TWI843372B - Compounds for mutant kras protein degradation and uses thereof - Google Patents

Abstract

The present disclosure provides a bifunctional compound of formula (I): RB─Linker─ULM (I), which binds more preferentially to mutant KRAS proteins than to wild KRAS protein and promotes degradation of KRAS protein via recruitment of an E3 ubiquitin ligase. Pharmaceutical compositions comprising the bifunctional compound and uses thereof are also provided.

Description

Compounds for degrading mutant KRAS protein and their applications

The present invention generally relates to bifunctional compounds that preferentially bind to mutant KRAS proteins compared to wild-type KRAS proteins and promote KRAS protein degradation by recruiting E3 ubiquitin ligases, and the use of such compounds for treating diseases associated with KRAS mutations.

The proteolysis targeting chimera (PROTAC) technology was first described in 2001 (Sakamoto et al., "Protacs: chimeric molecules that target proteins to the Skp1-Cullin-F box complex for ubiquitination and degradation", Proceedings of the National Academy of Sciences of the United States of America . 98 (15): 8554-9). PROTAC is a dual-headed heterobifunctional molecule that can remove unwanted proteins by inducing selective intracellular proteolysis. PROTAC is composed of two protein binding parts: one for binding to the E3 ubiquitin ligase and the other for binding to the target protein. PROTAC binds to two proteins and brings the target protein to the E3 ligase, causing ubiquitination of the target protein for subsequent degradation by the proteasome (Bondeson et al., "Lessons in PROTAC design from selective degradation with a promiscuous warhead." Cell Chem Biol. 25(1): 78-87, 2018). PROTAC technology has been used for several targets: AR, ER, STAT3, BTK, FLT-3, EGFR, BCR-ABL, BET, BRD7/9, CDK4/6, CK2, ALK, PI3K, MCL-1, PARP1 and c-MET.

RAS proteins are proto-oncogenes and are encoded by three RAS genes: HRAS , Kirsten ras sarcoma protein (KRas or KRAS) , and NRAS . RAS proteins function as binary switches in signaling pathways that control cell growth and differentiation, converting the inactive GDP-bound state to the active GTP-bound state. Mutations usually occur at codons 12, 13, and 61, which cause a reduction in the intrinsic GTPase activity of RAS proteins, or prevent GAP binding, activating downstream signaling pathways and contributing to tumor formation and maintenance. (Chang et al., “Detection of N-, H-, and KRAS codons 12, 13, and 61 mutations with universal RAS primer multiplex PCR and N-, H-, and KRAS-specific primer extension” Clin Biochem. 43(3):296-301, 2010.) RAS gene mutations are commonly found in a variety of malignant tumors, including pancreatic cancer (90%), colorectal cancer (45%), and lung cancer (35%). In cell and animal models, many tumor types have been shown to be associated with the persistent expression of oncogenic mutant RAS. RAS is difficult to target pharmacologically due to its picomolar affinity for GTP and GDP, and because it lacks a well-defined pocket for high-affinity small molecule binding, it is often referred to as "undruggable" (Cox et al., "Drugging the undruggable Ras: mission possible?" Nat Rev Drug Discov. 13(11): 828-851, 2014).

Currently, AMG510 (Sotorasib), a small molecule inhibitor targeting the KRAS ^G12C mutant, has passed clinical trials and was approved for marketing by the US FDA on May 28, 2021. The strategy of targeting mutant KRAS protein has been shown to be effective in treatment.

Michael J. Bond et al., "Targeted Degradation of Oncogenic KRAS ^G12C by VHL-Recruiting PROTACs" ACS Cent. Sci. 2020, 6, 8, 1367-1375; US 2019/0315732 A1; WO2019195609A2 and WO2021/207172 A1 disclose the use of KRAS ^G12C inhibitors as binders to produce degraders, thereby providing a different therapeutic strategy from inhibitors. However, both KRAS ^G12C inhibitors and KRAS ^G12C degraders are only effective against KRAS ^G12C , and are ineffective against other variants such as G12D, G12V, G13D, G12R, G12S and Q61H.

There is a continuing need for effective treatments for KRAS-related diseases and conditions, such as pancreatic cancer, colorectal cancer, lung cancer and non-small cell lung cancer, biliary malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia and breast cancer.

The present invention describes heterobifunctional compounds for recruiting KRAS (such as mutant or gain-of-function KRAS) to E3 ubiquitin ligases for targeted ubiquitination and subsequent proteasomal degradation, and methods of making and using the same. In addition, the present specification provides methods of using an effective amount of the bifunctional compounds of the present invention to treat or ameliorate disease conditions, such as KRAS-related diseases or disorders, such as accumulation or overactivity of KRAS protein or mutant or gain-of-function KRAS protein or misfolded KRAS protein, pancreatic cancer, colorectal cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia, and breast cancer.

In one embodiment, the present invention describes a bifunctional compound of formula (I): RB─Linker─ULM (I) or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, hydrate, polymorph, isotopically enriched derivative or prodrug thereof, wherein: ULM is an E3 ubiquitin ligase binding portion or a chaperone protein complex binding portion; The linker is a group covalently bonded to the RB and ULM portions; and RB is a RAS protein binding portion and is represented by the formula RB-I: wherein: _X1 is H, ^NRX1RX2 , ^SO2NRX1RX2 , _ORX1 , ^CHRX1RX2 , NH(C=O) ^RX3 or CN; ^RX1 and ^RX2 are each independently H, OH or a linear or branched _C1-4 alkyl group, which ^is optionally substituted by ^one or ^more halogen groups ^{, or RX1 and RX2} together with the atoms to which they are attached form a 4- to 8-membered ring system containing 0 to 2 heteroatoms and which is optionally substituted by one or more _C1-4 alkyl groups; ^RX3 is a linear or branched C1-10 alkyl group, which is optionally substituted by one or more halogen groups, or a linear or branched _C1-10 alkyl group, which is optionally substituted by one or more halogen groups; _W1 is a bond, _C1-6 alkyl, alicyclic, heterocyclic, bicyclic or biheterocyclic, each of which is optionally substituted by one, _two or three _RW1 , and each ^RW1 is independently H, halogen, OH, _NH2 , _NMe2 , _NEt2 , CN, _C1-4 alkyl optionally substituted by one or more F, or _C1-3 alkoxy optionally substituted by one or more F; ^R1 is CH, C _- Me, C _- halogen or N; is an aryl, heteroaryl or indazole, which is independently substituted by one or more halogen groups, CF ₃ , OCF ₃ , OR ^A1 -R ^A2 , hydroxyl, nitro, CN, C≡CH, a straight or branched C _1-6 alkyl group optionally substituted by one or more halogen groups or C _{1-6 alkoxy groups, a straight or branched C 1-6 alkoxy} group optionally substituted by one or more halogen groups, a C _2-6 alkenyl group, a C _2-6 alkynyl group or a 4- to 7-membered saturated or partially unsaturated heterocyclic ring containing 1-2 heteroatoms, ^RA1 is a C _1-6 alkyl group, ^RA2 is a heteroaryl group containing 1 to 2 heteroatoms and optionally substituted by one or more halogen groups; X ₂ is CH ₂ or C═O; is a 4- to 8-membered aliphatic ring system having 0 to 4 heteroatoms, which is optionally substituted by 0 to 4 R ^Q , each R ^Q is independently OH, or a straight or branched C _1-6 alkyl group which is optionally substituted by one or more halogen groups, or two R ^Q groups together with the atoms to which they are attached form a 3- to 8-membered ring system containing 0 to 2 heteroatoms; Z ₁ is H, C _1-4 alkyl, C _1-4 alkoxy, halogen, OH, CN or C _2-4 alkynyl; and the dotted line indicates the point of attachment to the linker.

The bifunctional compounds of formula (I) can induce ubiquitination of KRAS protein and promote its degradation in cells. The advantage of the bifunctional compounds of formula (I) provided herein is that they can have a wide range of pharmacological activities.

In another aspect, the present invention provides a pharmaceutical composition comprising an effective amount of a bifunctional compound as described herein and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides a pharmaceutical composition for preventing, ameliorating and/or treating a disease associated with KRAS mutation in a subject in need thereof, comprising an effective amount of a bifunctional compound and one or more pharmaceutically acceptable excipients.

Definition

In order to make the present invention fully understood, the following embodiments are described. The following terms are used in the specification:

It must be noted that, as used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly requires otherwise. Thus, unless the context requires otherwise, singular terms shall include pluralities and plural terms shall include the singular.

Typically, ranges are expressed herein as from "about" one particular value and/or to "about" another particular value. When such ranges are expressed, an embodiment includes a range from one particular value and/or to another particular value. Similarly, when values are expressed as approximations by using the word "about," it is understood that the particular value forms another embodiment. It should be further understood that each of the endpoints in a range has meaning both in relation to the other endpoint and independently of the other endpoint. As used herein, the term "about" refers to ±20%, preferably ±10%, and even more preferably ±5%.

The term "and/or" is used to refer to two items or either of the two items mentioned.

The terms "treatment," "treating," and "treat" generally refer to obtaining a desired pharmacological and/or physiological effect. The effect may be preventive, in terms of completely or partially preventing a disease, disorder, or symptoms thereof, and therapeutic, in terms of partially or completely curing a disease, disorder, and/or symptoms attributable thereto. "Treatment," as used herein, encompasses any treatment of a disease in a mammal, preferably a human, and includes (1) inhibiting the development of a disease, disorder, or symptoms thereof in a subject, or (2) alleviating or ameliorating a disease, disorder, or symptoms thereof in a subject.

The terms "preventing" or "prevention" are recognized in the art and when used in conjunction with a condition, include the administration of an agent prior to the onset of the condition that reduces the frequency or severity of symptoms of the medical condition or delays the onset of the condition in a subject relative to a subject who has not received the agent.

The terms "individual," "subject," and "patient" are used interchangeably herein and refer to any mammalian subject in need of diagnosis, treatment, or therapy.

The term "effective amount" of the active ingredient provided herein means a sufficient amount of the desired regulated ingredient to provide the desired function. As will be noted below, the exact amount required will vary from individual to individual, depending on the individual's disease state, physical condition, age, sex, species and weight, the specific characteristics and formulation of the composition, etc. The dosing regimen can be adjusted to induce the optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the urgency of the therapeutic situation. Therefore, it is impossible to specify an exact "effective amount". However, a person generally skilled in the art can determine the appropriate effective amount using only routine experiments.

Unless otherwise specified, as used herein, the term "alkyl" refers to a monovalent saturated straight or branched chain alkyl group containing 1 to 12 carbon atoms. Preferably, the alkyl group is a C ₁ -C ₈ alkyl group. More preferably, the alkyl group is a C ₁ -C ₆ alkyl group. The alkyl group may be substituted or unsubstituted. Examples of C ₁ -C ₆ alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, dibutyl, tertiary butyl, pentyl (including all isomeric forms), hexyl (including all isomeric forms), heptyl (including all isomeric forms), and octyl (including all isomeric forms).

Heteroatoms such as oxygen, sulfur, and nitrogen (in the form of a tertiary amine moiety) may be present in an alkylene group to yield _a "heteroalkylene." Examples of _{heteroalkylene} groups include, but are _not limited to _{, -CH2CH2N} ( _CH3 ) ₂ and _{-CH2CH2OCH2CH3} .

Unless otherwise specified, as used herein, the term "alkoxy" refers to a radical of the formula -O-(alkyl), wherein alkyl is as defined above. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, di-butoxy, tertiary butoxy, n-pentoxy, and n-hexoxy.

As used herein, the term "aryl" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π electron system. Aryl groups may have 6 to 14 carbon atoms in the ring. Exemplary aryl groups include, but are not limited to, phenyl, biphenyl, and naphthyl.

As used herein, the term "cycloalkyl" refers to an all-carbon monocyclic or fused-ring (i.e., rings that share a pair of adjacent carbon atoms) radical, wherein one or more of the rings do not have a completely conjugated π electron system. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, 1-methylcyclopropyl, 2-methylcyclopentyl, and 2-methylcyclooctyl.

As used herein, the terms "heterocycle" and "heterocyclic group" are used interchangeably. The term "heterocycle" or "heterocyclic group" refers to a monocyclic, bicyclic or polycyclic structure having 3 to 14 atoms, or 3 to 12 atoms, or 3 to 10 atoms, or 3 to 8 atoms, or 4 to 7 atoms, or 5 or 6 atoms; wherein one or more atoms, such as 1, 2 or 3 atoms, are independently selected from the group consisting of N, O and S, and the remaining ring-forming atoms are carbon atoms. The ring structure may be saturated or unsaturated, but is not aromatic. Exemplary heterocyclic rings include, but are not limited to, imidazolyl, imidazolinyl, imidazolidinyl, quinolinyl, isoquinolinyl, indolyl, indazolyl, indazolinyl, perhydropyrimidine, pyridazyl, pyridinyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrimidine, quinoxalinyl, piperidinyl, pyranyl, pyrazolinyl, piperimidine, pyridazinyl, thiophene, thiophene, furanyl, thiophenyl, triazolyl, thiazolyl, carbolinyl, tetrazolyl, thiazolidinyl, benzofuranyl, thiophene, thiophene, thiazolyl, thiazolyl, benzofuranyl ... , oxazolyl, benzoxazolyl, oxopiperidinyl, oxopyrrolidinyl, oxazolidinyl, azolide, isoxazolyl, isothiazolyl, furazolyl, tetrahydropyranyl, tetrahydrofuranyl, thiadiazolyl, dioxacyclopentenyl, dioxacyclohexenyl, oxathiacyclopentenyl , benzodioxacyclopentyl, dithiocyclopentyl, thienyl, tetrahydrothienyl, cyclobutanesulfonyl, dioxacyclohexanyl, dioxolane, tetrahydrofurodihydrofuranyl, tetrahydropyranodihydrofuranyl, dihydropyranyl, tetrahydrofurofuranyl and tetrahydropyranofuranyl.

As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic or tricyclic aromatic group containing 1 to 4 heteroatoms selected from S, N and O, and includes a group having two such monocyclic rings or one such monocyclic ring and a monocyclic aromatic ring directly linked by a covalent bond. The heteroaryl group may have 5 to 14 ring-forming atoms, including 1 to 13 ring-forming carbon atoms and 1 to 8 ring-forming heteroatoms each independently selected from O, S and N. Exemplary heteroaryl groups include, but are not limited to, thienyl, benzothienyl, furanyl, benzofuranyl, pyrrolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, benzoisothiazolyl, pyrazolyl, oxazolyl, benzooxazolyl, isoxazolyl, benzoisooxazolyl, isothiazolyl, triazolyl, benzotriazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyrimidinyl, pyrimidinyl, tririmidinyl, indolyl, and indazolyl.

As used herein, the term "heterocycloalkyl" refers to a monocyclic or polycyclic (including 2 or more rings fused together, including spiro, fused or bridged systems, such as bicyclic systems) saturated or unsaturated non-aromatic 4-15 membered ring system, including 1 to 14 ring-forming carbon atoms and 1 to 10 ring-forming heteroatoms independently selected from O, S and N. Examples of heterocycloalkyl include, but are not limited to, azacyclobutyl, tetrahydrofuran, dihydrofuran, dioxane, oxoquinoline, etc.

As used herein, the term "ring system" refers to one or more rings (e.g., monocyclic or polycyclic) of atoms, which may be saturated, unsaturated or aromatic, and the atoms may be only carbon or may include heteroatoms (e.g., N, S, O). For example, the ring system may include, but is not limited to, aryl (e.g., phenyl, heterophenyl, naphthyl), alicyclic, heterocyclic, bicyclic (including spiro, fused and bridged) and biheterocyclic structures.

As used herein, the terms "halide" and "halo" are used interchangeably and include fluorine, chlorine, bromine and iodine.

Compound

In one embodiment, the present invention provides a heterobifunctional compound comprising an E3 ubiquitin ligase binding moiety ("ULM") covalently coupled to a protein targeting moiety (RB) via a chemical bonding group (linker) according to structure (I): RB─Linker─ULM     (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, tautomer, stereoisomer, isotopically enriched derivative or prodrug thereof, wherein: The linker is a group covalently bonded to the RB and ULM moieties, RB is a RAS protein binding moiety that binds to a mutant or wild-type KRAS protein; and ULM includes all E3 ubiquitin ligase binding moieties or chaperone complex binding moieties.

The bifunctional compounds described herein preferentially interact with KRAS mutant proteins compared to KRAS wild-type proteins. As described herein, the therapeutic effect may be the result of the compounds described herein degrading, regulating, binding or modifying KRAS proteins. Without wishing to be bound by any particular theory, the therapeutic effect may be the result of the compounds described herein regulating, targeting, binding or modifying E3 ubiquitin ligases. The therapeutic effect may be the result of recruiting E3 ubiquitin ligases to ubiquitinated RAS proteins and marking them for proteasomal degradation by regulating, targeting, binding or modifying E3 ubiquitin ligases by compounds. Specifically, the bifunctional compounds of formula (I) are suitable for treating and/or preventing RAS mutation-related diseases in individuals in need thereof.

ULM

The ULM is an E3 ubiquitin ligase binding portion or a chaperone protein complex binding portion, and can be a Cereblon E3 ubiquitin ligase binding portion (CLM), a Von Hippel-Lindau (VHL) E3 ubiquitin ligase binding portion (VLM), a DDB1-related and CUL4-related factor 16 (DCAF16) E3 ubiquitin ligase binding portion (DLM), an IAP E3 ubiquitin ligase binding portion (ILM), a mouse double microsome 2 (MDM2) homolog E3 ubiquitin ligase binding portion (MLM), a Kelch-like ECH-associated protein-1 (KEAP1) E3 ubiquitin ligase binding portion, a DCAF15 E3 ubiquitin ligase binding portion, a RNF4 E3 ubiquitin ligase binding portion, a RNF114 E3 ubiquitin ligase binding portion, aromatic hydrocarbon receptor (AhR) E3 ubiquitin ligase binding portion, or other E3 ubiquitin ligase binding portion described in SLAS Discovery, 1-19, 2020. ULM includes all parts that bind, can bind, or form a covalent bond with any E3 ubiquitin ligase. For example, in some embodiments, ULM is capable of binding to E3 ubiquitin ligases such as celebron or van Heppel-Lindau (VHL). In some embodiments, ULM is capable of forming a covalent bond with an E3 ubiquitin ligase such as DCAF16. In some embodiments, ULM is capable of binding to a variety of different E3 ubiquitin ligases. In some embodiments, ULM is bound to celebron. In some embodiments, ULM is bound to VHL. In certain embodiments, ULM forms a covalent bond with DCAF16.

Serebron is an E3 ubiquitin ligase, and it forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), tardatin-4A (CUL4A), and regulatory factor of tardatin 1 (ROC1). This complex ubiquitinates a variety of proteins.

In certain embodiments, the ULM is a celebron E3 ligase binding moiety (CLM) selected from the group consisting of thalidomide, lenalidomide, pomalidomide, analogs thereof, isosteres thereof, or derivatives thereof; preferably having the following formulas CLM-a and CLM-b: CLM-a wherein: W is selected from CH ₂ and C=O; and Q ₁ , Q ₂ , Q ₃ and Q ₄ are each independently C, N or C-halogen, and wherein one of Q ₁ , Q ₂ , Q ₃ and Q ₄ is covalently linked to a linker; CLM-b where the dashed lines indicate the connection points with the connectors.

In some embodiments, the ULM is ; wherein the dotted line indicates the connection point with the linker; and X _CLM is a halogen.

Van Hippel-Lindau (VHL) is an E3 ubiquitin ligase. VHL comprises a substrate recognition subunit/E3 ubiquitin ligase complex VCB, which includes elongins B and C, and a complex including tachylin-2 and Rbxl. The major substrates of VHL are hypoxia-induced factor 1 (HIF-1a), a transcription factor that upregulates genes in response to low oxygen levels, such as the angiogenic growth factor VEGF, and the red blood cell inducing interleukin, erythropoietin. VCB is a known target in cancer, chronic anemia, and ischemia. In one embodiment, ULM is a VHL E3 ubiquitin ligase binding moiety and may be hydroxyproline or a derivative thereof.

In certain embodiments, the ULM comprises a peptide backbone structure. In certain embodiments, the ULM has a chemical structure represented by the following formula: wherein: R ₅ is H or a linear or branched C _1-3 alkyl group; R ₆ is CN or a 5-membered heteroaryl group having one or two heteroatoms selected from N, S or O, which is optionally substituted with a methyl group (e.g., , the dotted line indicates the point of connection with the benzene ring); Z ₂ is F or CN; and the dotted line indicates the point of connection with the linker.

As described in Xiaoyu Zhang et al., Nature Chemical Biology , Vol. 15, 2019, p73791og, DDB1-associated and CUL4-associated factor 16 (DCAF16) is a substrate-recognized component of the CUL4-DDB1 E3 ubiquitin ligase that lacks a characteristic feature. The DCAF16 protein has eight cysteine residues and can covalently bind to the DCAF16 binding portion of the heterobifunctional degrader at the cysteine residues and subsequently promote protein degradation.

In certain embodiments, the ULM is a DCAF16 ligase binding moiety (DLM) and has the chemical structure represented by D16-a: D16-a wherein: R ₇ is H, halogen, C _1-4 alkyl, C _1-4 alkoxy; Y ₁ is O or S or NH; R ^m is a covalent electrophile and is selected from the following groups: , the dashed line indicates the point of attachment to the nitrogen atom; and the dashed line to Y indicates the point of attachment to the linker.

In some embodiments, the ULM is a DLM represented by: ; The dotted line represents the connection point with the connector.

In certain embodiments, the ULM binds to an E3 ubiquitin ligase with a _KD value of less than about 10,000 nM, less than about 5,000 nM, less than about 1,000 nM, less than about 500 nM, less than about 100 nM, or less than about 50 nM.

In certain embodiments, the ULM binds celebron with a _KD value of less than about 50 μM, less than about 10,000 nM, less than about 5,000 nM, less than about 1,000 nM, less than about 500 nM, less than about 100 nM, less than about 50 nM.

In certain embodiments, the ULM binds VHL with a _KD value of less than about 50 μM, less than about 10,000 nM, less than about 5,000 nM, less than about 1,000 nM, less than about 500 nM, less than about 100 nM, less than about 50 nM.

RB

In certain embodiments, RB is represented by the formula RB-I: , wherein: _X1 is H, ^NRX1RX2 , ^{SO2NRX1RX2 ,} _ORX1 , ^CHRX1RX2 , NH(C= ^O ) ^RX3 or CN; ^RX1 and ^RX2 are each independently H, OH or _C1-4 alkyl ⁽ straight chain or branched chain, optionally substituted by 1 or more halogen groups), or ^RX1 and ^RX2 together with the ^atoms to which they are attached form a 4- to 8-membered ring system containing 0 to 2 heteroatoms (optionally substituted by one or more _C1-4 alkyl groups); ^RX3 is _C1-10 alkyl (straight chain or branched chain, optionally substituted by 1 or more halogen groups), C W1 is a bond, C1-6 alkyl, alicyclic, heterocyclic, bicyclic or biheterocyclic _{, each of which is optionally substituted by 1, 2 or 3 RW1; and each RW1 is independently halogen, C1-4 alkyl (optionally substituted by 1 or more F), C1-3} ^alkoxy ₍ ^optionally _{substituted by 1 or} more F), OH _{, NH2} , _NMe2 , _NEt2 or CN; R ₁ is CH, C-Me, C-halogen or N; is an aryl or heteroaryl group, which is independently substituted by: 1 or more OR ^A1 -R ^A2 , halogen, CF ₃ , OCF ₃ , hydroxyl, nitro, CN, C≡CH, C _1-6 alkyl (straight chain or branched chain, optionally substituted by 1 or more halogen, C _1-6 alkoxy), C _1-6 alkoxy (straight chain or branched chain, optionally substituted by 1 or more halogen), C _2-6 alkenyl, C _2-6 alkynyl, or a 4- to 7-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, R ^A1 is a C _1-6 alkyl group, R ^A2 is a heteroaryl group containing 1 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and optionally substituted by 1 or more halogen groups; _X2 is _CH2 or C=O; is a 4- to 8-membered alicyclic ring having 0 to 4 heteroatoms, which is optionally substituted by 0 to 4 R ^Q , each R ^Q is independently OH, or C _1-6 alkyl (straight or branched, optionally substituted by 1 or more halogen groups), or 2 R ^Q groups together with the atoms to which they are attached form a 3- to 8-membered ring system containing 0 to 2 heteroatoms; Z ₁ is H, C _1-4 alkyl, C _1-4 alkoxy, halogen, OH, CN or C _2-4 alkynyl; and the dotted line indicates the point of attachment to the linker.

In certain embodiments, the impurity atoms in ^RX1 and ^RX3 are independently selected from nitrogen, oxygen and sulfur. Preferably, the impurity atom is nitrogen.

In certain embodiments, _X1 is H, ^NRX1RX2 , ^SO2NRX1RX2 , ^ORX1 , ^CHRX1RX2 , NH(C=O) ^RX3 or CN; ^RX1 and ^RX2 are each independently H or _C1-4 alkyl, or ^RX1 and ^RX2 together ^with the atoms to which ^they are attached form a 4- to 8-membered aryl, alicyclic or bicyclic ring containing 0 to 2 nitrogen atoms and optionally substituted with 1 ^{or more C1-4 alkyl groups; and RX3} _{is C1-10 alkyl} ⁽ _straight or branched, optionally substituted with 1 or more halogen groups), C _1-10 alkoxy (straight or branched chain, optionally substituted by 1 or more halogen groups) or aryl or alicyclic (4- to 8-membered ring system containing 0 to 2 heteroatoms independently selected from nitrogen and oxygen, optionally substituted by 1 or more halogen groups, CF ₃ or NH ₂ ).

In certain embodiments, W ₁ is a bond or a C _1-4 alkyl group which is optionally substituted with a C _1-3 alkoxy group.

In some embodiments, is phenyl, heterophenyl, naphthyl or indazole, which is independently substituted by 1 or more O- _CH2 - ^RA2 , halogen, _CF3 , _OCF3 , hydroxyl, CN, _C1-6 alkyl, _C1-6 alkoxy, and ^RA2 is a heteroaryl group having 1 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur and optionally substituted by 1 or more halogen groups.

In some embodiments, is phenyl or indazole, which is independently substituted with 1 or more O- _CH2 - ^RA2 , halogen, _CF3 , _OCF3 , CN, _C1-6 alkyl, _C1-6 alkoxy, ^RA2 is a heteroaryl group having 1 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and optionally substituted with 1 or more halogen.

In some embodiments, The impurity atom is independently selected from nitrogen, oxygen and sulfur. Preferably, the impurity atom is nitrogen.

In some embodiments, It is a 4- to 6-membered alicyclic ring having 1 to 2 heteroatoms, which is optionally substituted by 0 to 4 R ^Q groups, each R ^Q being independently H or C _1-6 alkyl (straight chain or branched chain), or 2 R ^Q groups together with the atoms to which they are attached form a 4- to 5-membered ring system containing 0 to 2 heteroatoms, and the heteroatom is nitrogen.

In certain embodiments, Formula RB-I is selected from the group consisting of RB-1 to RB-7: . The dotted lines represent the connection points with the connectors.

In certain embodiments, Formula RB-I is represented by: ; The dotted line represents the connection point with the connector.

In some embodiments of the present invention, RB-I is selected from the group consisting of: ; The dotted line represents the connection point with the connector.

In some embodiments of the present invention, the bifunctional compound is selected from Formula Ia to Formula Ig: .

In certain embodiments, the RB binding moiety binds to a KRAS protein with a KD value of less than about 100 μM, less than about 50 μM, less than about 10 μM, less than about 5 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM _, or less than about 50 nM.

In certain embodiments, the RB binding moiety selectively binds to a mutant KRAS protein relative to a wild-type KRAS protein. In certain embodiments, the compound of Formula (I) selectively binds to a mutant KRAS protein relative to a wild-type KRAS protein. In certain embodiments, the selectivity is between about 2-fold and about 5-fold. In certain embodiments, the selectivity is between about 5-fold and about 10-fold. In certain embodiments, the selectivity is between about 10-fold and about 20-fold.

Connector

The linker is a divalent moiety that connects RB and ULM; preferably, the linker covalently couples RB to ULM. In certain embodiments, the linker comprises the following chemical structure , wherein: the dotted line of the linker is the point of connection to RB or ULM; Y ^L2 is a bond, or an unsubstituted or substituted straight or branched C _1-4 alkyl or alkoxy group (e.g., optionally substituted with halogen, C _1-3 alkyl, methyl or ethyl); W ^L3 is a bond, -C=OCH ₂ -, or a 3- to 7-membered ring system (e.g., a 4- to 6-membered cycloalkyl, heterocycloalkyl or aryl) or an 8- to 12-membered spiro ring, each having 0 to 4 heteroatoms (e.g., 0 to 4 heteroatoms independently selected from N, O and S) and optionally substituted with halogen or methyl; Y ^L3 is a bond or a C _1-32 alkylalkenyl or alkynyl group, wherein one or more C atoms are optionally substituted with O, or NH 3, and each carbon is optionally substituted by halogen, =0, methyl or ethyl, and each nitrogen is optionally substituted by methyl or ethyl; Y ^L4 is a bond, O or unsubstituted or substituted straight or branched C _1-6 alkyl, one or more of which is optionally replaced by O, NH or NCH ₃ , and optionally substituted by halogen or methyl; W ^L4 is a 3- to 8-membered ring system (e.g., a 4- to 6-membered cycloalkyl or heterocycloalkyl, aryl or ) or a 5- to 8-membered spiro ring, each having 0 to 4 heteroatoms (e.g., 0 to 4 heteroatoms independently selected from N, O and S) and optionally substituted with a halogen (e.g., F, Cl, Br) or a methyl group; and Y ^L5 is a bond or an unsubstituted or substituted C _1-6 alkyl group, wherein one or more C atoms are optionally replaced with O and optionally substituted with a halogen (e.g., F, Cl, Br) or a methyl group.

In certain embodiments, Y ^L2 is a bond or C _1-4 alkyl; preferably a bond or CH ₂ .

In certain embodiments, W ^L3 is a bond, -C=OCH ₂ -, a 4- to 6-membered heterocycloalkyl containing 0-2 nitrogen atoms, or an 8- to 12-membered spiro ring, each ring containing 0-1 nitrogen atoms.

In certain embodiments, Y ^L3 is a bond or a C _1-8 alkyl or alkynyl group, wherein one or more C atoms are optionally replaced by O and each carbon is optionally substituted by =0.

In certain embodiments, Y ^L4 is a bond or C _1-4 alkyl; preferably a bond or CH ₂ .

In certain embodiments, W ^L4 is aryl which is optionally substituted with a halogen (eg, F, Cl, Br) or a 4- to 6-membered heterocycloalkyl group containing 0 to 2 nitrogen atoms.

In certain embodiments, Y ^L5 is a bond or C _1-4 alkyl; preferably a bond or CH ₂ .

In some embodiments, the connector is selected from the group consisting of: ; The dotted line of the connector is the connection point with RB or ULM.

In certain embodiments, the ring system of the linker and the ring system of RB may share one or more atoms (carbon atoms or heteroatoms, preferably carbon atoms) to form a bicyclic structure. For example, the ring system of the linker and the ring system of RB may share one atom to form a spirocyclic structure, share two atoms to form a fused bicyclic structure, or share three atoms to form a bridged bicyclic structure.

The RB group and the ULM group can be covalently linked to the linker via any group that is suitable and stable to the chemical properties of the linker. In exemplary aspects of the present invention, the linker is independently covalently linked to the RB group and the ULM group via amide, ester, thioester, keto, carbamate (carbamate), carbon or ether in certain embodiments, each of which can be inserted at any position on the RB group and the ULM group to maximize the binding of the ULM group on the ubiquitin ligase to the RB group on the target protein to be degraded.

WO 2021/222138 also discloses bifunctional compounds that bind to mutant KRAS proteins. In certain embodiments of the present invention, although not wishing to be bound by any particular theory, it is believed that due to the linker being linked to RB-I , and/or because the molecular weight of the bifunctional compounds is relatively small (smaller than the molecular weight of WO 2021/222138), the bifunctional compounds provide improved pharmacokinetic properties and improved therapeutic effects.

As used herein, the phrase "substituted or unsubstituted" means that substitution is optional. Where substitution is desired, such substitution means that any number of hydrogens on the designated atom are replaced by a selected group selected from the designated group, provided that the normal valence of the designated atom is not exceeded and that the substitution produces a stable compound. For example, when the substituent is keto (i.e., =0), then 2 hydrogens on the atom are replaced. Examples of substituents for "substituted" groups are substituents found in the exemplary compounds and examples disclosed herein, and may include, for example, halogen, -OH, _-CF3 , -CN, _-NO2 , alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, haloalkyl, alkylamino, aminoalkyl, dialkylamino, hydroxyalkyl, alkoxyalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, alkylaminoalkoxy, alkylaminoalkyl, and aryl, and the like.

As used herein, the term "pharmaceutically acceptable salt" refers to the compounds according to the present invention used in the form of salts derived from inorganic or organic acids and bases. Acid salts include, for example, the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, hydrogen sulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, fluoroheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, Salts derived from suitable bases include alkali metals (e.g., sodium), alkali earth metals (e.g., magnesium), ammonium, and NW ⁴⁺ (wherein W is a C _1-4 alkyl group).

As used herein, "prodrugs" are intended to include any covalently bonded carriers that, when such prodrugs are administered to a subject, release the active parent drug according to formula (I) through physiological actions in vivo (such as hydrolysis, metabolism and the like). Those of ordinary skill in the art are familiar with the applicability and techniques involved in preparing and using prodrugs. Prodrugs of compounds of formula (I) (parent compounds) can be prepared by modifying functional groups present in the compound so that the modifications are cleaved to the parent compound in conventional manipulations or in vivo. "Prodrugs" include compounds of formula (I) in which a hydroxyl, amine or sulfhydryl group is bonded to any group that, when the prodrug is administered to a subject, is cleaved to form a free hydroxyl, free amine or free sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, derivatives and metabolites of compounds of formula (I) that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureas, and biohydrolyzable phosphate analogs. In certain embodiments, the prodrug of a compound of formula (I) having a carboxyl functional group is a lower alkyl (e.g., C ₁ -C ₆ ) ester of a carboxylic acid. Carboxylic acid esters are preferably formed by esterifying any carboxylic acid moiety present on the molecule.

The compounds of the present invention may exist in the form of a solvent complex. As used herein and unless otherwise indicated, the term "solvent complex" means a compound of formula (I) or a pharmaceutically acceptable salt thereof further including a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. If the solvent is water, the solvent complex may be appropriately referred to as a "hydrate", such as a hemihydrate, a monohydrate, a sesquihydrate, a dihydrate, a trihydrate, etc.

As used herein, the term "tautomer" refers to compounds whose structures differ significantly in atomic arrangement, but which readily and rapidly reach equilibrium, and it is understood that the compounds provided herein may be depicted as different tautomers, and when compounds have tautomeric forms, all tautomeric forms are intended to be within the scope of the present invention, and the naming of the compounds does not exclude any tautomers. Exemplary tautomerizations include, but are not limited to, amide-imide; enamine-imine; enamine-(different)enamine tautomerizations; and keto-enol.

The term "stereoisomers" refers to compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space. Stereoisomers include non-mirror isomers, mirror isomers, conformational isomers and the like.

The term "polymorph" refers to a crystalline form of a compound (or its salts, hydrates, or solvates). All polymorphs have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability, and solubility. The recrystallization solvent, crystallization rate, storage temperature, and other factors can cause one crystalline form to predominate. Various polymorphs of a compound can be prepared by crystallization under different conditions.

As used herein, an "isotopically enriched derivative" refers to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom. "Isotopic enrichment" may be expressed in terms of the percentage of incorporation of a certain amount of a particular isotope at a specified atom in a molecule, rather than in terms of the natural isotopic abundance of that atom.

The bifunctional compounds of the present invention may exist in one or more specific geometric, optical, mirror isomers, non-mirror isomers, epimers, hysteresis isomers, stereoisomers, tautomers, configurational isomers or rotamer forms, including but not limited to cis and trans; E- and Z-forms; c-, t- and r-forms; endo- and exo-forms; R-, S- and meso-forms; D- and L-forms; d- and I-forms; (+) and (-) forms; keto-, enol- and enolate-forms; cis- and trans-forms; syncline- and anticline-forms; α- and β-forms; axial and equatorial forms; boat-, chair-, torsion-, envelope- and semi-chair-forms; and combinations thereof.

In certain embodiments, the bifunctional compound is selected from compound (cpd) 1-72: 4-amino-N-(3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidin-4-carboxamide (cpd 1); 4-amino-N-(3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidin-4-carboxamide (cpd 2); 5-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 3); 5-(4-((4-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 4); 5-(4-((1-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 5); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 6); 5-(4-((4-((1-(3-aminopropyl)-3-(4-fluorophenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 7); 5-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 8); 1-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-((1-(2-chloroacetyl)-1,2,3,4-tetrahydroquinolin-6-yl)oxy)ethan-1-one (cpd 9); 5-(4-((4-((1-(3-aminopropyl)-3-(4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 10); 4-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-1H-indol-3-yl)benzonitrile (cpd 11); 5-(4-((1-((1-(3-aminopropyl)-3-(6-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 12); 3-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-1H-indol-3-yl)benzonitrile (cpd 13); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-7-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 14); 5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 15); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-6-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 16); 5-(4-((1-((1-(3-aminopropyl)-3-(2-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 17); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethyl)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 18); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-4-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 19); 4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 20); (3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamic acid tributyl ester (cpd 21); 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)prop-1-yn-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 22); 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)propyl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 23); 5-(4-(2-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)acetyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 24); 5-(4-((1-((1-(3-aminopropyl)-3-(3-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (cpd 25); 5-(4-((1-(1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indole-5-carbonyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (Cpd 26); 5-(4-((1-((1-(3-aminopropyl)-3-(3-hydroxynaphthalen-1-yl)-1H-indole-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxypiperidin-3-yl)isoindole-1,3-dione (Cpd 27); 5-(4-((1-((1-(Azocyclobutane-3-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 28); 5-(4-((1-((1-(3-aminopropyl)-3-(5-chloro-6-methyl-1H-indazole-4-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 29); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (Cpd 30); (2S)-N-(2-(3-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carboxamide (Cpd 31); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindole-1,3-dione (Cpd 32); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(methylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 33); 5-(7-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 34); 5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-((3-fluoropyridin-2-yl)methoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 35); 5-(4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)azocyclobutane-3-yl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 36); 5-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 37); 5-(4-((1-((1-(5-aminopentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 38); 5-(7-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 39); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)acetamide (Cpd 40); 4-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)butyronitrile (Cpd 41); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-((1-methyl-1H-imidazol-5-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 42); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-methoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 43); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(2,2,2-trifluoroethoxy)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindolin-1,3-dione (Cpd 44); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxopiperidin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-1H-pyrrole-2-carboxamide (Cpd 45); 3-((4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 46); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-3-(trifluoromethyl)benzamide (Cpd 47); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-4-fluorobenzamide (Cpd 48); 3-((4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 49); 5-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 50); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 51); 5-(4-((1-((1-((2,3-dihydrobenzo[b][1,4]dioxadien-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 52); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(pyridin-2-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 53); 3-((4-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 54); 3-((4-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 55); 5-(4-((1-((1-((1,3-dioxolan-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 56); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (Cpd 57); 3-((4-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 58); 5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidin]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (Cpd 59); 3-((4-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 60); 4-amino-N-(3-(5-((4-((4-((2,6-bipiperidin-3-yl)amino)-2-fluorophenyl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidine-4-carboxamide (Cpd 61); 5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-indene-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-N,N-dimethyl-3-(4-(trifluoromethoxy)phenyl)-1H-indole-1-sulfonamide (Cpd 62); 3-((3-fluoro-4-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indole-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)phenyl)amino)piperidin-2,6-dione (Cpd 63); 3-((4-fluoro-3-(4-((1-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)phenyl)amino)piperidin-2,6-dione (Cpd 64); 3-((3-fluoro-4-(1'-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidin]-1-yl)phenyl)amino)piperidin-2,6-dione (Cpd 65); 5-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 66); 3-((4-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-fluorophenyl)amino)piperidin-2,6-dione (Cpd 67); 5-(4-((1-((1-(3,3-dimethoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 68); 5-(5-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 69); 5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[3,3'-azocyclobutane]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 70); 5-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidin]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 71); (2R,4S)-1-((R)-2-(2-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)acetamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((R)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Cpd 72); or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, tautomer, stereoisomer, isotopically enriched derivative or prodrug thereof.

In certain embodiments, the bifunctional compounds described herein bind to KRAS protein with a KD value of less than about 100 μM, less than about 50 μM, less than about 10 μM, less than about 5 μM, less than _about 1 μM, less than about 500 nM, less than about 100 nM, or less than about 50 nM.

In certain embodiments, the bifunctional compounds described herein selectively bind to mutant KRAS proteins relative to wild-type KRAS proteins. In certain embodiments, the compounds of formula (I) selectively bind to mutant KRAS proteins relative to wild-type KRAS proteins. In certain embodiments, the selectivity is between about 2-fold and about 5-fold. In certain embodiments, the selectivity is between about 5-fold and about 10-fold. In certain embodiments, the selectivity is between about 10-fold and about 20-fold.

In certain embodiments, the bifunctional compounds described herein bind to an E3 ubiquitin ligase with a KD value of less than about 50 μM, less than about 10,000 nM, less than about 5,000 nM, less than about 1,000 nM, less than about 500 nM, less than about 100 nM, or less than about 50 nM _.

In certain embodiments, the bifunctional compounds described herein promote degradation of mutant KRAS protein by at most 10%, at most 20%, at most 30%, at most 40%, at most 50%, at most 60%, at most 70%, at most 80%, at most 90%, at most 100% at a concentration of 20 μM or less, 10 μM or less, 5 μM or less, 1,000 nM or less, 500 nM or less, 100 nM or less, 50 nM or less, 10 nM or less.

Pharmaceutical compositions and applications

The bifunctional compounds described herein can be administered therapeutically in the form of pure chemicals, but it may be appropriate to administer the compounds in the form of pharmaceutical compositions or formulations. Therefore, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the bifunctional compounds described herein or their pharmaceutically acceptable salts, tautomers, stereoisomers, solvates, hydrates, polymorphs, isotopically enriched derivatives or prodrugs, and one or more pharmaceutically acceptable excipients.

The pharmaceutical composition can be administered in a variety of dosage forms, including but not limited to solid or liquid dosage forms, oral dosage forms, parenteral dosage forms, intranasal dosage forms, suppositories, buccal tablets, sugar-coated tablets, buccal tablets, controlled release dosage forms, pulsatile release dosage forms, rapid release dosage forms, intravenous solutions, suspensions, or combinations thereof. The pharmaceutical composition can be administered, for example, by oral or parenteral routes, including intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, respiratory (aerosol), rectal, vaginal, and topical (including intrabuccal and sublingual) administration.

"Excipients" generally refer to substances, often inert substances, added to a pharmacological composition or otherwise used as a vehicle to further facilitate administration of a compound. Examples of excipients include, but are not limited to, inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, coloring agents, preservatives, foaming mixtures, and adsorbents. Suitable inert diluents include, but are not limited to, sodium and calcium carbonates, sodium and calcium phosphates, lactose, and the like. Suitable disintegrants include, but are not limited to, starches (such as corn starch), cross-linked polyvinyl pyrrolidone, agar, alginic acid or its salts (such as sodium alginate), and the like. Binders may include, but are not limited to, magnesium aluminum silicate, starches (such as corn, wheat or rice starch), gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and the like. Lubricants, if present, will typically be magnesium and calcium stearate, stearic acid, talc or hydrogenated vegetable oils. If necessary, tablets may be coated with materials such as monostearate or distearate to delay absorption in the gastrointestinal tract. The composition may also be formulated as chewable tablets, for example by using substances such as mannitol in the formulation.

The term "therapeutically effective amount" refers to an amount of the bifunctional compound described herein alone or in combination with an anticancer agent that provides the desired effect in the treated subject after being administered to the subject in a single or multiple doses.

The bifunctional compounds described herein are effective in treating or ameliorating KRAS mutation-related diseases. When the target protein is located near the E3 ubiquitin ligase, degradation of the target protein will occur, thereby degrading the target protein/inhibiting the action of the target protein and controlling the protein level. The control of protein levels achieved by the present invention provides treatment for disease conditions or symptoms, which are regulated by the target protein by reducing the protein level in the patient's cells. "KRAS mutation-related diseases" can be cancer, autoimmune diseases, infectious diseases, or angiogenic diseases. The cancer may be lung cancer (e.g., non-small cell lung cancer), colon cancer, colorectal cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, bladder cancer, stomach cancer, kidney cancer, salivary gland cancer, ovarian cancer, uterine corpus cancer, cervical cancer, oral cancer, skin cancer, brain cancer, lymphoma, leukemia, bile duct malignancy, endometrial cancer, or myeloid leukemia.

The bifunctional compounds described herein can be administered as the sole active agent, or can be administered separately, sequentially or together with one or more other anticancer agents. Unless otherwise specified, as used herein, the term "anticancer agent" refers to an agent that can inhibit or prevent tumor growth or detect the maturation and proliferation of malignant cells (cancer cells). Anticancer agents suitable for use in combination with the compound of formula (I) include, but are not limited to, targeted cancer drugs, such as trastuzumab, ramucirumab, vismodegib, sonidegib, bevacizumab, everolimus, tamoxifen, toremifene, fulvestrant, anastrozole, exemestane, lapatinib, letrozole, pertuzumab, ado-trastuzumab emtansine, palbociclib, cetuximab, panitumumab, ziv-aflibercept, regorafenib, lmatinib mesylate, lanreotide acetate, sunitinib, denosumab, alitretinoin, sorafenib, pazopanib, temsirolimus, everolimus, tretinoin, dasatinib, nilotinib, bosutinib, rituximab uximab), alemtuzumab, ofatumumab, obinutuxumab, ibrutinib, idelalisib, blinatumomab, soragenib, crizotinib, erlotinib, gefitinib, afatinib dimaleate, ceritnib, ramucirumab, nivolumab, pembrolizumab, osimertinib, and necitumumab; alkylating agents such as busulfan, chlorambucil, cyclophosphamide, iphosphamide, melphalan, nitrogen mustard, streptozocin, thiotepa, uracil nitrogen mustard, mustard), triethylenemelamine, temozolomide, and 2-chloroethyl-3-sarcosinate-1-nitrosourea (SarCNU); antibiotics or plant alkaloids, such as actinomycin-D, bleomycin, cryptophycins, daunorubicin, doxorubicin, idarubicin, irinotecan, L-asparaginase, mitomycin-C (mitomycin-C), mitramycin, navelbine, paclitaxel, docetaxel, topotecan, vinblastine, vincristine, teniposide (VM-26), and etoposide (VP-16); hormones or steroids, such as 5α-reductase inhibitors, aminoglutethimide, anastrozole, bicalutamide, chlorotrianisene, diethylstilbestrol (DES), dromostanolone, estramustine, ethinyl estradiol (ET-16); estradiol), flutamide, fluoxymesterone, goserelin, hydroxyprogesterone, letrozole, leuprolide, medroxyprogesterone acetate, megestrol acetate, methyl prednisolone, methyltestosterone, mitotane, nilutamide, prednisolone, arzoxifene (SERM-3), tamoxifen, testolactone, testosterone, triamicnolone, and zoladex; synthetics, such as all-trans retinoic acid, carmustine (BCNU), carboplatin (CBDCA), lomustine (CCNU), cis-diaminedichloroplatinum (cis-platinum), dacarbazine, gliadel, hexamethylmelamine, hydroxyurea, levamisole, mitoxantrone, o,p'-dichlorodiphenyldichloroethane (o,p'-DDD) (also known as lysodren or mitotane), oxaliplatin, porfimer sodium, procarbazine, and imatinib mesylate (Gleevec ^® ); anti-metabolites such as chlorodeoxyadenosine, cytosine arabinoside, 2'-deoxycoformycin, fludarabine phosphate, phosphate), 5-fluorouracil (5-FU), 5-fluoro-2'-deoxyuridine (5-FUdR), gemcitabine, camptothecin, 6-mercaptopurine, methotrexate, 4-methylthioamphetamine (4-MTA) and thioguanine; and biologics such as interferon-alpha, Bacillus Calmette-Guerin (BCG), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-2 and herceptin.

In addition, the present invention provides a method for preventing, ameliorating and/or treating a disease associated with KRAS mutation in a subject in need thereof, comprising administering to the subject a bifunctional compound described herein or a pharmaceutical composition comprising the same.

In addition, the present invention provides a therapeutically effective amount of a bifunctional compound for use in the manufacture of a medicament for preventing, ameliorating and/or treating a disease associated with a KRAS mutation in a subject in need thereof.

In another aspect, the present invention provides a method for ubiquitination and degradation of a target protein in a cell by contacting the target protein with a bifunctional compound of formula (I) as described herein.

In order to more fully understand the present invention described herein, the following examples are described. The examples described in this application are provided to illustrate the compounds, pharmaceutical compositions and methods provided herein and should not be interpreted as limiting the scope thereof in any way.

Examples

Abbreviation

ACN: acetonitrile; AcOH: acetic acid; Boc: tert-butyloxycarbonyl; DCM: dichloromethane DMA: dimethylacetamide; DMF: dimethylformamide; DMSO: dimethylsulfoxide; DMAC/DMA: dimethylacetamide; DIPEA/DIEA: N,N-diisopropylethylamine; EtOAc/EA: ethyl acetate; EtOH: ethanol; FA: formic acid; HPLC: high pressure liquid chromatography; LAH: lithium aluminum hydroxide; LCMS or LC-MS: liquid chromatography/mass spectrometry; NBS: N-bromosuccinimide; NCS: N-chlorosuccinimide; NMR: nuclear magnetic resonance; NMP: N-methyl-2-pyrrolidone; MeOH: methanol; MPLC: medium pressure liquid chromatography; RT or r.t.: room temperature; TEA: triethylamine; THF: tetrahydrofuran; TFA: trifluoroacetic acid; TLC: thin layer chromatography.

General synthetic method

Synthetic implementation and optimization of bifunctional molecules as described herein can be approached in a stepwise or modular manner. In the presence of RB and ULM, those skilled in the art can combine them using known synthetic methods in the presence or absence of chemical bonding groups. Chemical bonding groups can be synthesized with a range of compositions, lengths, and flexibilities and functionalized so that the RB and ULM groups can be sequentially attached to the ends of the linker. In some cases, protecting group strategies and/or functional group interconversions (FGIs) may be required in order to prepare the desired material. Such chemical processes are well known to synthetic organic chemists and many of these can be found in textbooks or books.

Examples 1 cpd 1 Synthesis

Cpd 1 can be prepared using the synthetic scheme and procedures described in detail below.

(B) Synthesis

Sodium triacetoxyborohydride (NaBH(OAc) ₃ , 32.3 mg, 0.415 mmol, 2 eq) was added to a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(piperidin-1-yl)isoindoline-1,3-dione (Compound A, 100 mg, 0.228 mmol, 1.1 eq) and tributyl 4-(4-methylbenzyl)piperidin-1-carboxylate (63.0 mg, 0.207 mmol, 1.0 eq) in DCM (2.2 ml) and stirred for 2 hours. Upon completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 5% MeOH/DCM) to give the Boc-intermediate (89.2 mg, 62%). To the solution of the Boc-intermediate was added TFA (155 μL, 2.02 mmol, 15 eq). The reaction mixture was stirred at room temperature for 2 hours. Upon completion, the solvent was removed by rotary evaporator and the crude material was purified by silica gel chromatography (0 to 20% MeOH/DCM) to give compound (B), 2-(2,6-dioxopiperidin-3-yl)-5-(4-(4-(piperidin-1-ylmethyl)benzyl)piperidin-1-yl)isoindoline-1,3-dione (71.5 mg, 99%). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.10 (s, 1H), 8.68 (s, 2H), 7.75 (s, 1H), 7.42-7.32 (m, 6H), 5.09 (dd, J = 12.9, 5.4 Hz, 1H), 4.38-4.22 (m, 3H), 3.59 (s, 2H), 3.11 (s, 4H), 2.92-2.85 (m, 1H), 2.62 - 2.52 (m, 6H), 2.06 - 1.98 (m, 1H). LC-MS (m/z): [M] ⁺ calcd. for C ₂₉ H ₃₄ N ₆ O ₄ 530.63, found 531.52.

(D) Synthesis

Compound B (71.5 mg, 0.135 mmol, 1 eq) was added to a solution of tributyl (3-(5-formyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamate (Compound C, 62.4 mg, 0.135 mmol, 1 eq) in DMF (1.3 ml) and stirred for 30 minutes. Sodium triacetoxyborohydride (NaBH(OAc) ₃ , 57.2 mg, 0.27 mmol, 2 eq) was added and stirred for another 16 hours. After completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 7% MeOH/DCM) to give Compound (D), tributyl (3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamate (66.0 mg, 50%). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.09 (s, 1H), 7.82 - 7.73 (m, 3H), 7.67 (d, J = 8.5 Hz, 1H), 7.49 (s, 1H), 7.46 - 7.41 (m, 2H), 7.32 (d, J = 2.2 Hz, 1H), 7.30 - 7.20 (m, 5H), 7.18 (s, 1H), 7.00 (t, J = 5.6 Hz, 1H), 5.07 (dd, J = 12.9, 5.4 Hz, 1H), 4.21 (t, J = 6.9 Hz, 2H), 3.49 (s, 2H), 3.43 (m, 5H), 2.96 - 2.85 (m, 3H), 2.62 - 2.52 (m, 2H), 2.47 (br, 4H) _, 2.36 (br, 4H), 2.04 - 1.98 (m, 1H), 1.91-1.88 (m, 4H), 1.37 (s, 9H). _LC -MS (m/z): [M] ⁺ calculated _{for C53H59F3N8O7 977.10} , found 977.59.

Synthesis of cpd 1

To a solution of compound D (63 mg, 0.0644 mmol) in _CH2Cl2 ( _0.2 mL) was added TFA (100 μL, 1.29 mmol, 20 eq). The reaction mixture was stirred at room temperature for 2 h. Upon completion, the solvent was removed by rotary evaporator and the crude material was purified by silica gel chromatography (0 to 20% MeOH/DCM) to give cpd 1,5-(4-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoleline-1,3-dione (55.9 mg, 98%). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.10 (s, 1H), 7.89-7.69 (m, 8H), 7.46-7.34 (m, 8H), 5.08 (q, J = 6.0 Hz, 1H), 4.45-4.11 (m, 7H), 3.56 (br, 2H), 3.17 (s, 4H), 3.09-3.07 (br, 2H), 2.91-2.89 (m, 2H), 2.82-2.81 (m, 2H), 2.61-2.57 (m, 2H), 2.32 (br, 2H), 2.08 (m, J = 7.2 Hz, 2H), 2.02 (t, J = 5.4 Hz, 1H). LC-MS (m/z): [M] ⁺ calcd. for C ₄₈ H ₅₁ F ₃ N ₈ O ₅ 876.39, found 877.38.

Examples 2 cpd 2 Synthesis

Cpd 2 can be prepared using the synthetic scheme and procedures described in detail below.

(E) Synthesis

1-(tert-Butoxycarbonyl)-4-((tert-Butoxycarbonyl)amino)piperidine-4-carboxylic acid (14 mg, 0.04 mmol) was added along with HBTU (21 mg, 0.056 mmol), DMF (0.4 mL) and DIPEA (0.026 mL, 0.15 mmol), and the solution was stirred at 25 °C for 10 min under Ar atmosphere. 5-(4-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (33 mg, 0.038 mmol) was added, and the resulting mixture was stirred at 25° C. for 2 hours. The reaction mixture was diluted to 10 mL with acetonitrile and purified and concentrated by preparative HPLC to give the title compound (8 mg, 18%). LC-MS (m/z): [M] ⁺ calculated for C ₆₄ H ₇₇ F ₃ N ₁₀ O ₁₀ , 1203.37, found 1203.75. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.11 (s, 1H), 9.98 (s, 1H), 9.42 (s, 1H), 8.03 (s, 1H), 7.89 (s, 1H), 7.77 (m, 3H), 7.69 (s, 1H), 7.46 (m, 6H), 7.37 - 7.27 (m, 2H), 7.07 (s, 1H), 5.10 (dd, J = 12.9, 5.4 Hz, 1H), 4.42 (d, J = 42.2 Hz, 4H), 4.24 (s, 4H), 3.61 (s, 4H), 3.10 (m, 4H), 2.95 - 2.88 (m, 9H).

Synthesis of cpd 2

To a solution of tributyl 4-((tributyloxycarbonyl)amino)-4-((3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)aminocarbonyl) _piperidine -1-carboxylate (5 mg, 0.04 mmol) in _CH2Cl2 (0.1 mL) was added TFA (0.05 mL, 0.62 mmol). The reaction mixture was stirred at room temperature for 1 hour. The mixture was neutralized with _{NaHCO3 (sat)} and diluted with _CH2Cl2 . _{The CH2Cl2 was} dried over _MgSO4 , concentrated, and the crude material was concentrated to give cpd 2,4-amino-N-(3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidine-4-carboxamide (4 mg, 96%). ¹ H NMR (600 MHz, methanol- d ₄ ) δ 8.02 (s, 1H), 7.81 - 7.76 (m, 1H), 7.73 - 7.68 (m, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.38 (m, 4H), 7.26 (dd, J = 8.6, 2.4 Hz, 1H), 5.36 (ddd, J = 5.7, 4.4, 1.1 Hz, 2H), 5.09 (dd, J = 12.8, 5.5 Hz, 1H), 4.64 (s, 6H), 4.34 (t, J = 6.9 Hz, 1H), 3.77 (d, J = 25.0 Hz, 2H), 3.69 - 3.65 (m, 1H), 3.53 (s, 2H), 3.30 - 3.26 (m, 2H), 2.92 - 2.71 (m, 5H), 2.23 - 2.19 (m, 2H), 2.16 - 2.10 (m, 2H), 2.07 _- 2.03 ( _m , 4H), 1.66 - 1.59 (m, 4H). LC-MS (m/z): [M] ⁺ calculated for _{C54H61F3N10O6 , 1003.14} , found 1003.59.

Examples 3 cpd 3 to 72 Synthesis

Cpd 3 can be prepared using the synthetic scheme and procedures described in detail below.

(G) Synthesis

Compound F (70 mg, 0.147 mmol, 1 eq) was added to a solution of tributyl (3-(5-formyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamate (Compound C, 67.9 mg, 0.147 mmol, 1 eq) in DMF (1.4 ml) and stirred for 30 minutes. Sodium triacetoxyborohydride (NaBH(OAc) ₃ , 62.3 mg, 0.294 mmol, 2 eq) was added and stirred for another 16 hours. After completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 5% MeOH/DCM) to give Compound (G) tributyl (3-(5-((4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamate (52.8 mg, 41%). ¹ H NMR (600 MHz, methanol- d ₄ ) δ 7.92 (d, J = 1.5 Hz, 1H), 7.79 - 7.74 (m, 2H), 7.64 (d, J = 8.5 Hz, 1H), 7.60 (s, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.36 - 7.29 (m, 3H), 7.27 (dd, J = 8.5, 1.6 Hz, 1H), 7.18 (dd, J = 8.7, 2.4 Hz, 1H), 5.05 (dd, J = 12.7, 5.5 Hz, 1H), 4.28 (t, J = 6.9 Hz, 2H), 4.03 - 3.95 (m, 3H). 3H), 1.76 - 1.84 (m, 4H), 3.12 - 3.32 (m, 4H), 2.76 - 2.65 (m, 4H), 2.32 (d, J = 6.7 Hz, 2H), 2.13 - 2.06 (m, 1H), 2.03 (p, J = 6.8 Hz, 2H), 1.94 (s, 3H), 1.88-1.85 (m, 3H), 1.44 (s, 9H), 1.30 - 1.22 (m, 2H). LC-MS (m/z): [M] ⁺ _{calcd . for C47H54F3N7O7 885.99} , found 886.45.

Synthesis of cpd 3

To a solution of compound G (40 mg, 0.0451 mmol) in _CH2Cl2 ( _0.15 mL) was added TFA (69.1 μL, 0.903 mmol, 20 eq). The reaction mixture was stirred at room temperature for 2 h. After completion, the solvent was removed by rotary evaporator and the crude material was purified by silica gel chromatography (0 to 20% MeOH/DCM) to give cpd3,5-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (13.0 mg, 37%). ¹ H NMR (600 MHz, MeOD- d ₄ ) δ 7.84 (s, 1H), 7.76-7.74 (m, 2H), 7.63 (d, J = 8.6 Hz, 1H), 7.55 (s, 1H), 7.47 (d, J = 8.6 Hz, 1H), 7.33-7.31 (m, 3H), 7.23 (dd, J = 1.4, 8.5 Hz, 1H), 7.18 (dd, J = 2.4, 8.7 Hz, 1H), 5.05 (dd, J = 5.5, 12.7 Hz, 1H), 4.61 (br, 3H), 4.31 (t, J = 6.9 Hz, 2H), 4.01 (d, J = 13.1 Hz, 2H), 3.66 (br, 2H), 2.97-2.93 (m, 2H), 2.85-2.81 (m, 1H), 2.74-2.68 (m, 4H), 2.54 (br, 6H), 2.23 (d, J = 6.8 Hz, 2H), 2.10-2.04 (m, 3H), 1.88-1.82 (m, 3H), _1.28-1.26 (m, _{3H). LC-MS (m/z): [M]+ calculated for C42H46F3N7O5 785.87} ^, _{found 786.36} .

Examples 4 cpd twenty four Synthesis

Cpd 24 can be prepared using the synthetic scheme and procedures described in detail below.

(H-2) Synthesis

To a solution of compound C (400 mg, 0.865 mmol, 1.1 eq) and ethyl piperidine (129.2 μL, 0.786 mmol, 1.0 eq) in DMF (3.93 mL) was added sodium triacetoxyborohydride (NaBH(OAc) ₃ , 733.3 mg, 3.46 mmol, 4 eq) and stirred at room temperature for 18 hours. After completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 5% MeOH/DCM) to give the ester intermediate (H-2), ethyl 2-(4-((1-(3-((tributyloxycarbonyl)amino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol- ₅ -yl)methyl)piperidin-1-yl)acetate (357.5 mg, 67%). LC-MS (m/z): [M ^]+ _{calcd .} 618.70 _{for C32H41F3N4O5} , found 619.07.

Synthesis of (H-3)

To a solution of compound H-2 (70 mg, 0.113 mmol, 1.0 eq) in THF/EtOH (0.6 mL/0.2 mL) was added a solution of lithium hydroxide (LiOH, 14.2 mg, 0.339 mmol, 3.0 eq) in H ₂ O (0.2 ml) and stirred at room temperature for 4 hours. After completion, the mixture was adjusted to pH 4-5 with 1 N HCl in an ice bath. The solid was collected by filtration and washed with H ₂ O to give (H-3), 2-(4-((1-(3-((tri-butyloxycarbonyl)amino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)acetic acid (37.3 mg, 55.9%). LC-MS (m/z): [M] ⁺ calcd. for C ₃₀ H ₃₇ F ₃ N ₄ O ₅ 590.64, found 590.92.

Synthesis of (H-4)

To a solution of H-3 (37.0 mg, 0.0626 mmol) in DMF (0.3 mL) were added HBTU (28.5 mg, 0.0751 mmol, 1.2 eq) and DIPEA (0.016 mL, 0.113 mmol, 1.8 eq), and the solution was stirred at room temperature under Ar atmosphere for 10 min. 2-(2,6-Dioxopiperidin-3-yl)-5-(piperidin-1-yl)isoindoline-1,3-dione (23.6 mg, 0.0689 mmol, 1.1 eq) was added, and the resulting mixture was stirred at room temperature for 16 h. After completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 6% MeOH/DCM) to give compound (H-4), tributyl (3-(5-((4-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)-2-oxoethyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H _- indol-1-yl)propyl)carbamate (21.3 mg, 37.2%). LC _- MS (m/z): [M] ⁺ calculated _{for C47H53F3N8O8 914.98 ,} found 915.26

Synthesis of cpd 24

To compound H-4 (20 mg, 0.0219 mmol) was added 4N HCl in dioxane (0.24 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted to 1 mL with acetonitrile and purified and concentrated by preparative HPLC to give cpd 24, 5-(4-(2-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)acetyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (4.4 mg, 24.7%). LC-MS (m/z): [M] ⁺ _{calcd. for C42H45F3N8O6 : 814.87 , found} : 815.66. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.04 (d, J = 1.6 Hz, 1H), 7.79 - 7.73 (m, 2H), 7.71 (d, J = 8.5 Hz, 1H), 7.68 - 7.61 (m, 2H), 7.41 - 7.32 (m, 4H), 7.25 (dd, J = 8.6, 2.4 Hz, 1H), 5.08 (dd, J = 12.6, 5.5 Hz, 1H), 4.48 - 4.33 (m, 4H), 3.79 - 3.67 (m, 4H), 3.56 - 3.45 (m, 6H), 3.03 - 2.91 (m, 3H), 2.91-2.81 (m, 2H), 2.79 - 2.65 (m, 2H), 2.24 (p, J = 7.1 Hz, 2H), 2.15-2.07 (m, 1H).

Examples 5 cpd 26 Synthesis

Cpd 26 can be prepared using the synthetic schemes and procedures described in detail below.

(J) Synthesis

To a solution of compound F (90 mg, 0.188 mmol, 1.0 eq.), HBTU (82 mg, 0.216 mmol) and DIPEA (49 mg, 0.376 mmol) in DMF (1.25 mL) was added compound I (121 mg, 0.226 mmol, 1.2 eq.) and stirred for 16 hours. After completion, the solvent was removed in vacuo and the crude material was purified by silica gel chromatography (0 to 5% MeOH/DCM) to give Compound (J) tributyl (3-(5-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidine-1-carbonyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamate ( _{39 mg} , 80%). LC-MS (m/z): [M]+ calculated _{for C47H52F3N7O8 899.97 ,} found 900.59.

Synthesis of cpd 26

To a solution of compound J (55 mg, 0.061 mmol) in ethyl acetate (0.61 mL) was added TFA (1.23 mL, 20 eq). The reaction mixture was stirred at room temperature for 2 h. After completion, the solvent was removed by rotary evaporator and the crude material was purified by silica gel chromatography (0 to 20% MeOH/DCM) to give cpd 26, 5-(4-((1-(1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indole-5-carbonyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (39 mg, 80%). ¹ H NMR (500 MHz, methanol- d ₄ ) δ 7.99 (d, J = 1.1 Hz, 1H), 7.82 - 7.75 (m, 3H), 7.73 (s, 1H), 7.71 - 7.65 (m, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.43 - 7.35 (m, 4H), 5.12 (dd, J = 12.5, 5.5 Hz, 1H), 4.46 (t, J = 6.9 Hz, 2H), 4.19 (d, J = 13.9 Hz, 2H), 3.82 - 3.75 (m, 2H), 3.48 (t, J = 12.9 Hz, 2H), 3.33 - 3.28 (m, 2H), 3.24 (d, J = 6.9 Hz, 2H), 3.03 - 2.96 (m, 2H), 2.95 - 2.84 (m, 1H), 2.82 - 2.68 (m, 2H), 2.39 - 2.25 (m, 3H), 2.19 - 2.10 (m, 1H), 2.04 (s, 1H), 1.92 - 1.88 (m, 1H), 1.51 - 1.36 (m, 2H), 1.27 (t, J = 7.2 _Hz , 1H) _. LC-MS (m/z): [M _] ⁺ calculated for _{C42H44F3N7O6 799.85} , found 800.43.

Example compounds 4-72 in Table 1 were prepared in a manner similar to compound 3 using the corresponding aldehyde starting materials (such as cpd C) and amines (such as cpd F).

surface 1 cpd number Structure and chemical name Mass/NMR 1 5-(4-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₅₃ H ₅₉ F ₃ N ₈ O ₇ 977.10, found 977.59. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.09 (s, 1H), 7.82 - 7.73 (m, 3H), 7.67 (d, J = 8.5 Hz, 1H), 7.49 (s, 1H), 7.46 - 7.41 (m, 2H), 7.32 (d, J = 2.2 Hz, 1H), 7.30 - 7.20 (m, 5H), 7.18 (s, 1H), 7.00 (t, J = 5.6 Hz, 1H), 5.07 (dd, J = 12.9, 5.4 Hz, 1H), 4.21 (t, J = 6.9 Hz, 2H), 3.49 (s, 2H), 3.43 (m, 5H), 2.96 - 2.85 (m, 3H), 2.62 - 2.52 (m, 2H), 2.47 (br, 4H), 2.36 (br, 4H), 2.04 - 1.98 (m, 1H), 1.91-1.88 (m, 4H), 1.37 (s, 9H). 2 4-amino-N-(3-(5-((4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)benzyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidin-4-carboxamide LC-MS (m/z): [M] ⁺ calcd. for C ₅₄ H ₆₁ F ₃ N ₁₀ O ₆ 1003.14, found 1003.59. ¹ H NMR (600 MHz, methanol- d ₄ ) δ 8.02 (s, 1H), 7.81 - 7.76 (m, 1H), 7.73 - 7.68 (m, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.38 (m, 4H), 7.26 (dd, J = 8.6, 2.4 Hz, 1H), 5.36 (ddd, J = 5.7, 4.4, 1.1 Hz, 2H), 5.09 (dd, J = 12.8, 5.5 Hz, 1H), 4.64 (s, 6H), 4.34 (t, J = 6.9 Hz, 1H), 3.77 (d, J = 25.0 Hz, 2H), 3.69 - 3.65 (m, 1H), 3.53 (s, 2H), 3.30 - 3.26 (m, 2H), 2.92 - 2.71 (m, 5H), 2.23 - 2.19 (m, 2H), 2.16 - 2.10 (m, 2H), 2.07 - 2.03 (m, 4H), 1.66 - 1.59 (m, 4H). 3 5-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 786.36. ¹ H NMR (600 MHz, MeOD- d ₄ ) δ 7.84 (s, 1H), 7.76-7.74 (m, 2H), 7.63 (d, J = 8.6 Hz, 1H), 7.55 (s, 1H), 7.47 (d, J = 8.6 Hz, 1H), 7.33-7.31 (m, 3H), 7.23 (dd, J = 1.4, 8.5 Hz, 1H), 7.18 (dd, J = 2.4, 8.7 Hz, 1H), 5.05 (dd, J = 5.5, 12.7 Hz, 1H), 4.61 (br, 3H), 4.31 (t, J = 6.9 Hz, 2H), 4.01 (d, J = 13.1 Hz, 2H), 3.66 (br, 2H), 2.97-2.93 (m, 2H), 2.85-2.81 (m, 1H), 2.74-2.68 (m, 4H), 2.54 (br, 6H), 2.23 (d, J = 6.8 Hz, 2H), 2.10-2.04 (m, 3H), 1.88-1.82 (m, 3H), 1.28-1.26 (m, 3H). 4 5-(4-((4-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₁ H ₄₇ N ₇ O ₅ 718.87, found 718.31. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.01 (d, J = 1.6 Hz, 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.61 (d, J = 8.5 Hz, 1H), 7.50 (m, 3H), 7.36 (m, 2H), 7.22 (dd, J = 8.7, 2.4 Hz, 1H), 6.95 - 6.87 (m, 2H), 5.08 (dd, J = 12.6, 5.5 Hz, 1H), 4.44 - 4.36 (m, 4H), 4.06 (d, J = 13.1 Hz, 2H), 3.08 - 2.94 (m, 6H), 2.87 (m, 2H), 2.80 - 2.70 (m, 2H), 2.63 (d, J = 7.0 Hz, 2H), 2.24 (m, 2H), 2.16 - 2.07 (m, 1H), 1.91 (m, 3H), 1.39 - 1.28 (m, 2H). 5 5-(4-((1-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₁ H ₄₇ N ₇ O ₅ 718.87, found 718.61. ¹ H NMR (600 MHz, methanol- d ₄ ) δ 8.03 (d, J = 1.7 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.53 - 7.47 (m, 4H), 7.36 (ddd, J = 8.5, 4.2, 2.0 Hz, 2H), 6.93 - 6.88 (m, 2H), 5.11 (dd, J = 12.8, 5.4 Hz, 1H), 4.47 (s, 2H), 4.40 (t, J = 6.9 Hz, 2H), 3.59 (d, J = 12.4 Hz, 2H), 3.15 (s, 1H), 3.12 - 3.05 (m, 2H), 3.00 - 2.94 (m, 2H), 2.88 (m, 1H), 2.80 - 2.68 (m, 2H), 2.25 (m, 3H), 2.16 - 2.10 (m, 3H), 1.59 (m, 2H). 6 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 786.27. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 7.86 (d, J = 1.5 Hz, 1H), 7.79 - 7.73 (m, 2H), 7.67 (d, J = 8.5 Hz, 1H), 7.56 (s, 1H), 7.48 (d, J = 8.4 Hz, 1H), 7.34-7.32 (m, 3H), 7.25-7.19 (m, 2H), 5.06 (dd, J = 12.5, 5.4 Hz, 1H), 4.60 (br, 1H), 4.31 (t, J = 7.0 Hz, 2H), 3.68 (s, 2H), 3.44 (t, J = 6.9 Hz, 4H), 2.99 (d, 5H ), 1.83 - 1.76 (m, 2H), 1.65-1.61 (m, 1H), 1.31 - 1.22 (m, 4H). 7 5-(4-((4-((1-(3-aminopropyl)-3-(4-fluorophenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M+H] ⁺ _{calcd. for C41H46FN7O4 , 719.86 ,} found 720.56. ¹ H NMR (500 MHz, MeOD) δ 7.77 (s, 1H), 7.66 - 7.58 (m, 3H), 7.40 - 7.33 (m, 1H), 7.33 - 7.20 (m, 3H), 7.16 - 7.10 (m, 2H), 7.06 (dd, J = 8.7, 2.4 Hz, 1H), 4.27 (td, J = 6.8 Hz, 2H), 4.06 - 3.91 (m, 4H), 3.67 (s, 2H), 3.01 - 2.90 (m, 2H), 2.82 - 2.73 (m, 4H), 2.64 - 2.39 (m, 8H), 2.23 (d, J = 6.8 Hz, 9 - 10 (m, 3H). 8 5-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C36H35F3N6O5 : 688.71 , found} : 689.2. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.11 (d, J = 1.7 Hz, 1H), 7.81 - 7.78 (m, 2H), 7.75 (d, J = 8.5 Hz, 1H), 7.72 - 7.66 (m, 2H), 7.48 - 7.41 (m, 2H), 7.41 - 7.36 (m, 2H), 7.33 (dd, J = 8.6, 2.4 Hz, 1H), 5.10 (dd, J = 12.6, 5.5 Hz, 1H), 4.51 (s, 2H), 4.44 (t, J = 7.0 Hz, 2H), 3.41 (s, 4H), 3.03 - 2.96 (m, δ 5.1 - 5.7 (m, 1H), 2.87 (ddd, J = 17.4, 13.9, 5.2 Hz, 1H), 2.80 - 2.66 (m, 2H), 2.32 - 2.22 (m, 2H), 2.17 - 2.08 (m, 1H). 9 1-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-((1-(2-chloroacetyl)-1,2,3,4-tetrahydroquinolin-6-yl)oxy)ethan-1-one LC-MS (m/z): [M] ⁺ calcd. for C ₃₆ H ₃₉ ClF ₃ N ₅ O ₄ 698.18, found 698.16. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.10 - 8.06 (m, 1H), 7.80 - 7.73 (m, 2H), 7.71 - 7.65 (m, 2H), 7.43 - 7.34 (m, 3H), 6.84 (s, 2H), 4.52 (s, 2H), 4.42 (t, J = 7.0 Hz, 2H), 4.29 (s, 1H), 3.76 (t, J = 6.5 Hz, 2H), 2.97 (t, J = 7.9 Hz, 2H), 2.71 (s, 1H), 2.25 (dt, J = 14.8, 7.1 Hz, 2H), 1.29 (s, 1H). 10 5-(4-((4-((1-(3-aminopropyl)-3-(4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₂ H ₄₉ N ₇ O ₅ 731.90, found 731.30. ¹ H NMR (500 MHz, MeOD) δ 7.91 (d, J = 1.3 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.55 - 7.45 (m, 3H), 7.37 (s, 1H), 7.32 - 7.24 (m, 2H), 7.09 (dd, J = 8.7, 2.4 Hz, 1H), 7.05 - 6.93 (m, 2H), 4.99 - 4.91 (m, 1H), 4.37 - 4.25 (m, 4H), 3.96 (d, J = 13.1 Hz, 2H), 3.84 (s, 3H), 3.29 - 3.16 (m, 3H), 3.01 - 2.87 (m, 6H), 2.80 - 2.70 (m, 3H), 2.53 (d, J = 6.7 Hz, 2H), 2.27 - 2.17 (m, 2H), 2.14 - 2.06 (m, 1H), 1.87 (d, J = 10.8 Hz, 3H), 1.39 - 1.19 (m, 5H). 11 4-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-1H-indol-3-yl)benzonitrile LC-MS (m/z): [M] ⁺ calculated _{for C42H46N8O4 :} 726.88, found _: 727.25. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.16 (d, J = 1.6 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.87 (s, 1H), 7.85 - 7.76 (m, 3H), 7.71 (d, J = 8.5 Hz, 1H), 7.49 (d, J = 2.3 Hz, 1H), 7.44 (dd, J = 8.6, 1.7 Hz, 1H), 7.36 (dd, J = 8.3, 2.3 Hz, 1H), 5.11 (dd, J = 12.5, 5.4 Hz, 1H), 4.52 - 4.42 (m, 3H), 3.78 (s, 4H), 3.61 (d, 3.51 (q, J = 7.0 Hz, 2H), 3.19 (d, J = 7.1 Hz, 1H), 3.11 ( t, J = 12.7 Hz, 2H), 3.03 - 2.97 (m, 2H), 2.94 - 2.83 (m, 1H), 2.81 - 2.67 (m, 2H), 2.27 (m, 2H), 2.17 - 2.10 (m, 2H), 1.63 (m, 2H), 1.20 (t, J = 7.0 Hz, 1H). 12 5-(4-((1-((1-(3-aminopropyl)-3-(6-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₁ H ₄₈ N ₈ O ₅ 732.89, found 733.39. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.38 - 8.33 (m, 1H), 7.91 (dd, J = 8.6, 2.4 Hz, 1H), 7.88 (m, 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.61 (m, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.49 (s, 1H), 7.36 - 7.29 (m, 2H), 7.15 (dd, J = 8.6, 2.3 Hz, 1H), 6.89 (dd, J = 8.6, 0.8 Hz, 1H), 4.98 (dd, J = 12.2, 5.5 Hz, 1H), 4.38 - δ 4.32 (m, 4H), 3.95 (s, 3H), 3.50 (m, 5H), 2.99 - 2.89 (m, 4H), 2.84 - 2.70 (m, 6H), 2.48 (s, 1H), 2.23 (p, J = 7.2 Hz, 2H), 2.14 - 2.07 (m, 1H), 2.05 (d, J = 14.5 Hz, 2H), 1.95 (s, 1H), 1.51-1.44 (m, 2H), 1.24 (m, 1H). 13 3-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-1H-indol-3-yl)benzonitrile LC-MS (m/z): [M] ⁺ _{calcd .} for _{C42H46N8O4 :} 726.88, found: 726.39. ¹ H NMR (600 MHz, MeOD) δ 7.99 (s, 1H), 7.98 - 7.91 (m, 2H), 7.76 (d, J = 8.5 Hz, 1H), 7.64 (s, 1H), 7.63 - 7.60 (m, 2H), 7.60 - 7.57 (m, 2H), 7.40 (d, J = 2.2 Hz, 1H), 7.37 (dd, J = 8.5, 1.5 Hz, 1H), 7.25 (dd, J = 8.5, 2.2 Hz, 1H), 5.13 - 4.97 (m, 1H), 4.43 - 4.35 (m, 4H), 3.72 (s, 3H), 3.52 (d, J = 12.5 Hz, 2H), 3.49 - 3.37 (m, 3H), 3.11 (d, J = 6.6 Hz, 2H), 3.07 - 2.97 (m, 2H), 2.97 - 2.92 (m, 2H), 2.87 - 2.72 (m, 3H), 2.35 - 2.21 (m, 3H), 2.18 - 2.12 (m, 1H), 2.09 (d, J = 13.9 Hz, 2H), 1.70 - 1.59 (m, 2H), 1.26 (s, 2H). 14 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-7-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 786.15. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.00 (d, J = 8.1 Hz, 1H), 7.77 - 7.68 (m, 3H), 7.59 (s, 1H), 7.44-7.43 (m, 1H), 7.40 - 7.34 (m, 3H), 7.34 - 7.26 (m, 2H), 5.09 (dd, J = 12.6, 5.5 Hz, 1H), 4.60 (t, J = 6.8 Hz, 2H), 3.63 (s, 2H), 3.48 (br, 2H), 3.17 (br, 2H), 2.92 - 2.81 (m, 5H), 2.79 - 2.66 (m, 2H), 2.18 - 2.07 (m, 4H), 2.06-2.03 (m, 2H), 1.52 (br, 2H), 1.29 (s, 1H). 15 5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₂ H ₄₈ FN ₇ O ₅ 749.89, found 750.25. ¹ H NMR (500 MHz, DMSO- d ₆ ) δ 11.09 (s, 1H), 7.80 - 7.67 (m, 7H), 7.59 (t, J = 8.8 Hz, 1H), 7.49 (s, 1H), 7.34 (d, J = 8.5 Hz, 1H), 7.00 (dd, J = 12.5, 2.6 Hz, 1H), 6.92 (dd, J = 8.4, 2.6 Hz, 1H), 5.09 (dd, J = 13.0, 5.4 Hz, 1H), 4.42 - 4.33 (m, 4H), 4.22 (s, 1H), 3.83 (s, 3H), 3.61 (s, 2H), 3.18 (m, 3H), 1.77 (s, 2H), 1.11 (t, J = 7.0 Hz , 1H). 16 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-6-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 786.2. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.00 (s, 1H), 7.77 (m, 2H), 7.76 (d, J = 2.1 Hz, 1H), 7.72 (s, 1H), 7.49 (d, J = 2.4 Hz, 1H), 7.40 - 7.36 (m, 2H), 7.31 (dd, J = 8.3, 1.5 Hz, 1H), 5.11 (dd, J = 12.5, 5.5 Hz, 1H), 4.52 - 4.42 (m, 3H), 3.63 (d, J = 12.4 Hz, 2H), 3.51 (m, 2H), 3.20 (d, J = 6.7 Hz, 1H), 3.12 (m, 1H), 3.03 - 2.96 (m, 3H), 2.92 - 2.83 (m, 2H), 2.81 - 2.68 (m, 2H), 2.28 (m, 2H), 2.16 (m, 2H), 1.68 (m, 2H), 1.31 (s, 1H), 1.20 (t, J = 7.0 Hz, 2H). 17 5-(4-((1-((1-(3-aminopropyl)-3-(2-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C41H48N8O5 : 732.89} , found: 733.25. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.09 (dd, J = 5.0, 1.8 Hz, 1H), 7.99 (dd, J = 7.3, 1.9 Hz, 1H), 7.90 (d, J = 1.7 Hz, 1H), 7.77 - 7.71 (m, 2H), 7.65 (d, J = 8.5 Hz, 1H), 7.44 (d, J = 2.3 Hz, 1H), 7.38 (dd, J = 8.6, 1.7 Hz, 1H), 7.31 (dd, J = 8.6, 2.3 Hz, 1H), 7.08 (dd, J = 7.3, 5.0 Hz, 1H), 5.08 (dd, J = 12.6, 1.8 Hz, 1H), 5.11 (d, J = 12.6, 1.8 Hz, 1H), 5.5 Hz, 1H), 4.46 - 4.39 (m, 4H), 3.99 (s, 3H), 3.69 (br, 2H), 3.56 (d, J = 12.4 Hz, 2H), 3.35 (s, 1H), 3.05 (t, J = 12.8 Hz, 2H), 3.01 - 2.94 (m, 3H), 2.87-2.82 (m, 1H), 2.79 - 2.65 (m, 2H), 2.29 - 2.15 (m, 3H), 2.12-2.09 (m, 3H), 1.59-.151 (m, 2H). 18 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethyl)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O4 : 769.87 , found} : 770.15. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.13 (d, J = 1.6 Hz, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.80 (s, 1H), 7.77-7.73 (m, 3H), 7.68 (d, J = 8.5 Hz, 1H), 7.47 (d, J = 2.3 Hz, 1H), 7.41 (dd, J = 8.5, 1.7 Hz, 1H), 7.34 (dd, J = 8.6, 2.4 Hz, 1H), 5.09 (dd, J = 12.6, 5.5 Hz, 1H), 4.48 (s, 2H), 4.43 (t, J = 7.0 Hz, 2H), 3.77 (br, 2H), 3.59 (d, J = 12.5 Hz, 2H), 3.49 (s, 2H), 3.18 (d, J = 6.9 Hz, 2H), 3.13 - 3.04 (m, 2H), 3.02 - 2.95 (m, 2H), 2.92 - 2.81 (m, 1H), 2.79 - 2.65 (m, 2H), 2.29-2.23 (m, 3H), 2.14-2.09 (m, 3H), 1.65-1.57 (m, 2H). 19 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-4-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 787.09. ¹ H NMR (600 MHz, methanol- d ₄ ) δ 7.80 - 7.74 (m, 2H), 7.70 - 7.63 (m, 2H), 7.53 - 7.45 (m, 4H), 7.42 (t, J = 7.8 Hz, 1H), 7.35 (dd, J = 8.4, 2.3 Hz, 1H), 7.31 (d, J = 7.3 Hz, 1H), 5.11 (dd, J = 12.8, 5.5 Hz, 1H), 4.51 - 4.35 (m, 4H), 3.75 (bs, 4H), 3.40 (bs, 4H), 3.09 (d, J = 6.8 Hz, 2H), 3.04 - 2.97 (m, 3H), 1.77 (m, 4H), 2.54 (m, 1H), 2.85 (m, 4H), 2.91 (m, 2H), 2.19 (m, 2H), 2.23 (m, 2H), 2.32 (m, 3H), 2.77 (m, 4H), 2.80 (m, 1H), 2.66 (m, 4H), 2.29 (m, 2H), 2.21 (m, 2H), 2.17 (m, 2H), 1.94 (d, J = 14.3 Hz, 2H), 1.43 (m, 2H). 20 4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 785.87. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.06 (d, J = 1.7 Hz, 1H), 7.78 - 7.74 (m, 2H), 7.73 - 7.64 (m, 3H), 7.45 (d, J = 7.2 Hz, 1H), 7.40 - 7.32 (m, 4H), 5.10 (dd, J = 12.7, 5.5 Hz, 1H), 4.44-4.40 (m, 4H), 3.55-3.53 (m, 4H), 3.05 (t, J = 12.7 Hz, 2H), 3.00 - 2.93 (m, 3H), 2.90 - 2.81 (m, 2H), 2.78 - 2.66 (m, 2H), 2.29 - 2.22 (m, 2H), 2.14-2.08 (m, 3H), 1.56-1.48 (m, 1H). twenty one (3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamic acid tributyl ester LC-MS (m/z): [M] ⁺ _{calcd. for C47H54F3N7O7 885.99 , found 886.31} . ¹ H NMR (600 MHz, methanol- d ₄ ) δ 8.03 (d, J = 1.6 Hz, 1H), 7.79 - 7.74 (m, 2H), 7.66 (t, J = 4.3 Hz, 2H), 7.59 (d, J = 8.5 Hz, 1H), 7.36 - 7.31 (m, 4H), 7.20 (dd, J = 8.6, 2.4 Hz, 1H), 5.06 (dd, J = 12.8, 5.4 Hz, 1H), 4.61 (br, 2H), 4.40 (s, 2H), 4.30 (t, J = 6.9 Hz, 2H), 3.48 (s, 2H), 3.43 (t, J = 5.0, 4H), 3.34 (s, 2H), 3.06 (t, J = 6.7 Hz, 2H), 3.00 (br, 2H), 2.89 - 2.80 (m, 1H), 2.76 - 2.64 (m, 2H), 2.57 (t, J = 5.1 Hz, 4H), 2.29-2.28 (m, 2H), 2.12 - 1.99 (m, 5H), 1.91 (s, 1H), 1.43 (s, 9H). twenty two 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)prop-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₀ H ₃₈ F ₃ N ₅ O ₆ 741.77, found 743.67. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 11.15 (d, J = 2.3 Hz, 1H), 9.39 (s, 1H), 8.07 (dd, J = 24.9, 1.6 Hz, 1H), 7.93 (dd, J = 7.7, 5.1 Hz, 1H), 7.90 (d, J = 2.9 Hz, 1H), 7.82 - 7.68 (m, 8H), 7.50 - 7.45 (m, 2H), 7.36 (ddd, J = 25.9, 8.5, 1.6 Hz, 1H), 6.82 - 6.75 (m, 1H), 6.06 (m, H), 5.17 (m, 1H), 4.83 (dd, J = 3.1, 1.6 Hz, 1H), 7 - 11 (m, 3H), 3.57 (m, 1H), 3.20 (d, J = 12.3 Hz, 1H), 3.19 (m, 1H), 3.35 (m, 3H), 3.23 (m, 1H), 3.13 (m, 2H), 3.21 (d, J = 12.3 Hz, 1H), 3.19 (m, 1H), 3.30 ( m, 3H), 3.23 (m, 1H), 3.26 (m, 2H), 3.13 (m, 1H), 3.20 (m, 2H), 3.31 (m, 3H), 3.30 ( m , 1H), 3.33 (m, 3H), 3.36 (m, 1H), 1.10 (t, J = 7.0 Hz, 1H). twenty three 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)propyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd .} for _{C40H42F3N5O6 : 745.80} , found: 746.48. ¹ H NMR (600 MHz, methanol- d ₄ ) δ 8.09 (s, 1H), 7.83 - 7.77 (m, 4H), 7.70 (m, 2H), 7.68 - 7.62 (m, 3H), 7.45 - 7.35 (m, 4H), 5.18 - 5.15 (m, 1H), 4.46 (d, J = 2.8 Hz, 2H), 4.44 (t, J = 7.4 Hz, 1H), 4.36 (t, J = 7.1 Hz, 2H), 3.69 (s, 1H), 3.55 - 3.51 (m, 2H), 3.49 (t, J = 5.8 Hz, 2H), 3.24 - 3.18 (m, 2H), 3.07 (t, J = 12.6 Hz, 1H), 3.00 - 2.96 (m, 2H), 2.92 - 2.86 (m, 2H), 2.83 - 2.74 (m, 4H), 2.25 (m, 4H), 2.16 (m, 1H), 2.09 (d, J = 15.5 Hz, 2H), 2.00 - 1.91 (m, 3H), 1.86 (t, J = 14.1 Hz, 2H), 1.66 (m, 1H). twenty four 5-(4-(2-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)acetyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [ _M ] ⁺ calculated _{for C42H45F3N8O6 :} 814.87, found _: 815.66. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 8.04 (d, J = 1.6 Hz, 1H), 7.79 - 7.73 (m, 2H), 7.71 (d, J = 8.5 Hz, 1H), 7.68 - 7.61 (m, 2H), 7.41 - 7.32 (m, 4H), 7.25 (dd, J = 8.6, 2.4 Hz, 1H), 5.08 (dd, J = 12.6, 5.5 Hz, 1H), 4.48 - 4.33 (m, 4H), 3.79 - 3.67 (m, 4H), 3.56 - 3.45 (m, 6H), 3.03 - 2.91 (m, 3H), 2.91-2.81 (m, 2H), 2.79 - 2.65 (m, 2H), 2.24 (p, J = 7.1 Hz, 2H), 2.15-2.07 (m, 1H). 25    5-(4-((1-((1-(3-aminopropyl)-3-(3-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd .} for _{C42H46F3N7O5 : 785.35} , found: 786.34. ¹ H NMR (600 MHz, methanol-d4) δ 8.13 - 8.11 (m, 1H), 7.79 - 7.77 (m, 3H), 7.72 (dd, J = 8.6, 0.6 Hz, 1H), 7.60 - 7.57 (m, 2H), 7.50 (m, 2H), 7.37 (dd, J = 8.6, 2.4 Hz, 1H), 7.23 (ddt, J = 8.2, 2.3, 1.1 Hz, 1H), 5.13 - 5.10 (m, 1H), 4.51 (s, 2H), 4.46 (t, J = 7.0 Hz, 2H), 4.26 - 4.13 (m, 2H), 3.77 (s, 2H), 3.59 (d, J = 12.4 Hz, 2H), 3.53 - 3.43 (m, 2H), 3.39 - 3.35 (m, 1H), 3.29 (s, 1H), 3.24 - 3.20 (m, 2H), 3.15 (m, 2H), 3.01 -2.97 (m, 2H), 2.88 (m, 1H), 2.78 (m, 2H), 2.34 (m, 1H), 2.30 - 2.25 (m, 2H), 2.21 - 2.16 (m, 2H), 2.16 -2.11 (m, 1H), 1.72 - 1.63 (m, 2H). 26    5-(4-((1-(1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indole-5-carbonyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [ _M ] ⁺ calculated for _{C42H44F3N7O6 : 799.85} , found _: 800.43. ¹ H NMR (500 MHz, methanol- d ₄ ) δ 7.99 (d, J = 1.1 Hz, 1H), 7.82 - 7.75 (m, 3H), 7.73 (s, 1H), 7.71 - 7.65 (m, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.43 - 7.35 (m, 4H), 5.12 (dd, J = 12.5, 5.5 Hz, 1H), 4.46 (t, J = 6.9 Hz, 2H), 4.19 (d, J = 13.9 Hz, 2H), 3.82 - 3.75 (m, 2H), 3.48 (t, J = 12.9 Hz, 2H), 3.33 - 3.28 (m, 2H), 3.24 (d, J = 6.9 Hz, 2H), 3.03 - 2.96 (m, 2H), 2.95 - 2.84 (m, 1H), 2.82 - 2.68 (m, 2H), 2.39 - 2.25 (m, 3H), 2.19 - 2.10 (m, 1H), 2.04 (s, 1H), 1.92 - 1.88 (m, 1H), 1.51 - 1.36 (m, 2H), 1.27 (t, J = 7.2 Hz, 1H). 27    5-(4-((1-((1-(3-aminopropyl)-3-(3-hydroxynaphthalen-1-yl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₅ H ₄₉ N ₇ O ₅ 767.93, found 768.25. ¹ H NMR (600 MHz, MeOD) δ 7.84 (d, J = 8.5 Hz, 1H), 7.76 (dd, J = 17.6, 8.7 Hz, 3H), 7.64 (s, 1H), 7.60 (s, 1H), 7.51 - 7.47 (m, 2H), 7.42 (t, J = 7.5 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.25 - 7.20 (m, 2H), 7.19 (d, J = 2.2 Hz, 1H), 5.12 (dd, J = 12.8, 5.4 Hz, 1H), 4.53 (t, J = 6.9 Hz, 2H), 4.41 (s, 2H), 4.21 - 4.13 (m, 2H), 3.80 - 3.71 (m, 2H), 3.59 - 3.42 (m, 4H), 3.31 - 3.23 (m, 2H), 3.20 (d, J = 6.8 Hz, 2H), 3.11 - 3.00 (m, 4H), 2.93 - 2.84 (m, 1H), 2.81 - 2.66 (m, 2H), 2.40 - 2.22 (m, 3H), 2.21 - 2.08 (m, 3H), 1.70 - 1.54 (m, 2H) 28    5-(4-((1-((1-(Azocyclobutane-3-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M]+ _{calcd. for C43H46F3N7O5 : 797.88 , found} : 798.65. ¹ H NMR (600 MHz, methanol-d ₄ ) δ 8.16 (d, J = 1.6 Hz, 1H), 7.87 - 7.80 (m, 2H), 7.79 - 7.72 (m, 3H), 7.52 - 7.46 (m, 2H), 7.40 (d, J = 8.2 Hz, 2H), 7.38 - 7.33 (m, 1H), 5.11 (dd, J = 12.8, 5.4 Hz, 1H), 4.55 - 4.44 (m, 4H), 3.78 (dd, J = 12.1, 3.8 Hz, 1.39 - 1.24 (m, 8H), 0.92 (t, J = 7.0 Hz, 1H). 3.54 (m, 3H), 3.29 (dd, J = 13.3, 8.4 Hz, 2H), 3.15 (t, J = 12.7 Hz, 2H), 3.10 (dd, J = 13.3, 5.0 Hz, 2H), 2.88 (ddd, J = 17.5, 14.4, 5.6 Hz, 2H), 2.80 - 2.68 (m, 2H), 2.20 - 2.10 (m, 3H), 2.05 (q, J = 6.5 Hz, 1H), 1.73 - 1.59 (m, 2H), 1H). 29    5-(4-((1-((1-(3-aminopropyl)-3-(5-chloro-6-methyl- 1H -indazol-4-yl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₃ H ₄₈ ClN ₉ O ₄ 790.37, found 790.40. ¹ H NMR (600 MHz, MeOD) δ 7.86 - 7.75 (m, 3H), 7.73 (s, 1H), 7.59 - 7.47 (m, 4H), 7.37 (dd, J = 8.5, 2.2 Hz, 1H), 5.12 (dd, J = 12.8, 5.5 Hz, 1H), 4.59 - 4.49 (m, 2H), 4.43 (s, 2H), 4.24 - 4.12 (m, 2H), 3.84 - 3.71 (m, 2H) , 3.60 - 3.43 (m, 4H), 3.21 - 3.24 (m, 2H), 3.26 - : 2.98 (m, 4H), 2.94 - 2.85 (m, 1H), 2.81 - 2.69 (m, 2H), 2.61 (s, 3H), 2.39 - 2.25 (m, 3H), 2.21 - 2.10 (m, 3H), 1.71 - 1.56 (m, 2H). 30    5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd. for C42H46F3N7O5 : 785.87 , found} : 786.20. ¹ H NMR (600 MHz, MeOD) δ 8.13 (d, J = 1.1 Hz, 1H), 7.83 - 7.80 (m, 2H), 7.75 (d, J = 8.5 Hz, 1H), 7.72 - 7.66 (m, 2H), 7.46 (dd, J = 8.6, 1.7 Hz, 2H), 7.39 - 7.32 (m, 3H), 5.09 (dd, J = 12.7, 5.5 Hz, 1H), 4.48 (s, 2H), 4.43 (t, J = 7.0 Hz, 2H), 4.15 (d, J = 12.6 Hz, 2H), 3.75 (br, 2H), 3.56 (d, J = 12.7 Hz, 3.51 (m, 2H), 3.57 (s, 1H), 3.25 (s, 1H), 3.19 (d, J = 6.3 Hz, 2H), 3.25 (t, J = 12.0 Hz, 2H), 3.30 (m, 2H), 3.74 (m, 1H), 3.85 (m, 2H), 3.14 (t, J = 12.0 Hz, 2H), 3.24 (m, 1H), 3.55 (m, 2H), 3.20 (d, J = 6.3 Hz, 2H), 3.19 (t, J = 12.0 Hz, 2H), 3.23 (m, 1H), 3.84 (m, 1H), 3.91 (m, 2H). 31    (2S)-N-(2-(3-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carboxamide LC-MS (m/z): [M] ⁺ calculated _{for C52H64F4N8O6S : 1005.19 ,} found _: 1005.6. ¹ H NMR (600 MHz, DMSO) δ 8.99 (s, 1H), 8.55 - 8.48 (m, 1H), 7.87 - 7.69 (m, 7H), 7.58 (d, J = 8.3 Hz, 1H), 7.45 (d, J = 8.2 Hz, 2H), 7.40 (s, 1H), 7.27 (d, J = 8.9 Hz, 1H), 6.99 (s, 2H), 5.19 (d, J = 3.5 Hz, 1H), 4.58 (d, J = 9.2 Hz, 1H), 4.50 (t, J = 8.2 Hz, 1H), 4.43 (s, 1H), 4.34 (s, 3H), 4.26 (s, 7 - 1.47 (m, 2H), 3.54 - 3.48 (m, 1H), 3.10 - 2.99 (m, 3H), 2.98 - 2.91 (m, 1H), 2.81 (s, 2H), 2.45 (s, 3H), 2.34 (s, 1H), 2.30 - 2.23 (m, 1H), 2.13 (s, 1H), 2.11 - 2.03 (m, 3H), 1.93 - 1.84 (m, 2H), 1.39 - 1.30 (m, 2H), 1.25 - 1.19 (m, 3H), 0.96 (s, 9H). 32    5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd .} for _{C42H45F4N7O5 : 803.86} , found: 804.30. ¹ H NMR (600 MHz, MeOD) δ 8.16 (s, 1H), 7.84 (d, J = 8.6 Hz, 2H), 7.74 (s, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.67 (d, J = 10.7 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 7.49 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 8.3 Hz, 2H), 5.14 (dd, J = 12.9, 5.4 Hz, 1H), 4.51 (s, 2H), 4.46 (t, J = 6.9 Hz, 2H), 3.82 (dd, J = 32.8, 12.4 Hz, 4H), 3.60 (d, J = 12.8 Hz, 2H), 3.49 (t, J = 12.4 Hz, 2H), 3.41 - 3.35 (m, 2H), 3.25 (d, J = 6.7 Hz, 2H), 3.16 (t, J = 12.1 Hz, 2H), 3.04 - 2.97 (m, 2H), 2.93 - 2.86 (m, 1H), 2.82 - 2.69 (m, 2H), 2.40 - 2.32 (m, 1H), 2.31 - 2.25 (m, 2H), 2.22 - 2.10 (m, 3H), 1.76 - 1.66 (m, 2H). 33    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(methylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M]+ _{calcd .} for _{C43H46F3N7O5 : 799.90} , found: 800.60. ¹ H NMR (600 MHz, MeOD) δ 8.14 (s, 1H), 7.82 (d, J = 8.6 Hz, 2H), 7.77 - 7.71 (m, 2H), 7.69 (d, J = 8.5 Hz, 1H), 7.50 - 7.44 (m, 2H), 7.39 - 7.32 (m, 3H), 5.09 (dd, J = 12.8, 5.4 Hz, 1H), 4.48 (s, 2H), 4.44 (t, J = 6.9 Hz, 2H), 4.15 (d, J = 14.2 Hz, 2H), 3.76 (d, J = 11.4 Hz, 2H), 3.56 (d, J = 12.9 Hz, 2H), 3.51 - 3.43 (m, 2H), 3.27 (t, J = 11.2 Hz, 2H), 3.21 (d, J = 6.7 Hz, 2H), 3.14 (t, J = 12.1 Hz, 2H), 3.07 - 3.01 (m, 2H), 2.90 - 2.82 (m, 1H), 2.77 - 2.70 (m, 2H), 2.37 - 2.25 (m, 3H), 2.17 (d, J = 13.9 Hz, 2H), 2.13 - 2.08 (m, 1H), 1.74 - 1.64 (m, 2H). 34    5-(7-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₃₉ H ₃₉ F ₃ N ₆ O ₅ 728.77, found 729.1. ¹ H NMR (600 MHz, MeOD) δ 8.10 (d, J = 5.2 Hz, 1H), 7.80 - 7.79 (m, 2H), 7.71 - 7.69 (m, 2H), 7.55 (dd, J = 8.4, 2.8 Hz, 1H), 7.37 (t, J = 7.1 Hz, 4H), 7.10 (d, J = 2.1 Hz, 1H), 6.98 (dd, J = 8.4, 2.2 Hz, 1H), 6.84 (d, J = 2.0 Hz, 1H), 6.68 (dd, J = 8.3, 2.1 Hz, 1H), 5.07 - 5.02 (m, 2H), 4.47 (d, J = 12.5 Hz, 3H), 3.93 (d, J = 15.8 Hz, 4H), 3.49 (d, J = 13.1 Hz, 1H), 3.17 - 3.11 (m, 2H), 3.01 - 2.96 (m, 4H), 2.86 - 2.83 (m, 1H), 2.74 - 2.70 (m, 2H), 2.26 - 2.24 (m, 3H), 2.09 - 2.07 (m, 2H). 35    5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-((3-fluoropyridin-2-yl)methoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M _] ⁺ calculated for _{C47H50F2N8O5 : 844.97} , found _: 845.6. ¹ H NMR (600 MHz, MeOD) δ 8.45 (d, J = 4.7 Hz, 1H), 7.86 (s, 1H), 7.71-7.75 (m, 2H), 7.64 (d, J = 8.5 Hz, 1H), 7.57-7.61 (m, 2H), 7.51-7.54 (m, 1H), 7.45 (s, 1H), 7.37 (dd, J = 1.2, 8.5 Hz, 1H), 7.32-7.33 (m, 1H), 6.95-6.99 (m, 2H), 5.30 (d, J = 1.8 Hz, 2H), 5.09 (dd, J = 5.1, 12.7 Hz, 1H), 4.39-4.43 (m, 4H), 3.98 (s, 2H), 3.80 (br, 2H), 3.53-3.56 (m, 4H), 3.49 (br, 2H), 3.17 (d, J = 6.8 Hz, 2H), 3.04-3.08 (m, 2H), 2.95-2.97 (m, 2H), 2.82-2.88 (m, 1H), 2.67-2.75 (m, 2H), 2.21-2.27 (m, 3H), 2.09-2.11 (m, 3H), 1.55-1.62 (m, 2H). 36    5-(4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)azocyclobutan-3-yl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₃₉ H ₄₀ F ₃ N ₇ O ₅ 743.79, found 744.50. ¹ H NMR (600 MHz, MeOD) δ 8.09 (s, 1H), 7.83 - 7.79 (m, 2H), 7.75 - 7.70 (m, 2H), 7.69 (d, J = 8.5 Hz, 1H), 7.44 - 7.37 (m, 4H), 7.28 (dd, J = 8.6, 2.2 Hz, 1H), 5.11 (dd, J = 12.8, 5.5 Hz, 1H), 4.57 (s, 2H), 4.44 (t, J = 7.0 Hz, 2H), 4.30 - 4.23 (m, 2H), 4.20 - 4.13 (m, 2H), 3.56 - 3.50 (m, 4H), 3.46 (p, J = 6.7 Hz, 1H), 3.03 - 2.97 (m, 2H), 2.93 - 2.85 (m, 1H), 2.81 - 2.69 (m, 2H), 2.67 - 2.59 (m, 4H), 2.31 - 2.23 (m, 2H), 2.16 - 2.10 (m, 1H). 37    5-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M _] ⁺ calculated for _{C44H50F3N7O5 : 813.92} , found _: 814.4. ¹ H NMR (600 MHz, MeOD) δ 7.91 (d, J = 0.9 Hz, 1H), 7.79 - 7.73 (m, 2H), 7.66 (d, J = 8.5 Hz, 1H), 7.59 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.36 - 7.31 (m, 3H), 7.27 (dd, J = 8.5, 1.5 Hz, 1H), 7.20 (dd, J = 8.6, 2.3 Hz, 1H), 5.06 (dd, J = 12.7, 5.5 Hz, 1H), 4.30 (t, J = 6.9 Hz, 2H), 3.87 (s, 2H), 3.46 - 3.39 3.15 - 3.07 (m, 2H), 2.88 - 2.81 (m, 1H), 2.76 - 2.67 (m, 2H), 2.59 - 2.52 (m, 4H), 2.39 - 2.30 (m, 4H), 2.26 (s, 6H), 2.11 - 2.05 (m, 3H), 1.86 (d, J = 12.4 Hz, 2H), 1.72 - 1.66 (m, 1H), 1.35 - 1.27 (m, 4H). 38    5-(4-((1-((1-(5-aminopentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C _{4 4} H ₅₀ F ₃ N ₇ O ₅ 813.38; found: 814. ¹ H NMR (600 MHz, DMSO ): δ 11.11 (s, 1H), 8.05 (s, 1H), 7.89 (s, 1H), 7.81-7.76 (m, 3H), 7.75-7.66 (m, 3H), 7.49 (s, 1H), 7.46 (d, J = 7.8 Hz, 2H), 7.39-7.32 (m, 2H), 5.08 (dd, J = 13.2 Hz, 5.4 Hz, 1H), 4.18 (s, 1H), 4.25 (t, J = 6.6 Hz, 2H), 3.06 (m, 5H), 2.95-2.88(m, 3H), 2.78 (m, 2H), 2.02 (m, 7 (m, 2H), 1.97 (m, 2H), 1.84 (m, 2H), 1.58 (m, 2H), 1.33 (m, 4H), 1.23 (m, 4H). 39    5-(7-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₅ H ₅₀ F ₃ N ₇ O ₅ 825.93, found 825.50. ¹ H NMR (600 MHz, MeOD) δ 8.09 (d, J = 1.2 Hz, 1H), 7.82 - 7.76 (m, 2H), 7.72 (s, 1H), 7.68 (dd, J = 8.4, 4.7 Hz, 2H), 7.42 (dd, J = 8.5, 1.6 Hz, 1H), 7.39 (d, J = 8.0 Hz, 2H), 6.88 (d, J = 2.1 Hz, 1H), 6.72 (dd, J = 8.4, 2.1 Hz, 1H), 5.09 (dd, J = 12.7, 5.5 Hz, 1H), 4.49 (s, 2H), 4.45 (t, J = 7.0 Hz, 2H), 3.92 (d, J = 47.5 Hz, 4H), 3.70 - 3.54 (m, 4H), 3.17 - 3.04 (m, 5H), 3.03 - 2.97 (m, 2H), 2.92 - 2.84 (m, 1H), 2.80 - 2.68 (m, 2H), 2.36 - 2.21 (m, 5H), 2.21 - 2.07 (m, 5H), 1.67 - 1.56 (m, 2H). 40    N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)acetamide LC-MS (m/z): [M] ⁺ _{calcd .} for _{C44H48F3N7O6 : 827.91} , found: 828.7. ¹ H NMR (600 MHz, MeOD) δ 8.02 (d, J = 1.0 Hz, 1H), 7.81 - 7.76 (m, 2H), 7.69 - 7.65 (m, 2H), 7.59 (d, J = 8.5 Hz, 1H), 7.37 - 7.31 (m, 4H), 7.21 (dd, J = 8.6, 2.3 Hz, 1H), 5.07 (dd, J = 12.7, 5.5 Hz, 1H), 4.34 - 4.26 (m, 4H), 3.47 - 3.36 (m, 6H), 3.22 - 3.17 (m, 3H), 2.93 - 2.80 (m, 3H), 2.77 - 2.67 (m, 2H), 2.61 - 2.53 (m, 4H), 2.29 (d, J = 7.1 Hz, 2H), 2.13 - 2.06 (m, 3H), 2.02 (d, J = 13.5 Hz, 2H), 1.93 (s, 3H), 1.88 (br, 1H), 1.46 - 1.36 (m, 2H). 41    4-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)butyronitrile LC-MS (m/z): [M] ⁺ _{calcd. for C43H44F3N7O5 : 795.86 , found} : 796.4. ¹ H NMR (600 MHz, MeOD) δ 8.05 (d, J = 1.3 Hz, 1H), 7.80 - 7.75 (m, 2H), 7.69 - 7.63 (m, 3H), 7.40 - 7.31 (m, 4H), 7.22 (dd, J = 8.6, 2.4 Hz, 1H), 5.07 (dd, J = 12.7, 5.5 Hz, 1H), 4.44 - 4.38 (m, 4H), 3.52 - 3.40 (m, 6H), 3.01 (t, J = 10.4 Hz, 2H), 2.88 - 2.81 (m, 1H), 2.76 - 2.67 (m, 2H), 2.63 - 2.54 (m, 4H), 2.46 (t, J = 7.1 Hz, 2H), 2.30 (d, J = 6.9 Hz, 2H), 2.24 (p, J = 7.0 Hz, 2H), 2.12 - 2.02 (m, 3H), 1.94 - 1.87 (m, 1H), 1.43 (br, 2H). 42    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-methyl-1H-imidazol-5-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M _] ⁺ calculated _{for C44H45F3N8O5 : 822.35} , found: 823.8. ¹ H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 8.01 (br, 1H), 7.87 (s, 1H), 7.78 (d, J = 9.0 Hz, 2H), 7.75 (br. 1H), 7.67 (d, J = 8.4 Hz, 1H),7.59 (s, 1H), 7.46 (d, J = 8.4 Hz, 2H), 7.33 (d, 2H), 7.25 (dd, J = 7.2 Hz, 1.8 Hz, 1H), 7.05 (s, 1H), 5.52 (s, 2H), 5.06 (dd, J = 13.2 Hz, 5.4 Hz, 1H), 4.36 (br, 1H), 3.55 (s, 3H), 2.87 (m, 2H), 2.47 (br, 4H), 2.16 (br. 2H), 2.01 (m, 2H), 1.91 (s, 4H), 1.76 (br, 2H), 1.23 (br, 4H). 43    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-methoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M _] ⁺ calculated for _{C43H47F3N6O6 : 800.88} , found _: 801.9. ¹ H NMR (600 MHz, MeOD) δ 7.93 (s, 1H), 7.75-7.78 (m, 2H), 7.66 (d, J = 8.3 Hz, 1H), 7.56 (s, 1H), 7.51 (d, J = 8.3 Hz, 1H), 7.33-7.34 (m, 3H), 7.27 (dd, J = 1.3, 8.3 Hz, 1H), 7.20 (dd, J = 2.3, 8.6 Hz, 1H), 5.06 (dd, J = 5.3, 12.6 Hz, 1H), 4.34 (t, J = 6.9 Hz, 2H), 3.97 (br, 2H), 3.42-3.44 (m, 4H), 3.32 (s, 3H), 3.17-3.21 (m, 2H), 2.82-2.88 (m, 1H), 2.66-2.75 (m, 2H), 2.55-2.57 (m, 4H), 2.47 (br, 2H), 2.27 (d, J = 7.3 Hz, 2H), 2.07-2.12 (m, 3H), 1.89-1.91 (m, 2H), 1.74-1.77 (m, 1H), 1.28-1.36 (m, 4H) 44    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(2,2,2-trifluoroethoxy)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C44H46F6N6O6 :} 868.34; found: 869.5. ¹ H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 8.01 (br, 1H), 7.84 (s, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.68 (d, J = 8.4 Hz, 1H), 7.61 (br., 1H), 7.46 (d, J = 8.4 Hz, 2H), 7.33 (m, 2H), 7.25 (d, J = 9.0 Hz, 1H), 5.05 (dd, J = 12.6 Hz, 5.4 Hz, 1H), 4.31 (t, J = 6.6 Hz, 2H), 4.05 (q, J = 9.6 Hz, 2H), 3.58 (t, J = 6.6 Hz, 3H), 2.88 (m, 2H), 2.47 (m, 4H), 2.17 (br,2H), 2.10 (m, 4H), 2.02 (m, 2H), 1.91 (s, 4H), 1.76 (br, 2H), 1.23 (br, 4H). 45    N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-1H-pyrrole-2-carboxamide LC-MS (m/z): [M] ⁺ _{calcd . for C47H49F3N8O6 : 878.95 ,} found: 879.6. ¹ H NMR (600 MHz, MeOD) δ 8.04 (d, J = 1.2 Hz, 1H), 7.78 - 7.75 (m, 2H), 7.72 - 7.71 (m, 1H), 7.70 - 7.66 (m, 1H), 7.61 (d, J = 8.5 Hz, 1H), 7.35 - 7.34 (m, 2H), 6.90 (dd, J = 2.5, 1.4 Hz, 1H), 6.74 (dd, J = 3.7, 1.4 Hz, 1H), 6.15 (dd, J = 3.7, 2.6 Hz, 1H), 5.07 (dd, J = 12.7, 5.5 Hz, 1H), 4.42 (s, 2H), δ 4.37 (t, J = 6.9 Hz, 2H), 3.72 (dt, J = 13.3, 6.6 Hz, 4H), 3.51 (br, 1H), 3.47 - 3.43 (m, 4H), 3.38 (t, J = 6.8 Hz, 2H), 3.23 (q, J = 7.4 Hz, 4H), 2.88 - 2.82 (m, 1H), 2.77 - 2.67 (m, 2H), 2.62 (s, 4H), 2.17 (p, J = 6.8 Hz, 2H), 2.13 - 2.03 (m, 3H). 46    3-((4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M]+ calculated _{for C35H37F4N5O3 : 651.71} , found _: 652.3. ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 10.81 (s, 1H), 9.55 (s, 1H), 8.08 (s, 1H), 7.91 (s, 1H), 7.81 - 7.79 (m, 3H), 7.73 (d, J = 8.5 Hz, 1H), 7.50 - 7.46 (m, 2H), 7.39 - 7.36 (m, 1H), 6.92 (t, J = 8.8 Hz, 1H), 6.49 - 6.43 (m, 2H), 6.11 (d, J = 7.8 Hz, 1H), 4.44 (s, 2H), 4.36 (t, J = 6.9 Hz, 2H), 4.31 (m, 3.43 (d, J = 11.7 Hz, 2H), 3.11 (d, J = 11.5 Hz, 2H), 2.89 (s, 1H), 2.85 - 2.80 (m, 2H), 2.73 (m, 1H), 2.07 (m, 4H), 1.92 - 1.82 (m, 5H), 1.24 (d, J = 3.3 Hz, 2H). 47    N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-3-(trifluoromethyl)benzamide LC-MS (m/z): [M] ⁺ calcd. for C ₅₀ H ₄₉ F ₆ N ₇ O ₆ 957.98, found 958.4. ¹ H NMR (500 MHz, MeOD) δ 8.07 (s, 2H), 8.00 (d, J = 7.8 Hz, 1H), 7.83 (d, J = 7.8 Hz, 1H), 7.78 - 7.74 (m, 3H), 7.73 (s, 1H), 7.67 - 7.61 (m, 2H), 7.47 (d, J = 2.3 Hz, 1H), 7.38 (dd, J = 8.5, 1.6 Hz, 1H), 7.36 - 7.30 (m, 3H), 5.09 (dd, J = 12.6, 5.5 Hz, 1H), 4.46 (s, 2H), 4.41 (t, J = 6.7 Hz, 2H), 4.14 3.74 (s, 2H), 3.57 (d, J = 12.8 Hz, 2H), 3.49 - 3.39 (m, 4H), 3.35 (s, 2H), 3.20 (d, J = 6.8 Hz, 2H), 3.11 (t, J = 12.0 Hz, 2H), 2.89 - 2.82 (m, 1H), 2.78 - 2.67 (m, 2H), 2.33 - 2.22 (m, 3H), 2.19 - 2.07 (m, 3H), 1.70 - 1.58 (m, 2H). 48    N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-4-fluorobenzamide LC-MS (m/z): [M _] ⁺ calculated for _{C49H49F4N7O6 : 907.97} , found _: 908.4. ¹ H NMR (600 MHz, MeOD) δ 8.08 (d, J = 1.1 Hz, 1H), 7.83 - 7.72 (m, 6H), 7.63 (d, J = 8.5 Hz, 1H), 7.46 (d, J = 2.2 Hz, 1H), 7.39 - 7.32 (m, 4H), 7.19 - 7.12 (m, 2H), 5.09 (dd, J = 12.8, 5.5 Hz, 1H), 4.46 (s, 2H), 4.40 (t, J = 6.8 Hz, 2H), 4.20 - 4.10 (m, 2H), 3.74 (br, 2H), 3.56 (d, J = 12.8 Hz, 2H), 3.43 (t, J = 6.8 Hz, 4H), 3.29 - 3.18 (m, 4H), 3.14 - 3.06 (m, 2H), 2.90 - 2.82 (m, 1H), 2.77 - 2.67 (m, 2H), 2.34 - 2.26 (m, 1H), 2.22 (p, J = 6.8 Hz, 2H), 2.18 - 2.08 (m, 3H), 1.64 (td, J = 15.0, 3.5 Hz, 2H). 49    3-((4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₀ H ₄₇ F ₄ N ₇ O ₃ : 749.86, found: 750.30. ¹ H NMR (600 MHz, MeOD) δ 8.16 (d, J = 1.1 Hz, 1H), 7.87 - 7.82 (m, 2H), 7.74 (s, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.49 (dd, J = 8.5, 1.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 2H), 7.06 (t, J = 9.1 Hz, 1H), 6.78 - 6.71 (m, 2H), 4.50 (s, 2H), 4.46 (t, J = 7.0 Hz, 2H), 4.40 (dd, J = 12.2, 5.0 Hz, 1H), 3.79 - 3.71 (m, 3.69 (s, 3H), 3.58 (d, J = 12.8 Hz, 2H), 3.49 - 3.42 (m, 2H), 3.32 - 3.26 (m, 3H), 3.21 (d, J = 6.9 Hz, 2H), 3.20 - 3.13 (m, 2H), 3.02 - 2.98 (m, 2H), 2.85 - 2.72 (m, 2H), 2.38 - 2.23 (m, 4H), 2.17 (d, J = 14.3 Hz, 2H), 1.75 - 1.65 (m, 2H). 50    5-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C43H47N6O5 : 784.36} ; found: 785.3. ¹ H NMR (600 MHz, DMSO ) δ 11.10 (s, 1H), 8.04 (br, 1H), 7.89 (s, 1H), 7.80 (d, J = 8.4 Hz, 2H), 7.67 (t, J = 8.4 Hz, 2H), 7.45 (d, J = 8.4 Hz , 2H), 7.32 (br, 2H), 7.25 (d, 1H), 5.06 (dd, J = 13.2 Hz, 5.4Hz, 1H), 4.38 (s, 1H), 4.26 (t, J = 7.2 Hz 2H), 2.91 (m, 3H), 2.61 (m, 2H), 2.47 (s, 3H), 2.17 (s, 3H), 1.84 (m, 4H), 1.28 (m, 7H), 1.18 (br, 2H), 0.91 (t, J = 7.2 Hz, 3H). 51    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₃₉ H ₃₉ F ₃ N ₆ O ₅ 728.77, found 729.20. ¹ H NMR (600 MHz, MeOD) δ 8.07 (s, 1H), 7.82 - 7.78 (m, 3H), 7.68 (s, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.50 (d, J = 2.3 Hz, 1H), 7.40 - 7.35 (m, 3H), 7.34 (dd, J = 8.4, 1.5 Hz, 1H), 5.12 (dd, J = 12.8, 5.5 Hz, 1H), 4.49 (s, 2H), 3.62 (d, J = 13.0 Hz, 2H), 3.58 - 3.44 (m, 3H), 3.20 (d, J = 6.9 Hz, 2H), 3.13 - 3.07 (m, 2H), 2.92 - 2.85 (m, 1H), 2.81 - 2.70 (m, 2H), 2.33 - 2.23 (m, 1H), 2.19 - 2.11 (m, 3H), 1.68 - 1.54 (m, 2H). 52    5-(4-((1-((1-((2,3-dihydrobenzo[b][1,4]dioxadien-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxapiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C48H47F3N6O7 : 876.93} , found: 877.3. ¹ H NMR (600 MHz, MeOD) δ 8.04 (d, J = 1.3 Hz, 1H), 7.79 - 7.75 (m, 2H), 7.69 - 7.62 (m, 3H), 7.38 - 7.32 (m, 4H), 7.21 (dd, J = 8.6, 2.3 Hz, 1H), 6.87 - 6.78 (m, 4H), 5.07 (dd, J = 12.8, 5.5 Hz, 1H), 4.62 - 4.59 (m, 4H), 4.40 (s, 2H), 4.36 - 4.32 (m, 1H), 3.96 (dd, J = 11.5, 5.7 Hz, 1H), 3.51 - 3.41 (m, 6H), 3.04 - 2.95 (m, 2H), 2.87 - 2.82 (m, 1H), 2.76 - 2.68 (m, 2H), 2.60 - 2.55 (m, 4H), 2.29 (d, J = 6.9 Hz, 2H), 2.12 - 2.02 (m, 3H), 1.91 (br, 1H), 1.43 (br, 2H). 53    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(pyridin-2-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C45H44F3N7O5 :} 819.34; found: 820. ¹ H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.53 (m, 1H), 8.01 (br, 2H), 7.81 (d, J =8.4 Hz, 2H), 7.76 (t, J =6.0 Hz, 1H), 7.67 (d, J =8.4 Hz, 1H), 7.62 (br, 1H), 7.46 (d, J =7.8 Hz 2H), 7.31 (m, 3H), 7.25 (d, J =8.4 Hz, 1H), 7.22 (d, J =8.4 Hz, 1H), 5.59 (s, 2H), 5.07 (dd, J =13.2 Hz, 5.4 Hz, 1H), 4.37 (br, 1H), 3.36 (m, 4H), 2.87 (m, 2H), 2.52 (m, 2H), 2.46 (br, 4H), 2.16 (br,2H), 2.02 (m, 2H), 1.89 (br, 2H), 1.76 (br, 2H), 1.23 (br, 4H). 54    3-((4-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M _] ⁺ calculated _{for C40H46F4N6O3 :} 734.84, found _: 735.40. ¹ H NMR (500 MHz, MeOD) δ 8.10 (d, J = 1.2 Hz, 1H), 7.82 - 7.76 (m, 2H), 7.71 (s, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.47 - 7.35 (m, 4H), 6.70 (ddd, J = 11.9, 11.5, 2.5 Hz, 2H), 4.51 - 4.35 (m, 5H), 3.75 - 3.66 (m, 2H), 3.66 - 3.52 (m, 4H), 3.09 - 2.95 (m, 4H), 2.88 - 2.70 (m, 2H), 2.34 - 2.23 (m, 3H), 2.14 - 1.94 (m, 5H), 1.86 - 1.74 (m, 2H), 1.67 - 1.49 (m, 4H). 55    3-((4-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidin-2,6-dione LC-MS (m/z): [ _{M ]} ⁺ calculated _{for C41H48F4N6O3 :} 748.37; found: 749.6. ¹ H NMR (600 MHz, d ⁴ -MeOD) δ 8.03 (s, 1H), 7.78 (d, 2H), 7.66 (s, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.36 (d, J = 7.8 Hz, 2H), 7.33 (m, 1H), 6.91 (t., J = 9.0 Hz, 1H), 6.55 (dd, J = 14.4 Hz, 2.4 Hz,1H), 6.50 (d, J = 8.4 Hz,1H), 4.35 (br, 2H), 4.30 (t, J = 7.2 Hz, 2H), 4.25 (dd, J = 12.0 Hz, 4.8 Hz, 1H), 3.49 (br, 3H), 2.98 (br, 6H), 2.80 (m, 1H), 2.74 (m, 1H), 2.62 (br, 4H), 2.32 (br, 3H), 2.04 (d, J = 12.9 Hz, 2H), 1.96 (s, 1H), 1.88 (m, 4H), 1.39 (m, 4H), 0.99 (t, J = 7.2 Hz 3H). 56    5-(4-((1-((1-((1,3-dioxolan-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxolan-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₃ H ₄₅ F ₃ N ₆ O ₇ 814.86, found 815.3 ¹ H NMR (600 MHz, MeOD) δ 8.02 (br, J = 1.3 Hz, 1H), 7.78 - 7.75 (m, 2H), 7.67 (dd, J = 8.5, 6.0 Hz, 2H), 7.63 (s, 1H), 7.38 - 7.32 (m, 4H), 7.22 (dd, J = 8.6, 2.3 Hz, 1H), 5.24 (t, J = 3.2 Hz, 1H), 5.07 (dd, J = 12.8, 5.5 Hz, 1H), 4.47 (d, J = 3.2 Hz, 2H), 4.41 (s, 2H), 3.83 - 3.78 (m, 2H), 3.76 - 3.71 (m, 2H), 3.54 - 3.41 (m, 6H), 3.06 - 2.98 (m, 2H), 2.88 - 2.81 (m, 1H), 2.76 - 2.68 (m, 2H), 2.62 - 2.55 (m, 4H), 2.30 (d, J = 6.4 Hz, 2H), 2.12 - 2.02 (m, 3H), 1.92 (s, 1H), 1.42 (br, 2H), 1.34 - 1.27 (m, 1H). 57    2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C46H52F3N5O5 :} 811.95; found: 813.3. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.06 (s, 1H), 7.79 (d, 2H), 7.69 (d, 1H), 7.67 (s, 1H), 7.62 (d, J =8.46 Hz, 1H), 7.37 (m, 4H), 7.24 (m, 1H), 5.09 (dd, J =13.2 Hz, 5.46 Hz, 1H), 4.43 (s, 2H), 4.29 (t, J =7.02 Hz, 2H), 3.46 (br, 4H), 3.03 (br, 2H), 2.87 (m, 1H), 2.74 (m, 2H), 2.61 (br, 4H), 2.32 (br, 2H), 2.09 (m, 3H), 1.99 (s, 1H), 1.92 (m, 3H), 1.60 (m, 1H), 1.32 (br, 3H), 1.26 (m, 2H), 0.91 (s, 3H), 0.90 (s, 3H). 58    3-((4-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl) -1H -indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₂ H ₅₁ F ₄ N ₇ O ₃ 777.91, found 778.50. ¹ H NMR (600 MHz, MeOD) δ 8.10 (d, J = 1.1 Hz, 1H), 7.83 - 7.77 (m, 2H), 7.73 (s, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.43 (dd, J = 8.5, 1.5 Hz, 1H), 7.39 (d, J = 8.0 Hz, 2H), 6.95 (t, J = 9.1 Hz, 1H), 6.61 - 6.51 (m, 2H), 4.49 (s, 2H), 4.44 (t, J = 7.0 Hz, 2H), 4.28 (dd, J = 11.9, 4.9 Hz, 1H), 3.79 - 3.63 (m, 3.57 (m, 2H), 3.85 (m, 2H), 3.59 (m, 2H), 3.43 - 3.36 (m, 2H), 3.33 - 3.25 (m, 2H), 3.25 - 3.13 (m, 6H), 3.10 (dd, J = 12.9, 10.8 Hz, 2H), 2.91 (s, 6H), 2.87 - 2.71 (m, 2H), 2.40 - 2.23 (m, 4H), 2.13 (d, J = 13.2 Hz, 2H), 1.97 (ddd, J = 25.4, 12.6, 4.8 Hz, 1H), 1.67 - 1.57 (m, 2H). 59    5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-dipiperidinyl]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₂ H ₄₅ F ₃ N ₆ O ₅ 770.85, found 771.3 ¹ H NMR (600 MHz, MeOD) δ 8.09 (d, J = 1.1 Hz, 1H), 7.82 - 7.78 (m, 2H), 7.69 (dd, J = 17.0, 8.3 Hz, 3H), 7.44 - 7.40 (m, 2H), 7.37 (d, J = 8.1 Hz, 2H), 7.29 (dd, J = 8.6, 2.2 Hz, 1H), 5.07 (dd, J = 12.8, 5.5 Hz, 1H), 4.44 - 4.39 (m, 4H), 4.03 (d, 3.52 (d, J = 12.7 Hz, 2H), 3.03 - 2.95 (m, 6H), 2.88 - 2.82 (m, 1H), 2.77 - 2.68 (m, 2H), 2.28 - 2.22 (m, 2H), 2.12 - 2.07 (m, 1H), 2.05 - 2.00 (m, 2H), 1.87 (d, J = 12.1 Hz, 2H), 1.57 - 1.45 (m, 4H), 1.43 - 1.35 (m, 2H). 60    3-((4-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₂ H ₅₀ F ₄ N ₆ O ₃ 762.89, found 763.4 ¹ H NMR (600 MHz, MeOD) δ 8.06 (d, J = 1.1 Hz, 1H), 7.81 - 7.76 (m, 2H), 7.70 (s, 1H), 7.65 (d, J = 8.5 Hz, 1H), 7.40 - 7.34 (m, 3H), 6.91 (t, J = 9.2 Hz, 1H), 6.54 - 6.45 (m, 2H), 4.41 - 4.35 (m, 4H), 4.22 (dd, J = 11.8, 4.9 Hz, 1H), 3.48 (d, J = 11.8, 4.9 Hz, 1H), 3.80 (d, J = 13.7, 13.9 Hz, 1H), 3.91 (d, J = 13.8, 13.9 Hz, 1H), 3H), 2.57 (t, J = 11.8 Hz, 2H), 2.32 - 2.24 (m, 3H), 2.00 (d, J = 12.9 Hz, 2H) , 1.86 (d, J = 12.6 Hz, 2H), 1.53 - 1.40 (m, 5H), 1.32 - 1.22 (m, 3H). 61    4-amino-N-(3-(5-((4-((4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidin-4-carboxamide LC-MS (m/z): [M] ⁺ calcd. for C ₄₆ H ₅₇ F ₄ N ₉ O ₄ 876.01, found 876.5 ¹ H NMR (600 MHz, MeOD) δ 8.12 (d, J = 1.1 Hz, 1H), 7.85 - 7.79 (m, 3H), 7.67 (d, J = 8.5 Hz, 1H), 7.43 (dd, J = 8.5, 1.5 Hz, 1H), 7.36 (d, J = 8.0 Hz, 2H), 7.10 (t, J = 9.0 Hz, 1H), 6.90 - 6.81 (m, 2H), 4.50 - 4.43 (m, 3H), 4.38 (t, J = 6.9 Hz, 3.76 - 3.68 (m, 2H), 3.56 (d, J = 13.2 Hz, 2H), 3.51 - 3.41 (m, 4H), 3.39 - 3.34 (m, 5H), 3.19 (d, J = 6.8 Hz, 2H), 3.16 - 3.08 (m, 2H), 2.79 - 2.67 (m, 4H), 2.34 - 2.26 (m, 1H), 2.23 - 2.11 (m, 7H), 2.03 - 1.95 (m, 3H), 1.72 - 1.62 (m, 2H), 1.41 - 1.34 (m, 2H). 62    5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-inden-5-yl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-N,N-dimethyl-3-(4-(trifluoromethoxy)phenyl)-1H-indole-1-sulfonamide LC-MS (m/z): [ _M ] ⁺ _{calcd .} for _{C42H45F3N6O7S :} 834.91; found: 836.3. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.12 (d, J =8.4 Hz, 1H), 8.00 (s, 1H), 7.90 (s, 1H), 7.83 (m, 2H), 7.70 (d, J =8.4 Hz, 1H), 7.50 (dd, J =7.2 Hz, 1.2 Hz, 1H) , 7.45 (d, J =7.8 Hz,2H), 7.24 (dd, J = 9.0 Hz, 2.4 Hz, 1H), 5.09 (dd, J =12.6 Hz, 5.4 Hz,, 1H), 4.63 (br, 1H), 4.25 (br, 1H), 3.46 (t, J =5.4 Hz,, 4H), 3.36 (m, 2H), 2.86 (m, 1H), 2.76 (m, 4H), 2.61 (t, J =4.8Hz, 4H), 2.32 (d, J =7.2 Hz, 2H), 2.12 (m, 1H), 2.01 (d, J =14.4 Hz, 2H), 1.98 (s, 1H), 1.87 (br, 1H), 1.41 (m, 2H), 1.32 (m, 4H). 63    3-((3-Fluoro-4-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)phenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ _{calcd . for C43H52F4N6O3 :} 776.24; found: 777.5. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.05 (s, 1H), 7.79 (d, J = 6.6 Hz, 2H), 7.67 (s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.36 (m, 3H), 6.90 (t, J =9.0Hz, 1H), 6.55 (dd, J = 14.4 Hz, 2.58 Hz, 1H), 6.50 (d, J = 8.64 Hz, 6.06 Hz, 1H), 4.41 (br, 2H), 4.29 (t, J =7.0 Hz, 2H), 4.25 (dd, J = 11.8 Hz, 4.86 Hz, 1H), 3.50 (m, 7 (m, 2H), 1.54 (m, 4H), 1.63 (m, 1H), 1.29 (m, 2H), 0.76 (s, 3H), 0.91 (s, 3H), 0.90 (s, 3H). 64    3-((4-fluoro-3-(4-((1-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)phenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ calculated for C ₃₇ H ₄₀ F ₄ N ₆ O ₃ : 692.31: Found: 693.6. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.05 (s, 1H), 7.80 (d, J =8.4 Hz, 1H), 7.66 (s, 1H), 7.58 (d, 1H), 7.37 (d, J =8.4 Hz, 2H), 7.31 (d, J =7.8 Hz, 1H), 6.91 (t, J =9.6 Hz, 1H), 6.55 (dd, J =13.08 Hz, 2.4 Hz, 1H), 6.50 (d, J =8.4 Hz, 1H), 4.42 (s, 2H), 4.24 (dd, J = 11.4 Hz, 4.8 Hz, 1H), 3.50 (br, 2H), 3.0 δ 1.14 (m, 2H), 1.43 (br, 2H), 1.31 (br, 2H). 65    3-((3-Fluoro-4-(1'-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)phenyl)amino)piperidine-2,6-dione LC-MS (m/z): [ _{M]+ calculated for C37H39F4N5O3 :} ^677.30 _{; found :} 678.2. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.05 (d, J = 0.6 Hz, 1H), 7.80 (d, J = 6.6 Hz, 2H),7.66 (s, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.37 (d, J = 7.8 Hz, 2H), 7.31 (dd, J = 8.4 Hz, 1.2 Hz, 1H), 6.92 (t, J = 9.0 Hz, 1H), 6.54 (dd, J = 14.4 Hz, 2.4 Hz, 1H), 6.49 (dd, J = 8.4 Hz, 2.4 Hz 1H), 4.04 (s, 2H), 4.24 (dd, J = 12.0 Hz, 4.8 Hz 1H), 3.52 (d, J = 12 Hz, 2H), 3.27 (br, 2H), 3.00 (br, 2H), 2.85-2.77 (m, 1H), 2.72 (m, 1H), 2.59 (t, J = 12 Hz, 2H), 2.31 (m, 1H), 2.04 (br, 2H), 1.98 (s, 2H), 1.93 (m, 1H), 1.81 (br, 2H), 1.56-1.42 (m, 5H), 1.33-1.22 (m, 2H). 66    5-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₃₈ H ₃₇ F ₃ N ₆ O ₅ 714.75, found 715.1 ¹ H NMR (600 MHz, MeOD) δ 8.07 (br, 1H), 7.79 - 7.75 (m, 2H), 7.68 (s, 1H), 7.65 (dd, J = 8.4, 2.9 Hz, 2H), 7.43 (dd, J = 8.5, 1.2 Hz, 1H), 7.34 (d, J = 8.1 Hz, 2H), 7.05 (d, J = 2.1 Hz, 1H), 6.93 (dd, J = 8.5, 2.1 Hz, 1H), 5.07 (dd, J = 12.8, 5.5 Hz, 1H), 4.62 (s, 4H), 4.46 - 4.37 (m, 4H), 3.66 - 3.48 (m, 6H), 3.19 (s, 2H), 3.00 - 2.94 (m, 2H), 2.89 - 2.82 (m, 1H), 2.77 - 2.67 (m, 2H), 2.28 - 2.21 (m, 2H), 2.13 - 2.07 (m, 1H). 67    3-((4-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-fluorophenyl)amino)piperidine-2,6-dione LC-MS (m/z): [M] ⁺ calculated _{for C36H38F4N6O3 : 678.73} , found _: 679.2. ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 10.81 (s, 1H), 8.21 - 8.15 (m, 1H), 8.02 - 7.93 (m, 4H), 7.85 (dd, J = 8.9, 2.3 Hz, 2H), 7.73 (dd, J = 12.5, 8.5 Hz, 1H), 7.52 - 7.42 (m, 3H), 6.78 (s, 1H), 6.58 (d, J = 15.5 Hz, 1H), 6.46 (s, 1H), 4.52 (d, J = 5.6 Hz, 2H), 4.44 (s, 1H), 4.39 (d, J = 6.8 Hz, 2H), 4.28 (d, J = 11.1 Hz, 1H), 3.21 (s, 2H), 2.93 (d, J = 31.9 Hz, 4H), 2.80 (m, 4H), 2.15 - 2.07 (m, 3H), 1.90 - 1.82 (m, 1H). 68    5-(4-((1-((1-(3,3-dimethoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ _{calcd . for C44H49F3N6O7 :} 830.36; found: 831.4. ¹ H NMR (600 MHz, d ⁴ -MeOD ) δ 8.04 (s, 1H), 7.79 (d, J = 9.0 Hz, 2H), 7.69 (d, J = 8.4 Hz, 1H), 7.66 (s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.37 (m, 4H), 7.24 (dd, J = 8.4 Hz, 2.4 Hz, 1H), 5.09 (dd, J = 12.6 Hz, 5.4 Hz, 1H), 4.63 (s, 1H), 4.38-4.31 (m, 5H), 3.46 (br, 6H), 3.34 (st, 6H), 2.78 (m, 1H), 2.74 (m, 1H), 2.60 (br, 4H), 2.32 (d, J =6.6Hz, 1H), 2.17 (m, 2H), 2.12 (m, 1H), 2.05 (br, 2H), 1.96 (s, 1H), 1.32 (br, 4H). 69    5-(5-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₀ H ₄₁ F ₃ N ₆ O ₅ 742.80, found 743.2 ¹ H NMR (600 MHz, MeOD) δ 7.99 (s, 1H), 7.76 - 7.72 (m, 2H), 7.65 (d, J = 8.6 Hz, 2H), 7.58 (d, J = 8.5 Hz, 1H), 7.35 (dd, J = 8.5, 1.5 Hz, 1H), 7.31 (d, J = 8.1 Hz, 2H), 7.03 (d, J = 2.2 Hz, 1H), 6.91 (dd, J = 8.5, 2.2 Hz, 1H), 5.09 (dd, J = 12.8, 5.5 Hz, 1H), 4.36 (t, J = 6.9 Hz, 2H), 4.25 (s, 2H), 3.55 - 3.48 (m, 4H), 3.47 - 3.41 (m, 2H), 3.22 (s, 2H), 3.05 - 2.95 (m, 4H), 2.87 - 2.82 (m, 1H), 2.77 - 2.74 (m, 1H), 2.72 (s, 6H), 2.29 - 2.23 (m, 2H), 2.12 - 2.07 (m, 1H). 70    5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[3,3'-azocyclobutane]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₃₈ H ₃₇ F ₃ N ₆ O ₅ 714.75, found 715.1 ¹ H NMR (600 MHz, MeOD) δ 8.00 (s, 1H), 7.76 (dd, J = 8.8, 2.1 Hz, 2H), 7.69 - 7.68 (m, 1H), 7.58 (d, J = 8.3 Hz, 1H), 7.38 - 7.34 (m, 3H), 7.30 (ddd, J = 8.4, 5.5, 1.5 Hz, 1H), 7.01 (t, J = 1.9 Hz, 1H), 6.87 (dd, J = 8.4, 2.1 Hz, 1H), 5.03 (ddd, J = 8. = 12.8, 5.5, 2.4 Hz, 1H), 4.43 - 4.39 (m, 4H), 4.22 - 4.12 (m, 3H), 4.07 - 4.02 (m, 1H), 3.71 - 3.67 (m, 2H), 2.99 - 2.93 (m, 3H), 2.88 - 2.83 (m, 2H), 2.76 - 2.68 (m, 2H), 2.56 - 2.50 (m, 1H), 2.26 - 2.20 (m, 3H), 2.11 - 2.07 (m, 1H). 71    5-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-dipiperidinyl]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione LC-MS (m/z): [M] ⁺ calcd. for C ₄₄ H ₄₉ F ₃ N ₆ O ₅ 798.91, found 799.4 ¹ H NMR (600 MHz, MeOD) δ 8.07 - 8.04 (m, 1H), 7.79 - 7.75 (m, 2H), 7.70 (s, 1H), 7.65 (dd, J = 8.5, 5.7 Hz, 2H), 7.40 - 7.34 (m, 3H), 7.32 (d, J = 2.3 Hz, 1H), 7.19 (dd, J = 8.7, 2.3 Hz, 1H), 5.06 (dd, J = 12.8, 5.5 Hz, 1H), 4.43 - 4.36 (m, 4H), 3.07 (d, J = 13.3 Hz, 2H), 3.56 (d, J = 11.0 Hz, 2H), 3.14 - 3.08 (m, 2H), 3.00 - 2.90 (m, 4H), 2.90 - 2.83 (m, 1H), 2.81 (s, 6H), 2.77 - 2.68 (m, 2H), 2.34 - 2.27 (m, 2H), 2.12 - 2.07 (m, 1H), 2.01 (d, J = 12.0 Hz, 2H), 1.85 (d, J = 12.3 Hz, 2H), 1.46 (s, 4H), 1.37 - 1.29 (m, 2H). 72    (2R,4S)-1-((R)-2-(2-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)acetamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((R)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide LC-MS (m/z): [M] ⁺ calcd. for C ₄₄ H ₄₉ F ₃ N ₆ O ₅ 1013.52, found 1014.7 ¹ H NMR (600 MHz, DMSO-d6) δ 9.67 (s, 1H), 9.00 (s, 1H), 8.45 - 8.38 (m, 1H), 8.06 (s, 1H), 7.91 (d, J = 2.6 Hz, 1H), 7.88 - 7.75 (m, 5H), 7.71 (dd, J = 8.3, 3.8 Hz, 1H), 7.49 - 7.33 (m, 6H), 4.92 - 4.89 (m, 1H), 4.53 (d, J = 9.2 Hz, 1H), 3H), 4.41 (t, J = 6.6 Hz, 3H), 4.36 (t, J = 7.1 Hz, 2H), 4.30 (s, 2H), 3.97 (s, 2H), 3.64 - 3.61 (m, 2H), 3.55 (d, J = 10.8 Hz, 2H), 3.43 (s, 2H), 2.94 - 2.86 (m, 3H), 2.83 (d, J = 7.2 Hz, 2H), 2.46 (s, 3H), 2.10 (d, J = 7.7 Hz, 2H), 1.92 (d, J = 13.4 Hz, 2H), 1.79 (m, 1H), 1.38 (d, J = 7.0 Hz, 4H), 0.95 (s, 9H).

Examples 6 Biological Activity

Various RAS-PROTACs were tested for their specificity and ability to degrade the target protein. A brief description of the different assays is described below.

Western blotting to evaluate the cellular efficacy of RAS-PROTAC in KRAS protein degradation

In Western blot experiments, BT-474 (KRAS ^WT ) and MIA PaCa-2 (KRAS ^G12C ) cells were cultured in DMEM medium with 10% FBS. BxPC3 (KRAS ^WT ) and KLM-1 (KRAS ^G12D ) cells were cultured in DMEM medium with 10% FBS. MDA-MB-231 (KRAS ^G13D ) cells were cultured in L15 medium with 10% FBS. BT-474, MIA PaCa-2, BxPC3, and MDA-MB-231 have been established by the American Type Culture Collection. KLM-1 has been established by ExPASy. On the day of analysis, 200,000 cells were pretreated with each test compound for 24 hours. After 24 hours, whole cell lysates were collected by adding 2× SDS sample buffer. Proteins were separated by SDS-PAGE electrophoresis and transferred to PVDF membranes. Protein expression was detected using immunoblotting with various primary and secondary antibodies following standard protocols. Antibodies against KRAS were purchased from Abcam (Cambridge, UK). Anti-rabbit IgG, HRP-linked secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). Antibodies against actin were purchased from Millipore (Burlington, MA). Immunoblots were visualized by chemiluminescence (SuperSignal™ West Femto Maximum Sensitivity Substrate, Thermo Fisher, Waltham, MA) and detected by ChemiDoc ^TM MP Imaging System (Bio-Rad, Hercules, CA). The intensity of the bands in the Western blots was also quantified by the ChemiDoc ^™ MP imaging system. The relative intensity of the bands corresponding to the drug-treated groups was compared with that of the untreated groups.

The preliminary screening results of bifunctional compounds 1-21 for degradation of KRAS protein in Calu-1, H358, H441, KLM-1, MDA-MB-231 and HCT-116 cell lines are shown in Table 2, which were tested under the conditions described above.

Table 2 DC50 G12C G12V G12D G13D G12S cpd number MW (Da) Calu-1 H358 H441 KLM-1 MDA-MB-231 HCT-116 A549 1 876.98 ++ + + ++ 2 1003.14 - - - - - 3 785.87 ++ + ++ ++ ++ ++ 4 717.87 - - - - - 5 717.87 - - - - - 6 785.87 ++ + ++ ++ ++ ++ 7 719.86 ++ - + + + + 8 688.71 + + + + + 9 698.18 ++ + + + + 10 731.9 + + + + + + 11 726.88 - - - - - - 12 732.89 - - - - - 13 726.88 - + - - + 14 785.87 + + + + + + 15 749.89 + + + + + 16 785.87 + + + + + 17 732.89 - - - - - - 18 769.87 + + + + + + 19 785.87 + + + + + + 20 785.87 ++ + Not Measured Not Measured Not Measured Not Measured twenty one 885.99 + + + - - + twenty two 741.77 + + + + + + 30 785.87 +++ ++ ++ ++ ++ ++ +++ 37 813.92 ++ ++ ++ ++ ++ ++ ++ 49 749.86 ++ ++ ++ ++ ++ ++ ++ 54 734.84 +++ +++ +++ +++ +++ +++ +++ 59 770.85 ++ ++ ++ ++ ++ ++ ++ “ND” means not determined, “++++” ＜ 0.3 µM, 0.3 µM ≤ “+++” ＜ 1.0 µM, 1.0 µM ≤ “++” ＜ 3.0 µM, 3 µM ≤ “+” ＜ 10.0 µM, “━” means ≥ 10 µM.

Examples 7 Antiproliferative activity

As mentioned above, the KRAS-PROTAC of the present invention can be used to treat diseases or disorders with specific KRAS mutations. Such diseases may be cancer, autoimmune diseases, infectious diseases, or angiogenic disorders. The cancer may be lung cancer (e.g., non-small cell lung cancer), colorectal cancer, colorectal cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, bladder cancer, gastric cancer, kidney cancer, salivary gland cancer, ovarian cancer, uterine cancer, cervical cancer, oral cancer, skin cancer, brain cancer, lymphoma, leukemia, bile duct malignancy, endometrial cancer, or myeloid leukemia. The KRAS-PROTAC ^of the present invention was used to measure the inhibition of cell growth. The cytotoxicity of KRAS-PROTAC was evaluated in lung cancer cell lines, breast cancer cell lines, and pancreatic cancer cell lines with different RAS mutations. The results are shown in Table 3.

table 3 IC ₅₀ (µM) G12C G12V G12D G13D G12S cpd number Calu-1 H358 H441 KLM-1 MDA-MB-231 HCT-116 A549 1 ++ + ++ ++ ++ 2 - - - - - 3 ++ ++ + ++ + + 4 - - - - - - 5 - - - - - - 6 ++ ++ + ++ + ++ 7 + + + + + + 8 + + + + + + 9 ++ ++ ++ ++ ++ ++ 10 ++ + + ++ + ++ 11 + + + + + + 12 + - - - + + 13 + + + + + + 14 ++ + + + + ++ 15 ++ + + + + + 16 ++ + + + + ++ 17 - - - - - - 18 ++ + + + + + 19 ++ + + + + + 20 ++ + + ++ ++ ++ twenty one ++ + + + + ++ twenty two ++ + + ++ + ++ twenty three + + - + + + twenty four + - - + + + 25 ++ ++ + ++ ++ ++ 26 ++ + + + + ++ 27 - - - + + + 28 ++ ++ + ++ + ++ 29 + + + + + + 30 ++ ++ ++ ++ ++ ++ +++ 31 + + + ++ ++ ++ 32 ++ ++ + ++ + ++ 33 ++ ++ ++ ++ ++ +++ 34 ++ + + ++ + ++ 35 + + - + + + 36 + + + + + + 37 ++ ++ ++ ++ ++ +++ 38 + ++ + ++ + ++ 39 ++ ++ + ++ + ++ 40 + + + - 41 + + + + 42 + + + - 43 + + ++ + 44 + + + + 45 + + + - 46 + + + + 47 + - + + 48 + - + + 49 ++ ++ + ++ ++ +++ ++ 50 ++ + ++ + 51 ++ + + ++ 52 ++ - + ++ 53 + + + + 54 +++ ++ ++++ ++++ ++ ++ ++++ 55 ++ + ++ + 56 + + + + 57 ++ + ++ + 58 ++ + ++ ++ 59 ++ ++ ++ ++ ++ ++ ++ 60 ++ + ++ ++ 61 + + + + 62 + - + + 63 ++ + ++ ++ 64 ++ + ++ ++ 65 ++ + ++ ++ 66 ++ + ++ ++ 67 ++ + ++ + 68 ++ + + + 69 +++ + ++ + 70 - - + + 71 ++ + ++ ++ 72 + ++ + ++ ++ ++ “ND” means not determined, “++++” ＜ 0.3 µM, 0.3 µM ≤ “+++” ＜ 1.0 µM, 1.0 µM ≤ “++” ＜ 3.0 µM, 3 µM ≤ “+” ＜ 10.0 µM, “━” means ≥ 10 µM.

Examples 8 Ras-PROTAC Xenotransplantation Model ( Lung cancer )

The goal of this study was to evaluate the in vivo antitumor efficacy of Ras-PROTAC in the Calu-1 human lung cancer xenograft model in male NOD SCID mice.

The test article Cpd 6 and the corresponding vehicle were formulated and injected intratumorally (IT) to mice once a day for 14 consecutive days. Calu-1 cells were maintained in vitro as monolayer cultures in RPMI-1640 medium supplemented with 10% fetal bovine serum at 37°C in an air atmosphere containing 5% _CO2 . Tumor cells were subcultured in the conventional manner by trypsin-EDTA treatment twice a week. Growing cells in the exponential growth phase were collected and counted for tumor inoculation. Male NOD SCID mice aged 6-7 weeks were purchased from BioLasco Taiwan and isolated for one week. Six mice were housed in each cage. All animals were housed in an animal facility at 19-25°C with a 12-h light/12-h dark cycle. Animals had free access to rodent pelleted food and water ad libitum. Calu-1 cells were implanted subcutaneously (SC) into the right flank of male NOD SCID mice (5×10 ⁶ cells in a 1:1 PBS/Matrigel mixture, 0.1 mL per mouse). When the mean tumor volume reached 170 mm ³ , mice were randomized into 2 groups (N=6 per group). Vehicle or Cpd 6 (10 mg/kg) was given by intratumoral (IT) injection once daily for 14 consecutive days. Tumor volume, body weight, mortality, and overt signs of toxicity were monitored and recorded three times a week for 28 days. Tumor volume (mm ³ ) was measured three times per week using a caliper and calculated according to the following formula: Tumor volume = ( w ² × l )/2, where w = tumor width and l = tumor diameter length (mm). The percentage of tumor growth inhibition (TGI) was calculated using the following formula: %TGI = [1 - (T/C)] × 100%, where T and C represent the average tumor volume of the treated and control groups, respectively. The results are shown in Table 4 and Figures 1 and 2. The Student's t test was used to compare the vehicle-treated group and the test substance-treated group. *Differences were considered significant when P < 0.05. The animals were weighed three times per week until the completion of the study. Weight change was calculated as the percentage of weight gain compared to the initial weight.

Table 4 Calu-1 xenograft model treatment TGI % (1-T/C) D0 D3 D5 D7 D10 D12 D14 D17 D19 D21 D24 D26 D28 Media, IT, QD×14 0 0 0 0 0 0 0 0 0 0 0 0 0 Cpd6, 10 mpk, IT, QD×14 0 56 80 82 90 93 91 93 93 93 89 87 86

Figure 1 shows the tumor growth curves of Calu-1 implanted male NOD SCID mice. The test article Cpd 6 at 10 mg/kg significantly reduced Calu-1 tumor growth from day 3 to day 28, with TGI values ranging from 56% to 93%, and is considered very effective against Calu-1 xenografts.

Figure 2 shows the body weight changes of male NOD SCID mice implanted with Calu-1. No weight loss was observed during the entire experiment.

For cancer patients with KRAS mutations, the KRAS-PROTAC of the present invention is a promising new therapeutic agent.

This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any incorporated reference and this specification, this specification shall prevail. In addition, specific embodiments of the invention that are prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are considered to be known to those of ordinary skill in the art, they may be excluded even if they are not explicitly stated to be excluded herein. Any specific embodiment of the invention may be excluded from any claim for any reason, whether or not related to the existence of prior art.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. The scope of the embodiments of the invention described herein is not intended to be limited to the above description, but is actually described in the appended claims. Those skilled in the art will appreciate that various changes and modifications may be made to this specification without departing from the spirit or scope of the invention as defined by the following claims.

without

Figure 1 shows the tumor growth curves of Calu-1-implanted male NOD SCID mice.

Figure 2 shows the weight changes of male NOD SCID mice implanted with Calu-1.

Claims (22)

A bifunctional compound of formula (I): RB-Linker-ULM (I) or a pharmaceutically acceptable salt, hydrate, tautomer or stereoisomer thereof, wherein: ULM is an E3 ubiquitin ligase binding moiety or a chaperone protein complex binding moiety; the linker is a group covalently bonded to the RB and ULM moieties; and RB is a RAS protein binding moiety, and is represented by the formula RB-I:

wherein: _X1 is H, ^NRX1RX2 , ^SO2NRX1RX2 , _ORX1 , ^CHRX1RX2 , NH(C=O) ^RX3 or CN; ^RX1 and ^RX2 are each independently H, OH or a linear or branched _C1-4 alkyl group, which ^is optionally substituted by ^one or ^more halogen groups ^{, or RX1 and RX2} together with the atoms to which they are attached form a 4- to 8-membered ring containing 0 to 2 heteroatoms and which is optionally substituted by one or more _C1-4 alkyl groups; ^RX3 is a linear or branched _C1-10 alkyl group, which is optionally substituted by one or more halogen groups, or a linear or branched C1-10 alkyl group, which is optionally substituted by one or more halogen groups. _W1 is a bond, _C1-6 alkyl, alicyclic, heterocyclic, bicyclic or biheterocyclic, each of which is optionally substituted by one, _two or three _RW1 , and each ^RW1 is independently H, halogen, OH, _NH2 , _NMe2 , _NEt2 , CN, _C1-4 alkyl optionally substituted by one or more F, or _C1-3 alkoxy optionally substituted by one or more F; ^R1 is CH, C-Me, C _- halogen or N _;

is an aryl, heteroaryl or indazole, which is independently substituted by one or more halogen groups, CF ₃ , OCF ₃ , OR ^{A1 -RA2} , hydroxyl, nitro, CN, C≡CH, a straight or branched C _1-6 alkyl group optionally substituted by one or more halogen groups or C 1-6 alkoxy groups, a straight or branched C _1-6 alkoxy group optionally substituted by one or more halogen groups, a C _2-6 alkenyl group, _{a C 2-6 alkynyl} group or a 4- to 7-membered saturated or partially unsaturated heterocyclic ring containing 1-2 heteroatoms, ^RA1 is a C _1-6 alkyl group, ^RA2 is a heteroaryl group containing 1 to 2 heteroatoms and optionally substituted by one or more halogen groups; X ₂ is CH ₂ or C═O;

is a 4- to 8-membered alicyclic ring having 0 to 4 heteroatoms, which is optionally substituted with 0 to 4 R ^Q , each R ^Q is independently OH, or a straight or branched C _1-6 alkyl group optionally substituted with one or more halogen groups, or two R ^Q groups together with the atoms to which they are attached form a 3- to 8-membered ring containing 0 to 2 heteroatoms; Z ₁ is H, C _1-4 alkyl, C _1-4 alkoxy, halogen, OH, CN or C _2-4 alkynyl; and the dotted line indicates the point of attachment to the linker; wherein ULM is Cereblon E3 ubiquitin ligase binding moiety (CLM), Von Hippel-Lindau (Von Hippel-Lindau) Hippel-Lindau (VHL) E3 ubiquitin ligase binding portion (VLM), DDB1-related and CUL4-related factor 16 (DCAF16) E3 ubiquitin ligase binding portion (DLM), IAP E3 ubiquitin ligase binding portion (ILM), mouse two microsome 2 (MDM2) homolog E3 ubiquitin ligase binding portion (MLM), Kelch-like ECH-associated protein-1 (KEAP1) E3 ubiquitin ligase binding portion, DCAF15 E3 ubiquitin ligase binding portion, RNF4 E3 ubiquitin ligase binding portion, RNF114 E3 ubiquitin ligase binding portion or aromatic hydrocarbon receptor (AhR) E3 ubiquitin ligase binding portion. The bifunctional compound of claim 1, wherein the ULM is a CLM having a chemical structure represented by CLM-a or CLM-b:

wherein: W is selected from CH ₂ and C=O; and Q ₁ , Q ₂ , Q ₃ and Q ₄ are each independently C, N or C-halogen, and wherein one of Q ₁ , Q ₂ , Q ₃ and Q ₄ is covalently linked to a linker;

The dotted lines represent the connection points with the connectors. The bifunctional compound of claim 2, wherein the ULM is a CLM represented by:

Wherein: the dotted line indicates the connection point with the linker; and X _CLM is a halogen. The bifunctional compound of claim 1, wherein the ULM is a VLM having a chemical structure represented by one of the following chemical structures:

,

wherein: R ₅ is H or a linear or branched C _1-3 alkyl group; R ₆ is CN or a 5-membered heteroaryl group having one or two heteroatoms selected from N, S or O, which is optionally substituted with a methyl group; Z ₂ is F or CN; and the dotted line indicates the point of connection to the linker. The bifunctional compound of claim 1, wherein ULM is DLM having a chemical structure represented by D16-a:

wherein: the dashed line connected to Y ₁ indicates the point of connection to the linker; R ₇ is H, halogen, C _1-4 alkyl or C _1-4 alkoxy; Y ₁ is O or S or NH; and R ^m is a covalent electrophile selected from

The bifunctional compound of claim 1, wherein the ULM is a DLM represented by:

The dotted lines represent the connection points with the connectors. The bifunctional compound of claim 1, wherein the RB-I is selected from the group consisting of formula RB-1, RB-3, and RB-5 to RB-7:

The dotted lines represent the connection points with the connectors. The bifunctional compound of claim 1, wherein the RB-I is of formula RB-2:

The dotted lines represent the connection points with the connectors. The bifunctional compound of claim 1, wherein the RB-I is represented by:

The dotted lines represent the connection points with the connectors. The bifunctional compound of claim 1, wherein the RB-I is selected from the group consisting of:

The dotted lines represent the connection points with the connectors. The bifunctional compound of any one of claims 1 to 5, which is selected from Formula Ia and Ic to Formula Ig:

The bifunctional compound of any one of claims 1 to 5, which is of formula Ib:

The bifunctional compound of claim 1, wherein the linker comprises the following chemical structure:

Wherein: the dotted line of the linker is the point of connection with RB or ULM; Y ^L2 is a bond, or a straight or branched chain C _1-4 alkyl or alkoxy group optionally substituted with halogen, C _1-3 alkyl, methyl or ethyl; W ^L3 is a bond, -C=OCH ₂ -, an aryl group, or a 3- to 7-membered ring or an 8- to 12-membered spiro ring, each containing 0 to 4 heteroatoms and optionally substituted with halogen or methyl; Y ^L3 is a bond or a C _1-32 alkyl alkenyl or alkynyl group, wherein one or more C atoms are optionally substituted with O,

or NH, and each carbon is optionally substituted by halogen, =0, methyl or ethyl, and each nitrogen is optionally substituted by methyl or ethyl; Y ^L4 is a bond, O or unsubstituted or substituted straight or branched C _1-6 alkyl, wherein one or more carbons are optionally replaced by O, NH or NCH ₃ , and optionally substituted by halogen or methyl; W ^L4 is a 3- to 8-membered ring or a 5- to 8-membered spiro ring, each having 0 to 4 heteroatoms and optionally substituted by halogen or methyl; and Y ^L5 is a bond or C _1-6 alkyl, wherein one or more C atoms are optionally replaced by O and optionally substituted by halogen or methyl. The bifunctional compound of claim 13, wherein the linker has a chemical structure represented by:

The dotted line of the connector is the connection point with RB or ULM. A bifunctional compound selected from the group consisting of compounds (cpd) 1-72: 5-(4-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin

-1-yl)methyl)benzyl)piperidin

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 1); 4-amino-N-(3-(5-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

-1-yl)methyl)benzyl)piperidin

-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidine-4-carboxamide (cpd 2); 5-(4-((4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-carboxamide

-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 3); 5-(4-((4-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 4); 5-(4-((1-((1-(3-aminopropyl)-3-(4-hydroxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 5); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 6); 5-(4-((4-((1-(3-aminopropyl)-3-(4-fluorophenyl)-1H-indol-5-yl)methyl)piperidin-1,3-dione

-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 7); 5-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 8); 1-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1,3-dione

-1-yl)-2-((1-(2-chloroacetyl)-1,2,3,4-tetrahydroquinolin-6-yl)oxy)ethan-1-one (cpd 9); 5-(4-((4-((1-(3-aminopropyl)-3-(4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-1-one

-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 10); 4-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 10);

5-(4-((1-((1-(3-aminopropyl)-3-(6-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)benzonitrile (cpd 11);

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 12); 3-(1-(3-aminopropyl)-5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-7-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)methyl)-1H-indol-3-yl)benzonitrile (cpd 13); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-7-yl)methyl)piperidin-4-yl)methyl)piperidin-1 ...

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 14); 5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-methoxyphenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 15); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-6-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 16); 5-(4-((1-((1-(3-aminopropyl)-3-(2-methoxypyridin-3-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 17); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethyl)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 18); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-4-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 19); 4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (cpd 20); (3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)carbamic acid tributyl ester (cpd 21); 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)prop-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (cpd 22); 5-(3-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)propyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (Cpd 23); 5-(4-(2-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)oxy)propyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

-1-yl)acetyl)piperidin

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (cpd 24); 5-(4-((1-((1-(3-aminopropyl)-3-(3-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 25); 5-(4-((1-(1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indole-5-carbonyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 26); 5-(4-((1-((1-(3-aminopropyl)-3-(3-hydroxynaphthalen-1-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 27); 5-(4-((1-((1-(azacyclobutan-3-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 28); 5-(4-((1-((1-(3-aminopropyl)-3-(5-chloro-6-methyl-1H-indazol-4-yl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 29); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 30); (2S)-N-(2-(3-(4-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-1,3-dione

-1-yl)propyloxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carboxamide (Cpd 31); 5-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-

-1-yl)-2-(2,6-dioxopiperidin-3-yl)-6-fluoroisoindole-1,3-dione (Cpd 32); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(methylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-

-1-yl)isoindoleline-1,3-dione (Cpd 33); 5-(7-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoleline-1,3-dione (Cpd 34); 5-(4-((1-((1-(3-aminopropyl)-3-(2-fluoro-4-((3-fluoropyridin-2-yl)methoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-5-yl

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 35); 5-(4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)azepanobutyl-3-yl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 36); 5-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 37); 5-(4-((1-((1-(5-aminopentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (Cpd 38); 5-(7-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)-2,7-diazaspiro[3.5]nonan-2-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (Cpd 39); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)acetamide (Cpd 40); 4-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)acetamide (Cpd 40);

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)butyronitrile (Cpd 41); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-methyl-1H-imidazol-5-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-5-yl)methyl)piperidin-1-yl

-1-yl)isoindole-1,3-dione (Cpd 42); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-methoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)isoindole-1,3-dione (Cpd 43); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(3-(2,2,2-trifluoroethoxy)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-5-yl

-1-yl)isoindolin-1,3-dione (Cpd 44); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-1H-pyrrole-2-carboxamide (Cpd 45); 3-((4-(1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 46); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-3-(trifluoromethyl)benzamide (Cpd 47); N-(3-(5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl ...

3-((4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1-yl)methyl)piperidin-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-4-fluorobenzamide (Cpd 48); 3-((4-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)-4-fluorobenzamide

5-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-2,6-dione (Cpd 49);

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 50); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)isoindole-1,3-dione (Cpd 51); 5-(4-((1-((1-((2,3-dihydrobenzo[b][1,4]dioxadiene-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-5-yl

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 52); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(pyridin-2-ylmethyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)isoindole-1,3-dione (Cpd 53); 3-((4-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 54); 3-((4-(4-((1-((1-butyl-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-2,6-dione (Cpd 55);

5-(4-((1-((1-((1,3-dioxolan-2-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-2,6-dione (Cpd 55);

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 56); 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)isoindole-1,3-dione (Cpd 57); 3-((4-(4-((1-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-1,3-dione

-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 58); 5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 59); 3-((4-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 60); 4-amino-N-(3-(5-((4-((4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-1-yl)propyl)piperidine-4-carboxamide (Cpd 61); 5-((4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-inden-5-yl)piperidin-

-1-yl)methyl)piperidin-1-yl)methyl)-N,N-dimethyl-3-(4-(trifluoromethoxy)phenyl)-1H-indole-1-sulfonamide (Cpd 62); 3-((3-fluoro-4-(4-((1-((1-(4-methylpentyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-5-yl)methyl)piperidin-1-yl

-1-yl)phenyl)amino)piperidin-2,6-dione (Cpd 63); 3-((4-fluoro-3-(4-((1-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-2,6-dione

-1-yl)phenyl)amino)piperidine-2,6-dione (Cpd 64); 3-((3-fluoro-4-(1'-((3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidinyl]-1-yl)phenyl)amino)piperidine-2,6-dione (Cpd 65); 5-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 66); 3-((4-(5-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (Cpd 67); 5-(4-((1-((1-(3,3-dimethoxypropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-2,6-dione

-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 68); 5-(5-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 69); 5-(1'-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[3,3'-azocyclobutane]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 70); 5-(1'-((1-(3-(dimethylamino)propyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)-[4,4'-bipiperidin]-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (Cpd 71); (2R,4S)-1-((R)-2-(2-(4-((1-((1-(3-aminopropyl)-3-(4-(trifluoromethoxy)phenyl)-1H-indol-5-yl)methyl)piperidin-4-yl)methyl)piperidin-

-1-yl)acetamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((R)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Cpd 72); or a pharmaceutically acceptable salt, hydrate, tautomer or stereoisomer thereof. A pharmaceutical composition comprising an effective amount of a bifunctional compound as claimed in any one of claims 1 to 15 and one or more pharmaceutically acceptable excipients. A pharmaceutical composition for preventing, ameliorating and/or treating a disease associated with KRAS mutation in an individual in need thereof, comprising an effective amount of a bifunctional compound as described in any one of claims 1 to 15 and one or more pharmaceutically acceptable excipients. A pharmaceutical composition as claimed in claim 17, wherein the bifunctional compound is administered separately, sequentially or together with one or more other anticancer agents. The pharmaceutical composition of claim 18, wherein the other anticancer agents are selected from the group consisting of: trastuzumab, ramucirumab, vismodegib, sonidegib, bevacizumab, everolimus, tamoxifen, toremifene, fulvestrant, anastrozole, exemestane, lapatinib, letrozole, pertuzumab, ado-trastuzumab emtansine, palbociclib, cetuximab, panitumumab, ziv-aflibercept, regorafenib, lmatinib mesylate, lanreotide acetate, sunitinib, denosumab, alitretinoin, sorafenib, pazopanib, temsirolimus, everolimus, tretinoin, dasatinib, nilotinib, bosutinib, rituximab uximab), alemtuzumab, ofatumumab, obinutuxumab, ibrutinib, idelalisib, blinatumomab, soragenib, crizotinib, erlotinib, gefitinib, afatinib dimaleate, ceritnib, ramucirumab, nivolumab, pembrolizumab, osimertinib, necitumumab, busulfan, chlorambucil, cyclophosphamide, iphosphamide, melphalan, nitrogen mustard, streptozocin, thiotepa, uracil nitrogen mustard mustard), triethylenemelamine, temozolomide, SarCNU, actinomycin-D, bleomycin, cryptophycins, daunorubicin, doxorubicin, idarubicin, irinotecan, L-asparaginase, mitomycin-C, mitramycin, navelbine, paclitaxel, litaxel, docetaxel, topotecan, vinblastine, vincristine, teniposide (VM-26), etoposide (VP-16), 5α-reductase inhibitors, aminoglutethimide, anastrozole, bicalutamide, chlorotrianisene, diethylstilbestrol (DES), dromostanolone, estramustine, ethinyl estradiol, estradiol), flutamide, fluoxymesterone, goserelin, hydroxyprogesterone, letrozole, leuprolide, medroxyprogesterone acetate, megestrol acetate, methylprogesterone acetate, prednisolone, methyltestosterone, mitotane, nilutamide, prednisolone, arzoxifene (SERM-3), tamoxifen, testolactone, testosterone, triamicnolone, zoladex, all-trans retinoic acid, carmustine (BCNU), carboplatin n)(CBDCA), lomustine (CCNU), cis-diaminedichloroplatinum (cis-platinum), dacarbazine, gliadel, hexamethylmelamine, hydroxyurea, levamisole, mitoxantrone, o,p'-dichlorodiphenyldichloroethane (o,p'-DDD) (also known as lysodren or mitotane), oxaliplatin, porfimer sodium sodium), procarbazine, imatinib mesylate, chlorodeoxyadenosine, cytosine arabinoside, 2'-deoxycoformycin, fludarabine phosphate, 5-fluorouracil (5-FU), 5-fluoro-2'-deoxyuridine (5-FUdR), gemcitabine, camptothecin, 6-mercaptopurine, methotrexate, 4-methylthioamphetamine (4-MTA), thioguanine, alpha-interferon, Bacillus Calmette-Guérin (BCG) Calmette-Guerin; BCG), granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-2 and herceptin. The pharmaceutical composition of claim 17, wherein the disease is cancer, autoimmune disease, infectious disease or angiogenic disease. The pharmaceutical composition of claim 20, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, bladder cancer, stomach cancer, kidney cancer, salivary gland cancer, ovarian cancer, uterine cancer, cervical cancer, oral cancer, skin cancer, brain cancer, lymphoma, leukemia, bile duct malignancy and endometrial cancer. The pharmaceutical composition of claim 21, wherein the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer or myeloid leukemia.