to 3.0with 3.0 with2.0 2.0N Nhydrochloride hydrochloride acid,then acid, then extracted extracted with with ethyl ethyl acetate acetate (30 (30 mL mL x 3). 3). The The combined organiclayers combined organic layerswere weredried driedover over anhydrous sodium anhydrous sodium sulfate,then sulfate, thenconcentrated concentratedininvacuum. vacuum.2-[6'-(4-tert-butoxycarbonylpiperazin-1- 2-[6'-(4-tert-butoxycarbonylpiperazin-1 yl)-3'-oxo-spiro[cyclopropane-1,'-isoindoline]-2'-yl]pentanedioic yl)-3'-oxo-spiro[cyclopropane-1,1'-isoindoline]-2'-yI]pentanedioic acid acid (740 (740 mg, 1.56 mmol, mg, 1.56 mmol, 97% yield) as 97% yield) as an an off-white off-white solid solid was obtained, which was obtained, whichwas wasdirectly directly used usedfor for the the next next step step without without

further purification. further purification.

[0566] LCMS:

[0566] LCMS: MS MS (ESI) (ESI) m/z:474.3[M+1]*. m/z: 474.3[M+1]'.

[0567] Chemical

[0567] Formula: C ChemicalFormula: 2 4 H 3 1N CHNO, 3 0 7 , Molecular Molecular Weight: Weight: 473.52 473.52

[0568] Step Step

[0568] 6: Synthesis 6: Synthesis of 5-amino-4-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo of 5-amino-4-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo-pentanoic acid; spiro[cyclopropane-1,1'-isoindoline|-2'-yl]-5-oxo-pentanoic acid; 5-amino-2-[6'-(4-tert 5-amino-2-[6'-(4-tert-

butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo butoxycarbonylpiperazin-1-yl)-3'-oxo-spirolcyclopropane-1,1'-isoindoline|-2'-yll-5-oxo-

pentanoic acidand pentanoic acid and tert-butyl4-|2'-(2,6-dioxo-3-piperidyl)-1'-oxo-spiro|cyclopropane-1,3'- tert-butyl 4-[2'-(2,6-dioxo-3-piperidyl)-1'-oxo-spiro[cyclopropane-1,3' isoindoline]- 5 '-yl]piperazine-1-carboxylate isoindoline|-5'-yl|piperazine-1-carboxylate

246

Oct 2023 0 0 o 0 0 o 0 0 o OH OH OH NH NH 2 N N Urea, NMP N N N N N OH N N N 160 °C, 2h + N N Boc'N OH 0 160 0,2h BocN N N Boc OH Boc OH OH Boc' Boc NH 2 0 NH O O 0 0 O 2023248067 10

o NH N o + +N N N0 N Boc'N Boc

[0569] A mixture

[0569] Amixtureof2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro[cyclopropane of 2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro[cyclopropane-

1,1'-isoindoline]-2'-yl]pentanedioic acid 1,1'-isoindoline]-2'-yl]pentanedioic acid(400 (400mg, mg, 0.85 0.85 mmol, mmol, 11 eq) eq) and (253 mg, urea (253 and urea mg, 4.22 4.22 mmol,5 5eq) mmol, 1-methyl-2-pyrrolidinone(4(4mL) eq)inin 1-methyl-2-pyrrolidinone mL)waswas heated heated to to 160160 °C °C and and stirred stirred at at 160 160 °C °C forfor 2 2 hours. LCMS hours. showed LCMS showed two two peaks peaks with with desired desired MS signals. MS signals. The mixture The mixture together together withother with the the other batch was batch was filtered. filtered. The The filtrate filtratewas wasfurther purified further by by purified Semi-preparative Semi-preparativereverse phase reverse phaseHPLC HPLC

(column: BostonGreen (column: Boston Green ODS ODS 150*30 150*30 5 um;5 mobile um; mobile phase:phase:

[water[water (0.225% (0.225% formic formic acid)- acid)

acetonitrile]; acetonitrile];B%: B%: 35%-45%, 35%-45%, 1010 min).2 2isomeric min). isomericmono-amides mono-amides 5-amino-4-[6'-(4-tert 5-amino-4-[6'-(4-tert-

butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro[cyclopropane-1,1-isoindoline]-2'-yl]-5-oxo-

pentanoic acid pentanoic acid and and 5-amino-2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo- 5-amino-2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo spiro[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo-pentanoic acid were spiro[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo-pentanoic acid obtained(170 were obtained (170 mg, mg,0.36 0.36 mmol,42% mmol, 42% yieldandand yield 90 90 mg,mg, 0.19 0.19 mmol, mmol, 22% 22% yield yield respectively. respectively. It was It was not not conclusively conclusively

established which established which of the of the 2 isomeres 2 isomeres corresponds corresponds to whichto which structure.) structure.) Alsowasisolated Also isolated was tert-butyl tert-butyl

4-[2'-(2,6-dioxo-3-piperidyl)-l'-oxo-spiro[cyclopropane -1,3'-isoindoline]-5'-yl]piperazine-1 -|2-(2,6-dioxo-3-piperidyl)-1'-oxo-spiro[cyclopropane-1,3'-isoindoline]-5'-yllpiperazine-1-

carboxylate (90 mg, carboxylate (90 mg, 0.20 0.20mmol, mmol,23% 23% yield) yield) as as an an off-whitesolid. off-white solid. LCMS:

[0570] LCMS:

[0570] mono-amide mono-amide product m/z: m/z: MS (ESI) 1: MS1:(ESI) product 473.1[M+1] 473.1[M+1] Mono-amide productproduct *, Mono-amide 2: 2: MS (ESI) m/z: MS (ESI) m/z: 473.1[M+1]*, 473.1[M+1] Imide *, Imide product product 3: MS 3: MS (ESI) (ESI) m/z: m/z: 455.1[M+1] *. 455.1[M+1]*.

[0571] Chemical

[0571] Chemical Formula Formula mono-amide mono-amide product product 1: CHNO, MolecularMolecular 1: C24H32N406, Weight: Weight: 472.53. 472.53.

[0572] Chemical

[0572] Chemical Formula Formula mono-amide mono-amide product product 2: CHNO, MolecularMolecular 2: C24H32N406, Weight: Weight: 472.53. 472.53.

[0573] Chemical

[0573] Chemical Formula Formula Imide Imide product: C 2 4H Molecular product:CHNO, 454.52.454.52. Weight: Weight: 30 N 4 0 5 , Molecular

[0574] Step Step

[0574] 7a: Synthesis 7a: Synthesis of 3-(3'-oxo-6'-piperazin-1-yl-spiro[cyclopropane of 3-(3'-oxo-6'-piperazin-1-yl-spiro|cyclopropane-1,1'- -1,1' isoindoline]-2'-yl)piperidine-2,6-dione fromthethemono-amide isoindoline|-2'-yl)piperidine-2,6-dione from mono-amide product product 1 of 6step 6 1 of step

247

Oct 2023 0 O 0 % OH OH OH OH I_\b0 O o 0 o N NH N N H ON N 0 N MeCN, 90 °C, 3 h N MeCN, 90 °C, 3 h N N Boc' N Boc1 NH 2 HN 0 NH HN O 2023248067 10

[0575] To a To

[0575] a mixture mixture of 5-amino-2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spiro of 5-amino-2-[6'-(4-tert-butoxycarbonylpiperazin-1-yl)-3'-oxo-spird

[cyclopropane-1,1'-isoindoline]-2'-yl]-5-oxo-pentanoic acid

[cyclopropane-1,1'-isoindoline]-2-yl]-5-oxo-pentanoic acid(190 (190mg, mg,0.40 0.40 mmol, mmol, 1 eq, 1 eq, thethe first first

eluting eluting mono-amide product mono-amide product from from above) above) in acetonitrile(15 in acetonitrile (15mL) mL)waswas added added benzenesulfonic benzenesulfonic acid acid

(114 mg, 0.72 (114 mg, 0.72 mmol, mmol,1.80 1.80eq)eq)ininone oneportion portionatat 25 25 °C °Cunder undernitrogen nitrogenatmosphere. atmosphere.TheThe mixture mixture

was stirred was stirred at at90 90 °C °C for for3 3hours. hours.LCMS showed LCMS showed thethe product product waswas thethe main main peak. peak. TheThe mixture mixture was was concentrated in vacuum. concentrated in vacuum.The Theresidue residuewas waspurified purifiedbybySemi-preparative Semi-preparative reverse reverse phase phase HPLC HPLC

(column: BostonGreen (column: Boston Green ODS ODS 150*30 150*30 5 um;5 mobile um; mobile phase:phase:

[water[water (0.225% (0.225% formic formic acid)- acid)

acetonitrile]; B%: acetonitrile]; B%: 1%-27%, 1%-27%, 1010min). min).The The product product 3-(3'-oxo-6'-piperazin-1-yl 3-(3'-oxo-6'-piperazin-1-yl-

spiro[cyclopropane-1,1'-isoindoline]- 2'-yl)piperidine-2,6-dione spiro[cyclopropane-1,1'-isoindoline]- 2'-yl)piperidine-2,6-dione ( (55 mg,(55 mg, 0.14 0.14 mmol, 34%mmol, yield, 34% yield,

benzene sulfonate) benzene sulfonate) was wasobtained obtainedasasaabrown brownsolid. solid.

[0576] LCMS:

[0576] LCMS: EW4875-628-P1B, EW4875-628-P1B, MS MS (ESI) (ESI) m/z:355.1[M+1]*. m/z: 355.1[M+1]*.

[0577] Chemical

[0577] Formula: C ChemicalFormula: 19 H 2 2 N CHNO, 4 0 3 , Molecular Molecular Weight: Weight: 354.40. 354.40.

[0578] Step Step

[0578] 7b: Synthesis 7b: Synthesis of 3-(3'-oxo-6'-piperazin-1-yl-spiro[cyclopropane of 3-(3'-oxo-6'-piperazin-1-yl-spiro|cyclopropane-1,1' -1,1' isoindoline]-2'-yl)piperidine-2,6-dione fromthetheimide isoindoline|-2'-yl)piperidine-2,6-dione from imide product product of step of step 6 6

O O NH NH 0 0 o N O HCI/Dioxane HCI/Dioxane O N O NH N N DCM N N N NH O DCM N o N HN HN Boc'.N Boc

[0579]

[0579] To aamixture To mixtureof of tert-butyl tert-butyl 4-[2'-(2,6-dioxo-3-piperidyl)-l'-oxo-spiro[cyclopropane 4-[2'-(2,6-dioxo-3-piperidyl)-1'-oxo-spiro[cyclopropane- -

1,3'-isoindoline]-5'-yl]piperazine-1-carboxylate 1,3'-isoindoline]-5'-yI]piperazine-1-carboxylate(90 (90mg, mg, 0.20 0.20 mmol, mmol, 11 eq) eq) in in dichloromethane (5 dichloromethane (5

mL)was mL) wasadded added hydrochloric hydrochloric acid acid (4 (4 M M in in dioxane, dioxane, 2.52.5 mL,mL, 50 eq) 50 eq) in one in one portion portion at at 2525 °C.°C. TheThe

mixture was mixture wasstirred stirred at at 25 25 °C °C for for 11 hour. hour.LCMS showed LCMS showed thethe product product waswas the the main main peak. peak. The The mixture was mixture wasconcentrated concentratedininvacuum. vacuum.TheThe crude crude solid solid TheThe product product 3-(3'-oxo-6'-piperazin-1-yl 3-(3'-oxo-6'-piperazin-1-yl-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl)piperidine-2,6-dione spiro[cyclopropane-1,1'-isoindoline]-2'-yl)piperidine-2,6-dione (70 (70 mg, 0.18 mmol, mg, 0.18 90% mmol, 90% yield, yield,

hydrochloride) was hydrochloride) obtainedasasa abrown wasobtained brownsolid, solid,which whichwas was usedinto directlyused directly intothe thenext nextstep step without without further purification. further purification.

[0580] LCMS:

[0580] LCMS: MS MS (ESI) (ESI) m/z:355.1[M+1] m/z: 355.1[M+1]*.

248

Formula: C ChemicalFormula:

[0581] Chemical

[0581] 19 H 2 2 N CHNO, 4 0 3 , Molecular Molecular Weight: Weight: 354.40 354.40

[0582] Step Step

[0582] 8: Synthesis 8: Synthesis of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-

cyclobutyl]-4-[4-[[4-[2'-(2,6-dioxo-3-piperidyl)-1'-oxo-spiro[cyclopropane-1,3'-isoindoline] cyclobutyl|-4-[4-[[4-[2'-(2,6-dioxo-3-piperidyl)-1'-oxo-spirolcyclopropane-13'-isoindolinq]-

5'-yl]piperazin-1-yl]methyl]-1-piperidyl]benzamide 5'-yl|piperazin-1-yl|methyl]-1-piperidyl|benzamide

NH

HN .0CI CI 0 0 0

N HN, NH 2023248067 N NH, NH, N N 0NH N

Hold CI NaBH(AcO), NaBH(AcO)TEA, DCE, , TEA, 3 30 °C, DCE, 12.512.5h 30 °C, h N N N

[0583] To a To

[0583] a solution solution of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4 of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-

(4- (4- formyl-1-piperidyl)benzamide (63mg, formyl-1-piperidyl)benzamide (63 mg,0.12 0.12mmol, mmol, 1 eq) 1 eq) in in 1,2-dichloroethane 1,2-dichloroethane (3 (3 mL)mL) was was

added triethylamine (38 added triethylamine (38 mg, mg,0.38 0.38mmol, mmol,3 eq) 3 eq)and and 3-(3'-oxo-6'-piperazin-1-yl 3-(3'-oxo-6'-piperazin-1-yl-

spiro[cyclopropane-1,1'-isoindoline]-2'-yl)piperidine-2,6-dione (50 spiro[cyclopropane-1,1'-isoindoline]-2-yl)piperidine-2,6-dione (50 mg, mg,0.12 0.12mmol, mmol, 1 eq, 1 eq,

hydrochloride). The mixture hydrochloride). The mixturewas wasstirred stirred at at 30°C for 30 30°C for 30 min. min. Sodium Sodiumtriacetoxyborohydride triacetoxyborohydride (54(54

mg, 0.25 mg, 0.25 mmol, mmol,2 2eq) eq)was wasadded, added,then thenthethemixture mixturewaswas stirredatat30°C stirred 30°Cfor for1212hours. hours.LCMS LCMS showed thereaction showed the reaction was wascompleted completedandand desired desired MS MS can can be detected. be detected. The The reaction reaction mixture mixture was was

concentrated under reduced concentrated under reducedpressure pressuretotoremove removesolution. solution.The Theresidue residuewas was purifiedbybySemi- purified Semi preparative reverse preparative phase reverse HPLC phase HPLC(column: Phenomenex (column: PhenomenexSynergi SynergiC18 C18 150*25*Oum;mobile 150*25*10um;mobile

phase: [water(0.225%FA)-ACN];B%: phase: 40%-70%,10min)

[water(0.225%FA)-ACN];B%: 40%-70%,10min) to to giveN-[3-(3-chloro-4-cyano- give N-[3-(3-chloro-4-cyano phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-[4-[[4-[2'-(2,6-dioxo-3-piperidyl)-'-oxo phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-[4-[4-[2'-(2,6-dioxo-3-piperidyl)-1'-oxo-

spiro[cyclopropane-1,3'-isoindoline]-5'-yl]piperazin-1-yl]methyl]-1-piperidyl]benzamide (17.8 spiro[cyclopropane-1,3'-isoindoline]-5'-yl]piperazin-1-yl]methyl]-1-piperidylJbenzamide(17.8

mg, 0.02 mg, 0.02 mmol, mmol,16%16% yield, yield, 98%98% purity) purity) as as a white a white solid. solid.

[0584] LCMS:

[0584] MS MS LCMS: (ESI) (ESI) m/z:932.3 m/z: 932.3 [M+1].

[M+1]*.

[0585] ¹H NMR:

[0585] 1H NMR: (400MHz, (400MHz, DMSO-d) DMSO-d) :10.88 : 10.88 (s, 1H),(s,8.22 1H),(s, 8.221H), (s, 1H), 7.917.91 (d,(d, J=8.8 J=8.8Hz, Hz, 1H), 1H), 7.74 7.74 (d, (d, J=8.8 J=8.8 Hz, Hz, 2H), 7.53 -7.45 2H), 7.53 7.45(m, (m,2H), 2H),7.21 7.21(d, (d, J=2.4 J=2.4Hz, Hz,1H), 1H),6.99 6.99(dd, (dd,J=9.2, J=9.2, 17.6 17.6 Hz, 4H), 4H), 6.73 6.73 (s, (s, 1H), 1H), 4.33 4.33 (s, (s,1H), 1H),4.06 4.06(d, (d,J=9.2 J=9.2Hz, Hz,1H), 1H),3.86 3.86(d, J=12.4 (d, J=12.4Hz, Hz,3H), 3.32- 3.29 3H),3.32 3.29 (m, 9H), 2.80 (m, 9H), 2.80 (t, (t, J=12.0 J=12.0 Hz, 3H), 2.59 -2.54 Hz, 3H), 2.54(m, (m,4H), 4H),2.22 2.22(d, (d,J=6.8 J=6.8Hz, Hz,2H), 2H),1.81 1.81(d, (d,J=10.3 J=10.3Hz, Hz, 4H), 1.55 --1.47 4H), 1.55- 1.47(m, (m,2H), 1.45- 1.31 2H), 1.45 1.31 (m, (m, 2H), 2H), 1.25 - 1.17(s,(s, 8H), 1.25 1.17 8H),1.13 1.13(s, (s, 6H). 6H).

[0586] Chemical

[0586] Formula: C ChemicalFormula: CHCINO, 70, Molecular 4 7 H 5 4 ClNMolecular Weight: Weight: 832.43. 832.43.

Total

[0587] Total

[0587] H count H count from from HNMR HNMR data: 54.54. data:

[0588] C. C.

[0588] Exemplary Exemplary Synthetic Synthetic Schemes Schemes for for Exemplary Exemplary Androgen Androgen Receptor Receptor Binding Binding

Moiety Based Compounds Moiety Based Compounds that that areareImide ImideIsosteres Isosteres

249

[0589] General

[0589] General Synthetic Scheme SyntheticScheme C-1 C-1

[0590] Synthesis

[0590] Synthesis of building of building blockblock N-((r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide

CI CI0" O,, 0 O NC N N N N: HH 2023248067 NN N N NH NH

[0591]

[0591] Synthetic Scheme Synthetic Scheme NH 2HCI NHHCI Bac Boc N N N N N B On N 0 C1 CI 0 o o Boc 0 o Boc' N N HO Ho I N N oO NH NC HO NO NN NH NH Ho N cCI DMA, DIPEA,130130 DMA,DIPEA, °C,°C, o/n o/n N' HATU, DIPEA, DCM HATU, DIPEA, DCM CI 5 ~Bac N Boc rt, o/n rt, o/n 0 "O 65% 86% 86%

CI CI CI CN CN

C1 CI O,, O O 0 IZ N N NC H HCI, HCI, 1,4-dioxanme 1,4-dioxanme N N rt,4 hNH.HCI rt, 4h NH.HCI 100% 100%

[0592]

[0592] Step1:1:Synthesis Step Synthesisof of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid acid 0 o Boc O N Boc,N O H NH Ho HO N N HO Ho N N N N DMA, DIPEA,130 DMA, DIPEA, 130°C, °C, o/n o/n N, N, CI CI DMA, 65% Boc 65%

[0593]

[0593] 6-Chloronicotinic (1.6 g, acid (1.6 6-Chloronicotinic acid 10.0 mmol) g, 10.0 mmol) was dissolvedininN,N-dimethylacetamide wasdissolved N,N-dimethylacetamide (15 (15 mL), andtert-butyl mL), and tert-butyl piperazine-1-carboxylate (1.9 g, piperazine-1-carboxylate (1.9 g, 10.0 10.0 mmol) andethyldiisopropylamine mmol) and ethyldiisopropylamine (2.6 (2.6 g, g,20 20 mmol) wereadded mmol) were addedthereto, thereto, followed followedbybystirring stirring at at 130° 130 CC overnight. overnight. The The reaction reaction mixture was mixture wasconcentrated concentratedunder underreduced reduced pressure,and pressure, and totothe theobtained obtainedresidue residuewas wasadded added a 1a M 1 M aqueous NaOH aqueous NaOH solution solution (10(10 mL), mL), followed followed by washing by washing with with CHCl CHC13 (50ThemL). (50 mL). The pH of pH of the the

aqueous layer was aqueous layer wasadjusted adjustedtoto around around6 6toto 77 by by the the addition addition of of 11 M hydrochloric acid, M hydrochloric acid, followed followed

250 by extraction extraction with CHC13 CHCl (50(50 mLmL x 3). TheThe organic layer waswas dried over anhydrous sodium 2023248067 10 Oct 2023 by x 3). organic layer dried over anhydrous sodium sulfate sulfate and and the the solvent solvent was was concentrated concentrated under reduced pressure. under reduced pressure. The Theobtained obtainedresidue residuewas was purified by purified by silica silicagel gelcolumn column chromatography (CH 2 C 2/MeOH chromatography (CHCl/MeOH = 10/1)=to10/1) give to6-(4-(tert- give 6-(4-(tert butoxycarbonyl)piperazin-1-yl)nicotinic acid(2.0 butoxycarbonyl)piperazin-1-yl)nicotinic acid (2.0 g, g, 65 65%% yield)asasaa white yield) white solid. solid.

[0594] LC-MS

[0594] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile 95%from 95% Phase: Phase: from

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed 95% changedtoto95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min). min). Purity is Purity is 83.17%,RtRt==1.312 83.17%, 1.312min; min;MSMS Calcd.: Calcd.: 307.15; 307.15; MS MS Found: Found: 308.2308.2 [M+H]*.

[M+H].

[0595] Chemical

[0595] Formula: C ChemicalFormula: 1 5 H 2 1N CHNO, 3 0 4 , Molecular Molecular Weight: Weight: 307.34. 307.34.

[0596] Step Step

[0596] 2: Synthesis 2: Synthesis of tert-butyl of tert-butyl 4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate

NH 2 HCI NH2HCI

OII

0 C CI o HO N N NC CI Ho CI 0, O,, O NC O N N N N IZ N N N N, N, HATU, HATU, DIPEA, DCM DIPEA, DCM NC H Boc rt, o/n rt, o/n H Boc 86% 86% N N N Boc

[0597] A mixture

[0597] A mixture of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid acid (614 (614 mg, mg, 2.0 2.0 mmol),,4-(1r,3r)-3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-chlorobenzonitrile mmol), 4-((1r,3r)-3-amino-2,2,4,4-tetramethylcyclobutoxy)-2-chlorobenzonitrilehydrochloride hydrochloride (630 mg, 2.0 (630 mg, 2.0 mmol), mmol),2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (1.1 g,g, 3.0 hexafluorophosphate (1.1 3.0 mmol) mmol)and and ethyldiisopropylamine ethyldiisopropylamine (516 (516 mg, mg, 4.0 4.0 mmol) mmol) in in dichloromethane (20mL) dichloromethane (20 mL) was was stirredatatroom stirred room temperature temperature overnight. overnight. Water Water (50 (50 mL) mL) was added was added

and extracted with and extracted dichloromethane(50 with dichloromethane (50mLmL x 3).Combined X 3). Combined organic organic layers layers werewere washed washed by brine by brine

(50 mL 2), dried mL x x2), dried over over anhydrous anhydroussodium sodium sulfate.The sulfate. Thesolvent solventwas wasconcentrated concentrated to to givethe give the residue, which residue, waspurified which was purified by by column columnchromatography chromatography on silica on silica gelgel (petroleum (petroleum ether/ethyl ether/ethyl

acetate 1/1)totogive acetate ==1/1) givetert-butyl tert-butyl4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4- 4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate (977 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate (977 mg,mg, 86% 86 % yield) yield) as as aa white solid. white solid.

251

Waters X-Bridge (50 mm*4.6 C18mm*4.6 2023248067 10 Oct 2023

[0598] LC-MS

[0598] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters X-Bridge C18 (50

mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min). min). Purity is Purity is 88.26%,RtRt==2.161 88.26%, 2.161min; min;MSMS Calcd.: Calcd.: 567.26; 567.26; MS MS Found: Found: 568.3568.3 [M+H]*.

[M+H].

[0599]

[0599] ¹H 'H NMR NMR (400 MHz, (400 DMSO-d) MHz, 81.12 (6H, DMSO-d) s),(6H, 61.12 1.22s), 1.22 (6H, (6H, s), s),(9H, 1.43 (9H, 1.43s), s), 3.42-3.44 3.42-3.44

(4H, m),3.60-3.63 (4H, m), 3.60-3.63 (4H, (4H, m), 4.02-4.07 m), 4.02-4.07 (1H, (1H, m), 4.31m), 4.31 (1H, s), (1H, s), 6.88 6.88 (1H, d, J (1H, = 8.8 d, J=7.00 Hz), 8.8 (1H, Hz), 7.00 (1H, dd, dd, JJ= 8.4, 2.4 = 8.4, 2.4 Hz), Hz),7.21 7.21(1H, (1H, d, Jd,=J= 2.4 2.4 Hz),Hz), 7.65 7.65 (1H, (1H, d, J =d, J= 9.2 9.27.91 Hz), Hz),(1H, 7.91 d, (1H, d, Hz), J = 8.8 J= 8.8 Hz), 7.99 (1H,dd, 7.99 (1H, dd,J J= 8.8,2.42.4 = 8.8, Hz), Hz), 8.648.64 (1 d,H,J d, (1 H, = 2.4 2.4 Hz). J=Hz).

[0600] Chemical

[0600] Formula: C ChemicalFormula: 30 H 3 8 ClN CHCINO, 5 04 , Molecular Molecular Weight: Weight: 568.11. 568.11. Total

[0601] Total

[0601] H count H count from from HNMR HNMR data: 38.38. data:

[0602] Step Step

[0602] 3: Synthesis 3: Synthesis of N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide hydrochloride tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide hydrochloride

CI CI 00, O,, CI O,, CI o0 NC NC N I2 NN HCI, 1,4-dioxane HCI, 1,4-dioxane NC N IZ N N NC N N -N rt, 4 hh rt, N N N NBoc Boc 100% 100% NH.HCI NH.HCI

[0603] A mixture

[0603] A mixture of tert-butyl of tert-butyl 4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl carbamoyl)pyridin-2-yl)piperazine-1-carboxylate(405 tetramethylcyclobutyl carbamoyl)pyridin-2-yl)piperazine-1-carboxylate (405 mg,mg, 0.7 0.7 mmol) mmol) in in

HCl/1,4-dioxane(10 HCI/1,4-dioxane (10mL) mL)waswas stirredatatroom stirred room temperature temperature forfor 4 h.The 4 h. The solvent solvent was was removed removed in in vacuum vacuum totogive giveN-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6- N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6 (piperazin-1-yl)nicotinamide hydrochloride hydrochloride(353 (353mg, mg,100% 100yield) % yield) as aaswhite a white solid. solid.

[0604] LC-MS

[0604] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min). min). Rt = 1.791Rt = 1.791 min; MS min; Calcd.:467.21; MS MS Calcd.:467.21; MS Found: Found: 468.3 468.3 [M+H]*.

[M+H].

[0605] Chemical

[0605] Formula: C ChemicalFormula: 2 5 H 3 1 Cl 2 N CHClNO, 5 02 , Molecular Molecular Weight: Weight: 504.45 504.45 General

[0606] General

[0606] Synthetic Scheme SyntheticScheme C-2 C-2

Synthesis

[0607] Synthesis

[0607] of building of building block block tert-butyl tert-butyl 4-(4-formylpiperidin-1-yl)benzoate 4-(4-formylpiperidin-1-yl)benzoate

252

Oct 2023

NNO O

[0608]

[0608] Synthetic scheme: Synthetic scheme: 2023248067 10 OH OH OHNo OH F OF HN HNO' HN H, ONN

O DMSO,120 DMSO, 120°C, 91.2% o/n °C,o/n >r O o 91.2%

DMP, DCM, DMP, DCM, 0 O o N rt,1h rt, 1 h N O0 81% O 81%

[0609]

[0609] Step 1: Step Synthesis of 1: Synthesis tert-butyl4-(4-(hydroxymethyl)piperidin-1-yl)benzoate of tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate

OH OH OHNo OH F N HNOH HN O O O DMSO,120 DMSO, 120°C, °C,o/n o/n O 91.2% 91.2% O 0

[0610] To a To

[0610] a solution solution of tert-butyl of tert-butyl 4-fluorobenzoate (23 (23 4-fluorobenzoate g, 0.12 g, 0.12 mmol) mmol) in DMSO in DMSO (100 (100 mL) mL) was added was addedpiperidin-4-ylmethanol piperidin-4-ylmethanol(40.5 0.35mmol). (40.5g,g,0.35 mmol).TheThe mixture mixture was was heated heated to 120 to 120 °C °C overnight under nitrogen. overnight under nitrogen. After After cooling cooling to to room temperature, water room temperature, water(50 (50mL) mL)was was added added to to thethe

reaction mixture, reaction mixture, and extracted with and extracted with ethyl ethyl acetate acetate(20 (20mL 3). The x 3). mL X The organic layer layer was washed was washed

with with brine (15 (15 mL 3). The mL xx3). The combined combined organic organic phases phases were were dried dried over over anhydrous anhydrous sodium sodium sulfate sulfate

and concentrated in and concentrated in vacuo, vacuo, and and purified purified by by CC CC(PE/EA (PE/EA = 10:1) = 10:1) to to give give compound compound tert-butyl tert-butyl 4-(4 4-(4-

(hydroxymethyl)piperidin-1-yl)benzoate (31g,91.2%) (hydroxymethyl)piperidin-1-yl)benzoate (31g, 91.2%) as as a white a white solid. solid.

LCMS

[0611] LCMS

[0611] (Agilent (Agilent LCMSLCMS 1200-6120, 1200-6120, Column: WatersWaters Column: X-Bridge X-Bridge C18 (50 4.6 xmm4.6 C18mm(50x mm mm x 3.5 X m); Column 3.5 µm); ColumnTemperature: Temperature: 40 40 °C; °C; Flow Flow Rate: Rate: 2.0 2.0 mL/min; mL/min; Mobile Mobile Phase:Phase: from from 90% 90%

[(total 10mM

[(total 10mM AcONH AcONH4)4)water/CHCN=900/100 water/CH3CN=900/100 (v/v)] (v/v)] andand10%10% [(total 10mM

[(total 10mM AcONH 4 AcONH4) )

water/CH3CN=100/900 (v/v)]toto 10% water/CHCN=100/900 (v/v)] 10%[(total

[(total 10mM AcONH water 10mM AcONH4) 4) water /CH 3 CN=900/100 /CHCN=900/100 (v/v)] (v/v)]

and 90%[(total and 90% [(total 10mM 10mM AcONH AcONH4) 4) water/CH3CN=100/900 water/CH3CN=100/900 (v/v)] in (v/v)] in 1.6 1.6 min, thenmin, then under under this this

253 condition for 2.4 2.4 min, min, finally finallychanged changed to to 90% [(total10mM AcONH 4) water/CH3CN=900/100 2023248067 2023 condition for 90% [(total 10mM AcONH4) water/CHCN=900/100

(v/v)] (v/v)] and and 10% [(total10mM 10% [(total AcONH 10mM AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] in (v/v)] 0.1 minin and 0.1 under min and under this this

10 Oct condition for 0.7 condition for 0.7 min). min). Purity Purity isis99.57%, 99.57%, Rt Rt == 2.035 min.; MS 2.035 min.; Calcd.: 291.2; MS Calcd.: MSFound: 291.2; MS Found: 292.2 292.2

[M+H]+.

[M+H]+.

HPLC

[0612] HPLC

[0612] (Agilent (Agilent HPLC 1200,1200, HPLC Column: Column: Waters Waters X-Bridge X-Bridge C18 (150 mm Xmm C18 (150 4.6xmm 4.6xmm 3.5x 3.5 pm); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 1.0 mL/min; 1.0 mL/min; MobileMobile Phase:Phase: from from 95% 95%+[water

[water 10 + 10 mMNH4HCO] mM NH 4HCOand 5%5% 3] and [CH 3 CN]

[CHCN] to 0% to 0% [water++10

[water 10 mM mMNH4HCO] NH 4HCOand 3] and 100% 100% [CH 3 CN]

[CHCN] in 10 in 10 min, then min, then under under this this condition condition for for 55 min, min, finally finallychanged changed to to95% [water ++ 10 95% [water 10 mM mM NH 4HCO 3] NH4HCO]

and 5%[CHCN] and 5% [CH 3 CN] in 0.1 in 0.1 min min and under and under this this condition condition for for 5 min). 5 min). Purity Purity is 93.27%, is 93.27%, Rt =Rt9.542 = 9.542

min. min.

[0613]

[0613] ¹H NMR (400 (400 'H NMR MHz,MHz, CDCl)CDC 1.29-1.40 (2H, m), 1.49 (1H, d, J = 5.4 Hz), 1.57 (9H, 3) 6 1.29-1.40 (2H, m), 1.49 (1H, d, J= 5.4 Hz), 1.57 (9H,

s), s), 1.70-1.75 (1H,m),m), 1.70-1.75 (1H, 1.82 1.82 (2H,(2H, d, J d, = J= 12.8 12.8 Hz), Hz), 2.80-2.87 2.80-2.87 (2H, m),(2H, 3.53 m), (2H,3.53 t, J (2H, = 5.8 t,Hz), J= 5.8 Hz), 3.87-3.90 (2H, 3.87-3.90 (2H, m),m), 6.85 6.85 (2H,(2H, d, J d, J=Hz), = 9.2 9.2 7.84 Hz), (2H, 7.84d,(2H, d, J= J = 9.2 Hz).9.2 Hz).

Formula: C ChemicalFormula:

[0614] Chemical

[0614] 17 H 2 5 NO CHNO, 3 , Molecular Molecular Weight: 291.39. Weight:291.39.

[0615]

[0615] Total H Total H count HNMR fromHNMR count from data: 25. 25. data:

[0616]

[0616] Step 2: Step Synthesisof of 2: Synthesis tert-butyl tert-butyl 4-(4-formylpiperidin-1-yl)benzoate 4-(4-formylpiperidin-1-yl)benzoate

N OH OH DMP, DCM, DMP, DCM, 0 O N O N N O rt,l rt, 1h1 h N

81% O / 81%

[0617] To a To

[0617] a solution solution of tert-butyl of tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (300 (300 mg, mg, 1.03 1.03 mmol) mmol) inindichloromethane dichloromethane(20(20 mL) mL) waswas added added Dess-Martin Dess-Martin periodinane periodinane (1.31 (1.31 g, 3.09 g, 3.09

mmol)slowly mmol) slowlyatat0 0°C. °C. The Thereaction reactionmixture mixturewas wasstirred stirredatat room roomtemperature temperaturefor for1 1h.h. Then Then filtered, filtered,and andconcentrated concentrated in invacuo vacuo to togive givecompound tert-butyl 4-(4-formylpiperidin-1 compound tert-butyl 4-(4-formylpiperidin-1-

yl)benzoate (240 yl)benzoate (240 mg, mg,81%) 81%)as as a apale paleyellow yellowsolid. solid.

[0618] Exemplary

[0618] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 46 Compound 46

254

H ZI H O 0NO N 0 O N N O . N N N H _jr ZI

N ~ N N 2023248067 CI CI 0" "O 0 O

N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-((2-(2,6 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-(2-(2.,6-

dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)pentyl)piperazin-1 dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)oxy)penty)piperazin-1-

yl)nicotinamide yl)nicotinamide

106191

[0619] Synthetic Scheme Synthetic Scheme

a0 o B o 00 0 0 O ó o o OH HSO(cat.), MeOH N OH H 2 S04 (Cat.).MeOH_ o CsCO, Pd(PhP) OsO, NaIO o 90 °C, 16 h 0 Cs2 CO3 , Pd(Ph 3P)4 N o Br 04 NaIO 4 o Br Oe arDMF/HO=10/1 MF/H0=10/ 0 o 0MeCN:acetone:H,0(1:1:1) 0 MeCN:acetone:HO(1:1:1) o 100 °C, 4h 91% 91% 100 OC,4h N.r.t, r.t, 4h 4h '0 100% 35% o 100% 35%

0 o HN HN 0 o 0 o o SO CI NH 0 IH N-: 2 o HCI HCI H o"P HN o NN HN 0 NO ZI O N0 O O o N o BBr, o TEA, AcOH, TEA, AcOH, NaBH 3 CN NaBHCN 0 o B___,DCM, -78 °C to rt. D___-78_Ctort N KCO, DMF, 40 °C, o/n MeOH, 0~rt, o/n NeH jt:' NI / o/n I-t.n K 2C0 3, DMF, 40OC,a/n HO 23% 0 59% -0o -138% 38% HO o 59%

NH CI

HN N N N N NNN N 'N :TN N CI ci ~o'~ 0 O Ó HN o o HN

___ ___ ___ H0r CI N o OIEAKI, DIEA, CHCN,100 OC KI, CHCN, 100 °C / 0" $/~ -/N\'-0 N N o tube sealed tube sealed 0 N0N N o 8% 8%

106201 Step

[0620] Step 1: 1: Synthesis of Synthesis of methyl methyl2-bromo-4-methoxybenzoate 2-bromo-4-methoxybenzoate

255

0 0 Oct 2023

0 ) 'OHOH H 2SO 4 (cat.), MeOH HSO(cat.), MeOH O

Br Br 90 C, 1616hh 90 °C, O Br Br

91% 91% 2023248067 10

[0621] To a To

[0621] a solution solution of 2-bromo-4-methoxybenzoic of 2-bromo-4-methoxybenzoic acid acid (5.0 g, (5.0 21.7g,mmol) 21.7 in mmol) in methanol methanol (50 (50 mL)was mL) wasadded added 98% 98% sulfuric sulfuric acid acid ml).The (0.5ml). (0.5 The reactionmixture reaction mixture waswas heated to to heated 90 90 °C °C forfor 16 16 h h under nitrogen under nitrogen gas, gas, and and concentraction concentraction under underreduced reducedpressure. pressure. After Aftercooling coolingtoto room room temperature, sodiumbicarbonate temperature, sodium bicarbonate(2.0 (2.0M)M)was was added added to to adjust adjust PH=8. PH=8. Thus Thus was was extracted extracted withwith

ethyl ethyl acetate acetate (50 (50 mL 3). The mL xx 3). organic layer The organic layer was washedwith was washed withbrine brine(30 (30mL). mL).The The combined combined

organic phases were organic phases weredried dried over over anhydrous anhydroussodium sodium sulfate,filtered, sulfate, filtered, and and concentrated concentrated in in vacuo vacuototo give give 2-bromo-4-methoxybenzoate 2-bromo-4-methoxybenzoate (4.8(4.8 g, 91%) g, 91%) as yellow as yellow oil.oil.

Agilent

[0622] Agilent

[0622] LCMS LCMS 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge C18 C18 X-Bridge (50 xmm (50 mm 4.6 xmm 4.6x mm 3.5x 3.5 ptm); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 2.0 mL/min; 2.0 mL/min; MobileMobile Phase:Phase: from from 90% 90%

[(total [(total 10mM AcONH4) 10mM AcONH4) water/CH3CN=900/100 water/CHCN=900/100 (v/v)](v/v)] and and 10% 10% [(total

[(total 10mM 10mM AcONH4) AcONH4)

water/CH3CN=100/900 (v/v)]toto 10% water/CHCN=100/900 (v/v)] 10%[(total

[(total 10mM AcONH water 10mM AcONH4) 4) water/CHCN=900/100 /CH 3 CN=900/100 (v/v)] (v/v)]

and 90%[(total and 90% [(total 10mM AcONH 10mM AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] (v/v)] in 1.6 in then min, 1.6 min, underthen under this this

condition for 2.4 condition for 2.4 min, min, finally finallychanged changed to to 90% [(total10mM 90% [(total AcONHwater/CHCN=900/100 10mM AcONH4) 4) water/CH3CN=900/100

(v/v)] (v/v)] and and 10% [(total 10mM 10% [(total AcONH4) 10mM AcONH4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] in (v/v)] 0.1 minin and min and 0.1 under under this this

condition for 0.7 condition for 0.7 min. min. Purity Purity isis98.94%, 98.94%, Rt Rt == 2.609 2.609 min; min; MS Cald.:243.97; MS Calcd.: 243.97;MS MS Found: Found: 245.0 245.0

[M+H]*.

[M+H].

[0623] Step Step

[0623] 2: Synthesis 2: Synthesis of methyl of methyl 2-allyl-4-methoxybenzoate 2-allyl-4-methoxybenzoate

O0'BB 0 O 01 0 O O 0 CsCO, Pd(Ph 3P)4 Cs 2C 3, Pd(PhP) O O0 C Br Br DMF/H 2 0=10/1 DMF/HO=10/1 0 o 100 °C, 100 4 h °C, 4h

100% 100%

[0624] To a To

[0624] a solution solution of methyl of methyl 2-bromo-4-methoxybenzoate 2-bromo-4-methoxybenzoate (3.0 g, (3.0 g, 12.3 12.3 cesium mmol), mmol), cesium carbonate (12.0 g, carbonate (12.0 g, 36.9 36.9 mmol), 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.98 mmol), 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.98 g, g, 18.5 18.5

256

mmol)ininV,N-dimethylformamide mmol) N,N-dimethylformamide /water /water (30.0(30.0 mL/3.0 mL/3.0 mL) was was mL)added added tetrakis(triphenylphosphine)palladium (1.42 tetrakis(triphenylphosphine)palladium (1.42 g,g, 1.23 1.23 mmol) mmol)under undernitrogen nitrogenatmosphere. atmosphere. TheThe

reaction mixture was heated to 100 °C and stirred for 4 h. The resulting reaction was reaction mixture was heated to 100 °C and stirred for 4 h. The resulting reaction was

concentrated under concentrated underreduced reducedpressure, pressure, and andthen thenwater water(10 (10mL) mL)waswas added. added. TheThe mixture mixture was was extracted extracted with with ethyl ethyl acetate acetate (50 (50mL 3). The x 3). mL x The combined organicphase combined organic phasewas waswashed washed with with brine brine (20(20

mL), dried mL), dried over over anhydrous anhydroussodium sodium sulfate,filtered, sulfate, filtered, and concentrated. The and concentrated. The residue residue was waspurified purified by silica gel chromatography column (petroether/ethyl acetate = 4:1) to give the methyl 2-allyl by silica gel chromatography column (petroether/ethyl acetate = 4:1) to give the methyl 2-allyl-

4-methoxybenzoate 4-methoxybenzoate (2.6g,g,100%) (2.6 100%) as as yellow yellow oil. oil.

[0625] Agilent

[0625] Agilent LCMS LCMS 1200-6110, 1200-6110, Column: Column: Waters Waters X-Bridge (50 (50 C18 C18 X-Bridge mm xmm x 4.6 4.6 mm xmm x 3.5 3.5 ptm); Column µm); Column Temperature: Temperature: 40 40 °C; °C; Flow Flow Rate: Rate: 1.5 mL/min; 1.5 mL/min; Mobile Mobile Phase:Phase: from from 95% 95% +[water

[water

+ 0.05% TFA] and 0.05% TFA] and 5% 5%[CHCN

[CH 3CN + 0.05% + 0.05% TFA]TFA] to 0% to 0% [water

[water + 0.05% + 0.05% TFA] TFA] andand 100% 100% [CH+ 3CN

[CHCN

+ 0.05 TFA] 0.05 % TFA] in 1.5 in 1.5 min, min, then then under under thisthis condition condition forfor 0.50.5 min,finally min, finallychanged changedto to 95% 95% [water

[water +

+ 0.05% TFA]andand 0.05% TFA] 5% 5% [CH

[CHCN CN + TFA] + 30.05% 0.05% in TFA] 0.1 min in and 0.1 min underand under this this condition condition for 0.5 min. for 0.5 min.

Purity is Purity is 96.85%, Rt == 1.293 96.85%, Rt min; MS 1.293 min; MSCalcd.: Calcd.:206.09; 206.09;MSMS Found:207.3 Found:207.3 [M+H]*.

[M+H].

[0626] Step Step

[0626] 3: Synthesis 3: Synthesis of methyl of methyl 4-methoxy-2-(2-oxoethyl)benzoate 4-methoxy-2-(2-oxoethyl)benzoate

0 0 O 0 O OsO,4 , NalO Os0 NalO 4 O 0 MeCN:acetone:H 2 0(1:1:1) MeCN:acetone:HO(1:1:1) >NO I rt, 4 h rt,4h

35% 35% 0

[0627] To a To

[0627] a solution solution of methyl of methyl 2-allyl-4-methoxybenzoate (1.20 (1.20 2-allyl-4-methoxybenzoate g, 5.83 5.83 mmol) g, mmol) and and osmium osmium tetraoxide (5 tetraoxide (5 mg) in acetonitrile, mg) in acetonitrile, acetone, and acetone, andwater water(v: v: v: (v: v=10 v=10mL: mL: 10 10 mL: 10 mL) mL: 10 mL)was wasadded added sodium periodate (4.99 sodium periodate (4.99 g, g, 23.3 23.3 mmol) mmol)atat00°C. °C. The Themixture mixturewas was stirredatat room stirred roomtemperature temperatureforfor4 4 h. The mixture was filtered through a pad of celite and extracted with ethyl acetate (20 x 3 mL). h. The mixture was filtered through a pad of celite and extracted with ethyl acetate (20 x 3 mL).

The organic The organic layer layer was was separated, separated, washed washedwith withwater waterandand brine,dried brine, driedover overanhydrous anhydrous sodium sodium

sulfate, filtered, and sulfate, filtered, concentrated.TheThe and concentrated. residue residue was purified was purified by prep-TLC by prep-TLC (petroether/ethyl (petroether/ethyl

acetate acetate == 4:1) 4:1) to togive givecompound methyl4-methoxy-2-(2-oxoethyl)benzoate compound methyl 4-methoxy-2-(2-oxoethyl)benzoate (420 (420 mg, as mg, 35%) 35%) as yellow oil. yellow oil.

LC-MS

[0628] LC-MS

[0628] LCMSLCMS (Agilent (Agilent 1200-6110, 1200-6110, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50x mm C18 mm 4.6 x 4.6 mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 1.5 mL/min; 1.5 mL/min; Mobile Mobile Phase: Phase: from from 95% 95%

[water ++ 0.05%

[water 0.05% TFA] TFA] and and 5% 5% [CH 3CN

[CHCN + 0.05% + 0.05% TFA] TFA] to to 0%0% [water+ +0.05%

[water 0.05%TFA] TFA]andand100% 100%

[CH 3CN

[CHCN + 0.05 + 0.05 % TFA] % TFA] in min, in 1.5 1.5 min, then then this this under under condition condition for for 0.5 0.5 min,min, finally finally changed changed to to

257

Oct 2023 95%[water 95% 0.05%

[water+ +0.05% TFA] TFA] and and 5% [CH 5% [CHCN 3 CN + + 0.05% 0.05% TFA] in 0.1 mininand TFA] min and 0.1under thisunder this condition condition

for for 0.5 0.5 min.). min.).Purity Purityisis 96.26%, 96.26%,Rt Rt==1.007 1.007 min; min; MS Calcd.: 208.1; MS Calcd.: 208.1; MS MSFound: Found: 209.3 209.3 [M+H]*.

[M+H].

[0629]

[0629] Step 4: Step Synthesis of 4: Synthesis of 3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine 3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-

2,6-dione 2,6-dione 2023248067 10

O NH O O H 2N O HCI HCI O HN

HN O o O TEA, AcOH, TEA, AcOH, NaBH 3 CN NaBHCN N N o MeOH,0~rt, MeOH, O-rt,o/n o/n N O 59% O 59%

[0630] To a To

[0630] a solution solution of methyl of methyl 4-methoxy-2-(2-oxoethyl)benzoate 4-methoxy-2-(2-oxoethyl)benzoate (420 mg, 2.02 (420 mmol) mg, 2.02inmmol) in methanol (6 mL) methanol (6 mL)was was added added a solution a solution of of 3-aminopiperidine-2,6-dione 3-aminopiperidine-2,6-dione hydrochloride hydrochloride (397(397 mg, mg,

2.42 mmol) 2.42 mmol)and andtriethylamine triethylamine(245 (245mg, mg, 2.24 2.24 mmol) mmol) in methanol in methanol (2 mL). (2 mL). The reaction The reaction mixture mixture

was stirred at was stirred atroom room temperature for 11 h, temperature for h, then then sodium cyanoborohydride(254 sodium cyanoborohydride (254 mg,mg, 4.04 4.04 mmol) mmol)

was addedatat 00 °C. was added °C. The reaction was The reaction was stirred stirred at atroom temperature overnight, room temperature overnight, water water (10 (10 mL) mL)was was added, and extracted added, and extracted with with ethyl ethyl acetate acetate (20 (20 mL 3), washed mL Xx 3), withwater washed with waterand andbrine, brine, dried dried over over anhydrous sodium anhydrous sodium sulfate,filtered, sulfate, filtered, and and concentrated. concentrated. The residue was The residue was purified purified by prep-TLC by prep-TLC

(dichloromethane/methanol (dichloromethane/methanol = = 20:1) 20:1) to to give3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H)- give 3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H) yl)piperidine-2,6-dione (340 yl)piperidine-2,6-dione (340 mg, 59%)asasa apale mg, 59%) paleyellow yellowsolid. solid.

LC-MS

[0631] LC-MS

[0631] LCMSLCMS (Agilent (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30X mm C18mm 3 mmx 3 mm x 2.5 x m); Column 2.5 µm); ColumnTemperature: Temperature: 40 40 °C;°C; Flow Flow Rate: Rate: 1.5 1.5 mL/min; mL/min; Mobile Mobile Phase:Phase: from from 95% 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN+ +10

[CHCN 10 mM mMNH4HCO] NH 4HCOto 3 ]to 5%5% [water ++ 10

[water 10 mM mM NH 4HCO NH4HCO] and 95%95% 3] and [CH+3 CN

[CHCN + 10 10 mM mM NH NH4HCO] in4HCO ]in 1.5 1.5 3min, min, then then this under undercondition this condition for for 0.50.5

min, finally min, finally changed changed 95%95% to to [water + 10+ mM

[water NHNH4HCO] 10 mM 4HCO 3] and and 5% 5%[CHCN

[CH 3 CN + 10 10 mM + mM NH 4 HCO 3] NH4HCO]

in 0.1 0.1 min min and and under this condition under this condition for for 0.5 0.5min.). min.).Purity Purityis is 80.84%, 80.84%,RtRt= =0.924 0.924min; min;MS Calcd.: MS Calcd.:

288.1; 288.1; MS MS Found: 289.1 289.1 [M+H]*.

[M+H].

[0632]

[0632] Step Synthesis of 5: Synthesis Step 5: of 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-

2,6-dione H ZI H H oo H N N 0 O 0o IN0 N O o BBr3 DCM, BBr, , DCM, -78-78 °C °C to rt. to rt. O 0- -~ N N o/n N oj[):tN 0/n OO 38% 38% Ho

258

[0633] To a To

[0633] a solution solution of 3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6 of 3-(6-methoxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6-

dione (220 mg, dione (220 mg,0.76 0.76 mmol) mmol)in in dichloromethane dichloromethane (10 (10 mL) mL) was added was added boron boron tribromide tribromide (0.5inmL) in (0.5 mL)

dichloromethane dichloromethane (2(2mL) mL) dropwise dropwise at at -78-78 °C °C andand stirredovernight stirred overnightatatroom room temperature.TheThe temperature.

reaction mixture reaction was added mixture was addedtotowater water(10 (10 mL) mL)andand sodium sodium bicarbonate bicarbonate (20 (20 mL),mL), thenthen extracted extracted

with dichloromethane/ with dichloromethane/methanol methanol (30 (30 mL mL x 5). X 5). TheThe organic organic layer layer waswas washed washed with with brinebrine (10 mL). (10 mL).

The combined The combined organic organic phases phases were were dried dried over over anhydrous anhydrous sodium sodium sulfate, sulfate, filtered, filtered, andand concentrated in vacuo. vacuo. The The residue residue was waspurified purified by byprep-TLC prep-TLC (dichloromethane/ methanol=10:1) 2023248067

concentrated in (dichloromethane/ methanol=10:1)

to give to give compound 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6-dione compound 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6-dione (80 (80 mg, 38%) mg, 38%)asasa ayellow yellowsolid. solid. LC-MS

[0634] LC-MS

[0634] LCMSLCMS (Agilent (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30x mm C18mm 3 mmx 3 mm x 2.5 2.5 pm); ColumnTemperature: µm); Column Temperature: 40 40 °C;°C; Flow Flow Rate: Rate: 1.5 1.5 mL/min; mL/min; Mobile Mobile Phase:Phase: from from 95% 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN+ +10

[CHCN 10 mM mMNH4HCO] NH 4 HCOto 3 ] to 5%5% [water ++ 10

[water 10mM mM NH 4 HCO NH4HCO] 3] and and 95%95% [CH+3CN

[CHCN + 10 10 mM mM NH NH4HCO] in4 HCO ]in 1.5 1.5 3min, min, then then this under undercondition this condition 0.50.5 for for

min, finally min, finally changed to to changed 95%95%

[water + 10+ mM

[water NHNH4HCO] 10 mM 4HCO 3] and and 5% 5%[CHCN

[CH 3CN + mM + 10 10mM NH 4 HCO 3] NH4HCO]

in 0.1 0.1 min min and and under this condition under this condition for for 0.5 0.5min.). min.).Purity Purityis is 96.22%, 96.22%,RtRt= =0.736 0.736min; min;MS Calcd.: MS Calcd.:

274.1; MS 274.1; MS Found: 275.1 275.1 [M+H]*.

[M+H].

[0635] Step Step

[0635] 6: Synthesis 6: Synthesis of 3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H) of 3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H)-

yl)piperidine-2,6-dione yl)piperidine-2,6-dione

H ZI H 0 N N 0 ZI H O O O O N N 0 0 O CI OTs CI -M OTs N N N 3 , DMF, K2 CODMF, KCO, 40 °C, 40 °C, o/noln

Ho O HO 23% 23% CI CI

[0636] To a To

[0636] a solution solution of 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6 of 3-(6-hydroxy-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-2,6-

dione (80 mg, dione (80 mg, 0.292 0.292 mmol) mmol)in in N,N-dimethylformamide N,N-dimethylformamide (5.0 was (5.0 mL) mL)added was added 5-chloropentyl 5-chloropentyl 4- 4 methylbenzenesulfonate (64.5mg, methylbenzenesulfonate (64.5 mg, 0.234 0.234 mmol) mmol) and and potassium potassium carbonate carbonate (121 0.876 (121 mg, mg, 0.876 mmol).mmol).

The mixture The mixturewas washeated heatedtoto4040°C°Covernight. overnight.After Aftercooling coolingtotort., rt., the thereaction reactionmixture mixturewas was added added

to water to water (10 (10 mL), and extracted mL), and extracted with with ethyl ethyl acetate acetate (20 (20 mL 3). The mL xx 3). organic layer The organic layer was was washed washed with brine with brine (10 (10 mL 3). The mL Xx3). The combined combined organic organic phases phases were were dried dried over over anhydrous anhydrous sodium sodium sulfate, sulfate,

filtered, filtered,and andconcentrated concentrated in invacuo. vacuo.The The residue residue was was purified purified by by prep-TLC (dichloromethane/ prep-TLC (dichloromethane/

259

Oct 2023 methanol=10:1) methanol=10:1) totogive give3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H)- 3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H) yl)piperidine-2,6-dione (25 yl)piperidine-2,6-dione (25 mg, 23%)asasa ayellow mg, 23%) yellowsolid. solid.

LC-MS

[0637] LC-MS

[0637] LCMSLCMS (Agilent (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30x mm C18mm 3 mmx 3 mm x 2.5 m); Column 2.5 µm); ColumnTemperature: Temperature: 40 40 °C;°C; Flow Flow Rate: Rate: 1.5 1.5 mL/min; mL/min; Mobile Mobile Phase:Phase: from 95% from 95% 2023248067 10

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN+ +10

[CHCN 10 mM mMNH4HCO] NH 4HCOto 3 ]to 5%5% [water ++ 10

[water 10 mM mM NH 4HCO NH4HCO] 3] and and 95%95% [CH+3 CN

[CHCN + 10 10 mM mM NH NH4HCO] in4HCO ]in 1.5 1.5 3min, min, then then this under undercondition this condition for for 0.50.5

min, finally min, finally changed to to changed 95% [water 95% + 10+ mM

[water NHNH4HCO] 10 mM 4HCO 3] and and 5% 5%[CHCN

[CH 3 CN + 10 10 mM + mM NH 4 HCO 3] NH4HCO]

in in 0.1 0.1 min min and and under this condition under this condition for for 0.5 0.5min.). min.).Purity Purityis is 93.68%, 93.68%,RtRt= =1.263 1.263min; min;MS Calcd.: MS Calcd.:

378.1; 378.1; MS MS Found: 379.1 379.1 [M+H]*.

[M+H].

[0638] Step Step

[0638] 7: Synthesis 7: Synthesis of N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-

6-yloxy)pentyl)piperazin-1-yl)nicotinamide 6-yloxy)pentyl)piperazin-1-yl)nicotinamide

NH N OHi H IZ NNC IZ N 0 N 0 0 N 0 N NNN

CI 0N N N N DIEA, KI, CHCN, 100 °C DIEA, KI, sealed tube CH3 CN, 100 °C CI NN H IZ N C 8% 8% N Nele H N Oubr-N CI CI 0

[0639] A solution

[0639] A solution of 3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H) of 3-(6-(5-chloropentyloxy)-1-oxo-3,4-dihydroisoquinolin-2(1H)-

yl)piperidine-2,6-dione (25 mg, yl)piperidine-2,6-dione 0.066 mmol) mg, 0.066 mmol)waswas dissolved dissolved in in acetonitrile(2(2 mL), acetonitrile mL),N-((1r,3r)-3- N-((1r,3r)-3 (3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide (31(31 (3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamidet

mg, 0.066 mg, 0.066 mmol), mmol),ethyldiisopropylamine ethyldiisopropylamine (17(17 mg,mg, 0.132 0.132 mmol), mmol), potassium potassium iodideiodide (2 was (2 mg) mg) was added to the added to the solution. solution. The The mixture mixture was heated to was heated to 100 100 °C °Cfor for 16 16 hh under undersealed sealed tube. tube. After After cooling cooling

to to rt., rt.,the the reaction reaction mixture was mixture was added added to water to water (10 and (10 mL), mL), and extracted extracted with with ethyl ethyl acetate (10acetate mL (10 mL x 3). 3). The The organic layer layer was was washed withbrine washed with brine(10 (10 mL mLX 3). x 3).The Thecombined combined organic organic phases phases werewere

dried overanhydrous dried over anhydrous sodium sodium sulfate, sulfate, filtered, filtered, and concentrated and concentrated in vacuo, in vacuo, then thenbypurified purified prep- by prep HPLCto togive HPLC givecompound compound N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-

6-yloxy)pentyl)piperazin-1-yl)nicotinamide (4.1 6-yloxy)pentyl)piperazin-1-yl)nicotinamide (4.1 mg, mg,8%) 8%) as as a whitesolid. a white solid. LC-MS

[0640] LC-MS

[0640] LCMSLCMS (Agilent (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 4.6 x 4.6 C18mm(50X mm mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

260

[CH 3CN] in 3.0 min, then under thiscondition condition for1.0 1.0min, min,finally finally changed changedtoto95% 95% [water + 10 2023248067 10 Oct 2023

[CHCN] in 3.0 min, then under this for [water + 10

mMNH4HCO] mM NH4 HCO ] and and 35% 5% in

[CHCN] [CH 3CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.). min.).isPurity Purity is 2.923min; 87.84%,RtRt==2.923 87.84%, min;MSMS Calcd.: Calcd.: 809.4; 809.4; MSMS Found: Found: 810.3810.3 [M+H]*.

[M+H].

HPLC

[0641] HPLC

[0641] (Agilent (Agilent HPLC 1200,1200, HPLC Column: Column: Waters Waters X-Bridge X-Bridge C18 (150 mm xmm C18 (150 4.6xmm 4.6x mm 3.5x 3.5 pm); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 1.0 mL/min; 1.0 mL/min; MobileMobile Phase:Phase: from from 95% 95% +[water

[water 10 + 10 mMNH4HCO] mM NH4 HCO 3] and and 5%5% [CH 3CN]

[CHCN] to 0% to 0% [water++10

[water 10 mM mMNH4HCO] NH 4HCOand 3] and 100% 100% [CH 3CN]

[CHCN] in 10 in 10 min, then min, then under under this this condition condition for for 55 min, min, finally finallychanged changed to to95% [water ++ 10 95% [water 10mM NH 4 HCO 3] mM NH4HCO]

and 5%[CHCN] and 5% [CH 3CN] in 0.1 in 0.1 min min and under and under this this condition condition for for 5 min). 5 min). Purity Purity is 84.56%, is 84.56%, = 10.161 Rt =Rt10.161

min. min.

[06421

[0642] ¹H NMR (400 1H NMR MHz, (400 DMSO-d) MHz, 1.12 (6H, DMSO-d) s), (6H, 6 1.12 1.21 s), (6H, s),(6H, 1.21 s), 1.43-1.54 1.43-1.54 (4H, (4H, m), 1.74- in), 1.74 1.78 1.78 (2H, (2H, in), m), 1.88-1.91 (1H, m), 1.88-1.91 (1H, 2.30-2.44 (8H, in), 2.30-2.44 (8H, m), 2.90-2.97 (3H, in), 2.90-2.97 (3H,m), 3.42-3.59(7H, in),3.42-3.59 (7H,m), 4.03 in),4.03-

4.07 (3H, 4.07 (3H,m), in), 4.30 4.30 (1H, (1H, s), s), 6.86-6.91 6.86-6.91 (3H,6.99-7.02 (3H, m), in), 6.99-7.02 (1H, in), 7.22 (1H, m), 7.22 (1H, (1H, d, J= d, J = 2.4 Hz), 2.4 7.64 Hz), 7.64 (1H, d, JJ= (1H, d, 8.8Hz), = 8.8 Hz),7.79 7.79 (1H, (1H, d, Jd,= J= 8.8 8.8 Hz), Hz), 7.90-7.97 7.90-7.97 (2H, (2H, m), in), 8.62 8.62 (1H, d, J(1H, = 2.0d,Hz), J=10.90 2.0 Hz), 10.90 (IH, s). (1H, s).

[0643] Chemical

[0643] Formula: C ChemicalFormula: 4 4 H 52 ClN CHCINO, 7 0, Molecular Molecular Weight: Weight: 810.38. 810.38.

[0644] Total

[0644] Total H count HNMR fromHNMR H countfrom data: 52.52. data:

[0645] Exemplary

[0645] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 47 Compound 47 H ZI H OO N O O O N N N ' N OOt O O N N NCH N ZI H N yJ:N N C1 a o",N CI CI 0 O' 0

[0646] N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-

[0646] N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5 ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1,2,3,4-tetrahydroisoquinolin-6 ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1,2,3,4-tetrahydroisoquinolin-6-

yl)oxy)pentyl)piperazin-1-yl)nicotinamide yl)oxy)pentyl)piperazin-1-yl)nicotinamide

[0647] Synthetic

[0647] Scheme Synthetic Scheme

261

0 0 0 0

0 OH 1) BBrDCM, 1) BBr, rt,o/n DCM, rt, o/n H 1).THF, LDA,-78 rt., 1). THF, LDA, -78 °C,~ C,~rtn o/n OH SOCI 2 SOCl O OH OH O MeOH,rt, MeOH, rt, 1h 1h 0 2) NaOH, 2) MeOH,refluxed, NaOH, MeOH, refluxed, 5h5h HO 2). LiOH.H 2). LiOH.HO,0DMSO, DMSO,120120 °C,°C, 2h 2 h 0 OH OH 66% 0 O 2 66% 61% two 61% twosteps steps H 73%two 73% twosteps steps 0 OH O

<"NH NH N-,, H N N IZ

N TsONCI TsO CI C CI N

CO DMSO, 1).KKCO, 1). 2 3 70 70°C, , DMSO, °C, o/n O/n O SO sCl 2 SOCI N, KI, DIEA 2) LiOH.H 2) 2 0,MeOH, LiOH.HO, MeOH,rt,rt,o/n o/n CI MeOH,rt, MeOH, rt, 2h 2h C CI KI,DIEA 44%two 44% twosteps steps 0 O OH OH 62% 62% 0 DMSO, 7070°C, 0'DIMSO, °C,o/n o/n 41% 41%

0 H2N O OH OH NH H 2N NH HCI HCI

N- H N O"OHATU, HATU, DIEA N IZ LiOH.HO N, N NN 0 1---.-2 N-, H0 I N , 0 0 MeOH,rt, MeOH, rth 3h NN IZ N N- - DMF, rt, 30 min Frt,3 Emin cIao CI 0 N1 NN 94% 94% 81% CI

HCHO 0 H ON IZ

0 HN HN N rN N O N 2.5NNaOH 2.5 N NaOH N N H NN - NN ~ 0 0 DMSOt,5min 5NN DMSO, rt, 5 min NC, N H Nt IZ

N N 42%

CI ~ N42% CI C

[0648]

[0648] Step 1: Step Synthesis of 1: Synthesis 2-(carboxymethyl)-4-methoxybenzoic acidacid of 2-(carboxymethyl)-4-methoxybenzoic

O 0 o 0 0 0 OH o OH O OH 1). THF, LDA, 1). THF, -78°C,~ LDA,-78 rt., o/n °C,-rt., o/n OH - 0 O O 2). 2). LiOH.H 2 0, DMSO, LiOH.HO, DMSO,120120 °C,2 2h h °C, 0 OH 73% two two steps steps O OH 73%

[0649]

[0649] To To aa solution solution of of of of 4-methoxy-2-methylbenzoic acid 4-methoxy-2-methylbenzoic acid (5.0g,g,30.1 (5.0 30.1mmol) mmol)in in drydry

tetrahydrofuran (50 (50 mL) mL)was wasadded added lithium lithium diisopropylamide diisopropylamide in tetrahydrofuran in tetrahydrofuran (1.0 (1.0

mol/L)(66.3 mL,66.3 mol/L)(66.3 mL, 66.3mmol) mmol)at at -78-78 °C °C under under nitrogen nitrogen gas. gas. TheThe mixture mixture was was leftleft to to stirfor stir for 11 hour hour at that at thattemperature temperature and and then then dimethyl carbonate (2.98 dimethyl carbonate (2.98 g, g, 33.1 33.1 mmol) wasadded. mmol) was added.The The reaction reaction

mixture was mixture wasleft left to to stir stirovernight. overnight.Water Water(200 (200 mL) and ethyl mL) and ethyl acetate acetate (100 (100 mL) wasadded. mL) was added.The The aqueous layer was aqueous layer wasseparated, separated, extracted extracted with with ethyl ethyl acetate acetate (50 (50 mL 2) and mL xx 2) and neutralized neutralized with with hydrochloric acid (1N) hydrochloric acid (1 N)until until pH pH<4. < 4.The The mixture mixture waswas extracted extracted with with ethyl ethyl acetate acetate (100mL mL (100 x 2). x 2).

The combined The combinedorganic organiclayers layerswere were washed washed withwith saturated saturated brine brine (50.0 (50.0 mL mL x 2), x 2), dried dried over over

anhydrous sodium anhydrous sodium sulfate,filtered, sulfate, filtered, and and concentrated in in vacuo. vacuo. The residue was The residue was dissolved dissolvedin in dimethyl sulfoxide (40 dimethyl sulfoxide (40 mL) mL)and andlithium lithiumhydroxide hydroxide hydrate hydrate (5.06 (5.06 g, g,120.4 120.4mmol) mmol) was was added. added. The The

262

mixture was mixture wasstirred stirred at at 120 120 °C for 22 hour, °C for hour, cooled downtotoroom cooled down roomtemperature temperature andand poured poured into into ice ice-

water (200 mL). water (200 mL).Hydrochloric Hydrochloricacid acid(1N) (1 N) waswas added added until until pH pH <4. < 4. mixture The The mixture was extracted was extracted

with ethyl with ethyl acetate acetate (100 (100 mL 2). The mL Xx 2). combinedorganic The combined organiclayers layerswere werewashed washed with with saturated saturated brine brine

(50.0 mL 2), dried mL Xx2), dried over over anhydrous anhydroussodium sodium sulfate, filtered, sulfate, filtered, and and concentrated in vacuo concentrated in to give vacuo to

2-(carboxymethyl)-4-methoxybenzoic 2-(carboxymethyl)-4-methoxybenzoic acidacid (4.6(4.6 g, 73% g, 73% two two steps) steps) as aasyellow a yellow solid. solid.

[0650] LC-MS

[0650] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); Column Temperature: 40Flow °C; Rate: Flow Rate: 1.5 mL/min; Mobile Mobile Phase: 95%from 95% 2023248067

mm*3.5 µm); Column Temperature: 40 °C; 1.5 mL/min; Phase: from

[water ++ 0.1%

[water 0.1% TFA] TFA] and and 5% [CH 3 CN 5% [CHCN + 0.1% + 0.1% TFA] TFA] to to 0%0% [water+ +0.1%

[water 0.1%TFA] TFA] andand 100% 100%

[CH 3 CN

[CHCN + 0.1% + 0.1% TFA] TFA] in 0.5inmin, 0.5 min, then then underunder this this condition condition for min, for 1.5 1.5 min, finally finally changed changed to 95% to 95%

[water +

[water 0.1%TFA] + 0.1% TFA]andand 5% 5% [CH

[CHCN CN +TFA] + 30.1% 0.1% in TFA] in and 0.1 min 0.1 min underand under this this condition condition for 0.5 for 0.5 min). Purity min). Purity is is94.6%, 94.6%, Rt = 0.774 Rt = min; MS 0.774 min; MSCalcd.: Calcd.:210.1; 210.1;MSMS Found: Found: 233.1 233.1 [M+23]*.

[M+23].

[0651] Step Step

[0651] 2: Synthesis 2: Synthesis of methyl of methyl 4-methoxy-2-(2-methoxy-2-oxoethyl)benzoate 4-methoxy-2-(2-methoxy-2-oxoethyl)benzoate

0 O O SOH 'OHSOC1 OH O SOCl 0 -1 o MeOH,rt,rt,1h1h MeOH, o O O OH 66% O OH 66% O

[0652]

[0652] To To aa solution solution of of (2-(carboxymethyl)-4-methoxybenzoic (2-(carboxymethyl)-4-methoxybenzoic acidacid (1.2 (1.2 g, g, 5.75.7mmol) mmol) in in

methanol(10.0 methanol (10.0 mL) mL)was was added added thionyl thionyl chloride chloride (1.7g,g,14.3 (1.7 14.3mmol) mmol) dropwise. dropwise. The The mixture mixture was was refluxed for refluxed for 22 hour. hour. The The mixture was cooled mixture was cooleddown downto toroom room temperature temperature and and thenthen the the solvent solvent waswas

removedininvacuo removed vacuototogive givecrude crudeproduct productwhich whichwaswas purified purified by by column column chromatography chromatography on silica on silica

gel (ethyl (ethyl acetate/ acetate/petroleum petroleum ether ether==1:1:1)1) to to give 4-methoxy-2-(2-methoxy-2 give 4-methoxy-2-(2-methoxy-2-

oxoethyl)benzoate(900 mg, oxoethyl)benzoate(900 mg, 66%) 66%) aswhite as a a white solid. solid.

[0653] Step Step

[0653] 3: Synthesis 3: Synthesis of 2-(carboxymethyl)-4-hydroxybenzoic of 2-(carboxymethyl)-4-hydroxybenzoic acid acid o o 0 o 0 1) BBr, 1) BBr3DCM, , DCM, rt, rt, o/no/n OH OH o 2) NaOH, 2) MeOH,refluxed, NaOH, MeOH, refluxed, 55 hh HO Ho O O two steps 61% two 61% steps 0 OH O O OH

[0654] To a To

[0654] a solution solution of 4-methoxy-2-(2-methoxy-2-oxoethyl)benzoate of 4-methoxy-2-(2-methoxy-2-oxoethyl)benzoate (0.9 g, (0.9 g, 3.78 3.78inmmol) mmol) in dichloromethane (30mL) dichloromethane (30 mL)waswas added added boron boron tribromide tribromide (4.7 (4.7 g, 18.9 g, 18.9 mmol) mmol) dropwise dropwise under under ice- ice

waer bath. waer bath. The The resulting resulting mixture mixture was wasallowed allowedtotowarm warmto to room room temperature temperature and and stirred stirred overnight. overnight.

Water (100 mL) Water (100 mL)waswas added. added. TheThe organic organic layer layer waswas separated, separated, washed washed with with brinebrine (50xmL (50 mL 2), x 2),

263 dried dried over over anhydrous sodiumsulfate, sulfate, filtered, filtered, and and concentrated concentrated in in vacuo to give give aa mixture. mixture. The 2023248067 10 Oct 2023 anhydrous sodium vacuo to The mixture was mixture wasdissolved dissolvedininmethanol methanol(30 (30mL) mL)andand sodium sodium hydroxide hydroxide (0.76(0.76 g, 18.9 g, 18.9 mmol) mmol) in water in water

(4.0 (4.0 mL) wasadded. mL) was added.The Themixture mixture was was refluxed refluxed forfor 5 hour.TheThe 5 hour. solvent solvent waswas removed. removed. The The residue residue

was dissolved was dissolved in in water water (30 (30 mL). mL).Hydrochloric Hydrochloricacid acid(1N) (1 N) waswas added added until until pH pH <4. <The 4. mixture The mixture was extracted with was extracted with ethyl ethyl acetate acetate (50 (50 mL 2). The mL Xx 2). combinedorganic The combined organiclayers layerswere werewashed washed with with

saturated brine(20.0 saturated brine (20.0mL mL 2), dried x dried X 2), over over anhydrous anhydrous sodiumfiltered, sodium sulfate, sulfate, and filtered, and concentrated concentrated in in vacuo to give vacuo to give 2-(carboxymethyl)-4-hydroxybenzoic 2-(carboxymethyl)-4-hydroxybenzoic acidacid (0.45 (0.45 g, 61% g, 61% two two steps) steps) as aasyellow a yellow solid. solid.

[0655] LC-MS

[0655] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 1.5 mL/min; 1.5 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water ++0.1%

[water 0.1% TFA] TFA] and and 5% [CH 3 CN 5% [CHCN + 0.1% + 0.1% TFA] TFA] to to 0%0% [water+ +0.1%

[water 0.1%TFA] TFA] andand 100% 100%

[CH 3 CN

[CHCN + 0.1% + 0.1% TFA] TFA] in 0.5inmin, 0.5 min, then then underunder this this condition condition for min, for 1.5 1.5 min, finally finally changed changed to 95% to 95%

[water ++ 0.1%

[water 0.1%TFA] TFA]andand 5% 5% [CH

[CHCN CN +TFA] + 30.1% 0.1% in TFA] in and 0.1 min 0.1 min underand under this this condition condition for 0.5 for 0.5 min). Purity min). Purity isis95.2%, 95.2%, Rt = 0.570 Rt = min; MS 0.570 min; MSCalcd.: Calcd.:196.0; 196.0;MSMS Found: Found: 197.2 197.2 [M+H]*.

[M+H].

[0656]

[0656] Step 4: Step 4: Synthesis of 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)acetic Synthesis of acid 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)acetic acid

O TsO TsO C1 CI O - 2CO 3, DMSO, 70 °C, o/n OH 1). K 0 OH 1). KCO, DMSO, 70 °C, o/n HOJ 2) LiOH.H CI Ho 2) 2 0,MeOH, LiOH.HO, MeOH, rt, o/n rt, o/n o 0 OH 44%two 44% two steps steps O O OH OH O OH

[0657] The

[0657] The mixture mixture of 2-(carboxymethyl)-4-hydroxybenzoic of 2-(carboxymethyl)-4-hydroxybenzoic acid (120acid mg, (120 0.61 mg, 0.61 mmol), mmol),

potassiumcarbonate potassium carbonate(253 (253mg, mg,1.83 1.83mmol) mmol) and and 5-chloropentyl 5-chloropentyl 4-methylbenzenesulfonate 4-methylbenzenesulfonate (506 (506 mg, 1.83 mg, 1.83 mmol) mmol)inindimethyl dimethylsulfoxide sulfoxide(5(5mL) mL) waswas stirred stirred atat7070°C°C overnight.The overnight. Theresulting resulting mixture was mixture wasallowed allowedtotocooled cooleddown downto to room room temperature temperature and and stirred stirred overnight. overnight. Water Water (20 (20 mL) mL)

and ethyl acetate and ethyl acetate (20 (20 mL) was added. mL) was added.The Theorganic organiclayer layerwas wasseparated, separated,washed washed with with brine brine (50(50

mL mL X x 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a

mixture. The mixture. The mixture mixturewas wasdissolved dissolvedininmethanol methanol(30(30mL)mL) and and lithium lithium hydroxide hydroxide hydrate hydrate (128 (128 mg, mg, 3.05 3.05 mmol) wasadded. mmol) was added.TheThe mixture mixture waswas stirred stirred at at room room temperature temperature overnight. overnight. The The solvent solvent was was

removed.The removed. Theresidue residuewas wasdissolved dissolvedin inwater water(30(30mL). mL).Hydrochloric Hydrochloric acidacid (1 was (1N) N) added was added until until

pH<4. pH < 4.The Themixture mixture waswas extracted extracted with with ethyl ethyl acetate(20(20mLmL acetate x 2). X 2). TheThe combined combined organic organic layers layers

were washedwith were washed withsaturated saturatedbrine brine(10 (10 mLmL x 2),dried X 2), driedover overanhydrous anhydrous sodium sodium sulfate,filtered, sulfate, filtered, and and

264

Oct 2023 concentrated in vacuo concentrated in to give vacuoto 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)acetic acidacid give 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)acetic (85 mg, (85 two 44%two mg, 44% steps)asasyellow steps) oil. yellowoil.

[0658] LC-MS

[0658] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90% 2023248067 10

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 10% 10% [CH 3 CN]to

[CHCN] to 5% [water + +1010 5% [water mMmMNH 4HCO 3 ] and NH4HCO] and 95% 95%

[CH 3 CN]

[CHCN] in 0.5 in 0.5 min, min, then then under under thiscondition this condition for1.5 for 1.5min, min,finally finally changed changedtoto90% 90% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 310% 10% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this for condition condition for 0.5 0.5 min.). min.).is Purity Purity is 69.9%, Rt =0.829 69.9%, Rt =0.829min; min;MSMS Calcd.: Calcd.: 314.1; 314.1; MS MS Found: Found: 315.1315.1 [M+H]*.

[M+H].

[0659] Step Step

[0659] 5: Synthesis 5: Synthesis of methyl of methyl 4-(5-chloropentyloxy)-2-(2-methoxy-2 4-(5-chloropentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoate oxoethyl)benzoate

0 O O O SOCI 2 O SOCl CI O MeOH,rt,rt,2 2hh MeOH, CI

0 O OH OH 62% 62% 0 0 O

[0660] To a To

[0660] a solution solution of 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)acetic of 2-(5-(5-chloropentyloxy)-2-(methoxycarbonyl)phenyl)aceti acid (85acid (85 mg, 0.27 mg, 0.27 mmol) mmol)ininmethanol methanol(2 (2 mL) mL) waswas added added thionyl thionyl chloride chloride (48.3 (48.3 mg, mg, 0.410.41 mmol) mmol) dropwise. dropwise.

The mixturewas The mixture wasrefluxed refluxedfor for22hour. hour. The Themixture mixturewas was cooled cooled down down to room to room temperature temperature and and

then then the the solvent solvent was removedininvacuo was removed vacuototogive givecrude crudeproduct productwhich which was was purified purified by by prep-TLC prep-TLC

(ethyl (ethyl acetate/ acetate/petroleum petroleum ether ether== 1:1:1)1)toto give methyl give methyl4-(5-chloropentyloxy)-2-(2-methoxy-2 4-(5-chloropentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoate (55mg, oxoethyl)benzoate (55 mg,62%) 62%)as as yellow yellow oil. oil.

[0661] LC-MS

[0661] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 10% 10% [CH 3 CN]to

[CHCN] to 5% [water + +1010 5% [water mMmMNH 4HCO 3 ] and NH4HCO] and 95% 95%

[CH 3 CN]

[CHCN] in 0.5 in 0.5 min, min, then then under under thiscondition this condition for1.5 for 1.5min, min,finally finally changed changedtoto90% 90% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 310% 10% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this for condition condition for 0.5 0.5 min.). min.).is Purity Purity is 72.9%, Rt =1.208 72.9%, Rt min;MSMS =1.208min; Calcd.: Calcd.: 328.1; 328.1; MS MS Found: Found: 329.2329.2 [M+H]*.

[M+H].

[0662] Step Step

[0662] 6: Synthesis 6: Synthesis of methyl of methyl 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 4-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoate oxoethyl)benzoate

265

2023248067 10 2023

r"NH 0 NH N N o NC N O H HN N IN n zN N Oct o "' < CI CI 0" N- N 0 o KI, DIEA CI KI, DIEA NN

O O / DMSO, DMSO,7070°C, °C, o/n o/n H HN IN N 41% N 41% N Na o O" CI CI

[0663] The mixture

[0663] The mixture of methyl of methyl 4-(5-chloropentyloxy)-2-(2-methoxy-2-oxoethyl)benzoate 4-(5-chloropentyloxy)-2-(2-methoxy-2-oxoethyl)benzoate (55 (55 mg, 0.17 mg, 0.17 mmol), ethyldiisopropylamine mmol),ethyldiisopropylamine (65.8 (65.8 mg,mg, 0.51 0.51 mmol), mmol), potassium potassium iodide (28.2(28.2 iodide mg, mg, 0.17 0.17 mmol)and mmol) and-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(piperazin- N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(piperazin 1-yl)nicotinamide (78.5 (78.5 mg, mg, 0.17 0.17 mmol) mmol)inindimethyl dimethylsulfoxide sulfoxide(2(2mL) mL) waswas stirred stirred atat7070°C°C

overnight. overnight. The resulting mixture The resulting was allowed mixture was allowedtotocooled cooleddown downto to room room temperature temperature and and stirred stirred

overnight. overnight. Water (20 mL) Water (20 mL)and andethyl ethylacetate acetate (20 (20mL) mL)was was added. added. TheThe organic organic layer layer waswas separated, separated,

washed withbrine washed with brine(50 (50mLmL x 2),dried x 2), driedover overanhydrous anhydrous sodium sodium sulfate,filtered, sulfate, filtered, and andconcentrated concentrated in in vacuo to give vacuo to give the the crude crude product product which waspurified which was purified by bycolumn columnand and flashchromatography flash chromatography (ethyl acetate/ petroleum (ethyl acetate/ petroleum ether ether 1: to1)give = 1:= 1) to give methyl methyl 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4 4-(5-(4-(5-(1r,3r)-3-(3-chloro-4-

cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy) cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcaibamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy).-

2-(2-methoxy-2-oxoethyl)benzoate 2-(2-methoxy-2-oxoethyl)benzoate (53(53 mg,mg, 41%)41%) as a as a white white solid. solid.

[0664] Step Step

[0664] 7: Synthesis 7: Synthesis of 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of 4-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoic acid oxoethyl)benzoic acid

0/ HO HO o o 0 0 00 0 o O o IN N NN IN CD) N N LiOH.H 20 LiOH.HO N MeOH, rt, 3 h MeOH, rt, 3 h N HNNP HN 81% 81% HN HN o O NIN/ NE N CI CI CI CI

266

[0665] The mixture

[0665] The mixture of methyl of methyl 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 4-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoate (53mg, oxoethyl)benzoate (53 mg,0.07 0.07mmol) mmol)waswas dissolved dissolved in methanol in methanol (2 mL) (2 mL) and lithium and lithium hydroxide hydroxide

hydrate (14.7 mg, hydrate (14.7 0.35 mmol) mg, 0.35 mmol)was was added. added. TheThe mixture mixture was was stirred stirred at at room room temperature temperature for for 3 3

hour. The hour. solvent was The solvent was removed. removed.The The residue residue was was dissolved dissolved in in water water (15(15 mL). mL). Hydrochloric Hydrochloric acidacid

(1 (1 N) N) was addeduntil was added until pH pH<4. < 4.The The mixture mixture waswas extracted extracted with with ethyl ethyl acetate(15(15mLmL acetate x 2). x 2). TheThe

combined organiclayers combined organic layerswere werewashed washed with with saturated saturated brine brine (10(10 mL mL x 2), X 2), dried dried over over anhydrous anhydrous

sodium sulfate,filtered, sodium sulfate, filtered,andand concentrated concentrated in vacuo in vacuo to giveto4-(5-(4-(5-(1r,3r)-3-(3-chloro-4 give 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4 cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy) cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcatbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-

2-(2-methoxy-2-oxoethyl)benzoic 2-(2-methoxy-2-oxoethyl)benzoic acid acid (42(42 mg,mg, 81%) 81%) as a as a white white solid. solid.

[0666] LC-MS

[0666] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 10% 10% [CH 3 CN]to

[CHCN] to 5% [water + +1010 5% [water mMmMNH and 95% 4HCO 3 ] and NH4HCO] 95%

[CH 3 CN]

[CHCN] in 0.5 in 0.5 min, min, then then under under thiscondition this condition for1.5 for 1.5min, min,finally finally changed changedtoto90% 90% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 310% 10% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this for condition condition for 0.5 0.5 min.). min.).is Purity Purity is 75.4%, Rt =1.041 75.4%, Rt =1.041min; min;MSMS Calcd.: Calcd.: 745.3; 745.3; MS MS Found: Found: 746.2746.2 [M+H]*.

[M+H].

[0667] Step Step

[0667] 8: Synthesis 8: Synthesis of methyl of methyl 2-(5-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy) 2-(5-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)

2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2,6 2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2,6-

dioxopiperidin-3-ylcarbamoyl)phenyl)acetate dioxopiperidin-3-ylcarbamoyl)phenyl)acetate 0H IZ

H2N OH OH H2N NH N HCI HCI HC HN O N N O HATU, DIEA N - 0 N HATU, DIEA N N N, OI DMF, rt, 30 min NN IZ DMF, rt, 30 min

N N~~ y -~ 9%N 94% N IZ

N CI 0C1 O NC O O- N CI

[0668] A solution

[0668] A solution of 4-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of 4-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2-methoxy-2-

oxoethyl)benzoic acid(42 oxoethyl)benzoic acid (42 mg, mg,0.056 0.056mmol), mmol), HATU HATU (25.5(25.5 mg, 0.067 mg, 0.067 mmol) mmol) and and ethyldiisopropylamine (29.7mg, ethyldiisopropylamine (29.7 mg,0.23 0.23mmol) mmol)in in N, N, N-dimethylformamide N-dimethylformamide (2was (2 mL) mL)stirred was stirred for for

30 min, and 30 min, and then then 3-aminopiperidine-2,6-dione 3-aminopiperidine-2,6-dionehydrochloride hydrochloride (9.2 (9.2 mg, mg, 0.056 0.056 mmol) mmol) was added. was added.

The mixture The mixturewas wasstirred stirred at at room temperatureovernight room temperature overnightand andwater water (10mL)mL) (10 waswas added. added. The The mixture was mixture wasextracted extracted by byethyl ethyl acetate acetate (20 (20 mL 3). The mL X x3). Thecombined combined organic organic layerswere layers were washed washed

with brine(10 with brine (10mLmL x 3), X 3), dried dried over over anhydrous anhydrous sodium filtered, sodium sulfate, sulfate, filtered, and concentrated and concentrated in vacuo. in vacuo.

267

Oct 2023 The residue was The residue waspurified by prep-TLC purified by prep-TLC (dichloromethane/ (dichloromethane/ methanol=10:1) methanol=10:1) to give to give methyl methyl 2-(5-2-(5

(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin (5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutylcarbamoyl)pyridin-

2-yl)piperazin-1-yl)pentyloxy)-2-(2,6-dioxopiperidin-3-ylcarbamoyl)phenyl)acetate 2-yl)piperazin-1-yl)pentyloxy)-2-(2,6-dioxopiperidin-3-ylcarbamoyl)phenyl)acetate (45(45 mg,mg,

94%) 94%) asas a a white white solid. solid. 2023248067 10

[0669] LC-MS

[0669] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (30 (30 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 10% 10% [CH 3 CN]to

[CHCN] to 5% [water + +1010 5% [water mMmMNH 4HCO 3 ] and NH4HCO] and 95% 95%

[CH 3 CN]

[CHCN] in 0.5 in 0.5 min, min, then then under under thiscondition this condition for1.5 for 1.5min, min,finally finally changed changedtoto90% 90% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 310% 10% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this for condition condition for 0.5 0.5 min.). min.).isPurity Purity is 77.7%, Rt =1.213 77.7%, Rt =1.213min; min;MSMS Calcd.: Calcd.: 855.4; 855.4; MS MS Found: Found: 856.3856.3 [M+H]*.

[M+H].

[0670] Step Step

[0670] 9: Synthesis 9: Synthesis of N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1,2,3,4 tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1,2,3,4-

tetrahydroisoquinolin-6-yloxy)pentyl)piperazin-1-yl)nicotinamide tetrahydroisoquinolin-6-yloxy)pentyl)piperazin-1-yl)nicotinamide

N N N N CI CI CI CI

0 0. O,, O,,

NH NH O NH NH 0 O 2.5 2.5 NN NaOH NaOH 0 o NN N DMSO, rt, 5 min DMSO, rt, 5 min N N N N K.NN 42% N 42% N

o 0 H HN o NH0 o NH o 0 oo o0 O1 o o O O N O 110 NH NH 0 _ O

[0671] A solution

[0671] A solution of methyl of methyl 2-(5-(5-(4-(5-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 2-(5-(5-(4-(5-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2,6-dioxopiperidin-3 tetramethylcyclobutylcarbamoyl)pyridin-2-yl)piperazin-1-yl)pentyloxy)-2-(2,6-dioxopiperidin-3-

ylcarbamoyl)phenyl)acetate(45 ylcarbamoyl)phenyl)acetate (45mg, mg, 0.053 0.053 mmol) mmol) in dimethyl in dimethyl sulfoxide sulfoxide (2 mL) (2 mL) was added was added

sodium hydroxideininwater sodium hydroxide water(2.5 (2.5moL/L, moL/L, 2 drops).The 2 drops). The mixture mixture waswas stirred stirred at at room room temperature temperature

for for 55 min. min. Water (20 mL) Water (20 mL)and andethyl ethylacetate acetate (20 (20 mL) mL)was wasadded. added. The The organic organic layer layer waswas separated, separated,

washed withbrine washed with brine(10 (10 mLmL x 2),dried X 2), driedover overanhydrous anhydrous sodium sodium sulfate,filtered, sulfate, filtered, and andconcentrated concentrated

268 in vacuo to give the the crude product which crude product waspurified byprep-HPLC purified by prep-HPLCto to give N-((r,3r)-3-(3 10 Oct 2023 vacuo to which was give N-((1r,3r)-3-(3- chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl) chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-

1,3-dioxo-1,2,3,4-tetrahydroisoquinolin-6-yloxy)pentyl)piperazin-1-yl)nicotinamide 1,3-dioxo-1,2,3,4-tetrahydroisoquinolin-6-yloxy)pentyl)piperazin-l-yl)nicotinamide (18.5mg, (18.5 mg, 42%)asasaa white 42%) white solid. solid.

[0672] LC-MS

[0672] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[(total 10mM 2023248067

[(total 10mM AcONH AcONH4)4)water/CHCN=900/100 water/CH3CN=900/100 (v/v)] (v/v)] andand10%10% [(total 10mM

[(total AcONH 4 10mM AcONH4)

) water/CH3CN=100/900 (v/v)]toto 10% water/CHCN=100/900 (v/v)] 10%[(total

[(total 10mM AcONH water 10mM AcONH4) 4) water /CH 3 CN=900/100 /CHCN=900/100 (v/v)] (v/v)]

and 90%[(total and 90% [(total 10mM 10mM AcONH AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] (v/v)] in 1.6 in then min, 1.6 min, underthen under this this

condition for for 2.4 2.4 min, min, finally finallychanged changed to to 90% [(total10mM 90% [(total AcONH 10mM AcONH4) 4) water/CH3CN=900/100 water/CHCN=900/100

(v/v)] (v/v)] and and 10% [(total10mM 10% [(total AcONH 10mM AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] in (v/v)] 0.1 minin and 0.1 under min and under this this

condition for 0.7 condition for 0.7 min). min). Purity Purity isis100.0%, 100.0%, Rt Rt == 2.988 2.988 min; min; MS Calcd.:823.4;MS MS Calcd.:823.4; MS Found:824.3 Found:824.3

[M+H]*.

[M+H].

[0673] HPLC

[0673] HPLC (Agilent (Agilent HPLCHPLC 1200;1200; Column: Column: L-column2 L-column2 ODS mm*4.6 ODS (150 (150 mm*4.6 mm*5.0 mm*5.0 µm); m); Column Temperature: Column Temperature: 40 40 °C;°C; Flow Flow Rate: Rate: 1.0 1.0 mL/min; mL/min; Mobile Mobile Phase:Phase: from from 95% 95% +[water

[water 0.1% + 0.1%

TFA] and TFA] and 5% 5%[CHCN

[CH 3 CN + 0.1% + 0.1% TFA] TFA] to 0% to 0% [water

[water + 0.1% + 0.1% TFA] TFA] andand 100% 100% [CH

[CHCN CN + 0.1% + 3 0.1%

TFA]inin1010min, TFA] min,then thenunder underthis this condition condition for for 55 min, finally changed min, finally to 95% changed to [water++0.1% 95% [water 0.1%TFA] TFA] and 5%[CHCN and 5% [CH 3+CN + 0.1% 0.1% TFA] TFA] in 0.1 in 0.1and min min and this under undercondition this condition for 5 min). for 5 min). PurityPurity is 95.2 %, is 95.2%,

Rt = 8.168 Rt 8.168 min. min.

[0674]

[0674] ¹H NMR MHz,MHz, (400 (400 'H NMR DMSO-d) 1.12 6)(6H, DMSO-d 6 1.12 (6H, s), 1.21 1.21 (6H, s), (6H, s), 1.37-1.58 s), 1.37-1.58 (4H,m), (4H, 1.73 m),1.73- 1.81 1.81 (2H, (2H, m), m), 1.86-1.91 (1H, m), 1.86-1.91 (1H, m), 2.30-2.37 2.30-2.37 (2H, (2H, m), m), 2.40-2.46 2.40-2.46 (2H, (2H,m), m),2.82-2.91(1H, 2.82-2.91(1H,m), m),3.30- 3.30 3.35 (4H, m), m), 3.55-3.65 3.55-3.65 (4H, (4H, m), m), 4.03-4.30 4.03-4.30 (6H, (6H, m), m),5.54-5.63 (1H, m), 5.54-5.63 (1H, m),6.87 (1H,d,d,JJ= 6.87(1H, 9.6 Hz), = 9.6 Hz), 6.96-7.07 (3H,m),m), 6.96-7.07 (3H, 7.21 7.21 (1H,(1H, d, J d, J=Hz), = 2.4 2.4 7.63 Hz), (1H, 7.63d,(1H, d, J= J = 9.6 Hz),9.6 Hz), 7.90-8.04 7.90-8.04 (3H, m), 8.62 (3H, m), 8.62

(1H, d, JJ= (1H, d, 2.4Hz), = 2.4 Hz),10.93 10.93 (1H,(1H, s). s).

[0675] Chemical

[0675] Formula: C ChemicalFormula: CHCINO, 70 7 , Molecular 4 4 H5 0 ClNMolecular Weight: Weight: 824.36. 824.36. Total

[0676] Total

[0676] H count H count from HNMR fromHNMR data: 50.50. data:

[0677] Exemplary

[0677] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 48 Compound 48

269

Oct 2023 00 0 N NH N O N N NN N N ZI H N N 2023248067 10

CI "O o

N-((1 r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-((2-(2,6 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,24,4-tetramethylcyclobutyl)-6-(4-(5-((2-(2,6-

dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-11,2,41triazolo14,3-alpyridin-7 dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-|1,2,4]triazolo[4,3-a|pyridin-7-

yl)oxy)pentyl)piperazin-1-yl)nicotinamide yl)oxy)pentyl)piperazin-1-yl)nicotinamide

106781

[0678] Synthetic Scheme Synthetic Scheme Br N N NCl Z'N CI N 2H 4.H 20,EtOH EtOH NH N NH.HO, IZ NH HO HON i CI 60% NaH, DMF 60% NaH, DMF M,2C8h MW, 120°C, 8 h H 50 °C, 50 *C, o/n o/f 83% 83% 78% 78%

0 0 O 0 0 O NH o o CDI, N Br N NH CDI,CHCHCN 3 CN NH ___NH____________ N O 80 'C, 22hh 80 °C, N o - o o N N K2 C03 , CH KCO, 3CN CHCN .....o--- lo,' z N 20% 80 80 °C, /n *C, o/n 20% 39% 39%

NH NH H -rN N NN N HN

N 0N N o o N- NH CI o 3 , rt,0/ CHCIrt, TMSI,CHCl, TMSI, o/n N N 7 o 0______________ Ó 0 79% 1 0DIEA, o N DIEA, CH 3CN, CHCN, 8080°C, /n C, o/n 79% 34% 34%

0 0 o o N NH N -0 N o N o N N~ N N N ZI

N N

CIa CI 0N- 0 O

106791 Step Step

[0679] 1: Synthesis 1: Synthesis of 4-(5-(benzyloxy)pentyloxy)-2-chloropyridine of 4-(5-(benzyloxy)pentyloxy)-2-chloropyridine

270

~Br 2023248067 10 Oct 2023

O Br N N -N N O' O CI CI O HO CI CI 60% DMF NaH, DMF 60% NaH, Ho 50 o/n °C, o/n 50 °C, 78% 78%

[0680] To a To

[0680] a solution solution of 2-chloropyridin-4-ol (1.3(1.3 of 2-chloropyridin-4-ol g, 10.0 g, 10.0 mmol) mmol) in DMF in DMF (15wasmL) (15 mL) was added added sodium hydride(60% sodium hydride (60% dispersed dispersed in in mineral mineral oil, 482mg, oil,482 mg,12.0 12.0mmol) mmol) at 0at°C, 0 °C, andand thethe mixture mixture waswas

stirred stirred atatroom room temperature temperature for for 30 30 min. min. Then ((5-bromopentyloxy)methyl)benzene Then ((5-bromopentyloxy)methyl)benzene (3.1 (3.1 g, 12.0 g, 12.0

mmol)was mmol) wasadded added to to thereaction the reactionand andthe theresulted resultedmixture mixturewas wasstirred stirred at at 50 50 °C °C overnight. overnight. When When the reaction the reaction was was completed (monitoredbybyTLC), completed (monitored TLC), water water (30(30 mL)mL) was was added. added. The resultant The resultant

mixture was mixture wasextracted extracted by byethyl ethyl acetate acetate (10 (10 mL 3) and mL X x3) andthe the combined combinedorganic organic layerswere layers were washedbybybrine washed brine(20 (20mLmL x 3),dried x 3), driedover overanhydrous anhydrous sodium sodium sulfate,filtered sulfate, filteredand andconcentrated. concentrated. The residue was The residue waspurified purified by by column columnchromatography chromatography on silica on silica (petroleum/ethyl (petroleum/ethyl acetate acetate = 1/4) = 1/4) to to

give give 4-(5-(benzyloxy)pentyloxy)-2-chloropyridine (2.4g,g,78% 4-(5-(benzyloxy)pentyloxy)-2-chloropyridine (2.4 78% yield)asasa abrown yield) brown solid. solid.

[0681]

[0681] ¹H NMR (400 (400 'H NMR MHz, MHz, CDCl)CDC 1.47-1.53 (2 H, m), 1.59-1.64 (2 H, m), 1.71-1.76 (2 H, 3 ) 81.47-1.53 (2 H, m), 1.59-1.64 (2 H, m), 1.71-1.76 (2 H,

m), 3.42 m), 3.42(2(2H,H,t,t,JJ= 6.4Hz), = 6.4 Hz), 3.91 3.91 (2 t, (2 H, H, Jt,=J= 6.4 6.4 Hz),Hz), 4.44 4.44 (2 H, (2 s),H,7.07-7.15 s), 7.07-7.15 (2 H, (2 H, m), m), 7.23- 7.23 7.28 (5 H,H,m), 7.28 (5 m),7.96 7.96 (1 (1H, H, d, Jd,=J= 3.2 3.2 Hz).Hz).

[0682] Step Step

[0682] 2: Synthesis 2: Synthesis of 4-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine of 4-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine

N N f N -~ N 0" - CI NH CI -N 2 H 4 .H 2 0,EtOH EtOH N -. O OIINM OC8 NH.HO, O o IZ NH MW,120°C, MW, 120-C,8 8 hhH H 83% 83%

[0683] To a To

[0683] a microwave microwave glass was glass vial was added vialadded 4-(5-(benzyloxy)pentyloxy)-2-chloropyridine 14-(5-(benzyloxy)pentyloxy)-2-chloropyridine

(2.0 (2.0 g, g,6.5 6.5mmol), mmol), hydrazine monohydrate(10(10mL)mL) hydrazine monohydrate andand EtOHEtOH (10 mL), (10 mL), andmixture and the the mixture was was stirred stirred under under microwave conditionsatat 120 microwave conditions 120°C°Cfor for88 h. h. When Whenititwas wascooled cooledtotoroom room temperature, temperature,

water (20 mL) water (20 mL)was wasadded added to to thereaction. the reaction. The Theresultant resultant mixture mixturewas wasextracted extractedbybyethyl ethylacetate acetate (10 (10 mL 3) and mL X x3) andthe the combined combined organiclayers organic layerswere were washed washed by brine by brine (15 (15 mL XmL 3), dried 3),x dried overover

anhydroussodium anhydrous sodium sulfate,filtered, sulfate, filtered, and and concentrated in in vacuo. vacuo. The residue (1.6 The residue (1.6 g, g, 83% yield) 83% yield)

wasdirectly was directlyused used to to thethe next next stepstep without without further further purification purification as oil. as brown brown oil.

[0684] Step Step

[0684] 3: Synthesis 3: Synthesis of 7-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H) of 7-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-

one one

271 o N N_ N CDI, CHCN NH c O Z 0O NNH2 CDI, CH3CN ,1 NH IZ N NH H 80 °C,2 h o O N H 80 °C, 2h 20% 20%

[0685] To a To

[0685] a solution solution of 5-ethoxy-2-hydrazinylpyridine of 5-ethoxy-2-hydrazinylpyridine (1.65.4 (1.6 g, g, mmol) 5.4 mmol) in acetonitrile in acetonitrile (25 (25 mL)was mL) wasadded added CDI CDI (1.3 (1.3 g, g, 8.28.2mmol), mmol), andand thethe mixture mixture was was stirred stirred at at 8080 °C °C forfor 2 h.When 2 h. When it was it was

cooled to room cooled to temperature, water room temperature, water(20 (20mL) mL)waswas added added to to thethe reaction.The reaction. The resultantmixture resultant mixturewas was extracted extracted by by ethyl ethyl acetate acetate (10 (10mL x 3) and mL X and the the combined organiclayers combined organic layers were werewashed washedby by brine brine

(15 (15 mL 3), dried mL X x3), dried over over anhydrous anhydroussodium sodium sulfate,filtered, sulfate, filtered, and and concentrated in vacuo. concentrated in The vacuo. The

residue was purified by column was purified columnchromatography chromatography on silica on silica (DCM/MeOH (DCM/MeOH 20/1) = 20/1)=to giveto7-(5- give 7-(5 (benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (360 (benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-apyridin-3(2H)-one (360 mg,mg, 20% 20% yield) yield) as aas a white white

solid. solid.

[0686] LC-MS

[0686] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 4.6 x 4.6 C18mm(50x mm mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 96.77%, Rt==1.716 96.77%, Rt 1.716min. min.MSMS Calcd.: Calcd.: 327.16; 327.16; MS MS Found: Found: 328.2328.2 [M+H]*.

[M+H].

[0687] Step Step

[0687] 4: Synthesis 4: Synthesis of 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3 of 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-

a]pyridin-2(3H)-yl)piperidine-2,6-dione a]pyridin-2(3H)-yl)piperidine-2,6-dione

0 o o N 00o NH O N Br N~N NH NH O';::-N-NH NH Brt~ 0 N O ^ O N K 2CO 3 ,CH 3CN O O N O O KCO, CHCN O o 0 ~80*C,ao/n 80 °C, o/n

39% 39%

[0688] The solution

[0688] The solution of 7-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one of 7-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one

(300 mg, mg, 0.9 0.9 mmol), mmol),3-bromopiperidine-2,6-dione 3-bromopiperidine-2,6-dione (438 (438 mg, mg, 2.3 2.3 mmol) mmol) and(253 and KCO K2 C03 mg, (253 1.8 mg, 1.8

mmol)ininacetonitrale mmol) acetonitrale (10 (10 mL) mL)was wasstirred stirred at at 80 80 °C °C overnight. overnight. When Whenititwas wascooled cooledtotoroom room temperature, water (10 temperature, water (10 mL) mL)was wasadded. added.TheThe resultantmixture resultant mixture was was extracted extracted by by ethyl ethyl acetate(10 acetate (10 mL 3) and mL X x3) andthe the combined combined organiclayers organic layerswere were washed washed by brine by brine (10 (10 mL xmL 3), dried 3),x dried overover

anhydrous sodium anhydrous sodium sulfate,filtered sulfate, filtered and and concentrated. The residue concentrated. The residue was waspurified purified by by Prep-TLC Prep-TLC (DCM/MeOH = 20/1) (DCM/MeOH = 20/1) to give to give 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-

a]pyridin-2(3H)-yl)piperidine-2,6-dione (157 a]pyridin-2(3H)-yl)piperidine-2,6-dione (157 mg, mg,39% 39% yield) yield) as as a awhite whitesolid. solid.

272

X-Bridge 4.6 x 4.6 C18mm(50X mm 2023248067 10 2023

[0689] LC-MS

[0689] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge C18 (50

mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

Oct [water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed 95% changedtoto95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 99.45%,RtRt==1.836 99.45%, 1.836min. min.MSMS Calcd.: Calcd.: 438.19; 438.19; MS MS Found: Found: 439.3439.3 [M+H]*.

[M+H].

[0690] Step Step

[0690] 5: Synthesis 5: Synthesis of 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin of 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-

2(3H)-yl)piperidine-2,6-dione 2(3H)-yl)piperidine-2,6-dione

o NH0 0 0 NH 0 o o N N N NH N o N o'o TMSI, CHCl, rt, o/n TMSI, CHCl 3 , rt, o/n N o N o N 79% 79%

[0691] To a To

[0691] a solution solution of 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin of 3-(7-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-

2(3H)-yl)piperidine-2,6-dione (157 2(3H)-yl)piperidine-2,6-dione (157 mg, mg,0.4 0.4mmol) mmol)in in CHC13 CHCl (5 mL) (5 mL) was added was added TMSI TMSI (143 mg,(143 mg, 0.7 0.7 mmol), andthe mmol), and the mixture mixturewas wasstirred stirred at at room temperatureovernight. room temperature overnight.Then Then themixture the mixturewaswas washed bysat. washed by sat. NaHSO NaHSO3 (5 xmL (5 mL 2), washed 2),x washed by brine by brine (5 mL(5X mL 2), dried 2), xdried over over anhydrous anhydrous sodiumsodium

sulfate, sulfate,filtered and filtered concentrated. and The concentrated. Theresidue residuewas waspurified purifiedbybyPrep-TLC (DCM/MeOH Prep-TLC (DCM/MeOH = 15/1) = 15/1)

to to give give 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)piperidine-2,6-dione 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-alpyridin-2(3H)-yl)piperidine-2,6-dione

(130 mg, 79% (130 mg, 79% yield)asasa awhite yield) whitesolid. solid. LCMS

[0692] LCMS

[0692] LCMSLCMS (Agilent (Agilent 1200-6120, 1200-6120, Column: WatersWaters Column: X-Bridge X-Bridge C18 (50 4.6 xmm4.6 C18mm(50x mm mm x 3.5 m); Column 3.5 µm); ColumnTemperature: Temperature: 40 40 °C; °C; Flow Flow Rate: Rate: 2.0 2.0 mL/min; mL/min; Mobile Mobile Phase:Phase: from 95% from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 100%, Rt==1.754 100%, Rt 1.754min. min.MSMS Calcd.: Calcd.: 458.05; 458.05; MS MS Found: Found: 459.1459.1 [M+H]*.

[M+H].

[0693] Step Step

[0693] 6: Synthesis 6: Synthesis of N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yloxy)pentyl)piperazin-1-yl)nicotinamide

[1,2,4]triazolo[4,3-a]pyridin-7-yloxy)pentyl)piperazin-1-yl)nicotinamide

273

Oct 2023 IZ

NH NCNH H N H o ZI N N N N H N N N N N CI O O N N o DIEA, CHCN, 80 °C, o/n 2023248067 10

34% N N--N N-N o 34% IZ N NH N NN O N O - 0' O" CI CI

[0694] A solution

[0694] A solution of 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H) of 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-

yl)piperidine-2,6-dione (85 mg, yl)piperidine-2,6-dione (85 0.2 mmol), mg, 0.2 mmol),N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4- N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide (87 tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide (87mg, mg,0.20.2mmol), mmol), andand

ethyldiisopropylamine(72 ethyldiisopropylamine (72mg, mg,0.6 0.6mmol) mmol)in in acetonitrile(5(5 mL) acetonitrile mL)was was stirredatat 8080°C°Covernight. stirred overnight. When When itit was wascooled cooledtotoroom roomtemprature, temprature,water water (5 (5mL) mL) waswas added added and the and the mixture mixture was extracted was extracted

by ethyl by ethyl acetate acetate (5 (5mL mL X 3) and x 3) and the the combined organiclayers combined organic layerswere werewashed washedby by brine brine (5 (5 mL mL x 3), x 3),

dried dried over over anhydrous sodiumsulfate, anhydrous sodium sulfate, filtered filtered and and concentrated. concentrated. The residue was The residue was purified purified by by Prep-HPLC Prep-HPLC to to giveN-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4- give N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4 (5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7 (5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-

yloxy)pentyl)piperazin-1-yl)nicotinamide (50 yloxy)pentyl)piperazin-1-yl)nicotinamide mg,34%34% (50mg, yield) yield) as as a white a white solid. solid.

LC-MS

[0695] LC-MS

[0695] (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18mm(50x mm C18 (50 4.6 x 4.6 mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 3.0 in 3.0 min, min, then then under under thiscondition this conditionforfor1.0 1.0min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH0.1 3 CN] min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 100%, Rt==2.877 100%, Rt 2.877min; min;MSMS Calcd.: Calcd.: 797.34; 797.34; MS MS Found: Found: 798.3798.3 [M+H]*.

[M+H].

[0696] HPLCHPLC

[0696] (Agilent (Agilent HPLC 1200,1200, HPLC Column: Column: Waters Waters X-Bridge X-Bridge C18 (150 C18 (150 mm xmm 4.6xmm 4.6xmm 3.5x 3.5 pm); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 1.0 mL/min; 1.0 mL/min; MobileMobile Phase:Phase: from from 95% 95%+[water

[water 10 + 10 mMNH4HCO] mM NH 4HCOand 5%5% 3] and [CH 3 CN]

[CHCN] to 0% to 0% [water++10

[water 10 mM mMNH4HCO] NH 4HCOand 3] and 100% 100% [CH 3 CN]

[CHCN] in 10 in 10 min, then min, then under under this this condition condition for for 55 min, min, finally finallychanged changed to to95% [water ++ 10 95% [water 10 mM mM NH 4HCO 3] NH4HCO]

and 5%[CHCN] and 5% [CH 3 CN] in 0.1 in 0.1 min min and under and under this this condition condition for for 5 min.) 5 min.) Purity Purity is 93.85%, is 93.85%, Rt =Rt9.967 = 9.967

min. min.

(400 (400 'H NMR

[06971 ¹H NMR

[0697] MHz,MHz, DMSO-d) DMSO-d) 1.12 1.12 (6H, (6H, s), 1.21 1.21 (6H, s), (6H, s), 1.43-1.47 s), 1.43-1.47 (2H,m), (2H, 1.49 m),1.49- 1.53 (2H,m), 1.53 (2H, m),1.73-1.78 1.73-1.78 (2H,(2H, m), 2.13-2.17 m), 2.13-2.17 (1H, (1H, m), 2.32m), (2H,2.32 t, J(2H, = 7.2t,Hz), J= 2.43-2.47 7.2 Hz), (5H, 2.43-2.47 m), (5H, m),

274

2.61-2.62 (1H, (1H, m), m), 2.87-2.93 2.87-2.93 (1H, (1H, m), m), 3.59 3.59(4H, (4H,s), s), 4.01-4.07 4.01-4.07 (3H, (3H, m), m), 4.30 4.30 (1H, (1H, s), s), 5.28 5.28 (1H, (1H, dd, 2023248067 10 2023

2.61-2.62 dd, J= J 12.4, 5.2 = 12.4, 5.2Hz), Hz),6.35 6.35 (1H, (1H, dd, dd, J = J= 8.0, 8.0, 2.4 Hz), 2.4 Hz), 6.52 d,(1H, 6.52 (1H, d, Hz), J=1.6 J=1.66.86Hz), (1H, 6.86 (1H, d, J = 8.8 d, J= Hz), 8.8 Hz),

Oct 7.00 (1H,dd, 7.00 (1H, dd,J J= 8.8,2.42.4 = 8.8, Hz), Hz), 7.217.21 (1H,(1H, d, J d, J=Hz), = 2.4 2.4 7.63 Hz),(1H, 7.63d,(1H, d, J= J = 9.2 Hz),9.2 Hz), 7.80 (1H,7.80 d, J (1H, = d, J 8.0 Hz), 8.0 Hz), 7.90 7.90(1H, (1H,d, d, J =J= 8.88.8 Hz), Hz), 7.957.95 (1H, (1H, dd, J dd, J=2.4 = 9.2, 9.2,Hz), 2.48.62 Hz),(1H, 8.62 d, (1H, d, Hz), J = 2.4 J= 2.4 Hz), 11.09 11.09 (1H, s). (1H, s).

[0698] Chemical

[0698] Formula: C ChemicalFormula: CHCINO, 90, Molecular 4 1H 4 8 ClNMolecular Weight: Weight: 798.33. 798.33.

[0699]

[0699] Total H Total H count HNMR fromHNMR count from data: 48. 48. data:

[0700]

[0700] Exemplary Exemplary Synthesis of Exemplary Synthesis of Compound Exemplary Compound 49 49

O NH N N-ZO7 0 NNH N O N 0 NN O NNO N O N IZ H NC N N N CI CI 0 O Ó N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4-(5-((2-(2,6 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,24,4-tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-

dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-6 dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-I1,2,4]triazolo|4,3-a]pyridin-6-

yl)oxy)pentyl)piperazin-1-yl)nicotinamide yl)oxy)pentyl)piperazin-1-yl)nicotinamide

[0701] Synthetic

[0701] Scheme Synthetic Scheme

275

HN O Br Br H CI CI CI O-O CI N 2 'NH NH2 N N.~ N N. ~ -. N N 2H4 .H20 NH.HO . N N N HO HO 60% NaH, 60% NaHDMF DMF MW,170°C, MW, 170°C,1818 h h rt, 2Ihh rt, 2

0 0 o o NH NN NH NN N NH N NH NH Br Br =00 'NN N o CDI, CHCHCN CDI, 3 CN N.O NN N. O~-.- N N K2C0s CH 3CN 0 80°, KCO, CHCN 80 °C, 2 h o 80 °C, o/n o NH NH NN N 0H N HN o N NH N N IN N NC N CI O O o NNN TMSI, CHCI TMSI, CHCl o -l'

o rt, /n rt, o/n DIEA, CH CN,80 DIEA,CHCN, 80 °C,C, o/n 3o/n

o N NH N N o0 NNO N N 0 N ci~ N N N.NH 1N o N HN

N CI o Ó ~T Nt '-.

107021

[0702] Step 1: Step Synthesis of5-(5-(benzyloxy)pentyloxy)-2-chloropyridine 1: Synthesis of 5-(5-(benzyloxy)pentyloxy)-2-chloropyridine

SO Br Br O CI CI CI N HO N 60% NaH, NaH,DMF O O Ho 60% DMF rt,22 hh rt, 68% 68%

[0703]

[0703] Toa To solution of 6-chloropyridin-3-ol(1.0g,7.7mmol)inDMF(10mL)wasadded a solutionof 6-chloropyridin-3-ol (1.0 g, 7.7 mmol) in DMF (10 mL) was added

sodium hydride(60% sodium hydride (60% dispersed dispersed in in mineral mineral oil,371 oil, mg, 371mg, 9.3mmol) 9.3 at at0 mmol) 0 °C,°C, andthe and mixturewaswas themixture stirred stirredatat room roomtemperature temperature for for30mi. 30 min.Then Then ((5-bromopentyloxy)methyl)benzene ((5-bromopentyloxy)methyl)benzene (2.0 (2.0 g, 7.7 g, 7.7

mmol) was mmol)was added to to added thereaction the theresulted andthe reactionand wasstirred mixturewas resultedmixture at room stirredat room temperature for22h. temperaturefor h.

When thereaction When the wascompleted reactionwas completed (monitored (monitored by TLC), by TLC), water water (30 mL) mL)added. (30 was was added. The resultant The resultant

mixturewas mixture extractedbybyethyl wasextracted ethylacetate (10 mLx acetate (10 3) and mL x 3) the combined and the organiclayers combined organic layerswere were washed bybrine washed by brine(10 (10mL mLx x3), 3), dried over anhydrous dried over sodium anhydrous sodium sulfate, filtered and sulfate,filtered concentrated. and concentrated.

The residue (1.6 g,6800yield) was directly used to the next step without further purification as a The residue (1.6 g, 68% yield) was directly used to the next step without further purification as a

solid. brownsolid. brown

107041

[0704] Step Step 2: Synthesis 2: Synthesis of5-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine of 5-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine

276

2023248067 10 2023

H HN CI CI N'NH 2 NH O O NN N2H 4.H 20 , 0 N .O N NH.HO o O Oct MW, 170°C,18 MW,170°C, 18h h 82% 82%

[07051 To a To

[0705] microwave glass glass a microwave vial was was added vialadded 5-(5-(benzyloxy)pentyloxy)-2-chloropyridine 5-(5-(benzyloxy)pentyloxy)-2-chloropyridine

(1.6 (1.6 g, g, 5.2 5.2mmol) hydrazine monohydrate and hydrazine mmol) and monohydrate(20(20 mL), and and mL), the the mixture mixture was was stirred stirred under under

microwaveconditions microwave conditionsatat170 170°C°Cforfor1818h.h.When Whenit it waswas cooled cooled to to room room temperature, temperature, water water (20 (20

mL)was mL) wasadded added to to thereaction. the reaction. The Theresultant resultant mixture mixturewas wasextracted extractedbybyethyl ethylacetate acetate (10 (10 mL mLx x3)3) and the combined and the organiclayers combined organic layerswere werewashed washed by by brine brine (15(15 mL mL x 3), x 3), dried dried over over anhydrous anhydrous

sodium sulfate,filtered sodium sulfate, filteredandand concentrated concentrated in vacuo. in vacuo. The residue The residue (1.3yield) (1.3 g, 82% g, 82% was yield) was directly directly

usedtotothe used thenext nextstep stepwithout without further further purification purification as brown as brown oil. oil.

[0706] Step Step

[0706] 3: Synthesis 3: Synthesis of 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H) of 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-

one one H ZI N, N, N NH2 NH N NH NH CDI, CH CHCN CN ONO N N O O N CDI, 3 801& 80 °C, 22 h h O 19% 19%

[0707] To a To

[0707] a solution solution of 5-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine of 5-(5-(benzyloxy)pentyloxy)-2-hydrazinylpyridine (1.3 g,(1.3 4.4g,mmol) 4.4 mmol) in in acetonitrile acetonitrile(30 (30mL) mL) was was added CDI(1.1 added CDI (1.1g,g, 6.7 6.7 mmol), mmol),and andthe themixture mixturewas wasstirred stirredatat 80 80 °C °C for for 22 h. h.When it was When it was cooled to room cooled to temperature,water room temperature, water(20 (20mL) mL)waswas added added to the to the reaction.The reaction. The resultant mixture resultant mixture was extracted by was extracted ethyl acetate by ethyl acetate (10 (10 mL 3) and mL xx 3) and the the combined organiclayers combined organic layers were washed were washedbybybrine brine(15 (15mLmL x 3),dried x 3), driedover overanhydrous anhydrous sodium sodium sulfate, sulfate, filteredand filtered and concentrated in vacuo. concentrated in vacuo. The Theresidue residue was waspurified purified by bycolumn columnchromatography chromatography on silica on silica

(DCM/MeOH (DCM/MeOH = 20/1) = 20/1) to give to give 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H) 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-

one (280 mg, one (280 mg,19% 19% yield)asasa awhite yield) whitesolid. solid.

[0708] LC-MS

[0708] LC-MS (Agilent (Agilent LCMSLCMS 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18mm(50x mm C18 (50 4.6 x 4.6

mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO 3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed 95% changedtoto95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 98.98%, Rt==1.728 98.98%, Rt 1.728min. min.MSMS Calcd.: Calcd.: 327.16; 327.16; MS MS Found: Found: 328.1328.1 [M+H]*.

[M+H].

277

[0709] Step Step

[0709] 4: Synthesis 4: Synthesis of 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3 of 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-

a]pyridin-2(3H)-yl)piperidine-2,6-dione a]pyridin-2(3H)-yl)piperidine-2,6-dione

0 0 o N N O N NH N NH NH NH Br Br O'N o NH o O N N N o O K2 CO 3, CHCN CH3 CN O O KCO, O 0 o 80 °C, o/n 80 °C, o/n 41% 41%

2023248067

[0710] The solution

[0710] The solution of 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one of 6-(5-(benzyloxy)pentyloxy)-[1,2,4]triazolo[4,3-alpyridin-3(2H)-one

(280 mg, 0.9 (280 mg, 0.9 mmol), mmol),3-bromopiperidine-2,6-dione 3-bromopiperidine-2,6-dione (438 (438 mg, mg, 2.3 2.3 mmol) mmol) and(253 and KCO K2 C03 mg, (253 1.8 mg, 1.8

mmol)ininacetonitrile mmol) acetonitrile (10 (10 mL) wasstirred mL) was stirred at at 80 °C overnight. When °C overnight. When itit was wascooled cooledtotoroom room temperature, water (10 temperature, water (10 mL) mL)was wasadded. added.TheThe resultantmixture resultant mixture was was extracted extracted by by ethyl ethyl acetate(10 acetate (10 mLX x3)3) and mL andthe the combined combined organic organic layerswere layers were washed washed by brine by brine (10 (10 mL xmL 3), dried 3),x dried overover

anhydrous sodium anhydrous sodium sulfate,filtered sulfate, filtered and concentrated. The The residue residue was waspurified purified by by Prep-TLC Prep-TLC (DCM/MeOH = 20/1) (DCM/MeOH = 20/1) to give to give 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-

a]pyridin-2(3H)-yl)piperidine-2,6-dione (155 a]pyridin-2(3H)-yl)piperidine-2,6-dione (155 mg, mg,41% 41% yield) yield) as as a awhite whitesolid. solid.

[0711] LC-MS

[0711] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 4.6 x 4.6 C18mm(50x mm mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 85.76%,RtRt==1.675 85.76%, 1.675min. min.MSMS Calcd.: Calcd.: 438.19; 438.19; MS MS Found: Found: 439.2439.2 [M+H]*.

[M+H].

[0712] Step Step

[0712] 5: Synthesis 5: Synthesis of 3-(6-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin of 3-(6-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-

2(3H)-yl)piperidine-2,6-dione 2(3H)-yl)piperidine-2,6-dione

0 0 NNH o o N N NH TMSI, CHCs TMSI, CHCI N, NH 0'NNT0 NN HO N N O O N rt,o/n, rt, o/n,79% 79% |N N O 0 O 0

[0713] To a To

[0713] a solution solution of 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin of 3-(6-(5-(benzyloxy)pentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-

2(3H)-yl)piperidine-2,6-dione (155 2(3H)-yl)piperidine-2,6-dione (155 mg, mg,0.4 0.4mmol) mmol)in in CHC13 CHCl (5 mL) (5 mL) was added was added TMSI TMSI (143 mg,(143 mg, 0.7 0.7 mmol), andthe mmol), and the mixture mixturewas wasstirred stirred at at room temperatureovernight. room temperature overnight.Then Then themixture the mixturewaswas washedbybysat. washed sat. NaHSO NaHSO3 (5 xmL (5 mL 2), washed 2),x washed by brine by brine (5 mL(5X mL 2), dried 2), xdried over over anhydrous anhydrous sodiumsodium

sulfate, sulfate,filtered filteredand concentrated. and The concentrated. Theresidue residuewas waspurified purifiedbybyPrep-TLC (DCM/MeOH Prep-TLC (DCM/MeOH = 15/1) = 15/1)

to give to give 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)piperidine-2,6-dione 3-(7-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-alpyridin-2(3H)-yl)piperidine-2,6-dione

(130 mg, mg, 79% 79% yield)asasa awhite yield) whitesolid. solid.

278

1200-6120, Column: Waters X-Bridge 4.6 x 4.6 C18mm(50x mm 10 Oct 2023

LC-MS

[0714] LC-MS

[0714] (Agilent LCMSLCMS (Agilent 1200-6120, Column: Waters X-Bridge C18 (50

mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3 CN]to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3] and NH4HCO] and 100% 100%

[CH 3CN]

[CHCN] in 1.6 in 1.6 min, min, then then under under thiscondition this condition for1.4 for 1.4min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH4 HCO ] and and 35% 5% in

[CHCN] [CH 3CN] 0.1 min in 0.1under and and under min this this condition condition for 0.7 for 0.7 min.) min.) Purity isPurity is 95.44%,RtRt==1.706 95.44%, min.MSMS 1.706min. Caled.: Calcd.: 458.05; 458.05; MS MS 459.1459.1 Found: Found: [M+H]*.

[M+H]. 2023248067

[0715] Step Step

[0715] 6: Synthesis 6: Synthesis of N-((r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro tetramethylcyclobutyl)-6-(4-(5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-6-yloxy)pentyl)piperazin-1-yl)nicotinamide

[1,2,4]triazolo[4,3-a]pyridin-6-yloxy)pentyl)piperazin-1-yl)nicotinamide 0 $NHNH N NN NH NH N N 0 NN H IZ |NNO N N N NH NH N N Nf -NN N N N N CI N0 N H~o IZ

I -~O N N~, O C ON -NN 0 o DIEA, CHCN, CH 3CN, 80 o/n C, on CI DIEA, 80 °C, CI " .. 0 39% 39%

[0716] A solution

[0716] A solution of 3-(6-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H) of 3-(6-(5-iodopentyloxy)-3-oxo-[1,2,4]triazolo[4,3-alpyridin-2(3H)

yl)piperidine-2,6-dione (85 yl)piperidine-2,6-dione (85 mg, 0.2 mmol), mg, 0.2 mmol),N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4- N-((r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 tetramethylcyclobutyl)-6-(piperazin-1-yl)nicotinamide (87 (87mg, mg,0.2 0.2mmol), mmol),andand

ethyldiisopropylamine(72 ethyldiisopropylamine (72mg, mg,0.6 0.6mmol) mmol)in in acetonitrile(5(5 mL) acetonitrile mL)was was stirredatat80 stirred 80°C°Covernight. overnight. When When itit was wascooled cooledtotoroom roomtemprature, temprature,water water (5 (5 mL) mL) waswas added added and the and the mixture mixture was extracted was extracted

by ethyl by ethyl acetate acetate (5 (5mL mL x 3) and x 3) and the combined organiclayers combined organic layerswere werewashed washedby by brine brine (5 (5 mL mL x 3), x 3),

dried over anhydrous sodiumsulfate, anhydrous sodium sulfate, filtered, filtered, and and concentrated. concentrated. The The residue was purified by was purified by

Prep-HPLC Prep-HPLC to to giveN-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4- give N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-6-(4 (5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-6 (5-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo|4,3-a]pyridin-6-

yloxy)pentyl)piperazin-1-yl)nicotinamide (58 yloxy)pentyl)piperazin-1-yl)nicotinamide (58mg, mg,39%39% yield) yield) as as a white a white solid. solid.

[0717] LC-MS

[0717] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50x mm C18mm 4.6 x 4.6

mmx x3.5 mm 3.5µm); pm);Column Column Temperature: Temperature: 40 Flow 40 °C; °C; Flow Rate: Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010

[water mMmMNH 4HCO3] and NH4HCO] and 5% 5% [CH 3CN] to

[CHCN] to 0% [water + +1010 0% [water mMmMNH 4HCO 3] and NH4HCO] and 100% 100%

[CH 3CN]

[CHCN] in 3.0 in 3.0 min, min, then then under under thiscondition this condition for1.0 for 1.0min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH4 HCO ] and and 35% 5% in

[CHCN] [CH 3CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min.) min.) Purity is Purity is 100%, Rt==2.890 100%, Rt 2.890min; min;MSMS Caled.: Calcd.: 797.34; 797.34; MS MS Found: Found: 798.3798.3 [M+H]*.

[M+H].

HPLC

[0718] HPLC

[0718] (Agilent (Agilent HPLCHPLC 1200,1200, Column: Column: Waters Waters X-Bridge X-Bridge C18 (150 C18 (150 mm xmm 4.6xmm 4.6x mm 3.5x 3.5 pim); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 1.0 mL/min; 1.0 mL/min; MobileMobile Phase:Phase: from from 95% 95%+[water

[water 10 + 10 mMNH4HCO] mM NH4 HCO 3] and and 5%5% [CH 3CN]

[CHCN] to 0% to 0% [water++10

[water 10 mM mMNH4HCO] NH 4HCOand 3] and 100% 100% [CH 3CN]

[CHCN] in 10 in 10

279

Oct 2023 min, then min, then under under this this condition condition for for 55 min, min, finally finallychanged to95% changed to [water ++ 10 95% [water 10 mM mM NH 4HCO 3] NH4HCO]

and 5%[CHCN] and 5% [CH 3 CN] in 0.1 in 0.1 min min and under and under this this condition condition for for 5 min.) 5 min.) Purity Purity is 93.46%, is 93.46%, Rt =Rt10.027 = 10.027

min. min.

[0719]

[07191 ¹H NMR (400 (400 'H NMR MHz,MHz, DMSO-d) DMSO-d) 1.12 1.12 (6H, (6H, s), 1.21 1.21 (6H, s), (6H, s), 1.44-1.48 s), 1.44-1.48 (2H,m), (2H, 1.52 m),1.52- 2023248067 10

1.58 (2H,m), 1.58 (2H, m),1.74-1.79 1.74-1.79 (2H,(2H, m), 2.15-2.19 m), 2.15-2.19 (1H, (1H, m), 2.30m), (2H,2.30 t, J(2H, = 7.2t,Hz), J= 2.43-2.50 7.2 Hz), (4H, 2.43-2.50 m), (4H, m), 2.51-2.67(2H, 2.51-2.67 (2H, m),m), 2.86-2.95 2.86-2.95 (1H, (1H, m),(4H, m), 3.60 3.60s),(4H, 3.97s), 3.97 (2H, t, J(2H, t, Hz), = 6.4 J= 6.4 4.05Hz), (1H, 4.05 d, J =(1H, 9.2 d, J= 9.2 Hz), 4.30 Hz), 4.30(1H, (1H,s),s),5.38 5.38 (1H, (1H, dd, dd, J = J= 5.2,5.2, 12.8 12.8 Hz), Hz), 6.86d,(1H, 6.86 (1H, d, J= J = 9.2 Hz),9.2 Hz), 7.00 (1H,7.00 dd, (1H, dd, J = 8.4, J= 8.4, 2.4 Hz), 2.4 Hz), 7.10 7.10(1H, (1H,dd,dd, J =J= 10.0, 10.0, 2.0 2.0 Hz),Hz), 7.21 7.21 (1H, (1H, d, J = d, J= 2.4 2.47.25 Hz), Hz), 7.25 (1H, d, J(1H, d, J= = 10.0 Hz), 10.0 7.36 Hz), 7.36 (1H, s), 7.62 (1H, s), 7.62(1H, (1H,d,d,J J= = 9.29.2 Hz), Hz), 7.90 7.90 (1H,(1H, d, J d, J=Hz), = 8.8 8.8 7.95 Hz), (1H, 7.95dd, (1H, J = dd, 9.2,J= 2.4 9.2, Hz), 2.4 8.62Hz), 8.62

(1H, d, JJ= (1H, d, 2.4Hz), = 2.4 Hz),11.10 11.10 (1H,(1H, s). s).

[0720] Chemical

[0720] Formula: C ChemicalFormula: CHCINO, 90, Molecular 4 1H 4 8 ClNMolecular Weight: Weight: 798.33. 798.33.

[0721] Total

[0721] Total H count HNMR fromHNMR H countfrom data: 48.48. data:

[0722] Exemplary

[0722] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 50 Compound 50

N 0O 0 0 o O "N IZ N N O H N N 0 O N CI CI N N N NH N N N NH N 0 O N N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-((4-(2-(2,6 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(4-(2-(2,6-

dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1 dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-|1,2,4|triazolo[4,3-a|pyridin-7-yl)piperazin-1-

yl)methyl)piperidin-1-yl)benzamide yl)methyl)piperidin-1-yl)benzamide

[0723] Synthetic

[0723] Scheme Synthetic Scheme

280

Oct 2023 0 Br CI Ba-N Boc-N NH NH / CI CI NHNH 2 NHNH N N Br NH NH Br CI NH 2NH 2.H 20 CDI, CHCN HC NH Boc-N Boc N N N N NHNH+HO BoC N N /N N CDI, CH 3CN [N N NNH N Br N N NaO-tBu,Pd(dba) NaO-tBu, Pd (dba) N 120°C, 48 h Boc- reflux,1616h h Boc'NN Na-OtBu,CH3CN CH3 CN 2 120 °C, 48 h reflux, Na-OtBu, Xanthos,toluene Xantphos,toluene 22%yield 22% yield for for two two steps steps Boc reflux, 48 h reflux, 48 h

100 °C, 100 3h OC, 3h 23% 23% 46% 46%

2023248067 10 0

N N NH IN0 NJ 0 NNNH N N N TFA/DCM, rt, 2h H NN NNN =0 aB3C0 N, N0 IN N.O TFAIDCM, rt, 2 h N N') N NH=0 NH HN,_) HN o NH NeBHCN/ NaBHCN, N N N N Boc'0 Boc 98% MeOH/DCEIHOAc MeOH/DCE/HOAc O 98% rt, o/n

36% 36%

"NH2 0 0 NNH 0 O O CI o0 TFA, DCE HO IZ N N N EDCI, HOBt, DIPEA N H TFA3N rt., h N N N NN H NH EDC,HOBDPEA DMF, rt, o/n NI H CIN N N N NH 83% 83% N,) N 0 IDIMF,rt, o/n 52% NIN 1 52%

[0724]

[0724] Step 1:1: Synthesis Step Synthesisof of tert-butyl tert-butyl 4-(2-chloropyridin-4-yl)piperazine-1-carboxylate 4-(2-chloropyridin-4-yl)piperazine-1-carboxylate CI Br Br CI CI Boc-N Boc N NH NH CI

N ,z.. N NaO-tBu, Pd NaO-tBu, 2(dba) 3 Pd(dba) Boc-N Boc-N N N NN Xantphos,toluene Xantphos, toluene 100 OC, 33 hh 100 °C, 46% 46%

[0725] To a To

[0725] a solution solution of 4-bromo-2-chloropyridine of 4-bromo-2-chloropyridine (5.830.2 (5.8 g, g, 30.2 mmol)mmol) in dryin dry toluene toluene (150 (150 mL)was mL) wasadded added sodium sodium tert-butoxide tert-butoxide (4.3 (4.3 g, g,45.0 45.0mmol), mmol), Pd 2 (dba) Pd(dba) 3 (0.55 (0.55 g, 0.60 g, 0.60 mmol), mmol),

Xantphos(1.0 Xantphos (1.0 g, g, 1.80 1.80 mmol) mmol)and andtert-butyl tert-butyl piperazine-1-carboxylate piperazine-1-carboxylate(5.6 (5.6 g,g, 30.2 30.2 mmol). mmol).The The reaction mixture reaction mixturewaswas stirred stirred at 100 at 100 °C3for °C for 3 h under h under nitrogen nitrogen and thenand then cooled to cooled rt. The to rt. The organic organic layer layer was washedwith was washed withwater waterand andbrine brineand andthen thendried driedover overanhydrous anhydrous sodium sodium sulfate, sulfate, filtered, filtered,

and concentrated. The and concentrated. Theresidue residue was waspurified purified by by silica silica gel gel chromatography column chromatography column (PE/EA (PE/EA = 8:1) = 8:1)

to give to give tert-butyl tert-butyl 4-(2-chloropyridin-4-yl)piperazine-1-carboxylate 4-(2-chloropyridin-4-yl)piperazine-1-carboxylate (3.6asg,a46%) (3.6 g, 46%) yellow as a yellow solid. solid.

[0726]

[0726] ¹H 'H NMR NMR (400 MHz, (400 DMSO-d) MHz, 1.42 (9H, DMSO-d) s), (9H, 6 1.42 3.38-3.41 (8H, m),(8H, s), 3.38-3.41 6.83-6.86 (2H, m), (2H, m), 6.83-6.86 m), 7.96 (1H,d,d,J J= 7.96 (1H, = 6.0 Hz). 6.0Hz). Formula: C ChemicalFormula:

[0727] Chemical

[0727] CHCINO, 3 0 2 , Molecular 14 H 2 0 ClNMolecular Weight: Weight: 297.78. 297.78.

[0728]

[0728] Total H Total H count HNMR fromHNMR count from data: 20. 20. data:

[0729]

[0729] Step 2: Step 2: Synthesis Synthesis of of tert-butyl tert-butyl4-(2-hydrazinylpyridin-4-yl)piperazine-1-carboxylate 4-(2-hydrazinylpyridin-4-yl)piperazine-1-carboxylate

281

CI CI NHNH 2 NHNH -- NH 2 NH 2•H 2 0 NHNH·HO Boc-N - /C N Boc-N Boc-N N N / NN ___Boc-N__NN Boc-N N N 120 °C, 120 48hh °C, 48

[0730]

[0730] To To aasolution solutionofof tert-butyl4-(2-chloropyridin-4-yl)piperazine-1-carboxylate tert-butyl 4-(2-chloropyridin-4-yl)piperazine-1-carboxylate (5.0 g, (5.0 g, 16.8 16.8 mmol) mmol) ininhydrazine hydrazinemonohydrate monohydrate (98%, (98%, 40 mL), 40 mL), was stirred was stirred at 120 at 120 °C for °C for 48 h48under h under nitrogen. Water nitrogen. (100 mL) Water (100 mL)was wasadded added to to thethe mixture.TheThe mixture. resultantmixture resultant mixture was was extracted extracted by by ethyl ethyl

2023248067

acetate acetate (50 (50 mL 3), washed mL xx 3), bybrine washed by brine(100 (100 mL), mL),dried driedover overanhydrous anhydrous sodium sodium sulfate, sulfate, filtered, filtered,

and concentrated and concentrated in vacuo. in vacuo. The residue The residue (4.8 g,(4.8 30% g, 30% was purity) purity) wasused directly directly to theused next to the next step step

withoutfurther without furtherpurification purification as aasbrown a brown solid.solid.

[0731] Step Step

[0731] 3: Synthesis 3: Synthesis of tert-butyl of tert-butyl 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-

yl)piperazine-1-carboxylate yl)piperazine-1-carboxylate

0 o NHNH 2 NN NH NHNH NH CDI, CHCN N N N Boc-N N-/"N CDI, CH 3CN N N Boc-N N N reflux, 16 h reflux, 16 h Boc' Boc N 22%yield 22% yieldfor fortwo twosteps steps

[0732] To a To

[0732] a solution solution of tert-butyl of tert-butyl 4-(2-hydrazinylpyridin-4-yl)piperazine-1-carboxylate(4.8(4.8 14-(2-hydrazinylpyridin-4-yl)piperazine-1-carboxylate

g, g, 30% purity, 4.9 mmol) 30% purity, in acetonitrile mmol) in acetonitrile (100 (100 mL) wasadded mL) was addedCDI CDI (1.6g, g,9.8 (1.6 9.8mmol), mmol),andand thethe

mixture stirred mixture stirred atat 100 100 °C °C for for 16 16 h.h.When it was When it was cooled to room cooled to temperature, water room temperature, water(100 (100mL) mL) was was

added to the added to the reaction. reaction. The The resultant resultantmixture mixture was was extracted extracted by by ethyl ethyl acetate acetate(100 (100 mL mL X 3), washed x 3), washed

by brine by brine (150 mL), dried (150 mL), dried over over anhydrous anhydroussodium sodium sulfate,filtered, sulfate, filtered, and and concentrated concentrated in in vacuo. vacuo. The The residue was residue purified by coloumn was purified coloumnchromatography chromatography on silica on silica (DCM/MeOH (DCM/MeOH = 20/1) =to20/1) give to give tert- tert butyl 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate butyl 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate(1.2 g, (1.2 g, 22%yield 22% yield forfor twotwo steps) steps) as aas a yellow yellow solid.solid.

[0733] LC-MS

[0733] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[(total 10mM

[(total 10mM AcONH AcONH4)4)water/CHCN=900/100 water/CH3CN=900/100 (v/v)] (v/v)] andand10%10% [(total 10mM

[(total 10mM AcONH 4 AcONH4) )

water/CH3CN=100/900 (v/v)]toto 10% water/CHCN=100/900 (v/v)] 10%[(total

[(total 10mM AcONH water 10mM AcONH4) 4) water /CH 3 CN=900/100 /CHCN=900/100 (v/v)] (v/v)]

and 90%[(total and 90% [(total 10mM 10mM AcONH AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] (v/v)] in 1.6 in then min, 1.6 min, underthen under this this

condition for for 2.4 2.4 min, min,finally finallychanged changed to to 90% [(total10mM 90% [(total AcONHwater/CH3CN=900/100 10mM AcONH4) 4) water/CH3CN=900/100

(v/v)] (v/v)] and and 10% [(total10mM 10% [(total AcONHwater/CHCN=100/900 10mM AcONH4) 4) water/CH3CN=100/900 (v/v)] in (v/v)] 0.1 minin and 0.1 under min and under this this

condition for for 0.7 0.7 min). min). Purity Purity isis99.11%, 99.11%, Rt Rt == 1.418 1.418 min; min; MS Calcd.: 319.7; MS Calcd.: 319.7;MSMSFound: Found: 320.2 320.2

282

[M+H]*. 2023248067 10 2023

[M+H].

[07341 ¹H 'H NMR NMR (400 MHz, (400 DMSO-d) MHz, 1.42 6(9H, DMSO-d s), (9H, ) 6 1.42 3.21-3.23 (4H, m),(4H, s), 3.21-3.23 3.42-3.43 (4H, m), m), 3.42-3.43

[0734] (4H, m),

Oct 6.13 (1H,d,d,J J= 6.13 (1H, 1.6Hz), = 1.6 Hz), 6.60 6.60 (1H,(1H, dd, Jdd, J= 2.0 = 8.0, 8.0,2.0 Hz),(1H, Hz), 7.65 7.65d,(1H, d, J= J = 8.0 Hz),8.0 Hz), 11.90 11.90 (1H, s). (1H, s).

[0735] Chemical

[0735] Formula: C ChemicalFormula: 1 5 H 2 1N CHNO, 5 03 , Molecular Molecular Weight: Weight: 319.36 319.36

[0736]

[0736] Total H count Total H HNMR fromHNMR count from data: 21. 21. data:

[0737]

[0737] Step 4: Step 4: Synthesis Synthesisof of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro- tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate

0 o o0 0 o -- NH NH N NH Br Br 0 N N NH N N N N IN N N,N O Na-OtBu, CH N NH N N Na-OtBu, 3 CN CHCN BocNN NH BocN Boc reflux, 48 h reflux, 48 h Boc 0 o 23% 23%

The solution

[0738] The solution

[0738] of tert-butyl of tert-butyl 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7 4-(3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-

yl)piperazine-1-carboxylate (320 yl)piperazine-1-carboxylate (320 mg, mg,1.0 1.0mmol), mmol),3-bromopiperidine-2,6-dione 3-bromopiperidine-2,6-dione (390(390 mg, mg, 2.0 2.0 mmol)and mmol) andsodium sodium tert-butoxide tert-butoxide (120 (120 mg,mg, 1.21.2 mmol) mmol) in acetonitrale in acetonitrale (20(20 mL)mL) was was stirred stirred at at 100 °C overnight. 100 °C overnight. When Whenititwas wascooled cooledtotoroom room temperature, temperature, water water (20(20 mL)mL) was was added. added. The The

resultant mixture resultant mixture was extracted by was extracted ethyl acetate by ethyl acetate (20 (20 mL 3), washed mL xx 3), bybrine washed by brine (30 (30 mL), mL),dried dried over anhydrous anhydroussodium sodium sulfate,filtered sulfate, filtered and concentrated. The and concentrated. The residue residue was waspurified purified by by Prep-TLC Prep-TLC (DCM/MeOH = 20/1) (DCM/MeOH = 20/1) to give to give tert-butyl tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate (100

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate (100 mg, 23%yield) mg, 23% yield) asas aa yellow yellow solid. solid.

[0739]

[0739] ¹H 'H NMR NMR (400 MHz, (400 DMSO-d) MHz, 1.47 (9H, DMSO-d) s), (9H, 6 1.47 2.16-2.20 (1H, m),(1H, s), 2.16-2.20 2.47-2.50 (1H, m), (1H, m), 2.47-2.50 m), 2.66-2.70 (1H, m), 2.66-2.70 (1H, m), 2.90-2.99 (1H, m), 2.90-2.99 (1H, m), 3.31-3.33 3.31-3.33 (4H, (4H,m), m),3.47-3.49 3.47-3.49(4H, (4H,m), m),5.27-5.31 (1H, 5.27-5.31(1H, m), m),

6.20 (1H,d,d,J J= 6.20 (1H, 1.2Hz), = 1.2 Hz), 6.73 6.73 (1H,(1H, dd, Jdd, J= 2.0 = 7.6, 7.6, Hz), 2.0 7.80 Hz),(1H, 7.80d,(1H, d, J= J = 8.0 Hz),8.0 Hz), 11.11 11.11 (1H, s). (1H, s).

[0740] Chemical

[0740] Formula: C ChemicalFormula: 20 H 2 6N CHNO, 60, Molecular Molecular Weight: Weight: 430.46. 430.46.

[0741]

[0741] Total H count Total H count from HNMR fromHNMR data: 26. 26. data:

[0742] Step Step

[0742] 5: Synthesis 5: Synthesis of 3-(3-oxo-7-(piperazin-1-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H) of 3-(3-oxo-7-(piperazin-1-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-

yl)piperidine-2,6-dione yl)piperidine-2,6-dione

0 O O N NN N N N TFA/DCM, rt, 2 h N Bc N' N O TFA/DCM, rt,2h N HN N ) BoN N N' NH HN ,) HN NH NH O Boc NBo-N0O 98% 98%0

[0743]

[0743] To To aasolution solutionofof tert-butyl4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro- tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro

283

2023 [1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazine-1-carboxylate (0.40 g,(0.40 g, 0.93 0.93 mmol) in mmol) in dichloromethane (20mL) dichloromethane (20 mL)waswas added added TFA TFA (8 mL), (8 mL), then then stirred stirred at room at room temperature temperature for 2for 2 h and h and

2023248067 10 Oct

concentrated in vacuo concentrated in vacuoto to give give 3-(3-oxo-7-(piperazin-l-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H)- 3-(3-oxo-7-(piperazin-1-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H) (0.30 g, yl)piperidine-2,6-dione (0.30 yl)piperidine-2,6-dione g, 98%) as aa yellow 98%) as solid, which yellow solid, which was to the usedto was used the next next step step without without

further purification. further purification.

[0744] Step Step

[0744] 6: Synthesis 6: Synthesis of tert-butyl of tert-butyl 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3 4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-

dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate

o O o 0 N /N N N-'NH N NH N 0 N 0 N o N N N' N N 0 (N N HN N N' NH O NaBH 3CN, / O \/N N HN NaBHCN, O N O MeOH/DCE/HOAc N MeOH/DCE/HOAc rt, o/n rt, o/n

36% 36%

[0745] To a To

[0745] a solution solution of 3-(3-oxo-7-(piperazin-1-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H) of 3-(3-oxo-7-(piperazin-1-yl)-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-

yl)piperidine-2,6-dione (0.30 yl)piperidine-2,6-dione (0.30 g, g, 0.91 0.91 mmol) in dry mmol) in dry methanol/1,2-dichloroethane/HOAc methanol/1,2-dichloroethane/HOAc (20 (20 mL/4mL/4

mL/0.1mL) mL/0.1 mL)waswas added added tert-butyl4-(4-formylpiperidin-1-yl)benzoate tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (0.26 (0.26 g, g, 0.91 0.91 mmol). mmol). The The

mixture was mixture wasleft left to to stir stirfor 3030min for underNN2 gas. minunder gas. Then sodiumcyanoborohydride Then sodium cyanoborohydride (0.11 (0.11 g, 1.82 g, 1.82

mmol)was mmol) wasadded added andand thethe reactionmixture reaction mixture waswas lefttotostir left stir for for 16 16 hh at at room temperature. The room temperature. The solvent solvent was removedand was removed and theresidue the residuepartitioned partitionedbetween betweendichloromethane dichloromethane and and water, water, washed washed

withbrine, with brine,dried driedover over anhydrous anhydrous sodium sodium sulfate,sulfate, filtered, filtered, and concentrated and concentrated in vacuo toingive vacuo to give crude product. The crude product. residue was The residue waspurified purified by by prep-TLC prep-TLCto to givecompound give compound tert-butyl tert-butyl 4-(4-((4-(2 4-(4-((4-(2-

(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1 (2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-|1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-

yl)methyl)piperidin-1-yl)benzoate (0.20 yl)methyl)piperidin-1-yl)benzoate (0.20 g, g, 36%) 36%)asasaayellow yellowsolid. solid.

[0746] LC-MS

[0746] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5 pm); mm*3.5 µm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow Rate: 2.0 mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 90%from 90%

[(total 10mM

[(total 10mM AcONH AcONH4)4)water/CHCN=900/100 water/CH3CN=900/100 (v/v)] (v/v)] andand10%10% [(total 10mM

[(total AcONH 4 10mM AcONH4) )

water/CH3CN=100/900 (v/v)] toto 10% water/CHCN=100/900 (v/v)] 10%[(total

[(total 10mM AcONH water 10mM AcONH4) 4) water /CH 3 CN=900/100 r/CHCN=900/100 (v/v)] (v/v)]

and 90%[(total and 90% [(total 10mM 10mM AcONH AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] (v/v)] in 1.6 in then min, 1.6 min, underthen under this this

condition for 2.4 condition for 2.4 min, min, finally finallychanged changed to to 90% [(total10mM 90% [(total AcONH 10mM AcONH4) 4) water/CH3CN=900/100 water/CHCN=900/100

(v/v)] (v/v)] and and 10% [(total10mM 10% [(total AcONH 10mM AcONH4) 4) water/CH3CN=100/900 water/CHCN=100/900 (v/v)] in (v/v)] 0.1 minin and 0.1 under min and under this this

condition for for 0.7 0.7 min). min). Purity Purity isis87.07%, 87.07%, Rt Rt == 2.195 2.195 min.; min.; MS Calcd.: 603.3; MS Calcd.: 603.3; MS MSFound: Found: 604.4 604.4

[M+H]*.

[M+H].

[0747] Step Step

[0747] 7: Synthesis 7: Synthesis of 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro of 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

284

Oct 2023

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acidl acid 0 O N N o NH TFA, DCE N NN TFA, DCE Ho HO N NN O o o o N rt., 2 h NN N N N NH N rt 2 h O 83% o 2023248067 10 N N N

[0748] To a To

[0748] a solution solution of tert-butyl of tert-butyl 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro 4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (0.10 g, (0.10 0.16 g, 0.16 mmol)inindichloromethane mmol) dichloromethane(10(10 mL)mL) was was added added TFA TFA (5 (5 then mL), mL),stirred then stirred at room at room temperature temperature for for 22 h, h, then then concentrated concentrated in in vacuo vacuo to to give give 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro 4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-l-yl)methyl)piperidin-1-yl)benzoic acid acid. (0.075 g, (0.075 g, 83%)asasa ayellow 83%) yellow solid, solid, which which wastoused was used to the the next next step step further without withoutpurification. further purification.

[0749] Step Step

[0749] 8: Synthesis 8: Synthesis of N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 of N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro tetramethylcyclobutyl)-4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide

o 0q0O o NH 'NH N/ NH2 N N o OO o N CI N HO N o Ho CNN N 0 O N DIPEA HOBt, DIPEA EDCI, HOBt, N N N NH EDCI, N N N NH DMF, rt, o/n N N NN DMF, rto/n N N o ~ 52%0 52% N N O N

O -NH "NH o

CI CI / N N

[0750] A solution

[0750] A solution of 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro of 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid. (75 mg, (75 mg, 0.14 0.14

mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiinide mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride hydrochloride (EDCI) (EDCI) (390.21 (39 mg, mg, 0.21 mmol),1-hydroxybenzotriazole mmol), 1-hydroxybenzotriazolehydrate hydrate (HOBt) (HOBt) (28 (28 mg, mg, 0.21 0.21 mmol)mmol) and ethyldiisopropylamine and ethyldiisopropylamine

(88 mg, 0.69 mmol) mg, 0.69 mmol)ininDMF DMF (5 mL) (5 mL) was was stirred stirred for for 30 min, 30 min, and and thenthen 4-((r,3r)-3-amino-2,2,4,4 4-(1r,3r)-3-amino-2,2,4,4-

tetramethylcyclobutoxy)-2-chlorobenzonitrile (38 tetramethylcyclobutoxy)-2-chlorobenzonitrile (38mg, mg,0.14 0.14mmol) mmol) was was added. added. The The mixture mixture was was stirred stirredatatroom room temperature temperature overnight and water overnight and water (10 (10 mL) mL)was was added.TheThe added. aqueous aqueous layer layer was was

extracted extracted by by dichloromethane (20mLmL dichloromethane (20 x 2).The x 2). The combined combined organic organic layer layer was was washed washed by brine by brine (10 (10

mLx x2), mL 2), dried dried over over anhydrous anhydroussodium sodium sulfate,filtered, sulfate, filtered, and and concentrated in vacuo. concentrated in vacuo. The residue The residue

285

was purified was purified by prep-HPLC by prep-HPLC to to give give N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4

tetramethylcyclobutyl)-4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro tetramethylcyclobutyl)-4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-3-oxo-2,3-dihydro-

[1,2,4]triazolo[4,3-a]pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (57 mg,

[1,2,4]triazolo[4,3-a|pyridin-7-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (57 mg,52%) 52%) as a white as a solid. white solid.

[0751] LC-MS

[0751] LC-MS (Agilent LCMSLCMS (Agilent 1200-6120, 1200-6120, Column: Column: Waters Waters X-Bridge X-Bridge C18 (50 (50 mm*4.6 C18mm*4.6 mm*3.5µm); mm*3.5 pm); Column Column Temperature: Temperature: 40Flow 40 °C; °C; Rate: Flow 2.0 Rate:mL/min; 2.0 mL/min; Mobile Mobile Phase: Phase: from 95%from 95%

[water + +1010 mMmMNH 4HCO3] and and 5% 5% [CH 3 CN]to to 0% [water + +1010 2023248067

[water NH4HCO] [CHCN] 0% [water mMmMNH 4HCO 3 ] and NH4HCO] and 100% 100%

[CH 3 CN]

[CHCN] in 3.0 in 3.0 min, min, then then under under thiscondition this condition for1.0 for 1.0min, min,finally finally changed changedtoto95% 95% [water

[water + + 10 10 mMNH4HCO] mM NH 4HCO ] and and 35% 5% in

[CHCN] [CH 3 CN] 0.1 min in 0.1under and min this and under this condition condition for 0.7 for 0.7 min). min). Purity is Purity is 98.22%,RtRt==3.022 98.22%, 3.022min; min;MSMS Calcd.: Calcd.: 807.4; 807.4; MS MS Found: Found: 808.3808.3 [M+H]*.

[M+H].

HPLC

[0752] HPLC

[0752] (Agilent (Agilent HPLC 1200,1200, HPLC Column: Column: Waters Waters X-Bridge X-Bridge C18 (150 C18 (150 mm*4.6 mm*4.6 mm*3.5 mm*3.5

pm); Column µm); Column Temperature: Temperature: 40 °C; 40 °C; FlowFlow Rate: Rate: 1.0 mL/min; 1.0 mL/min; MobileMobile Phase:Phase: from from 95% 95%+[water

[water 10 + 10 mMNH4HCO] mM NH 4HCOand 5%5% 3] and [CH 3 CN]

[CHCN] to 0% to 0% [water++10

[water 10 mM mMNH4HCO] NH 4HCOand 3] and 100% 100% [CH 3 CN]

[CHCN] in 10 in 10 min, then min, then under under this this condition condition for for 55 min, min, finally finallychanged changed to to95% [water ++ 10 95% [water 10 mM mM NH 4HCO 3] NH4HCO]

and 5%[CHCN] and 5% [CH 3 CN] in 0.1 in 0.1 min min and under and under this this condition condition for for 5 min). 5 min). Purity Purity is 99.00%, is 99.00%, Rt =Rt10.305 = 10.305

min. min.

[0753]

[0753] ¹H 'H NMR NMR (400 MHz, (400 DMSO-d) MHz, 1.13 (6H, DMSO-d) s), (6H, 6 1.13 1.22 s), (6H, s),(6H, 1.22 s), 1.79-1.81 1.79-1.81 (3H, (3H, m), m), 2.09- 2.09 2.15 (1H, m), 2.15 (1H, m), 2.19-2.21 2.19-2.21 (2H, (2H, m), m), 2.49-2.50 2.49-2.50 (7H, (7H, m), m),2.60-2.67 (1H,m), 2.60-2.67(1H, m),2.76-2.92 2.76-2.92(3H, (3H,m), m),3.22- 3.22 3.26 (4H,m), 3.26 (4H, m),3.86 3.86 (2H, (2H, d, Jd,= J= 12.8 12.8 Hz), Hz), 4.05d,(1H, 4.05 (1H, d, J= J = 9.2 Hz),9.2 Hz), 4.32 (1H,4.32 s), (1H, s), 5.23 5.23 (1H, dd, J(1H, = dd, J= 12.4, 5.2 Hz), 12.4, 5.2 Hz),6.12 6.12(1H, (1H,s),s), 6.70 6.70 (1H, (1H, dd, dd, J = J= 8.0, 8.0, 1.6 Hz), 1.6 Hz), 6.95d,(2H, 6.95 (2H, d, J= J = 9.2 Hz),9.2 Hz), 7.00 (1H,7.00 dd, (1H, dd,

JJ= 8.8, 2.4 = 8.8, 2.4 Hz), Hz),7.21 7.21(1H, (1H, d,=J= d, J 2.4 2.4 Hz),Hz), 7.48 7.48 (1H, (1H, d, J =d,8.8 J= 8.87.72 Hz), Hz),(3H, 7.72 t, (3H, t, J= J = 8.4 Hz), 8.4 7.91Hz), 7.91 (1H, d, JJ= (1H, d, 8.8Hz), = 8.8 Hz),11.04 11.04 (1H,(1H, s). s).

[0754] Chemical

[0754] Formula: C ChemicalFormula: 4 3 H5 0 ClN CHCINO, 90, Molecular Molecular Weight: Weight: 808.37. 808.37. Total

[0755] Total

[0755] H count H count from from HNMR HNMR data: 50.50. data: General

[0756] General

[0756] Synthetic Synthetic Scheme C-3 C-3 Scheme

[0757] Synthesis

[0757] Synthesis of building of building blockblock N-((r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-4-(4-formylpiperidin-1-yl)benzamide tetramethylcyclobutyl)-4-(4-formylpiperidin-1-yl)benzamide

CI 'N N H H -O N"N N C O

286

Oct 2023

[0758]

[0758] Synthetic Scheme: Synthetic Scheme:

OH F HN HN S\O N N OH OH 1.37%NaOH HO O 1. 37% NaOH F F O / K2 C03 , DMSO Noo 2. HCI 2. HCI /~N N O O 00 KCO, DMSO O H OH O 2023248067 10

CI CI

NH 2 'NH C 0O CI N NHCI HCI N O 0 O 'N

HATU,DIEA, HATU, DIEA,DMF, 30 °C, 70 min N H DMF, 30 °C, 70 min N No", OH OH

0 CI CI N o 0 o Dess-Martin Dess-Martin "NO N DCM,3030°C, DCM, °C,2h 2h N N H H N N '-0 O

[0759]

[0759] 1: Synthesis Step 1: Step of Ethyl Synthesis of 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate Ethyl4-(4-(hydroxymethyl)piperidin-1-yl)benzoate

OH 0-o HNC I" OH O HN N OH F F K 2CS O 3 , DMSO KCO, DMSO O O

[0760] To a To

[0760] a solution solution of ethyl of ethyl 4-fluorobenzoate 4-fluorobenzoate (270.16 (27 g, g, 0.16 mol)mol) in DMSO in DMSO (500 (500 mL) wasmL) was added K2 C03 added KCO (440.32 (44 g, g, 0.32 mol)mol) and piperidin-4-ylmethanol and piperidin-4-ylmethanol (320.19 (32 g, g, 0.19 mol) mol) at 25°C. at 25°C. The The

resulting solution resulting solution was was stirred stirredatat100 100°C°Cfor for1212h. h. The reaction The was reaction wasdiluted dilutedwith withH 2HO 0 (600 (600 mL). mL). The resulting mixture The resulting wasextracted mixture was extracted with withEtOAc EtOAc (200 (200 mL mL x 3). X 3). TheThe combined combined organic organic layers layers

were dried were dried over over anhydrous anhydroussodium sodium sulfateand sulfate and concentration.The concentration. The crude crude product product waswas slurry slurry in in PE/MTBE=1:1 PE/MTBE=1:1 to afford to afford ethyl ethyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (3071% (30 g, 71% yield) g, yield) as as aa white solid, which white solid, whichwaswas usedused into into next next step without step without further further purification. purification.

ChemicalFormula:

[0761] Chemical

[0761] Ci 5 H 2iNO Formula:CHNO; 3; Molecular Molecular Weight: Weight: 263.34. 263.34.

[0762] ¹H NMR

[0762] 'H NMR (400 (400 MHz, MHz, DMSO-d): DMSO-d): 6 7.91 7.91 (d, J = (d, 8.8J= 8.82H), Hz, Hz, 6.87 2H), 6.87 (d, J(d,=J= 8.88.8 Hz,2H), Hz, 2H), 4.30-4.35(m,(m,2H), 4.30-4.35 2H), 3.903.90 (d, (d, J = J= 12.8 12.8 Hz, 3.54 Hz, 2H), 2H),(d, 3.54 J = (d, 6.4J= Hz, 6.4 2H),Hz, 2H), 2.82-2.89 2.82-2.89 (m,(d, (m, 2H), 1.85 2H), 1.85 (d, J= J 12.8 Hz, = 12.8 2H), 1.71-1.77 Hz, 2H), 1.71-1.77 (m, (m, 1H), 1H), 1.35-1.54 1.35-1.54 (m, (m,6H). 6H).

[0763] Total

[0763] Total H count H count from NMRNMR ¹H IH from 21 21 data:data:

[0764] 2: 2:

[0764] StepStep Synthesis acid 4-(4-(Hydroxymethyl)piperidin-1-yl)benzoic acid Synthesisofof4-(4-(Hydroxymethyl)piperidin-1-yl)benzoic

287

Oct 2023 0 0 o o NaOH NaOH OH N N THF, MeOH, THF, H 2 0,30 MeOH, HO, 30 °C, 12h °C, 12h N N HO__HO Ho Ho

2023248067 10

[0765] To a To

[0765] a solution solution of ethyl of ethyl 4-[4-(hydroxymethyl)-1-piperidyl]benzoate 4-[4-(hydroxymethyl)-1-piperidyl]benzoate (52 g,(52 g, 197.47 197.47 mmol, mmol, 1 eq) eq) in in tetrahydrofuran tetrahydrofuran (250 (250 mL), mL), methanol (250mL) methanol (250 mL)andand water water (250 (250 mL)mL) was was addedadded sodium sodium

hydroxide (31.6 g, hydroxide (31.6 g, 0.79 0.79 mmol, mmol,4 4eq). eq). The Themixture mixturewas wasstirred stirredatat 30 30 °C °C for for 12 12 hours. hours. Thin Thinlayer layer chromatography (petroleum chromatography (petroleum ether:ethyl ether: ethylacetate=1:1) acetate=1:1)showed showed thethe reaction reaction was was completed. completed. The The

mixture was mixture wasadjusted adjustedtoto pH pH3~4 3-4with withhydrochloric hydrochloricacid acid (2(2M)M) andand filtered. The filtered. Thefilter filter cake cake was was dried in vacuum. dried in vacuum.TheThe residue residue was triturated was triturated withacetate with ethyl ethyl acetate (500 mL) (500 mL) to give to give 4-[4 4-[4-

(hydroxymethyl)-1-piperidyl]benzoicacid (hydroxymethyl)-1-piperidyl]benzoic acid (35g,g,148.76 (35 148.76mmol, mmol, 75% 75% yield) yield) as aaswhite a white solid. solid.

[0766] ¹H NMR:

[0766] 1H NMR: (400MHz, (400MHz, DMSO-d) DMSO-d) : 12.19 : 12.19 (s, 1H),(s,7.74 1H),(d, 7.74J=8.8 (d, J=8.8 Hz, Hz, 2H),2H), 6.936.93 (d,(d,

J=8.8Hz, J=8.8 Hz,2H), 2H), 4.48 4.48 (br (br t, J=5.2 t, J=5.2 Hz, 3.90 Hz, 1H), 1H), (d, 3.90 (d, J=12.8 J=12.8 Hz, 2H),Hz, 3.272H), 3.27 (br t, (brHz, J=5.2 t, J=5.2 Hz, 2H), 2.86 2H), 2.86 - 2.72 2.72 (m, (m, 2H), 2H), 1.72 1.72 (d, (d, J=12.8 Hz, 2H), J=12.8 Hz, 2H), 1.66- 1.66 -1.51 1.51(m, (m,1H), 1H),1.17 1.17(dq, (dq, J=4.0, J=4.0,12.0 12.0 Hz, Hz,2H) 2H) Formula: C ChemicalFormula:

[0767] Chemical

[0767] 13 H 17 NO CHNO, 3 , Molecular Molecular Weight: 235.28. Weight:235.28. Total

[0768] Total

[0768] H count H count from from HNMR HNMR data: 17.17. data:

[0769] Step Step

[0769] 3: Synthesis 3: Synthesis of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4- of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4- tetramethyl tetramethyl-

cyclobutyl]-4-[4-(hydroxymethyl)-1-piperidyl]benzamide cyclobutyl]|-4-|4-(hydroxymethyl)-1-piperidyl]benzamide

CI CI 0

o 'NH CI O N 'NH 2 C1 O o 0

OH "N N N HATU, DIEA, DMF, 30 °C, 70 min N H HO N Ho N HATU, DIEA, DMF, 30 °C, 70 min OH OH

[0770] To a To

[0770] a solution solution of 4-[4-(hydroxymethyl)-1-piperidyl]benzoic of 4-[4-(hydroxymethyl)-1-piperidyl]benzoic acidg,(38 acid (38 g, 161.51 161.51 mmol, mmol,

11 eq) eq) and and 4-(3-amino-2,2,4,4-tetramethyl-cyclobutoxy)-2-chloro-benzonitrile (50.9 4-(3-amino-2,2,4,4-tetramethyl-cyclobutoxy)-2-chloro-benzonitri (50.9 g, 161.51 g, 161.51

mmol,1 1eq, mmol, eq, hydrochloride) hydrochloride) inindimethylformamide dimethylformamide(800(800 mL) mL) was added was added diisopropylethylamine disopropylethylamine

(83.5 (83.5 g, g, 646.04 646.04 mmol, 112mL, mmol, 112 mL,4 eq). 4 eq).The Themixture mixture waswas stirredatat30°C stirred 30°Cforfor10min, 10min, andand then then o-(7 o-(7-

azabenzotriazol-1-yl)-n,n,n',n'-tetramethyluronium hexafluorophosphate azabenzotriazol-1-yl)-n,n,n',n'-tetramethyluronium hexafluorophosphate(64.48 (64.48 g, g,169.59 169.59 mmol, mmol,

1.05 1.05 eq) eq) was added. The was added. Themixture mixturewas wasstirred stirredat at 30 30 °C °C for for 11 hour. hour. LCMS showed LCMS showed the the reaction reaction waswas

completed anddesired completed and desiredMSMS cancan be be detected.TheThe detected. mixture mixture waswas poured poured intointo water water (4L)(4and L) filtered. and filtered. The filter The filter cake cake was was concentrated and and triturated triturated with with methanol (500 mL methanol (500 mLX x2)2)toto give give N-[3-(3- N-[3-(3 chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4- [4-(hydroxymethyl) chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-[4-(hydroxymethyl) -1- -1

piperidyl]benzamide(72 piperidyl]benzamide (72g,g,137.89 137.89mmol, mmol,85%85% yield, yield, 95%95% purity) purity) as aaswhite a white solid. solid.

288

[0771] LCMS:

[0771] LCMS: MS MS (ESI) (ESI) m/z:496.1 m/z: 496.1 [M

[M +1]+ +1]

[0772] ¹H 1H

[0772] NMR: NMR: (400MHz, (400MHz, DMSO-d) DMSO-d) : 7.90 (d, J=8.8 6: 7.90 (d, Hz, 1H), J=8.8 Hz, 7.73 1H), (d, 7.73 J=8.8 Hz, Hz, (d, J=8.8 2H), 2H), 7.48 (d, J=9.2 7.48 (d, J=9.2Hz, Hz, 1H), 1H), 7.207.20 (d, J=2.4 (d, J=2.4 Hz, 7.00 Hz, 1H), 1H),(dd, 7.00 (dd, 8.8 J=2.4, J=2.4, 8.8 Hz, Hz, 1H), 6.95 1H), 6.95Hz, (d, J=8.8 (d, J=8.8 Hz, 2H), 4.48 2H), 4.48(t,(t, J=5.2 J=5.2Hz,Hz, 1H), 1H), 4.314.31 (s, 1H), (s, 1H), 4.05 4.05 (d, J=9.2 (d, J=9.2 Hz, Hz, 1H), 1H), 3.86 (d, 3.86 (d,Hz, J=12.8 J=12.8 Hz,(t, 2H), 3.27 2H), 3.27 (t, J=5.6Hz, J=5.6 Hz,2H), 2H), 2.80 2.80 - 2.70 - 2.70 (m, 2H), (m, 2H), 1.73J=11.2 1.73 (d, (d, J=11.2 Hz, 2H),Hz, 1.632H), 1.63 - 1.52 (m, -1H), 1.521.27 1H), (m,- 1.15 (m, 1.27 - 1.15(m, 8H), 1.12 8H), 1.12(s, (s, 6H). 6H). 2023248067

[0773] Chemical

[0773] Formula: C ChemicalFormula: 2 8 H 3 4 ClN CHCINO, 3 0 3 , Molecular Molecular Weight: Weight: 496.04. 496.04.

[0774] Total

[0774] Total H count HNMR fromHNMR H countfrom data: 34.34. data:

[0775] Step Step

[0775] 4: Synthesis 4: Synthesis of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4- of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4- tetramethyl tetramethyl-

cyclobutyl]-4-(4-formyl-1-piperidyl)benzamide cyclobutyl]-4-(4-formyl-1-piperidyl)benzamide

CI CI 0 CI CI 0 o o O o Dess-Martin Dess-Martin H~"N NH~~~- N N N HN H DCM,3030 N H DCM, OC,2h2h °C, N N N OH OH O

[0776] To a To

[0776] a solution solution ofN-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]- of N-[3-(3-chloro-4-cyano-phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4- 4

[4-(hydroxymethyl)-1-piperidyl]benzamide

[4-(hydroxymethyl)-1-piperidyl]benzamide (65 (65 g, 131.04 g, 131.04 mmol, mmol, 1 eq)1 in eq)dichloromethane in dichloromethane (700 (700 mL)was mL) wasadded added Dess-Martin Dess-Martin reagent reagent (76.70 (76.70 g, 180.83 g, 180.83 mmol, mmol, 1.38 1.38 eq). eq). The The mixture mixture was stirred was stirred at at 30 °C °C for for 2 hours. hours. Thin Thin layer layer chromatography (dichloromethane: chromatography (dichloromethane: methanol=1:1) methanol=1:1) showed showed the the

reaction was reaction completed. The was completed. Thereaction reactionwas wasadjusted adjustedtotopHpH8~98-9 with with saturatedsodium saturated sodium bicarbonate. bicarbonate.

The mixture The mixturewas wasdiluted dilutedwith withwater water(3L) (3 L) and and extractedwith extracted with dichloromethane dichloromethane (1.5(1.5 L x L3). x 3). TheThe

combined organicphase combined organic phasewaswas washed washed withwith saturated saturated brine brine (1.5 (1.5 L xL 2), x 2), driedwith dried withanhydrous anhydrous sodium sulfate,filtered sodium sulfate, filteredandand concentrated concentrated in vacuum. in vacuum. Thewas The residue residue wasbypurified purified by silica gel silica gel

chromatography (dichloromethane: chromatography (dichloromethane: methanol=100:0 methanol=100:0 to 50:1) to 50:1) to give to give N-[3-(3-chloro-4-cyano N-[3-(3-chloro-4-cyano-

phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-(4-formyl- 1-piperidyl)benzamide phenoxy)-2,2,4,4-tetramethyl-cyclobutyl]-4-(4-formyl 1-piperidyl)benzamide (34.6 (34.6 g, 67.94 g, 67.94

mmol,51% mmol, 51% yield,97% yield, 97% purity) purity) as as a whitesolid. a white solid.

[0777] ¹H 1H

[0777] NMR: NMR: (400MHz, (400MHz, DMSO-d) DMSO-d) 6: 9.63 : 9.63 (s, 1H),(s,7.90 1H), (d, 7.90J=8.8 Hz, Hz, (d, J=8.8 1H),1H), 7.74 (d,(d, 7.74 J=8.8 Hz, J=8.8 Hz, 2H), 2H), 7.49 7.49 (d, (d, J=9.2 Hz, 1H), J=9.2 Hz, 1H), 7.20 7.20 (d, (d, J=2.4 Hz, 1H), J=2.4 Hz, 1H), 7.03 - 6.94 7.03 6.94 (m, (m, 3H),4.32 3H), 4.32 (s,(s,1H), 1H), 4.05 (d, 4.05 (d, J=9.2 Hz, 1H), J=9.2 Hz, 1H), 3.76 (td, (td,J=3.6, J=3.6,12.8 12.8Hz, Hz,2H), 3.01 - 2.92 2H), 3.01 2.92 (m, (m, 2H), 2.62 -- 2.55 2H), 2.62 2.55 (m, 1H), (m, 1H), 2.62 - 2.55 2.62 2.55 (m, (m, 1H), 1.92 J=3.6, 1H), 1.92 (dd, (dd, J=3.6, 12.8 12.8 Hz, 2H),Hz, 2H), 1.62 - 1.48 (m,- 2H), 1.62 1.481.21 (m, (s, 2H), 1.21 6H), 1.12(s, 6H), (s, 6H). 1.12 (s, 6H).

[0778] Chemical

[0778] Formula: C ChemicalFormula: 2 8 H 3 2 ClN CHCINO, 3 0 3 , Molecular Molecular Weight: Weight: 494.02. 494.02.

[0779] Total

[0779] Total H count HNMR fromHNMR H countfrom data: 32.32. data:

289

General Synthetic C-4 C-4 Scheme Oct 2023

[0780] General

[0780] Synthetic Scheme

[0781] Synthesis

[0781] Synthesis of building of building blockblock N-((r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl)-4-(4-(2-oxoethyl)piperidin-1-yl)benzamide tetramethylcyclobutyl)-4-(4-(2-oxoethyl)piperidin-1-yl)benzamide

~.0 O 2023248067 10

NN N ZI H N,\ ON NO

CI CI 0 O O

[0782] Synthetic

[0782] Scheme: Synthetic Scheme: 0 0 o o 0 O HO HO- OBn OBn -~OH OH OBn HO NH NH N H2,Pd/C H2,Pd/C N OBn N N DIEA, DMF, 100 °C MeOH HO F F DIEA, DMF, 100 0C HO HO MeOH HO

OH N N NH 2 Na'-O N NH N H HN N Dess-Martin Dess-Martin N N ~ H IZ N CI 0I - N JT aN O DCM, rt, overnight N NO

, DCM, rt, overnight

HATU,DIEA, HATU, DMF DIEA,DMF CI 0 O o CI CI 000

[0783]

[0783] Step 1: Step 1: Synthesis of benzyl Synthesis of 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate benzyl 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate 0

0 HO HO OBn OBn O - OBn OBn NH N N O_ n DIEA, DMF, 100 °C°C HO F F -C DIEA, DMF, 100 Ho

[0784] Into Into

[0784] a a 100-mL round-bottom round-bottom 100-mL flask, flask, was placed was placed benzyl benzyl (2.3 g, (2.3 4-fluorobenzoate 4-fluorobenzoate 10.0g, 10.0 mmol,1.0 mmol, 1.0equiv), equiv), N,N-dimethylformamide N,N-dimethylformamide (30.0(30.0 mL),mL), 2-(piperidin-4-yl)ethan-1-ol 2-(piperidin-4-yl)ethan-1-ol (1.3(1.3 g, 10.0 g, 10.0

mmol,1.0 mmol, 1.0equiv), equiv), N,N-Diisopropylethylamine N,N-Diisopropylethylamine (3.87 (3.87 g, 29.9 g, 29.9 mmol, mmol, 4.0 4.0 equiv). equiv). The The resulting resulting

solution was stirred for was stirred for12 12hhatat90°C. 90°C.The Theresulting resultingmixture mixturewas was concentrated concentrated under vacuum.The under vacuum. The residue was residue wasapplied applied ontoonto a silica a silica gel gel column column withacetate/petroleum with ethyl ethyl acetate/petroleum ether ether (1/1). This(1/1). This resulted in resulted in2.1 2.1gg(62%) (62%) of of benzyl benzyl 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate as aa yellow 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate as yellowsolid. solid.

[0785] LC-MS

[0785] LC-MS (ES):(ES*): 340.25m/z 340.25m/z [MH],[MH+], tR= 1.20min, tR = 1.20min, (1.90 (1.90 minute minute run). run).

[0786] Step Step

[0786] 2: Synthesis 2: Synthesis of 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoic of 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoic acid acid

290

Oct 2023 0 0 O OBn OH Ne N OBn H2,Pd/C H2,Pd/C Ne N OH

HO.C MeOH MeOH HO-C 2023248067 10 Ho Ho

[0787]

[0787] To To aa solution solution of of benzyl benzyl 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate (500mg, 4-[4-(2-hydroxyethyl)piperidin-1-yl]benzoate (500 mg,1.47 1.47 mmol,1.00 mmol, 1.00equiv) equiv)inin20.0 20.0 mL mLmethyl methyl alcohol alcohol (30.0 (30.0 mL)was mL)was added added Pd/C Pd/C (10%, (10%, 300 300 mg) mg) under under nitrogen atmosphere nitrogen atmosphere ininaa 100.0 100.0 mL mLround round bottom bottom flask.TheThe flask. flask flask was was then then vacuumed vacuumed and and flushed with hydrogen. hydrogen. The Thereaction reactionmixture mixturewas washydrogenated hydrogenated at room at room temperature temperature for for 12 hours 12 hours

under hydrogen under hydrogenatmosphere atmosphere using using a hydrogen a hydrogen balloon, balloon, thenthen filteredthrough filtered through a Celitepad a Celite padandand concentrated under reduced concentrated under reducedpressure. pressure. This Thisresulted resulted in in 300.0m 300.Omg g(82.0%) (82.0%)ofof4-[4-(2- 4-[4-(2 hydroxyethyl)piperidin-1-yl]benzoic acidasasaa yellow hydroxyethyl)piperidin-1-yl]benzoic acid yellowsolid. solid.

[0788] LC-MS

[0788] LC-MS (ES):(ES*): 250.00m/z 250.00m/z [MH],[MH+], tR = 0.74min, tR = 0.74min, (2.00 (2.00 minute minute run). run).

[0789]

[0789] 3: Synthesis Step 3: Step Synthesis of 4-[4-(2-hydroxyethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro of 4-[4-(2-hydroxyethyl)piperidin-1-yll-N-[(1r,3r)-3-(3-chloro- 4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide 4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide

OH N o N IOH NH 2 N H OH OH NH N HN N CI O N CI 0 HO HATU, DIEA, DMF HATU, DIEA, DMF CI O 0 o 0 Ho

[0790] Into Into

[0790] a 100-mL a 100-mL round-bottom flask, flask, round-bottom was placed was placed 4-[4-(2-hydroxyethyl)piperidin-1 4-[4-(2-hydroxyethyl)piperidin-1-

yl]benzoic acid yl]benzoic acid (300.0 (300.0 mg, mg, 1.2 1.2 mmol, mmol,2.0 2.0equiv), equiv), N,N-dimethylformamide N,N-dimethylformamide(10.0(10.0 g, 136.8 g, 136.8 mmol, mmol,

227.0 equiv), 227.0 equiv), N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophospate N,N,N,N-Tetramethyl-O-(7-azabenzotriazol-1-yl)uroniun hexafluorophospate

(686 mg, mg, 1.8 1.8 mmol, mmol,3.0 3.0equiv), equiv),2-chloro-4-[(1r,3r)-3-amino-2,2,4,4- 2-chloro-4-[(1r,3r)-3-amino-2,2,4,4 tetramethylcyclobutoxy]benzonitrile hydrochloride(190.0 tetramethylcyclobutoxy]benzonitrile hydrochloride (190.0mg,mg, 0.60.6 mmol, mmol, 1.0 1.0 equiv), equiv), N,N N,N-

Diisopropylethylamine (466.0mg, Diisopropylethylamine (466.0 mg, 3.63.6 mmol, mmol, 6.06.0 equiv). equiv). TheThe resulting resulting solutionwaswas solution stirredfor stirred for11 h at h at room temperature. The room temperature. Thereaction reaction was wasthen thenquenched quenchedby by thethe additionofof6060mLmL addition of of water. water. TheThe

resulting solution resulting solution was was extracted extracted with with 3x30 mLofofethyl 3x30 mL ethyl acetate acetate and and the the organic organic layers layers combined combined

and dried over and dried anhydroussodium over anhydrous sodium sulfateand sulfate andconcentrated concentratedunder under vacuum. vacuum. The The residue residue was was

applied ontoa asilica applied onto silicagelgelcolumn column with with ethyl ethyl acetate/petroleum acetate/petroleum etherThis ether (1:1). (1:1). This in resulted resulted 250.0 in 250.0 mg(81%) mg (81%)ofof4-[4-(2-hydroxyethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)- 4-[4-(2-hydroxyethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy) 2,2,4,4-tetramethylcyclobutyl]benzamide 2,2,4,4-tetramethylcyclobutyl]benzamide asasa ayellow yellowsolid. solid.

[0791] LC-MS

[0791] LC-MS (ES):(ES*): 510.25m/z 510.25m/z [MH],[MH+], tR = 1.35min, tR = 1.35min, (1.90 (1.90 minute minute run). run).

291

[0792] Step Step

[0792] 4: Synthesis 4: Synthesis of 4-[4-(2-oxoethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4 of 4-[4-(2-oxoethyl)piperidin-1-yl]-N-I(1r,3r)-3-(3-chloro-4-

cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide

OH N OHN O N Dess-Martin N N H 1N __O Dess-Martin HN H IN N HN

DCM, rt, overnight

CI N DCM, rt, overnight NNC CI o O 7CI o 00 0

2023248067

[0793] Into Into

[0793] a 100-mL a 100-mL round-bottom round-bottom flask, flask, was placed was placed 4-[4-(2-hydroxyethyl)piperidin-1-yl] 4-[4-(2-hydroxyethyl)piperidin-1-yl]-

N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide (200.0 (200.0 mg, mg, 0.4 0.4 mmol, 1.0 equiv), mmol, 1.0 equiv), dichloromethane dichloromethane(20.0 (20.0mL), mL),Dess-Martin Dess-Martin (249.0 (249.0 mg, mg, 0.600.60 mmol, mmol, 1.5 1.5

equiv). Theresulting equiv). The resulting solution solution was was stirred stirred for 4for 4 hroom h at at room temperature. temperature. The resulting The resulting solution was solution was

extracted extracted with with of of ethyl ethyl acetate acetateand andthe theorganic organiclayers layerscombined combined and concentrated under and concentrated undervacuum. vacuum. Theresidue The residuewaswas applied applied onto onto a silica a silica gel column gel column with with ethyl ethyl acetate/petroleum acetate/petroleum ether (1:1).ether This (1:1). This resulted in resulted in 80.0 80.0 mg (40%)ofof4-[4-(2-oxoethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4- mg (40%) 4-[4-(2-oxoethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4 cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamidea yellow cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamideas as a yellow solid. solid.

[0794] LC-MS

[0794] LC-MS (ES):(ES*): 508.20m/z 508.20m/z [MH],[MH+], tR = 1.19min, tR = 1.19min, (2.00 (2.00 minute minute run). run).

[0795] Exemplary

[0795] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 51 Compound 51

o N O NH N N O N N HN O N NN N ZI H N N ON N

O CIa ;, CI Ó rac-N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(2-(4-(2 rac-N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetraethylcyclobutyl)-4-(4-(2-(4-(2-

(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-6-yl)piperazin-1 (2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-pyrrolol3,4-c|pyridin-6-yl)piperazin-1-

yl)ethyl)piperidin-1-yl)benzamide yl)ethyl)piperidin-1-yl)benzamide

[0796] Synthetic

[0796] scheme: Synthetic scheme:

292

0 0 0 Oct 2023 o o Boc-N NH N OH TMSCHN N o N MeLiOHH BocDEMN Bc S OH OH TMSCHN MeOH C1 O 0i o C1 CI MeOH 2 CI DIEA, DMF DIEA, DMF NN '4 0 MO/2 N MeOH/HO Boc'BocN o H o Boc O o

O O O

2023248067 10

o NH N o 00o NH H N 0 o NH HN NH N N 0 NNN__ N N OH N H2N N NH O 2 N N OH OH HN2 HN 0 o O N CI HOAc Boc N o ___ Boc' HOAc____________ NaBH(OAc), DCM

o O 0 0 N NH NH N N N oN 0

N N #N HN N

O" CI S O 0

107971

[0797] Step 1: Synthesis Step 1: of 3,4-dimethyl Synthesis of 6-chloropyridine-3,4-dicarboxylate 3,4-dimethyl 6-chloropyridine-3,4-dicarboxylate

O O 0 O N NOH OH TMSCHN 2 TMSCHN N0 N C, CI g- OH MeOH C1 CI MeOH O O 0 O

[0798]

[0798] Into aa 100-mL Into round-bottom 100-mL round-bottom flask,was flask, was placed placed 6-chloropyridine-3,4-dicarboxylic 6-chloropyridine-3,4-dicarboxylic acid acid

(200.0 mg, mg, 1.0 1.0 mmol, mmol,1.0 1.0equiv), equiv), methanol methanol(5.0 (5.0mL), mL), acetonitrile(5.0 acetonitrile (5.0 mL), mL),TMSCHN2 TMSCHN2 (2.0 mL), (2.0 mL),

N,N-Diisopropylethylamine N,N-Diisopropylethylamine (516.0 (516.0 mg,mg, 4.Ommol, 4.0mmol, 4.0 equiv). 4.0 equiv). The The resulting resulting solution solution was was stirred stirred

for for 22 hh at atroom room temperature. temperature. The reaction was The reaction was then then quenched quenchedbybythe theaddition additionofofwater water(30mL). (30mL). Theresulting The resultingsolution solution waswas extracted extracted with acetate with ethyl ethyl acetate (20.0 (20.0 mL mLthe x 3)and x 3)and organicthe organic layers layers combined andconcentrated combined and concentratedunder under vacuum. vacuum. The The residue residue was applied was applied onto onto a silica a silica gel gel column column withwith

ethyl acetate/petroleum ethyl acetate/petroleum ether (1:1). (1:1).The The resulting resultingmixture mixture was was concentrated concentrated under vacuum.This under vacuum. This resulted in resulted in 220 220 mg (96%)ofof3,4-dimethyl mg (96%) 3,4-dimethyl6-chloropyridine-3,4-dicarboxylate 6-chloropyridine-3,4-dicarboxylateas as a yellow a yellow solid. solid.

[0799] LC-MS

[0799] LC-MS (ES):(ES*): 230.10m/z 230.10m/z [MH],[MH+], tR = 1.01min, tR = 1.01min, (1.90 (1.90 minute minute run). run).

[0800] Step Step

[0800] 2: Synthesis 2: Synthesis of 3,4-dimethyl of 3,4-dimethyl 6-[4-[(tert-butoxy)carbonyl]piperazin-1 6-[4-[(tert-butoxy)carbonyl]piperazin-1-

yl]pyridine-3,4-dicarboxylate yl|pyridine-3,4-dicarboxylate

293

Oct 2023

o 0 Boc-N Boc-N NH N N

C1 01 O DIEA, DMF N N N O O CI DIEA, DMF N o O Boc'N Boc N 0 2023248067 10

[0801]

[0801] Into aa 100-mL Into round-bottom 100-mL round-bottom flask,was flask, was placed placed 3,4-dimethyl 3,4-dimethyl 6-chloropyridine-3,4 6-chloropyridine-3,4-

dicarboxylate (200.0 mg, dicarboxylate (200.0 mg, 0.9 0.9 mmol, mmol,1.01.0equiv), equiv),N,N-dimethylformamide N,N-dimethylformamide(5.0 (5.0 mL), mL), tert-butyl tert-butyl

piperazine-1-carboxylate (325.0 mg, piperazine-1-carboxylate (325.0 mg,1.7 1.7mmol, mmol,2.02.0equiv), equiv),N,N-Disopropylethylamine N,N-Diisopropylethylamine (450.0 (450.0

mg, 3.5 mg, 3.5 mmol, mmol,4.0 4.0equiv). equiv). The Theresulting resulting solution solution was was stirred stirred for for 22 hh atat100°C. 100°C. The The reaction reaction was was

then quenched then quenchedbybythe theaddition additionof water(80mL).TheThe ofwater(80mL). resultingsolution resulting solutionwas extractedwith wasextracted withethyl ethyl acetate acetate (30.0 (30.0 mL x3) and mL x3) and the the organic organic layers layers combined combinedand andconcentrated concentratedunder under vacuum. vacuum. The The

residue was residue wasapplied applied ontoonto a silica a silica gel gel column column withacetate/petroleum with ethyl ethyl acetate/petroleum ether ether (1:3). This(1:3). This resulted in resulted in 320.0 320.0 mg (97%)ofof3,4-dimethyl mg (97%) 3,4-dimethyl-[4-[(tert-butoxy)carbonyl]piperazin-1-yI]pyridine- 6-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyridine 3,4-dicarboxylate 3,4-dicarboxylate as as a yellow a yellow solid. solid.

[0802] LC-MS

[0802] LC-MS (ES):(ES*): 380.10m/z 380.10m/z [MH],[MH+], tR = 1.19min, tR = 1.19min, (2.0(2.0 minute minute run). run).

[0803] Step Step

[0803] 3: Synthesis 3: Synthesis of 6-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyridine-3,4 of 6-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyridine-3,4-

dicarboxylic acid dicarboxylic acid

0 0 O O N SLiOH LiOH N OH OH O MeOH/H 20 N OH OH N '1 N MeOH/HO N BocN Boc N O 0 BocN Boc N o 0

[0804]

[0804] Into aa 100-mL Into round-bottom 100-mL round-bottom flask, was flask,was placed placed 3,4-dimethyl 3,4-dimethyl 6-[4-[(tert 6-[4-[(tert-

butoxy)carbonyl]piperazin-1-yl]pyridine-3,4-dicarboxylate (320.0mg, butoxy)carbonyl]piperazin-1-yl]pyridine-3,4-dicarboxylate (320.0 mg, 0.8mmol, 0.8 mmol, 1.0 1.0 equiv), equiv),

methanol(10.0 methanol (10.0 mL), mL),water(5 water(5mL), mL), lithium lithium hydroxide hydroxide (96(96 mg,mg, 4 mmol, 4 mmol, 5 equiv). 5 equiv). The The resulting resulting

solution was stirred for was stirred for5 5h hatat room roomtemperature. temperature. The The resulting resultingmixture mixture was was concentrated under under

vacuum. Thisresulted vacuum. This resulted inin 300.0 300.0 mg mg(101%) (101%)of of 6-[4-[(tert-butoxy)carbonyl]piperazin-1 6-[4-[(tert-butoxy)carbonyl]piperazin-1-

yl]pyridine-3,4-dicarboxylic yl]pyridine-3,4-dicarboxylic acid acid as a as a white white solid.solid.

[0805] LC-MS

[0805] LC-MS (ES):(ES*): 296.20m/z 296.20m/z [MH],[MH+], tR = 0.52min, tR = 0.52min, (1.90 (1.90 minute minute run). run).

[0806] Step Step

[0806] 4: Synthesis 4: Synthesis of 3-[1,3-dioxo-6-(piperazin-1-yl)-1H,2H,3H-pyrrolo[3,4 of 3-[1,3-dioxo-6-(piperazin-1-yl)-1H,2H,3H-pyrrolo|3,4-

c]pyridin-2-yl]piperidine-2,6-dione c|pyridin-2-yl|piperidine-2,6-dione

294

Oct 2023 0 00 o o o NH NH N 0 N N N o O NN OOH H 2N H2N o OH N OH : HN O N NOH HN HOAc HOAc N Boc'N Boc o 2023248067 10

[0807]

[0807] Into aa 100-mL Into round-bottom 100-mL round-bottom flask, was flask,was placed placed 6-[4-[(tert-butoxy)carbonyl]piperazin 6-[4-[(tert-butoxy)carbonyl]piperazin-

1-yl]pyridine-3,4-dicarboxylic acid 1-yl]pyridine-3,4-dicarboxylic acid (300.0 (300.0 mg, 0.8 mmol, mg, 0.8 mmol,1.0 1.0equiv), equiv), acetic acetic acid acid (20.0 (20.0 mL), 3 mL), 3-

aminopiperidine-2,6-dione(218 aminopiperidine-2,6-dione (218mg, mg,1.71.7mmol, mmol, 2.02.0 equiv). equiv). TheThe resulting resulting solutionwaswas solution stirredfor stirred for 2 hh at 2 at 130°C. 130°C. The reaction was The reaction was then then quenched quenchedbybythe theaddition additionofofwater water(30 (30mL). mL).The The resulting resulting

solution was extracted with ethyl was extracted ethyl acetate acetate (30 (30 mL x3)andthe mL x3)and the organic organic layers layers combined combinedand and dried dried

in in an an oven oven under reducedpressure. under reduced pressure. and andconcentrated concentratedunder undervacuum. vacuum.TheThe residue residue waswas applied applied

onto aa silica silicagel gelcolumn column with with dichloromethane/methanol (3:1).This dichloromethane/methanol (3:1). Thisresulted resulted in in 60.0 60.0 mg mg(20%) (20%)ofof 3-[1,3-dioxo-6-(piperazin-1-yl)-1H,2H,3H-pyrrolo[3,4-c]pyridin-2-yl]piperidine-2,6-dione as as 3-[1,3-dioxo-6-(piperazin-1-yl)-1H,2H,3H-pyrrolo[3,4-c]pyridin-2-yl]piperidine-2,6-dione a a yellowsolid. yellow solid.

[0808] LC-MS

[0808] LC-MS (ES):(ES*): 344.20m/z 344.20m/z [MH],[MH+], tR= 0.66min, tR = 0.66min, (1.90 (1.90 minute minute run). run).

[0809] Step Step

[0809] 5: Synthesis 5: Synthesis of 4-[4-(2-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1H,2H,3H of 4-[4-(2-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1H,2H,3H-

pyrrolo[3,4-c]pyridin-6-yl]piperazin-1-yl]ethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4 pyrrolo[3,4-c|pyridin-6-yl]piperazin-1-yllethyl)piperidin-1-yl]-N-|(1r,3r)-3-(3-cblor0-4-

cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide ZI

HN HN N o N NN o o NO N oO N N 00N NN N 0 N N HN N N o o O N NH 1 CI o N N NJ NH NaBH(OAC)HH NaBH(OAc),3, DCM DCM O N

HN HN O N N O CI CI

[0810] Into Into

[0810] a 100-mL a 100-mL round-bottom round-bottom flask, flask, was placed was placed 3-[1,3-dioxo-6-(piperazin-1-yl) 3-[1,3-dioxo-6-(piperazin-1-yl)-

1H,2H,3H-pyrrolo[3,4-c]pyridin-2-yl]piperidine-2,6-dione hydrochloride 1H,2H,3H-pyrrolo[3,4-c]pyridin-2-yl]piperidine-2,6-dione hydrochloride (60.0 (60.0 mg,mg, 0.2mmol, 0.2mmol,

1.0 1.0 equiv), equiv), dichloromethane (10 mL), dichloromethane (10 mL),4-[4-(2-oxoethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4- 4-[4-(2-oxoethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4 cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide (80.0 cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide (80.0 mg, mg, 0.1 mmol, 0.1 mmol, 1.0 equiv), 1.0 equiv),

295

Sodiumtriacetoxyborohydride (110.0mg,mg, triacetoxyborohydride(110.0 3.03.0 equiv).TheThe resulting solutionwas was stirredfor for44 hh at at 2023248067 10 Oct 2023

Sodium equiv). resulting solution stirred

roomtemperature. room temperature.The Thereaction reactionwas thenquenched wasthen quenched by by thethe addition addition of of mL mL 40 40 of water. of water. The The

resulting solution resulting solution was was extracted extracted with with dichloromethane (20mL dichloromethane (20 mLx x3)and 3)and theorganic the organiclayers layers combined andconcentrated combined and concentratedunder under vacuum. vacuum. The The crude crude product product (4.0 (4.0 mL)purified mL) was was purified by Prep by Prep-

HPLCwith HPLC with thefollowing the following conditions:Column, conditions: Column, Sunfire Sunfire PrepPrep C18 C18 OBD Column,, OBD Column,,

1Oum,19*250mm; mobile 19*250mm; mobile phase, phase, Water(0.1% Water(0.1% formic formic acid) acid) and acetonitrile and acetonitrile (30.0% (30.0% acetonitrile acetonitrile

up to up to 52.0% in 88 min); 52.0% in min); Detector, Detector, UV UV254nm. 254nm.5.05.0 mL mL product product was was obtained. obtained. This This resulted resulted in 50.5 in 50.5

mg(38.2%) mg (38.2%)ofof4-[4-(2-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1H,2H,3H-pyrrolo[3,4- 4-[4-(2-[4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-1H,2H,3H-pyrrolo[3,4 c]pyridin-6-yl]piperazin-1-yl]ethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy) c]pyridin-6-yl]piperazin-1-yl]ethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-

2,2,4,4-tetramethylcyclobutyl]benzamideasasa ayellow 2,2,4,4-tetramethylcyclobutyl]benzamide yellowsolid. solid.

[0811] 1H NMR

[0811] 1H(300 NMR (300 MHz, MHz, DMSO-d6) DMSO-d6) 6 11.07 11.07 (s, 1H), 8.57(s, (s,1H), 1H),8.57 (s,(d, 7.87 1H), J =7.87 8.7 (d, Hz,J= 8.7 Hz,

1H), 7.70(d, 1H), 7.70 (d,J J= 8.6Hz, = 8.6 Hz, 2H), 2H), 7.447.44 (d, (d, J = J= 9.1 9.1 1H), (s, Hz, 7.29 Hz, 1H), 7.29 1H),(d,7.17 (s,7.17 1H), J = (d, 2.2 J= 1H),Hz, 1H), Hz, 2.2

7.02-6.87 (m,3H), 7.02-6.87 (m, 3H), 5.07 5.07 (dd,(dd, J= 12.8, J = 12.8, 5.3 1H), 5.3 Hz, 1H),(s, Hz, 4.29 4.29 1H), 1H),(s,4.02 (d,4.02 (d, Hz, J = 9.1 J= 1H), 3.28 1H), 9.1 Hz, (s, 3.28 (s,

5H), 2.59-2.41 5H), 2.59-2.41 (m,(m, 9H), 9H), 2.002.00 (t, J(t, = J= 11.3 11.3 Hz, 1.73 Hz, 1H), 1H),(d, 1.73 J =(d, J=Hz,12.8 12.8 2H),Hz, 1.452H), 1.451.14 (s, 3H), (s, 3H), (d, 1.14 (d, J== 27.2 J 27.2 Hz, 14H). Hz, 14H).

[0812] LC-MS

[0812] LC-MS (ES):(ES*): 835.25m/z 835.25m/z [MH],[MH+], tR = 2.56min, tR = 2.56min, (4.80minute (4.80minute run). run).

[0813] Exemplary

[0813] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 52 Compound 52

N NH N O N N N N O N N o N N N NN ZI NN N H N

CI

(rac)-N-((1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(2-(4-(6 (rac)-N-(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobtyl)-4-(4-(2-(4-(6-

(2,6-dioxopiperidin-3-yl)-5,7-dioxo-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)piperazin (2,6-dioxopiperidin-3-yl)-5,7-dioxo-6,7-dihydro-5H-pyrrolo|3,4-dlpyrimidin-2-y/l)piperazin

1-yl)ethyl)piperidin-1-yl)benzamide 1-yl)ethyl)piperidin-1-yl)benzamide

[0814] Synthetic

[0814] scheme: Synthetic scheme:

296

Oct 2023 NH 0.5eq H NH 2SO4 0.5eq HSO S NH 2 NH NH NH NH N N NH Boc'N Boc DIEA, iPrOH, 100 °C, 24 o DIEA, iPrOH, 100 °C, 24 h NH Boc N 2 0 o EtONa, EtOH, reflux, 2h EtONa, EtOH, reflux, 2h N ' 0 o LiOH LiOH 0 o ~NN 'NN o O 2023248067 10

0 o 0 0 N N 0 EtOH/H 20, EtOH/HO, rt, 8h rt, 8h Boc Boc' N o o 0 2 h2 h DMFDMA, rt, rt, DMFDMA, 0 0 o o 0 o 0 N N o O o0

0 HCN HCI NH 0O H2 N NH 0 o o00 o 00 HN o NH N OH Ac2O N N NH N TFA N N OOHNN N o O OH IN N Pyridine,reflux Pyridine, reflux NN O NN DCM DN N N 0 B'N,_ Boc N 0 Bac' N Boc 0 o HN ) HN, o N_ O OF

NOH cNIN N 00 o o o SHNH N NH N o N N NNH2 N N HN N S N o N CI O NH o HN N N N NH ,DCM, NaBH(OAc) 3DCM, NaBH(OAc), I , N o CI o

[0815] Step 1: Synthesis of tert-butyl 4-carbamimidoylpiperazine-1-carboxylat

NH 0.5eq HSO S NH NH NH N N NH Boc DIEA, iPrOH, 100 °C, 24 h BoBoc N Boc

[0816]

[0816] Into Into aa250-ni round-bottom flask, 250-mL round-bottom flask,wasplacedtert-butyl piperazine-1-carboxylate(10 was placed tert-butyl piperazine-1-carboxylate (10

g, g, 53.69 53.69 mmol, 1.00 equiv), mmol, 1.00 equiv), i-propanol i-propanol (150 (150 mL), (methylsulfanyl)methanimidamide mL), (methylsulfanyl)methanimidamide (7.4 (7.4 g, g,

82.09 mmol, 82.09 mmol,1.00 1.00equiv), DIA equiv), DIEA (25(25 mL,mL, 3.00 3.00 equiv). equiv). TheThe resultingsolutionwasstirredfor24h resulting solution was stirred for 24 h

at at 100°C in an 100°C in an oil oilbath. bath.The Theresulting resultingmixture mixturewas was concentrated concentrated under under vacuum. Theresulting vacuum. The resulting solution wasdiluted solution was diluted with with acetonitrile acetonitrile (150 (150 mL),stirred mL), then then stirred for 30 for 30 min. Themin. The solids solids were were

collected byfiltration. collected by filtration. This Thisresulted resulted in in 11.5 11.5 g (94%) g (94%) of tert-butyl of tert-butyl 4-carbamimidoylpiperazine-1 4-carbamimidoylpiperazine-1-

carboxylate carboxylate asas a white a white solid. solid.

[0817]

[0817] Step 2: Step 2: Synthesis Synthesis of of 1,4-diethyl 1,4-diethyl (2Z)-2-[(dimethylamino)methylidene|-3- (2Z)-2-[(dimethylamino)methylidene]-3 oxobutanedioate oxobutanedioate

297

Oct 2023 0 0 0 0 DMFDMA, DMFDMA, rt,rt,2 2h h O O

N 0 O 0 N O 2023248067 10

[0818] Into Into

[0818] a 250-mL a 250-mL round-bottom round-bottom flask, flask, was placed was placed 1,4-diethyl 1,4-diethyl 2-oxobutanedioate 2-oxobutanedioate (10 g, (10 g, 53.14 mmol,1.00 53.14 mmol, 1.00equiv), equiv), DMFDMA DMFDMA(12.65(12.65 g, 106.30 g, 106.30 mmol, mmol, 2.00 equiv) 2.00 equiv) at The at 0 °C. 0 °C.resulting The resulting solution wasstirred solution was stirredforfor2 h2 at h atroom room temperature. temperature. The residue The residue wasonto was applied applied ontogela silica gel a silica

column with column with ethyl ethyl acetate/petroleum acetate/petroleum ether (7/3). ether (7/3). This resulted This resulted in 2.79 gin(22%) 2.79ofg 1,4-diethyl (22%) of 1,4-diethyl (2Z)-2-[(dimethylamino)methylidene]-3-oxobutanedioate (2Z)-2-[(dimethylamino)methylidene]-3-oxobutanedioate as yellow as yellow oil. oil.

[0819] LC-MS

[0819] LC-MS (ES):(ES*): m/z 243.95 m/z 243.95 [MH],[MH+], tR =0.64min, tR =0.64min, (1.90minute (1.90minute run). run).

[0820] Step Step

[0820] 3: Synthesis 3: Synthesis of 4,5-diethyl of 4,5-diethyl 2-[4-[(tert-butoxy)carbonyl]piperazin-1 -[4-[(tert-butoxy)carbonyl]piperazin-1-

yl]pyrimidine-4,5-dicarboxylate yl|pyrimidine-4,5-dicarboxylate

NH NH 0 00 o EtONa, EtOH, reflux, 2h N NN NH 2 O EtONa, EOH, reflux, 2h N NH o0D N NN o BocBN Boc N N 1Boc'N N 0 O Boc

[0821]

[0821] Into Into aa 250-mL round-bottom 250-mL round-bottom flask, was flask,was placed placed tert-butyl 4-carbamimidoylpiperazine tert-butyl4-carbamimidoylpiperazine- 1-carboxylate (1.0 g, 1-carboxylate (1.0 g, 4.38 4.38 mmol, 1.00 equiv), mmol, 1.00 equiv), ethanol ethanol (20 (20 mL), 1,4-diethyl (2Z)-2 mL), 1,4-diethyl (2Z)-2-

[(dimethylamino)methylidene]-3-oxobutanedioate

[(dimethylamino)methylidene]-3-oxobutanedioate (1.065 (1.065 g, 4.38 g, 4.38 mmol, mmol, 1.00 1.00 equiv), equiv), EtONaEtONa (596 (596 mg,8.76 mg, 8.76mmol, mmol, 1.00 1.00 equiv). equiv). The resulting The resulting solutionsolution wasfor was stirred stirred 2 h atfor 2 hinatan75°C 75°C in anThe oil bath. oil bath. The mixture was resulting mixture resulting concentrated under was concentrated undervacuum. vacuum.TheThe resulting resulting solutionwas solution was extracted extracted with with

ethyl ethyl acetate acetate (100 (100 mL) and the mL) and the organic organic layers layers combined. combined.The Theresulting resulting mixture mixturewas waswashed washed with with

brine (100 mL). brine mL). The Themixture mixturewas wasdried driedover overanhydrous anhydrous sodium sodium sulfate. sulfate. TheThe residue residue was was applied applied

onto onto aasilica silica gel gel column column with with ethyl ethyl acetate/petroleum acetate/petroleum ether This ether (1/5). (1/5).resulted This resulted in 873.0 mg in 873.0 mg

(49%) of4,5-diethyl (49%) of 4,5-diethyl 2-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyrimidine-4,5-dicarboxylate 2-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyrimidine-4,5-dicarboxylate asas

light light yellow oil. yellow oil.

[0822] LC-MS

[0822] LC-MS (ES):(ES*): m/z 409.20 m/z 409.20 [MH+],

[MH], tR = tR = 1.19min, 1.19min, (1.90minute (1.90minute run). run).

[0823] Step Step

[0823] 4: Synthesis 4: Synthesis of2-[4-[(tert-butoxy)carbonyl]piperazin-1-yl]pyrimidine-4,5 of2-[4-[(tert-butoxy)carbonyl|piperazin-1-yl|pyrimidine-4,5-

dicarboxylic acid dicarboxylic acid

298

Oct 2023 0 O 0 O N N' 0 ,LiOH OH LiOH N OH N N O EtOH/HO,2 0, OEtOH/H rt,rt,88 hh N N OH OH N N N N Boc'N Boc N o 0 Boc'N Boc N O 0 2023248067 10

[08241

[0824] Into aa 100-mL Into round-bottom 100-mL round-bottom flask, was flask,was placed placed 4,5-diethyl 2-[4-[(tert 4,5-diethyl2-[4-[(tert- butoxy)carbonyl]piperazin-1-yl]pyrimidine-4,5-dicarboxylate(873.0 butoxy)carbonyl|piperazin-1-yl]pyrimidine-4,5-dicarboxylate (873.0 mg,mg, 2.14 2.14 mmol, mmol, 1.00 1.00 equiv), equiv),

ethanol/water(5/2) (14 ethanol/water(5/2) mL), lithium (14 mL), lithium hydroxide hydroxide(256.7 (256.7mg, mg,10.72 10.72mmol, mmol, 5.00 5.00 equiv). equiv). TheThe

solutionwaswas resulting solution resulting stirred stirred forfor 8 h 8at h room at room temperature. temperature. The resulting The resulting mixture mixture was was concentrated under concentrated under vacuum. vacuum. This resulted This resulted in 1.02 in 1.02 g of g (crude) (crude) of 2-[4-[(tert 2-[4-[(tert-

butoxy)carbonyl]piperazin-1-yl]pyrimidine-4,5-dicarboxylic butoxy)carbonyl]piperazin-1-yl|pyrimidine-4,5-dicarboxyli acidacid aswhite as a a white solid. solid.

[0825] LC-MS

[0825] LC-MS (ES):(ES*): m/z 352.45 m/z 352.45 [MH+],

[MH], tR = tR = 0.73min, 0.73min, (1.90minute (1.90minute run). run).

[0826] Step Step

[0826] 5: Synthesis 5: Synthesis of tert-butyl of tert-butyl 4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo 4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-

5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2-y]piperazine-1-carboxylate 5H,6H,7H-pyrrolol3,4-d|pyrimidin-2-yllpiperazine-1-carboxylate 0 HCI O HCI NH o H 2NtNH 0 o0 H2N O OH Ac O ON NH N OH 2 N N 0 N1N OH - AcO r N N o OH N N N N Pyridine, reflux Pyridine, reflux B N N Boc'N, o 0 Boc NN Boc 0 o Boc

[0827]

[0827] Into aa 100-mL Into round-bottom 100-mL round-bottom flask, was flask,was placed placed 2-[4-[(tert-butoxy)carbonyl]piperazin 2-[4-[(tert-butoxy)carbonyl]piperazin-

1-yl]pyrimidine-4,5-dicarboxylic acid 1-yl]pyrinmidine-4,5-dicarboxylic acid (735.0 (735.0 mg, mg, 2.09 mmol, 1.00equiv). mmol, 1.00 equiv). This This was wasfollowed followedbyby the addition the additionofofacetic aceticanhydride anhydride (10 (10 mL), mL), after after stirred stirred 2h at 2h 130 at °C,130 °C, concentrated concentrated under under vacuum. vacuum. To this To this was addedpyridine was added pyridine (10 (10 mL), mL),3-aminopiperidine-2,6-dione 3-aminopiperidine-2,6-dione hydrochloride hydrochloride (445.0 (445.0 mg, mg, 2.702.70

mmol,1.30 mmol, 1.30 equiv). equiv). The The resulting resulting solution solution was stirred was stirred overnight overnight at an at 120°C in 120°C in anThe oil bath. oil bath. The resulting mixture resulting mixture was concentrated under was concentrated undervacuum. vacuum.TheThe resultingsolution resulting solutionwas was dilutedwith diluted with dichloromethane (100mL). dichloromethane (100 mL). TheThe solids solids were were filteredout. filtered out. The Theresidue residuewas wasapplied appliedonto ontoa asilica silica gel column gel column with with ethyl ethyl acetate/petroleum acetate/petroleum ether This ether (7/3). (7/3).resulted This resulted in 243.0 in mg 243.0 mgtert- (26%) of (26%) of tert butyl 4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2 butyl 4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-djpyrimidin-2-

yl]piperazine-1-carboxylate as yl|piperazine-1-carboxylate as brown brownoil. oil.

[0828] LC-MS

[0828] LC-MS (ES):(ES*): m/z 467.10 m/z 467.10 [M Na*],

[M Na], = 1.10min, tr = tR1.10min, (2.00minute (2.00minute run). run).

[0829] Step Step

[0829] 6: Synthesis 6: Synthesis of3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H-pyrrolo[3,4 of3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H-pyrrolo|3,4-

d]pyrimidin-6-yl]piperidine-2,6-dione d|pyrimidin-6-yl|piperidine-2,6-dione

299

Oct 2023 0 0 O 0 0 O o N NH NH N O TFA N N O NN N N TDCM N DCM NN N N Boc'N Boc N o 0 HN HN 0 O 2023248067 10

[0830]

[0830] Into aa 50-mL Into round-bottom 50-mL round-bottom flask, wasplaced flask,was 4-[6-(2,6-dioxopiperidin-3 tert-butyl4-[6-(2,6-dioxopiperidin-3- placedtert-butyl yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2-yl]piperazine-1-carboxylate yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2-yllpiperazine-1-carboxylate (243.0 (243.0 mg, 0.55 mg, 0.55

mmol,1.00 mmol, 1.00equiv), equiv), dichloromethane dichloromethane(5.0 (5.0mL), mL), trifluoroaceticacid trifluoroacetic acid(2.0 (2.0 mL). mL).The Theresulting resulting solution solution was stirred for was stirred for2 2h hatat room roomtemperature. temperature. The The resulting resultingmixture mixture was was concentrated under concentrated under

vacuum. Thisresulted vacuum. This resulted inin 320.0 320.0 mg mg(crude) (crude)ofof3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H- 3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H pyrrolo[3,4-d]pyrimidin-6-yl]piperidine-2,6-dione asasbrown pyrrolo[3,4-d]pyrimidin-6-yl]piperidine-2,6-dione brownoil. oil.

[0831] LC-MS

[0831] LC-MS (ES):(ES*): m/z 345.25 m/z 345.25 [MH+],

[MH], tR = tR = 0.61min, 0.61min, (1.90minute (1.90minute run). run).

[0832] Step Step

[0832] 7: Synthesis 7: Synthesis of 4-[4-(2-[4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H of 4-[4-(2-|4-|6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H-

pyrrolo[3,4-d]pyrimidin-2-yl]piperazin-1-yl]ethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4 pyrrolo[3,4-d|pyrimidin-2-yl|piperazin-1-yl|ethyl)piperidin-1-yl|-N-[(1r,3r)-3-(3-chloro-4-

cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide cyanophenoxy)-2,2,4,4-tetramethylcyclobutyllbenzamide HN H HN o o o N N NN~ Ny: N N 0 N HN --- N 0 N N N O 0 N0 N N 0__NH_ o o CI ____CD N (~I~j~HNaBH(OAC) NH ,DCM, NaBH(OAc), 3DCM, 0 O N~ O N N

H HN N O NZ \ NE 0 CI CI

[0833] Into Into

[0833] a 100-mL a 100-mL round-bottom round-bottom flask, flask, was placed was placed 4-[4-(2-oxoethyl)piperidin-1-yl]-N 4-[4-(2-oxoethyl)piperidin-1-yl]-N-

[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide(90 (90

[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]benzamide mg, mg, 0.180.18

mmol,1.00 mmol, 1.00equiv), equiv), dichloromethane dichloromethane(10(10mL), mL), 3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H 3-[5,7-dioxo-2-(piperazin-1-yl)-5H,6H,7H-

pyrrolo[3,4-d]pyrimidin-6-yl]piperidine-2,6-dione (61.24 pyrrolo[3,4-d]pyrimidin-6-yl|piperidine-2,6-dione (61.24mg, mg,0.18 0.18mmol, mmol, 1.00 1.00 equiv). equiv). This This waswas

followed followed byby thethe addition addition of DIEA of DIEA (0.5after (0.5 mL), mL),stirred after stirred at 30 °Catfor 301h. °C To forthis 1h. was To added this was added NaBH(OAc)3 NaBH(OAc) (122.89 (122.89 mg, mmol, mg, 0.58 0.58 mmol, 3.00 equiv). 3.00 equiv). The resulting The resulting solution solution was stirred was stirred for 5for 5 h h at at 30 °Cin an 30 °Cin an oil oil bath. bath.The The resulting resultingsolution solutionwas wasextracted extractedwith withdichloromethane (150 mL) dichloromethane (150 mL)and andthe the organic layers combined. organic layers Theresulting combined. The resulting mixture mixturewas waswashed washed with with brine brine (50(50 mL). mL). The The mixture mixture

was dried over was dried over anhydrous anhydroussodium sodium sulfateand sulfate andconcentrated concentrated under under vacuum. vacuum. The The crudecrude product product

300 was purified purified by Prep-HPLC with thethe following conditions: Column, XBridge Prep Prep C18 OBD 2023248067 10 Oct 2023 was by Prep-HPLC with following conditions: Column, XBridge C18 OBD

Column, 5um,19*150 Column, 5um, 19*150 mm; mm; mobile mobile phase, phase, waterwater (1Ommol/L (10mmol/L bicarbonate bicarbonate amine) amine) and and acetonitrile acetonitrile

(30.0% acetonitrile up (30.0% acetonitrile up to to 51.0% in 88 min); 51.0% in min); Detector, Detector, UV 254nm. UV 254nm. This This resultedinin5050mgmg resulted (34%) (34%) of of

4-[4-(2-[4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2 4-[4-(2-[4-[6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-5H,6H,7H-pyrrolo[3,4-d]pyrimidin-2-

yl]piperazin-1-yl]ethyl)piperidin-1-yl]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4 yl]piperazin-1-yl]ethyl)piperidin-1-yI]-N-[(1r,3r)-3-(3-chloro-4-cyanophenoxy)-2,2,4,4-

tetramethylcyclobutyl]benzamide tetramethylcyclobutyl]benzamide as as a ayellow yellow solid. solid.

[0834]

[0834] ¹H 'H NMR NMR (400 MHz, (400 d6-DMSO): MHz, 811.12 611.12 d6-DMSO): (s, 1H),(s,8.90 1H),(s, 1H), 8.90 (s, 7.91-7.89 (d, J=8.4Hz, 1H), 7.91-7.89 (d, J=8.4Hz, 1H), 1H), 7.74-7.72 (d, J=7.6Hz, 7.74-7.72 (d, 2H), 7.49-7.47 J=7.6Hz, 2H), 7.49-7.47 (d, (d, J=8.8Hz, 1H),7.20 J=8.8Hz, 1H), 7.20(s, (s, 1H), 6.99-6.94 (m, 1H), 6.99-6.94 (m, 3H), 3H), 5.16-5.13 5.16-5.13 (m, 1H), 4.32 (m, 1H), (s, 1H), 4.32 (s, 1H), 4.06-3.83 4.06-3.83 (m, (m, 7H), 7H), 2.88-2.57 (m, 5H), 2.88-2.57 (m, 5H), 2.39-2.33 2.39-2.33 (m, (m, 2H), 2H), 2.07- 2.07 2.01 (m, 2.01 1H), 1.78-1.75 (m, 1H), 1.78-1.75 (m, (m, 2H), 2H), 1.54-1.35 1.54-1.35 (m, (m, 3H), 3H),1.21 1.21 (m, (m,8H), 8H),1.12 1.12(s, (s, 6H); 6H); LC-MS LC-MS (ES*): (ES):

m/z 836.45/838.45 m/z 836.45/838.45[MH],

[MH+], = 2.17min, tR =tR2.17min, (2.95minute (2.95minute run). run).

[0835] Chemical

[0835] Chemicalformula: formula: CCHCINO 4 4 H5 0 ClN 90 6 [835.36/837.36].

[835.36/837.36].

[0836]

[0836] Total H Total H count HNMR fromHNMR count from data: 50. 50. data:

[0837]

[0837] D. Exemplary Synthetic Schemes Exemplary Synthetic for Exemplary Schemes for BRaf ExemplaryBRaf Targeting Targeting Moiety Moiety Based Based

Compounds Compounds

[0838]

[0838] General General Synthetic Approach Synthetic Approach The synthetic

[0839] The synthetic

[0839] realization and and realization optimization optimization of the of the bifunctional bifunctional molecules molecules as described as described

herein may herein beapproached may be approachedin ina astep-wise step-wiseorormodular modular fashion.For fashion. Forexample, example, identificationofof identification

compounds thatbind compounds that bindtotothe thetarget target molecules moleculescan caninvolve involvehigh highorormedium medium throughput throughput screening screening

campaigns if no campaigns if no suitable suitable ligands ligands are immediately are immediately available. available. It is notItunusual is not unusual forligands for initial initialtoligands to require iterative require iterative design designandand optimization optimization cycles cycles to improve to improve suboptimal suboptimal aspects as aspects as by identified identified by data from suitable in from suitable in vitro vitroand andpharmacological and/or ADMET pharmacological and/or assays. ADMET assays. PartPart of the of the

optimization/SAR campaign optimization/SAR campaign would would beprobe be to to probe positions positions of the of the ligand ligand that that arearetolerant tolerantofof substitution andthat substitution and thatmight might be suitable be suitable places places on which on which to the to attach attach thechemistry linker linker chemistry previously previously

referred toto herein. referred herein.Where Where crystallographic crystallographic or NMRor NMR structural structural data are available, data are available, these these can be can be usedtotofocus used focussuch such a synthetic a synthetic effort. effort.

[0840] In a Invery

[0840] a very analogous analogous waycan way one oneidentify can identify and optimize and optimize ligands ligands for anfor E3an E3 Ligase, Ligase, i.e. i.e. ULMs/ILMs/VLMs/CLMs/ILMs. ULMs/ILMs/VLMs/CLMs/ILMs. With

[0841] With

[0841] PTMs PTMs and ULMs (e.g.(e.g. and ULMs ILMs,ILMs, VLMs, VLMs, CLMs,CLMs, ILMs) ILMs) and/orand/or in hand, in hand, one skilled one skilled

in the the art artcan canuse useknown known synthetic synthetic methods for their methods for their combination with or combination with or without withoutaa linker linker moiety. moiety.

Linker moieties Linker moieties can can be be synthesized synthesized with withaa range range of ofcompositions, compositions,lengths lengthsand andflexibility flexibility and and

301

Oct 2023 functionalized functionalized such that the such that the PTM andULM PTM and ULM groups groups can can be attached be attached sequentially sequentially to distal to distal ends ends of of

the linker. Thus the linker. libraryofof Thusa alibrary bifunctional bifunctional molecules molecules can becan be realized realized and profiled and profiled in and in in vitro in vitro in and in vivo pharmacological andADMET/PK pharmacological and ADMET/PK studies. studies. As the As with withPTM theand PTMULM and ULMthe groups, groups, final the final

bifunctional molecules bifunctional molecules can can be subject be subject to iterative to iterative designdesign and optimization and optimization cycles in cycles order toin order to 2023248067 10

identify molecules identify molecules with with desirable desirable properties. properties.

[0842] In some

[0842] In some instances, instances, protecting protecting group group strategies strategies and/or and/or functional functional group group

interconversions (FGIs) interconversions (FGIs) may may be required be required to facilitate to facilitate the preparation the preparation of thematerials. of the desired desired materials. Such chemical Such chemicalprocesses processesare arewell wellknown knownto to thesynthetic the syntheticorganic organicchemist chemistandand many many of these of these maymay

be found be found in in texts texts such such as as "Greene's "Greene's Protective Protective Groups in Organic Groups in Organic Synthesis" Synthesis"Peter Peter G. G. M. M.Wuts Wuts and TheodoraW.W.Greene and Theodora Greene (Wiley), (Wiley), andand "Organic "Organic Synthesis: Synthesis: The The Disconnection Disconnection Approach" Approach" Stuart Stuart

Warren andPaul Warren and PaulWyatt Wyatt (Wiley). (Wiley).

General

[0843] General

[0843] Synthetic Scheme SyntheticScheme D-1 D-1

Br Br- Br Br.

NH + N-Ar-L-3-PG NNH N + M'-Ar-L--CN-PG M'-Ar-L -fJJPG N N N NN N Il' / XVII XVII XVI XVI II'

M-Ar' M-Ar' Ar' Ar' Ar'

XVIII XVIII '- NN-Ar-L-J NHH ~N-Ar-L N-Ar-L -PG N-PG N N ' NN N N N N XIX XIX X XX

O o NH o O N o NH YN ~ O Ar' Ar' NN 0 o X Y VII VII - NN-Ar-L -A -L N NNN N XXI XXI

[0844] A compound

[0844] A compound of formula of formula XVI mayXVI may bewith be reacted reacted with a II' a reagent reagent II' (commercially (commercially

available available ororreadily readilyprepared prepared using using standard standard reaction reaction techniques techniques known to known to one one skilled skilled in the art) in the art)

under Chan-Lam under Chan-Lam cross-coupling cross-coupling conditions, conditions, e.g.copper e.g. copper (II)acetate, (II) acetate, pyridine pyridine or or diethylamine diethylamine or or triethylamine, triethylamine, 100 °C, to to produce a compound produce a compound of of formula formula XVII. XVII. M' represents M' represents a boronic a boronic acidacid or or

302 boronicester; represents ester;ArArrepresents an aromatic or heteroaromatic ring Lsystem; L represents an optional 2023248067 10 Oct 2023 boronic an aromatic or heteroaromatic ring system; represents an optional linker, linker, represents represents aa primary primary or or secondary amine, optionally secondary amine, optionally cyclized into aa 44 to cyclized into to8 8membered membered heterocyclicring, heterocyclic ring,wherein wherein PG represents PG represents a suitable a suitable protecting protecting group, including group, including but not but not limited to limited to t-butoxycarbonyl or benzyl. t-butoxycarbonyl or benzyl. Compounds Compounds of formula of formula XVII XVII may may be maybe bemay be reacted reacted with a with a reagent reagent

XVIII underpalladium-catalyzed XVIII under palladium-catalyzedcross-coupling cross-coupling conditions,e.g. conditions, e.g.[1,1'-

[1,1' bis(diphenylphosphino)ferrocene]dichloropalladium, tri-tert-butylphosphinetetrafluoroborate, bis(diphenylphosphino)ferrocene]dichloropalladium, tri-tert-butylphosphine tetrafluoroborate, cesium fluoride, 1,4-dioxane, cesium fluoride, 1,4-dioxane, 90 °C, to 90 °C, to produce produce aa compound compound of of formula formula XIX. XIX. M represents M represents a a functional functional group capable of group capable of undergoing undergoingpalladium-catalyzed palladium-catalyzedtransmetallation, transmetallation,e.g. e.g. aa boronic boronic acid, acid, boronicester, boronic ester,orortrialkylstannane trialkylstannaneand and Ar' represents Ar' represents an aromatic an aromatic or heteroaromatic or heteroaromatic ring system ring system with optional with optional substituents. substituents. A compound A compound of of formula formula XIXXIX may may then then be converted be converted to a to a compound compound

of of formula XXbybytreatment formula XX treatmentwith witha areagent reagentsuitable suitablefor for the the removal ofPG, removal of PG,e.g. e.g. hydrogen hydrogen chloride chloride in in 1,4-dioxane 1,4-dioxane or or methanol whenPGPG methanol when is is t-butyl. A Acompound t-butyl. compound of formula of formula XXalso XX may may also be reacted be reacted with a compound with a compound ofof formula formula VII VII to to provide provide compounds compounds of formula of formula XXI, XXI, wherein wherein X is X is aa suitable suitable leaving leavinggroup group such such as fluorine as fluorine or chlorine, or chlorine, Y isthe Y is C=O, C=O, the aromatic aromatic ring ring of VII may of VII may

havefurther have furtheroptional optional substituents, substituents, and and reaction reaction conditions conditions arefor are those those for a nucleophilic a nucleophilic aromatic aromatic substitution, substitution,e.g. e.g.triethylamine, DMSO, triethylamine, 80 °C. DMSO, 80 °C. InIn cases cases where wherethe thegroup groupAr' Ar'contains containsoptional optional substituents, e.g. aaketone, substituents, e.g. ketone,these thesemaymay undergo undergo further further functionalization, functionalization, e.g. by treatment e.g. by treatment with with hydroxylaminehydrochloride hydroxylamine hydrochloride andand pyridine pyridine at at room room temperature, temperature, to provide to provide further further compounds compounds

of of formula XXI. formula XXI.

[0845]

[0845] General General Synthetic Scheme Synthetic Scheme D-2 D-2

Br Br Br Br-

N-Ar-L-4J.J-PG N-PG N-Ar-L-5H N-Ar-L NH N N N N N N XXII XXII XVII XVII

O 0 o XVIII XVIII NH Br- N-A-_P Y N 0 o xxi XXI V1Br VII VII ,,N-Ar-L N-Ar-L Y N N N N N /

XXIII XXIII

303 be may be converted to a compound of 2023248067 10 2023

Alternatively,

[0846] Alternatively,

[0846] a compound a compound of formula of formula XVII XVII may converted to a compound of

formula XXIIbybyusing formula XXII conditionsanalogous usingconditions to to analogous thoseforforthe those conversionofofXIX theconversion XIX XXXX to to in in

Oct Scheme5.5.A A Scheme compound compound of formula of formula XXII XXII maybethen may then be treated treated with awith a compound compound of formula of formula VII VII as defined defined in in Scheme Scheme 5 5to to produce produceaacompound compoundof of formula formula XXIII. XXIII. The compound The compound of formula of formula

XXIII maythen XXIII may thenbebetreated treatedwith withaareagent reagent XVIII XVIIIasasdefined definedininScheme Scheme 5 toproduce 5 to produce a compound a compound

of formulaXXI. of formula XXI. In cases In cases wherewhere the Ar' the group group Ar' optional contains containssubstituents, optional substituents, e.g.these e.g. a ketone, a ketone, these mayundergo may undergofurther furtherfunctionalization, functionalization, e.g. e.g. by by treatment treatment with with hydroxylamine hydrochlorideandand hydroxylamine hydrochloride

pyridine pyridine at at room temperature, to room temperature, to provide further compounds provide further compounds ofof formula formula XXI. XXI.

[0847] Exemplary

[0847] Exemplary Synthesis Exemplary Compound of Exemplary Synthesis of 42: Compound 42:

(E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-(hydroxyimino)-2,3-dihydro-1H-inden

[0848] (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-(hydroxyimino)-2,3-dihydro-lH-inden-

[0848]

5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)-4-methylisoindoline-1,3-dione

HO'N HO-N

- - 0 NN N NF N N NO N N NH NH N N 0 O

[0849] Step Step

[0849] A: 2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro-1H A: 2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro-1H-

inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione

0 o

O N N ~NN~N\/ N 0 NN N N N N OO NH NH N N 0 O

[0850] To a To

[0850] a solution solution of 4-chloro-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-oxo-2,3-dihydro of 4-chloro-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-oxo-2,3-dihydro-

1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione 1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione

(100 mg, mg, 0.14 0.14 mmol) mmol) inl,4-dioxane in1,4-dioxane 10 10 mL mL and and H2O H20 1 mL 1were mLadded weremethylboronic added methylboronic acid (33.6 acid (33.6

mg, 0.56 mg, 0.56 mmol), mmol),Pd(aMPhos)Cl Pd(aMPhos)C12 (9.9 (9.9 mg, 0.014 mg, 0.014 mmol), mmol), and and CsF CsF (85.12 (85.12 mg, mg, 0.56 0.56 The mmol). mmol). The resulting solution resulting solutionwaswas irradiated irradiated at °C at 90 90 with °C with MW forMW 2 h. for 2 h. After Aftertocooling cooling to rt, rt, it was it was diluted diluted with EA with EA(50 (50mL), mL),and andthe themixture mixturewaswas washed washed withwith brine brine (3 x(3 20 x 20 mL). mL). The The organic organic phasephase was was dried over Na2SO4, filteredand NaSO, filtered andconcentrated concentrated under under reduced reduced pressure. pressure. TheThe residue residue waswas purified purified

by prep-TLC by prep-TLCtotoafford afford2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro-1H- 2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro-1H

304

Oct 2023 inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione (70 (/0

mg, 72.1% mg, 72.1% yield). yield). LCMS (ES*): m/z LCMS (ES): m/z 706.3 706.3 [M+H]*.

[M+H].

[0851] Step Step

[0851] B: (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-(hydroxyimino)-2,3-dihydro B: (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4-(1-(hydroxyimino)-2,3-dihydro-

1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)-4-methylisoindoline 1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)-4-methylisoindoline- 2023248067 10

1,3-dione 1,3-dione

HO-N HO~N

N N / NN O N N N O N N N NH NH N-' 0 O N O

[0852] To a To

[0852] a solution solution of 2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro of 2-(2,6-dioxopiperidin-3-yl)-4-methyl-5-(4-(4-(4-(1-oxo-2,3-dihydro-

1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione 1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-yl)phenyl)piperazin-1-yl)isoindoline-1,3-dione

(70 (70 mg, 0.10 mmol) mg, 0.10 mmol)ininacetonitrile acetonitrile 33 mL mLand andpyridine pyridine3 3mLmL was was added added hydroxylamine hydroxylamine

hydrochloride (69.5 mg, hydrochloride (69.5 mg,1.0 1.0 mmol). mmol).The The mixture mixture waswas stirredatat4040°C°Cforfor2020min. stirred min.Then Then it itwas was diluted diluted with with DCM (20mL), DCM (20 mL), andand thethe mixture mixture waswas washed washed with with brinebrine (10 mL). (10 mL). The organic The organic phase phase

was concentrated was concentratedand andpurified purified by byprep-TLC prep-TLCto to afford(E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4- afford (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(4 (1-(hydroxyimino)-2,3-dihydro- 1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1 (1-(hydroxyimino)-2,3-dihydro-1H-inden-5-yl)-3-(pyridin-4-yl)-1H-pyrazol-1-

yl)phenyl)piperazin-1-yl)-4-methylisoindoline-1,3-dione (19.6 yl)phenyl)piperazin-1-yl)-4-methylisoindoline-1,3-dione (19.6mg, mg,27.8% 27.8% yield) yield) as as yellow yellow solid. solid.

H NMR ¹H NMR (400 (400 MHz, MHz, DMSO-d): DMSO-d): 11.09 (s, 1H), (s, 6 11.09 10.89 1H),(s, 1H),(s, 10.89 8.72 1H),(s,8.72 1H), (s,8.58-8.57 (m, 2H), 1H), 8.58-8.57 (m, 2H), 7.83 (d, JJ= 7.83 (d, 8.0 Hz, = 8.0 Hz,2H), 2H), 7.73 7.73 (d, (d, J = J= 7.6 7.6 1H), 1H), Hz, Hz, 7.56J (d, 7.56 (d, J=Hz,7.61H), = 7.6 1H), 7.50-7.41 Hz,7.50-7.41 (m, 4H), (m, 4H),

7.23-7.17 (m, 3H), 7.23-7.17 (m, 3H), 5.13-5.09 5.13-5.09 (m, 1H), 3.61-3.42 (m, 1H), 3.61-3.42 (m, (m,8H), 8H),3.04-2.97 3.04-2.97(m, (m,2H), 2H),2.93-2.82 2.93-2.82(m, (m,3H), 3H), 2.62-2.56 (m, 2.62-2.56 (m, 5H), 5H), 2.08-2.00 2.08-2.00 (m, 1H); LCMS (m, 1H); LCMS (ES*): (ES): m/z 721.3 m/z 721.3 [M+H]*.

[M+H].

Exemplary

[0853] Exemplary

[0853] Compound Compound 41 may may 41 be prepared by aby be prepared a procedure procedure analogous to tothat analogous that described for described for Examplary Compound Examplary Compound 42. 42.

[0854] E. E.

[0854] Exemplary Exemplary Synthetic Synthetic Schemes for for Schemes Exemplary BRD4 BRD4 Exemplary Binding MoietyMoiety Binding Based Based Compounds Compounds

[0855] Exemplar

[0855] Exemplar Synthesis Synthesis of Exemplary of Exemplary Compound Compound 45: 2-((S)-4-(4-chlorophenyl) 45: 2-((S)-4-(4-chlorophenyl)-

2,3,9-trimethyl-6H-thieno[3,2-f11,2,4]triazolo[4,3-a1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-((3 2,3,9-trimethyl-6H-thieno|3,2-f]|1,24]triazoloj4,3-a|l14|diazepin-6-yl)-N-(4-(2-(2-(2-(2-(3-

(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7 (2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-34-dihydroquinazolin-7-

yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide

305

Oct 2023 O NH HCI NH2HCI o o o OH OH HI (55% in water) HI (55% in water) I OH OH Cl CI oO OH NHOH O N N H O NH O o ( NH2 Red Phosphorus Red Phosphorus HO Ho NH2 imidazole, MeCN imidazole, MeCN OImnidazole Imidazole / /(PhO)3, reflux NH NH (PhO)P, reflux

0

2023248067 10 H H O N N OMs N O OBocHN O O O BocHN BocHN,, O,, N N O BocHN HO HO N'N N Na 2CO 3,DMF, 80°C O O NN NOO NaCO, DMF, 80 °C

S (2) (2)

N NNN \\ N N (1) HCI /dioxane (1) HCI / dioxane

Cl CI - N- N O o OH OH TBTU, DIPEA, TBTU, DMF DIPEA,DMF

S S. NN N N O IZ N N H C T H 0-0 O N HO CI C1 ~N N TNN ( 0 NH NH 0 O

[0856] Step Step

[0856] 1: Preparation 1: Preparation of 2-amino-4-hydroxybenzoic of 2-amino-4-hydroxybenzoic acid acid A mixture

[0857] A mixture

[0857] of 2-amino-4-methoxybenzoic acid (1.0acid of 2-amino-4-methoxybenzoic 5.98 g, g, (1.0 5.98 mmol), mmol), red phosphorus red phosphorus

(556 mg, mg, 17.94 17.94mmol) mmol) 55%55% andand hydroiodic hydroiodic acid acid (10 (10 mL) mL) was heated was heated at 100°C at 100°C for 14for 14 ah in a h in

sealed sealed tube. tube. The The reaction reaction mixture mixture was pouredinto was poured into ice ice water. water. The The pH pHofofthe the solution solution was was adjusted adjusted to 6-7 to 6-7 by by sodium carbonate. The sodium carbonate. Thesolution solutionwas wasextracted extractedwith withethyl ethylacetate acetate (20 (20 mL mLx x3). 3). The The combined organicphases combined organic phaseswere were dried dried over over anhydrous anhydrous sodium sodium sulfate, sulfate, filteredandand filtered concentrated concentrated in in

vacuum vacuum totoafford affordcrude crude2-amino-4-hydroxybenzoic 2-amino-4-hydroxybenzoicacid acid (400(400 44% yield) mg, yield) mg, 44% which which wasinused in was used

the the next next step step without without further furtherpurification. purification.'HNMR (400MHz, ¹HNMR (400MHz, DMSO- DMSO- d): 7.53-7.55 d): 7.53-7.55 (m, 1H), (m, 1H),

6.12 6.12 (s, 1H),5.99 (s, 1H), 5.99- -6.02 6.02(m,(m, 1H). 1H).

[0858] Step Step

[0858] 2: Preparation 2: Preparation of 2-acetamido-4-acetoxybenzoic of 2-acetamido-4-acetoxybenzoic acid acid

[0859]

[0859] To To aa mixture mixture of 2-amino-4-hydroxybenzoic of 2-amino-4-hydroxybenzoic acidacid (400 (400 2.612.61 mg, mg, mmol) mmol) and imidazole and imidazole

(888 mg, mg, 10.06 10.06mmol) mmol)in in acetonitrile (20 acetonitrile (20 mL) mL)was wasadded added acetyl acetyl chloride chloride (789 (789 mg,mg, 10.06 10.06 mmol) mmol)

dropwise at 00 °C. dropwise at °C. The solution was The solution was stirred stirred at atrtrtfor 1010 for h and then h and quenched then quenchedby by water water (40 (40 mL). mL).

The mixturewas The mixture wasextracted extractedwith withethyl ethylacetate acetate (20 (20 mL mLx x3). 3). The Thecombined combined organic organic layers layers were were

washed withbrine, washed with brine, dried dried over over anhydrous anhydroussodium sodium sulfateandand sulfate filtered. Volatiles filtered. Volatiles were were evaporated evaporated in vacuum andthe vacuum and theresidue residue was waspurified purifiedby bycolumn column chromatography chromatography (ethyl (ethyl acetate/petroleum acetate/petroleum

ether = 2:1) ether = 2:1) to toafford afford2-acetamido-4-acetoxybenzoic acid(350 2-acetamido-4-acetoxybenzoic acid (350mg, 57% mg,57% yield).HNMR yield). ¹HNMR

306

(400MHz, DMSO- d): (s, 11.19 (s, 1H), 8.30 8.01-8.03 (s, 1H), 8.01-8.03 (m, 1H), (m, 1H), 6.92-6.95 (m, 1H), (m, 2.30 1H), 2.30 2023248067 10 2023

(400MHz, DMSO- 11.19 1H), 8.30 (s, 1H), 6.92-6.95

(s, (s, 3H), 2.15 (s, 3H), 2.15 (s, 3H). 3H).

Oct [0860] Step Step

[0860] 3: Preparation 3: Preparation of 3-(7-hydroxy-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine of 3-(7-hydroxy-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-

2,6-dione 2,6-dione

[0861] To a To

[0861] a mixture mixture of 2-acetamido-4-acetoxybenzoic of 2-acetamido-4-acetoxybenzoic acidmg, acid (400 (400 mg, 1.69 1.69 3- mmol), mmol), 3 aminopiperidine-2,6-dionehydrochloride aminopiperidine-2,6-dione hydrochloride(333 (333 mg,mg, 2.02 2.02 mmol), mmol), triphenyl triphenyl phosphite phosphite (2.0(2.0 mL) mL) in in acetonitrile acetonitrile(10 (10mL) mL) was addedimidazole was added imidazole(383 (383mg, mg,5.63 5.63mmol). mmol). TheThe reaction reaction solution solution waswas

heated to heated to reflux reflux for for10 10h.h.The Thesolution solutionwas was evaporated evaporated under reduced pressure under reduced pressure and andthe the residue residue was re-crystallized was re-crystallized (20% ethyl acetate (20% ethyl acetate in in hexane) hexane) to to afford afford 3-(7-Hydroxy-2-methyl-4 3-(7-Hydroxy-2-methyl-4-

oxoquinazolin-3(4H)-yl)piperidine-2,6-dione(110 oxoquinazolin-3(4H)-yl)piperidine-2,6-dione (110mg,mg, 19%19% yield).'HNMR yield). (400MHz, ¹HNMR (400MHz, DMSO- DMSO d 6): d): 810.94 10.94(s, (s, 1H), 1H), 10.51 10.51 (s, (s, 1H), 1H), 7.84-7.86(m, 7.84-7.86(m, 1H), 1H), 6.92-6.94 6.92-6.94 (m, (m,1H), 1H),6.85 6.85(s, (s, 1H), 1H), 5.16-5.20 5.16-5.20 (m, 1H), 2.73-2.85 (m, 1H), 2.73-2.85 (m, (m, 1H), 1H), 2.58-2.63 2.58-2.63 (m, (m, 5H), 5H),2.13-2.15 2.13-2.15 (m, (m,1H). 1H).

[0862] Step Step

[0862] 4: Preparation 4: Preparation of tert-butyl of tert-butyl (4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2 (4-(2-(2-(2-(2-(3-(2,6-dioxopiperidin-3-yl)-2-

methyl-4-oxo-3,4- dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)carbamate methyl-4-oxo-3,4-dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)carbamate

[0863] To a To

[0863] a mixture mixture of 3-(7-hydroxy-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6 of 3-(7-hydroxy-2-methyl-4-oxoquinazolin-3(4H)-yl)piperidine-2,6-

dione (161 mg, dione (161 mg,0.348 0.348mmol) mmol)andand 2-(2-(2-(2-(4-((tert 2-(2-(2-(2-(4-((tert-

butoxycarbonyl)amino)phenoxy)ethoxy)ethoxy)ethoxy)ethyl methanesulfonate butoxycarbonyl)amino)phenoxy)ethoxy)ethoxy)ethoxy)ethylmethanesulfonate (100 mg,(100 mg, 0.348 0.348

mmol,prepared mmol, preparedaccording accordingtotoprocedures proceduresof of similarintermediate similar intermediatedescribed describedininUSUS2015/0291562 2015/0291562) )

in in DMF (5.0mL) DMF (5.0 mL) was was added added sodium sodium carbonate carbonate (74 0.696 (74 mg, mg, 0.696 mmol).mmol). The mixture The mixture was stirred was stirred at at 80 °C 80 °C for for 6 h. h. The The resulting resulting mixture mixture was was cooled to rt. cooled to rt.Ethyl Ethylacetate acetate(30 (30mL) mL) was was added and the added and the organic layer layer was washedwith was washed withwater waterand andbrine. brine.The Theorganic organiclayer layerwas wasdried driedover overanhydrous anhydrous sodium sulfate, filtered sodium sulfate, filtered and and evaporated evaporated under under reduced pressure. The reduced pressure. residue was The residue was purification purification by by preparative TLC preparative TLC totoafford afford tert-butyl tert-butyl (4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo (4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methy1-4-oxo-

3,4- 3,4- dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)carbamate - dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)carbamate(55.4(55.4 mg, 24% mg, 24%

yield). 1HNMR yield). (400MHz, HNMR (400MHz, DMSO-d): DMSO-d): 10.98 (s,10.98 1H), (s, 9.081H), (s,9.08 1H),(s, 1H), 7.91-7.93 7.91-7.93 (m, 1H),(m, 1H), 7.32-7.34 7.32-7.34

(m, (m, 2H), 7.07-7.09 (m, 2H), 7.07-7.09 (m, 2H), 2H), 6.82-6.84 6.82-6.84 (m, (m, 2H), 2H), 5.20-5.24 5.20-5.24(m, (m,1H), 1H),4.24 4.24(s, (s, 2H), 2H), 3.99 3.99 (m, (m, 2H), 2H), 3.79 (m, 2H), 3.70-3.71 3.70-3.71 (m, (m, 2H), 2H), 3.56-3.60 3.56-3.60 (m, (m, 8H), 8H),2.79-2.87 2.79-2.87(m, (m,1H), 1H),2.57-2.70 2.57-2.70(m, (m,5H), 5H),2.17- 2.17 2.18 (m, 2.18 (m, 1H), 1H), 1.47 1.47 (s, (s, 9H). 9H). LC-MS: (ES*):m/zm/z LC-MS: (ES): 655.3 655.3 [M+H]*.

[M+H].

[0864] Step Step

[0864] 5: Preparation 5: Preparation of 2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2 of 2-(S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-

f|[1,2,4]triazolo[4,3-al[1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2 f][1,2,4]triazolo[4,3-a][1,4|diazepin-6-yl)-N-(4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-

307

Oct 2023 methyl-4-oxo-3,4-dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide methyl-4-oxo-3,4-dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)pbenyl)acetamide

(Exemplary Compound45) (Exemplary Compound 45)

[0865] To a To

[0865] a pre-mixed pre-mixed solution solution containing containing (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-

thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic hieno[3,2-f][1,2,4]triazolo[4,3-a][1,4|diazepin-6-yl)acetieacid acid(6.11 (6.11mg, mg,0.01525 0.01525 mmol) in mmol) in 2023248067 10

DMF (2.00 ml), DMF (2.00 ml), TBTU (7.34 mg, TBTU (7.34 mg, 0.02287 0.02287 mmol) and DIPEA mmol) and DIPEA(7.96 (7.96 µL, pL, 0.04575 0.04575 mmol) mmol) was was added 3-(7-(2-(2-(2-(2-(4-aminophenoxy)ethoxy)ethoxy)ethoxy)ethoxy)-2-methyl-4 added 3-(7-(2-(2-(2-(2-(4-aminophenoxy)ethoxy)ethoxy)ethoxy)ethoxy)-2-methyl-4-

oxoquinazolin-3(4H)-yl)piperidine-2,6-dione(8.46 oxoquinazolin-3(4H)-yl)piperidine-2,6-dione (8.46mg, mg, 0.01525 0.01525 mmol, mmol, prepared prepared by treating by treating the the productfrom product from step step 4 with 4 with HCl HCl in in dioxane) dioxane) and the and the was mixture mixture was left to left stir forto2 stir formixture h. The 2 h. The mixture was diluted was diluted with with ethyl ethyl acetate acetate and and water. water. The organic layer The organic layer was was washed washedwith withsodium sodium bicarbonate, bicarbonate,

water (3 x)x)and water (3 andbrine. brine. TheThe resulting resulting solution solution was filtered was filtered throughthrough a thin a thin pad pad of of silica silica gel gel and then and then

concentrated concentrated in in vacuo vacuo to give to give a crude a crude solid.solid. This material This material was purified was purified by silica by gelsilica gel

chromatography chromatography onon a Teledyne a Teledyne Combiflash Combiflash ISCO ISCO eluting eluting with with MeOH/DCM MeOH/DCM (0:100toto 7:93) to (0:100 to 7:93)

yield 2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3 yield 2-(S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-

a][1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4 a][1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-(3-(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-

dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide dihydroquinazolin-7-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide (10.1 (10.1 mg, 0.01077 mg, 0.01077

mmol,71.1 mmol, 71.1% % 1 HNMR yield).¹H yield). NMR MHz,MHz, (400(400 methanol-d4) methanol-d4) L 8.55 8.55 (s, 1H), (s, 1H), 7.96 8.00 (m,- 1H), 7.96 8.00 (m, 7.361H), 7.36 -- 7.50 (m,6H), 7.50 (m, 6H),7.03 7.03 - 7.09 - 7.09 (m,(m, 2H),2H), 6.87 6.87 (dd, (dd, J J = 3.03, = 3.03, 9.10 9.10 Hz, Hz, 2H), 2H), 5.22 (td,5.22 J = (td, 5.40,J10.91 = 5.40, Hz, 10.91 Hz,

1H), 4.704.74 1H), 4.70 - 4.74 (m, (m, 1H), 4.22J =(d,3.33 1H), 4.22 (d, J = Hz, 3.33 Hz, 2H), 2H), 4.10 (d, 4.10 (d, JHz, J = 4.30 = 4.30 Hz, -2H), 2H), 3.85 3.91 3.85 (m, - 3.91 (m, 2H), 3.79 2H), - 3.84(m, 3.79 3.84 (m,2H), 3.64- -3.71 2H),3.64 3.71(m, (m,7H), 7H),3.55 - 3.64 3.553.64 (m,(m, 2H), 2H), 3.42 3.42 - 3.50 3.50 (m, 2H), (m, 2H), 2.71 2.71 (s, (s, 3H), 2.66(d,(d,J J= =3.33 3H), 2.66 3.33Hz,Hz, 2H), 2H), 2.442.44 (d, J(d, J = 3.33 = 3.33 Hz,1.89 Hz, 3H), 3H),(s,1.89 3H),(s, 3H), 1.68 (d, 1.68 (d, JHz, J = 3.33 = 3.33 2H), Hz, 2H),

1.29 (br. 1.29 (br.s., S.,3H). LC/MS 3H). (ES*): m/z LC/MS (ES): m/z937.19/939.19 937.19/939.19[M+H].

[M+H]*.

[0866] Exemplar

[0866] Exemplar Synthesis Synthesis of Exemplary of Exemplary Compound Compound 44: 2-((S)-4-(4-chlorophenyl) 44: 2-(S)-4-(4-chlorophenyl)-

2,3,9-trimethyl-6H-thieno[3,2-f1[1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-((3 2,3,9-trimethyl-6H-thieno|3,2-f]]1,2,4]triazolol4,3-a]|1,4]diazepin-6-yl)-N-(4-(2-(2-(2-(2-(3-

(2,6-dioxopiperidin-3-yl)-2-methyl-4-oxo-3,4-dihydroquinazolin-8 (2,6-dioxopiperidin-3-yl)-2-methyl-4-ox0-3,4-dihydroquinazolin-8-

yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide

308

Oct 2023

O 0 COOH f NH HCI NH 2HCI o O COOH O OH (55% OHHI HI(55% in in water) water) OH AcCIN AcCl N IZ N O OH IZ H N NH2 NH RedRed Phosphorus Phosphorus NH imidazole, H NH 2 imidazole, MeCN O o o OH OH imidazole / (PhO) imidazole (PhO)P P 3

2023248067 10 reflux reflux

H O ZI H N o ZI o H o o o N OO N oO~~O~>< OMS ~NN OMs N BocHN O N N BocHN N N O 3, DMF, Na2 CO DMF, NaCO, 8080 °C°C O O

BocHN BocHN

[0867] This This

[0867] molecule molecule was synthesized using using was synthesized the same same method the method as described as described in Example in Example 1. 1. The keyintermediate The key intermediate was wasprepared preparedaccording thethe according scheme listed scheme above. listed ¹H 'H above. NMRNMR (400 MHz, (400 MHz, methanol-d4) methanol-d4) L 8.55 8.55 (s, 1H),7.96 (s, 1H), 7.96- -8.00 8.00(m,(m, 1H), 1H), 7.367.36 - 7.50 - 7.50 (m, 7.03 (m, 6H), 7.09 - 6H),- 7.03 7.09 (m, 2H),(m, 2H), 6.87 6.87

(dd, (dd, JJ= 3.03, 9.10 = 3.03, 9.10Hz, Hz,2H), 2H), 5.22 5.22 (td,(td, J = J5.40, = 5.40, 10.9110.91 Hz, 1H), 1H),- 4.70 Hz,4.70 - 4.74 4.74 (m, 4.221H), 1H),(m, (d, J4.22 = (d, J = 3.33 Hz,2H), 3.33 Hz, 2H),4.10 4.10 (d, (d, J =J= 4.30 4.30 Hz, Hz, 2H), 2H), 3.91 -(m, 3.85 -3.85 3.912H), (m,3.79 2H), - 3.84 - 3.84 3.79(m, (m, 2H), 2H), 3.64 (m, - 3.71 (m, - 3.713.64

7H), 3.55- -3.64 7H), 3.55 3.64(m,(m, 2H), 2H), 3.423.42 - 3.50 - 3.50 (m, 2.71 (m, 2H), 2H),(s, 2.71 (s,2.66 3H), 3H),(d,2.66 J = (d, 3.33J Hz, = 3.33 2H), Hz, 2.44 2H), (d, J 2.44 = (d, J= 3.33 Hz, 3H), 1.89 Hz, 3H), 1.89 (s, (s, 3H), 3H), 1.68 1.68 (d, (d,J= J =3.33 3.33Hz, Hz,2H), 2H),1.29 1.29(br. (br.s., s., 3H).3H). LCMS LCMS (ES*): m/z (ES): m/z

937.19/939.19 [M+H]*. 937.19/939.19 [M+H].

[0868] Exemplary

[0868] Exemplary Synthesis Synthesis of Exemplar of Exemplar Compound Compound 43: 2-((S)-4-(4-chlorophenyl) 43: 2-((S)-4-(4-chlorophenyl)-

2,3,9-trimethyl-6H-thieno[3,2-f|[1,2,4]triazolo[4,3-a[1,4diazepin-6-l)-N-(4-(2-(2-(2-(2-((1 2,3,9-trimethyl-6H-thieno|3,2-f]|1,2,4]triazolo|4,3-a|l1,4|diazepin-6-yl)-N-(4-(2-(2-2-(2-(1-

oxo-2-((S)-6-oxopiperidin-3-yl)isoindolin-4 oxo-2-(S)-6-oxopiperidin-3-yl)isoindolin-4-

yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)phenyl)acetamide

309

Oct 2023

o 0 o o o o O o NH NH CI CI ,O/- O NH NH MsCI, TEA MsCI, TEA O o NH NH NaNO NH NH o ~ NaN, NaN o -0 o- 0 o o O NaBH 4 DCM, 5oC DCM, 5C - DMF, 65 DMF, 65°C °C NaBH 0 OH THF, THF, -15 oC -15oC 0 OH 0 OMs OMs 0 o N3 o OH O OH o N 2023248067 10

Boc NH H NH Boc NH Pd/C, H , 40 Psi Pd/C, H, 240 Psi B NH HCI(g) 2N HCl(g) o CH 12 hh HN o HCI HN 30H,12 CHOH, dioxane, rt dioxane, rt HCI

Br NH o NO NH H2N HN¹¹ H O 02N 0 CHCN NH NH HH,2 ,Pd/C Pd/C N NH O HCI 07 + ON O 0 TEANreu 0C5 N- a o N 7=0 NN° o N° o HCI TEA, reflux

0 0 o o

Dess-Martin reagent o Dess-Martin reagent 0 N 0 02 N OH 02N o ON O CH 2CI 2 ON H CHCl H NH 2 NH NaBH3CN NaBHCN NH No NH 0 THF/MeOH/HOAc THF/MeOH/HOAc C N

o H 2N O\ H MeOH/THF MeOH / THF 0 HN 0-0 ZI

N N H2,Pd/C 0 H, Pd/C ' 0 0\ 02N \ / ON \H HN HN O-- O NH oOAO -N NO N o ZI Z, N N

O

[0869] The key

[0869] The intermediate key intermediate for preparation for the the preparation of this of this compound compound was synthesized was synthesized according according

the scheme listed above. The scheme listed The final final step step of of amide amide coupling wascarried coupling was carried out out under under the the same same condition as described condition as in Example described in Example 1.1. ¹H 'H NMR NMR(400(400 MHz,MHz, CDCl)CDC 3) d d 9.03 9.03 (s, (s, 7.45 1H), 1H),(dd, 7.45J(dd, = J=

8.71, 13.21 8.71, 13.21Hz, Hz,4H), 4H), 7.317.31 - 7.37 - 7.37 (m, 3H), (m, 3H), 7.24 7.24 (d, J =(d, J=Hz, 7.24 7.24 Hz, 1H), 6.84 (d, 6.84 (d, Hz, J = 9.00 J= 2H), 9.00 1H), Hz, 2H), 6.78 (d, JJ= 6.78 (d, 8.02Hz, = 8.02 1H), Hz,1H), 6.75 6.75 (br.(br. S., s., 1H), 1H), 4.73 -(m, 4.664.66 4.73 (m, 2H), 2H), 4.20 (d,4.20 (d, J= J = 2.74 Hz, 2.74 Hz, -1H), 1H), 4.07 - 4.07 4.12 (m, 4.12 (m, 2H), 3.80 -- 3.90 2H), 3.80 3.90 (m, 3.64-- 3.77 (m, 3H), 3.64 3.77 (m, 10H), 3.52 (m, 10H), 3.52 -- 3.58 3.58 (m, 1H), 3.35 (m, 1H), 3.35 -- 3.42 3.42 (m, (m, 3H), 3H),

2.68 (br. 2.68 (br. S., s., 3H), 2.52- -2.59 3H), 2.52 2.59(m,(m, 2H), 2H), 2.41 2.41 (s, 3H), (s, 3H), 2.02 2.02 - 2.08 2.08 (m, 2H),(m, 2H), 1.69 1.69 1.26 (s, 3H), (s, 3H), 1.26 (s, 3H). (s, 3H). LC-MS(ES): LC-MS (ES*):m/z 895.22/897.22 [M+H]. m/z895.22/897.22 [M+H]*.

[0870]

[0870] Protein Control Level Control Protein Level

310

Oct 2023

[0871] This This

[0871] also also description description provides provides methods for thefor methods the control control of protein with awith levelslevels of protein a cell. cell. This is based This is on the based on the use use of of compounds compounds as as described described herein, herein, which which are are known known to interact to interact withwith a a specific target protein specific target proteinsuch such that that degradation degradation of a of a target target protein protein inwill in vivo vivoresult will result in the in the control control of of the amount the amount of of protein protein in ainbiological a biological system, system, prerferably prerferably to a particular to a particular therapeutic therapeutic benefit. benefit. 2023248067 10

[0872] The following

[0872] The following examples examples areto used are used to in assist assist in describing describing the present the present invention, invention, but but should notbebeseen should not seen as as limiting limiting the the present present invention invention in any in any way. way.

[0873] Exemplary

[0873] Exemplary Embodiments the Present of theofPresent Embodiments Disclosure Disclosure

[0874] TheThe

[0874] present present disclosureencompasses disclosure thethe encompasses followingspecific following These embodiments.These specificembodiments. following embodiments following embodiments maymay include include all the all of of the features features recited recited in in a proceeding a proceeding embodiment, embodiment, as as specified. Whereapplicable, specified. Where applicable, the the following followingembodiments embodimentsmaymay alsoalso include include the the features features recited recited in in

any proceedingembodiment any proceeding embodiment inclusively inclusively or or in in thealternative. the alternative.

[0875]

[0875] An aspectofofthe An aspect the present presentdisclosure disclosure provides providesa acereblon cereblonE3 E3 ubiquitin ubiquitin ligase ligase binding binding

compound having compound having a chemical a chemical structure structure selectedfrom: selected from: X X X /GG X X X G X X G Q4 Q4 N 03 Q N N of Q Q;~ of /N Norry Z Z N N Z Z 02 Q2/ Q Q W W A A I- Q W W NN Q Rn Q Rn Rn Rn Rn R' R' G' G'

(al) (al) (b) (b)

G G

x X .1 N z Z X X X G X X X G Q N Q4N Q3 Q N Q4 Q3 N Z Rn 11~- Q2 1 N N Z Rn Q W NA Q W A N ZQ Q Y, Y Z Z R Rn Rn X X G' G' Rn Rn Q (c) (c) (dl) (d1)

311

Oct 2023 G G

X x N z Z X x x NJZAXx X 03 N X N Q4 Q4 N4 Q4 on on Q3 N Normal Z 2023248067 10 Rn

Q Q> N A R Q Q1 W Rn Z N A Rn Rn Q Rn Rn Rn

(e) (e) (f) (f)

G G

X N Z x .G.... X X X X G Q4 Q4 N/ Q Q3 oa Q N W Norm Z Z Rn Rn Q2 Q2 Q01 W W A A Q Rn Q Rn/ Rn Rn Rn Rn QR

(a2) (a2) (d2) (d2)

X X N/G G Q4 03 N "

03 Q Nurre Z

Q Q1 W W N A A Z

Q Rn Rn Rn Rn

(a3) (a3)

wherein: wherein:

W isis selected W selected from the group from the consisting of group consisting CH2 ,CHR, of CH, CHR, C=O, C=O, SO, S02, NH, NH, N, N, optionally optionally

substituted cyclopropyl substituted cyclopropyl group, group, optionally optionally substituted substituted cyclobutyl cyclobutyl group, group, and and N-alkyl; N-alkyl;

W isisselected W3 fromC CororN;N; selectedfrom each X is each X is absent or or independently selected from independently selected from the the group group consisting consisting of of O0 and andS;S; Y is Y is selected selected from the group from the group consisting consisting of of CH, CH 2-C=CR', , -C=CR', NH, NH, N-alkyl, N-alkyl, N-aryl, N-aryl, N-hetaryl, N-hetaryl, N- N cycloalkyl, cycloalkyl, N-heterocyclyl, 0, and N-heterocyclyl, O, and S; S; each Z is each Z is absent absent or or independently selected from independently selected the group from the group consisting consisting of of O0 and and S; S;

312

G and G' G'are areindependently independentlyselected selectedfrom 2023248067 10 Oct 2023

G and from thethe group group consisting consisting of of H, H, alkyl(linear, alkyl (linear, branched, branched, optionally substituted), optionally substituted), OH,R'OCOOR, OH, R'OCONRR", R'OCOOR, R'OCONRR", CH2-heterocyclyl CH-heterocyclyl optionally optionally

substituted withR',R',andand substituted with benzyl benzyl optionally optionally substituted substituted with R'; with R';

Qi, Q2,Q,Q3,and Q, Q2, and Q4 represent Q represent a carbon a carbon C substituted C substituted with with a group a group independently independently selected fromfrom selected R', R', N or N-oxide; N or N-oxide;

Aisis independently A independently selected selected from from the group the group H,(linear, H, alkyl alkyl (linear, branched, branched, optionallyoptionally substituted), substituted),

cycloalkyl, cycloalkyl, ClCl and and F; F;

R comprises R comprises -CONR'R", -CONR'R",-OR', -OR',-NR'R", -NR'R",-SR', -SR', -SOR', -SO 2 R',-SONR'R", -SO 2NR'R", -CR'R"-, -CR'R"-, -CR'NR'R"-, -CR'NR'R"-,

(-CR'O),'R", -aryl, (-CR'O)'R", -aryl, -hetaryl, -hetaryl, -alkyl -alkyl (linear, (linear, branched, branched, optionally optionally substituted), substituted), -cycloalkyl, -cycloalkyl,

-heterocyclyl, -P(O)(OR')R", -heterocyclyl, -P(O)R'R", -P(O)(OR')R", -P(O)R'R", -OP(O)(OR')R", -OP(O)(OR')R", -OP(O)R'R", -OP(O)R'R", -Cl, -F,-Cl, -Br,-F, -Br, -I, -I, -CF 3, -CN, -CF, -CN, -NR'SO2NR'R", -NR'SO2NR'R", -NR'CONR'R", -NR'CONR'R",-CONR'COR", -CONR'COR", -NR'C(=N-CN)NR'R", -NR'C(=N-CN)NR'R", - C(=N-CN)NR'R", -NR'C(=N-CN)R", -NR'C(=C-NO)NR'R", C(=N-CN)NR'R", -NR'C(=N-CN)R", -NR'C(=C-N0 2)NR'R", -SO 2NR'COR", -SO2NR'COR", -NO -NO, - 2, CO 2R', -C(C=N-OR')R", CO2R', -C(C=N-OR')R",-CR'=CR'R", -CR'=CR'R", -CCR', -CCR', -S(C=O)(C=N-R')R", -S(C=O)(C=N-R')R", -SF -SF and -OCF 3 ; and5 -OCF;

R' and R' and R" R"are areindependently independently selected selected from from the the group groupconsisting consisting of of aa bond, bond, H,H,alkyl, alkyl, cycloalkyl, aryl, heteroaryl, cycloalkyl, aryl, heteroaryl, heterocyclic, heterocyclic, -C(=O)R, -C(=)R, heterocyclyl, heterocyclyl, eacheach of which of which is is optionally substituted; optionally substituted;

n' integer n' from1-10; integer from 1-10;

' represents represents aa single single bond bond or or aa double double bond; bond;

representsa abond represents bond that that maymay be stereospecific be stereospecific ((R) or((R) (S))or or(S)) or non-stereospecific; non-stereospecific; and and Rn comprises Rn comprises1-41-4independent independent functional functional groups, groups, optionally optionally substituted substituted linear linear or branched or branched

alkyl (e.g., aa C1-C6 alkyl (e.g., linear oror branched C1-C6 linear branchedalkyl alkyloptionally optionallysubstituted substitutedwith with one one or more or more

halogen, cycloalkyl halogen, cycloalkyl (e.g., (e.g., aa C3-C6 C3-C6cycloalkyl), cycloalkyl),or or aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), optionally optionally

substituted aryl(e.g., substituted aryl (e.g.,ananoptionally optionally substituted substituted C5-C7C5-C7 aryl), optionally aryl), optionally substituted substituted alkyl- alkyl aryl (e.g., an aryl (e.g., an alkyl-aryl alkyl-arylcomprising comprising at least at least one one of an of an optionally optionally substituted substituted C1-C6 alkyl, C1-C6 alkyl,

an optionally substituted an optionally substituted C5-C7 C5-C7aryl, aryl,or or combinations combinations thereof), thereof), optionally optionally substituted substituted

alkoxyl group alkoxyl group(e.g., (e.g., aa methoxy, methoxy,ethoxy, ethoxy,butoxy, butoxy, propoxy, propoxy, pentoxy, pentoxy, or hexoxy; or hexoxy; wherein wherein

the alkoxyl alkoxyl may maybebesubstituted substitutedwith with oneone or or more more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl,

cycloalkyl (e.g., (e.g., aa C3-C6 cycloalkyl), or C3-C6 cycloalkyl), or aryl aryl (e.g., (e.g., C5-C7 aryl)), optionally C5-C7 aryl)), optionally substituted substituted

z Z of y x (e.g., optionally (e.g., substituted with optionally substituted with one oneor or more more

313

Oct 2023 halogen,alkyl, halogen, alkyl,haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., (e.g., a C3-C6a cycloalkyl), C3-C6 cycloalkyl), or aryl or aryl (e.g., (e.g.,

0 O X C5-C7 aryl)), optionally C5-C7 aryl)), optionallysubstituted substituted Yy (e.g., optionally (e.g., optionally 2023248067 10

substituted withoneone substituted with or more or more halogen, halogen, alkyl, alkyl, haloalky, haloalky, fluoroalkyl, fluoroalkyl, cycloalkyl cycloalkyl (e.g., a C3 (e.g., a C3-

C6 cycloalkyl),or or C6 cycloalkyl), aryl aryl (e.g.,C5-C7 (e.g., C5-C7 aryl)), aryl)), or atoms; or atoms; and and each ofx,x,y,y, and each of andZ zare areindependently independently 0, 1,0,2,1,3,2,4,3,5, 4,or5,6,or 6, n is n is an integerfrom an integer from1-10 1-10 (e.g.,1-4). (e.g., 1-4).

[0876] Another

[0876] Another aspectaspect of theofpresent the present disclosure disclosure provides provides a bifunctional a bifunctional compound compound having having the chemical the chemicalstructure: structure: CLM-L-PTM, CLM-L-PTM, or a pharmaceutically or a pharmaceuticallyacceptable acceptablesalt, salt, enantiomer, enantiomer,stereoisomer, stereoisomer,solvate, solvate,polymorph polymorph or or

prodrug thereof, prodrug thereof, wherein: wherein:

the PTM the PTM isisaa small small molecule moleculecomprising comprising a proteintargeting a protein targetingmoiety; moiety; the LL is the is aabond bond or or aachemical chemical linking linking moiety covalently coupling moiety covalently coupling the the CLM CLM andand thethe

PTM; and PTM; and the CLM the CLM isisa asmall small molecule moleculecereblon cereblonE3E3 ubiquitin ubiquitin ligasebinding ligase bindingmoiety moiety of of claim claim

1, 1, wherein when wherein when n isn 2, is 3, 2, or 3, or 4, then 4, then at least at least one one of R of or Rn ormodified W is W is modified to be covalently to be covalently joined to joined to

the the linker linker group group (L) (L) or or aaPTM. PTM.

[0877] In In

[0877] anyany aspectororembodiment aspect embodiment describedherein, described herein, the the CLM CLMisislinked linked to to the the PTM, the PTM, the

chemical linker group chemical linker group (L), (L), or or aa combination thereofvia combination thereof via W, W,X,X,R¹, R2,P R, R², R³, R4R', R, R, , R',Q1, Q2,Q,Q3,Q4,Q4, Qi, Q2, and Q5. and Q5.

[0878] In any

[0878] In any aspect aspect or embodiment or embodiment described described herein, herein, theisPTM the PTM is a moiety a moiety that binds that binds BRD4, BRD4, BRaf, EstrogenReceptor BRaf, Estrogen Receptor(ER), (ER),ororAndrogen Androgen Receptor Receptor (AR). (AR).

In any

[0879] In any

[0879] aspect aspect or embodiment or embodiment described described herein, herein, the compound the compound may further further comprise may comprise a a second E3ubiquitin second E3 ubiquitin ligase ligase binding binding moiety moietycoupled coupledthrough througha alinker linkergroup. group.

[0880] In any

[0880] In any aspect aspect or embodiment or embodiment described described herein, herein, the second the second E3 ubiquitin E3 ubiquitin ligaseligase binding binding

moiety binds moiety binds or or targets targets an an E3 ubiquitin ligase E3 ubiquitin ligase selected selected from from the the group consisting of Von group consisting Hippel Von Hippel-

Lindau (VLM),cereblon Lindau (VLM), cereblon (CLM), (CLM),mouse mouse double-minute double-minute homolog2 homolog2 (MLM), (MLM), and inhibitors and inhibitors of of

apoptosis apoptosis proteins proteins (ILM). (ILM).

314

[0881]

[1880] In any In aspect or any aspect or embodiment described embodiment described herein, CLM theCLM herein,the is represented by by is represented a chemical a chemical

structure selectedfrom structure selected from thethe group group consisting consisting of: of: o 0 o 0 00 o o o N HNH HN =0 NtNH =0NtN N o N O

2023248067 Rn Rn o Rn Rn o o 0 o 0 O 0 o NHN HN NnNH HN

NNO 0 N o N 0 o

Rn Rh Rn Rn o o o O o F EL

EF 0 0 0 0 0 o o o o o NH N Nt H HN HN N N N 0 o / N 0 o N N

Rn Rn Rn Rn 0 o 0 O o o 0 o N HN NHH HN N o N 00 N O Rnt 0 Rn o o Rn Rn '

0 0 1,0 0 o O o o NH HN t NH HN

Nt N N =0o N N =0 O

-- 'Rn Rn 0 o

315 E15

Oct 2023

o o 0 o 0 o 0 o

NH NH NH 2023248067 10 NH 0 NN 0 NN o o

Rn Rn O Rn Rn O o o ZI H o o N N 0 o 0 o 0 o 0 o NHN NH N N N 0 o

Rn Rn 0 Rn Rn oo o 0 o 0 o 0 o N NH NH N NH NH N N N N =0 N o N o NN N Rn Rn 0 Rn Rn O o o ZI

0 H N 0 o N O 0 x X o

NH NH N N0 N 0 o

N Rn Rn Rn Rn

[0882] any aspect In aspect

[0882] In any or embodiment or embodiment described herein, herein, described the linker linker the (L) (L) comprises comprises a chemical a chemical

structural unit represented structural unit representedby by thethe formula: formula:

-(AL)q_ -(A¹)q-

wherein: wherein:

(AL)q (A¹) isis aa group groupwhich whichis isconnected connectedto to at at leastone least oneofofthe theCLM, CLM,thethe PTM, PTM, or a or a combination combination

thereof; thereof;

qq is is an integergreater an integer greaterthan thanor or equal equal to 1; to 1;

316

Oct 2023 each ALisis independently each AL independentlyselected selectedfrom fromthe thegroup groupconsisting consistingof,of,a abond, bond,CR¹¹R¹, CRL1RLO, 2S, O,SO, S, SO, SO 2 , NR¹, SO, NRL', SONR¹³, SONRL 3 , CONR¹³, SO 2 NR, SONR¹³, NRL3 CONRL 4, NRL³SONR¹, CONRL', NRL³CONR¹, NR SO 2NRL 4CO, , CO, CR¹¹=CR¹²,2 ,C=C, CRLl=CRL SiRL'RL 2,P(O)R¹¹, C-C,SiR¹¹R¹², P(O)RL, P(O)OR¹¹, NRL3 C(=NCN)NRL P(O)ORL,NR¹³C(=NCN)NR¹, , NRL 3C(=NCN), NR¹³C(=NCN), 4

NRLC(=CNO2)NRL NRL³C(=CNO)NR¹, 4 , C3-llcyclalkyl C3-11cycloalkyl optionally optionally substitutedwith substituted with0-6 0-6R¹¹ R and/or RL 2 and/orR¹² 2023248067 10

groups, C3-Iiheteocyclyloptionally groups, C3-1heteocyclyl optionallysubstituted substituted with with 0-6 0-6 R and/or R¹¹ and/or RL 2 groups,aryl R¹² groups, aryl

optionally substituted with optionally substituted 0-6 R¹¹ with 0-6 RL and/or RL2groups, and/orR¹² groups,heteroaryl heteroaryloptionally optionallysubstituted substituted 2 2 independently are optionally with 0-6 with 0-6 R¹¹ RL and/or and/orR¹²RLgroups, groups, where where RLR¹², R¹¹ or or RL each, each independently are optionally linked to other linked to othergroups groups to form to form cycloalkyl cycloalkyl and/or and/or heterocyclyl heterocyclyl moiety, optionally moiety, optionally

substituted substituted with with 0-4 0-4 RL groups; and RL groups; and 2 RL³, RL, RL RL¹, R¹²,, RL 3 , RL4 andRL R¹ and RLare, are,each eachindependently, independently,H,H,halo, halo,C-alkyl, OC1-alkyl, C1-8alkyl, SC1-alkyl, OC1-8alkyl, SC1-8alkyl, NHCi-8alkyl, NHC-alkyl, N(Ci-8alkyl)2, N(C-alkyl), C3-lcycloalkyl, C3-11cycloalkyl, aryl,aryl, heteroaryl, heteroaryl, C3-11heterocyclyl, C-heterocyclyl, OC1- OCi 8cycloalkyl, SCi-8cycloalkyl, scycloalkyl, SC-cycloalkyl, NHCi-8cycloalkyl, NHC-cycloalkyl, N(C1-8cycloalkyl)2, N(C-cycloalkyl), N(Ci N(C-

8cycloalkyl)(C1-8alkyl), OH, OH, scycloalkyl)(C1-salkyl), NH 2NH, , SH,SH, SO2C-8alkyl, SOC-alkyl, P(O)(OC1-alkyl)(C1-alkyl), P(O)(OC-alkyl)(C.alky1),

P(O)(OC1-8alkyl)2, CC-Ci-alkyl, P(O)(OC-alkyl), CC-C-alkyl, CCH, CCH, CH=CH(Ci-8alkyl), CH=CH(C-alkyl), C(C1-8alkyl)=CH(C1-alkyl), C(C-alkyl)=CH(C-alkyl),

C(Ci-8alkyl)=C(C1-8alkyl)2, Si(OH) 3Si(C-alkyl), C(C-alky1)=C(C-8alkyl), Si(OH), , Si(Ci-8alkyl)3,Si(OH)(C-alkyl), Si(OH)(C1-8alkyl)2, COC1-8alkyl, COC-alkyl,

CO 2 H,halogen, COH, halogen, CN, CN,CF 3 , CHF CF, 2 , CH CHF, 2F, NO, CHF, SF,SONHC-alkyl, N02, SF, SO2NHC-salkyl, SO2N(C1-8alkyl)2, SON(C-alkyl), SONHCi-8alkyl, SON(C-alkyl), SONHC-alkyl, SON(Cl-salkyl)2, CONHC-alkyl, CONHC1-8alkyl,CON(C-alkyl), CON(C1-8alkyl)2,N(C- N(Ci 8alkyl)CONH(C1-8alkyl), N(C1-8alkyl)CON(C1-8alkyl)2, alkyl)CONH(C1.salkyl), NHCONH(Ci-8alkyl), N(C-alkyl)CON(C-alkyl), NHCONH(C.salkyl),

NHCON(Ci-8alkyl)2, NHCONH, NHCON(C-salkyl), NHCONHN(C-alkyl)SONH(C-&alkyl), 2, N(C1-8alkyl)SO2NH(C1-8alkyl), SO2N(C1 N(C-8alkyl)SON(C- N(C1-alkyl)

alkyl), NH SO2NH(Ci-8alkyl), 8alkyl)2, NH SONH(C-salkyl),NH NH SO2N(Cl-8alkyl)2, SON(C-alkyl), NH SO 2NH 2 NH SONH. .

[0883]

[0883] In In anyany aspect or or aspect embodiment described embodiment herein, described herein,the theALALisisselected selected from from the the group group consisting of: consisting of:

-N(R)-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-, -N(R)-(CH2)-O(CH2)-O(CH2)-O(CH2)p-O(CH2)q-O(CH2)-OCH2-,

-0-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-, -O-(CH2)-O(CH2)-O(CH2)-O(CH2)p-O(CH2)q-O(CH2)-OCH2-, -0-(CH2)m-O(CH2)n-O(CH2) 0-0(CH2)p-O(CH2)q-O(CH2)r-O-; -O-(CH2)-O(CH2)-O(CH2)o-O(CH2)-O(CH2)-O(CH2)+O-; -N(R)-(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-O-; -N(R)-(CH2)-O(CH2)-O(CH2)-O(CH2)-O(CH2)-O(CH2)-O-; -(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-O-; -(CH2)-O(CH2)O(CH2)-O(CH2)p-O(CH2)q-O(CH2)-O-; -(CH2)m-O(CH2)n-O(CH2)o-O(CH2)p-O(CH2)q-O(CH2)r-OCH2-; -(CH2)O(CH2)-O(CH2)-O(CH2)-O(CH2)-O(CH2)+-OCH2-;

-(CH 2 )mO (CH 2)r- N N - (CH 2)oO (CH 2)p ;

317

-+- jICH2 )m-N \/N-(CH 2 )n-NH :

-+- -1CH 2 )m-N \/N-(CH 2 )n-0 :

-:-(CH 2 )mO(CH 2 )n-N N-CH 2 )o-NH :

-|- 4(CH 2 )mO(CH 2 )n-N N-(CH 2 )o-O :

-|- --4(CH 2 )mO(CH 2 )nN&\N(CH 2 )o-NH :

--±(CH 2 )m(CH2)nNN\NNICHA)oO -|-

-|-"(²HO) (CH 2 )m-: N-|- -:-N 0 O N N N-|- -'-NO 0 O : N\ N :

-!-"(ZHO) ("CH 2)m4

(CH) -CH 2)m/N (HO) N (H) -: 4: :

2 )m N-,, N :

T CN-CH N N N\ N' (CH, : O : (HO)

o O

0_ 0 N N N N

O o

N/ N- N NF \N-N

0 o" N N

N N N N

O o

^^^^

N -CH 2)mO (CH 2)nO(CH 2)pO(CH 2).0 N N\/N :

318

HN HN / O(CH 2 )mO(CH 2 )nO(CH 2)pO(CH 2)qOCH 2 O(CH)mO(CH)O(CH)pO(CH)qOCH x X -:-NH NH OC

\/ O(CH 2 )mO(CH 2)nO(CH 2)pO(CH 2)qOCH 2 O(CH)O(CH)O(CH)pO(CH)qOCH

NH NH O(CH 2 )mO(CH 2)nO(CH 2)pO(CH 2)qOCH 2 2023248067

'r\ O(CH)O(CH)O(CH)pO(CH)qOCH

_ NH -:-NH \/ O(CH 2)mO(CH 2 )nO(CH 2 )pO(CH 2 )qOCH 2 O(CH)O(CH)O(CH)pO(CH)qOCH ;

NH -:-NH \/ O(CH)O(CH)OCH O(CH 2 )mO(CH 2)nOCH 2 ;

O(CH 2 )mO(CH 2 )nOCH 2 O(CH)O(CH)OCH HfN HN X X ;;and and

''NIZ N \ H / N-(CH 2 )mOCH 2 H N-(CH)OCH N N ;wherein ; wherein

m, n, o, p, q, and r of the linker are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, m, n, o, p, q, and r of the linker are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,

15, 16, 17, 15, 16, 18, 19, 17, 18, 20; 19,20; when the number when the numberis iszero, zero, there there is is no no N-0 or0-0 N-O or 0-0bond bond R of the linker is H, methyl and ethyl; R of the linker is H, methyl and ethyl;

X of the linker is H and F X of the linker is H and F

'N IZ N \ H / N-(CH 2 )mOCH 2 H N (CH)OCH N N where where m m of of thethe linker linker can can be3,2,4,3,5;4, 5; be 2,

319

IZ IZ IZ H H IZ IZ H H ZI IZ N H H

0 N IZ IZ

H HN NN I H-, ZI N

o IZ HN O NON 0-: -- i o o ZI <<<<<<<<< IIIIIIIII.

O N N N N o

N N

32

Oct 2023 IZ IZ H H H ZI H

N ZI 0 ''N ZI '. Q HH H H 2023248067 10

IZ ~N~ IZ IZ H H H H

ZI /N No IZ H H H H 0 O= o 0,0 O S S S s IZ ZI ZI o 0 H H H H H H

ZI N H IZ /N /NZI H H H H

IZ ZI N N O O N O H H HH O ZI O HN HN N H ' N T HN N -N,0

IZ N N N HH AH ZI N N "N-~ ZI H N H H

IZ N N N ZI H H H 'N ZI H H O H N ZI N NH N N H IZ N H

ZI 0 H_ H ZI N N NH H O

O HN N o N H21 O HN N N n

3:5 2023248067 10 Oct 2023

ZI HX H

3 oo o ~o

OH Ho

O O

o/ O

O IZ IZ N N O H

Kz~o~322

error

mai, maral, 0 0 somes N N

0 2023248067

ress N N ; swag 0 nN 0 ; Onaly 00 N

Osraly N Ronal ; Onaly 0

0 0

Onaly Oraly ; N NN N NN0

0N-A o - -,,

analy. Onaly. aor N'p N N

N-N 0 0 -o N N ;

N N N N N NN N N N ; yyy

o 32 N N N N N N in

N ; HO HO

0O N 0 O N N in N N s ? ; O ZNN ,,,

O 3 O 333

O 3'm /\NN N-- N N-0 N -I-Nr in N N-- N viv N -- N N N N ; ; ;

viv / in N N N NIn N N N

324

HN ZI ZI H N N H H H H x X

HN N'-"""/O~NO ZI ZI N N N H H H /7 X=H, F X X X=H,F H IZ H IZ H H NN NN~- N N ,4"C ' N ZI N N H N

IZ H ZI

N-s~ N0 I\ NH ZI H N N 0 H ~ NH IZ

HN H N ZI O

ZI ZI ZI N H N o x x

IZ HN ZI

H N ~.11"I X X H~~ N S

ZI N ZI N xx ZI H H H N N X H '~' I N IZ N H ZI HN HI N N 0 H N N N

NN N~x X X H - 0_ '\

NH NH NN N N

325

-:-NH NH -'--NH NH N--H N N N NH o N N N O -:-NH0 -i-H -N-NH \/ N 0 N-\ O NH O NH NH N N o N N -'-NH __ _ ,- -:-NH --- i-H N 0H~ O ___ I__ 0_ __

NH NH NH o O -:N _ -\ -:- H -I N N 0'-1 NH o ZI H ZI N N ~~H~ O ~ NN\ H

o ZI H N ZI H N IZ IZ N 11111

S c \\O H N 0 'NrN H o H N N

fl N X HN HN HN /0 N X X X=H,F X=H,F x X

HN o HN HN N N N N '-N N

HN 0\ HNm-<)0 HN HN N O N

X IZ millo X IZ

N N

N NN N o N N N N N N N N N N\-N

N N o O NN

326

O O HN 0- H N"K1"0- HN N4 O ~a HN N N N N

HN /\HN-O k \N-O" HN~K HN ~'0N Y\\ HN N xX 2023248067

IZ HN N H ~ IZ HZ N H H x X x X

O IZ HN IZ

-H N H \, HH x0 X

H ZI H NN ZI H ZI N N H N H"

HN-0

H N 00 -o HN- - NNK HN HN N O HN N

7HN H ~ ~ N N '~ HN HN N'.,'\ N HN N O N N N N IZ N IZ N H H N H o

-'-N N N N- N:N N-C -|-N - ' -- N N- -N N N N N

327

-|-N N N N N N N N N :NX - N- :N~XN- N-' 1111.

O NJ - HO Ho HO Ho N N -:-N -|-N N -- N N N -NN 0- N N N

' -|-N -:-N \/N- O -- N -|-N N- O N O N N

0-~~0 O N -- N N N _- -|-N NO -- N 0ON N HN O

-|-NN O NN H --N HN N N O N N'-- \ O

/ o O -N -|-N N N N HN N N OHN N HN HN N O

HN N HN N o N N N N HN N N N

NH O HN N O o HN N O N

1 HN O N,1 1 , N ZI H -NN o N HN 7-FN F F F where each where each nand n and m of mof the linker the linker can independentlybe can independently 0, 5, be 0, 1, 2, 3, 4, 1,2, 6, 3, 7, 4,5,6,7, 8, 9,10, 8, 9, 10, 11, 12, 11, 12, 13, 14, 15, 13, 14, 15,16, 16,17, 17,18, 18,19, 19,20. 20.

[0884] In In

[0884] anyany aspect aspect or or embodiment embodiment described described herein,the herein, theALALisisselected selected from from the the group group consisting of: consisting of:

328

N N \-iN-V ,.../N N N N

' N N N N N oN N ~ N N - N

N N O-/- NN'Y C N-' N N N N N-NNN,'N\ N N

,"'N N. ' 01 -' N N N N N N N N

* 0-/N N", N N' N,, N N N N N u N N N NIN w

N N N N NN N N N N N

N N N N N 4NlaoN N N N Hm

HN smashs >0 N" /ZN'.">N 0 ~/329

o0 0 Na H H

F00 F

NN H garanal, H

2023248067

laranage laranage N

o N:7a

0

0

N N 0

N NH H HN

NH granaly N0 HH

NH granal NH smorge H 0

H H

manal mail, storal NH -. 0 0

H H

0 NH scharal s

H IH H o o ZI IZ N - 00

ZI N N

330

sociors songs langs goods H IH 0H

F0 -,.0,O z~ N( N -,_,O N

0 leases soars smorges H H

,yO N N .,0 N 0 N

givens has lagues H 0

H ' F

your governy yours gibbles grows yourds /H 0- ' 0

N.I H 0 N-

H-- N 0 00

H

0

smally sons glorms 0 00

H /NN NN KK/-~~NN F H Hillons gooses solls 0 / N.~ 0 0,0N' N. \~~ N.~ HIH~ H

shows in in sogars H

0; 0 NF 00

ND 0 H'- O / H H H 0 F H

nass shows grants 0 /.o N 0 0

"'N'o 01 0

0 0 H H.

N raos grobas N ~ 0 H E N0 0 F H H H

gobbles ~0 goods Harry iasos N0 00 0

HH 0 0

grobies subses story H~ 1 N'' /ql N1 oNF 0 F N F

I~/ N .

0l o0 N'N' 0N.0 0 H

Has HOUSE HF

331

Oct 2023 0

0~~~ 0-o oq o''' ZI H N N HN N H H I 0 F H

IZ N 2023248067 10 H ZI ZI ZI F F F F H

O o 0 Ns .r-Y ZI N IZ N N ZIN H H ;H ad and Hwherein H , eachnm wherein each m and n nis and is

independently selected independently selected from fro0, 1, 2, 0, 1, 2, 3, 5, 6, 7,6,7, 4, 3,4,5, 8, 9,8, 9,10,11, 10, 11, 12, 13, 13,14,15, 12,14, 16,17, 15, 16, 17, 18,19, 18, 19, 20. 20. 108851 In any

[0885] In any aspect aspect oror embodiment embodiment described described herein, theALA'is herein,the is selected from the selected from the group group

consisting of:. consisting of:

O O O ZI N O N N H H N'' H O ZI N O N N O H H O H * N N H ZI N H H

H ZI H-N ZI N O N O H H Lr~ 9N U -'-- IN O IZ N N H N H

NNI %0 o NN K NN 0 O N N 0." 0./

o

o NO0ON N N 0 N N N N O o

332

N N N N N N IZ H O N NI N HI N N N ZI IZ H H N N N N ZI H 2023248067 N0_ 1-- N '0%%

N N iii~~ N ZI HNN~ N N H N N N. N-XO N ,*F F N ZI ZI N H H

N N H H N N F F FF N NfN F F F F H F FH N N N N N N

ZI ZI N N H H FF FF N ZI FEH FE H;F O N N H FF ZI N N N' H N FF FF N FE F* FF N N N ZI N H FF FF

333

N N N N N ZI ZI FEF H H FF N

ZI ZI o H. N H. H

ZI H H H IZ

OH OH Ho Ho 0 O ZI N OHH Ho IZ HH H H OH .~ Ho OH Ho ZI

0o H O

O H. ZI

o 0 H IZ H HH IZ OH H O O 0 O ' O ZI -o -N" H 0 0 ZI O"H ZI N

ZI 00

H ZI ZI '

0'. HH HN

334

m010 n NN XXXXX n n

0

tax n n n o m Y %' nnn 00 N

%%N N -- - n o 0 0 0 N N

o n o XJH o ZI ZI

ZI

N N NN pmn n

ZI

N N n nn p n

NS~ / N -\ N NN nw %n

n n p N N XN: m m N N N N

/\ n N n-"h' N /

m m

NN N N N 0 m m "

Na 0 m N N o

N Na o m N m N -\/ NH7--"

O NH N N -'N n ~ N\ N N\ N

I + N m N N m N N m

335 o O -NH N N--CN N N N m m m 2023248067 10 Oct

N - ~ N m- -N m

o N

N N N N I n $ n

o Im m

0 0 N ZI N NH m H n

N N N N m n n mN N

N m n N N. m

N N N -0 n m n N N

N N N N m

0/ N N N N N n n m m

N N m

0~* " 33N

N N N N m n m N

Nm - N ?~ >NN m1- - N m who

N + N N + N m N m m -/--- m N

N + N + N

m N½ I N N r r~N N

N N

m WE N 73/25

N N N. N N

m m mm m WE N WE N N N N N N N

m m

N3N

3/2 0½ N

N N 0 m m .m m f m m OH N N 0

N N

N N m f

CF N OH N N N

N N I -o

N N

NoN

N n NC N --+--- N to 0n N

m N, 0 ()N\0 N WE N-"'fl 0 NC N N N n 'M

m N N OH

N 0 ,

N n N'N-: m m N N 'N N

0N N mN

N + m "ON m Kn)

N N N m N

CL. ZI mk, 3/2/212 mm N

N N N m m m

I- 338

Oct 2023

N N N n NN N N N m IN/ N m 2023248067 10 N

N IH n

m NH N n NH NH2 N N

o m N N n o m

N NN N o o m n 0, 0 ---0

n N N

o o N N m HO N N p o N N n HO n m m o Po~W>) p n m HO N N N, mN N n n m 00

N N

N N N 33HO m n

H

0 Z N Z y HN

IZ NN N N N N N N HH E NN E E E

0 m H -\N w

N/ N ON/ N E ~W

N N u N N

N\N w N

N N N N

340

Oct 2023

N )emN 0-k0 m N \N- N N0 N N

-N N N- 2023248067 10

m N N + N -N m N- /n ~r"" -:-o0

X N N"\ N N - N-:

X N N m n N//L~ m m N N n X m N N :\ -H 0 o O ti

X N N N IZ m

340

N N N N''~ NNI N u n C:Nu N N N N

N N NN N N N N,, N N w N. I N N ~-,I N -, N\-~'N~.-N N - 0/ u N N N N N N w N N N

N N' N N N N N-' N IN NN N N

u N N NNN N NN N N mm w w

- ~~\ b/-\N Na -N"' N N N N N N N N

N N N N N N N % % -:N N NNNI N w N

N-N N N N N N N N El- u N N u

0-÷ -:-N N N N u N N u

CFE

N N N N w N N N

342

O O O O o 00 0 o 0-11 m n p

o m O n O 0 o

n o m H ZI 0 H QQ0 o o o /'1 0 o N 11

m 4n / m% n m m ZI H o O o o N

0~~ 0 0 N N m m n ZI

o o O O N HH n o p m ZI H o I'-f O O N

mm n m n n oo p

000 o N 0 0 n O no m mn m n 0o m O O o o IH

n o p q m o O o o

nn n o p m ZI

Oo oo o0 ,, o0%

mn n 0 q m m HN

o O o o

o0 m n m 0 n ZI 'I o O o O

n o p q m m n 343 ZI

O o o

n o p m

o N O o

m n 0 p

N

11 N rlN N n N

"HX

-1°F2 N".

0 ~

N

****** the N <0-0+ N + m N

+Q-0+ + N Our +040+ N0,

0'l

N _

0_

M

N \ 0

things NN

0,1

N

344N

N N +0-0+nx NN N / NN N N

X N N N

o~ N' N

N N N

0 N

_ - N - :- _ NN

ro o

0 m

34 X

H) H2

IZ m N m N N N m n

o O N N N N N-- o 6H) NH-- NH o n n m m 0 m m4n N N m N N n m ......!! IIIIIIIII.

N N N N mNO Ni -:-o\

O NN N N N N n m o

N /--N N N-I-- - : N N N N o 00 o

-/ X/\ N N mNKXV/ 0 n N N -- :-o N N o O n N

N mNI346 N

N N N n n m

N N m n N N n o

- 0 I N o N N n 2023248067 N N

0 o N N 0_/170 n N N m m n m o o N X NH

&MN N lm \,N l -\ 0

IIIIIII

0-1 NN NH q N N nN ..........

HN m N m O o n N m

X NH X NH

o m N N N-NN n 0 O N H n X O m N n

m X 0)m N N N n N N N N

347--

0N~NN '0 'A'

N o o m m 0 -' " -N -N N N

m m 0

n 0

N 0

0 0 0 m m m N 0 n ' 0 N 0 ' -' p.. 0 N NN 0' n m -'N 0 0 N N

N,' ifi

0x m ' 0

you --/-- \A 0~~0O

N N 0

0

N,

N ~ ['N0

N ' N.0 N,' -,

A, N 0 0

n

N, ~ 0 N 0 0 N,' ("'N

0 ~ 0 A"~NN,,A' 0 0 0 N

~',~0 0 /

'N N 0 o o

I /'~'-0

---/--

>0 0 II

n 0 m

m N

N H 0

the Angust , 0 0 0

N'

348

N

0 N

n nn

0'

0",O

0 N0 to they n0

0 n 0 N

name 0 N

' 0 0-N

0- I Ir

This NN 0 NN 0 0=

think ,,- 00o-4 0 NN

00

00

0 0

349

NC

N 0/ N to

And 00 00 -\Y

N

2023248067

o-t Not

00

00N

0' 0

N o-t ON N 00 NC

0 NN It n 00

0 ---. 0CN' I N n

/ 0 0

-+- N 3 0 N o

N N

0

NC NC '00

350

0' ',0 0 nF F

2023248067 10 Oct

the F FF

F. F

00

thomx "0

N CF N CF

N..,, N/ N F3

F3C N 00

nong N

F 0 0

m N

any N

351C

socialy

CF 3 Oct 2023 CF

m m m m n 0n n

OH

2023248067 10

*0o

n

OH

+ m mN n

0

N

0

N

352-

00

N

00 00 N. l-5'

2023248067

NC

N 00 N

N ~ , oxl

0 NN

N

'0 N

F3C

NNN

F 3 CN

0 0 ,0N N,>5 N F3C

N

00

N N o N NO N0N N N.

NC NN

05 HO NN

N

N 0 N N

N ,,>~ 0 --- O N CN

0 C

N N N N N N

353

00

04 .N. N 0> '00

CF 3

00

0 N 0

CF

0'

141 " N

'00

CF

"'O0

0

-- N

00

oxo '00

'0O.

N.

Nc

NC

o' N N 0>

354

N N 0N N°

N NN m / 0 F3C NJ

N N N 0 N NN

N N0

N

+0+1.0+ I~~~0

N N- N N. m 00 n

-11 1N N m0N

* 00 !-!----------------------

NJN N

- 0 N N 00 N

N O

0 o0 N m 0 N 0N

0 00 N.

0 N N

HO

.0 N 0

N 0 \10N 0 HOJ

N N

3555

N N NN r, r 4-,m Nj N ,N N 0O 0 %%

m nn p p

N N N rl* N r r j 0o N N %N N 0 q N m n n pp

N N N N N N

m n

N \\, N0 N

mm N / - NH2

N N N o o n m n

X + N

O m n

N N

N N N5N

m n p

H N ~ N N' HN

0rH N NN III

0m N o

H IZ ,, N N N N'" HN H NN

N 0 o N

H IZ S

N N N0 0'

N NN

N IZ 0 0 0" N N u N o N N

N N N N NN N NA IZ N N N N

N j

N N N N

HN 0 N N rN

N N N N N N N N N O% N.,'O* ,., CN, CFE //N

357 ESE

2023248067 10 2023

0 IN" O% N Oct O ,wherein , wherein each eachm, m, n, o, p,q, n, o, p, and r is rindependently q, and 0, 1, 2, is independently 0, 1, 2, 3, 4, 5, 5, 6, 6, 7, 7, 8, 9, 10, 8, 9, 10, 11, 11, 12, 13, 14, 12, 13, 14, 15, 15, 16, 16, 17, 17,18, 18,19,19,oror20.20.

[0886]

[0886] In any In any aspect aspect ororembodiment embodimentdescribed herein, described the AL herein, theisAL selected from the is selected fromgroup the group consisting of: consisting of:

- O- - O~~- O f a--.' \-/'' O --- 0\-- O 0 0 O - O - O 000 - O O O

O O 358 o /--NH NH 0 HN- HN O o HN HN---. 0 O 0 O WI'\Q O O ;

0 0/0 0 0 0 NH' NH 0 0 0 HN---* HN O

NH o O ; ;

00 0H ; ; ;

O ; ; ;

00 Ó

0 0--*-\ 0 0

0 0 - - 0 0 0 o -- ' -\/000-

0 0 0 0---\/ o 0 0-'

358

Oct 2023

2023248067 10 0 0 0 0~0 O

_ -- _ 0

N 0 00 0 \d

O. - N 0 0-N N N

0 0 --- 0 -- 0 0 -- \ 0 0 0 N N N

N N N N /

N 0--.

N N N N 0 0- 0 0 0

N N -- N -- N N30-

N N N N N N N

N -N N\NNN 0- N

~ 0~NN --

2023248067

-C -/NN N --- N CN_-ND- N N N

N NN N -- CN N -- CN-P N -- CN-CN---.,-_f\\- N N NH NH 0 0

0- HN O O o 0 NH O O 0 0 0 0 NH O O O 0; 0 O 0 0HN O HN

0 000 O o NH NH O O o O 0 0

HN 0 O NH 0 0 0 O 0 _N

o 0 --- 0 0-\-N HN--- N -- '0 0 ~ -- O 0~' 0 O HN 00

NH HN O O 30

O 0 HN--- 0 2023248067 10 Oct 2023

- - HN O - NH-\0 NH NH O NH O O O ;

00 O O NH 0 -N\ / C HN HN --- -/- O OHN-- O ; O HN

O 0 O 0 O -- \ / 0 O 0 O HN--- HN -- 0 O HN ;

- o0 0 O NH O O NH O ; O ;

O NH NH0\ I0 NH H

0 N O ; O ;

00 O 0o 0NH-\ O NH NH O 0 NH or O HN 0

% O O ; ;

00

00 HN NH0HN NH NH 0 0N O ; O ;

O 0 0 O p __ NH / 0 HN--- NH N O O HN 0 0

O 0--/~~0 O \/ N -- 0--/ O O o O 0 HN 00 HN HN ; ; ;

NH NH \__N _ ~\/ 0 O- /- __\& _ / 0 O O

O HN 000 HN

O O NH NH O -- O % 0 0 HN-- O O O

o O 0360 NH O O O O O HN

Oct 2023

O 0 0 O O -- 0 \/- 0 OHN---- HN 0 0HN-- o O HN

0 0 O 0_C __- HN--- HN -NIH 2023248067 10 NH N N -- N N N- 0-- O N - O 0HN-- HN O

o N NH N NH 0 O 0 O ;

O o a HN HN--- 0 0 N NH N N N -- N o O 0 HO O 00 N O HN N NH

0 HN OHN-- N N 00 HN o HN N O 0 O N NH HN O N N N 0 00 N0 N N N N O NH O O NH O N N N- N O ;

0 0 --H-C - N N N O NH N NH N- // O -N N O 0-N- N -N-- N N' N 0 0N N N O N N NH N NH N HN O O ;

0H 0 N HN o -- CNN CNN /-' NH O HN NH%0HN o NH N0 ;/ C ; N0 NH O O O ;

362

0 0 0 0 2023248067 10 Oct 2023

O O O O NH NH NH NH 0 O HN; HN HN -- HN 0 o

N-O N-O O N-O O ,\~ N-0 O N-0N

0I N-ON- N 0 ,N N-O ;

N- N-0

O N-O O N-O O N.-a N-0 -a N-

o N-O N-O O N-0 0 O N- N

N-O N-O 0,, N-0 0 O N

0"- 7o~ N-O N-0N O N-O O

N-O O N-ON-0 N. a'o --- N-0 -a O

1N a - N-0 Na0 N-0 N-O N-O O O ;

"iao -0 N-O N-0 N-O

N- -C - 0-- N-- N-0 7 N-O N-O ;

1N ~ N-3 N-O N-O ;

N-O N-O N N ;

N N-o N~ N N N N -N-

N \NN N N N N-o N

2023248067

N N N-o N N 0 N -0

N --

N N N 1/ -0N N N N N

N N-fK'N--X\o N N-0 N N-O N N-O

(N- 'N N

f-\NjN N- N-O N N\-0) N-o N - -N N

0 0-- /-0 0- -- a 0--- a a __-- a- __*-/--a a-/ Ó

O '-- aN - ; 0 0-/b- -0 1 0_ 0- 0. 10. O

0I 0Q 00_ j-0

-- a- a.- aa--a-0 a-1 \ rl

0 ,-, a a a

-- a a --- '--a a0a-*--0\ 0-i 0--.

-- a a a O a--. -- a a0 a--; -- ao

-- a~__ a0-.- -- N /-\N-- --N N --.

364

Oct 2023 r\ 0- 0- -- N N-/ 0 -- N N-J 0-- -- N N- I---\ / d0 -- N N N N N N N N Ó N N

N N N--- N 00 0

2023248067 10 N N N N O N N -- N\ N-- -- - N- O N- ''C 0- N N-

O N N N N N N N \N--- -- N '- N N - \ N N N

N N N N N N- -- --N -\N-- - -- -N 0 0-

N N N N N N O -- N N-\-/ N 0 -- 0--- 0\ -- N /N --N N 0-- N N N N 0-N N 0 HN N N O

0_ HN 00N- 0 0 O NH O o NH O O O O NH -0o -- 0 0-\ \I H\ -- 0 0 HN--.- 0 0 0 O O O NH O HN 0 O O H O o0 0 0 ,,, 0H 0 -NH -0 0 o-' O ---.-- 0 0 O '' HN 0 O NH O o NH o o o O0 0NO o 0H--0 0 / NH -- 0 -- 0 NH o o O NH HN o O HN 0 0 HN 0 O O O H -- T NH0HNH 0~~ ~~ 00H~~ N0 o o --NH 0-- O O NH N O NH 0 -- 0 O o O O HN

o o N\H NH 0-- o NH NH 0 0NH O O NH 00a O O O

o o 0- o V0 NH NH 0 O NH NH lNH NH o 0 -- 0 0_/ 0 0 ; O O

365

O 0 ~ 00 O O NH -0 NH NH -- 0 __ 0 .- 0 0-- __- 0O O O O O 0 O 0O 0 O 0 O NH NH 0 O NH NH NH NH -- 0 'FA-0 '*--0 o -0O O O O O NH >NH -- O 0 0--/.--0 o 0- 0 O HN---. -- 0 HN O -0O -0 O N HN 0 O O O 0 O -- 0 HN--- 0 0 N O O HN O HN ;

O O

O O 00 o O HN O HN ;

0 o 0 O O O O o- HN O O a HN OHN; 0 HN- o O O 0 HN HN ;

O O o o a a HN-0- HN O O HN ;

O NH O o0 HN O N N o 00 HN O HN O

00 o 00_ -N O -- N-- NH NNH- o N N 0--/0 H N o 0-- H-- 0H N N N N NH o HN O 00 o o O O HN N N NH N N O

0 O 0 o O NH O -- N N/ \N- N 0-- '- -- N N N- N \ OHN-- O HN

366

000 O HN HN HN O O O O O N 0-- N -- N --N N- N \ j -0 O HN~; HN N N

0 O O N N~-- J NH NH N N N/\N---' -- N N HN __0 o-0N N N -O N NH 0 O ;

N NN \ N N N -- 0 0--- \s-j - NH--C/ C- NH o NH O O NH 00 O ; O ;

N/ N \N/N/ N N -- N --N N-- N - NH NH -- N --N N N NH NH 0C O O ;

N O --N N N -N N ~~N ' 0-N N N N NH NH 0 NN N HN 0 O ; O ;

N-0 0 N-O N-O N-0 N-O N- N-O O ;

N-O N-O N-O O O

N-O N-O N-O O 0 o 0 N N- N

N-O N-O N-O o

N-0 N-O N N-O O 0 N-0 N- N-O N-O O ;

0 0 N- N-O N- N-O I / -. I

367

N- 0 N- 0 10 Oct 2023

N-O N-O 0/ I / -

0 N- N-O N- 0 N-O O O

N- 0 N-O / \ N.- 0 N " N /---N O N N-O O N N -/N N 1 2023248067 0 O ; NN

N-O 0 N-0

[--N N- N N- N-O f("N N N N N-O N N-/-N~ N N

N-O N N-O N N-O N N O N N O 0 N-0 N N N- N-O r-,N N-O N N / N) N 74. N N-C0N- N-C N-O N-O N O N N N O

N-O NI/ N N --- N-N-N N N ; andN)/7 N ; and

108871 In any

[0887] In any aspect aspect or embodiment or embodiment described described herein, herein, the ALthe isA'is selected selected from: from:

O C 0 O ; O OH OH o 0

N.0~ 0 O 0 o o

o 00 O

368

>00O

0 %> a 0 IZ H 0 0 < 0 0

IZ H0 0 0 H N N N

O O K'Z 0 / 0*o

O 00

0 ~0

0 0

0 0

mm ~ N

0 0 mm

0 I N

JVVV N nn in N 0 0

369 69£

IZ IZ H N H H H ZI ZI H H IZ N IZ N N N H H O O

ZI ZI N H H 2023248067

O O IZ ZI IZ IZ N N N H ZI ZI

H H HH ZI IZ H H IZ N ZI N N~ /' O- N H H O O

IZ ZI N H H HH H O O ZI IZ IZ IZ N N H HH H HH H H

IZ ZI H H H H ZI H ZI N kN~N ZI

H H H H O IZ IZ IZ IZ N H H H HHH H

IZ IZ IZ IZ N N N H H H H I370 ZI H IZ o IZ IZ N N N N H H H F F F F F,F ZI H IZ IZ ZI N N H H H F F

HN NH X-X-O \/ X-Yi:NH -X-Y-: HN\ |---X-0

F - N N NH |---X-O HN \/HN NH \/ -X-Y----X-O

F EL

NH HN / 0~iHN |---X-0 \ X-Yi: NH -A-X-O N N

N NH X NH N N N -NN EL F ZI X ZI X ZI X N N N H H H O H H O H / F E

N IZ N X I-N/ ZI \- X ZI X -N HH O H H /0H O H

IZ N ZI N N HH /Y X HH N ' N N

N~ 0 IZ N N ZI N N N H N H

\ / N--\ N 0- \ N-\ O O IZ N N N N HN 0 o0 o

N N N N ,I:)"NH-X-Y -YX HN X " A-NH HN I \ -NH HN

wherein: wherein:

371

Oct 2023 'X" inin above 'X" abovestructures structurescancan be be linear linear chain chain withwith atomsatoms ranging ranging from 2 from to 14,2 and to the 14, and the mentionedchain mentioned chaincan cancontain containheteroatoms heteroatoms such such as as oxygen; oxygen; andand

"Y" in above "Y" in abovestructures structures can can be be O, 0, N, N, S(O) S(O),(n=0, (n=0,1,1,2). 2).

[0888]

[0888] In any aspect In any aspect or embodiment or embodiment described described herein, the the herein, linker (L)(L) linker comprises comprises a structure a structure 2023248067 10

selected selected from: from:

(YL¹)- yLl)0-2 (YL¹) (yLl)0-2 WL1 WL2 WL1 WL2 n n or or

wherein: wherein:

2 WL andWL² WL¹ and WLareareeach eachindependently absent, independently a 4-8 absent, a 4-8membered ring ring membered withwith 0-4 0-4 heteroatoms, heteroatoms, optionally optionally substituted substituted with with RQ, R°, each each RQ R° is independently aa H, is independently H, halo, OH, CN, halo, OH, CN,CF, CFC-C 3 , CI-C6

alkyl (linear, branched, alkyl (linear, branched,optionally optionally substituted), substituted), C 1-C 6 (linear, C-C alkoxy alkoxy branched, (linear, branched, optionally optionally

substituted), or22R°RQgroups substituted), or groups taken taken together together with with the thethey atom atomarethey are attached attached to, form ato, 4-8form a 4-8

membered ringsystem membered ring system containing containing 0-40-4 heteroatoms; heteroatoms;

YL¹ is each yLl is each independently independently aa bond, bond, C-C C -C alkyl alkyl 1 (linear,branched, (linear, 6 branched,optionally optionallysubstituted) substituted)and and optionally optionally one or more more CCatoms atomsare arereplaced replacedwith withO;0;ororC-C C1 -C6 alkoxy (linear, branched, alkoxy (linear, branched,

optionally substituted); optionally substituted);

n is n is 0-10; and 0-10; and

aa dashed line indicates dashed line indicates the theattachment attachment point point to tothe thePTM or CLM PTM or moieties. CLM moieties.

anyany

[0889] In In

[0889] or or aspect aspect embodiment embodiment described described herein, thethe herein, linkercomprises linker a structure comprisesa structure selected selected from: from:

(R°)- (R°)- Y(YLl)0-2 O W (YL¹)- L 1(yUl) Q wL 2 WL2 WL2 WL1) QL QL / n WL1 n n

or or

wherein: wherein:

WL and WL¹ and are2 are WL²WL each each absent, absent, independently independently aryl, heteroaryl, aryl, heteroaryl, cyclic, heterocyclic, cyclic, heterocyclic, C1-6 alkyl C -6 alkyl 1

and optionally one and optionally or more one or more CCatoms atomsare arereplaced replacedwith withO,0,C1-6 C 1-6alkene alkeneand andoptionally optionallyone one or more or more CCatoms atomsare arereplaced replacedwith withO,0,C1-6 C 1-6alkyne alkyneand andoptionally optionallyone oneorormore moreC C atoms atoms areare

372

2023 replacedwith replaced withO,0, bicyclic, bicyclic, biaryl, biaryl, biheteroaryl,or biheteroaryl,or biheterocyclic, biheterocyclic, each optionally each optionally

substituted substituted with with RQ, R°, each each RQ is independently R° is independently aa H, H, halo, halo, OH, CN,CF, OH, CN, CFhydroxyl, 3 , hydroxyl, nitro, C nitro, C

2023248067 10 Oct

- CH,C2-6 =CH, C2-6alkenyl, alkenyl, C2-6 C2-6 alkynyl, alkynyl, C-C 6 alkyl CI-Calkyl (linear, branched, (linear, branched,optionally optionallysubstituted), substituted), C 1-C 6 alkoxy (linear, branched, optionally substituted), OC1.3alkyl (optionally substituted C-C alkoxy (linear, branched, optionally substituted), OC1-alkyl (optionally substituted

by 11 or by or more -F), OH, more -F), OH,NH, NH NRY¹R², CN, 2or , NR'RY , CN, orgroups 2 R° 2 RQ groups taken together taken together with with the the atom atom 2

they are they are attached attached to, to,form form aa4-8 4-8membered ringsystem membered ring systemcontaining containing0-4 0-4heteroatoms; heteroatoms; Y" is YL¹ is each independently aa bond, each independently NRY",O,O,S,s,NRYL², bond, NRYL¹, NRYL 2CRYL¹RYL², , CRYLRYL 2 C=O, C=S, C=S, , C=, SO, SO, SO,C-S02, C1

C C 6alkyl (linear, branched, alkyl(linear, branched, optionally substituted)and optionally substituted) optionallyone and optionally ormore one or more C C atoms are atoms are

replaced with replaced with O; 0; C-C C1 -C6 alkoxy alkoxy (linear,branched, (linear, branched, optionally optionally substituted); substituted);

QL QL is a 3-6 membered alicyclic or aromatic ring with 0-4 heteroatoms, optionally bridged, is a 3-6 membered alicyclic or aromatic ring with 0-4 heteroatoms, optionally bridged, optionally optionally substituted substituted with with 0-6 0-6 RQ, R°, each each RQ R° is independently H, is independently H, C-C 1alkyl .6 alkyl(linear, (linear, branched, optionally branched, optionally substituted substituted by by 11 or or more halo, C more halo, C- .6alkoxyl), alkoxyl), oror 22 R° 1 RQgroups groupstaken taken together with together with the atom they are atom they are attached to, to,form form aa 3-8 3-8 membered ringsystem membered ring systemcontaining containing 0-2 heteroatoms); 0-2 heteroatoms);

2 R¹¹, , RYLeach RYL are areindependently each independently H, alkyl H, OH, C- OH, C(linear, 1 .6 alkyl (linear, branched, optionally substituted branched, optionally substituted

by 11 orormore by morehalo, C- C halo, alkoxyl), 1.6 or R¹, R² together 2 with the atom they are attached to, alkoxyl), or R, R together with the atom they are attached to, form form aa 3-8 3-8 membered membered ringsystem ring system containing containing 0-20-2 heteroatoms); heteroatoms);

n is n is 0-10; and 0-10; and

a dashed line indicates dashed line indicates the theattachment attachment point point to tothe thePTM or CLM PTM or moieties. CLM moieties.

[0890] In any

[0890] In aspect any aspect or embodiment or embodiment described described herein, herein, the the(L) linker linker is a (L) is a polyethylenoxy polyethylenoxy

group optionally group optionally substituted substituted withwith aryl aryl or phenyl or phenyl comprising comprising from 1 tofrom 1 to 10glycol 10 ethylene ethylene units.glycol units.

[0891] In any

[0891] In aspect any aspect or embodiment or embodiment described described herein,herein, the PTMtheisPTM is an estrogen an estrogen receptorreceptor (ER) (ER) binding moiety binding moietyrepresented representedbybythe the chemical chemicalstructure: structure:

XPTM1 XPTM1 XPM1-,\ RPTM5 RPTM 5 RPTM3-P ( RPTM3 RPTM3 XPTM2 RPTM3 XPTM2 XPTM2 RPTM4 RPTM4 XPTM XpTM XpTM XPTM

RPTM2 RP-- RPTM2 RPTM2 R RPTM1 SS S RPTM1 RPM1 S S

PTM-I PTM-I or or PTM-II PTM-II wherein: wherein:

373

C=O; or C=O; is O0or 2023248067 10 Oct 2023

XPTM XPTM is

eachof each of XPTM1 andXPTM2 XPTM1 and XPTM2 is isindependentlyselected independently fromN N selectedfrom or or CH; CH;

RPTM1 is independently RPTM1 is independentlyselected selectedfrom from OH,OH, O(CO)RPTM, O(CO)RPTM, O-lower O-lower alkyl, wherein alkyl, wherein RPTM is RPTMis an an alkyl or aryl alkyl or aryl group groupin inthetheester; ester; andRPTM4 RPTM2 and RPTM2 RPTM4 areindependently are independently selected selected from from H, halogen, H, OH, OH, halogen, CN, CN, CF, CF 3 , S02-alkyl, SO2-alkyl, O- 0 lower alkyl; lower alkyl;

andRPTM5 RPTM3 and RPTM3 areindependently RPTM5are independentlyselected selectedfrom fromH,H,halogen; halogen; PTM-I hasatatleast PTM-I has least one RPTM2and one RPTM2 at least and at least one one on each RPTM3 on RPTM3 eachrespective respective rings; rings; and and

the the indicates the site indicates the site of of attachment attachment ofofat at least least one oneofofthe thelinker, linker, the the CLM, CLM, a CLM', a CLM', or a or a

combination thereof. combination thereof.

[0892]

[0892] In In any aspect or any aspect or embodiment described embodiment described herein,thethePTM herein, PTM is an is an estrogen estrogen receptor receptor (ER) (ER)

binding moiety binding moietyrepresented representedbybythe thechemical chemicalstructure: structure:

RPTM4 XPTM XpTM XpTM XPTM RA RPTM1 RPTM1 XpTM X-PTM RPTM3 RPTM3 XpTM XpTM

RPTM2 RPTM2

or or

Formula(IPTM) Formula (IPTM)

RPTM1 RPTM1 XpTM XPTM

RPTM3 RPTM3 XPTM XpTM

RPTM2 RPTM2

374

Oct 2023 Formula Formula (IIPTM) () wherein: wherein:

each XPTMisisindependently each XPTM independentlyCH,CH, N; N;

2023248067 10

3 indicates the site aa VLM, MLM, VLM, MLM, site of

a ULM', a ULM', ofattachment

a ILM', a ILM', of at attachment of

a VLM', a VLM', at least leastone

a MLM', a MLM', one of

or a or of the the linker linker(L),

a combination combination (L),the

thereof; thereof; theCLM, CLM, aa CLM', CLM',ULM, ULM, an ILM, an ILM,

each each is independently RPTM1 is RPTM1 independently OH, OH,halogen, halogen,alkoxy, alkoxy,methoxy, methoxy, ethoxy, ethoxy, O(CO)RPTM, O(CO)RPTM, wherein wherein the the substitution can be aa mono-, can be mono-,di- di-orortri-substitution tri-substitution and the RPTM and the is alkyl RPTMis alkyl or cycloalkyl group or cycloalkyl groupwith with1 1 to 6 carbons to or aryl carbons or aryl groups; groups; each RPTM2isisindependently each RPTM2 independentlyH, H, halogen, halogen, CN, CN, CF 3, or CF, liner liner or branched branched alkyl, alkoxy, alkyl, alkoxy, methoxy,methoxy, ethoxy, ethoxy,

whereinthe wherein thesubstitution substitutioncan canbebemono- mono-or or di-substitution; di-substitution;

each each RPTM3 is independently RPTM3is independentlyH,H, halogen, halogen, wherein wherein the the substitution substitution can can be mono- be mono- or di-substitution; or di-substitution; and and

RPTM4 is a H, alkyl, methyl, ethyl. RPTM4is a H, alkyl, methyl, ethyl.

[0893] In any

[0893] In any aspect aspect or embodiment or embodiment described described herein, herein, theisPTM the PTM is an androgen an androgen receptorreceptor (AR) (AR) binding moiety binding moiety(ABM) (ABM) represented represented by by a structure a structure selectedfrom selected from thethe group group consisting consisting of:of:

Y² R¹ Y2R R2 R² R (R°)- (RQ)o-6 5 3Y W¹ N N Y³ Y W² N Y Q 4

Y¹ W² w¹

ABM-a ABM-a ; ABM-b ABM-b

Y¹1 Y Y 3 ~ 2 Y3 Ra Rª w2 W² _ (R²)- w¹ N w² Y³ Rb (Y³)-5 R Q Y Y² W W¹

ABM-c ABM-c ;and ; and ABM-d ABM-d

wherein: wherein:

W¹ 1isisaryl, W aryl, heteroaryl, heteroaryl,bicyclic, bicyclic,or or biheterocyclic, biheterocyclic, each each independently independently substituted substituted by 1 or by 1 or

more H,H,halo, more halo, hydroxyl, hydroxyl, nitro, nitro, CN, C=CH, CN, C=CH, C1 -6 C1-6 alkyl(linear, alkyl (linear, branched, branched, optionally optionally substituted; forexample, substituted; for example, optionally optionally substituted substituted bymore by 1 or 1 orhalo, moreC1-6 halo, C1-6 alkoxyl), alkoxyl), C- C1-6

alkoxyl(linear, alkoxyl (linear,branched, branched, optionally optionally substituted; substituted; for example, for example, optionally optionally substituted substituted by 1 by 1 or morehalo), or more halo),C2-6 C2-alkenyl, 6 alkenyl, C2-6 C2-6 alkynyl, alkynyl, or CF 3 ; or CF;

375

Oct 2023 Y, Y Y¹, Y²2 are are each each independently NRYO,1, O, independently NR¹, S, S, S02, SO2, heteroaryl, heteroaryl, or or aryl; aryl;

Y 3 ,Y Y 5are Y,4 , Y are each independentlya abond, each independently bond,0, O, 2 2 Y³, NRY , CRYIRv NRY², CRY¹R²,, C=, C=S, SO, C=0, C=S, SO, SO2, SO,

heteroaryl, ororaryl; heteroaryl, aryl; Q is aa 3-6 Q is 3-6 membered ringwith membered ring with0-4 0-4heteroatoms, heteroatoms,optionally optionallysubstituted substitutedwith with0-6 0-6R°, RQ,each each 2023248067 10

R,is R°, is independently independently H, H, C- alkyl (linear, branched, optionally substituted, for example, C 1.6 alkyl (linear, branched, optionally substituted, for example, optionally optionally substituted substituted by by 11 or ormore more halo, halo, CC-.6alkoxyl), 1 alkoxyl), halogen, halogen, C- C 1alkoxy, .6 alkoxy,oror2 2R°RQ groups taken together groups taken together with with the the atom atom they they are are attached attached to, to, form form a 3-8 3-8 membered ring membered ring

system containing 0-2 system containing 0-2 heteroatoms); heteroatoms); R', R¹, R 2 , R, R², R, R, R 1R² R,R¹¹, , RY2 areare each each independently independently H, alkyl H, C- C 1 .6 alkyl (linear, (linear, branched, branched, optionally optionally

substituted; for substituted; forexample, example, optionally optionally substituted substitutedby by1 1orormore more halo, halo,CC- 1 .6 alkoxyl), halogen, alkoxyl), halogen,

C 2 C- alkoxy, cyclic, heterocyclic or R¹, R² together with the atom they are attached to, 1 .6 alkoxy, cyclic, heterocyclic or R, R together with the atom they are attached to,

form form aa 3-8 3-8 membered membered ringsystem ring system containing containing 0-20-2 heteroatoms); heteroatoms);

W² 2isisa abond, W bond,C- Calkyl, C- heteroalkyl, O, aryl, heteroaryl, alicyclic, heterocyclic, 1 .6 alkyl, C 1 .6 heteroalkyl, 0, aryl, heteroaryl, alicyclic, heterocyclic,

biheterocyclic,biaryl, biheterocyclic, biaryl,ororbiheteroaryl, each optionally biheteroaryl,each substituted optionally by 1-10 substituted byR²; 1-10 Rw 2 each RW2 each RW² is isindependently independently H, halo, H, halo, C 1 .6(linear C- alkyl alkyl (linear or branched or branched optionally optionally substituted;substituted; for for example, optionally substituted substituted by 2 C- cycloalkyl, C4-6 example, optionally by 1 1 or more F), or more -ORW2A F), -ORw A , C3-6 cycloalkyl, C4 -6

cycloheteroalkyl, cycloheteroalkyl, C- C 1 6 alkyl (optionally substituted), , heterocyclic (optionally substituted), alkyl (optionally substituted), heterocyclic (optionally substituted),

aryl (optionally aryl (optionallysubstituted), substituted),or or heteroaryl heteroaryl (optionally (optionally substituted), substituted), bicyclic bicyclic hereoaryl hereoaryl or or aryl, OC1.3alkyl aryl, (optionallysubstituted; OC-alkyl (optionally substituted; for for example, optionally substituted example, optionally substituted by by 11 or more more

-F) , OH, -F), OH, NH 2 , NR NH, 'Rv 2, CN; NRY¹R², CN; RW2 AisisH,H,C-Calkyl R²²A 1 (linear, .6 alkyl branched), (linear, or C1-6orheteroalkyl branched), (linear,(linear, C .6 heteroalkyl 1 branched), each optionally branched), each optionally substituted bya acycloalkyl, substituted by cycloalkyl, cycloheteroalkyl, cycloheteroalkyl, aryl, aryl, heterocyclic, heterocyclic, heteroaryl, heteroaryl, halo, orhalo, OC1- or OC1

and 3alkyl; and alkyl; .

the dashed the dashedline lineindicates indicatesthethesite siteofofattachment attachmentofof atatleast leastone oneofofthe thelinker, linker, the the CLM, CLM, a CLM', a CLM', or aor a combinationthereof. combination thereof.

[0894] In In

[0894] anyany aspect aspect ororembodiment embodiment describedherein, described herein, the the PTM PTMisisa aBET/BRD4 BET/BRD4 targeting targeting

moiety comprising moiety comprisinga agroup groupaccording according to to thechemical the chemicalstructure structurePTM-a: PTM-a:

376

Oct 2023 Z,

A)Y Z A Y YJN N Y1

N Y Y O 0 3 B B N 2023248067 10

PTM-a PTM-a

wherein: wherein:

2 and Y 3 are independently selected from the group of carbon, nitrogen or oxygen Y 1,YYand Y, Y are independently selected from the group of carbon, nitrogen or oxygen and and with the atoms together with together to form atoms to aromatic fused an aromatic form an fused ring. ring. A and A andB Bareareindependently independently selected selected fromfrom the group the group of a 5-membered of a 5-membered aromatic aromatic ring, ring, a 6- a 6 membered membered aromatic aromatic ring,a heteroaromatic ring, a heteroaromatic ring,a acarbocyclic, ring, carbocyclic,a athiophene thiophenea apyrrole pyrrolering, ring, aa pyridine, pyridine, a apyrimidine, pyrimidine, a pyrazine, a pyrazine, a pyrazole a pyrazole ringoptionally ring each each optionally substituted substituted with alkyl, with alkyl, alkoxy, halogen, an alkoxy, halogen, an aromatic aromaticand anda heteroaromatic a heteroaromatic ring; ring; wherein wherein ringring A isAfused is fused to the to the

central central azepine azepine (Yl=C) ordiazepine (Y1=C) or diazepine(Y1 (Y1= =N)N)moiety; moiety; andand

Z Iis Z1 is selected selected from from the the group of methyl group of or analkyl methyl or analkyl group, group, and and whereinthethedashed wherein dashed lineline indicates indicates the site the site of attachment of attachment of atone of at least least of one of the the the linker, CLM,the linker, CLM, aa CLM', CLM', orora acombination combinationthereof. thereof.

[0895]

[0895] In In any aspect or any aspect or embodiment embodiment described described herein, herein, thethe PTMPTM is a is a BRaf BRaf targeting targeting moiety moiety

that that is is represented represented by by at at least leastone one of of chemical structures PTM-Ia, chemical structures PTM-Ib, PTM-Ia, PTM-Ib, PTM-IIa, PTM-IIa, PTM-IIb, PTM-Ilb,

PTM-IIIa, PTM-IIIb, PTM-IIIa, PTM-IIIb, PTM-IVa, PTM-IVb: PTM-IVa, PTM-IVb:

RPTM1 RPTM1

XpTM XPTM WPTMWPTM RPTM2 RPTM2YPTM-RPTM3 YPTM-RPTM3 VPTM ZPTM RPTM4 RPTM4 N,

N HO NN HO

PTM-Ia PTM-Ia

377

RPTM1 RPTM1

X)PT! XPTM RPTM2RT2 WPTM WPTM' I' PTM2 ,YPTM-RPTM3 YPTM-RPTM3 VPTM vV ZPTM ZPTM XPTM35 RPTM4 RPTM4

RRPTM5 RRPTM5 O H/ ZI 2023248067

H N S

O 0PM3, XPTM36 .XPTM38 XPTM38 N XPTM37 XPTM37

PTM-Ib PTM-Ib

RPTM5a RPTM6a RPTMa RPM~aRPTM8 RPTM8

0o RPTM7 RPTM7 XPTM3 XPTM3 RPi-MVI RPTM9 O RPTM6 XPT' RPTM6 XPTM2 ~XPTM4 XPTM4

RPTM6b XPTM1 I II XPTM6 XPTM5 RPTM10

NXPTM11 pT~ RPTM11 ZI N H N PTM-Ila PTM-IIa

RPTM6a RPTMa RPM~aRPTM8 RPTM5a RPTM8

0 ~RPTM7R RPTM7 PTM9 RPTM9 O RPTM6 N RPTM6 N

XPTM1 N RPTM~bI .1 RPTM10 RPTM6b

i RPTM11 ZI N H N

378

PTM-Ilb PTM-Ilb 0 XPTM16 XPTM16 O XPTM15 XPTM15 RPTM13 RPTM13

I PT14 RPTM14 \ PTM 10RPTM12 XPTM10 RPTM12 ZI RPM1HH IIIIIIIII XPTM17 XPTM17 0 o TR 17~i RPTM17 XPTM1< XPTM11 XPTM9 XPTM9 RPTM18 N PTM20 XPM4NPM

2023248067 XPTM20 / \N N XPTM1 XPTM12 XPTM14XPTM8 XPTM7 XPTM7 RPTM 16 RPTM16

XPTM19 XPTM19 R x1 XRPTM15 RPM9XPTM18 RPTM19 XPTM18 XPTM13 13 TM t R RPTM15

N N N IZ N 0 O H H RPTM20 RPTM21 RPTM21 RPTM20 ,

PTM-111 PTM-III

RPTM22 RPTM22 0 O RPTM25a RPTM6 N S NH RPTM25a RPTM26 0 RPTM25 N O RPTM25 XPTM29

XPTM2" XPTM21 XPTM~23 XPTM23

N / N~ K XPTM27 XPTM27 N XPTM28 N RPT2 XPTM22/ XPTM22 RPTM28

W ~ RPTM27 RPTM27 RT2 RPTM29 RPTM25b RPTM25b XPTM24 ,XPTM26 RPTM30 RPTM30 M2 XPTM26 XPTM24 '~ / XPTM33 XPTM 4 -XPTM33 XPTM34 XPTM254 XPTM25 \ . RPTM24 RPTM24 N XPTM32 XPTM32 N XPTM30 XPTM31 TM31 RPTM RPTM23 XPTM3O-

RPTM31 RPTM31 / RPTM32 RPTM32

PTM-IVa PTM-IVa

379

Oct 2023

RPTM22 0 O S NH RT2 NH RPTM26 NH RPTM25a RPTM25a RPTM26 N N 2023248067 10 O O RPTM25 RPTM25 XPTM29 x

XPTM21 XPTM21 XPTM23 XPTM23 XPTM27 XPTM27 / N N XPTM28 XPTM22 N XPTM28 RPTM28 RPTM28

RPTM27 RPTM27 RPTM29 RPMT25b RPMT25b XPTM24 XPTM26 RPTM30 RPTM30 RPTM29 XPTM26 XPTM24 XPTM34 XPTM33 XPTM3 4 -XPTM33 XPTM25 XPTM25 RPTM24 RPTM24 XPTM32 XPTM32 N N XPTM30 XPTM31 RPTM23 RPTM23 XPTM 3 0-XPTM31 RT RPTM32 RPTM31 RPTM31

PTM-IVb PTM-IVb wherein: wherein:

double dotted bonds double dotted bondsare are aromaric aromaricbonds; bonds; VPTM, VPTM, WPTM, WPTM,XPTM, YPTM, XPTM, ZPTM YPTM, is one iS ZPTM ofone the of the following following combinations: combinations: C, N, C, CH, CH,N,N,C; N, C, C; N, C, N, N, CH, N, CH,C;C;C,C,O,O,C,C,CH, CH, C; C; C, C, S, S, C, C, CH,CH, C; CH, C; C, C, CH, C, O,C,C; O,C,C;CH, C, C,CH, S, C, C; S, C, C; CH,C,N,CH, N, CH, C;N, CH, C; N, CH, CH,C,C,CH,CH, C; C; C, C, CH,CH, C, CH, C, CH, N; N,N;N,N,C,N,CH, C, C;CH, N, C; CH,N,C,CH, C, C, N, C; N, CH, C; C, C, CH, C,

N, N; N, C, N, C, N; C, C, CH, CH,N; N;C,C,N,N,C,C,N, N, C;C;and andC,C,N,N,N,N,N,N,C;C; RPTM1 covalently joined RPTM1 isiscovalently joined totoa ULM, ULM, a achemical chemical linkergroup linker group (L),a CLM, (L), a CLM, an ILM, an ILM, a VLM, a VLM,

MLM, MLM, a aULM', ULM',a aCLM', CLM',a aILM', ILM',a aVLM', VLM',a aMLM', MLM',or or combinationthereof; combination thereof; RPTM2 is hydrogen, RPTM2 is hydrogen,halogen, halogen,aryl, methyl, aryl, ethyl, ethyl, methyl, OCH 3 , OCH, NHCH 3 or or NHCH M1-CH 2-CH 2-M2, M1-CH-CH-M2, wherein M1 wherein M1isis CH, CHO2 , and 0 and NH, NH, and M2and is M2 is hydrogen, hydrogen, alkyl, alkyl, cycliccyclic alkyl,alkyl, aryl aryl or or heterocycle; heterocycle;

RPTM3 isis absent, RPTM3 absent, hydrogen, hydrogen,aryl, aryl, methyl, methyl,ethyl, ethyl, other other alkyl, alkyl, cyclic cyclic alkyl, alkyl, OCH, OCHNHCH 3 , NHCH or 3 or M1-CH 2 -CH 2-M2, M1-CH-CH-M2, wherein wherein M1 isM1 CH,isOCH 0 and 2 , NH, and andNH, andhydrogen, M2 is M2 is hydrogen, alkyl, alkyl, cyclic cyclic alkyl, aryl or alkyl, aryl or heterocycle; heterocycle;

380 is hydrogen, hydrogen,halogen, 2023248067 10 Oct 2023

RPTM4 RPTM4 is halogen,aryl, methyl, aryl, ethyl, ethyl, methyl, OCH 3 , OCH, NHCH 3 or or NHCH M1-CH 2-CH 2-M2, M1-CH-CH-M2, wherein M1isis CH, wherein M1 CHO2 , and 0 and NH, NH, and and M2 is M2 is hydrogen, hydrogen, alkyl, alkyl, alkyl,alkyl, cycliccyclic aryl aryl or or

heterocycle; heterocycle;

each of RPTM5 each of andRPTM22 RPTM5 and RPTM22is isindependentlyselected independently selectedfrom fromthe groupconsisting thegroup consistingofof F F F F

N----- N N--- N F F N-- N ---------- N-- N N N ----- ----- ----- N----- F F ---- N --- FF N----- FF --- F F N-- N N F F F F HO N ----- : N ---- N ----- HO O* -N N_ N HOIIIIII.

HO HO HO HO-- Ho

HO Ho ;

XPTM1, XPTM1, XPTM2, XPTM2, XPTM3, XPTM3,XPTM4, XPTM4,XPTM5, XPTM5,XPTM6, XPTM7, XPTM6, XPTM8, XPTM7, XPTM9, XPTM8, XPTM10, XPTM10, XPTM9, XPTM11, XPTM11, XPTM12, XPTM12,

XPTM13, XPTM14, XPTM15, XPTM13, XPTM14, XPTM15, XPTM16, XPTM17, XPTM18, XPTM16, XPTM17, XPTM18,XPTM19, XPTM19,XPTM20, XPTM21,XPTM22, XPTM20,XPTM21, XPTM22,XPTM23, XPTM23,

XPTM24, XPTM24, XPTM25, XPTM25, XPTM26, XPTM26, XPTM27, XPTM27, XPTM28, XPTM28, XPTM29, XPTM29,XPTM30, XPTM31,XPTM32, XPTM30,XPTM31, XPTM32,XPTM33, XPTM33,XPTM34, XPTM34,

XPTM35, XPTM35, XPTM36, XPTM36, XPTM37, XPTM37, XPTM38 are independently XPTM38are or or fromCHCH selectedfrom independentlyselected N; N;

RPTM5a RPTM5a is is selectedfrom selected from the the group group consisting consisting of: H, of: H, optionally optionally substituted substituted amide amide (e.g., (e.g., optionally substituted optionally substituted with with an alkyl, an alkyl, methyl, methyl, ethyl,ethyl, propyl, propyl, or group), or butyl butyl group), optionally optionally

RPTM5 0 o S S { NH NH o \- substituted substituted amine, o amine, -NHC(O)RPTM5; , -NHC(O)RPTM5;

381 areeach independently eachindependently selected from hydrogen, or C-Coralkyl halogen, alkyl 2023248067 10 2023

RPTM6a RPTM6a andRPTM6b and RPTM6bare selected from hydrogen, halogen, Cl-C6

(linear, (linear, branched, optionally branched, optionally substituted); substituted);

Oct RPTM6 is either RPTM6 is either of of the thefollowing following groups: groups: absent, absent,hydrogen, hydrogen, halogen, halogen, aryl, aryl, methyl, methyl, ethyl, ethyl,OCH OCH,3

, NHCH NHCH or3 or M1-CH 2-CHwherein M1-CH-CH-M2, 2 -M2, wherein M1 is M1 is CH, O CH and2 ,NH, 0 and andNH, andhydrogen, M2 is M2 is hydrogen, alkyl,alkyl,

cyclic alkyl, aryl cyclic alkyl, aryl ororheterocycle. heterocycle. RPTM7 is isabsent, absent,hydrogen, halogen, hydrogen, aryl,aryl, halogen, methyl, ethyl, ethyl, methyl, OCH 3 , NHCH 3 or M1-CH OCH, NHCH 2-CH 2-M2, or M1-CH-CH-M2,

wherein M1 wherein M1isis CH, CHO2 , and 0 and NH, NH, and M2and is M2 is hydrogen, hydrogen, alkyl, alkyl, cycliccyclic alkyl,alkyl, aryl aryl or or heterocycle. heterocycle.

RPTM8, RPTM9oror RPTM8, RPTM9 RPTM10 RPTMoare are independently independently selected selected fromgroup from the the consisting group consisting of of absent, absent, hydrogen, halogen, hydrogen, halogen,aryl, aryl, heteroaryl, heteroaryl, alkyl, alkyl, cycloalkyl, cycloalkyl, heterocycle, heterocycle, methyl, methyl, ethyl, ethyl, OCH OCH, 3

, NHCH NHCH or3 or M1-CH 2-CHwherein M1-CH-CH-M2, 2 -M2, wherein M1 is M1 is CH, O CH and2 ,NH, 0 and andNH, andhydrogen, M2 is M2 is hydrogen, alkyl, alkyl,

cyclic alkyl, aryl cyclic alkyl, aryl ororheterocycle; heterocycle; RPTMll is RPTM11 is absent, absent,hydrogen, halogen, hydrogen, methyl, halogen, ethyl, methyl, 3 , NHCH OCHOCH, ethyl, or M1-CH NHCH3 or 2 -CH 2in M1-CH-CH-M2 -M2 in which M1, which M1, wherein wherein CH, CH O2 , and 0 and NH,NH, and and M2hydrogen, M2 is is hydrogen, alkyl, alkyl, cyclicalkyl, cyclic alkyl, aryl aryl or or heterocycle; heterocycle;

RPTM12, RPTM12, RPTM13, RPTM13, RPTM14, RPTM14, RPTM15, RPTM16,RPTM17, RPTM15,RPTM16, RPTM17, RPTM18, RPTM18, RPTM19 are are RPTM19 independently independently selected selected

from the group from the groupconsisting consisting of of absent, absent, hydrogen, hydrogen, halogen, halogen, aryl, aryl, heteroaryl, heteroaryl, cycloalkyl, cycloalkyl,

heterocycle, methyl, heterocycle, methyl,ethyl, otherother ethyl, alkyl,alkyl, OCH 3 ,OCH, NHCH 3 oror NHCH M1-CH 2-CH 2 -M2, M1-CH-CH-M2, wherein wherein M1M1

is is CH CH,2 ,00and andNH, NH,andand M2 M2 is hydrogen, is hydrogen, alkyl, alkyl, cyclic cyclic alkyl,aryl alkyl, arylororheterocycle; heterocycle; RPTM20 isisa small RPTM20 small group group containing less less than than four four non-hydrogen atoms; non-hydrogen atoms;

RPTM21 is selectedfrom RPTM21is selected from the the group group consisting consisting of trifluoromethyl, of trifluoromethyl, chloro,fluoro, chloro, bromo, bromo, fluoro, methyl, methyl, ethyl, propyl, ethyl, propyl,isopropyl, isopropyl, tert-butyl, tert-butyl, butyl, butyl, iso-butyl, iso-butyl, cyclopropyl, cyclopropyl, cyclobutyl, cyclobutyl, cyclopentyl, cyclopentyl,

cyclohexyl, OCH cyclohexyl, OCH,3 , NHCH, NHCHdimethylamino 3 , dimethylamino or M1-CH 2-CHwherein or M1-CH-CH-M2, 2 -M2, wherein M1 Ois M1 is CH, CH2 , 0 or NH,andand or NH, M2 M2 is hydrogen, is hydrogen, alkyl, alkyl, cyclic cyclic alkyl, alkyl, aryl oraryl or heterocycle; heterocycle;

RPTM25a RPTM25a and RPTM25b and are each RPTM25b are each independently selectedfrom independentlyselected hydrogen,halogen, fromhydrogen, or or halogen, Cl-C 6 alkyl C-Calkyl

(linear, (linear, branched, optionally branched, optionally substituted); substituted);

RPTM23, RPTM24, RPTM24, RPTM28, RPTM29, RPTM30, 30, RPTM31, RPTM32 are RRPTM31, RPTM32 independently selected are independently fromthe selected from the group consisting group consisting of of absent, absent, bond, bond, hydrogen, hydrogen, halogen, halogen, aryl (optionally aryl (optionally substituted), substituted),

heteroaryl(optionally heteroaryl (optionally substituted), substituted), cycloalkyl cycloalkyl (optionally (optionally substituted), substituted), heterocycle heterocycle

(optionally substituted),methyl, (optionally substituted), methyl, ethyl ethyl (optionally (optionally substituted), substituted), other other alkyl (linear, alkyl (linear,

branched, optionally branched, optionallysubstituted), 3 , NHCH OCHOCH, substituted), NHCH3 or orM1-CH 2 -CH 2-M2, M1-CH-CH-M2, wherein wherein M1 M1 is is

382

CH CH, 2O, 0 and NH, and and 2023248067 10 Oct 2023

and NH, M2hydrogen, M2 is is hydrogen, alkylalkyl (linear, (linear, branched, branched, optionally optionally substituted), substituted),

cyclic alkyl(optionally cyclic alkyl (optionallysubstituted), substituted), aryl aryl (optionally (optionally substituted)or substituted)or heterocycle heterocycle

substituted); (optionallysubstituted); (optionally

RPTM25 isisselected RPTM25 selected from from absent, absent, hydrogen, halogen, C-C hydrogen, halogen, Cl-C 6 alkyl alkyl (linear,branched, (linear, branched,optionally optionally substituted), substituted), OCH 3 , NHCH OCH, 3 orSCH; NHCH or SCH 3 ; RPTM26 isisselected RPTM26 selected from from absent, absent, hydrogen, halogen, C-C hydrogen, halogen, Cl-C 6 alkyl alkyl (linear,branched, (linear, branched,optionally optionally substituted), substituted), OCH3, OCH3,NHCH or SCH; NHCH 3or SCH 3; RPTM27 isisselected RPTM27 selected from from the the group consisting of group consisting of absent, absent, hydrogen, halogen, C-C hydrogen, halogen, 6 alkyl (linear, Cl-Calkyl (linear,

branched, optionally branched, optionallysubstituted), OCHOCH, substituted), 3 , NHCH NHCH3 or or SCH 3 ; and SCH; and at least at leastone one of ofRPTM8, RPTM8, RPTM9 or RPTM, RPTM9 or RPTM12, RPTM10, RPTM13,RPTM13, RPTM12, RPTM16, RPTM24, RPTM16, and RPTM32 RPTM24, RPTM29, is RPTM29, and RPTM32 is

modified to modified to be be covalently covalently joined joined to to aa ULM, ULM, a achemical chemicallinker linkergroup group(L), (L),a aCLM, CLM,an an ILM, ILM,

aa VLM, MLM, VLM, MLM, a ULM', a ULM', a CLM', a CLM', a ILM', a ILM', a aVLM', VLM', a MLM', a MLM', or or combination combination thereof. thereof.

[0896] any aspect In aspect

[0896] In any or embodiment or embodiment described herein, herein, described when when RPTM9 is RPTM9 is the covalently the covalently joined joined position, RPTM7 position, and RPTM8 RPTM7 and are connected RPTM8 are connectedtogether togethervia viaaa covalent covalent bond bondininaa way waytotoform forma abicyclic bicyclic group with the group with the ring ring to to which which RPTM7 andRPTM8 RPTM7 and areattached. RPTM8are attached.

[0897] any aspect In aspect

[0897] In any or embodiment or embodiment described herein, herein, described when when RPTM8 is RPTM8 is the covalently the covalently joined joined position, RPTM9 position, andRPTM10 RPTM9 and areare RPTM1o connected connected together together via avia a covalent covalent bond bond in in to a way a way to form a form a bicyclic group bicyclic with the group with the ring ring to to which which RPTM9 and RPTM10 RPTM9 and are attached. RPTM1oare attached.

[0898] In In

[0898] anyany aspect aspect or or embodiment embodiment described described herein, herein, when when RPTM10 RPTM10 is theiscovalently the covalently joined position, joined position, RPTM8 RPTM8 and and RPTM9 RPTM9 are connected are connected togethertogether via a covalent via a covalent bond in abond in way to a way to form form aa bicyclic bicyclic group with the group with the ring ring to to which which RPTM8 RPTM8 andand RPTM9 RPTM9 are attached. are attached.

[0899] In any

[0899] any aspect In aspect or embodiment or embodiment described herein,herein, described when is when RPTM12 RPTM12 is the covalently the covalently joined joined position, RPTM13 position, andRPTM14 RPTM13 and RPTM14areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form to a form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM14 RPTM13and which RPTM13 RPTM14areareattached, attached,and/or and/orRPTM15 andand RPTM15 RPTM16 RPTM16

are are connected togethervia connected together viaaa covalent covalentbond bondin ina way a way to to form form a bicyclic a bicyclic group group withwith the ring the ring to to

which RPTM15 which andRPTM16 RPTM15and areattached. RPTM16are attached.

[0900] In any

[0900] any aspect In aspect or embodiment or embodiment described herein,herein, described when is when RPTM13 RPTM13 is the covalently the covalently joined joined position, RPTM12 position, andRPTM16 RPTM12 and RPTM16areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form to a form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM16 RPTM12and which RPTM12 RPTM16areareattached, attached,and/or and/orRPTM15 andand RPTM15 RPTM16 RPTM16

are are connected togethervia connected together via aa covalent covalentbond bondin ina way a way to to form form a bicyclic a bicyclic group group withwith the ring the ring to to

which RPTM15 which andRPTM16 RPTM15and areattached. RPTM16are attached.

383 any aspect In aspect or embodiment herein,herein, described when is is the covalently joined 2023248067 10 Oct 2023

[0901] In any

[0901] or embodiment described when RPTM16 RPTM16 the covalently joined

position, RPTM12 position, andRPTM13 RPTM12 and RPTM13areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form toa form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM13 RPTM12and which RPTM12 RPTM13areareattached, attached,and/or and/orRPTM13 andand RPTM13 RPTM14 RPTM14

are are connected togethervia connected together via aa covalent covalentbond bondin ina way a way to to form form a bicyclic a bicyclic group group withwith the ring the ring to to

which andRPTM14 RPTM13and which RPTM13 RPTM14are attached. are attached.

[0902] In any

[0902] any aspect In aspect or embodiment or embodiment described herein,herein, described when is when RPTM24 is the covalently the covalently RPTM24 joined joined position, RPTM31 position, andRPTM32 RPTM31 and RPTM32areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form to a form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM32 RPTM31and which RPTM31 areare RPTM32 attached,ororRPTM29 attached, andand RPTM29 RPTM30 are are RPTM3o

connected together via connected together via aa covalent covalent bond bondininaa way waytotoform forma abicyclic bicyclicgroup groupwith withthethering ringtotowhich which RPTM29 andRPTM30 RPTM29 and RPTM3oare attached. areattached.

[0903] In any

[0903] any aspect In aspect or embodiment or embodiment described herein,herein, described when is when RPTM29 is the covalently the covalently RPTM29 joined joined position, RPTM24 position, andRPTM32 RPTM24 and RPTM32areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form toa form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM32 RPTM24and which RPTM24 RPTM32 areattached, are attached,and/or and/orRPTM31 RPTM31andand RPTM32 RPTM32

are are connected togethervia connected together via aa covalent covalentbond bondin ina way a way to to form form a bicyclic a bicyclic group group withwith the ring the ring to to

which RPTM31 which andRPTM32 RPTM31and RPTM32are attached. are attached.

[0904] In any

[0904] any aspect In aspect or embodiment or embodiment described herein,herein, described when is when RPTM32 is the covalently the covalently RPTM32 joined joined position, RPTM24 position, andRPTM29 RPTM24 and RPTM29areare connected connected together together via via a covalent a covalent bond bond in atoway in a way form to a form a bicyclic group bicyclic with the group with the ring ring to to which andRPTM29 RPTM24and which RPTM24 RPTM29 areattached, are attached,and/or and/orRPTM29 andand RPTM29 RPTM30 RPTM30

are are connected togethervia connected together via aa covalent covalentbond bondin ina way a way to to form form a bicyclic a bicyclic group group with with the ring the ring to to

which RPTM29 which andRPTM30 RPTM29and RPTM3o areattached. are attached. In any

[0905] In any

[0905] aspect aspect or embodiment or embodiment described described herein, herein, the PTM PTM thehas has a structure a structure selected selected from from the group consisting of: the group consisting of:

384

Oct 2023

R-s N N X = CI,Br, F,H, X = CI, Br, F, H,

SRS R S N NI bond, or aachemical bond,or chemical S N Y0 N moietycoupling couplingthe the O moiety NIN R the PTM CLMtoto the N / CLM PTM R N NN N 2023248067 10 11111 R N CI C10 xX R R S N NN N S N N R 7 N~ N N \/ O R Linker CI, Br, XX= CI, F, H Br, F, H J-NN - N N Likr NN R N R R N0 x O X N N -N 0 o S 'N N00% CONH O CONH2 " LNke Linker - 'NN X= CI, X Br, F, CI, Br, F, H Linker-N H Linker N -NN X CI, Br, F, H X = CI, Br, F, H

N N

x X x X

N N -NN S o 00 o X N o H LinkerNN Linker -NN X=CI, Br, F, H X = CI, Br, F, H NHN N H N

x X Linker Linker

385

Oct 2023

NN0 N S X N N o Z N 7 N N N / N N H H N 2023248067 10

""' N HN HN N Linker Linker c CI X ==H,IF X H, F O

R R R RX- XCI,Br, F, H, N NN0 X = CI, Br, F, H, bond, or achemical R N N~N 0 bond, or a chemical moiety coupling the R R O moiety coupling the

' CLM the PTMNN CLMtotothe PTM N N /

N N NH RN R N HN-R R I iN N N HN x X R R 0 O R R HO- Ho O-N O N I HO HO O - N - N N 00 O O R R

386

R Linker Linker R \N ~ N" N N N NNI O R IIIIIIIIII o N N N NH N N N NH NH

N N N/ N N N N 0 00

N, N 0 O IIIIIIIIII Linker Linker NH 2 o NH N\ N N NN N 0 -Linker Linker N~ N

N' N N N /

0

0 Linker\NR R Linker, o N NH2 N Z-O O NH N N- F F/\N\ N N -RR F-, 0 O N N F LinkerF Linker / \ NN. N N -o H0 H O

387

N N N N ~~--Linker Linker

F N N N 0 'N N-R N N N N / - Linker N-RN N F LLinker N R N N N N N Linker Lne H 0 H O , 2023248067

/ R R N Zzz NN /N N HN o N~~~N N N ,-N N /N - 0 N'~ N L~i~ N N~ Linker Linker NNN N N Lne Linker Linker Linker N N NH Lne NH

O o

388

N NN0 N H HN N N N CF 300 CF o O ANN 0 o S N-Linker N Linker

Linker O N H N LLinker 2023248067 O 0 N Linker HO F N F HN S N HO N N Li ne N ZI N N H O F F 0 -F N F NH FLinker F F N Linker F ZI N F H

FF F O N ZI H LiLinker N NH S F N

HN

O Linker

389

N N Linker Linker N N O===O - 0 O S/NH NH F N N N F F N N 0 N- Linker Linker F N O Nti N N OHN OZ|N / F F / N F F N O=S N IZ N N H N

,Linker Linker N 0 O O0ZI H N O\/ N S N /N -N V-NH F F N S N N N N Linker Linker N N N F F N N 'N F F F F O O N S 0 0 N-S NH N F NH F F O O N Linker NZ C'Linker O O N N -Linker Linker C C C F /F -N N F F N IZ N N IZ N H N N H wherein: wherein:

R is H, R is H, aa lower lower alkyl, alkyl, aabond, bond,or ora achemical chemicalmoiety moiety coupling the CLM coupling the CLM totothe thePTM; PTM;andand

Linker is Linker is aa bond or aa chemical bond or chemicallinker linkermoiety moietycoupling coupling thethe CLMCLM to PTM, to the the PTM, including including

pharmaceutically acceptable pharmaceutically acceptablesalt salt forms forms thereof. thereof.

[0906] In In

[0906] anyany aspector orembodiment aspect embodiment described compoundisisselected the compound herein, the describedherein, selected from the from the group consisting of group consisting of compounds 1-52. compounds 1-52.

[0907]

[0907] A further A further aspect aspectofofthe thepresent presentdisclosure disclosureprovides provides a composition a composition comprising comprising an an effective effective amount ofaa bifunctional amount of bifunctional compound compound of the of the present present disclosure, disclosure, andand a pharmaceutically a pharmaceutically

acceptable carrier. acceptable carrier.

390 or embodiment herein,herein, described further further the composition comprises at 2023248067 10 Oct 2023

[0908] In any

[0908] any aspect In aspect or embodiment described the composition comprises at

least one ofofadditional least one additionalbioactive bioactiveagent agent or another or another bifunctional bifunctional compound compound of the present of the present

disclosure. disclosure.

[0909] any aspect In aspect

[0909] In any or embodiment or embodiment described herein,herein, described the additional the additional bioactive agent agent bioactive is anti is anti-

cancer agent,anananti-neurodegenerative cancer agent, anti-neurodegenerative agent, agent, an antimicrobial an antimicrobial agent, an agent, an agent, antiviral antiviral agent, an anti an anti-

HIVagent, HIV agent, oror an an antifungal antifungal agent. agent.

[0910] An additiona

[0910] An additiona aspect aspect of theofpresent the present disclosure disclosure provides provides a composition a composition comprising comprising an an effective effective amount amount ofofatatleast least one onecompound compound of present of the the present disclosure disclosure and a and a pharmaceutically pharmaceutically

acceptable carrier,additive, acceptable carrier, additive, and/or and/or excipient excipient for treating for treating a disease a disease or disorder or disorder in athesubject, the in a subject,

methodcomprising method comprising administering administering the composition the composition to a subject to a subject in needinthereof, need thereof, whereinwherein the the compound is effective compound is effective in treating in treating or ameliorating or ameliorating at one at least least one symptom symptom of the of the disease disease or disorder. or disorder.

[0911] any aspect In aspect

[0911] In any or embodiment or embodiment described herein, herein, described the disease the disease or disorder or disorder is associated is associated

with theaccumulation with the accumulation and/or and/or aggregation aggregation of the protein. of the target target protein.

[0912] In any

[0912] any aspect In aspect or embodiment or embodiment described described herein, herein, the disease disease the or or is disorder disorder is selected selected from the group from the groupconsisting consistingofofasthma, asthma,autoimmune autoimmune diseases diseases such such as multiple as multiple sclerosis, sclerosis, various various

cancers, ciliopathies, cleft cancers, ciliopathies, cleft palate, palate, diabetes, diabetes, heart heart disease, disease, hypertension, inflammatorybowel hypertension, inflammatory bowel disease, mentalretardation, disease, mental retardation, mood mood disorder, disorder, obesity, obesity, refractive refractive error, error, infertility,Angelman infertility, Angelman syndrome, Canavan syndrome, Canavan disease, disease, Coeliac Coeliac disease, disease, Charcot-Marie-Tooth Charcot-Marie-Tooth disease, disease, Cystic fibrosis, Cystic fibrosis,

Duchenne muscular Duchenne musculardystrophy, dystrophy,Haemochromatosis, Haemochromatosis, Haemophilia, Haemophilia, Klinefelter's Klinefelter's syndrome, syndrome,

Neurofibromatosis,Phenylketonuria, Neurofibromatosis, Phenylketonuria,Polycystic Polycystic kidney kidney disease, disease, (PKD1) (PKD1) or 4 (PKD2) or 4 (PKD2) Prader- Prader

Willi syndrome, Willi Sickle-cell disease, syndrome, Sickle-cell disease, Tay-Sachs disease, Turner Tay-Sachs disease, Turnersyndrome. syndrome.

[0913] In any

[0913] any aspect In aspect or embodiment or embodiment described described herein, herein, the the or disease disease or is disorder disorder is selected selected from the group from the groupconsisting consistingofofAlzheimer's Alzheimer'sdisease, disease,Amyotrophic Amyotrophic lateral lateral sclerosis sclerosis (Lou (Lou Gehrig's Gehrig's

disease), Anorexia disease), nervosa,Anxiety Anorexia nervosa, Anxiety disorder, disorder, Atherosclerosis, Atherosclerosis, Attention Attention deficit deficit hyperactivity hyperactivity

disorder, Autism, disorder, Bipolardisorder, Autism, Bipolar disorder, Chronic Chronicfatigue fatiguesyndrome, syndrome, Chronic Chronic obstructive obstructive pulmonary pulmonary

disease, Crohn's disease, Crohn's disease, disease, Coronary heart disease, Coronary heart disease, Dementia, Depression, Diabetes Dementia, Depression, Diabetesmellitus mellitustype type1,1, Diabetes mellitus Diabetes mellitus type type 2, 2, Epilepsy, Epilepsy, Guillain-Barré Guillain-Barr6 syndrome, syndrome,Irritable Irritablebowel bowelsyndrome, syndrome, Lupus, Lupus,

Metabolic syndrome,Multiple Metabolic syndrome, Multiple sclerosis,Myocardial sclerosis, Myocardial infarction,Obesity, infarction, Obesity,Obsessive-compulsive Obsessive-compulsive disorder, Panic disorder, Panic disorder, disorder,Parkinson's Parkinson'sdisease, disease, Psoriasis, Psoriasis, Rheumatoid Rheumatoid arthritis, arthritis, Sarcoidosis, Sarcoidosis,

Schizophrenia, Stroke, Schizophrenia, Stroke, Thromboangiitis Thromboangiitisobliterans, obliterans, Tourette Tourettesyndrome, syndrome,Vasculitis. Vasculitis.

391

Oct 2023

[0914] any aspect In aspect

[0914] In any or embodiment or embodiment described described herein, herein, the disease disease the or disorder disorder or is is selected selected from the group from the groupconsisting consisting of of aceruloplasminemia, aceruloplasminemia, Achondrogenesis Achondrogenesis type II,type II, achondroplasia, achondroplasia,

Acrocephaly, Gaucher Acrocephaly, Gaucherdisease diseasetype type2, 2, acute acute intermittentporphyria, intermittent porphyria,Canavan Canavan disease, disease,

Adenomatous Adenomatous Polyposis Polyposis Coli, Coli, ALA ALA dehydratase dehydratase deficiency, deficiency, adenylosuccinate adenylosuccinate lyase deficiency, lyase deficiency, 2023248067 10

Adrenogenital syndrome, Adrenoleukodystrophy, Adrenogenital syndrome, Adrenoleukodystrophy,ALA-D ALA-D porphyria, porphyria, ALA dehydratase ALA dehydratase

deficiency, Alkaptonuria, Alexander deficiency, Alkaptonuria, Alexander disease, disease, Alkaptonuric Alkaptonuric ochronosis, ochronosis, alpha 1-antitrypsin alpha 1-antitrypsin

deficiency, alpha-i proteinase deficiency, alpha-1 proteinaseinhibitor, inhibitor, emphysema, emphysema, amyotrophic amyotrophic laterallateral sclerosis sclerosis Alstr6m Alström

syndrome, Alexanderdisease, syndrome, Alexander disease,Amelogenesis Amelogenesis imperfecta, imperfecta, ALA ALA dehydratase dehydratase deficiency, deficiency,

Anderson-Fabry disease, Anderson-Fabry disease, androgen insensitivity syndrome, androgen insensitivity syndrome,Anemia Anemia Angiokeratoma Corporis Angiokeratoma Corporis

Diffusum, Angiomatosis Diffusum, Angiomatosis retinae(von retinae (von Hippel-Lindau Hippel-Lindau disease) disease) ApertApert syndrome, syndrome, Arachnodactyly Arachnodactyly

(Marfan syndrome), (Marfan syndrome), Stickler Stickler syndrome, syndrome, Arthrochalasis Arthrochalasis multiplex multiplex congenital congenital (Ehlers-Danlos (Ehlers-Danlos

syndrome#arthrochalasia type) ataxia syndrome#arthrochalasia type) ataxia telangiectasia, telangiectasia, Rett Rettsyndrome, syndrome, primary pulmonary primary pulmonary

hypertension, Sandhoff hypertension, Sandhoff disease, disease, neurofibromatosis neurofibromatosis typeBeare-Stevenson type II, II, Beare-Stevenson cutis cutis gyrata gyrata syndrome, Mediterranean syndrome, Mediterranean fever, fever, familial, familial, Benjamin Benjamin syndrome, syndrome, beta-thalassemia, beta-thalassemia, Bilateral Bilateral

Acoustic Neurofibromatosis Acoustic Neurofibromatosis(neurofibromatosis (neurofibromatosis type type II),II), factorV V factor Leiden Leiden thrombophilia, thrombophilia, Bloch Bloch-

Sulzberger syndrome Sulzberger syndrome(incontinentia (incontinentiapigmenti), pigmenti),Bloom Bloom syndrome, syndrome, X-linked X-linked sideroblastic sideroblastic anemia, anemia,

Bonnevie-Ullrichsyndrome Bonnevie-Ullrich syndrome (Turner (Turner syndrome), syndrome), Bourneville Bourneville disease disease (tuberous (tuberous sclerosis), sclerosis), prionprion

disease, Birt-Hogg-Dub6 disease, Birt-Hogg-Dubé syndrome, syndrome, Brittle Brittle bone disease bone disease (osteogenesis (osteogenesis imperfecta), imperfecta), Broad Broad Thumb-Hallux syndrome Thumb-Hallux syndrome(Rubinstein-Taybi (Rubinstein-Taybisyndrome), syndrome),Bronze Bronze Diabetes/Bronzed Diabetes/Bronzed Cirrhosis Cirrhosis

(hemochromatosis), Bulbospinal (hemochromatosis), Bulbospinal muscular muscular atrophy atrophy (Kennedy's (Kennedy's disease), disease), Burger-Grutz Burger-Grutz syndrome syndrome

(lipoprotein (lipoprotein lipase lipase deficiency), deficiency), CGD Chronic CGD Chronic granulomatous granulomatous disorder, disorder, Campomelic Campomelic dysplasia, dysplasia,

biotinidase deficiency, biotinidase deficiency, Cardiomyopathy Cardiomyopathy (Noonan (Noonan syndrome), syndrome), Cri du Cri duCAVD chat, chat,(congenital CAVD (congenital absence of the absence of the vas vas deferens), deferens), Caylor Caylor cardiofacial cardiofacial syndrome (CBAVD), syndrome (CBAVD), CEPCEP (congenital (congenital

erythropoietic porphyria), cystic erythropoietic porphyria), cysticfibrosis, fibrosis,congenital congenital hypothyroidism, hypothyroidism, Chondrodystrophy Chondrodystrophy

syndrome (achondroplasia), otospondylomegaepiphyseal syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, dysplasia, Lesch-Nyhan syndrome, Lesch-Nyhan syndrome,

galactosemia, galactosemia, Ehlers-Danlos Ehlers-Danlos syndrome, Thanatophoric dysplasia, syndrome, Thanatophoric dysplasia, Coffin-Lowry Coffin-Lowry syndrome, syndrome, Cockayne syndrome, Cockayne syndrome, (familial (familial adenomatous adenomatous polyposis), polyposis), Congenital Congenital erythropoietic erythropoietic porphyria, porphyria,

Congenital heart disease, Congenital heart disease, Methemoglobinemia/Congenital Methemoglobinemia/Congenital methaemoglobinaemia, methaemoglobinaemia, achondroplasia, X-linkedsideroblastic achondroplasia, X-linked sideroblastic anemia, anemia,Connective Connective tissuedisease, tissue disease,Conotruncal Conotruncal anomaly anomaly

face face syndrome, Cooley'sAnemia syndrome, Cooley's Anemia (beta-thalassemia), (beta-thalassemia), Copper Copper storage storage disease disease (Wilson's (Wilson's disease), disease),

Copper transport disease Copper transport disease (Menkes disease), hereditary (Menkes disease), hereditarycoproporphyria, coproporphyria,Cowden syndrome, Cowden syndrome,

392

Craniofacial dysarthrosis (Crouzon (Crouzon syndrome), Creutzfeldt-Jakob disease disease (prion disease), 2023248067 10 2023

Craniofacial dysarthrosis syndrome), Creutzfeldt-Jakob (prion disease),

Cockayne syndrome,Cowden Cockayne syndrome, Cowden syndrome, syndrome, Curschmann-Batten-Steinert Curschmann-Batten-Steinert syndrome syndrome (myotonic (myotonic

Oct dystrophy), Beare-Stevenson cutis dystrophy), Beare-Stevenson cutis gyrata gyratasyndrome, syndrome, primary primary hyperoxaluria, hyperoxaluria, spondyloepimetaphysealdysplasia spondyloepimetaphyseal dysplasia (Strudwick (Strudwick type), type), muscular muscular dystrophy, dystrophy, Duchenne Duchenne and Becker and Becker

types (DBMD), types (DBMD), Usher Usher syndrome, syndrome, Degenerative Degenerative nerve nerve diseases diseases including including de Grouchy de Grouchy syndrome syndrome

and Dejerine-Sottas syndrome, and Dejerine-Sottas syndrome,developmental developmental disabilities,distal disabilities, distal spinal spinal muscular atrophy, type muscular atrophy, type V, V, androgen insensitivity syndrome, androgen insensitivity DiffuseGloboid syndrome, Diffuse GloboidBody Body Sclerosis Sclerosis (Krabbe (Krabbe disease), disease), Di George's Di George's

syndrome, Dihydrotestosterone syndrome, Dihydrotestosterone receptor receptor deficiency, deficiency, androgen androgen insensitivity insensitivity syndrome, syndrome, Down Down syndrome, Dwarfism, syndrome, Dwarfism, erythropoietic erythropoietic protoporphyria protoporphyria Erythroid Erythroid 5-aminolevulinate 5-aminolevulinate synthetase synthetase

deficiency, deficiency, Erythropoietic Erythropoietic porphyria, erythropoietic erythropoietic protoporphyria, protoporphyria, erythropoietic erythropoietic uroporphyria, uroporphyria,

Friedreich's ataxia,, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure Friedreich's ataxia,, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure

sensitive sensitive neuropathy, neuropathy, primary primary pulmonary hypertension (PPH), pulmonary hypertension (PPH), Fibrocystic Fibrocystic disease disease of of the the pancreas, fragile pancreas, fragile XXsyndrome, syndrome, galactosemia, galactosemia, genetic genetic brain disorders, brain disorders, Giant Giant cell cell hepatitis hepatitis

(Neonatal hemochromatosis), Gronblad-Strandberg (Neonatal hemochromatosis), Gronblad-Strandberg syndrome syndrome(pseudoxanthoma (pseudoxanthoma elasticum), elasticum),

Gunther disease(congenital Gunther disease (congenitalerythropoietic erythropoieticporphyria), porphyria),haemochromatosis, haemochromatosis, Hallgren Hallgren syndrome, syndrome,

sickle cell anemia, sickle cell hemophilia,hepatoerythropoietic anemia, hemophilia, hepatoerythropoieticporphyria porphyria (HEP), (HEP), Hippel-Lindau Hippel-Lindau disease disease

(von Hippel-Lindau (von Hippel-Lindau disease), disease), Huntington's Huntington's disease, disease, Hutchinson-Gilford Hutchinson-Gilford progeriaprogeria syndrome syndrome

(progeria), (progeria),Hyperandrogenism, Hyperandrogenism, Hypochondroplasia, Hypochromicanemia, Hypochondroplasia, Hypochromic anemia,Immune Immune system system

disorders, including disorders, including X-linked X-linkedsevere severe combined combined immunodeficiency, immunodeficiency, Insley-Astley Insley-Astley syndrome,syndrome,

Kennedy's syndrome, Kennedy's syndrome, Jackson-Weiss Jackson-Weisssyndrome, syndrome,Joubert Joubertsyndrome, syndrome,Lesch-Nyhan Lesch-Nyhan syndrome, syndrome,

Jackson-Weisssyndrome, Jackson-Weiss syndrome, Kidney Kidney diseases, diseases, including including hyperoxaluria, hyperoxaluria, Klinefelter's Klinefelter's syndrome, syndrome,

Kniest dysplasia, Kniest dysplasia, Lacunar Lacunar dementia,Langer-Saldino dementia,Langer-Saldino achondrogenesis, achondrogenesis, ataxia telangiectasia, ataxia telangiectasia,

Lynch syndrome, Lynch syndrome, Lysyl-hydroxylase Lysyl-hydroxylase deficiency, deficiency, Machado-Joseph Machado-Joseph disease, disease, Metabolic Metabolic disorders, disorders,

including Kniest dysplasia, including Kniest dysplasia, Marfan Marfansyndrome, syndrome, Movement Movement disorders, disorders, Mowat-Wilson Mowat-Wilson syndrome,syndrome,

cystic cystic fibrosis, fibrosis,Muenke syndrome,Multiple Muenke syndrome, Multipleneurofibromatosis, neurofibromatosis, Nance-Insley Nance-Insley syndrome, syndrome, Nance Nance-

Sweeney chondrodysplasia, Sweeney chondrodysplasia, Niemann-Pick Niemann-Pickdisease, disease,Noack Noack syndrome syndrome (Pfeiffer (Pfeiffer syndrome), syndrome),

Osler-Weber-Rendu disease, Osler-Weber-Rendu disease, Peutz-Jeghers Peutz-Jeghers syndrome, syndrome, Polycystic Polycystic kidney kidney disease, disease, polyostotic polyostotic

fibrous fibrous dysplasia (McCune-Albright dysplasia (McCune-Albright syndrome), syndrome), Peutz-Jeghers Peutz-Jeghers syndrome, syndrome, Prader-Labhart-Willi Prader-Labhart-Willi

syndrome, hemochromatosis, primary syndrome, hemochromatosis, primaryhyperuricemia hyperuricemiasyndrome syndrome (Lesch-Nyhan (Lesch-Nyhan syndrome), syndrome),

primary pulmonary primary pulmonary hypertension, hypertension, primary primary senile senile degenerative degenerative dementia, dementia, prionprion disease, disease, progeria progeria

(Hutchinson GilfordProgeria (Hutchinson Gilford ProgeriaSyndrome), Syndrome), progressive progressive chorea, chorea, chronic chronic hereditary hereditary (Huntington) (Huntington)

393

(Huntington's disease), progressive progressivemuscular muscular atrophy, spinalspinal muscular atrophy,atrophy, propionicpropionic 2023248067 10 Oct 2023

(Huntington's disease), atrophy, muscular

acidemia, protoporphyria, acidemia, protoporphyria, proximal proximalmyotonic myotonic dystrophy, dystrophy, pulmonary pulmonary arterial arterial hypertension, hypertension, PXE PXE (pseudoxanthoma elasticum), (pseudoxanthoma elasticum), Rb Rb (retinoblastoma), (retinoblastoma), Recklinghausen Recklinghausen disease disease (neurofibromatosis (neurofibromatosis

type I), type I), Recurrent Recurrent polyserositis, polyserositis,Retinal Retinaldisorders, disorders,Retinoblastoma, Retinoblastoma, Rett Rett syndrome, RFALS syndrome, RFALS type type

3, 3, Ricker syndrome,Riley-Day Ricker syndrome, Riley-Day syndrome, syndrome, Roussy-Levy Roussy-Levy syndrome, syndrome, severe achondroplasia severe achondroplasia with with developmental delay developmental delay and and acanthosis acanthosis nigricans nigricans(SADDAN), Li-Fraumeni syndrome, (SADDAN), Li-Fraumeni syndrome,sarcoma, sarcoma, breast, leukemia, breast, and adrenal leukemia, and adrenal gland gland(SBLA) (SBLA) syndrome, syndrome, sclerosis sclerosis tuberose tuberose (tuberous (tuberous sclerosis), sclerosis),

SDAT, SED SDAT, SED congenital(spondyloepiphyseal congenital (spondyloepiphyseal dysplasia dysplasia congenita), congenita), SED SEDStrudwick Strudwick (spondyloepimetaphyseal dysplasia, Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), type), SEDc (spondyloepiphyseal dysplasia SEDc (spondyloepiphyseal dysplasia congenita) SEMD, congenita) SEMD,Strudwick Strudwick typetype (spondyloepimetaphyseal (spondyloepimetaphyseal dysplasia, dysplasia, Strudwick Strudwick type), type),

Shprintzen syndrome, Shprintzen syndrome, Skin Skin pigmentation pigmentationdisorders, disorders, Smith-Lemli-Opitz Smith-Lemli-Opitz syndrome, syndrome,South- South African genetic African genetic porphyria porphyria(variegate (variegateporphyria), porphyria), infantile-onset infantile-onset ascending ascending hereditary hereditary spastic spastic

paralysis, Speech paralysis, andcommunication Speech and communication disorders, disorders, sphingolipidosis,Tay-Sachs sphingolipidosis, Tay-Sachs disease, disease,

spinocerebellar spinocerebellar ataxia, Stickler syndrome, ataxia, Stickler syndrome, stroke, stroke, androgen androgen insensitivitysyndrome, insensitivity syndrome, tetrahydrobiopterin deficiency, tetrahydrobiopterin deficiency, beta-thalassemia, beta-thalassemia,Thyroid Thyroid disease, disease, Tomaculous Tomaculous neuropathy neuropathy

(hereditary (hereditary neuropathy withliability neuropathy with liability to to pressure pressure palsies), palsies), Treacher Treacher Collins Collins syndrome, TriploX X syndrome, Triplo

syndrome syndrome ( (triple triple X syndrome),Trisomy X syndrome), Trisomy21 21 (Down (Down syndrome), syndrome), Trisomy Trisomy X, VHL X, VHL syndrome syndrome (von (von Hippel-Lindaudisease), Hippel-Lindau disease),Vision Visionimpairment impairment and and blindness blindness (Alstr6m (Alström syndrome), syndrome), VrolikVrolik disease, disease,

Waardenburg syndrome, Waardenburg syndrome, Warburg Warburg Sjo Fledelius Sjo Fledelius Syndrome, Syndrome, Weissenbacher-Zweymiiller Weissenbacher-Zweymiller

syndrome, Wolf-Hirschhorn syndrome, syndrome, Wolf-Hirschhorn syndrome,Wolff Wolff Periodicdisease, Periodic disease, Weissenbacher-Zweymüiller Weissenbacher-Zweymiiller syndrome and Xeroderma syndrome and Xeroderma pigmentosum. pigmentosum.

[0915] any aspect In aspect

[0915] In any or embodiment or embodiment described herein,herein, described the composition further further the composition comprises comprises an an additionalbioactive additional bioactive agent. agent.

[0916] any aspect In aspect

[0916] In any or embodiment or embodiment described described herein, herein, the the additional additional bioactive agent is agent bioactive at is at least least one of an one of an anti-cancer anti-cancer agent, agent, an ananti-neurodegenerative anti-neurodegenerativeagent, agent,an an antimicrobial antimicrobial agent, agent, an an

antiviral agent, antiviral agent, anananti-HIV anti-HIV agent, agent, an antifungal an antifungal agent,agent, or a combination or a combination thereof. thereof.

[0917] any aspect In aspect

[0917] In any or embodiment or embodiment described herein, herein, described the anticancer the anticancer agent agent is is selected selected from from the group the group consisting consisting of of everolimus, everolimus, trabectedin, trabectedin, abraxane, abraxane, TLK 286, AV-299, TLK 286, AV-299,DN-101, DN-101, pazopanib, GSK690693, pazopanib, RTA744, GSK690693, RTA 744,ONON 0910.Na,AZD 0910.Na, AZD 62446244 (ARRY-142886), (ARRY-142886), AMN-107, AMN-107, TKI- TKI 258, GSK461364, 258, AZD GSK461364, AZD 1152, 1152, enzastaurin, vandetanib, enzastaurin, vandetanib, ARQ-197, MK-0457,MLN8054, ARQ-197, MK-0457, MLN8054, PHA-PHA

739358, R-763, AT-9263, 739358, R-763, FLT-3inhibitor, AT-9263, aa FLT-3 inhibitor, aa VEGFR inhibitor, an VEGFR inhibitor, an EGFR EGFR TKTK inhibitor, an inhibitor, an

394 aurora kinase kinase inhibitor, inhibitor, aa PIK-1 PIK-1modulator, modulator, a Bcl-2 inhibitor, an HDAC inhbitor, a c-METa c-MET 2023248067 10 Oct 2023 a Bcl-2 inhibitor, an HDAC inhbitor, inhibitor, inhibitor, aa PARP inhibitor, aa Cdk PARP inhibitor, Cdkinhibitor, inhibitor, an an EGFR EGFR TK TK inhibitor, inhibitor, an an IGFR-TK IGFR-TK inhibitor, inhibitor, an an anti-HGF antibody, aa PI3 anti-HGF antibody, P13 kinase kinase inhibitors, inhibitors, ananAKT inhibitor, an AKT inhibitor, an mTORC1/2 inhibitor, aa mTORC1/2 inhibitor,

JAK/STAT JAK/STAT inhibitor, inhibitor, a checkpoint-i a checkpoint-1 or 2 or 2 inhibitor, inhibitor, a focal a focal adhesion adhesion kinase kinase inhibitor, inhibitor, a Mapa Map kinase kinase kinase kinase (mek) (mek) inhibitor, inhibitor, a VEGF trapantibody, VEGF trap antibody,pemetrexed, pemetrexed, erlotinib,dasatanib, erlotinib, dasatanib,nilotinib, nilotinib, decatanib, panitumumab, decatanib, panitumumab, amrubicin, amrubicin, oregovomab, oregovomab, Lep-etu,Lep-etu, nolatrexed, nolatrexed, azd2171, azd2171, batabulin, batabulin,

ofatumumab, zanolimumab, ofatumumab, zanolimumab, edotecarin,tetrandrine, edotecarin, tetrandrine,rubitecan, rubitecan,tesmilifene, tesmilifene, oblimersen, oblimersen, ticilimumab, ipilimumab, ticilimumab, ipilimumab, gossypol, gossypol,Bio Bio111, 131-I-TM-601 ,ALT-110, 111,131-I-TM-601 ALT-110,BIO BIO 140, 140, CC CC 8490, 8490,

cilengitide, cilengitide, gimatecan, IL13-PE38QQR, gimatecan, IL13-PE38QQR,INO 1001,, IPdR INO 1001 KRX-0402,lucanthone, IPdR 1KRX-0402, lucanthone, LY LY317615, 317615, neuradiab, vitespan, neuradiab, vitespan,Rta Rta744, 744,Sdx Sdx 102, 102, talampanel, talampanel,atrasentan, 311311, romidepsin, Xr Xr atrasentan, romidepsin, ADS ADS-

100380, sunitinib, 5-fluorouracil, 100380, sunitinib, 5-fluorouracil, vorinostat, vorinostat, etoposide, etoposide, gemcitabine, gemcitabine,doxorubicin, doxorubicin, liposomal liposomal

doxorubicin, 5'-deoxy-5-fluorouridine, doxorubicin, 5'-deoxy-5-fluorouridine,vincristine, vincristine, temozolomide, temozolomide, ZK-304709, ZK-304709, seliciclib;seliciclib;

PD0325901, , AZD-6244, PD0325901AZD-6244, capecitabine, capecitabine, L-Glutamic L-Glutamic acid, Nacid, N -[4-[2-(2-amino-4,7-dihydro-4-oxo-i -[4-[2-(2-amino-4,7-dihydro-4-oxo-1

H -- pyrrolo[2,3- H pyrrolo[2,3- dd ]pyrimidin-5-yl)ethyl]benzoyl], ]pyrimidin-5-yl)ethyl]benzoyl]-,disodium disodium salt, salt, heptahydrate, heptahydrate, camptothecin, camptothecin,

PEG-labeledirinotecan, PEG-labeled irinotecan,tamoxifen, tamoxifen, toremifene toremifene citrate, citrate, anastrazole, anastrazole, exemestane, exemestane, letrozole, letrozole,

DES(diethylstilbestrol), estradiol, estrogen, DES(diethylstilbestrol), estradiol, conjugated estrogen, estrogen, conjugated estrogen,bevacizumab, bevacizumab, IMC-iCii IMC-1C11 ,

, CHIR-258,); 3-[5-(methylsulfonylpiperadinemethyl)- CHIR-258,); 3-[5-(methylsulfonylpiperadinemethyl)- indolylj-quinolone, indolylj-quinolone, vatalanib, vatalanib, AG-013736, AG-013736,

AVE-0005,thethe AVE-0005, acetatesalt acetate saltof of[D-

[D- Ser(Bu Ser(Bu6 t,Azgly ) 6,Azgly 10 ] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu 10 (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu

t )-Leu-Arg-Pro- )-Leu-Arg-Pro- Azgly-NH acetate[C Azgly-NH 22 acetate 9H 4Ni-(CHO)x

[C9HNOi 8 Oi4 -(C2H X where 4 02)x where = I togoserelin = 1 tox2.4], 2.4], goserelin acetate, leuprolide acetate, leuprolide acetate, acetate,triptorelin triptorelinpamoate, pamoate, medroxyprogesterone medroxyprogesterone acetate, acetate, hydroxyprogesterone caproate, hydroxyprogesterone caproate, megestrol megestrol acetate, acetate, raloxifene, raloxifene, bicalutamide, bicalutamide, flutamide, flutamide, nilutamide, megestrol nilutamide, megestrol acetate, acetate, CP-724714; CP-724714;TAK-165, TAK-165, HKI-272, HKI-272, erlotinib, erlotinib, lapatanib, lapatanib, canertinib, canertinib,

ABX-EGF ABX-EGF antibody, antibody, erbitux,EKB-569, erbitux, EKB-569, PKI-166, PKI-166, GW-572016, GW-572016, Ionafarnib, Ionafarnib, BMS-214662, BMS-214662,

tipifarnib; tipifarnib;amifostine, amifostine,NVP-LAQ824, suberoylanalide NVP-LAQ824, suberoyl analide hydroxamic hydroxamic acid, acid, valproic valproic acid, acid,

trichostatin A,A, FK-228, trichostatin SUi1248,sorafenib, FK-228, SU11248, sorafenib, KRN951 KRN951 , aminoglutethimide, aminoglutethimide, arnsacrine, arnsacrine,

anagrelide, L-asparaginase, Bacillus anagrelide, L-asparaginase, Bacillus Calmette-Guerin Calmette-Guerin (BCG) (BCG) vaccine, vaccine, adriamycin, adriamycin, bleomycin, bleomycin,

buserelin, busulfan, buserelin, busulfan, carboplatin, carboplatin, carmustine, carmustine,chlorambucil, chlorambucil, cisplatin, cisplatin, cladribine,clodronate, cladribine, clodronate, cyproterone, cytarabine, cytarabine, dacarbazine, dacarbazine, dactinomycin, dactinomycin,daunorubicin, daunorubicin,diethylstilbestrol, diethylstilbestrol, epirubicin, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, fludarabine, fludrocortisone, fluoxymesterone,flutamide, flutamide,gleevac, gleevac,gemcitabine, gemcitabine, hydroxyurea, hydroxyurea,

idarubicin, idarubicin, ifosfamide, ifosfamide, imatinib, imatinib,leuprolide, leuprolide,levamisole, levamisole,lomustine, lomustine,mechlorethamine, melphalan, mechlorethamine, melphalan,

6-mercaptopurine, mesna, 6-mercaptopurine, mesna, methotrexate, methotrexate, mitomycin, mitomycin, mitotane, mitotane,mitoxantrone, mitoxantrone,nilutamide, nilutamide,

395 octreotide, oxaliplatin,pamidronate, pamidronate, pentostatin, 2023248067 10 Oct 2023 octreotide, oxaliplatin, pentostatin, plicamycin, plicamycin, porfimer, porfimer, procarbazine, procarbazine, raltitrexed,raltitrexed, rituximab, streptozocin, rituximab, streptozocin, teniposide, teniposide, testosterone, testosterone, thalidomide, thalidomide, thioguanine, thioguanine, thiotepa, thiotepa, tretinoin, tretinoin, vindesine, 13-cis-retinoic vindesine, 13-cis-retinoic acid, acid, phenylalanine phenylalanine mustard, mustard, uracil mustard, uracil mustard, estramustine, estramustine, altretamine,altretamine, floxuridine, floxuridine, 5-deooxyuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, deoxycoformycin, calcitriol, calcitriol,valrubicin, valrubicin, mithramycin, vinblastine, vinorelbine, mithramycin, vinblastine, vinorelbine, topotecan, topotecan,razoxin, razoxin,marimastat, marimastat, COL-3, neovastat, BMS-275291 COL-3, neovastat, , squalamine,endostatin, BMS-275291, squalamine, endostatin, SU5416, SU5416,SU6668, SU6668, EMD121974, EMD121974, interleukin-12, IM862, interleukin-12, IM862, angiostatin, angiostatin, vitaxin, vitaxin, droloxifene, droloxifene, idoxyfene, idoxyfene, spironolactone, spironolactone, finasteride, finasteride, cimitidine, trastuzumab, cimitidine, trastuzumab, denileukin denileukindiftitox,gefitinib, diftitox,gefitinib, bortezimib, bortezimib,paclitaxel, paclitaxel, cremophor-free cremophor-free paclitaxel, docetaxel, paclitaxel, epithilone docetaxel, B, B, epithilone BMS- 247550, BMS-310705, BMS- 247550, BMS-310705, droloxifene, droloxifene, 4-4 hydroxytamoxifen,pipendoxifene, hydroxytamoxifen, pipendoxifene, ERA- ERA- 923, 923, arzoxifene, arzoxifene, fulvestrant, fulvestrant, acolbifene, acolbifene, lasofoxifene, lasofoxifene, idoxifene, idoxifene,TSE-424, TSE-424, HMR- 3339,ZK186619, HMR- 3339, ZK186619, topotecan,PTK787/ZK topotecan, PTK787/ZK 222584, 222584, VX-745, VX-745, PD PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin 184352, rapamycin, 40-0-(2-hydroxyethyl)-rapamycin, temsirolimus, temsirolimus, AP-23573, AP-23573, RAD001 RADOOl

, ABT-578, BC-210,LY294002, ABT-578, BC-210, LY294002, LY292223, LY292223, LY292696, LY292696, LY293684, LY293684, LY293646, LY293646, wortmannin, wortmannin,

ZM336372,L-779,450, ZM336372, L-779,450,PEG-filgrastim, PEG-filgrastim,darbepoetin, darbepoetin,erythropoietin, erythropoietin, granulocyte granulocyte colony- colony stimulating factor, factor, zolendronate, zolendronate,prednisone, prednisone, cetuximab, cetuximab, granulocyte granulocyte macrophage macrophage colony- colony stimulating factor,histrelin, stimulating factor, histrelin,pegylated pegylated interferon interferon alfa-2a, alfa-2a, interferon interferon alfa-2a, alfa-2a, pegylated pegylated interferon interferon

alfa-2b, interferon alfa-2b, interferon alfa-2b, alfa-2b,azacitidine, azacitidine,PEG-L-asparaginase, PEG-L-asparaginase, lenalidomide, lenalidomide, gemtuzumab, gemtuzumab,

hydrocortisone, interleukin-11, hydrocortisone, interleukin-11 , dexrazoxane, dexrazoxane,alemtuzumab, alemtuzumab, all-transretinoicacid, all-transretinoic acid,ketoconazole, ketoconazole, interleukin-2, interleukin-2, megestrol, immuneglobulin, megestrol, immune globulin, nitrogen nitrogen mustard, mustard, methylprednisolone, methylprednisolone,

ibritgumomab tiuxetan,androgens, ibritgumomab tiuxetan, androgens,decitabine, decitabine,hexamethylmelamine, hexamethylmelamine, bexarotene, bexarotene, tositumomab, tositumomab,

arsenic arsenic trioxide, trioxide, cortisone, cortisone,editronate, editronate,mitotane, mitotane,cyclosporine, cyclosporine,liposomal liposomal daunorubicin, daunorubicin, Edwina Edwina-

asparaginase, strontium 89, asparaginase, strontium 89, casopitant, casopitant, netupitant, netupitant, an an NK-1 NK-1receptor receptorantagonists, antagonists,palonosetron, palonosetron, aprepitant, aprepitant, diphenhydramine, hydroxyzine,metoclopramide, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, lorazepam, alprazolam, alprazolam, haloperidol, haloperidol,

droperidol, dronabinol, dexamethasone, droperidol, dronabinol, dexamethasone, methylprednisolone, methylprednisolone, prochlorperazine, prochlorperazine, granisetron, granisetron,

ondansetron, dolasetron, ondansetron, dolasetron, tropisetron, tropisetron, pegfilgrastim, pegfilgrastim, erythropoietin, erythropoietin, epoetin epoetin alfa, alfa, darbepoetin darbepoetin alfa alfa and mixtures thereof. and mixtures thereof.

[0918] An An

[0918] additional additional aspect aspect of of thethe presentdisclosure present disclosureprovides providesa amethod method forfor inducing inducing

degradation ofa atarget degradation of target protein proteininina acell cellcomprising comprising administering administering an effective an effective amount amount of a of a

compound compound of of thepresent the presentdisclosure disclosuretotothe the cell, cell, wherein the compound wherein the effectuatesdegradation compound effectuates degradationof of the target the target protein. protein.

396

[0919] Another

[0919] Another aspect aspect of the of the present present disclosureprovides disclosure providesa acomposition compositioncomprising comprisingan an effective effective amount of aa compound amount of compound of of thepresent the presentdisclosure disclosurefor foruse use in in aa method fortreating method for treating cancer, cancer, said method comprisingadministering method comprising administering thethe composition composition to atopatient a patient in in need need thereof, thereof, wherein wherein the the

compositionisis effectuates composition effectuates for for the the treatment treatment or or alleviation alleviation of of at atleast leastone onesymptom ofcancer symptom of cancerinin the patient. the patient.

[0920] In any

[0920] In any aspect aspect or embodiment or embodiment described described herein, herein, the cancer the cancer is squamous-cell is squamous-cell carcinoma, carcinoma,

basal cell basal cell carcinoma, carcinoma, adenocarcinoma, adenocarcinoma, hepatocellular hepatocellular carcinomas, carcinomas, and renal and renal cell carcinomas, cell carcinomas,

cancer of of the the bladder, bladder, bowel, bowel, breast, breast, cervix, cervix, colon, colon, esophagus, esophagus, head, head,kidney, kidney,liver, liver, lung, lung, neck, neck, ovary, pancreas, prostate, ovary, pancreas, prostate, and stomach; leukemias; and stomach; leukemias; benign benignandand malignant malignant lymphomas, lymphomas,

particularly Burkitt's particularly Burkitt'slymphoma and Non-Hodgkin's lymphoma and Non-Hodgkin'slymphoma; lymphoma; benign benign and malignant and malignant

melanomas; myeloproliferative melanomas; myeloproliferative diseases; diseases; multiple multiple myeloma, sarcomas, including myeloma, sarcomas, including Ewing's Ewing's sarcoma, hemangiosarcoma,Kaposi's sarcoma, hemangiosarcoma, Kaposi's sarcoma, sarcoma, liposarcoma, liposarcoma, myosarcomas, myosarcomas, peripheral peripheral

neuroepithelioma, synovialsarcoma, neuroepithelioma, synovial sarcoma,gliomas, gliomas,astrocytomas, astrocytomas, oligodendrogliomas, oligodendrogliomas, ependymomas, ependymomas,

gliobastomas, neuroblastomas,ganglioneuromas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, gangliogliomas, medulloblastomas, medulloblastomas, pinealpineal cell cell

tumors, meningiomas, tumors, meningeal sarcomas, meningiomas, meningeal sarcomas, neurofibromas, neurofibromas, and and Schwannomas; bowelcancer, Schwannomas; bowel cancer, breast cancer, breast cancer, prostate prostate cancer, cancer, cervical cervical cancer, cancer,uterine uterinecancer, cancer,lung lung cancer, cancer, ovarian ovarian cancer, cancer,

testicular cancer, testicular cancer, thyroid thyroid cancer, astrocytoma, esophageal cancer, astrocytoma, esophagealcancer, cancer,pancreatic pancreaticcancer, cancer, stomach stomach

cancer, cancer, liver liver cancer, cancer,colon colon cancer, cancer,melanoma; carcinosarcoma,Hodgkin's melanoma; carcinosarcoma, Hodgkin's disease, disease, Wilms' Wilms' tumor tumor

or teratocarcinomas, T-lineage or teratocarcinomas, T-lineage Acute Acute lymphoblastic lymphoblastic Leukemia Leukemia (T-ALL), (T-ALL), T-lineageT-lineage

lymphoblastic Lymphoma lymphoblastic Lymphoma (T-LL), (T-LL), Peripheral Peripheral T-cellT-cell lymphoma, lymphoma, Adult Leukemia, Adult T-cell T-cell Leukemia, Pre-B Pre-B ALL, Pre-B ALL, Pre-B Lymphomas, Lymphomas,Large LargeB-cell B-cell Lymphoma, Lymphoma, BurkittsLymphoma, Burkitts Lymphoma, B-cellALL, B-cell ALL, Philadelphia chromosome Philadelphia chromosome positive positive ALLALL and and Philadelphia Philadelphia chromosome chromosome positive positive CML. CML.

[0921] WhileWhile

[0921] preferred preferred embodiments embodiments of the of the invention invention haveshown have been beenand shown and described described herein, herein, it it will willbe beunderstood understood that that such such embodiments embodiments areare provided provided by by wayway of example of example only. only. Numerous Numerous

variations, changes variations, changes andand substitutions substitutions will will occuroccur to skilled to those those skilled in the in the art art without without departingdeparting from from the spirit the spiritof ofthe theinvention. invention. Accordingly, it is Accordingly, it is intended intended that that the the appended claims cover appended claims coverall all such such variations asfall variations as fall within withinthe thespirit spiritand and scope scope of the of the invention. invention.

[0922]

[0922] EXAMPLES EXAMPLES

[0923] A. A.

[0923] Protein Protein Degradation Degradation Bioassays: Bioassays:

397

Oct 2023 The following

[0924] The following

[0924] bioassays bioassays evaluate evaluate the level the level of protein of protein degradation degradation observed observed in various in various

cell cell types typesusing using representative representativecompounds disclosedherein. compounds disclosed herein.

[0925] InIneach

[0925] bioassay, cells eachbioassay, treated with were treated cells were varying amounts with varying of compounds amounts of compounds encompassedby by encompassed the the present present disclosure. disclosure. The degradation The degradation of the of the following following proteinsproteins may be may be 2023248067 10

evaluated: estrogen evaluated: estrogenreceptor a receptor (ERa), bromodomain-containing protein (ER), bromodomain-containing protein 44 (BRD4), (BRD4), androgen androgen receptor (AR), receptor (AR), and and BRaf BRafprotein. protein.

1. ERE

[0926] 1. ERE

[0926] Luciferase Luciferase Assay for for Assay compounds compounds in Table in Table 5. 5.

[0927] T47D-KBluc

[0927] T47D-KBluc cellscells (ATCC© (ATCC® #CRL_2865, #CRL_2865, T47D human T47D breast cancercancer human breast cellscells stably stably transfected with transfected with estrogen estrogen responsive element/promoter/luciferase reporter responsive element/promoter/luciferase reporter gene) gene) were wereseeded seededinto into 96-well white opaque 96-well white opaqueplates platesin in RPMI RPMI growth growth medium medium supplemented supplemented with with 10% 10%bovine fetal fetal bovine serum (FBS)and serum (FBS) andallowed allowed to to adhere adhere overnight overnight in in a 37°C a 37°C humidified humidified incubator. incubator. TheThe following following day,day,

cells cells were were treated treated with with PROTACs PROTACs in in a 12-point a 12-point concentration concentration curve curve (top (top finalconcentration final concentrationofof 300 nMwith 300 nM withsubsequent subsequent concentrations concentrations being being 3-fold 3-fold lesswith less with2 2pMpM being being thethe lowest lowest

concentration in concentration in the the assay). assay). Each PROTAC Each PROTAC was was tested tested independently independently in experiments in two two experiments on 96-on 96 well plates. well plates. After After24 24hours, hours,media media was was removed andlysis removed and lysisbuffer buffer was wasadded addedtotothe thewells. wells. Followinglysis, Following lysis, Bright-GloTM Luciferase Bright-Glo Luciferase Assay Assay Substrate Substrate (Promega, (Promega, Madison Madison WI) WI) was was added added

and the and the luciferase luciferase activity activitywas wasmeasured using aa Cytation measured using 3 plate Cytation 3 plate reader reader (BioTekTM, Winooski, (BioTekM, Winooski,

VT). Each VT). Eachcompound compound was was assayed assayed in duplicate in duplicate and activity and the the activity was was calculated calculated as IC50 as IC50 using using

GraphPad Prism GraphPad Prism software software (San (San Diego, Diego, CA). CA).

[0928] 2. 2.

[0928] Estrogen Estrogen Receptor-alpha Receptor-alpha (ERa) (ER) degradation degradation assayassay in MCF-7 in MCF-7 cellscells usingusing westernblot western blotmethod methodforfor Table Table 5. 5.

[0929] The exemplary

[0929] The exemplary novel novel ERa degraders ER degraders were for were assessed assessed their for their activity activity in degrading in degrading

ERa ER in in MCF-7 MCF-7 cells cells viavia western western blot. blot. TheThe assay assay waswas carried carried outout in in thepresence the presence of of 10% 10% FBS FBS or or high percentage high percentage of of human humanorormouse mouse serum. serum. Protocols Protocols of the of the western western blot blot assay assay areare described described

below. below.

[0930] MCF7

[0930] MCF7 cellscells werewere grown grown in DMEM/F12 in DMEM/F12 with FBS with 10% 10%and FBS and seeded seeded at 24,000 at 24,000 cells cells perper

well inin100 well 100µlpl into into 96-well 96-well clearclear tissue tissue culture culture plates. plates. The following The following day, were day, the cells the treated cells were treated with PROTACs with PROTACs in a in a 7-point 7-point concentration concentration curvecurve with with 100 nM100 nMthe being being top the top concentration concentration and and serial serial dilutions dilutionstotomake make the the other other concentrations concentrations (30 nM, 10nM, nM, 10 nM,3 nM, 3 nM, 1 nM, 1 nM, and nM). and 0.3 0.3 nM). At At all concentrations, all concentrations, 0.01% DMSO 0.01% DMSO is the is the final final concentration concentration in in thethe well. well. The The following following day, day, the the plates are plates are aspirated, aspirated,washed washed with 50 50 pl of cold PBS. µl of Thecells PBS. The cells are are lysed with 50 50pl/well µl/well 4°C Cell 4°C Cell

398

Lysis Buffer (Catalog#9803; Buffer (Catalog# 9803;Cell CellSignaling SignalingTechnology, Technology, Danvers, MA) MA) (20mM (20mM Tris-HCLTris-HCL (pH 2023248067 10 Oct 2023

Lysis Danvers, (pH

7.5),150 7.5), 150mM mM NaCl, 1mM Na2EDTA, 1mM NaEDTA, 1 mM1 EGTA, mM EGTA, 1% Triton, 1% Triton, 2.5 mM2.5sodium mM sodium pyrophosphate, pyrophosphate,

1 mM B-glycerophosphate, mM B-glycerophosphate, 1 mM1 sodium mM sodium vanadate, vanadate, 1 ug/ml1 leupeptin). ug/ml leupeptin). Lysates Lysates were clarified were clarified

at 16,000 xx ggfor at 16,000 for1010minutes, minutes,andand 2 tg 2 µg of protein of protein was was subjected subjected to SDS-PAGE to SDS-PAGE analysis analysis and and followed byimmunoblotting followed by immunoblotting according according to standard to standard protocols. protocols. The antibodies The antibodies usedERwere ERU used were

(Cell Signaling Signaling Technologies TechnologiesCatalog Catalog #8644), #8644), and and Tubulin Tubulin (Sigma (Sigma Catalog Catalog #T9026; #T9026; St. Louis, St. Louis,

MO). Detection MO). Detection reagents reagents were Clarity Western were Clarity ECLsubstrate Western ECL substrate (Bio-Rad (Bio-Rad Catalog Catalog #170-5060; #170-5060; Hercules, CA). Hercules, CA).

[0931] MCF7 Alternatively, MCF7

[0931] Alternatively, cellswere cells grownininDMEM/F12 weregrown DMEMIF12 10% 10% withwith FBS and and seeded FBSseeded at at 24,000cells 24,000 cellsper perwell well in in 500500 pl24-well µl in in 24-well clear clear tissuetissue culture culture plates.plates. The following The following day, day, the cells the cells were treated were treated with with PROTACs PROTACs in ain5-point a 5-point concentration concentration curve curve (100 (100 nM, nM, 33 nM,3311nM, nM, 11 3.7nM, nM, 3.7 nM, and 1.2 nM) and 1.2 in the nM) in the presence presence of of 0.01% DMSO.After 0.01% DMSO. After 72 72 hours, hours, thethe wellsare wells areaspirated aspirated and and washed with washed with 500 500µlglofofPBS. PBS.TheThe cellsare cells arelysed lysedwith with100 100µl/well gl/well4°C 4°CCell CellLysis LysisBuffer Buffer (Catalog# 9803;Cell (Catalog# 9803; CellSignaling SignalingTechnology, Technology, Danvers, Danvers, MA)MA) (20mM(20mM Tris-HCL Tris-HCL (pH (pH 7.5), 1507.5), mM 150 mM

NaCl, 1mM NaCl, Na2EDTA, 1mM NaEDTA, 1 mM 1 mM EGTA, EGTA, 1% Triton, 1% Triton, 2.52.5 mMmM sodium sodium pyrophosphate, 1 1mM pyrophosphate, mMB-B glycerophosphate, 1 1mMmM glycerophosphate, sodium sodium vanadate, vanadate, 1 ug/ml 1 ug/ml leupeptin). leupeptin). Lysates Lysates were clarified were clarified at 16,000 at 16,000

x gg for X for 10 10 minutes, minutes, and and22µgpgofofprotein proteinwas was subjected subjected to to SDS-PAGE SDS-PAGE analysis analysis and followed and followed by by immunoblotting according immunoblotting according to to standard standard protocols.TheThe protocols. antibodies antibodies used used were were ERa Signaling ER (Cell (Cell Signaling TechnologiesCatalog Technologies Catalog#8644), #8644), andand Tubulin Tubulin (Sigma (Sigma Catalog Catalog #T9026; #T9026; St. Louis, St. Louis, MO). Detection MO). Detection

reagents were reagents Clarity Western were Clarity WesternECL ECL substrate(Bio-Rad substrate (Bio-Rad Catalog Catalog #170-5060; #170-5060; Hercules, Hercules, CA). CA).

[0932] 3. 3.

[0932] Estrogen Estrogen receptor-alpha receptor-alpha (ERa) (ER) degradation degradation assay assay using using In-Cell In-Cell WesternTM Western

Assay forTable Assay for Table5.5.

[0933] Degradation

[0933] Degradation of of ERERa by claimed by claimed compounds compounds werewere determined determined in MCF7 in MCF7 cells cells usinganan using

In-Cell Western In-Cell T Massay. Western assay. Briefly, Briefly, MCF7 MCF7 cellscells werewere plated plated in 96-well in 96-well plates plates (2000 (2000 cells cells perper well well

in 100 gl µl media) and incubated media) and incubatedatat 37°C 37°Cunder underananatmosphere atmosphereof of 5% 5% C02 CO in in a humidified a humidified

incubator overnight. incubator overnight. One-hundred One-hundred (100) (100) µl gl of of media media containing containing test test compound compound (at (at 2x 2x concentration) was concentration) was added added toappropriate to the the appropriate wells wells to to provide provide 11 decreasing 11 serially serially decreasing concentrations(top concentrations (top final final concentration, concentration, 1 gM3-fold 1 µM then then less 3-fold for less for the the next next 10 concentrations); 10 concentrations); a a vehicle control vehicle control (DMSO) was (DMSO) was also also added added forfor each each compound. compound. For each For each experiment, experiment, all all compounds compounds were were assayed assayed in duplicate in duplicate plates.Cells plates. Cellswere werethen thenincubated incubatedforfor3 3oror5 5days daysininthe the above-mentioned environment. above-mentioned environment. TheThe assay assay was was terminated terminated by removal by removal of media, of media, a single a single wash wash

399 with ice-cold ice-cold PBS andthe addition of theaddition of 50 50 µl pl paraformaldehyde paraformaldehyde(PFA: (PFA: 4% 4% AfterAfter in PBS). 15 2023248067 10 Oct 2023 with PBS and in PBS). 15 minutes in minutes in PFA PFAatatroom temperature,thethecells roomtemperature, cellswere werepermeabilized permeabilizedin inTris-phosphate-buffered Tris-phosphate-buffered saline saline with with Tween (0.1%)(TBST) Tween (0.1%) (TBST) supplemented supplemented with with Triton Triton X-100X-100 (0.5%)(0.5%) for 15for 15 minutes. minutes. Cells Cells were then were then blocked blockedinin BSA BSA(TBST (TBST withwith BSA,BSA, 3%)one 3%) for forhour. one hour. Primary Primary antibodies antibodies for thefor the detection of ERa (rabbitmonoclonal, ER (rabbit monoclonal,1:1000, 1:1000, Cell Cell Signaling Signaling Technology Technology Catalog Catalog #8644) #8644) and and tubulin (mouse tubulin monoclonal,1:5000, (mouse monoclonal, 1:5000,Sigma Sigma Catalog Catalog #T6074) #T6074) in TBST in TBST with with BSA BSA (3%) (3%) were were added. The added. Thecells cells were wereincubated incubatedovernight overnightatat4°C. 4°C.The Thecells cellswere werethen thenwashed washed thricewith thrice withTBST TBST at at room temperature and room temperature andthen thenincubated incubatedwith withanti-rabbit anti-rabbit and andanti-mouse anti-mousefluorescently-labelled fluorescently-labelled secondary antibodies (IRDye; secondary antibodies (IRDye©; LI-COR; LI-COR; Lincoln, Lincoln, NE) NE) in in LI-COR LI-COR blocking blocking buffer (Catalog buffer (Catalog

#927-50000)for #927-50000) forone onehour houratatroom room temperature.Following temperature. Following 3 washes 3 washes withwith TBST, TBST, the buffer the buffer was was removedand removed andthe theplates plateswere wereread readononananOdyssey® Odyssey© infrared infrared imaging imaging system (LI-COR); system Lincoln, (LI-COR©; Lincoln, NE) at NE) at 700 700 nm nmand and800 800 nm. nm. Using Using commercial commercial software software (ImageStudi (ImageStudioM; TM ; LI-COR, LI-COR, Lincoln, Lincoln, NE), NE), the staining the stainingintensity intensityfor forERERa and tubulin and tubulin inwell in each eachwaswell was quantified quantified andforexported and exported analysis.for analysis. For each For each data data point, point, ERa intensitywas ER intensity wasnormalized normalizedto to tubulinintensity tubulin intensityand andfor for each each compound compoundallall

normalized intensity normalized intensity values values were were normalized normalizedtotothe the vehicle vehicle control. control. DC o and DC 5and Dmax values D values were were determined followinga a4-parameter determined following 4-parameterICICcurve 5 o curve fit fit using using ACAS ACAS dose dose response response modulemodule (McNeil(McNeil

& CoInc.). & Co Inc.).

[0934] 4. 4.

[0934] AR AR ELISA AssayAssay ELISA Protocol Protocol for Table for Table 6 6

[0935] Compounds

[0935] Compounds were were evaluated evaluated in this in this assayassay in LNCaP in LNCaP and/orand/or VCaP utilizing VCaP cells cells utilizing similar protocols. The similar protocols. Theprotocols protocolsused used with with VCaPVCaP cells cells are described are described below. below. The androgen The androgen

receptor ELISA receptor assay was ELISA assay was performed performed using using PathScan PathScanARAR Sandwich Sandwich ELISA ELISA (Cell(Cell Signaling Signaling

Catalog#12850) according Catalog#12850) according to to thefollowing the followingassay assaysteps: steps:

[0936] cellscells VCaP

[0936] VCaP werewere seeded seeded at 40,000 at 40,000 cells/well at aa volume cells/well at 100 µL/well of 100 volume of in VCaP pL/well in VCaP assay medium assay medium [Phenol

[Phenol redred free free RPMI RPMI (Gibco (Gibco Cat#11835-030); Cat#11835-030); 5% Charcoal 5% Charcoal Stripped Stripped (Dextran (Dextran treated) FBS treated) (Omega FBS (Omega Scientific, Scientific, Cat#FB-04); Cat#FB-04); 1% penstrep 1% penstrep (Life Technologies,Gibco (Life Technologies, Gibco Cat#: Cat#: 10378-016)] in Corning 10378-016)] in Corning3904 3904 plates.TheThe plates. cells cells were were incubated incubated for for a minimum a minimum of 3 days. of 3 days. Cells Cells

were dosed were dosed with with PROTACs diluted inin 0.01% PROTACs diluted 0.01%DMSO DMSOandand thethe drug drug treatmentwas treatment wasallowed allowedfor for 55 hours. hours.

[0937] AR ELISA

[0937] AR ELISA (Cell Signaling) (Cell Signaling) was performed was performed as 1x as follows. follows. 1x Cell Cell Cell Signaling Signaling lysis Cell lysis buffer was buffer made (Catalogue was made (Catalogue #9803; #9803; comes comeswith withthe thekit). kit). Media Mediafrom from thethe treated wells treated wells isis aspirated, and aspirated, and100100 µL lxLcell lx cell lysis lysis buffer/well buffer/well is added. is added. Thewere The cells cells wereonplaced placed a shakeron a shaker for 10 for 10

400 minutes at at 4°C. 4°C. Twenty Twentymicroliters microlitersofoflysate lysate was wastransferred transferred to to 100µl 100u1of Diluentinin ELISA ELISA plate 2023248067 10 Oct 2023 minutes of Diluent plate

(0.15ug/ml -0.075 (0.15pg/ml 0.075µg/ml). pg/ml).The Thelysate-diluent lysate-diluent mixture mixture was was shaken shaken for for 30 30 minutes minutes at at 37°C. 37°C. Allow mouse Allow mouseARAR antibody,anti-mouse antibody, anti-mouse antibody, antibody, TMB, TMB,and andSTOP STOP solutiontotocome solution come to toroom room temperature. The temperature. The1x1xELISA ELISA buffer buffer included included in kit in kit waswas made made and loaded and loaded in reservoir.. in the the reservoir.. Media Media

from the plates from the plates was was discarded, discarded, the the ELISA ELISAplate platetapped tapped hard hard on on paper paper towel, towel, andand washed washed 4x 200 4x 200

µl1d ELISA wash ELISA wash bufferusing buffer using a a platewasher. plate washer.

One-hundred

[0938] One-hundred

[0938] (100) (100) µL/well of of

[L/well mouse mouse AR detection Ab Ab AR detection was was added; the the added; plates plates were were

covered andshaken covered and shakenat at37°C 37°C forfor 1 hour; 1 hour; media media was discarded was discarded from from the the plates, plates, the plates the plates were were

tapped on tapped on aa paper paper towel, towel, washed washed4x4xwith with200 200µL ELISA L ELISA wash wash buffer buffer with with a plate a plate washer. washer.

[0939] One-hundred

[0939] One-hundred (100) [L/well (100) µL/well of anti-mouse of anti-mouse - HRP conjugated - HRP conjugated Ab (comes Ab (comes with with the kit) the kit) was added; was added;the theplates plates were werecovered covered andand shaken shaken at 37°C at 37°C forminutes; for 30 30 minutes; the reagent the TMB TMB reagent was was allowed toto come allowed cometo toroom room temperature; temperature; the media the media was discard was discard from from the the the plate, plate, the plates plates were were tapped on tapped on paper papertowl, towl, washed washed4x4x with with 200200 L ELISA µL of of ELISA wash wash buffer; buffer; the plates the plates were were tapped tapped the the plates on plates paper towl. on paper towl. One-hundred One-hundred (100) (100) µL ofLTMB of was TMB wasandadded added and the the plates platesforshaken shaken 2 for 2 minutes -- while minutes while watching watching for for color color development. development. One-hundred (100) µLL Stop One-hundred (100) Stop solution solution was was added when added whenlight lightblue blue color color developed. developed. Plates Plates were wereshaken shakenand and readatat450 read 450nM. nM.

[0940] Progression

[0940] Progression of prostate of prostate cancer cancer in patients in patients treated treated withwith anti-androgen anti-androgen therapy therapy usually usually

involves one one of ofseveral several mechanisms mechanisms of of enhanced enhanced Androgen Androgen Receptor Receptor (AR) signaling, (AR) signaling, including including

increased increased intratumoral intratumoral androgen synthesis, increased androgen synthesis, increased AR expression and AR expression and ARAR mutations. mutations.

PROTACs PROTACs (PROteolysis (PROteolysis TArgeting TArgeting Chimera), Chimera), which which use use bi-functional bi-functional molecules molecules that that simultaneously binda target simultaneously bind a targetof of choice choice and and an E3anligase, E3 ligase, cause cause ubiquitination ubiquitination via induced via induced

proximity and proximity anddegradation degradationofofthe thetargeted, targeted, pathological pathological protein. protein. As Asopposed opposedtototraditional traditional target target inhibition, inhibition, which is a competitive which is process, degradation competitive process, degradationisisaaprogressive progressiveprocess. process.AsAssuch, such,it itisis less susceptibletotoincreases less susceptible increases in endogenous in endogenous ligand,ligand, target expression, target expression, or in or mutations mutations in the target. the target.

Thus, this Thus, this technology technologyappears appearstotobebeideal idealforforaddressing addressing thethe mechanisms mechanisms of ARof AR resistance resistance in in patients with patients with prostate prostate cancer. cancer. Data Data was analyzed and was analyzed andplotted plotted using using GraphPad GraphPad Prism Prism software. software.

[0941] 5. 5.

[0941] BRaf BRaf Protein In In Protein Vitro Vitro Degradation Degradation Assay Assay (A375 (A375 cells) Table 77 cells)ofof Table A375

[0942] A375

[0942] cells cells were were culturedininATCC cultured ATCC DMEM+10%FBS DMEM+10%FBS in 12 in 12 well well plates, plates, and treated and treated

with indicated with indicated compound compound from from Tables Tables 1-411-41 or 0.1% or 0.1% DMSO control DMSO vehicle vehicle for control for 16Cells 16 hours. hours. Cells were harvested were harvestedininCell CellSignaling Signalinglysis lysisbuffer buffer(Cat# (Cat#9803) 9803) with with thethe addition addition of of Roche Roche protease protease

inhibitor inhibitor tablets tablets(Cat# (Cat#11873580001), andlysates 11873580001), and lysates clarified clarified by by microcentrifugation. microcentrifugation. Proteins Proteins were were

401 separated by by SDS-PAGE, SDS-PAGE, and transferred onto nitrocellulose membranes using an using an Invitrogen 2023248067 10 Oct 2023 and transferred onto nitrocellulose membranes Invitrogen iBlot iBlot system. Immunoblotting was system. Immunoblotting was performed for BRaf performed for (Santa Cruz BRaf (Santa Cat# 9002), Cruz Cat# CRAF(BD 9002), CRAF (BD Cat#610151), andpErk Cat#610151), and pErk (CellSignaling (Cell Signaling Cat#9106). Cat#9106). GAPDH GAPDH (Cell Signaling (Cell Signaling Cat#2118) Cat#2118) was usedwas used as as aa loading loading control. control.Quantification Quantificationwas was carried carried out outusing usingthe theBioRad BioRad Image Lab5 5software. Image Lab software.

[0943] 6. 6.

[0943] BRaf BRaf In-Cell In-Cell Western Western Cellular Cellular Degradation Degradation Assay Assay (A375 (A375 cells) Table77 cells)ofofTable

[0944] A375

[0944] A375 cells cells were were cultured ATCC culturedininATCC DMEM+10% FBS in FBS DMEM+10% in 96-well 96-well plates,plates, and treated and treated

with indicated with indicated compounds compounds from from Tables-43 Tables-43 or 0.1% or 0.1% DMSO control DMSO vehicle vehicle for control for 72Cells 72 hours. hours. Cells were washed were washedwith with PBSPBS 1x, 1x, and and affixed affixed to plate to plate using using 4% in 4% PFA PFA in phosphate phosphate buffered buffered saline saline for for 15 minutes; washed 15 minutes; IX and washed 1X andpermeabilized permeabilized using using 0.1% 0.1%Triton-X-100 Triton-X-100 ininPBS PBSfor for5 5minutes; minutes; washed 1X washed IXand andblocked blocked with with LICOR LICOR blocker(Cat.# blocker (Cat.#927-50000) 927-50000) for for 11 hour. hour. Cells Cells were were then then incubated incubated with with B-Raf antibody (Santa B-Raf antibody (Santa Cruz Cruz Cat# Cat#9002, 9002,Santa SantaCruz CruzCat#528) Cat#528) andand tubulin tubulin

antibody (Sigma antibody (Sigma#T6074) #T6074) in in LICOR LICOR blocker blocker forhours. for 18 18 hours. CellsCells were were washed washed 3x to 3x prior prior to adding adding

secondary antibodies(LICOR secondary antibodies (LICORcat cat #926-32210 #926-32210 and 926-68071) and 926-68071) and incubated and incubated for 1Cells for 1 hour. hour. Cells were washed were washed3x3xandand imaged imaged using using LICOR LICOR Odyssey Odyssey Software. Software.

[0945] 7. 7.

[0945] BRD4 BRD4 Western Western Protocol Protocol for Table for Table 8 8

[0946] 22Rv-1

[0946] 22Rv-1 or VCaP were were cells cells or VCaP purchased purchased from and from ATCC ATCC and cultured cultured in Dulbecco's in Dulbecco's

Modified Eagle's Modified Eagle'sMedium Medium (ATCC), supplemented with (ATCC), supplemented with 10% FBS (ATCC) 10% FBS (ATCC) and and Penicillin/Streptomycin (Life Penicillin/Streptomycin (Life Technologies). Technologies). DMSO controlandand DMSO control compound compound treatments treatments

(0.003pM, 0.01pM, (0.003µM, 0.01µM, 0.03 0.03 pM 0.1µM) µM and and 0.1gM) were performed were performed in plates in 12-well 12-wellfor plates for 16Cells 16 hours. hours. Cells were harvested, were harvested, and and lysed lysedinin RIPA RIPAbuffer buffer(50mM (50mM Tris TrispH8, pH8, 150mM NaCl,1%1% 150mM NaCl, Tx-100,0.1% Tx-100, 0.1% SDS, 0.5% SDS, 0.5%sodium sodium deoxycholate)supplemented deoxycholate) supplemented with with protease protease andand phosphatase phosphatase inhibitors. inhibitors.

Lysates were Lysates wereclarified clarified atat 16,000g 16,000gforfor10 10 minutes, minutes, and and protein protein concentration concentration was determined. was determined.

Equal amountofofprotein Equal amount protein (20µg) (20pg)was wassubjected subjectedtotoSDS-PAGE SDS-PAGE analysis analysis and followed and followed by by immunoblotting according to immunoblotting according to standard standard protocols. protocols. The The antibodies antibodies used used were wereBRD4 BRD4 (Cell (Cell

Signaling #13440), Signaling #13440), and andActin Actin (Sigma (Sigma #5441). #5441). Detection Detection reagents reagents were Clarity were Clarity Western Western ECL ECL substrate substrate (Bio-rad (Bio-rad #170-5060). #170-5060).

402

Oct 2023

[0947]

[0947] Table 1. Table Exemplary Estrogen 1. Exemplary ReceptorPROTACs Estrogen Receptor PROTACs

Ex. Ex. Chemical Structure Chemical Structure Name Name General General # # Synthetic Synthetic

2023248067 10 Method Method 1 1 3-{5-[4-(5-{4-[(1S,2R)-6- 3-{5-[4-(5-{4-[(1S,2R)-6- A-2, A-8 A-2, A-8 O O hydroxy-2-phenyl-1,2,3,4- NH hydroxy-2-phenyl-1,2,3,4 NH tetrahydronaphthalen-1- N O tetrahydronaphthalen-1 N O yl]phenoxy}pentyl)piperazin- N yl]phenoxy}pentyl)piperazin N 1-yl]-7-methoxy-1-oxo-2,3- 1-yl]-7-methoxy-1-oxo-2,3 HO Ho O O N N dihydro-1H-isoindol-2 dihydro-1H-isoindol-2- yl}piperidine-2,6-dione yl}piperidine-2,6-dione

2 2 HO O 3-(5-(4-(5-(4-((1R, 2S)-6- 3-(5-(4-(5-(4-(1R, 2S)-6- A-2, A-8 A-2, A-8 HO hydroxy-2-phenyl-1,2,3,4- hydroxy-2-phenyl-1,2,3,4- Exp. Exp. O tetrahydronaphthalen-1- tetrahydronaphthalen-1- Procedure Procedure ~ O N N \ / N N NH NH yl)phenoxy)pentyl)piperazin- yl)phenoxy)pentyl)piperazin- included included N O 1 oxoisoindolin-2 yl) 7-methoxy-1I 1-yl)-7-methoxy-1- O oxoisoindolin-2- yl)piperidine-2,6-dione yl)piperidine-2,6-dione

3 3 0 O 0 3-[5-[4-[5-[4-[(1R,2S)-6- 3-[5-[4-[5-[4-[(1R,2S)-6- A-4, A-8 A-4, A-8 hydroxy-2-phenyl-tetralin-1- tetralin-1- Exp. Exp. HO NH NH hydroxy-2-phenyl- Ho Procedure 0 f NN N O yl] y1] Procedure O O phenoxy]pentyl]piperazin-1- phenoxy]pentyl]piperazin-1- included included N N O yl]-4-methoxy-1-oxo yl]-4-methoxy-1-oxo- isoindolin-2-yl]piperidine - isoindolin-2-y1]piperidine 2,6-dione 2,6-dione

403

Oct 2023 4 4 0 O 03-[5-[4-[5-[4-[(1R,2S)-6- 3-[5-[4-[5-[4-[(1R,2S)-6- A-4, A-5, A-4, A-5, A-8 A-8 HO Ho NH hydroxy-2-phenyl- tetralin-1 hydroxy-2-phenyl-tetralin-1- - 0 O N N x N t N NH 7yl]phenoxy]pentyl]piperazin yl]phenoxy]pentyl]piperazin- 1-yl]-4-[2-(2- N N 0methoxyethoxy)ethoxy]-1I methoxyethoxy)ethoxy]-1-

2023248067 10 la z foxo-isoindolin-2 oxo-isoindolin-2- 0 O yl]piperidine-2,6-dione yl]piperidine-2,6-dione

5 5 r:: N- 3 -(5 -14- [(1-14- [(1 S,2R)-6- 3-(5-{4-[(1-{4-[(1S,2R)-6- A-4, A-4, A-5, A-5, A A- N HO N NN 1,2,3,4- hydroxy-2-phenyl-1,2,3,4- hydroxy-2-phenyl- 12 12 Ho N tetrahydronaphthalen-1I tetrahydronaphthalen-1- yl]phenyl} piperidin-4- O O 0 yl)methyl]piperazin-1-yll-4 yl)methyl]piperazin-1-yl}-4- N N 0 o [2-(2

[2-(2-

0 NH methoxyethoxy)ethoxy]-1I methoxyethoxy)ethoxy]-1- O NH NH oxo-2,3-dihydro-1H oxo-2,3-dihydro-1H-

o 0 isoindol-2-yl)piperidine-2,6- isoindol-2-yl)piperidine-2,6 O dione

6 6 N3-(5-{4-[1-{4-[(R,2S)-6- 3-(5-{4-[(1-{4-[(1R,2S)-6- A-4, A-4, A-5, A-5, A A- N HO N Nhydroxy-2-phenyl- 1,2,3,4- hydroxy-2-phenyl-1,2,3,4- 12 12 Ho N -~ N tetrahydronaphthalen-1I tetrahydronaphthalen-1-

0 0 yl]phenyllpiperidin-4 yl]phenyl} piperidin-4-

yl)methyl]piperazin-1-yl}-4- N0 ~ yl)methyl]piperazin-1I-yl}1-4 N N 0 [2-(2

[2-(2-

N H methoxyethoxy)ethoxy]-1I methoxyethoxy)ethoxy]-1- O NH NH oxo-2,3-dihydro-1H oxo-2,3-dihydro-1H- isoindol-2-yl)piperidine-2,6- 0o isoindol-2-yl)piperidine-2,6 O dione

404

7 f )(3 S)-3 -(5 -{2-[4-(4-{4- (3S)-3-(5-{2-[4-(4-{4- A-5, A-5, A-9 A-9

/N N _N [(I S,2R)-6-hydroxy-2

[(1S,2R)-6-hydroxy-2- N phenyl- 1,2,3,4 phenyl-1,2,3,4-

O 0 tetrahydronaphthalen-1I tetrahydronaphthalen-1- /\ Iyl]phenoxy Ibutyl)- 1,4 yl]phenoxy}butyl)-1,4-

O diazepan-1-yl]ethyl}-4-[2-(2- 0 methoxyethoxy)ethoxy]-1I methoxyethoxy)ethoxy]-1- H O\ N O oxo-2,3-dihydro-1H Ho oxo-2,3-dihydro-1H-

2023248067 on ""\.'I / NH NH isoindol-2-yl)piperidine-2,6 isoindol-2-yl)piperidine-2,6-

dione dione 0 O 8 8 (3S)-3-[5-[2-[4-[4-[4- (3S)-3-[5-[2-[4-[4-[4- A-5, A-5, A-9 A-9

N N [(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- O-J/--N phenyl-tetralin-1I phenyl-tetralin-1-

O 0 yl]phenoxy]butyl]- 1,4 yl]phenoxy]butyl]-1,4-

/\Idiazepan-1I-yl] ethyl] -4- [2-(2 diazepan-1-yl]ethyl]-4-[2-(2- N 0 methoxyethoxy)ethoxy]-1I methoxyethoxy)ethoxy]-1- oxo-isoindolin-2- oxo-isoindolin-2 HON,0 HO\/ N yl]piperidine-2,6-dione HO yl]piperidine-2,6-dione NH NH

0 O 9 9 HO \ -5-4[-l1V+l S,2R)-6- 3-(5-{4-[2-(1-{4-[(1S,2R)-6- A-2, A-2, A- 13 A-13 HO hydroxy-2-phenyl-1,2,3,4- hydroxy-2-phenyl- 1,2,3,4 - O 0 tetrahydronaphthalen-1I ~ tetrahydronaphthalen-1- - - Nyl]phenyllpiperidin-4 N yl]phenyl}piperidin-4- N \ / N N NHyl)ethyl]piperazin-1I-yl} NH -7 yl)ethyl]piperazin-1-yl}-7- 0 methoxy-1I-oxo-2,3 -dihydro methoxy-1-oxo-2,3-dihydro- 1H-isoindol-2-yl)piperidine 1H-isoindol-2-yl)piperidine- 2,6-dione 2,6-dione

405

Oct 2023 10 10 HO \03-[5-[4-[2-[1-[4-[(1R,2S)-6- 3-[5-[4-[2-[1-[4-[(1R,2S)-6- A-2, A-2, A- 13 A-13 Ho 10 ~ hydroxy-2-phenyl-tetralin-1I hydroxy-2-phenyl-tetralin-1- 0 yl]phenyl]-4 yl]phenyl]-4-

N- N ~ NN \/piperidyl]ethyl]piperazin-1I piperidyl]ethyl]piperazin-1- \ /N N N NHyl]-7-methoxy-1-oxo NH yl]-7-methoxy-1-oxo-

2023248067 10 0 isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine 2,6-dione 2,6-dione

11 11 N3-[5-[4-[[1-[4 -[(1R,2S)-6- 3-[5-[4-[[1-[4 -[(1R,2S)-6- A-4, A-4, A-12 A-12 N HO Ho N N N N hydroxy-2-phenyl-tetralin-1 hydroxy-2-phenyl-tetralin-1- yl]phenyl]-4 yl]phenyl]-4-

0 0 piperidyl]methyl]piperazin-1I piperidyl]methyl]piperazin-1- o O yl] -4-(2-methoxyethoxy)-1I yl]-4-(2-methoxyethoxy)-1- N N 0 O oxo-isoindolin-2 oxo-isoindolin-2- yl]piperidine-2,6-dione yl]piperidine-2,6-dione NH O NH

0 O 12 12 03-[5-[4-[4,4-difluoro-5-[4- 3-[5-[4-[4,4-difluoro-5-[4- A-2, A-2, A- 1 A-10 O 0 [(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- HO Ho O FEF o phenyl-tetralin-1I phenyl-tetralin-1- F F O r-\N \ N-, N NH NH yl]phenoxy]pentyl]piperazin yl]phenoxy]pentyl]piperazin- N N 0 11-yl]-7-methoxy-1-oxo- -yl]-7-methoxy-1I-oxo N O isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine-

2,6-dione 2,6-dione

406

Oct 2023 13 13 1\0(3R)-3-[5-[4-[5-[4-[(1R,2S)- (3R)-3-[5-[4-[5-[4-[(1R,2S)- A-2, A-2, A-8 A-8 o0 6-hydroxy-2-phenyl-tetralin 6-hydroxy-2-phenyl-tetralin- 1- HO NHI NH Ho O NN"" 67 N N3 O yl]phenoxy]pentyl]piperazin- yl]phenoxy]pentyl]piperazin IN N 11-yl]-7-methoxy-1-oxo- -yl]-7-methoxy-1I-oxo 2023248067 10 isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine- 2,6-dione 2,6-dione

14 14 "\a (3S)-3-[5-[4-[5-[4-[(1R,2S)- (3S)-3-[5-[4-[5-[4-[(1R,2S)- A-2, A-2, A-8 A-8 O 6-hydroxy-2-phenyl-tetralin o O0 6-hydroxy-2-phenyl-tetralin- 1- HO NHI NH Ho yl]phenoxy]penty|]piperazin- ) N No 0 yl]phenoxy]pentyl]piperazin o I I NN -yl--etoy ~~N O I Ioo 1-yl]-7-methoxy-1-oxo- 1y]7mtoy1oo N isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine- 2,6-dione 2,6-dione

15 15 N 3-[5-[4-[[1-[4-[(1R,2S)-6- 3-[5-[4-[[1-[4-[(1R,2S)-6- A-2, A-2, A-12 A-12 N HO 0 N hydroxy-2-phenyl -tetralin-1 hydroxy-2-phenyl -tetralin-1- Ho N N o yl]phenyl]-4 yl]phenyl]-4-

0 ~ piperidyl]methyl]piperazin-1I piperidyl]methyl]piperazin-1- O yl]-7-methoxy-1-oxo yl]-7-methoxy-1-oxo- N N 0 o isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine 2,6-dione 2,6-dione ICNH NH

0__ _ _ _ _ _ _ _ _ _ _ O

407

16 3-[5-[4-[5-[4-[(1R,2S)-2-(4- 3-[5-[4-[5-[4-[(1R,2S)-2-(4- A-2, A-2, A-IlI A-11 0 ~fluorophenyl)-6- hydroxy fluorophenyl)-6-hydroxy- O N N N N \0 tetralin-1I-yl]phenoxy] tetralin-1-yl]phenoxy]- O VJ \ 1,2,3,3a,4,5,6,6a 1,2,3,3a,4,5,6,6a-

N, N octahydropentalen-2 octahydropentalen-2- HO yl]piperazin-l-yl]-7 yl]piperazin-1-yl]-7- Ho Oi .N IZ methoxy-1I-oxo-isoindolin-2 methoxy-1-oxo-isoindolin-2- N O F FH H yl]piperidine-2,6-dione yl]piperidine-2,6-dione

17 17 0 0 3-[5-[4-[[6-[[4-[(1R,2S)-6- 3-[5-[4-[[6-[[4-[(1R,2S)-6- A-16 A-16 O 2023248067

O O hydroxy-2-phenyl-tetralin-1- NH hydroxy-2-phenyl-tetralin-1I NH yl]phenoxy]methy[]-2- N_0 yl]phenoxy]methyl]-2 N pyridyl]methyl]piperazin-1- -~N pyridyl]methyl]piperazin-1I N Nryl]-7-methoxy-1-oxo yl]-7-methoxy-1-oxo- HO0, Ho H0~oO N isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine- N 2,6-dione 2,6-dione

18 18 F F r N 3-(5-{4-[(1-{2,6-difluoro-4- 3-(5-{4-[(1-{2,6-difluoro-4- A-i, A-1, A-2 A-2 N HO N N 0, [(I1S,2R)-6-hydroxy-2

[(1S,2R)-6-hydroxy-2- Ho N N O phenyl- 1,2,3,4 phenyl-1,2,3,4- i'll F tetrahydronaphthalen-1I tetrahydronaphthalen-1- F F 0O yl]phenyllpiperidin-4 yl]phenyl} piperidin-4- N N 0 O yl)methyl]piperazin-1I-yl}1-7 yl)methyl]piperazin-1-yl}-7- methoxy-1I-oxo-2,3 -dihydro methoxy-1-oxo-2,3-dihydro- NH NH 1H-isoindol-2-yl)piperidine 1H-isoindol-2-yl)piperidine- L 2,6-dione 2,6-dione 0__ O _ _ _ _ _ _ _ _ _ _

408

Oct 2023 19 19 F F r N 3-(5-(4-((1-(2,6-difluoro-4- 3-(5-(4-((1-(2,6-difluoro-4- A-i, A-1, A-2, A-8 A-8 N HO0 N N ((1R,2S)-6-hydroxy-2 ((1R,2S)-6-hydroxy-2- Ho N N O phenyl- 1,2,3,4 phenyl-1,2,3,4-

F 0 tetrahydronaphthalen-1I tetrahydronaphthalen-1- F F O yl)phenyl)piperidin-4 yl)phenyl)piperidin-4-

2023248067 10 N N 0 O yl)methyl)piperazin-1I-yl)-7 yl)methyl)piperazin-1-yl)-7- methoxy-1I-oxoisoindolin-2 methoxy-1-oxoisoindolin-2- NH yl)piperidine-2,6-dione yl)piperidine-2,6-dione INH 0 O 20 20 F F 3 -[7-(difluoromethoxy)-5 -[4- 3-[7-(difluoromethoxy)-5-[4- A-3, A-3, A-8 A-8

HO Ho FF [5-[4-[(1R,2S)-6- hydroxy-2

[5-[4-[(1R,2S)-6- hydroxy-2-

O 0 phenyl-tetralin-1- phenyl-tetralin-1I O 0 yl]phenoxy]pentyl]piperazin yl]phenoxy]pentyl]piperazin- O 1-yl]-1-oxo-isoindolin-2-

N N N\~JN \I N Nyl]piperidine-2,6-dione yl]piperidine-2,6-dione

IZ N O o 0 H H 21 21 F F 3-[5-[4-[4,4-difluoro-5-[4- 3-[5-[4-[4,4-difluoro-5-[4- A-6, A-6, A-10 A-10

[(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- F F O 0 phenyl-tetralin-1I phenyl-tetralin-1- O O yl]phenoxy]pentyl]piperazin yl]phenoxy]pentyl]piperazin- HO Ho F F N N1-yl]-7-(difluoromethoxy)-1I NH 1-yl]-7-(difluoromethoxy)-1- -0 O -. F F fN N NN 6N 0 N oxo-isoindolin-2 o oxo-isoindolin-2- N yl]piperidine-2,6-dione yl]piperidine-2,6-dione

409

22 F F 3 -[7-(difluoromethoxy)-5 -[4- 3-[7-(difluoromethoxy)-5-[4- A-2, A-2, A- 13 A-13 F4 F [2-[1-[4-[(1R,2S) -6

[2-[1-[4-[(1R,2S) -6- HO Ho 0 O hydroxy-2-phenyl-tetralin-1I hydroxy-2-phenyl-tetralin-1- \N00 O yl]phenyl]-4 yl]phenyl]-4- O -- NNNN \ -dN \/ piperidyl]Jethyl]piperazin-1- piperidyl]ethyl]piperazin-1I N \/ N N NHyl]-1I-oxo-isoindolin-2 NH yl]-1-oxo-isoindolin-2-

0 yl]piperidine-2,6-dione yl]piperidine-2,6-dione O 2023248067

23 23 FF 3-[7-(difluoromethoxy)-5-[4- 3-[7-(difluoromethoxy)-5-[4- A-3, A-14 A-3, A-14 F [2-[3-[4-[(1R,2S)-6-hydroxy

[2-[3-[4-[(1R,2S)-6-hydroxy- 0 O 0 O 2-phenyl-tetralin-1I 2-phenyl-tetralin-1-

/ N yl]phenoxy] cyclobutyl] ethyl] yl]phenoxy]cyclobutyl]ethy]] N N O 0piperazin-1I-yl]-1I-oxo piperazin-1-yl]-1-oxo- HO/\ HO Nisoindolin-2-yl]piperidine isoindolin-2-yl]piperidine- Ho NX,. L2,6-dione 2,6-dione

o O N ZI N 0 H H O 24 24 H H 3-[5-[4-[2-[1-[2-fluoro-4- 3-[5-[4-[2-[1-[2-fluoro-4- A-2, A-2, A-12 A-12 HO 0 O N N [(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- Ho phenyl-tetralin-1I-yl]phenyl] phenyl-tetralin-1-yl]phenyl]-

F F N 4-piperidyl]ethyl]piperazin 4-piperidyl]ethyl]piperazin- N I 1-yl]-7-methoxy-1I-oxo 1-yl]-7-methoxy-1-oxo- N NN N \/ aisoindolin-2-yl]piperidine isoindolin-2-yl]piperidine- N N N \-IO -C to2,6-dione 2,6-dione 0

410

25 HO 0 3{j7[4-(5{j44(R,2S)-6 A-7,A-8 2023248067 10 Oct 2023

25 Ho 3-{7-[4-(5-{4-[(1R,2S)-6- A-7, A-8 NN N N O hydroxy-2-phenyl- 1,2,3,4 hydroxy-2-phenyl-1,2,3,4- N N 'sN tetrahydronaphthalen-1I tetrahydronaphthalen-1- N N yl]phenoxylpentyl)piperazin N yl]phenoxy}pentyl)piperazin-

O 0 N IZ 0 -l--oo2,H 1-yl]-3-oxo-2H,3H- O N H [1,2,4]triazolo[4,3-a]pyridin

[1,2,4]triazolo[4,3-a]pyridin- H 2-yllpiperidine-2,6-dione 2-yl}piperidine-2,6-dione

26 0 3-[5-[4-[2-[3-[4-[(1R,2S)-6- 3-[5-[4-[2-[3-[4-[(1R,2S)-6- A-17 26 A-17 0-< O 0 hydroxy-2-phenyl-tetralin-1I O O hydroxy-2-phenyl-tetralin-1- /\N N N N 0yl]phenoxy]cyclobutyl]-2 O yl]phenoxy]cyclobutyl]-2- ~ \/oxo-ethyl]piperazin-lI-yl] -7 oxo-ethyl]piperazin-1-yl]-7- HO Ho ' N, N methoxy-1I-oxo-isoindolin-2 methoxy-1-oxo-isoindolin-2- yl]piperidine-2,6-dione yl]piperidine-2,6-dione o ZIN 0 O N H H 27 27 FF 3 -[7-(difluoromethoxy)-5 -[4- 3-[7-(difluoromethoxy)-5-[4- A-3, A-3, A-15 A-15 OH OH F [2- [1 -hydroxy-3 -[4+[1 R,2S)

[2-[1-hydroxy-3-[4-[(1R,2S)- o O 0 O 6-hydroxy-2-phenyl-tetralin 6-hydroxy-2-phenyl-tetralin- 1- 0 1 N N ~\/yl]phenoxy] O cyclobutyl] ethyl] yl]phenoxy]cyclobutyl]ethy[]

HO Ho -\ Nr piperazin-lI-yl]-lI-oxo piperazin-1-yl]-1-oxo- N isoindolin-2-yl]piperidine isoindolin-2-yl]piperidine X,. L2,6-dione 2,6-dione 0 O NN IZ 0 H O H 28 28 OH OH 3-[5-[4-[2-[ 1-hydroxy-3-[4- 3-[5-[4-[2-[1-hydroxy-3-[4- A-2, A-2, A-i15 A-15 O 00 o [(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- /\NNN N N O phenyl-tetralin-1I phenyl-tetralin-1-

yl]phenoxy]cyclobutylJethy] HO HO \ N N piperazin-1I-yl]-7-methoxy-1I piperazin-1-yl]-7-methoxy-1- oxo-isoindolin-2 oxo-isoindolin-2- oN ZI yl]piperidine-2,6-dione yl]piperidine-2,6-dione O N O HH

411

29 N3-(6'-{4-[1-{4-+(R,2S)-6- 3-(6'-{4-[(1-{4-[(1R,2S)-6- A-6, A-6, A-12 A-12 N N N ~ hydroxy-2-phenyl- 1,2,3,4 hydroxy-2-phenyl-1,2,3,4- N N tetrahydronaphthalen-1I tetrahydronaphthalen-1- HO 0 Ho 0 yl]phenyllpiperidin-4 yl]phenyl} piperidin-4- O NN 0 O yl)methyl]piperazin-lI-yl} -3' yl)methyl]piperazin-1-yl}-3'- oxo-2',3' oxo-2',3'- NH NH dihydrospiro~cyclopropane dihydrospiro[cyclopropane-

9zl- (41,1'-isoindole]-2' 1,1'-isoindole]-2'- 0 O yl)piperidine-2,6-dione yl)piperidine-2,6-dione 30 N3-[6'-[4-[[1-[2-fluoro-4- A-6, A-6, A-12 2023248067

30 3-[6'-[4-[[1-[2-fluoro-4- A-12 HO HO N N [(1R,2S)-6-hydroxy-2

[(1R,2S)-6-hydroxy-2- I N N phenyl-tetralin-1I-yl]phenyl] phenyl-tetralin-1-yl]phenyl]-

F F I~.4-piperidyl]methyl]piperazin 4-piperidyl]methyl]piperazin- aO 1-yl]-3'-oxo-spiro 1-yl]-3'-oxo-spiro

[cyclopropane-1,1'-

[cyclopropane-1,1' NN 0 O isoindoline]-2'-yl]piperidine isoindoline]-2'-yl]piperidine-

H NNH 2,6-dione 2,6-dione

0 o 31 31 NA-i, A-1, A-2 A-2 N HO0 Ho N N N0 N O

- ~- 0 O N N 0 O

NH O

412

Receptor PROTACs Androgen Receptor Exemplary Androgen 10 Oct 2023

[0948]

[0948] 2.Exemplary Table 2. Table PROTACs

Ex. ChemicalStructure Name General General Ex. Chemical Structure Name # scheme rac-N-((1r,4r)-4-(3-chloro-4 rac-N-(1r,4r)-4-(3-chloro-4- cyanophenoxy)cyclohexyl) cyanophenoxy)cyclohexyl)- N O O 6-(4-((4-(2-(2,6 6-(4-((4-(2-(2,6-

N N N NH NH dioxopiperidin-3-yl)-7- dioxopiperidin-3-yl)-7- Exp. Exp. H H 32 32 N N NO N NN O methoxy-1,3- methoxy-1,3- procedure procedure N N 2023248067

1IPJN O O O dioxoisoindolin-5- dioxoisoindolin-5- yl)piperazin-1 yl)piperazin-1- provided provided

c1 CI 0 O yl)methyl)piperidin-1 yl)methyl)piperidin-1- yl)pyridazine-3-carboxamide yl)pyridazine-3-carboxamide rac-N-((1r,3r)-3-(3-chloro-4 rac-N-(1r,3r)-3-(3-chloro-4- cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- tetramethylcyclobutyl)-2-(4- tetramethylcyclobutyl)-2-(4 N N 0 O NH ((4-(2-(2,6-dioxopiperidin-3- ((4-(2-(2,6-dioxopiperidin-3 N N N N N N NH N ZI yl)-7-methoxy-1,3- 33 33 NH H N N O o yl)-7-methoxy-1,3- B-1 B-2 B-1, B-2 N N dioxoisoindolin-5 dioxoisoindolin-5-

o yl)piperazin-1- C0 CI 0 0O yl)piperazin-1 O O yl)methyl)piperidin-1 yl)methyDpiperidin-1- yl)pyrimidine-5 yl)pyrimidine-5- carboxamide carboxamide rac-N-((1r,3r)-3-(3-chloro-4 rac-N-(1r,3r)-3-(3-chloro-4- cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- cl CI O O O tetramethylcyclobutyl)-4-(4 tetramethylcyclobutyl)-4-(4- O ((4-(2'-(2,6-dioxopiperidin- Synthesis Synthesis HN, NH NH ((4-(2'-(2,6-dioxopiperidin- 34 NN O 3-yl)-3'- 3-yl)-3'- described in described in 34 N N N N N oxospiro[cyclopropane-1,1'- oxospiro[cyclopropane-1,1'- detail detail N N N isoindolin]-6'-yl)piperazin-1 isoindolin]-6'-yl)piperazin-1-

yl)methyl)piperidin-1 yl)methyl)piperidin-1- yl)benzamide yl)benzamide

413

Oct 2023 rac-N-((1r,4r)-4-(3-chloro-4 rac-N-(1r,4r)-4-(3-chloro-4- cyanophenoxy)cyclohexyl) cyanophenoxy)cyclohexyl)- N N 5-(4-((((1r,3r)-3-((2'-(2,6- 5-(4-(((1r,3r)-3-(2'-(2,6- Synthesized Synthesized N N." 0 N dioxopiperidin-3-yl)-3'- following theO 35 35N O o NN ~~ N O N o NH dioxopiperidin-3-yl)-3'-

oxospiro[cyclopropane-1,1'- oxospiro[cyclopropane-1,1'- following the route route CI .NH N N NH CI 0 isoindolin]-5'- isoindolin]-5'- described for described for 2023248067 10

Oo NH O yl)oxy)cyclobutyl)(isopropyl Ex. Comp. yl)oxy)cyclobutyl)(isopropyl Ex. 34 Comp. 34 )amino)methyl)piperidin-1 )amino)methyl)piperidin-1- yl)pyrazine-2-carboxamide yl)pyrazine-2-carboxamide rac-N-((1r,4r)-4-(3-chloro-4 rac-N-(1r,4r)-4-(3-chloro-4- cyanophenoxy)cyclohexyl) cyanophenoxy)cyclohexyl)- N 10 5-(4-((((lr,3r)-3-((2'-(2,6- 5-(4-((1r,3r)-3-((2'-(2,6- Synthesized Synthesized N O3 NN 36 36 O N N O N O O NH dioxopiperidin-3-yl)-3'- dioxopiperidin-3-yl)-3'-

oxospiro[cyclopropane-1,1'- oxospiro[cyclopropane-1,1'- following the following route route the CI N N NH NH isoindolin]-6'- isoindolin]-6'- described for described for O 0 4 o yl)oxy)cyclobutyl)(isopropyl yl)oxy)cyclobutyl)(isopropyl Ex. Comp. Ex. Comp. 3434 o )amino)methyl)piperidin-1 )amino)methyl)piperidin-1- yl)pyrazine-2-carboxamide yl)pyrazine-2-carboxamide rac-N-((1r,3r)-3-(3-chloro-4 rac-N-((1r,3r)-3-(3-chloro-4-

cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- tetramethylcyclobutyl)-5-(4- NN\N o O O tetramethylcyclobutyl)-5-(4- Synthesized Synthesized N o N N N .N N O , N O 0((((Ilr,3r)-3 NH NH -((2'-(2,6- ((((1r,3r)-3-((2'-(2,6- dioxopiperidin-3-yl)-3' dioxopiperidin-3-yl)-3'- following the following the 37 N route CI CI .%NH N oxospiro[cyclopropane-1,1'- o oxospiro[cyclopropane-1,1'- described for for 0 O NH isoindolin]-5'- isoindolin]-5 - described Ex. Comp. Ex. 34 Comp. 34 yl)oxy)cyclobutyl)(isopropyl yl)oxy)cyclobutyl)(isopropyl )amino)methyl)piperidin-1 )amino)methyl)piperidin-1- yl)pyrazine-2-carboxamide yl)pyrazine-2-carboxamide

414

rac-N-((1r,3r)-3-(3-chloro-4 rac-N-(1r,3r)-3-(3-chloro-4- cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- N N N tetramethylcyclobutyl)-5-(4- tetramethylcyclobutyl)-5-(4- Synthesized Synthesized N ((((1r,3r)-3-((2'-(2,6- N NNN N0 ((((1r,3r)-3-((2'-(2,6- following the 38 38 38 O N O /s"' O dioxopiperidin-3-yl)-3'- dioxopiperidin-3-yl)-3'- following the foroutete CI ,NH N N NH oxospiro[cyclopropane-1,l'- oxospiro[cyclopropane-1,1'- NH described for described for O 0- 0 O isoindolin]-6'- isoindolin]-6'- E.Cm.3 O Ex. Comp. 34 yl)oxy)cyclobutyl)(isopropyl yl)oxy)cyclobutyl)(isopropyl Ex.Comp.34 )amino)methyl)piperidin-1 )amino)methyl)piperidin-1- 2023248067

yl)pyrazine-2-carboxamide yl)pyrazine-2-carboxamide N N rac-N-((1r,3r)-3-(3-chloro-4 rac-N-(1r,3r)-3-(3-chloro-4- cyanophenoxy)-2,2,4,4- CI cyanophenoxy)-2,2,4,4 tetramethylcyclobutyl)-6-(4- tetramethylcyclobutyl)-6-(4- C-i and C-1 andExp. Exp. O1 \NH (5-((2-(2,6-dioxopiperidin-3- (5-((2-(2,6-dioxopiperidin-3- procedure procedure 46 46 O" NH N N N N yl)-1-oxo-1,2,3,4- yl)-1-oxo-1,2,3,4- provided as provided as O N N - tetrahydroisoquinolin-6- tetrahydroisoquinolin-6- well well O NH yl)oxy)pentyl)piperazin-1- yl)oxy)pentyl)piperazin-1 O O NH N O yl)nicotinamide yl)nicotinamide N O ZI H N rac-N-(1r,3r)-3-(3-chloro-4- rac-N-((lr,3r)-3-(3-chloro-4 0O N O O cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- N tetramethylcyclobutyl)-6-(4- tetramethylcyclobutyl)-6-(4- C-i and C-1 andExp. Exp. N O (5-((2-(2,6-dioxopiperidin-3- (5-((2-(2,6-dioxopiperidin-3- procedure procedure 47 47 N O N N yl)-1,3-dioxo-1,2,3,4- yl)-1,3-dioxo-1,2,3,4- provided as provided as N N N ZI N tetrahydroisoquinolin-6- tetrahydroisoquinolin-6- well well N N yl)oxy)pentyl)piperazin-1 yl)oxy)pentyl)piperazin-1- ci CI o'' O" O yl)nicotinamide yl)nicotinamide

415

0 0 rac-N-((l r,3r)-3 -(3 -chloro-4 10 Oct 2023

rac-N-(1r,3r)-3-(3-chloro-4-

N NH NH cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- N tetramethylcyclobutyl)-6-(4- N tetramethylcyclobutyl)-6-(4 N O (5-((2-(2,6-dioxopiperidin-3- 48 48 N~)(5-((2-(2,6-dioxopiperidin-3- Exp procedure Exp procedure 4NH N Iyl)-3-oxo-2,3-dihydro- yl)-3-oxo-2,3-dihydro- provided provided N ZI H NN N [1,2,4]triazolo[4,3-a]pyridin

[1,2,4]triazolo[4,3-a]pyridin-

CI O" ol 7-yl)oxy)pentyl)piperazin-1I 7-yl)oxy)pentyl)piperazin-1- 0:)T yl)nicotinamide yl)nicotinamide

2023248067 0 o rac-N-((l r,3r)-3 -(3 -chloro-4 rac-N-(1r,3r)-3-(3-chloro-4- - N NH cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- N o tetramethylcyclobutyl)-6-(4- N 0 .. N N tetramethylcyclobutyl)-6-(4 N (5-((2-(2,6-dioxopiperidin-3- 49 490 O (5-((2-(2,6-dioxopiperidin-3- Exp procedure Exp procedure 49 NH I~ yl)-3-oxo-2,3-dihydro- Nprovided yl)-3-oxo-2,3-dihydro- provided N HN

N N -:JN

[1,2,4]triazolo[4,3-a]pyridin

[1,2,4]triazolo[4,3-a]pyridin-

C 0 6-yl)oxy)pentyl)piperazin-1I 6-yl)oxy)pentyl)piperazin-1- CI o O yl)nicotinamide yl)nicotinamide rac-N-((1 r,3r)-3 -(3 -chloro-4 rac-N-(1r,3r)-3-(3-chloro-4-

a cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- O o 0 0 0tetramethylcyclobutyl)-4-(4 tetramethylcyclobutyl)-4-(4- o N NNk N ((4-(2-(2,6-dioxopiperidin-3-Eprcdr ((4-(2-(2,6-dioxopiperidin-3- Exp procedure 50 50 N IIH NN N 0N~ O yl)-3-oxo-2,3-dihydro- yl)-3-oxo-2,3-dihydro- xprcde N CI N NNN0 N NH [1,2,4]triazolo[4,3-a]pyridin-

[1,2,4]triazolo[4,3-a]pyridin- poie provided

N N 7-yl)piperazin-l 7-yl)piperazin-1- yl)methyl)piperidin-1I yl)methyl)piperidin-1- yl)benzamide yl)benzamide

416 oo 0o N-((l r,3 r)-3 -(3 -chloro-4 N-(1r,3r)-3-(3-chloro-4- N- N N NH H cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4- N O tetramethylcyclobutyl)-4-(4- N N N N tetramethylcyclobutyl)-4-(4 N 0 O (2-(4-(2-(2,6-dioxopiperidin- (2-(4-(2-(2,6-dioxopiperidin- C-3, C-4 C-3, and C-4 and 51 51 N 3-yl)-l,3-dioxo-2,3-dihydro- 3-yl)-1,3-dioxo-2,3-dihydro- Exp procedure Exp procedure N 1H-pyrrolo[3,4-c]pyridin-6- provided provided N HN N H-pyrrolo[3,4-c]pyridin-6- 1H ~yl)piperazin-1I yl)piperazin-1- -lr yl)ethyl)piperidin-1I yl)ethyl)piperidin-1-

CI CI ~ 0'ybezmd O O' 00 yl)benzamide 2023248067

N o 0 o0 N-(1r,3r)-3-(3-chloro-4- NH N-((l r,3 r)-3 -(3 -chloro-4 N-, N O N I cyanophenoxy)-2,2,4,4 cyanophenoxy)-2,2,4,4-

N N N N tetramethylcyclobutyl)-4-(4 tetramethylcyclobutyl)-4-(4-

N) 0 (2-(4-(6-(2,6-dioxopiperidin- (2-(4-(6-(2,6-dioxopiperidin- C-3, c-4 and C-3, c-4 and N o 52 52 N3-yl)-5,7-dioxo-6,7-dihydro- 3-yl)-5,7-dioxo-6,7-dihydro- exp exp procedure procedure N 5H-pyrrolo[3,4-d]pyrimidin- 5H-pyrrolo[3,4-d]pyrimidin- provided provided N- N NH 2-yl)piperazin-1 2-yl)piperazin-1- NH yl)ethyl)piperidin-1-

CI ________________________________________ O" O yl)benzamide yl)benzamide

417

3.Exemplary Table 3. Table BRaf PROTACs Exemplary BRaf Oct 2023

[0949]

[0949] PROTACs

Ex. Chemical Structure Name Synthetic Synthetic Ex. Chemical Structure Name # Scheme HO-N Ho N 2023248067 10 (E)-2-(2,6-dioxopiperidin-3-yl)-6-(4 (E)-2-(2,6-dioxopiperidin-3-yl)-6-(4- (4-(4-(1-(hydroxyimino)-2,3-dihydro (4-(4-(1-(hydroxyimino)-2,3-dihydro- 39 39 N N \N / O 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- D-1 D-1 N N N O pyrazol-1-yl)phenyl)piperazin-1-yl)- N.~ N pyrazol-1-yl)phenyl)piperazin-1-yl) N N NH 4-phenylisoindoline-1,3-dione 4-phenylisoindoline-1,3-dione NH N N 0 O O 0 0 o O HO-N HO-N \N N NH (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4 (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4- N N NH N \__Nj) (4-(4-(1-(hydroxyimino)-2,3-dihydro (4-(4-(1-(hydroxyimino)-2,3-dihydro- 40 40 N O O 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- 1H-inden-5-yl)-3-(pyridin-4-y1)-1H- D-1 D-1 N O N pyrazol-1-yl)phenyl)piperazin-1-yl) pyrazol-1-yl)phenyl)piperazin-1-yl)- 6-methylisoindoline-1,3-dione 6-methylisoindoline-1,3-dione

N N 0 O o NH NH N N o 0 N (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4- N(E)-2-(2,6-dioxopiperidin-3-yl)-5-(4 N N o O (4-(4-(1-(hydroxyimino)-2,3-dihydro- (4-(4-(1-(hydroxyimino)-2,3-dihydro- Custom Custom 41 41 -N 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- synthesis synthesis HO-N /ON\pyrazol-1-yl)phenyl)piperazin-1-yl)- N pyrazol-1-yl)phenyl)piperazin-1-yl)- provided provided -N N 4-phenylisoindoline-1,3-dione 4-phenylisoindoline-1,3-dione

N N

418

O 00 NH | N N 0 O N (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4- NN(E)-2-(2,6-dioxopiperidin-3-yl)-5-(4 N N o (4-(4-(1-(hydroxyimino)-2,3-dihydro- (4-(4-(1-(hydroxyimino)-2,3-dihydro- Custom Custom 42 42 -N 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- 1H-inden-5-yl)-3-(pyridin-4-yl)-1H- synthesis synthesis HO-N HO\ \NN Z-z pyrazol-1-yl)phenyl)piperazin-1-yl)- pyrazol-1-yl)phenyl)piperazin-1-yl)- provided provided -N N 4-methylisoindoline-1,3-dione 4-methylisoindoline-1,3-dione

2023248067

N N

419

Oct 2023 109501

[0950] Table 4. Table 4.Exemplary ExemplaryBRD4 BRD4 PROTACs PROTACs

Ex. Ex. # # Chemical Structure Chemical Structure Name Name O 0 2-((R)-4-(4-chlorophenyl)-2,3,9 2-(R)-4-(4-chlorophenyl)-2,3,9-

trimethyl-6H-thieno[3,2 trimethyl-6H-thieno[3,2- N-NN N N5 IZ a O0 O - N f][1,2,4]triazolo[4,3 ,, H H H ZI H f][1,2,4]triazolo[4,3- O 2023248067 10

4N N /N N0\- O N 0 a][1,4]diazepin-6-yl)-N-(4-(2-(2 a][1,4]diazepin-6-yl)-N-(4-(2-(2- 43 0-\ H ZI (S) ()(2-(2-((1 -oxo-2-((R)-6 (2-(2-((1-0xo-2-((R)-6- S O H N NN N oxopiperidin-3-yl)isoindolin-4 oxopiperidin-3-yl)isoindolin-4- CI CI - yl)amino)ethoxy)ethoxy)ethoxy)eth yl)amino)ethoxy)ethoxy)ethoxy)eth

\/ O 0 oxy)phenyl)acetamide oxy)phenyl)acetamide

2-(R)-4-(4-chlorophenyl)-2,3,9- H H 2-((R)-4-(4-chlorophenyl)-2,3,9 0 O N N 0 trimethyl-6H-thieno[3,2 trimethyl-6H-thieno[3,2- O sS N~~L N i f][1,2,4]triazolo[4,3 f][1,2,4]triazolo[4,3- N N N N N a] [1,4]diazepin-6-yl)-N-(4-(2-(2 a][1,4]diazepin-6-yl)-N-(4-(2-(2- N o 44 44 o 0~.O~N O OO (2-(2-((3-(2,6-dioxopiperidin-3-yl) (2-(2-(3-(2,6-dioxopiperidin-3-yl)- N A "'-,--- IZ 2-methyl-4-oxo-3,4 O 2-methyl-4-oxo-3,4- N N H H dihydroquinazolin-8 dihydroquinazolin-8-

ci yl)oxy)ethoxy)ethoxy)ethoxy)ethox yl)oxy)ethoxy)ethoxy)ethoxy)ethox CI CI y)phenyl)acetamide y)phenyl)acetamide 2-((R)-4-(4-chlorophenyl)-2,3,9 2-(R)-4-(4-chlorophenyl)-2,3,9- SS -NN trimethyl-6H-thieno[3,2 trimethyl-6H-thieno[3,2- / NN N 00fl[1,2,4]triazolo[4,3 f][1,2,4]triazolo[4,3- O O O IZ NO O a][1,4]diazepin-6-yl)-N-(4-(2-(2- a] [1,4]diazepin-6-yl)-N-(4-(2-(2 45 N N (2-(2-((3-(2,6-dioxopiperidin-3-yl)- XHH O 02-methyl-4-oxo-3,4 2-methyl-4-oxo-3,4- o CI I N N NH dihydroquinazolin-7 dihydroquinazolin-7- IlT NH N yl)oxy)ethoxy)ethoxy)ethoxy)ethox yl)oxy)ethoxy)ethoxy)ethoxy)ethox. 0 y)phenyl)acetamide y)phenyl)acetamide O

420

Exemplary Estrogen Characterization of Exemplary ReceptorPROTACs Estrogen Receptor 10 Oct 2023

[0951] 5. 5. Table

[0951] Table Characterization PROTACs

Target Ex. Ex. Observed Observed Target ER ER ER ER Engagement DC5o* Dmax* NMR # # [M+H]/Z

[M+H]/Z Engagement IC5o (nM) IC50 (nM) DC50* Dmax* NMR 1 1 743.58 743.58 58.2 58.2 C C B B 610.93 10.93(s, 1H), 10.56-10.43 (s, 1H), 10.56-10.43 (m, (m,1H),1H),9.18-9.13 9.18-9.13(m, (m,1H),1H),7.16-7.13 7.16-7.13(m, (m, 3H), 6.84-6.83(d,(d,J =J 6.4Hz, 3H), 6.84-6.83 = 6.4Hz, 2H),2H),6.69 6.69 (s, 1H),(s, 6.62-6.61 1H), 6.62-6.61 (m, 2H),(m, 2H), 6.55 6.55-

6.52 (m, 6.52 (m, 3H), 6.28-6.26 (d, 3H), 6.28-6.26 (d, JJ == 8.4Hz, 8.4Hz, 2H),2H), 4.99-4.97 4.99-4.97 (m, 1H), 4.29-4.25 (m, 1H), 4.29-4.25 2 2 743.58 743.58 0.79 0.79 B B A (m, 1H), 4.23-4.18 (m, 1H), 4.23-4.18 (m, (m, 1H), 1H), 4.17-4.15 4.17-4.15(m, (m, 1H), 1H),4.06-4.00 4.06-4.00(m, (m,2H), 2H),3.85- 3.85 A 2023248067

3.83 (m, 3.83 (m, 5H), 5H), 3.56-3.53 (m, 1H), 3.34-3.33 (m, 4H), 3.10-3.02 (m, 4H), 3.56-3.53 (m, 1H), 3.34-3.33 (m, 4H), 3.10-3.02 (m, 4H), 3.00-2.85 3.00-2.85 (m, (m, 2H), 2H), 2.60-2.58 2.60-2.58 (m, (m, 3H), 3H), 2.16-2.08 2.16-2.08 (m, (m,1H),1H),1.91-1.88 1.91-1.88(m, (m, 1H), 1.76-1.69 1H), 1.76-1.69 (m, (m, 5H), 5H), 1.43-1.41 1.43-1.41 (m, (m, 2H). 2H). (DMSO-d6, (DMSO-d6,400 400 MHz)MHz) 6:10.96 : 1H), 10.96(s,(s,1H), 9.12 9.12 (s,(s,1H),1H), 7.397.39 (d, J=8.0 (d, J=8.0 1H),- 7.25 Hz, 7.25 Hz, 1H), 6.98 (m,- 6.98 4H),(m, 4H),

6.83 (d, J=6.8 6.83 (d, J=6.8Hz, Hz, 2H),2H), 6.72 6.72 - 6.43 - 6.43 (m, 5H), (m, 5H), 6.26 (d,6.26J=8.6 (d, Hz, J=8.6 2H),Hz, 2H), 5.06 5.06

3 3 743.57 743.57 1.35 1.35 A A (dd, J=5.0, (dd, J=5.0,13.213.2 Hz,Hz, 1H),1H), 4.564.56 - 4.11 - 4.11 (m, 3.94 (m, 3H), 3H), -3.94 3.70 -(m,3.705H),(m, 5H), 3.30 - 3.30 A A 3.25 (m, 3.25 (m, 1H), 3.21 -- 2.77 1H), 3.21 2.77 (m,(m, 8H), 8H), 2.64-2.55 2.64-2.55 (m, 5H), 2.46 - 2.26 (m, (m, 5H), 2.46 - 2.26 (m, 2H), 2H), 2.16 -- 1.94 2.16 1.94 (m, (m, 2H), 1.80 -- 1.22 2H), 1.80 1.22 (m, (m, 7H). 7H). (DMSO-d6, (DMSO-d6, 400 400 MHz) MHz) 610.98 10.98 (s, (s, 1H), 1H), 9.139.13 (s, 1H), (s, 1H), 8.14 8.14 (s, 1H), (s, 1H), 7.40 (d,7.40J= (d, 8.8 J= Hz,8.8 1H),Hz, 1H), 7.17 - 7.17

7.07 (m, 4H), 6.83 - 6.82 (m, 2H), 6.65 - 6.60 (m, 2H), 6.54 - 6.47 (m, 7.07 (m, 4H), 6.83 - 6.82 (m, 2H), 6.65 - 6.60 (m, 2H), 6.54 - 6.47 (m, 3H), 6.27- -6.29 3H), 6.27 6.29(m,(m, 2H), 2H), 5.075.07 (dd, (dd, J= 5.2, J= 5.2, 13.21H), 13.2 Hz, Hz,4.441H),- 4.40 4.44(m,- 4.40 (m, 1H), 4.29 4.29 -4.174.17 (m, (m, 4H), 4H), 3.81 3.81 (t,(t, J= J= 6.4 6.4 Hz, Hz, 2H), 2H), 3.62 3.62 3.60 (m, 3H), 3.53 -- 3.60 4 4 831.65 831.65 1.42 1.42 A A 11H), A A - 3.51 - (m,3H), 3.51 (m, 3H),3.43 3.43 - 3.41 - 3.41 (m,(m, 4H),4H), - 3.17- 3.17 3.24 3.24 (m,2.97 (m, 6H), 6H),- 2.88 - 2.88 (m, 2.97 (m, 4H), 2.78 - 2.74 (m, 3H), 2.61 - 2.56 (m, 2H), 2.44 - 2.37 (m, 2H), 4H), 2.78 - 2.74 (m, 3H), 2.61 - 2.56 (m, 2H), 2.44 - 2.37 (m, 2H), 2.10 - 2.10 1.97 (m, 1.97 (m, 2H), 1.71 -- 1.53 2H), 1.71 1.53 (m,(m, 5H), 5H), 1.411.41 -- 1.38 1.38 (m, (m, 2H). 2H). (DMSO-d6, (DMSO-d6, 400 400 MHz) MHz) 5 5 842.66 842.66 >300 >300 C C 6 6 842.67 842.67 2.18 2.18 C C 7 7 859.68 859.68 102 102 D B B D 610.99 1H), 10.99(s,(s,1H), 9.12 9.12 (s, (s, 1H), 1H), 8.168.16 (s, 1H), (s, 1H), 7.36 7.36 (s, 7.17 (s, 2H), 7.10 -(m, 2H), -7.17 7.10 (m, 3H), 6.83 3H), 6.83(d,(d,J=6.8 J=6.8 Hz,Hz, 2H),2H), 6.66 6.66 - 6.59 - 6.59 (m, 6.52 (m, 2H), 2H),(d,6.52 (d,Hz, J=8.8 J=8.8 2H), Hz, 2H), 8 8 859.68 859.68 0.34 0.34 B B B B 6.50 -- 6.46 6.50 6.46(m, (m,1H), 1H), 6.26 6.26 (d, (d, J=8.4 J=8.4 Hz, 2H), Hz, 2H), 5.10J=5.2, 5.10 (dd, (dd, J=5.2, 13.2 Hz,13.2 1H), Hz, 1H), 4.59 (d, 4.59 (d, J=17.2 J=17.2Hz,Hz, 1H),1H), 4.414.41 (d, J=17.2 (d, J=17.2 1H),- 4.16 Hz,4.22 Hz, 1H), - 4.16 4.22 (m, 3H), (m, 3.823H), 3.82

(t, (t,J=6.0 J=6.0Hz,Hz, 2H), 3.73 -3.67 2H), 3.73 3.67 (m, (m, 2H), 2H),3.60 - 3.55 3.603.55 (m,(m, 2H), 2H), 3.48 3.48 - 3.42(m, - 3.42 (m,

421

3.37- -3.35 2H),3.37 3.35(m,(m, 2H), 3.223.22 2023248067 10 Oct 2023

2H), 2H), (s, 3H), (s, 3H), - 2.82- (m, 3.03 3.03 2.826H), (m,2.82 6H),- 2.69 - 2.69 2.82(m, (m, 10H), 2.63- -2.61 10H), 2.63 2.61(m,(m, 1H), 1H), 2.422.42 - 2.36 - 2.36 (m, 2.15 - 2.15 (m, 1H), 1H), 2.04 -(m, 2.04 1H),(m, 2.03 - 1H), 2.03 1.95 (m, 1.95 1H), (m,1H), 1.81 1.81 - 1.69 - 1.69 (m, (m, 3H),3H), 1.66 1.66 - 1.60- (m, 1.602H), (m,1.58 2H),- 1.50 - 1.50 1.58 (m, (m, 2H). (DMSO-d6,400 2H). MHz) (DMSO-d6, 400 MHz) 9 9 768.61 768.61 >300 >300 D D B B 610.90 1H), 10.90(s,(s,1H), 8.28 8.28 (s, (s, 1H), 1H), 7.197.19 - 7.07 - 7.07 (m, 6.83 (m, 3H), 3H),(d, 6.83 (d, Hz, J=6.4 J=6.4 2H),Hz, 2H),

6.64 (d, 6.64 (d, J=8.4 J=8.4Hz, Hz,1H), 1H), 6.596.59 (s, (s, 2H),2H), 6.52 6.52 (d, J=8.8 (d, J=8.8 Hz,6.49 Hz, 2H), 2H),- 6.44 6.49 - 6.44 (m, 2H),6.19 (m, 2H), 6.19(d,(d,J=8.8 J=8.8 Hz,Hz,2H),2H), - 4.91- 4.91 5.02 5.02 (m, 1H), 1H),(dd, (m,4.96 4.96 (dd,13.2 J=5.2, J=5.2, 13.2 0.86 B A Hz, 1H), 4.26 - 4.19 (m, 1H), 4.14 - 4.06 (m, 2H), 3.82 Hz, 1H), 4.26 - 4.19 (m, 1H), 4.14 - 4.06 (m, 2H), 3.82 (s, 3H), (s, 3H), 3.55 - 3.55 - 3.45 3.45 10 10 768.61 768.61 0.86 B A (m, 2H), 3.30 - 3.10 (m, 12H), 2.98 - 2.85 (m, 3H), 2.59 - 2.53 (m, 1H), (m, 2H), 3.30 - 3.10 (m, 12H), 2.98 - 2.85 (m, 3H), 2.59 - 2.53 (m, 1H),

2.44 -- 2.41 2.44 2.41(m, 1H), (m,1H), 2.38 2.38 - 2.35 - 2.35 (m, (m, 2H), 2H), 2.13 -2.13 2.03- (m, 2.031H),(m,1.951H), 1.95 - 1.87 - 1.87

(m, 1H), 1.75 - 1.65 (m, 3H), 1.48 - 1.33 (m, 3H), 1.26 - 1.12 (m, 2H). (m, 1H), 1.75 - 1.65 (m, 3H), 1.48 - 1.33 (m, 3H), 1.26 - 1.12 (m, 2H).

(DMSO-d6,400 (DMSO-d6, 400 MHz)MHz) 6 10.95 (s, 1H), 9.09 (s, 1H), 7.38 (d, J= 8.03 10.95 (s, 1H), 9.09 (s, 1H), 7.38 (d, J= 8.03 Hz,7.14 Hz, 1H), 1H),(m,7.14 4H),(m,6.834H), 6.83

(d, (d, J= 6.53 Hz, J= 6.53 Hz,2H), 2H), 6.646.64 - 6.59 - 6.59 (m, (m, 2H), 2H), 6.56 - 6.45- (m, 6.56 6.453H),(m,6.19 3H),(d,6.19 J= (d, J=

8.66 Hz, 2H), 5.06 (dd, J= 12.99,5.08 Hz, 1H), 4.36 - 4.27 (m, 1H), 4.19 8.66 Hz, 2H), 5.06 (dd, J= 12.99,5.08 Hz, 1H), 4.36 - 4.27 (m, 1H), 4.19 - 11 11 798.63 798.63 2 2 B B B B 4.18 (m, 4.18 (m,4H), 4H),3.55 3.55 - 3.53 - 3.53 (m, (m, 4H), 4H), 3.26 (s, 3.263H),(s, 3.11 3H),(s,3.114H), (s,2.96 4H), (d,2.96 J= (d, J=

5.9 Hz,2H), 5.9 Hz, 2H),2.69 2.69 - 2.57 (m, 1H), 2.32 - 2.31 (m, 1H), 2.19 (d, J= 6.65 Hz, - 2.57 (m, 1H), 2.32 - 2.31 (m, 1H), 2.19 (d, J= 6.65 Hz,

4H), 2.14 -- 2.04 4H), 2.14 2.04 (m, (m, 3H), 3H), 1.981.98 -- 1.89 1.89 (m,(m, 2H), 11.56 (m, 2H), 11.56 (m, 6H), 6H), 1.29- 1.29- 1.01 1.01 (m, 3H). (DMSO-d6,400 (m, 3H). (DMSO-d6, 400 MHz) MHz) 10.91 (br 10.91 1H),8.20 (brS,s, 1H), 8.20(s,(s,1H),1H), 7.197.19 - 7.09 - 7.09 (m, (m, 3H), 3H), 6.84d,(br 6.84 (br d, J=6.9 J=6.9 Hz, Hz, 2H), 6.67 - 6.58 (m, 5H), 6.54 - 6.43 (m, 2H), 6.30 (d, J=8.5 Hz, 2H), 4.97 2H), 6.67 - 6.58 (m, 5H), 6.54 - 6.43 (m, 2H), 6.30 (d, J=8.5 Hz, 2H), 4.97

12 12 779.56 779.56 11 A A (dd, J=5.1, (dd, J=5.1,13.2 13.2Hz,Hz,1H),1H),4.274.27 - 4.07 - 4.07 (m, 3.83 (m, 5H), 5H), (s, 3.833H),(s,3.39 3H),- 3.28 - 3.28 (m, 3.39 (m, A A 5H), 3.04- -2.85 5H), 3.04 2.85(m,(m,4H),4H),2.592.59 (br S, (br3H),s, 3H), - 2.22- (m, 2.43 2.43 2.224H),(m,2.15 2.15 - 1.88 4H),- 1.88 (m, 4H), (m, 4H), 1.821.82 -- 1.56 1.56 (m, (m, 4H). 4H). (DMSO-d6,400 (DMSO-d6, 400 MHz) MHz) 6 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.07 (m,6.82 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.07 (m, 3H), 3H),(d,6.82 (d, J=6.8Hz, J=6.8 Hz,2H), 2H), 6.696.69 - 6.58- 6.58 (m, (m, 3H), 3H), 6.55 -6.556.47- (m, 6.47 (m,6.26 4H), 4H), (d,6.26 J=8.8(d,Hz,J=8.8 Hz, 2H), 4.96 (dd, J=5.2, 13.2 Hz, 1H), 4.29 - 4.06 (m, 3H), 3.85 - 3.78 (m, 13 13 743.58 743.58 0.37 0.37 A A 2H), 4.96 (dd, J=5.2, 13.2 Hz, 1H), 4.29 - 4.06 (m, 3H), 3.85 - 3.78 (m, A A 5H), 3.30- -3.28 5H), 3.30 3.28(m,(m, 4H), 4H), 3.043.04- 2.80- 2.80 (m, 2.60 (m, 3H), 2.52 -(m, 3H), -2.60 2.526H), (m, 6H), 2.45 - 2.45

2.34(m,3H), 2.34(m, 3H), 2.17 - 1.99 (m, 1H), 1.97 - 1.82 (m, 1H), 1.76 - 1.58 (m, 3H), 2.17 - 1.99 (m, 1H), 1.97 - 1.82 (m, 1H), 1.76 - 1.58 (m, 3H),

1.56 -- 1.45 1.56 1.45 (m, (m, 2H), 2H), 1.431.43 -- 1.291.29(m, (m,2H). 2H).(DMSO-d6, (DMSO-d6, 400 400 MHz) MHz) 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.08 (m, 3H), 6.82 (d, 14 14 743.58 743.58 0.49 0.49 A A 6 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.08 (m, 3H), 6.82 (d, A A J=6.8Hz, J=6.8 Hz,2H), 2H), 6.666.66 - 6.58- 6.58 (m, (m, 3H), 3H), 6.55 -6.556.47- (m, 6.47 (m,6.26 4H), 4H), (d,6.26 J=8.8(d,Hz,J=8.8 Hz,

422

2H), 4.96 4.96 (dd, (dd, J=5.2, Hz, 1H), J=5.2, 13.2 Hz, 1H), 4.28 -4.10 4.10(m, (m,3H), 3.85- -3.78 3H),3.85 3.78(m, (m, 2023248067 10 Oct 2023

2H), 5H), 3.30- -3.28 5H), 3.30 3.28(m,(m, 4H), 4H), 3.053.05 - 2.80 - 2.80 (m, 2.58 (m, 3H), 2.51 -(m, 3H),-2.58 2.51 (m, 6H), 6H), 2.38 - 2.38 2.33 (m, 2.33 (m,2H), 2H),2.32 2.32 - 2.24 - 2.24 (m, (m, 1H), 2.18 2.18 1H), - 2.00- (m, 2.001H), (m,1.97 - 1.86 - 1.97(m, 1H), 1.86 (m, 1H), 1.74 1H), 1.74- -1.581.58(m,(m, 3H),3H), 1.551.55 - 1.44- 1.44 (m, 2H), (m, 2H), 1.34 -(m, 1.43 - 1.43 1.34 2H).(m, 2H). (DMSO-d6,400 (DMSO-d6, 400 MHz)MHz) 6 10.91 (s, 1H), 8.23 (s, 2H), 7.17 - 7.09 3H), 10.91 (s, 1H), 8.23 (s, 2H), 7.17 - 7.09 (m, (m, 6.83 3H),(d, 6.83 J=6.8 (d, Hz, J=6.82H), Hz, 2H), 6.64 (d, 6.64 (d, J=8.4 J=8.4Hz, Hz, 1H), 1H), 6.596.59 (s, 2H), (s, 2H), 6.53 6.53 (d, J=8.8 (d, J=8.8 Hz, 2H), Hz,6.492H), 6.49 - 6.45 - 6.45

(m, 2H), (m, 2H),6.20 6.20(d,(d,J=8.8 J=8.8 Hz, Hz,2H),2H), 4.96 4.96 (dd, J=5.0, (dd, J=5.0, 13.2 Hz, 13.2 1H), Hz, 1H), 4.25 - 4.194.25 - 4.19 15 15 754.60 754.60 1.7 1.7 A A (m, 1H), 4.14 (m, 1H), 4.14 -- 4.06 4.06 (m, (m, 1H), 1H), 4.15 4.15 - - 4.06 4.06 (m, (m, 1H), 1H), 3.843.84 -3.80 3.80 (m,3H), (m, 3H), V V 3.51 (d, J=9.2 Hz, 7H), 3.28 (s, 4H), 2.98 - 2.83 (m, 1H), 3.03 (m, 3.51 (d, J=9.2 Hz, 7H), 3.28 (s, 4H), 2.98 - 2.83 (m, 1H), 3.03 - 2.82 - 2.82 (m, 2H),2.58 2H), 2.58(s, 1H),2.32 (s,1H), 2.32 - 2.26 - 2.26 1H),1H), (m, (m, - 2.04- (m, 2.22 2.22 2.044H),(m,1.944H),- 1.87 1.94(m,- 1.87 (m, 1H), 1.80 1H), 1.80 -- 1.55 1.55 (m, (m, 5H), 1.21 -- 1.11 5H), 1.21 1.11 (m,(m, 2H). 2H). (DMSO-d6,400 (DMSO-d6, 400 MHz) MHz) 610.90 1H), 10.90(s,(s,1H), 9.13 9.13 (s, (s, 1H),1H), 8.148.14 (s, 1H), (s, 1H), - 6.92- (m, 7.01 7.01 6.922H), (m,6.87 2H), 6.87 - 6.79 - 6.79

(m, 2H), (m, 2H),6.65 6.65(d,(d,J=8.4 J=8.4 Hz,Hz, 1H),1H), 6.60 6.60 (s, 2H),(s, 6.55 2H), (d,6.55 (d, Hz, J=8.8 J=8.8 2H),Hz,6.512H), 6.51

- 6.44 (m, 2H), 6.26 (d, J=8.4 Hz, 2H), 4.96 (dd, J=5.4, 13.4 Hz, 1H), 4.75 - 6.44 (m, 2H), 6.26 (d, J=8.4 Hz, 2H), 4.96 (dd, J=5.4, 13.4 Hz, 1H), 4.75 16 91 799.6 9'66L 0.82 0.82 A B V (t, J=4.8 (t, Hz,1H), J=4.8 Hz, 1H),4.27 4.27 - 4.06 (m, 3H), 3.85 - 3.80 (m,3.29 - 4.06 (m, 3H), 3.85 - 3.80 (m, 3H), 3H),- 3.25 3.29 (m,- 3.25 (m, 6H), 2.99 -- 2.84 6H), 2.99 2.84 (m,(m, 3H), 2.54 (d, (d, J=4.4 J=4.4 Hz, Hz, 8H), 2.19 -1.86 8H), 2.19 1.86(m,(m,4H),4H),1.85 1.85 - 1.58 - 1.58 (m, 5H), 1.24 -- 1.07 (m, 5H), 1.07 (m,(m, 2H). 2H). (DMSO-d6, (DMSO-d6, 400 400 MHz)MHz) 610.91 1H), 10.91(s,(s,1H), 9.149.14(s, (s, 1H),1H), 7.857.85 (br S, s, 1H), (br1H), 7.48 - 7.34- (m, 7.48 7.342H),(m,7.18 2H), - 7.18

7.10 (m, 7.10 (m,3H),3H),6.836.83(br(br d, J=6.7 d, J=6.7 Hz, 2H), Hz, 2H), 6.69 -6.696.60 -(m, 6.60 5H),(m, 5H), 6.54 6.54 - 6.47 - 6.47

(m, 2H), 6.30 (d, J=8.5 Hz, 2H), 5.08 - 4.94 (m, 3H), 4.28 - 4.17 (m, 2H), (m, 2H), 6.30 (d, J=8.5 Hz, 2H), 5.08 - 4.94 (m, 3H), 4.28 - 4.17 (m, 2H),

17 LI 778.57 778.57 1.5 1.5 B B 4.16 -- 4.08 4.16 4.08(m, (m,1H),1H),3.843.84(s, (s, 4H), 4H), 3.663.66(br S, s, 1H), (br1H), 3.04 - 2.83- (m, 3.04 2.834H), (m,2.824H), 2.82 - 2.71 (m, 1H), 2.68 (br s, 1H), 2.63 - 2.54 (m, 2H), 2.48 - 2.26 (m, 2H), - 2.71 (m, 1H), 2.68 (br S, 1H), 2.63 - 2.54 (m, 2H), 2.48 - 2.26 (m, 2H),

2.15 -- 2.03 2.15 2.03(m, (m,1H),1H), 1.97 1.97 - 1.88 - 1.88 (m, 1H), (m, 1H), 1.71 (br1.71d,(br J=7.5d, J=7.5 Hz, 1H). Hz, 1H). (DMSO-d6,400 (DMSO-d6, 400 MHz)MHz) 18 18 790.59 790.59 17.5 17.5 B B 610.89 1H), 10.89(s,(s,1H), 8.19 8.19 (s, (s, 1H), 1H), 7.227.22 - 7.16 - 7.16 (m, 6.90 (m, 3H), 3H),(br d, J 6.90 = 6.8 (br Hz, d, J= 6.8 Hz, 2H),6.68 2H), 6.68(d, (d,J J= 8.4Hz,Hz, = 8.4 1H), 1H), 6.616.61 (brJ d, (br d, J= Hz, = 9.2 9.2 2H), Hz, 6.54 2H), 6.54 - 6.50 (m, - 6.50 (m, 1H), 6.47 (s, 1H), 5.87 (d, J = 11.2 Hz, 2H), 4.95 (dd, J = 5.2, 13.2 Hz, 19 61 790.58 790.58 4.5 4.5 B A 1H), 6.47 (s, 1H), 5.87 (d, J= 11.2 Hz, 2H), 4.95 (dd, J= 5.2,13.2 Hz, V 1H), 4.25- -4.20 1H), 4.25 4.20(m,(m, 2H), 2H), 4.134.13 - 4.07 - 4.07 (m, 1H), 1H), (m, 3.82 (s, 3.82 (s,3.27 3H), 3H),- 3.25 - 3.25 3.27 (m, (m, 6H), 3.03 - 2.83 (m, 9H), 2.19 (br d, J= 7.2 3H), 6H), 3.03 - 2.83 (m, 9H), 2.19 (br d, J = 7.2 Hz, Hz, 2.07 1.89 - 3H),- 2.07 (m, 3H),(m, 3H), 1.89 1.76 -- 1.58 1.76 1.58 (m, (m, 5H), 5H), 1.16 1.16 (br (brd,d,J=J =9.2 Hz,Hz,2H). 9.2 2H).(DMSO-d6,400 (DMSO-d6, 400 MHz)MHz)

423

610.96 10.96(s, 1H), 8.20 (s, 1H), (s, 1H), 8.20 (s, 7.62 -7.20 1H), 7.62 7.20(m, 1H),7.18 (m,1H), - 7.05 7.187.05 (m,(m, 3H), 6.94 2023248067 10 Oct 2023

3H), 6.94 (s, 1H), 6.82 (d, J=6.4 Hz, 2H), 6.71 (s, 1H), 6.67 - 6.58 (m, 2H), 6.56 (s, 1H), 6.82 (d, J=6.4 Hz, 2H), 6.71 (s, 1H), 6.67 - 6.58 (m, 2H), 6.56 -

6.43 (m, 6.43 (m,3H), 6.26 3H), 6.26 (d, (d, J=8.8 J=8.8 Hz, Hz, 2H), 2H), 5.00J=5.2, 5.00 (dd, (dd, J=5.2,13.2 13.2 Hz, 1H),Hz, 4.371H), - 4.37 20 20 779.6 779.6 1.2 1.2 B B 4.29(m, 4.29 4.26 1H), 4.26 (i,1H), - 4.14 - 4.14 (m, (m, 2H), 2H), 3.81 (t, (t, J=6.4 3.81J=6.4 Hz, 2H), Hz,3.31 2H),- 3.27 - 3.27 3.31 (m, (m, 5H), 3.04- -2.82 5H), 3.04 2.82(m,(m, 3H),3H), 2.642.64 - 2.52- 2.52 (m, 2H),(m, 2.48 2H),-2.42 2.48 (m,-2.423H),(m,2.41 3H), - 2.41 2.25 (m, 2.25 (m,3H), 3H),2.172.17 - 2.02 - 2.02 1H), 1H), (m, (m, 2.00 - 2.001.90- (m, 1.901H), (m,1.751H), - 1.59 1.75(m, - 1.59 (m, 3H), 1.53 -- 1.43 (m, 3H), 1.53 (m, 2H), 2H), 1.421.42 -- 1.32 1.32 (m, (m, 2H). 2H). (DMSO-d6,400 (DMSO-d6, 400 MHz) MHz) 6 10.98 (s, 1H), 8.15 (s, 1H), 7.68 - 7.34 (m, 1H), 7.21 - 7.05 (m, 3H), 6.95 10.98 (s, 1H), 8.15 (s, 1H), 7.68 - 7.34 (m, 1H), 7.21 - 7.05 (m, 3H), 6.95

(s, 1H), (s, 6.84(d, 1H), 6.84 (d, J=7.2 J=7.2Hz,Hz, 2H), 2H), 6.736.73 (s, 1H), (s, 1H), - 6.58- (m, 6.68 6.68 6.584H), (m,6.50 4H),(d,6.50 (d, 1H), 4.42 21 21 815.6 9'9'8 2.5 2.5 B B J=8.2 Hz, 1H), 6.31 (d, J=8.4 Hz, 2H), 5.02 (dd, J=4.8, 13.2 Hz,1H), J=8.2 Hz, 1H), 6.31 (d, J=8.4 Hz, 2H), 5.02 (dd, J=4.8, 13.2 Hz, 4.42 -- 4.27 4.27 (m, 1H),4.27 (m,1H), 4.27 - 4.04 - 4.04 (m, (m, 4H),4H), 3.31 3.31 (s, 4H),(s, 4H), 2.78 -(m, 3.03 -3.03 2.783H),(,3H), 2.68 2.68 -- 2.55 2.55 (m, 1H),2.48 (m,1H), 2.48 (s,(s, 6H), 6H), 2.41 2.41 - 2.32 - 2.32 (m, 3H), (m, 3H), 1.90 -(m, 2.16 -2.16 1.904H), (,4H), 1.72 1.72 (m,1H), (m, 1H), 1.63 1.63 (m,(m, 2H). 2H). (DMSO-d6,400 (DMSO-d6, 400 MHz) MHz) 6 10.97 (s, 1H), 8.19 (s, 1H), 7.68 - 7.20 (m, 1H), 7.18 (m, 10.97 (s, 1H), 8.19 (s, 1H), 7.68 - 7.20 (m, 1H), 7.18 - 7.07 - 7.07 3H),(,3H), 6.94 6.94 (s, 1H), (s, 6.83(d, 1H), 6.83 (d, J=6.4 J=6.4Hz,Hz, 2H),2H), 6.716.71 (s, 1H),(s, 1H), - 6.57- (m, 6.66 6.66 6.572H),(m,6.55 2H), - 6.55

6.43 (m, 6.43 (m,3H), 3H),6.19 6.19(d, (d, J=8.4 J=8.4 Hz, Hz, 2H), 2H), 5.00 (dd,5.00J=5.0, (dd, J=5.0,13.2 13.2 Hz, 1H),Hz, 4.38 1H), - 4.38 22 zz 804.6 804.6 4.4 4.4 B A 4.28 (m, 4.28 1H), 4.26 (m, 1H), 4.26 4.17- 4.17(m,(m,1H), 1H),4.12 4.12 (d,(d,J=4.6 J=4.6Hz, 1H),3.30 Hz,1H), 3.30(s, (s, 9H), 9H), V 3.01 -- 2.78 3.01 2.78(m, (m,4H), 4H), 2.712.71 - 2.55 - 2.55 (m, (m, 2H), 2H), 2.44 -2.442.26- (m, 2.26 (m,2.16 5H), 5H), 2.16 - 2.03 - 2.03

(i,1H), (m, 1H), 2.02 - 1.89 2.02- 1.89 (m,(m,1H),1H), 1.79 1.79 - 1.62 - 1.62 (m, 3H),(m,1.40 3H),(m,1.403H),(m, 1.27 3H), - 1.27 1.06 (m, 1.06 (m,2H).2H).(DMSO-d6,400 (DMSO-d6, 400 MHz) MHz) 610.96 10.96(s, 1H), 9.12 (s, 1H), 9.12 (s,(s, 1H), 1H), 8.16 8.16 (s,(s,1H), 1H),7.637.63- 7.20 7.20 (m, 1H), 7.18 (m, 1H), 7.18 -- 7.06 (m, 3H), (m, 3H),6.946.94 (s,(s, 1H), 1H), 6.816.81 (d, (d, J=6.4 J=6.4 Hz, 2H), Hz, 2H), 6.71 (s,6.71 (s, 1H), 6.67 (m, 1H), 6.67 - 6.58 - 6.58 (m, 2H), 6.51 - 6.37 (,3H), 6.24 (d, J=8.4 Hz, 2H),(dd, 2H), 6.51 - 6.37 (m, 3H), 6.24 (d, J=8.4 Hz, 2H), 5.00 5.00 (dd, 13.2 J=5.2, J=5.2, Hz, 13.2 Hz, 23 er 791.6 791.6 1.8 8'I B B 8 1H), 4.71 1H), 4.71- -4.60 4.60(m,(i,1H), 1H), 4.404.40 - 4.29 - 4.29 (m, 1H), (m, 1H), 4.13 -(m, 4.26 -4.26 4.132H),(m, 2H), 3.32 - 3.32

3.27 (m, 3.27 (m,9H), 9H),3.043.04 - 2.80 - 2.80 (m, (m, 3H),3H), - 2.54- (m, 2.63 2.63 2.542H), (m,2.422H), 2.42(m, - 2.31 - 2.31 (m, 1H), 2.30 1H), 2.30- -2.19 2.19(m,(m, 3H),3H), 2.132.13 - 2.03 - 2.03 (m, 2.02 (m, 4H), 4H),-2.021.91 -(m,1.911H),(m,1.74 1H), - 1.74 1.51 (m, 3H). (DMSO-d6,400 1.51 (m, 3H). (DMSO-d6, 400 MHz) MHz) 610.89 10.89(s,(s,1H),1H),9.179.17(s, (s, 1H),1H), 8.328.32 (s, 1H), (s, 1H), - 7.10- (m, 7.21 7.21 7.103H),(m,6.863H), (d,6.86 (d, J=6.4 Hz, 2H), 6.68 - 6.58 (m, 4H), 6.52 - 6.44 (m, 2H), 6.08 (d, J=8.0 Hz, J=6.4 Hz, 2H), 6.68 - 6.58 (m, 4H), 6.52 - 6.44 (m, 2H), 6.08 (d, J=8.0 Hz,

0.7 B A 1H), 5.97 (d, J=14.2 Hz, 1H), 4.95 (dd, 13.2 Hz, 1H),Hz, 1H), 5.97 (d, J=14.2 Hz, 1H), 4.95 (dd, J=5.2,J=5.2,13.2 1H), 4.28 4.28 - 4.16 - 4.16 24 24 786.6 786.6 0.7 A (m, 2H), (m, 2H),4.14 - 4.05 (m, 1H), 3.83 (s, 3H), 3.18 (s, 2H), 2.99 (m, 4.14- 4.05 (m, 1H), 3.83 (s, 3H), 3.18 (s, 2H), 2.99 - 2.85 - 2.85 (m, 3H), 2.54 - 2.52(m,(m, 3H), 2.54 - 2.52 13H), 13H), 2.122.12 - 1.86 - 1.86 (m, 3H), (m, 1.71 3H),(d,1.71J=10.8 (d, Hz, J=10.8 3H), Hz, 3H), 1.48 -- 1.19 1.48 1.19 (m,(m, 6H). 6H). (DMSO-d6,400 (DMSO-d6, 400 MHz) MHz)

424

25 715.6 0.5 C 2023248067 10 Oct 2023

25 715.6 0.5 C 610.90 1H), 10.90(s,(s,1H), 9.16 (s, (s, 9.16 1H), 1H), 7.197.19 - 7.09 - 7.09 (m, 6.82 (m, 3H), 3H),(br 6.82 (br d, J = 6.6 Hz,6.6 Hz, d, J= 6.68- -6.57 2H), 6.68 2H), 6.57(m,(m, 3H), 3H), 6.536.53 - 6.40 - 6.40 (m, 6.29 (m, 4H), 6.23 -(m, 4H),- 6.29 6.23 2H),(m, 2H), 4.96 4.96 (dd, JJ= (dd, 5.1, 13.3 = 5.1, 13.3Hz, 1H), Hz,1H), 4.534.53 (quin, (quin, J= Hz, J = 7.3 7.3 1H), 1H),-4.31 Hz, 4.31 4.01 -(m, 4.013H),(m, 3H),

26 26 769.6 769.6 0.7 0.7 A A 3.83 (s, 3.83 (s, 3H), 3H), 3.633.63(br(brS,s,1H), 1H),3.333.33 - 3.23 - 3.23 (m, (m, 11H),11H), 3.10 J=8.8, 3.10 (td, (td, J=8.8, 17.4 17.4 A A Hz, 1H), 3.03 - 2.79 (m, 3H), 2.55 (br s, 2H), 2.46 - 2.25 (m, 2H), 2.16 Hz, 1H), 3.03 - 2.79 (m, 3H), 2.55 (br S, 2H), 2.46 - 2.25 (m, 2H), 2.16 -

2.00 (m, 2.00 (m, 3H), 3H), 1.99 1.87 (m, 1H), 1.70 1.99 -- 1.87 1.70 (br(br d,d,J=6.0 J=6.0 Hz, Hz,1H). 1H). (DMSO-d6, (DMSO-d6, 400 MHz) 400 MHz) 610.96 1H), 10.96(s,(s,1H), 9.13 9.13 (br (br s, 1H), S, 1H), 8.208.20 - 7.39 - 7.39 1H),- 7.22 (m, 7.22 (m, 1H), - 7.08 7.08 (m, 3H),(m, 3H),

6.97 - 6.93 (m, 1H), 6.81 (br d, J=7.7 Hz, 2H), 6.73 - 6.69 (m, 1H), 6.97 - 6.93 (m, 1H), 6.81 (br d, J=7.7 Hz, 2H), 6.73 - 6.69 (m, 1H), 6.67 - 6.67

6.62 (m, 6.62 (m,1H),1H),6.606.60(d,(d, J=2.3 J=2.3 1H), 1H), Hz, Hz, 6.50 -6.506.38 - (m, 6.383H),(m,6.273H), 6.27(m, - 6.21 - 6.21 (m, 27 27 807.6 807.6 0.5 0.5 A B B 2H), 5.00 (br dd, J=5.1, 13.2 Hz, 1H), 4.65 (br t, J=5.8 Hz,4.37 2H), 5.00 (br dd, J=5.1, 13.2 Hz, 1H), 4.65 (br t, J=5.8 Hz, 1H), 1H), - 4.37 A 4.30 (m, 4.30 1H),4.25 (m,1H), 4.25 - 4.18 - 4.18 1H), 1H), (m, (m, 4.16 4.16 (br d,(br d, J=4.9 J=4.9 Hz, 1H), Hz, 1H), 3.28 3.28 - 3.25 - 3.25

(m, 6H), 3.02 - 2.81 (m, 3H), 2.61 - 2.53 (m, 4H), 2.45 - 2.37 (m, 4H), (m, 6H), 3.02 - 2.81 (m, 3H), 2.61 - 2.53 (m, 4H), 2.45 - 2.37 (m, 4H),

2.12 -- 1.90 2.12 1.90 (m, (m, 5H), 1.76 -- 1.64 5H), 1.76 1.64 (m,(m, 3H). 3H). (DMSO-d6, (DMSO-d6, 400 400MHz)MHz) 6 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.08 (m, 3H), 6.81 10.91 (s, 1H), 9.13 (s, 1H), 8.14 (s, 1H), 7.18 - 7.08 (m, 3H), 6.81 (br d, (br d,

J=7.7Hz, J=7.7 Hz,2H), 2H), 6.666.66 - 6.59 - 6.59 (m, (m, 3H), 3H), 6.39 -(m, 6.51 -6.51 6.394H), (m,6.27 4H), 6.27(m,- 6.22 (m, - 6.22

2H),4.96 2H), 4.96(br(brdd,dd,J=5.1, J=5.1, 12.912.9 Hz, Hz, 1H), 1H), 4.65 4.65 (br t,(brJ=6.3t, J=6.3 Hz,4.26 Hz, 1H), 1H),- 4.26

28 28 771.6 771.6 0.3 0.3 A A 4.19 (m, 4.19 1H),4.16 (m,1H), 4.16 (br(br d, J=4.6 d, J=4.6 Hz, 1H), Hz, 1H), 4.07 -(m, 4.13 -4.13 4.07 1H),(m, 1H), 3.84 3.84 - 3.80 - 3.80 A A (m, 3H), (m, 3H),3.29 - 3.23 3.29- 3.23 (m,(m,5H),5H), 3.00 3.00 - 2.84 - 2.84 (m, 3H), 2.62 - 2.52 (m, 8H), (m, 3H), 2.62 - 2.52 (m, 8H),

2.35 2.35 (br(br s,S,2H), 2H),2.09 2.09- -1.881.88(m, (m,4H), 4H),1.781.78- -1.661.66 (m, (m,3H). 3H).(DMSO-d6, (DMSO-d6, 400 400 MHz) MHz) 29 29 750.6 750.6 2.4 2.4 C C 610.87 10.87 (s,(s, 1H),1H),9.209.20 (s, 1H), (s, 1H), 8.26 8.26 (s, 1H), (s, 1H), 7.46J=8.8 7.46 (d, (d, J=8.8 Hz, 1H),Hz, 1H), 7.24 - 7.24 7.07 (m,3H), 7.07 (m, 3H),6.996.99 (d,(d, J=8.8 J=8.8 1H), 1H), Hz, Hz, 6.86J=6.8 6.86 (d, (d, J=6.8 Hz, 2H), Hz, 2H), 6.75 6.75 - 6.56 - 6.56

(m, 4H), (m, 4H),6.50 6.50(d,(d,J=8.0 J=8.0Hz,Hz, 1H),1H), 6.09 6.09 (d, J=8.0 (d, J=8.0 Hz,5.97 Hz, 1H), 1H),(d,5.97 (d, J=14.4 J=14.4

Hz, 1H), 4.18 (d, J=4.4 Hz, 1H), 3.89 (s, 1H), 3.35 - 3.23 (m, 8H), 3.19 (d, 3.19 (d, 30 30 768.6 768.6 2.1 2.1 B B Hz, 1H), 4.18 (d, J=4.4 Hz, 1H), 3.89 (s, 1H), 3.35 - 3.23 (m, 8H), B B J=6.8Hz, J=6.8 Hz,3H), 3H), 3.053.05 - 2.84 - 2.84 (m, 2H), (m, 2H), 2.60- (m, 2.76 -2.76 2.602H), (m,2.54 2H),(s,2.54 3H), (s, 2.203H), 2.20 (d, J=6.8 Hz, 2H), 2.06 (dd, J=6.0,12.0 Hz,1H), 1.83 (s, 1H), 1.75 (d, (d, J=6.8 Hz, 2H), 2.06 (dd, J=6.0, 12.0 Hz, 1H), 1.83 (s, 1H), 1.75 (d,

J=12.0Hz, J=12.0 Hz, 3H), 3H), 1.621.62 1H),1H), (s, (s, 1.54 1.54 - 1.44- (m, 1.442H), (m,1.44 2H),- 1.29 - 1.29 1.44 (m, 2H), (m, 2H),

1.21 (d, J=10.0 Hz, 2H). (DMSO-d6,400 MHz) 1.21 (d, J=10.0 Hz, 2H). (DMSO-d6, 400 MHz) 31 31 C C

[0952]

[0952] DC 5 o(nM) *ER DC50 *ER 1<=B<10; A<1;1<=B<10; (nM)A<1; 10<=C<100; 10<=C<100; D>=100 D>=100

425

[09531

[0953] **ER Dmax(%) A>=75; **ER Dmax(%) 50<=B<75;C<50 A>=75;50<=B<75; C<50

[0954]

[0954] Table 6. Characterization Table 6. of Exemplary Characterization of Androgen Exemplary Androgen Receptor Receptor PROTACs PROTACs

Ex. # Ex. # m/z observed m/z observed AR * DC AR NMR DC5o* DC50* Dmax** Dmax** NMR 32 32 824.54 824.54 A A 33 33 852.58 852.58 A A 1HNMR 1H NMR (400 (400 MHz, MHz, d6-DMSO): d6-DMSO): 10.88 . (s, 10.88 1H),(s,8.22 1H),(s, 8.22 (s, 7.91 1H), 1H), (d, 7.91J=8.8 (d, J=8.8 Hz, 1H), 7.74 Hz, 1H), 7.74 (d, (d, J=8.8 J=8.8 Hz, Hz, 2H), 7.53 -- 7.45 2H), 7.53 7.45 (m, (m, 2H), 2H), 7.21 7.21 (d, (d,J=2.4 Hz, 1H), J=2.4 Hz, 1H), 6.99 (dd, J=9.2, 17.6 Hz, 4H), 6.73 (s, 1H), 4.33 (s, 4.331H), 4.06 4.06 (d, (s, 1H), (d, Hz, J=9.2 1H),Hz, 1H), J=9.2 34 34 832.61 832.61 C 6.99 (dd, J=9.2, 17.6 Hz, 4H), 6.73 (s, 1H), C 3.86 (d, 3.86 (d, J=12.4 J=12.4Hz,Hz,3H),3H), 3.323.32 - 3.29 - 3.29 (m, 2.80 (m, 9H), 9H),(t, 2.80 (t, J=12.0 J=12.0 Hz, 3H),Hz, 2.593H), - 2.59 2.54 (m, 2.54 (m, 4H), 2.22 (d, 4H), 2.22 (d, J=6.8 J=6.8 Hz, 2H), 1.81 (d, 2H), 1.81 (d, J=10.3 Hz, 4H), J=10.3 Hz, 4H), 1.55 1.55 -- 1.47 1.47 (m, (m, 2H), 1.45 2H), 1.45- -1.31 1.31(m,(m, 2H), 2H), 1.251.25 - 1.17 - 1.17 (s, 8H), (s, 8H), 1.136H) 1.13 (s, (s, 6H) 35 35 849.6 849.6 C C 36 36 849.61 849.61 C C 37 37 877.64 877.64 C C 38 38 877.64 877.64 C C H NMR ¹H NMR (400 (400 MHz, MHz, DMSO-d) DMSO-d) 1.12 (6H, s), (6H, 6 1.12 1.21 s), (6H, s),(6H, 1.21 1.43-1.54 (4H, m), s), 1.43-1.54 (4H, m), 810.3 810.3 1.74-1.78 (2H, m), 1.88-1.91 (1H, m), 2.30-2.44 1.74-1.78 (2H, m), 1.88-1.91 (1H, m), 2.30-2.44 (8H, (8H, m), m),2.90-2.97 2.90-2.97(3H, m), (3H,m), 46 46 A A 3.42-3.59 (7H, 3.42-3.59 (7H, m), m), 4.03-4.07 4.03-4.07 (3H, (3H, m), m), 4.30 (1H,s), 4.30(1H, s), 6.86-6.91 6.86-6.91 (3H, (3H, m), m), 6.99- 6.99 A A 7.02 (1H,m), 7.02 (1H, m),7.22 7.22(1H,(1H, d, Jd,= J= 2.4 2.4 Hz), Hz), 7.64 7.64 (1H, d, (1H, J = d, 8.8J= Hz),8.87.79 Hz), 7.79 (1H, d, J(1H, = d, J= 8.8 Hz), 7.90-7.97 (2H, m), 8.62 (1H, d, J= 2.0 Hz), 10.90 (1H, s). 8.8 Hz), 7.90-7.97 (2H, m), 8.62 (1H, d, J = 2.0 Hz), 10.90 (1H, s). ¹HH NMR NMR (400 (400 MHz, MHz, DMSO-d DMSO-d) 1.12 6(6H, s), (6H, ) 6 1.12 1.21 s),(6H, s),(6H, 1.21 1.37-1.58 (4H, m), s), 1.37-1.58 (4H, m), 1.73-1.81 1.73-1.81 (2H, m), m), 1.86-1.91 1.86-1.91 (1H,(1H, m),m), 2.30-2.37 2.30-2.37 (2H, (2H,m), m),2.40-2.46 2.40-2.46(2H, (2H,m),m), 47 47 824.3 824.3 B B 2.82-2.91(1H, m), 3.30-3.35 (4H, m), 3.55-3.65 (4H, m), 4.03-4.30 (6H, m), 2.82-2.91 (1H, m), 3.30-3.35 (4H, m), 3.55-3.65 (4H, m), 4.03-4.30 (6H, m), B B 5.54-5.63(1H, 5.54-5.63 (1H,m),m),6.876.87 (1H,(1H, d, J d, J=Hz), = 9.6 9.6 6.96-7.07 Hz), 6.96-7.07 (3H, m),(3H,7.21 m),(1H,7.21 d, J (1H, = d, J= 2.4 Hz), 2.4 Hz),7.63 7.63(1H,(1H,d, d, J =J=9.69.6 Hz),Hz), 7.90-8.04 7.90-8.04 (3H, m),(3H,8.62 m), 8.62 (1H, d, J= 2.4 Hz), (1H, d, J = 2.4 Hz),

10.93(1H, 10.93 (1H,s).s). ¹HH NMR NMR (400 (400 MHz, MHz, DMSO-d) DMSO-d) 1.12 (6H,1.12 s), (6H, 1.21 s),(6H, s),(6H, 1.21 1.43-1.47 (2H, m), s), 1.43-1.47 (2H, m), 1.49-1.53 (2H, m), 1.73-1.78 (2H, m), 2.13-2.17 (1H, m),(1H, (2H, 2.32 2.32 m), = 7.2 t, J= 7.2 t, J (2H, 798.6 798.6 1.49-1.53 (2H, m), 1.73-1.78 (2H, m), 2.13-2.17 48 48 A B B Hz), 2.43-2.47 Hz), 2.43-2.47 (5H, (5H, m), m), 2.61-2.62 2.61-2.62 (1H,(1H, m), m), 2.87-2.93 2.87-2.93 (1H, (1H,m), m),3.59 3.59(4H, (4H,s), s), A 4.01-4.07 (3H, m), 4.30 (1H, s), 5.28 (1H, dd, J= 12.4, 5.2 Hz), 6.35 (1H, dd, J 4.01-4.07 (3H, m), 4.30 (1H, s), 5.28 (1H, dd, J = 12.4, 5.2 Hz), 6.35 (1H, dd, J = 8.0, 2.4 = 8.0, 2.4 Hz), Hz),6.526.52(1H, (1H, d, Jd,=1.6 J=1.6Hz),Hz), 6.86 6.86 (1H, d,(1H,J = d, J= 8.8 8.87.00 Hz), Hz),(1H,7.00 dd, (1H, J dd, J

426

8.8, 2.4 2.4 Hz), Hz),7.21 7.21(1H, (1H, d, =J= 2.4 2.4 Hz),Hz), 7.63 7.63 d, J =d,9.2 (1H, (1H, J=Hz), 9.27.80 Hz),(1H, 7.80 d, (1H, d, J 2023248067 10 Oct 2023

==8.8, d, J J

8.0 Hz), == 8.0 Hz),7.90 (1H, 7.90(1H, d, Jd,J= = 8.8 8.8 Hz),Hz), 7.95 7.95 (1H, (1H, dd, J dd,J= = 9.2, 2.4 9.2,2.4 Hz), 8.62 Hz), (1H,8.62d, J(1H, d,J 2.4 Hz), == 2.4 Hz),11.0911.09 (1H, (1H, s). s). ¹HH NMRNMR (400(400 MHz,MHz, DMSO-d) DMSO-d) 1.12 (6H, s), (6H, 81.12 1.21 s),(6H, s),(6H, 1.21 1.44-1.48 (2H, m), s), 1.44-1.48 (2H, m), 1.52-1.58(2H, 1.52-1.58 (2H,m),m), 1.74-1.79 1.74-1.79 (2H, (2H, m), 2.15-2.19 m), 2.15-2.19 (1H, m),(1H,2.30 m),(2H, 2.30 t, J (2H, = 7.2 t, J= 7.2

Hz), Hz), 2.43-2.50 2.43-2.50 (4H, (4H, m),m), 2.51-2.67 2.51-2.67 (2H, (2H, m), m), 2.86-2.95 2.86-2.95 (1H, (1H,m), m),3.60 3.60(4H, (4H,s), s), 3.97 3.97 798.6 A A (2H, t, J = 6.4 Hz), 4.05 (1H, d, J = 9.2 Hz),Hz), (2H, t, J= 6.4 Hz), 4.05 (1H, d, J= 9.2 (1H, (1H, 4.30 4.30 s), 5.38 s), 5.38 (1H, (1H, dd, J =dd, 5.2,J= 5.2, 49 49 798.6 A A 12.8 Hz), 12.8 Hz),6.866.86(1H, (1H,d, d, J =J= 9.29.2Hz),Hz),7.007.00 (1H, (1H, dd, J dd, = 8.4,J=2.4 8.4,Hz),2.47.10 Hz),(1H, 7.10 dd,(1H, dd, JJ= 10.0,2.0 = 10.0, 2.0Hz), Hz),7.21 7.21(1H,(1H, d, Jd,= J= 2.4 2.4 Hz),Hz), 7.25 7.25 (1H, d, (1H, J = d, J=Hz), 10.0 10.07.36Hz), (1H,7.36 (1H, s), 7.62 (1H, d, J= 9.2 Hz), 7.90 (1H, d, J= 8.8 Hz), 7.95 (1H, dd, J= s), 7.62 (1H, d, J = 9.2 Hz), 7.90 (1H, d, J = 8.8 Hz), 7.95 (1H, dd, J = 9.2, 2.49.2, 2.4

Hz), 8.62 Hz), 8.62(1H,(1H,d, d,J =J= 2.42.4 Hz),Hz), 11.1011.10 (1H, (1H, s). s). ¹H NMR (400 MHz, DMSO-d) 1.13 6 H NMR (400 MHz, DMSO-d ) 6 1.13 (6H, s), 1.22 (6H, s), 1.79-1.81 (6H, s), 1.22 (6H, s), 1.79-1.81 (3H, m), (3H, m), 2.09-2.15 (1H, 2.09-2.15 (1H, m), m), 2.19-2.21 2.19-2.21 (2H, (2H, m), m), 2.49-2.50 2.49-2.50(7H, (7H,m), m),2.60-2.67 2.60-2.67(1H, (1H,m), m), 2.76-2.92 (3H, m), 3.22-3.26 (4H, m), 3.86 (2H, d, J= 12.8 Hz), 4.05 (1H, d, 2.76-2.92 (3H, m), 3.22-3.26 (4H, m), 3.86 (2H, d, J = 12.8 Hz), 4.05 (1H, d, J J 808.6 808.6 50 50 A A 9.2 Hz), == 9.2 Hz),4.32 4.32(1H,(1H,s),s), 5.235.23 (1H,(1H,dd, Jdd, J= 12.4, = 12.4, 5.2 Hz),5.2 6.12 Hz),(1H, 6.12s),(1H,6.70s), 6.70 (1H, (1H, A A dd, JJ= dd, 8.0, 1.6 = 8.0, 1.6 Hz), Hz), 6.95 (2H, d, J= 9.2 Hz), 7.00 (1H, dd, J= 8.8, 2.4 Hz), 6.95 (2H, d, J = 9.2 Hz), 7.00 (1H, dd, J = 8.8, 2.4 Hz),

7.21 (1H, 7.21 (1H,d,d,J J== 2.42.4Hz), Hz),7.487.48 (1H,(1H,d, J d, J= Hz), = 8.8 8.8 Hz), 7.72t,(3H, 7.72 (3H, t, J= J = 8.4 Hz),8.47.91 Hz), 7.91 (1H, (1H, d, d, JJ== 8.88.8Hz), Hz),11.0411.04 (1H,(1H, s). s).

1H NMR 1H NMR (300(300MHz,MHz, DMSO-d6) DMSO-d6) 11.07 (s, 6 11.07 1H), (s, 8.571H),(s,8.57 1H),(s,7.87 1H),(d,7.87 J =(d, 8.7J = 8.7 Hz, 1H), Hz, 1H),7.70 7.70(d,(d, J =J= 8.68.6 Hz,Hz,2H),2H),7.44 7.44 (d, J(d, J =Hz, = 9.1 9.1 1H), Hz, 7.29 1H),(s,7.291H),(s, 7.171H), (d,7.17 (d,

J= 2.2 Hz, 1H), 7.02-6.87 (m, 3H), 5.07 (dd, J= 12.8, 5.3 Hz, 1H), 4.29 (s, 1H), 835.59 J = 2.2 Hz, 1H), 7.02-6.87 (m, 3H), 5.07 (dd, J = 12.8, 5.3 Hz, 1H), 4.29 (s, 1H), 51 51 835.59 A A A A 4.02 (d, 4.02 (d, JJ= 9.1Hz, = 9.1 Hz,1H),1H), 3.88-3.70 3.88-3.70 (m, 3.29 (m, 5H), 5H), (br3.29 S, (br 5H),s,2.95-2.65 5H), 2.95-2.65 (m, 3H), (m, 3H),

2.59-2.41 (m, 6H), 2.00 (m, 1H), 1.73 (d, J= 12.8 Hz, 2H), 1.45 (br, 2.59-2.41 (m, 6H), 2.00 (m, 1H), 1.73 (d, J = 12.8 Hz, 2H), 1.45 (br, 3H), 1.17 3H), 1.17

(s, 6H), (s, 6H), 1.091.09(s, (s, 6H) 6H) ¹HH NMR (400 MHz, d6-DMSO): 6 11.12 (s, 1H),(s,8.90 NMR (400 MHz, d6-DMSO): 11.12 (s, 1H), 8.90 1H), (s,7.91-7.89 1H), 7.91-7.89 (d, (d, J=8.4Hz, 1H), 7.74-7.72 (d, J=7.6Hz, 2H), 7.49-7.47 (d, J=8.8Hz, J=8.4Hz, 1H), 7.74-7.72 (d, J=7.6Hz, 2H), 7.49-7.47 (d, J=8.8Hz, 1H),1H), 7.20 7.20 (s, (s, 836.59 836.59 52 52 A B B 1H), 6.99-6.94 6.99-6.94 (m, (m, 3H), 3H), 5.16-5.13 5.16-5.13 (m, (m,1H),1H),4.32 4.32(s, (s, 1H), 4.06-3.83 (m, 1H), 4.06-3.83 (m, 7H), 7H), A 2.88-2.57 (m, 2.88-2.57 (m, 5H), 5H), 2.39-2.33 2.39-2.33 (m, (m,2H), 2H),2.07-2.01 2.07-2.01(m, 1H),1.78-1.75 (m,1H), 1.78-1.75(m,(m, 2H), 2H), 1.54-1.35 (m, 1.54-1.35 (m, 3H),3H), 1.21 1.21 (m, (m, 8H), 8H),1.121.12(s, (s, 6H) 6H)

*AR DC *AR DC (nM) 5 o (nM) A<1;1<=B<10; A<1; 1<=B<10;10<=C<100; 10<=C<100;D>=100 D>=100 **ARDmax(%) **AR Dmax(%)A>=75; 50<=B<75; A>=75;50<=B<75; C<50 C<50

427

[0955]

[0955] Table 7. Characterization Table 7. of Exemplary Characterization of BRaf Exemplary BRaf PROTACs PROTACs

Ex. # BRaf BRaf Dma, MH+ Ex. # DCa5 Dmax** MH+ NMRTranscript NMR Transcript DC50* 1H NMR 1H NMR (400 (400 MHz, MHz, DMSO-d6): DMSO-d6): 6 11.03 11.03 (s, 1H), (s, 1H),(s, 10.87 10.87 1H),(s,8.72 1H),(s, 8.72 (s, 8.57 1H), 1H),(m, 8.572H), (m, 2H), 783.51 7.83 (d, J = 8.4 Hz, 2H), 7.62-7.60 (m, 2H), 7.56 (d, J = 8.4J= 7.83 (d, J= 8.4 Hz, 2H), 7.62-7.60 (m, 2H), 7.56 (d, Hz,8.4 Hz, 2H), 2H), 7.50-7.41 7.50-7.41 (m, (m, 6H), 7.22 6H), 7.22 39 39 C B B 783.51 C (d, J = 8.0 Hz, 2H), 7.17-7.15(m, 3H), 5.07-5.03 (m, 1H), 3.73 (m, 8H), 3.01 (s, 2H), 2.83-2.81 (d, J = 8.0 Hz, 2H), 7.17-7.15(m, 3H), 5.07-5.03 (m, 1H), 3.73 (m, 8H), 3.01 (s, 2H), 2.83-2.81

(m, 3H), (m, 2.67 (s, 3H), 2.67 (s, 2H), 2H), 2.03-2.00 2.03-2.00 (m, (m, 1H) 1H) 1H NMR 1H NMR (400(400 MHz, MHz, DMSO-d6): DMSO-d6): 6 11.15 11.15 (bs, 1H),(bs, 8.721H), (s,8.72 1H),(s,8.72 1H),(s,8.72 1H),(s, 8.57 1H),(d,8.57J =(d, J= A 721.48 5.6Hz, 2H), 7.83 (d, J = 8.8 Hz, 2H), 7.77 (s, 1H), 7.49-7.57 (m, 4H),(m, 5.6Hz, 2H), 7.83 (d, J = 8.8 Hz, 2H), 7.77 (s, 1H), 7.49-7.57 7.41 4H), (s, 7.41 1H), (s, 1H), 7.17-7.30 7.17-7.30 40 40 C 721.48 C A (m, 3H), (m, 3H), 5.09-5.13 5.09-5.13 (m, 1H), 3.20-3.35 (m, 1H), 3.20-3.35 (m,(m,8H), 8H), 2.80-3.12 2.80-3.12 (m, (m,6H), 6H),2.52-2.75 2.52-2.75(m, (m,3H), 3H),1.90- 1.90 2.12 (m, 2.12 (m, 2H) 2H) 1H NMR 1H NMR (400(400 MHz, MHz, DMSO-d6): DMSO-d6): 11.04 (s,611.04 1H), (s, 1H),(s, 10.88 10.88 1H),(s,8.691H),(s,8.69 1H),(s, 8.57 1H),(d,8.57J =(d,4.8 J= 4.8 A 783.51 Hz, 2H), 7.87 (d, J = 8.4 Hz, 1H), 7.76 (d, J = 8.8 Hz, 2H), 7.40-7.56 Hz, 2H), 7.87 (d, J= 8.4 Hz, 1H), 7.76 (d, J= 8.8 Hz, 2H), 7.40-7.56 (m, 10H), (m, 7.22 10H), (d, J 7.22 = 4.8 (d, J= 4.8 41 41 C 783.51 C A Hz, 1H), 7.03 (d, J= 9.2 Hz, 2H), 5.00-5.05 (m, 1H), 3.02 (m, 9H), 2.83 (t, J= 6.8 Hz, 2H), Hz, 1H), 7.03 (d, J = 9.2 Hz, 2H), 5.00-5.05 (m, 1H), 3.02 (m, 9H), 2.83 (t, J = 6.8 Hz, 2H),

1.99-2.01 (m, 1.99-2.01 (m, 3H) 3H) 1H NMR (400 MHz, DMSO-d6): 6 11.09 (s, 1H), 10.89 1H NMR (400 MHz, DMSO-d6): 11.09 (s, 1H), 10.89 (s, 1H),(s,8.72 1H),(s, 8.72 1H), (s, 8.58-8.57 1H), 8.58-8.57(m, (m, 2H), 7.83 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 7.6 Hz, 1H), 7.56 (d, J = 7.6 Hz, 1H), 7.50-7.41 (m, 42 42 C B 721.48 721.48 2H), 7.83 (d, J= 8.0 Hz, 2H), 7.73 (d, J= 7.6 Hz, 1H), 7.56 (d, J= 7.6 Hz, 1H), 7.50-7.41 (m, C B 4H), 7.23-7.17 4H), 7.23-7.17 (m, (m, 3H), 3H), 5.13-5.09 5.13-5.09 (m, 1H),3.61-3.42 (m,1H), 3.61-3.42(m, (m,8H), 8H),3.04-2.97 3.04-2.97(m, (m,2H), 2H),2.93-2.82 2.93-2.82 (m, 3H), 2.62-2.56 (m, 3H), 2.62-2.56 (m,(m, 5H), 5H), 2.08-2.00 2.08-2.00 (m, (m, 1H)1H) *BRaf DC *BRaf o (nM)A<1; DC 5(nM) A<1;1<=B<10; 1<=B<10;10<=C<100; 10<=C<100; D>=100 D>=100 **BRaf Dmax(%) A>=75; **BRaf Dmax(%) A>=75;50<=B<75; 50<=B<75;C<50 C<50

[0956] 8. 8. Table

[0956] Table Characterization Exemplary BRD4 of Exemplary Characterization of PROTACs BRD4 PROTACs

Ex. Ex. BRD4 BRD4 BRD4 BRD4 Observed NMR # # DC5o* DC50* Dmax** Dmax** [M+H]+

[M+H]+ NMR H NMR ¹H NMR (400 (400 MHz, MHz, CHLOROFORM-d) CHLOROFORM-d) d 9.037.45 d 9.03 (s, 1H), 1H), J7.45 (s, (dd, (dd, J= = 8.71, 8.71, 13.21 Hz, 13.21 4H), Hz, 4H), 43 43 D B B 895.22 895.22 7.31 -- 7.37 7.31 7.37(m, (m,3H), 3H), 7.24 7.24 (d, (d, J = J= 7.247.24 Hz, Hz, 1H),6.84 1H), 6.84 (d, J =(d, J=Hz, 9.00 9.00 Hz, 2H), 2H), 6.78 (d, 6.78 (d, J = 8.02 J= 8.02 D Hz, 1H),6.75 Hz, 1H), 6.75 (br. 1H),4.66 (br.S.,s.,1H), 4.66 - 4.73 - 4.73 (m, (m, 2H),2H), 4.20 4.20 (d, J (d, J= 2.74 Hz, 1H), 4.07 - 4.12 (m, = 2.74 Hz, 1H), 4.07 - 4.12 (m,

428

2H), 3.80- -3.90 2H),3.80 3.90(m,(m, 3H), 3H), 3.643.64 - 3.77 - 3.77 (m, 10H), 1OH), (m, 3.52 - 3.52 - 3.58 3.58 (m, 1H),(m, 1H), 3.35 - 3.42 (m,- 3H), 3.35 3.42 (m, 3H), 2.68 (br. 2.68 (br. S., s., 3H), 2.52- -2.59 3H), 2.52 2.59(m,(m, 2H), 2H), 2.412.41 (s, 3H), (s, 3H), - 2.08- 2.08 2.02 2.02 (m,1.69 (m, 2H), 2H),(s,1.69 3H),(s, 3H), 1.26 1.26 (s, 3H). '(HE 's)

1H NMR 1H NMR (400 (400 MHz, MHz, METHANOL-d4) METHANOL-d4) d 7.60 - 7.65 (m, - 1H), d 7.60 (m, 1H), 7.65 7.30 (m,- 8H), 7.30 - 7.47 7.47 6.82 (m, 8H), - 6.82 6.87 (m, 6.87 (m,2H), 2H),5.24 5.24 (dd, (dd, J = J= 5.67, 5.67, 10.76 10.76 Hz, 1H), 1H), Hz, 4.69 (ddd, 4.69J (ddd, J= = 2.84, 2.84, 5.62, 5.62, 8.56 Hz, 1H), 8.56 Hz, 1H), 4.26 - 4.31 (m, 2H), 4.02 - 4.07 (m, 2H), 3.91 - 3.96 (m, 2H), 3.77 - 3.81 (m, 2H), 3.70 4.26 - 4.31 (m, 2H), 4.02 - 4.07 (m, 2H), 3.91 - 3.96 (m, 2H), 3.77 - 3.81 (m, 2H), 3.70 - 44 44 D CI C 937.19 61.7.9 ) 3.74 (m, 3.74 (m,2H), 2H), 3.63 3.63 - 3.69 - 3.69 (m, (m, 6H), 6H), 3.53 - 3.61- (m, 3.53 3.611H), 1H),- 3.49 (m,3.43 - 3.49 3.43(m, 2H), (m, 2.81 2H), 2.81 (dt, J = (dt, J = 4.60,14.33 4.60, Hz, 14.33 Hz, 2H), 2H), 2.702.70 (s, 6H), (s, 6H), 2.43 2.43 (s, 3H), (s, 3H), 2.20 -(m, 2.13 -2.13 2.20 1H), (m,1.68 1H), (s, 1.68 2H), (s, 1.262H), - 1.26 2023248067 1.29 (m, 2H). 1.29

H NMR ¹H NMR (400 (400 MHz, MHz, METHANOL-d4) METHANOL-d4) d 8.55 (s,d1H), (s, 1H), 8.55 7.96 - 8.00 7.96(m,- 8.00 1H), (m, 7.361H), - 7.50 7.36(m, - 7.50 (m, 6H), 7.03- -7.09 6H), 7.03 7.09(m,(m, 2H), 2H), 6.876.87 (dd, (dd, J= 3.03, J = 3.03, 9.102H), 9.10 Hz, Hz,5.22 2H), 5.22 (td, J = (td, J=10.91 5.40, 5.40,Hz,10.91 1H), Hz, 1H), 4.70 - 4.74 (m, 1H), 4.22 (d, J = 3.33 Hz, 2H), 4.10 (d, J = 4.30 Hz, 2H), 3.85 - 3.91 (m, 45 45 C A 937.19 617.19 4.70 - 4.74 (m, 1H), 4.22 (d, J= 3.33 Hz, 2H), 4.10 (d, J= 4.30 Hz, 2H), 3.85 - 3.91 (m, 0 V 2H),3.79 2H), 3.79- -3.84 3.84(m,(m, 2H), 2H), 3.643.64 - 3.71 - 3.71 (m, 3.55 (m, 7H), 7H),- 3.55 - 3.64 3.64 (m, 2H),(m, 2H), 3.42 3.42 - 3.50 (m,- 2H), 3.50 2.71 (m, 2H), 2.71 (s, 3H), (s, 2.66(d, 3H), 2.66 (d,JJ= 3.33Hz,Hz, = 3.33 2H), 2H), 2.442.44 (d, J(d,= J = 3.33 3.33 Hz, 1.89 Hz, 3H), 3H),(s,1.89 3H),(s,1.68 3H), (d,1.68 (d, J= 3.33 J = 3.33

Hz, 2H), 1.29 (br. s., 3H). Hz, 2H), 1.29 (br. S., 3H).

*BRD4DC *BRD4 DC5(nM) o (nM) A<1; A<1; 1<=B<10; 1<=B<10; 10<=C<100;D>=100 10<=C<100; D>=100 **BRD4 Dmax(%) *BRD4 D A>=75;50<=B<75; (%) A>=75; 50<=B<75; C<50 C<50

429

[0957]

[0957] 5. Industrial 5. Industrialapplicability applicability

[0958] A novel

[0958] A novel bifunctionalmolecule, bifunctional molecule,which which containsa BRD4 contains a BRD4 or androgen or an an androgen receptor receptor

recruiting moiety recruiting and an moiety and an E3 E3Ligase LigaseCereblon Cereblonrecruiting recruitingmoiety, moiety,through through PROTAC PROTAC technology technology is is described. TheThe described. bifunctional bifunctional molecules molecules ofpresent of the the present disclosure disclosure actively actively degrades degrades BRD4, BRD4, leading to significant leading to significant and and persistent persistentdownstream MYC downstream MYC suppression suppression andand robust robust cellular cellular

proliferation suppression proliferation suppressionand andapoptosis apoptosisinduction. PROTAC induction. mediated protein PROTAC mediated protein degradation degradation provides aa promising provides promisingstrategy strategy in in targeting targeting the the "undruggable" pathologicalproteins "undruggable" pathological proteins by bytraditional traditional approaches. approaches.

The contents

[0959] The contents

[0959] all references, of allofreferences, patents, patents, pending patent patent pending applications applications and published and published

patents, cited patents, cited throughout throughout this this application application are hereby are hereby expressly expressly incorporated incorporated by reference. by reference.

[0960] ThoseThose

[0960] skilled skilled in the art will the will in art or beorable recognize, recognize, be to usingusing to ascertain ableascertain no more more no than than routine experimentation, routine experimentation, many equivalents to many equivalents the specific to the specific embodiments of the embodiments of the invention invention described herein. described herein. Such Suchequivalents equivalentsare are intended intendedtoto be be encompassed encompassedby by thethe following following claims. claims. It Itisis

understoodthat understood that the the detailed detailed examples examplesand and embodiments embodiments described described hereinherein are given are given by wayby ofway of example forillustrative example for illustrative purposes purposesonly, only,andand areare in way in no no way considered considered to be limiting to be limiting to the to the

invention. Variousmodifications invention. Various modificationsororchanges changesininlight lightthereof thereof will will be be suggested suggested to to persons persons skilled skilled in the the art art and and are are included included within the the spirit spirit and and purview of this purview of this application application and and are are considered considered within the within the scope scope of of the the appended claims.ForFor appended claims. example, example, the the relativequantities relative quantitiesofofthe theingredients ingredients maybebevaried may variedto to optimize optimize the the desired desired effects, effects, additional additional ingredients ingredients may may be added, be added, and/or and/or similar similar ingredients ingredients may be substituted may be substituted for for one one or or more moreofofthe the ingredients ingredients described. described. Additional Additional advantageousfeatures advantageous featuresand andfunctionalities functionalities associated associated with the systems, methods, with the methods, and andprocesses processesofof the present disclosure the disclosure will willbe beapparent apparent from from the the appended claims. Moreover, appended claims. Moreover, those those skilledininthe skilled the art will art will recognize, or be able recognize, or able to to ascertain ascertain using using no nomore morethan than routine routine experimentation, experimentation, manymany

equivalents to to the the specific specific embodiments embodiments ofofthe the invention invention described describedherein. herein. Such Such equivalentsareare equivalents

intended to be intended to be encompassed encompassed byby thefollowing the followingclaims. claims.

430

Claims (24)

The claims defining the invention are as follows:

1. A compound selected from the group consisting of Ex. # Chemical Structure

1 2023248067

2

3

4

5

8 7 6

11 10

16 15 14 13 12

22 21 20 19 18

28 27 26 25 24

2023248067

31

or a pharmaceutically acceptable salt thereof.

2. A composition comprising an effective amount of the compound of claim 1, and a pharmaceutically acceptable carrier.

3. The composition of claim 2, wherein the composition further comprises at least one of an additional bioactive agent or another compound of claim 1.

4. The composition of claim 3, wherein the additional bioactive agent is anti-cancer agent, an anti-neurodegenerative agent, an antimicrobial agent, an antiviral agent, an anti-HIV agent, or an antifungal agent.

5. Use of an effective amount of a compound of claim 1 for the manufacture of a medicament for treating a disease or disorder in a subject, wherein the disease or disorder is associated with the accumulation and/or aggregation of the target protein.

6. The use of claim 5, wherein the disease or disorder is selected from the group consisting of asthma, autoimmune diseases such as multiple sclerosis, various cancers, ciliopathies, cleft palate, diabetes, heart disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease,

Coeliac disease, Charcot–Marie–Tooth disease, Cystic fibrosis, Duchenne muscular dystrophy, Haemochromatosis, Haemophilia, Klinefelter's syndrome, Neurofibromatosis, Phenylketonuria, Polycystic kidney disease, (PKD1) or 4 (PKD2) Prader–Willi syndrome, Sickle-cell disease, Tay–Sachs disease, Turner syndrome.

7. The use of claim 5, wherein the disease or disorder is selected from the group consisting of Alzheimer's disease, Amyotrophic lateral sclerosis (Lou Gehrig’s disease), Anorexia nervosa, 2023248067

Anxiety disorder, Atherosclerosis, Attention deficit hyperactivity disorder, Autism, Bipolar disorder, Chronic fatigue syndrome, Chronic obstructive pulmonary disease, Crohn's disease, Coronary heart disease, Dementia, Depression, Diabetes mellitus type 1, Diabetes mellitus type 2, Epilepsy, Guillain–Barré syndrome, Irritable bowel syndrome, Lupus, Metabolic syndrome, Multiple sclerosis, Myocardial infarction, Obesity, Obsessive–compulsive disorder, Panic disorder, Parkinson's disease, Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia, Stroke, Thromboangiitis obliterans, Tourette syndrome, Vasculitis.

8. The use of claim 5, wherein the disease or disorder is selected from the group consisting of aceruloplasminemia, Achondrogenesis type II, achondroplasia, Acrocephaly, Gaucher disease type 2, acute intermittent porphyria, Canavan disease, Adenomatous Polyposis Coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, Adrenogenital syndrome, Adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, Alkaptonuria, Alexander disease, Alkaptonuric ochronosis, alpha 1-antitrypsin deficiency, alpha-1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis Alström syndrome, Alexander disease, Amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, Anemia Angiokeratoma Corporis Diffusum, Angiomatosis retinae (von Hippel–Lindau disease) Apert syndrome, Arachnodactyly (Marfan syndrome), Stickler syndrome, Arthrochalasis multiplex congenital (Ehlers–Danlos syndrome#arthrochalasia type) ataxia telangiectasia, Rett syndrome, primary pulmonary hypertension, Sandhoff disease, neurofibromatosis type II, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever, familial, Benjamin syndrome, beta-thalassemia, Bilateral Acoustic Neurofibromatosis (neurofibromatosis type II), factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome (Turner syndrome), Bourneville disease (tuberous sclerosis), prion disease, Birt– Hogg–Dubé syndrome, Brittle bone disease (osteogenesis imperfecta), Broad Thumb-Hallux

syndrome (Rubinstein-Taybi syndrome), Bronze Diabetes/Bronzed Cirrhosis (hemochromatosis), Bulbospinal muscular atrophy (Kennedy's disease), Burger-Grutz syndrome (lipoprotein lipase deficiency), CGD Chronic granulomatous disorder, Campomelic dysplasia, biotinidase deficiency, Cardiomyopathy (Noonan syndrome), Cri du chat, CAVD (congenital absence of the vas deferens), Caylor cardiofacial syndrome (CBAVD), CEP (congenital erythropoietic porphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophy syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, Lesch- 2023248067

Nyhan syndrome, galactosemia, Ehlers–Danlos syndrome, Thanatophoric dysplasia, Coffin- Lowry syndrome, Cockayne syndrome, (familial adenomatous polyposis), Congenital erythropoietic porphyria, Congenital heart disease, Methemoglobinemia/Congenital methaemoglobinaemia, achondroplasia, X-linked sideroblastic anemia, Connective tissue disease, Conotruncal anomaly face syndrome, Cooley's Anemia (beta-thalassemia), Copper storage disease (Wilson's disease), Copper transport disease (Menkes disease), hereditary coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome), Creutzfeldt-Jakob disease (prion disease), Cockayne syndrome, Cowden syndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher syndrome, Degenerative nerve diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disabilities, distal spinal muscular atrophy, type V, androgen insensitivity syndrome, Diffuse Globoid Body Sclerosis (Krabbe disease), Di George's syndrome, Dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, Dwarfism, erythropoietic protoporphyria Erythroid 5-aminolevulinate synthetase deficiency, Erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Friedreich's ataxia,, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure sensitive neuropathy, primary pulmonary hypertension (PPH), Fibrocystic disease of the pancreas, fragile X syndrome, galactosemia, genetic brain disorders, Giant cell hepatitis (Neonatal hemochromatosis), Gronblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther disease (congenital erythropoietic porphyria), haemochromatosis, Hallgren syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease (von Hippel-Lindau disease), Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), Hyperandrogenism, Hypochondroplasia, Hypochromic anemia, Immune system disorders, including X-linked severe combined immunodeficiency, Insley-Astley syndrome,

Kennedy’s syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome, Kidney diseases, including hyperoxaluria, Klinefelter's syndrome, Kniest dysplasia, Lacunar dementia,Langer-Saldino achondrogenesis, ataxia telangiectasia, Lynch syndrome, Lysyl-hydroxylase deficiency, Machado-Joseph disease, Metabolic disorders, including Kniest dysplasia, Marfan syndrome, Movement disorders, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome, Multiple neurofibromatosis, Nance-Insley syndrome, Nance- Sweeney chondrodysplasia, Niemann–Pick disease, Noack syndrome (Pfeiffer syndrome), 2023248067

Osler-Weber-Rendu disease, Peutz-Jeghers syndrome, Polycystic kidney disease, polyostotic fibrous dysplasia (McCune–Albright syndrome), Peutz-Jeghers syndrome, Prader-Labhart- Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, prion disease, progeria (Hutchinson Gilford Progeria Syndrome), progressive chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular atrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb (retinoblastoma), Recklinghausen disease (neurofibromatosis type I), Recurrent polyserositis, Retinal disorders, Retinoblastoma, Rett syndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levy syndrome, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome, sclerosis tuberose (tuberous sclerosis), SDAT, SED congenital (spondyloepiphyseal dysplasia congenita), SED Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type (spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome, Skin pigmentation disorders, Smith-Lemli-Opitz syndrome, South- African genetic porphyria (variegate porphyria), infantile-onset ascending hereditary spastic paralysis, Speech and communication disorders, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, Thyroid disease, Tomaculous neuropathy (hereditary neuropathy with liability to pressure palsies), Treacher Collins syndrome, Triplo X syndrome ( triple X syndrome), Trisomy 21 (Down syndrome), Trisomy X, VHL syndrome (von Hippel-Lindau disease), Vision impairment and blindness (Alström syndrome), Vrolik disease, Waardenburg syndrome, Warburg Sjo Fledelius Syndrome, Weissenbacher- Zweymüller syndrome, Wolf–Hirschhorn syndrome, Wolff Periodic disease, Weissenbacher- Zweymüller syndrome and Xeroderma pigmentosum.

9. The use of any one of claims 5 to 8, further comprising an additional bioactive agent.

10. The use of claim 9, wherein the additional bioactive agent is at least one of an anti-cancer agent, an anti-neurodegenerative agent, an antimicrobial agent, an antiviral agent, an anti-HIV agent, an antifungal agent, or a combination thereof. 2023248067

11. The use of claim 10, wherein said anticancer agent is selected from the group consisting of everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK- 1 modulator, a Bcl-2 inhibitor, an HDAC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AKT inhibitor, an mTORC1/2 inhibitor, a JAK/STAT inhibitor, a checkpoint-1 or 2 inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601 , ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001 , IPdR1 KRX-0402, lucanthone, LY 317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311 , romidepsin, ADS- 100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, liposomal doxorubicin, 5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709, seliciclib; PD0325901 , AZD-6244, capecitabine, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11 , CHIR-258,); 3-[5-(methylsulfonylpiperadinemethyl)- indolylj- quinolone, vatalanib, AG-013736, AVE-0005, the acetate salt of [D- Ser(Bu t ) 6 ,Azgly 10 ] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t )-Leu-Arg-Pro- Azgly-NH 2 acetate [C59H84N18Oi4 - (C2H4O2)X where x = 1 to 2.4], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, erbitux, EKB-569, PKI-166, GW-572016,

Ionafarnib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951 , aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gleevac, gemcitabine, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, 2023248067

mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291 , squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox,gefitinib, bortezimib, paclitaxel, cremophor-free paclitaxel, docetaxel, epithilone B, BMS- 247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA- 923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR- 3339, ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)- rapamycin, temsirolimus, AP-23573, RAD001 , ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L- asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11 , dexrazoxane, alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2, megestrol, immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, an NK-1 receptor antagonists, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol,

dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa, darbepoetin alfa and mixtures thereof.

12. Use of an effective amount of a compound of claim 1 for the manufacture of a medicament for inducing degradation of a target protein, wherein the compound effectuates degradation of the target protein. 2023248067

13. Use of an effective amount of a compound of claim 1 for the manufacture of a medicament for treating cancer, wherein the compound effectuates for the treatment or alleviation of at least one symptom of cancer in the patient.

14. The use of claim 13, wherein the cancer is squamous-cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; multiple myeloma, sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor or teratocarcinomas, T-lineage Acute lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosome positive ALL and Philadelphia chromosome positive CML.

15. A method for treating a disease or disorder in a subject, wherein the disease or disorder is associated with the accumulation and/or aggregation of the target protein, said method comprising administering to the subject an effective amount of a compound of claim 1, or a pharmaceutically acceptable thereof.

16. The method of claim 15, wherein the disease or disorder is selected from the group consisting of asthma, autoimmune diseases such as multiple sclerosis, various cancers, ciliopathies, cleft palate, diabetes, heart disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, Coeliac disease, Charcot–Marie–Tooth disease, Cystic fibrosis, Duchenne muscular dystrophy, Haemochromatosis, Haemophilia, Klinefelter's syndrome, 2023248067

Neurofibromatosis, Phenylketonuria, Polycystic kidney disease, (PKD1) or 4 (PKD2) Prader– Willi syndrome, Sickle-cell disease, Tay–Sachs disease, Turner syndrome.

17. The method of claim 15, wherein the disease or disorder is selected from the group consisting of Alzheimer's disease, Amyotrophic lateral sclerosis (Lou Gehrig’s disease), Anorexia nervosa, Anxiety disorder, Atherosclerosis, Attention deficit hyperactivity disorder, Autism, Bipolar disorder, Chronic fatigue syndrome, Chronic obstructive pulmonary disease, Crohn's disease, Coronary heart disease, Dementia, Depression, Diabetes mellitus type 1, Diabetes mellitus type 2, Epilepsy, Guillain–Barré syndrome, Irritable bowel syndrome, Lupus, Metabolic syndrome, Multiple sclerosis, Myocardial infarction, Obesity, Obsessive–compulsive disorder, Panic disorder, Parkinson's disease, Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia, Stroke, Thromboangiitis obliterans, Tourette syndrome, Vasculitis.

18. The method of claim 15, wherein the disease or disorder is selected from the group consisting of aceruloplasminemia, Achondrogenesis type II, achondroplasia, Acrocephaly, Gaucher disease type 2, acute intermittent porphyria, Canavan disease, Adenomatous Polyposis Coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, Adrenogenital syndrome, Adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, Alkaptonuria, Alexander disease, Alkaptonuric ochronosis, alpha 1-antitrypsin deficiency, alpha-1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis Alström syndrome, Alexander disease, Amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, Anemia Angiokeratoma Corporis Diffusum, Angiomatosis retinae (von Hippel–Lindau disease) Apert syndrome, Arachnodactyly (Marfan syndrome), Stickler syndrome, Arthrochalasis multiplex congenital (Ehlers–Danlos syndrome#arthrochalasia type) ataxia telangiectasia, Rett syndrome, primary pulmonary hypertension, Sandhoff disease, neurofibromatosis type II, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever,

familial, Benjamin syndrome, beta-thalassemia, Bilateral Acoustic Neurofibromatosis (neurofibromatosis type II), factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome (Turner syndrome), Bourneville disease (tuberous sclerosis), prion disease, Birt– Hogg–Dubé syndrome, Brittle bone disease (osteogenesis imperfecta), Broad Thumb-Hallux syndrome (Rubinstein-Taybi syndrome), Bronze Diabetes/Bronzed Cirrhosis (hemochromatosis), Bulbospinal muscular atrophy (Kennedy's disease), Burger-Grutz 2023248067

syndrome (lipoprotein lipase deficiency), CGD Chronic granulomatous disorder, Campomelic dysplasia, biotinidase deficiency, Cardiomyopathy (Noonan syndrome), Cri du chat, CAVD (congenital absence of the vas deferens), Caylor cardiofacial syndrome (CBAVD), CEP (congenital erythropoietic porphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophy syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, Lesch- Nyhan syndrome, galactosemia, Ehlers–Danlos syndrome, Thanatophoric dysplasia, Coffin- Lowry syndrome, Cockayne syndrome, (familial adenomatous polyposis), Congenital erythropoietic porphyria, Congenital heart disease, Methemoglobinemia/Congenital methaemoglobinaemia, achondroplasia, X-linked sideroblastic anemia, Connective tissue disease, Conotruncal anomaly face syndrome, Cooley's Anemia (beta-thalassemia), Copper storage disease (Wilson's disease), Copper transport disease (Menkes disease), hereditary coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome), Creutzfeldt-Jakob disease (prion disease), Cockayne syndrome, Cowden syndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher syndrome, Degenerative nerve diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disabilities, distal spinal muscular atrophy, type V, androgen insensitivity syndrome, Diffuse Globoid Body Sclerosis (Krabbe disease), Di George's syndrome, Dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, Dwarfism, erythropoietic protoporphyria Erythroid 5-aminolevulinate synthetase deficiency, Erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Friedreich's ataxia,, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure sensitive neuropathy, primary pulmonary hypertension (PPH), Fibrocystic disease of the pancreas, fragile X syndrome, galactosemia, genetic brain disorders, Giant cell hepatitis (Neonatal hemochromatosis), Gronblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther

disease (congenital erythropoietic porphyria), haemochromatosis, Hallgren syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease (von Hippel-Lindau disease), Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), Hyperandrogenism, Hypochondroplasia, Hypochromic anemia, Immune system disorders, including X-linked severe combined immunodeficiency, Insley-Astley syndrome, Kennedy’s syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome, Kidney diseases, including hyperoxaluria, Klinefelter's syndrome, 2023248067

Kniest dysplasia, Lacunar dementia, Langer-Saldino achondrogenesis, ataxia telangiectasia, Lynch syndrome, Lysyl-hydroxylase deficiency, Machado-Joseph disease, Metabolic disorders, including Kniest dysplasia, Marfan syndrome, Movement disorders, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome, Multiple neurofibromatosis, Nance-Insley syndrome, Nance- Sweeney chondrodysplasia, Niemann–Pick disease, Noack syndrome (Pfeiffer syndrome), Osler-Weber-Rendu disease, Peutz-Jeghers syndrome, Polycystic kidney disease, polyostotic fibrous dysplasia (McCune–Albright syndrome), Peutz-Jeghers syndrome, Prader-Labhart- Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, prion disease, progeria (Hutchinson Gilford Progeria Syndrome), progressive chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular atrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb (retinoblastoma), Recklinghausen disease (neurofibromatosis type I), Recurrent polyserositis, Retinal disorders, Retinoblastoma, Rett syndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levy syndrome, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome, sclerosis tuberose (tuberous sclerosis), SDAT, SED congenital (spondyloepiphyseal dysplasia congenita), SED Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type (spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome, Skin pigmentation disorders, Smith-Lemli-Opitz syndrome, South- African genetic porphyria (variegate porphyria), infantile-onset ascending hereditary spastic paralysis, Speech and communication disorders, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, Thyroid disease, Tomaculous neuropathy (hereditary neuropathy with liability to pressure palsies), Treacher Collins syndrome, Triplo X

syndrome ( triple X syndrome), Trisomy 21 (Down syndrome), Trisomy X, VHL syndrome (von Hippel-Lindau disease), Vision impairment and blindness (Alström syndrome), Vrolik disease, Waardenburg syndrome, Warburg Sjo Fledelius Syndrome, Weissenbacher- Zweymüller syndrome, Wolf–Hirschhorn syndrome, Wolff Periodic disease, Weissenbacher- Zweymüller syndrome and Xeroderma pigmentosum.

19. The method of any one of claims 15 to 18, the method further comprising administering 2023248067

to the subject an additional bioactive agent.

20. The method of claim 19, wherein the additional bioactive agent is at least one of an anti- cancer agent, an anti-neurodegenerative agent, an antimicrobial agent, an antiviral agent, an anti- HIV agent, an antifungal agent, or a combination thereof.

21. The method of claim 20, wherein said anticancer agent is selected from the group consisting of everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA- 739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HDAC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AKT inhibitor, an mTORC1/2 inhibitor, a JAK/STAT inhibitor, a checkpoint-1 or 2 inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601 , ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001 , IPdR1 KRX-0402, lucanthone, LY 317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311 , romidepsin, ADS- 100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, liposomal doxorubicin, 5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709, seliciclib; PD0325901 , AZD-6244, capecitabine, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11 , CHIR-258,); 3-[5-

(methylsulfonylpiperadinemethyl)- indolylj-quinolone, vatalanib, AG-013736, AVE-0005, the acetate salt of [D- Ser(Bu t ) 6 ,Azgly 10 ] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t )-Leu-Arg- Pro- Azgly-NH 2 acetate [C59H84N18Oi4 -(C2H4O2)X where x = 1 to 2.4], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, erbitux, EKB-569, PKI-166, GW-572016, Ionafarnib, BMS-214662, tipifarnib; amifostine, NVP- 2023248067

LAQ824, suberoyl analide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951 , aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gleevac, gemcitabine, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6- mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291 , squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox,gefitinib, bortezimib, paclitaxel, cremophor-free paclitaxel, docetaxel, epithilone B, BMS- 247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA- 923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR- 3339, ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001 , ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11 , dexrazoxane, alemtuzumab, all-transretinoic

acid, ketoconazole, interleukin-2, megestrol, immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, an NK-1 receptor antagonists, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, 2023248067

pegfilgrastim, erythropoietin, epoetin alfa, darbepoetin alfa and mixtures thereof.

22. A method for inducing degradation of a target protein in a cell, the method comprising administering an effective amount of a compound of claim 1 to the cell, wherein the compound effectuates degradation of the target protein.

23. A method for treating cancer, said method comprising administering the compound of claim 1 to a patient in need thereof, wherein the compound effectuates for the treatment or alleviation of at least one symptom of cancer in the patient.

24. The method of claim 23, wherein the cancer is squamous-cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; multiple myeloma, sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor or teratocarcinomas, T-lineage Acute lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B

Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosome positive ALL and Philadelphia chromosome positive CML. 2023248067