WO2025250897A1 - Heterobifunctional compounds and methods of use thereof - Google Patents

Abstract

Disclosed herein are heterobifunctional compounds that include a moiety that binds to the Y220C mutant of the p53 protein, a moiety that binds to the effector protein BRD4, and a linker. Also disclosed herein are pharmaceutical compositions comprising the compounds, and methods of using the compounds, e.g., to treat proliferative diseases such as cancers.

Description

HETEROBIFUNCTIONAL COMPOUNDS AND METHODS OF USE THEREOF

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63/654,403, filed on May 31, 2024, the disclosure of which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

Disclosed herein are heterobifunctional compounds that include a moiety that binds to the Y220C mutant of the p53 protein, a moiety that binds to the effector protein BRD4, and a linker. Also disclosed herein are pharmaceutical compositions comprising the compounds, and methods of using the compounds, e.g., to treat proliferative diseases such as cancers.

BACKGROUND p53 is a nuclear transcription factor that has tumor suppression function. Its function is commonly disrupted in cancer cells via p53 gene mutation that leads to either loss of DNA binding or low protein thermostability. Genome sequencing of different cancers showed that 42% of cases carried a mutation in p53 (Wang et al. Signal Transcluct. Target Ther. 8(1):92 (2023)). The Y220C mutation, representing 1.3% of all p53 mutations, results in formation of a hydrophobic cavity on the p53 surface. Small molecules can bind to this hydrophobic cavity and restore p53 function. This occupancy-based approach, however, results in poor target engagement and high cytotoxicity.

SUMMARY

In one aspect, disclosed herein is a compound of formula (I) BB-L-PB (I) or a pharmaceutically acceptable salt thereof, wherein: BB is moiety that binds to BRD4;

L is a linker; and

PB is a moiety that binds to a Y220C mutant of p53.

In some embodiments, BB has a formula selected from:

In some embodiments, BB is a moiety of formula:

In some embodiments, PB is a moiety of formula:

In some embodiments, PB is a moiety of formula:

5

In some embodiments, the compound is a compound of formula (la): or a pharmaceutically acceptable salt thereof.

In some embodiments, L is a direct bond or comprises any combination of -CH2-, -

CH=CH-, -C=C-, -O-, -NR'-, -BR'-, -S-, -C(O)-, -C(NR')-, -S(O)-, -S(O)₂-, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents. In some embodiments, L is a direct bond or comprises any combination of -CH2-, -O-, -C(O)-, -NH-, -N(CH3)-, cycloalkylene, and heterocyclylene moieties. In some embodiments, L comprises any combination of the following moieties; wherein p is 1, 2, 3, 4, 5, or 6; and q is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

In some embodiments, L has a formula selected from:

In some embodiments, the compound is selected from:

and pharmaceutically acceptable salts thereof. In another aspect, disclosed herein is a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of formula (I) or a compound of formula (la), or a pharmaceutically acceptable salt thereof), and a pharmaceutically acceptable carrier.

In another aspect, disclosed herein is a method of treating a proliferative disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula (I) or a compound of formula (la), or a pharmaceutically acceptable salt thereof).

In some embodiments, the proliferative disease is a cancer selected from a carcinoma, a sarcoma, and a hematologic malignancy. In some embodiments, the cancer is selected from thyroid cancer, lung cancer, breast cancer, liver cancer, stomach cancer, pancreatic cancer, colorectal cancer, kidney cancer, urethral cancer, ovarian cancer, prostate cancer, bone cancer, bladder cancer, esophageal cancer, brain cancer, head and neck cancer, skin cancer, a lymphoma, a leukemia, and multiple myeloma. In one aspect, disclosed herein is a compound disclosed herein (e.g., a compound of formula (I) or a compound of formula (la), or a pharmaceutically acceptable salt thereof), for use as a medicament.

In another aspect, disclosed herein is a compound disclosed herein (e.g., a compound of formula (I) or a compound of formula (la), or a pharmaceutically acceptable salt thereof), for use in treating a proliferative disease. In some embodiments, the proliferative disease is a cancer selected from a carcinoma, a sarcoma, and a hematologic malignancy. In some embodiments, the cancer is selected from thyroid cancer, lung cancer, breast cancer, liver cancer, stomach cancer, pancreatic cancer, colorectal cancer, kidney cancer, urethral cancer, ovarian cancer, prostate cancer, bone cancer, bladder cancer, esophageal cancer, brain cancer, head and neck cancer, skin cancer, a lymphoma, a leukemia, and multiple myeloma.

In another aspect, disclosed herein is a kit comprising a compound disclosed herein (e.g., a compound of formula (I) or a compound of formula (la), or a pharmaceutically acceptable salt thereof).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a scheme illustrating a bivalent compound containing a moiety that binds to the p53 Y220C mutant and a moiety that binds to BRD4, which can reactivate p53 transcription.

FIG. 2 shows the activation of the p53 Y220C reporter following a 24-hour treatment with bivalent compounds described herein.

FIG. 3 demonstrates the anti-proliferative effects of bivalent compounds disclosed herein on BxPC-3, a pancreatic cancer cell line.

FIG. 4 shows the formation of a ternary complex between p53 Y220C and BRD4, induced by bivalent compounds.

FIG. 5 shows the expression of representative p53 target genes after a 16-hour treatment with bivalent compounds.

FIG. 6 shows the expression of the p21 following a 2-hour treatment with bivalent compounds.

FIG. 7 shows the proliferation rate of cells treated with bivalent compounds for a short period of time. FIG. 8 shows the difference in p53 Y220C reporter activation between negative control compounds, which can bind only to one protein, and bivalent compounds.

DETAILED DESCRIPTION

Disclosed herein are bivalent compounds that target p53 transcription activity via an event-driven mechanism. The compounds include moiety that binds to the BRD4 protein and a moiety that binds to the Y220C mutant of the p53 protein, which are connected via a linker. These bivalent compounds can induce proximity between BRD4 and the Y220C p53 protein, such that the p53 binder will reactivate p53 transcription and BRD4 will enhance the transcription activity through the proximity-based interaction. See, e.g., FIG. 1 . By modulating p53 target gene transcription activity, the compounds can induce strong transcription activity and potent anti-proliferation activity in cancer cell lines. Pharmaceutical compositions comprising the disclosed compounds, methods of using the disclosed compounds, and kits comprising the compounds are also disclosed herein.

Definitions

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control.

Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Sorrell, Organic Chemistry, 2^nd edition, University Science Books, Sausalito, 2006; Smith, March’s Advanced Organic Chemistry: Reactions, Mechanism, and Structure, 7^th Edition, John Wiley & Sons, Inc., New York, 2013; Larock, Comprehensive Organic Transformations, 3^rd Edition, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3^rd Edition, Cambridge University Press, Cambridge, 1987.

As used herein, the modifier “about” used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context (for example, it includes at least the degree of error associated with the measurement of the particular quantity). The modifier “about” should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the expression “from about 2 to about 4” also discloses the range “from 2 to 4.” The term “about” may refer to ±10% of the indicated number. For example, “about 10%” may indicate a range of 9% to 11%, and “about 1” may mean from 0.9 - 1.1. Other meanings of “about” may be apparent from the context, such as rounding off; for example, “about 1” may also mean from 0.5 to 1.4.

As used herein, the term “alkyl” refers to a radical of a straight or branched saturated hydrocarbon chain. The alkyl chain can include, e.g., from 1 to 24 carbon atoms (C1-C24 alkyl), 1 to 16 carbon atoms (C1-C16 alkyl), 1 to 14 carbon atoms (C1-C14 alkyl), 1 to 12 carbon atoms (Ci- C12 alkyl), 1 to 10 carbon atoms (C1-C10 alkyl), 1 to 8 carbon atoms (Ci-Cs alkyl), 1 to 6 carbon atoms (Ci-Ce alkyl), 1 to 4 carbon atoms (C1-C4 alkyl), 1 to 3 carbon atoms (C1-C3 alkyl), or 1 to 2 carbon atoms (C1-C2 alkyl). Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n- nonyl, n-decyl, n-undecyl, and n-dodecyl.

As used herein, the term “alkenyl” refers to a radical of a straight or branched hydrocarbon chain containing at least one carbon-carbon double bond and no triple bonds. The double bond(s) may be located at any position(s) with the hydrocarbon chain. The alkenyl chain can include, e.g., from 2 to 24 carbon atoms (C2-C24 alkenyl), 2 to 16 carbon atoms (C2-C16 alkenyl), 2 to 14 carbon atoms (C2-C14 alkenyl), 2 to 12 carbon atoms (C2-C12 alkenyl), 2 to 10 carbon atoms (C2-C10 alkenyl), 2 to 8 carbon atoms (C2-C8 alkenyl), 2 to 6 carbon atoms (C2-C6 alkenyl), 2 to 4 carbon atoms (C2-C4 alkenyl), 2 to 3 carbon atoms (C2-C3 alkenyl), or 2 carbon atoms (C2 alkenyl). Representative examples of alkenyl include, but are not limited to, ethenyl, 1 -propenyl, 2-propenyl, 1-butenyl, 2-butenyl, butadienyl, 2-methyl-2-propenyl, 3-butenyl, pentenyl, pentadienyl, hexenyl, heptenyl, octenyl, octatrienyl, and the like.

As used herein, the term “alkynyl” means a radical of a straight or branched hydrocarbon chain containing at least one carbon-carbon triple bond. The alkynyl chain can include, e.g., from 2 to 24 carbon atoms (C2-C24 alkynyl), 2 to 16 carbon atoms (C2-C16 alkynyl), 2 to 14 carbon atoms (C2-C14 alkynyl), 2 to 12 carbon atoms (C2-C12 alkynyl), 2 to 10 carbon atoms (C2-C10 alkynyl), 2 to 8 carbon atoms (C2-C8 alkynyl), 2 to 6 carbon atoms (C2-C6 alkynyl), 2 to 4 carbon atoms (C2-C4 alkynyl), 2 to 3 carbon atoms (C2-C3 alkynyl), or 2 carbon atoms (C2 alkynyl). The triple bond(s) may be located at any position(s) with the hydrocarbon chain. Representative examples of alkynyl include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and the like.

As used herein, the term “alkoxy” refers to an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy.

As used herein, the term “amino” refers to a group -NR_xR_y, wherein R_x and R_y are selected from hydrogen and alkyl (e.g., C1-C4 alkyl). A group -NH(alkyl) may be referred to herein as “alkylamino” and a group -N(alkyl)2 may be referred to herein as “dialkylamino.”

As used herein, the term “aryl” refers to a radical of a monocyclic, bicyclic, or tricyclic 4n+2 aromatic ring system (e.g., having 6, 10, or 14 it electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms (“C6-C14 aryl”). In some embodiments, an aryl group has six ring carbon atoms (“Ce aryl,” i.e., phenyl). In some embodiments, an aryl group has ten ring carbon atoms (“C10 aryl,” e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms (“C14 aryl,” e.g., anthracenyl and phenanthrenyl).

As used herein, the term “arylene” refers to a divalent aryl radical.

As used herein, the term “cycloalkyl” refers to a radical of a saturated carbocyclic ring system containing three to ten carbon atoms and zero heteroatoms. The cycloalkyl may be monocyclic, bicyclic, bridged, fused, or spirocyclic. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl, bicyclo[2.2.1]heptanyl, bicyclo[3.2.1]octanyl, and bicyclo[5.2.0]nonanyl.

As used herein, the term “cycloalkylene” refers to a divalent cycloalkyl radical.

As used herein, the term “cyano” refers to a -CN group.

As used herein, the term “halogen” or “halo” refers to F, Cl, Br, or I.

As used herein, the term “haloalkyl” refers to an alkyl group, as defined herein, in which at least one hydrogen atom (e.g., one, two, three, four, five, six, seven or eight hydrogen atoms) is replaced with a halogen. In some embodiments, each hydrogen atom of the alkyl group is replaced with a halogen (“perhaloalkyl”). Representative examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, and 3,3,3-trifluoropropyl.

As used herein, the term “haloalkoxy” refers to a haloalkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom. Representative examples of haloalkoxy include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2,2,2- trifluoroethoxy.

As used herein, the term “heteroaryl” refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 n electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-10 membered heteroaryl”). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused (aryl/heteroaryl) ring system. Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, and the like) the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl). Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5- membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6-membered hctcroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7- membered hctcroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotri azolyl, benzothiophenyl, isobenzothiophenyl, bcnzofuranyl, bcnzoisofuranyl, bcnzimidazolyl, bcnzoxazolyl, bcnzisoxazolyl, bcnzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6- bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

As used herein, the term “hetero arylene” refers to a divalent heteroaryl radical.

As used herein, the term “heterocyclyl” refers to a radical of a 3- to 10-membered nonaromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and can be saturated or can be partially unsaturated. Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more cycloalkyl groups wherein the point of attachment is either on the cycloalkyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. A heterocyclyl group may be described as, e.g., a 3-7-membered heterocyclyl, wherein the term “membered” refers to the non-hydrogen ring atoms, i.e., carbon, nitrogen, oxygen, sulfur, boron, phosphorus, and silicon, within the moiety. Exemplary 3-membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, and thiorenyl. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclyl groups containing one heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2, 5-dione. Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5- membered heterocyclyl groups containing three heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing one hctcroatom include, without limitation, pipcridinyl (c.g., 2, 2,6,6- tetramethylpiperidinyl), tetrahydropyranyl, dihydropyridinyl, pyridinonyl (e.g., 1-methylpyridin- 2-onyl), and thianyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, pyridazinonyl (2-methylpyridazin-3-onyl), pyrimidinonyl (e.g., l-methylpyrimidin-2-onyl, 3-methylpyrimidin-4-onyl), dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary 5-membered heterocyclyl groups fused to a Ce aryl ring (also referred to herein as a 5,6-bicyclic heterocyclyl ring) include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 5- membered heterocyclyl groups fused to a heterocyclyl ring (also referred to herein as a 5,5- bicyclic heterocyclyl ring) include, without limitation, octahydropyrrolopyrrolyl (e.g., octahydropyrrolo[3,4-c]pyrrolyl), and the like. Exemplary 6-membered heterocyclyl groups fused to a heterocyclyl ring (also referred to as a 4,6-membered heterocyclyl ring) include, without limitation, diazaspirononanyl (e.g., 2,7-diazaspiro[3.5]nonanyl). Exemplary 6-membered heterocyclyl groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocyclyl ring) include, without limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. Exemplary 6-membered heterocyclyl groups fused to a cycloalkyl ring (also referred to herein as a 6,7-bicyclic heterocyclyl ring) include, without limitation, azabicyclooctanyl (e.g., (1 ,5)-8- azabicyclo[3.2.1]octanyl). Exemplary 6-membered heterocyclyl groups fused to a cycloalkyl ring (also referred to herein as a 6,8-bicyclic heterocyclyl ring) include, without limitation, azabicyclononanyl (e.g., 9-azabicyclo[3.3.1]nonanyl).

As used herein, the term “heterocyclylene” refers to a divalent heterocyclyl radical. As used herein, the term “hydroxy” or “hydroxyl” refers to an -OH group. As used herein, the term “nitro” refers to an -NO2 group.

When a group or moiety can be substituted, the term “substituted” indicates that one or more (e.g., 1, 2, 3, 4, 5, or 6; in some embodiments 1, 2, or 3; and in other embodiments 1 or 2) hydrogens on the group indicated in the expression using “substituted” can be replaced with a selection of recited indicated groups or with a suitable substituent group known to those of skill in the art (c.g., one or more of the groups recited below), provided that the designated atom’s normal valence is not exceeded. Substituent groups include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amido, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, cycloalkyl, cycloalkenyl, guanidino, halo, haloalkyl, haloalkoxy, heteroaryl, heterocyclyl, hydroxy, hydrazino, imino, oxo, nitro, phosphate, phosphonate, sulfonic acid, thiol, thione, or combinations thereof.

As used herein, in chemical structures the indication: represents a point of attachment of one moiety to another moiety (e.g., a substituent group to the rest of the compound).

For compounds described herein, groups and substituents thereof may be selected in accordance with permitted valence of the atoms and the substituents, such that the selections and substitutions result in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

When substituent groups are specified by their conventional chemical formulae, written from left to right, such indication also encompass substituent groups resulting from writing the structure from right to left. For example, if a bivalent group is shown as -CH2O-, such indication also encompasses -OCH2-; similarly, -OC(O)NH- also encompasses -NHC(O)O-. When linker moieties are shown, the linkers can be attached to other moieties of the compound in either direction.

The terms “administer,” “administering,” or “administration,” as used herein refer to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound or a pharmaceutical composition.

As used herein, the terms “condition,” “disease,” and “disorder” are used interchangeably .

An “effective amount” of a compound or composition refers to an amount sufficient to elicit a desired biological response (e.g., treating a condition). As will be appreciated by those skilled in the ail, the effective amount of a compound may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. An effective amount encompasses therapeutic and prophylactic treatment. For example, in treating cancer, an effective amount of a compound or composition may reduce tumor burden or stop the growth or spread of a tumor.

A “therapeutically effective amount” of a compound or composition is an amount sufficient to provide a therapeutic benefit in the treatment of a condition, or to delay or minimize one or more symptoms associated with the condition. In some embodiments, a therapeutically effective amount is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to minimize one or more symptoms associated with the condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of the condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of the condition, or enhances the therapeutic efficacy of another therapeutic agent.

A “subject” to which administration is contemplated includes, but is not limited to, a human (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) and/or other non-human animals, for example, mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs) and birds (e.g., commercially relevant birds such as chickens, ducks, geese, and/or turkeys).

As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or condition, or one or more signs or symptoms thereof. In some embodiments, “treatment,” “treat,” and “treating” require that signs or symptoms of the disease disorder or condition have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease or condition. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.

Compounds

Disclosed herein is a compound of formula (I): BB-L-PB (I) or a pharmaceutically acceptable salt thereof, wherein: BB is moiety that binds to BRD4;

L is a linker; and PB is a moiety that binds to a Y220C mutant of p53.

In embodiments, BB has a formula selected from: In some embodiments, BB is a BRD4-binding moiety of formula:

In some embodiments, PB is a moiety of formula:

In some embodiments, PB is a moiety of formula:

In some embodiments, PB is a moiety of formula:

In some embodiments, the compound is a compound of formula (la):

or a pharmaceutically acceptable salt thereof.

L is a linker, which provides a covalent attachment between the moiety that binds to BRD4 and the moiety that binds to the Y220C p53. The exact structure of linker may not be critical, provided it is substantially non-interfering with the activities of the other moieties. In some embodiments, L is a direct bond. In other embodiments, L comprises any combination of - CH2-, -CH=CH-, -C=C-, -O-, -NR'-, -BR'-, -S-, -C(O)-, -C(NR')-, -S(O)-, -S(O)₂-, arylene, hctcroarylcnc, cycloalky lenc, and hctcrocyclylcnc moieties, wherein R' is selected from hydrogen and Ci-Ce alkyl, and wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents.

In some embodiments, the linker comprises an alkylene chain (e.g., having 2-20 -CH2- units). In other embodiments, the linker comprises an alkylene chain that is interrupted by, and/or terminated by (at either or both termini), at least one group selected from -O-, -S-, -N(R')- , -CH=CH-, -C=C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, - C(O)N(R’)C(O)-, -C(O)N(R')C(O)N(R')-, -N(R’)C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, - OC(O)N(R')-, -C(NR’)-, -N(R')C(NR')-, -C(NR')N(R’)-, -N(R')C(NR')N(R’)-, -OB(CH₃)O-, - S(O)₂-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)₂-, -S(O)₂O-, -N(R')S(O)₂-, -S(O)₂N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)₂N(R')-, -N(R’)S(O)N(R')-, C3-C12 cycloalkylene, 3- to 12-membered heterocyclylene, 5- to 10-membered arylene, 5- to 12-membered heteroarylene, or any combination thereof, wherein each R' is independently selected from hydrogen and Ci-Ce alkyl, and wherein the interrupting and terminating groups may be the same or different.

In some embodiments, L comprises an alkylene chain: wherein q is 1, 2, 3, 4,

5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. In some embodiments, q is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. For example in some embodiments, q is 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 1-9, 1-8, 1 -7, 1-6, 1 -5, 1-4, 1-3, 1-2, 2-16, 2-15, 2- 14, 2-13, 2-12, 2-11 , 2-10, 2-9, 2-8, 2-7, 2-6, 2-5,

2-4, 2-3, 3-16, 3-15, 3-14, 3-13, 3-12, 3-11, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-16, 4-15, 4-14, 4- 13, 4-12, 4-11, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-16, 5-15, 5-14, 5-13, 5-12, 5-11, 5-10, 5-9, 5-8, 5- 7, 5-6, 6-16, 6-15, 6-14, 6-13, 6-12, 6-11, 6-10, 6-9, 6-8, 6-7, 7-16, 7-15, 7-14, 7-13, 7-12, 7-11, 7-10, 7-9, 7-8, 8-16, 8-15, 8-14, 8-13, 8-12, 8-11, 8-10, 8-9, or 9-16, 9-15, 9-14, 9-13, 9-12, 9-11, 9-10, 10-16, 10-15, 10-14, 10-13, 10-12, 10-11, 11-16, 11-15, 11-14, 11-13, 11-12, 12-16, 12-15, 12-14, 12-13, 13-16, 13-15, 13-14, 14-16, 14-15, and 15-16. In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3. In some embodiments, q is 4. In some embodiments, q is 5. In some embodiments, q is 6. In some embodiments, q is 7. In some embodiments, q is 8. In some embodiments, q is 9. In some embodiments, q is 10. In some embodiments, q is 11. In some embodiments, q is 12. In some embodiments, q is 13. In some embodiments, q is 14. In some embodiments, q is 15. In some embodiments, q is 16. Specific examples of alkylene chains, include:

In some embodiments, L comprises an alkylene chain interrupted by a functional group, such as -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, - C(O)N(R')C(O)-, - C(O)N(R')C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R’)C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR’)N(R’)-, -OB(CH₃)O-, -S(O)₂-, -OS(O)-, -S(O)O-, - S(O)-, -OS(O)₂-, -S(O)₂O-, -N(R’)S(O)2-, -S(O)₂N(R')-, -N(R')S(O)-, -S(O)N(R')-, - N(R')S(O)₂N(R')-, or -N(R')S(O)N(R')-.

In some embodiments, L comprises one or more alkylene glycol repeat units, such as ethylene glycol or propylene glycol repeat units. For example, in some embodiments, L comprises a poly- or oligo-ethylene glycol chain: , wherein p is 1, 2, 3, 4, 5, or 6. In some embodiments, L comprises a group of formula: , wherein p is 1, 2, 3, 4,

5, or 6. In some embodiments, p is 1-6, 1-5, 1-4, 1-3, 1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-

6, or 5-6. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6.

In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups, non-limiting examples of which include:

In some embodiments, L comprises a cycloalkylene or heterocyclylene group, such as one of the above-illustrated cycloalky leno and heterocyclylene groups, with one or more additional groups on one or both termini, wherein the additional groups are independently selected from -CH₂-, -O-, -S-, -N(R')-, -CH=CH-, -C=C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, - C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R’)C(O)-, - N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR’)N(R’)-, - N(R')C(NR')N(R')-, -OB(CH₃)O-, -S(0)₂-, -0S(0)-, -S(0)0-, -S(0)-, -0S(0)₂-, -S(0)₂0-, - N(R')S(O)₂-, -S(O)₂N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)₂N(R')-, and -N(R')S(O)N(R')-, or any combination thereof, wherein each R' is independently selected from hydrogen and Ci-Ce alkyl. In some embodiments, L comprises a cycloalkylene or heterocyclylene group, such as one of the above-illustrated cycloalkylene and heterocyclylene groups, with one or more additional groups on one or both termini, wherein the additional groups are independently selected from - CH₂-, -C(O)-, -NH-, and -O-.

In some embodiments, L comprises two or more cycloalkylene or heterocyclylene groups, such as two or three cycloalkylene or heterocyclylene groups independently selected from those illustrated above, connected to each other via covalent bonds or one or linking groups, optionally with one or more additional linking group son one or both termini, wherein the linking groups are independently selected from -O-, -S-, -N(R')-, -CH₂-, -CH=CH-, -C=C-, - C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, - C(O)N(R')C(O)-, - C(O)N(R')C(O)N(R')-, -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R’)-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R’)-, -OB(CH₃)O-, -S(O)₂-, -OS(O)-, -S(O)O-, - S(O)-, -OS(O)2-, -S(O)₂O-, -N(R’)S(O)₂-, -S(O)₂N(R’)-, -N(R')S(O)-, -S(O)N(R')-, - N(R')S(O)₂N(R')-, and -N(R')S(O)N(R')-, or any combination thereof, wherein each R' is independently selected from hydrogen and Ci-Ce alkyl. In some embodiments, L comprises two or more cycloalkylene or heterocyclylene groups, such as two or three cycloalkylene or heterocyclylene groups independently selected from those illustrated above, connected to each other via covalent bonds or one or linking groups, optionally with one or more additional linking group son one or both termini, wherein the linking groups are independently selected from -O- and -CH2-.

In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups that are substituted with 1, 2, or 3 substituents, e.g., substituents independently selected from Ci- C4 alkyl, C1-C4 haloalky 1, C1-C4 alkoxy, halo, hydroxy, and the like. In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups that are substituted with 1, 2, or 3 substituents independently selected from C1-C4 alkyl and halo. In some embodiments, L comprises one or more cycloalkylene or heterocyclylene groups that are substituted with 1, 2, or 3 substituents independently selected from methyl and fluoro.

In some embodiments, L comprises any combination of the following moieties:

In some embodiments, the compound is selected from:

and pharmaceutically acceptable salts thereof.

When discussing certain features or properties of compounds of formula (I) herein, or compositions, methods, or kits comprising compounds of formula (I), it is understood that such reference also includes compounds of formula (la), and to specific exemplary compounds disclosed herein.

The compound may exist as a stereoisomer wherein asymmetric or chiral centers are present. The stereoisomer is “R” or “S” depending on the configuration of substituents around the chiral carbon atom. The terms “R” and “S” used herein are configurations as defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem. 1976, 45: 13-30. Various stereoisomers and mixtures thereof are specifically included within the scope of this disclosure. Stereoisomers include enantiomers and diastereomers, and mixtures of enantiomers or diastereomers. Individual stereoisomers of the compounds may be prepared synthetically from commercially available stalling materials, which contain asymmetric or chiral centers or by preparation of racemic mixtures followed by methods of resolution well- known to those of ordinary skill in the art. These methods of resolution are exemplified by: (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and optional liberation of the optically pure product from the auxiliary as described in Furniss, Hannaford, Smith, and Tatchell, “Vogel's Textbook of Practical Organic Chemistry,” 5th edition (1989), Longman Scientific & Technical, Essex CM202JE, England; (2) direct separation of the mixture of optical enantiomers on chiral chromatographic columns; or (3) fractional recrystallization methods.

It should be understood that the compounds may exist in different tautomeric forms, and all such forms are included within the scope of the disclosure.

The present disclosure also includes an isotopically-labeled compound, which is identical to those recited in formula (I), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention arc hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as, but not limited to ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷0, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶C1, respectively. Substitution with heavier isotopes such as deuterium (²H) can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. The compound may incorporate positron-emitting isotopes for medical imaging and positron-emitting tomography (PET) studies for determining the distribution of receptors. Suitable positron-emitting isotopes that can be incorporated in compounds of formula (I) are ⁿC, ¹³N, ¹⁵O, and ¹⁸F. Isotopically- labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples using appropriate isotopically-labeled reagent in place of non-isotopically-labeled reagent.

Compounds disclosed herein can exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the disclosure encompass both solvated and unsolvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in a crystalline form. a. Pharmaceutically Acceptable Salts

The disclosed compounds may exist as pharmaceutically acceptable salts. The term “pharmaceutically acceptable salt” refers to salts or zwitterions of the compounds which are water or oil-soluble or dispersible, suitable for treatment of disorders without undue toxicity, irritation, and allergic response, commensurate with a reasonable benefit/risk ratio and effective for their intended use. The salts may be prepared during the final isolation and purification of the compounds or separately by reacting an amino group of the compounds with a suitable acid. For example, a compound may be dissolved in a suitable solvent, such as but not limited to methanol and water and treated with at least one equivalent of an acid, like hydrochloric acid. The resulting salt may precipitate out and be isolated by filtration and dried under reduced pressure. Alternatively, the solvent and excess acid may be removed under reduced pressure to provide a salt. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hcmisulfatc, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3 -phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, para-toluenesulfonate, undecanoate, hydrochloric, hydrobromic, sulfuric, phosphoric, and the like. Amino groups of the compounds may also be quatemized with alkyl chlorides, bromides and iodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl and the like.

Basic addition salts may be prepared during the final isolation and purification of the disclosed compounds by reaction of a carboxyl group with a suitable base such as the hydroxide, carbonate, or bicarbonate of a metal cation such as lithium, sodium, potassium, calcium, magnesium, or aluminum, or an organic primary, secondary, or tertiary amine. Quaternary amine salts can be prepared, such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N- methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N- dibenzylphenethylamine, 1 -ephenamine and N,N’ -dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like. b. Methods of Synthesis

In another aspect, disclosed herein are methods for making compounds of formula (I), or a pharmaceutically acceptable salt thereof. Broadly, the compounds of formula (I) and pharmaceutically acceptable salts thereof can be prepared by any process known to be applicable to the preparation of chemically related compounds. Exemplary suitable synthetic schemes are provided in the Examples section.

The compounds and intermediates may be isolated and purified by methods well-known to those skilled in the art of organic synthesis. Examples of conventional methods for isolating and purifying compounds can include, but are not limited to, chromatography on solid supports such as silica gel, alumina, or silica derivatized with alkylsilane groups, by recrystallization at high or low temperature with an optional pretreatment with activated carbon, thin-layer chromatography, distillation at various pressures, sublimation under vacuum, and trituration, as described for example in “Vogel’s Textbook of Practical Organic Chemistry,” 5th edition (1989), by Fumiss, Hannaford, Smith, and Tatchell, pub. Longman Scientific & Technical, Essex CM20 2JE, England.

Reaction conditions and reaction times for each individual step can vary depending on the particular reactants employed and substituents present in the reactants used. Reactions can be worked up in a conventional manner, e.g., by eliminating the solvent from the residue and further purified according to methodologies generally known in the art such as, but not limited to, crystallization, distillation, extraction, trituration and chromatography. Unless otherwise described, the starting materials and reagents are either commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature.

Routine experimentations, including appropriate manipulation of the reaction conditions, reagents and sequence of the synthetic route, protection of any chemical functionality that cannot be compatible with the reaction conditions, and deprotection at a suitable point in the reaction sequence of the method, are included in the scope of the disclosure. Suitable protecting groups and the methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which can be found in PGM Wuts and TW Greene, in Greene’s book titled Protective Groups in Organic Synthesis (4^th ed.), John Wiley & Sons, NY (2006).

When an optically active form of a disclosed compound is required, it can be obtained by carrying out one of the procedures described herein using an optically active starting material (prepared, for example, by asymmetric induction of a suitable reaction step), or by resolution of a mixture of the stereoisomers of the compound or intermediates using a standard procedure (such as chromatographic separation, recrystallization or enzymatic resolution).

Similarly, when a pure geometric isomer of a compound is required, it can be obtained by carrying out one of the procedures described herein using a pure geometric isomer as a starting material, or by resolution of a mixture of the geometric isomers of the compound or intermediates using a standard procedure such as chromatographic separation.

The synthetic schemes and specific examples as described are illustrative and are not to be read as limiting the scope of the disclosure or the claims. Alternatives, modifications, and equivalents of the synthetic methods and specific examples are contemplated. Pharmaceutical Compositions

The disclosed compounds may be incorporated into pharmaceutical compositions suitable for administration to a subject (such as a patient, which may be a human or non-human). The pharmaceutical compositions may include a “therapeutically effective amount” or a “prophylactic ally effective amount” of the agent. A “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the composition may be determined by a person skilled in the art and may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of a compound of the disclosure are outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease or condition, the prophylactically effective amount will be less than the therapeutically effective amount.

The pharmaceutical compositions may include pharmaceutically acceptable carriers. The term “pharmaceutically acceptable earner,” as used herein, means a non-toxic, inert solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as, but not limited to, lactose, glucose and sucrose; starches such as, but not limited to, com starch and potato starch; cellulose and its derivatives such as, but not limited to, sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as, but not limited to, cocoa butter and suppository waxes; oils such as, but not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols; such as propylene glycol; esters such as, but not limited to, ethyl oleate and ethyl laurate; agar; buffering agents such as, but not limited to, magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as, but not limited to, sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. Thus, the compounds and their pharmaceutically acceptable salts may be formulated for administration by, for example, solid dosing, eye drop, in a topical oil-based formulation, injection, inhalation (either through the mouth or the nose), implants, or oral, buccal, parenteral, or rectal administration. Techniques and formulations may generally be found in “Remington’s Pharmaceutical Sciences,” (Meade Publishing Co., Easton, Pa.). Therapeutic compositions must typically be sterile and stable under the conditions of manufacture and storage.

The route by which the disclosed compounds are administered and the form of the composition will dictate the type of carrier to be used. The composition may be in a variety of forms, suitable, for example, for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implants, or parenteral) or topical administration (e.g., dermal, pulmonary, nasal, aural, ocular, liposome delivery systems, or iontophoresis).

Carriers for systemic administration typically include at least one of diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, combinations thereof, and others. All carriers are optional in the compositions.

Suitable diluents include sugars such as glucose, lactose, dextrose, and sucrose; diols such as propylene glycol; calcium carbonate; sodium carbonate; sugar alcohols, such as glycerin; mannitol; and sorbitol. The amount of diluent(s) in a systemic or topical composition is typically about 50 to about 90% by weight of the composition.

Suitable lubricants include silica, talc, stearic acid and its magnesium salts and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycol and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of theobroma. The amount of lubricant(s) in a systemic or topical composition is typically about 5 to about 10% by weight of the composition.

Suitable binders include polyvinyl pyrrolidone; magnesium aluminum silicate; starches such as com starch and potato starch; gelatin; tragacanth; and cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, methylcellulose, microcrystalline cellulose, and sodium carboxymethylcellulose. The amount of binder(s) in a systemic composition is typically about 5 to about 50% by weight of the composition.

Suitable disintegrants include agar, alginic acid and the sodium salt thereof, effervescent mixtures, croscarmellose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays, and ion exchange resins. The amount of disintegrant(s) in a systemic or topical composition is typically about 0.1 to about 10% by weight of the composition.

Suitable colorants include a colorant such as an FD&C dye. When used, the amount of colorant in a systemic or topical composition is typically about 0.005 to about 0.1% by weight of the composition.

Suitable flavors include menthol, peppermint, and fruit flavors. The amount of flavor(s), when used, in a systemic or topical composition is typically about 0.1 to about 1.0%.

Suitable sweeteners include aspartame and saccharin. The amount of sweetener(s), when used, in a systemic or topical composition is typically about 0.001 to about 1% by weight of the composition.

Suitable antioxidants include butylated hydroxyanisole (“BHA”), butylated hydroxytoluene (“BHT”), and vitamin E. The amount of antioxidant(s) in a systemic or topical composition is typically about 0.1 to about 5% by weight of the composition.

Suitable preservatives include benzalkonium chloride, methyl paraben, and sodium benzoate. The amount of preservative(s) in a systemic or topical composition is typically about 0.01 to about 5% by weight of the composition.

Suitable glidants include silicon dioxide. The amount of glidant(s) in a systemic or topical composition is typically about 1 to about 5% by weight of the composition.

Suitable solvents include water, isotonic saline, ethyl oleate, glycerin, hydroxylated castor oils, alcohols such as ethanol, and phosphate buffer solutions. The amount of solvent(s) in a systemic or topical composition is typically from about 0 to about 100% by weight of the composition.

Suitable suspending agents include AVICEL RC-591 (from FMC Corporation of Philadelphia, PA) and sodium alginate. The amount of suspending agent(s) in a systemic or topical composition is typically about 1 to about 8% by weight of the composition.

Suitable surfactants include lecithin, Polysorbate 80, and sodium lauryl sulfate, and the TWEENS from Atlas Powder Company of Wilmington, Delaware. Suitable surfactants include those disclosed in the C.T.F.A. Cosmetic Ingredient Handbook, 1992, pp.587-592; Remington’s Pharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon’s Volume 1, Emulsifiers & Detergents, 1994, North American Edition, pp. 236-239. The amount of surfactant(s) in the systemic or topical composition is typically about 0.1 % to about 5% by weight of the composition.

Although the amounts of components in the systemic compositions may vary depending on the type of systemic composition prepared, in general, systemic compositions include 0.01% to 50% by weight of an active compound and 50% to 99.99% by weight of one or more carriers. Compositions for parenteral administration typically include 0.1% to 10% by weight of actives and 90% to 99.9% by weight of a carrier including a diluent and a solvent.

Compositions for oral administration can have various dosage forms. For example, solid forms include tablets, capsules, granules, and bulk powders. These oral dosage forms include a safe and effective amount, usually at least about 5% by weight, and more particularly from about 25% to about 50% by weight of actives. The oral dosage compositions include about 50% to about 95% by weight of carriers, and more particularly, from about 50% to about 75% by weight.

Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated, or multiple-compressed. Tablets typically include an active component, and a carrier comprising ingredients selected from diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, glidants, and combinations thereof. Specific diluents include calcium carbonate, sodium carbonate, mannitol, lactose and cellulose. Specific binders include starch, gelatin, and sucrose. Specific disintegrants include alginic acid and croscarmellose. Specific lubricants include magnesium stearate, stearic acid, and talc. Specific colorants are the FD&C dyes, which can be added for appearance. Chewable tablets preferably contain sweeteners such as aspartame and saccharin, or flavors such as menthol, peppermint, fruit flavors, or a combination thereof.

Capsules (including implants, time release and sustained release formulations) typically include an active compound (e.g., a compound of formula (I)), and a carrier including one or more diluents disclosed above in a capsule comprising gelatin. Granules typically comprise a disclosed compound, and preferably glidants such as silicon dioxide to improve flow characteristics. Implants can be of the biodegradable or the non-biodegradable type.

The selection of ingredients in the carrier for oral compositions depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of this disclosure. Solid compositions may be coated by conventional methods, typically with pH or timedependent coatings, such that a disclosed compound is released in the gastrointestinal tract in the vicinity of the desired application, or at various points and times to extend the desired action. The coatings typically include one or more components selected from the group consisting of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, EUDRAGIT® coatings (available from Evonik Industries of Essen, Germany), waxes and shellac.

Compositions for oral administration can have liquid forms. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted from non- effervescent granules, suspensions reconstituted from non-effervescent granules, effervescent preparations reconstituted from effervescent granules, elixirs, tinctures, syrups, and the like. Liquid orally administered compositions typically include a disclosed compound and a carrier, namely, a carrier selected from diluents, colorants, flavors, sweeteners, preservatives, solvents, suspending agents, and surfactants. Peroral liquid compositions preferably include one or more ingredients selected from colorants, flavors, and sweeteners.

Other compositions useful for attaining systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically include one or more of soluble filler substances such as diluents including sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose, and hydroxypropyl methylcellulose. Such compositions may further include lubricants, colorants, flavors, sweeteners, antioxidants, and glidants.

The disclosed compounds can be topically administered. Topical compositions that can be applied locally to the skin may be in any form including solids, solutions, oils, creams, ointments, gels, lotions, shampoos, leave-on and rinse-out hair conditioners, milks, cleansers, moisturizers, sprays, skin patches, and the like. Topical compositions include: a disclosed compound (e.g., a compound of formula (I)), or a pharmaceutically acceptable salt thereof), and a carrier. The carrier of the topical composition preferably aids penetration of the compounds into the skin. The carrier may further include one or more optional components.

The amount of the carrier employed in conjunction with a disclosed compound is sufficient to provide a practical quantity of composition for administration per unit dose of the compound. Techniques and compositions for making dosage forms useful in the methods of this disclosure are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, cds. (1979); Lieberman ct al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd Ed., (1976).

A carrier may include a single ingredient or a combination of two or more ingredients. In the topical compositions, the carrier includes a topical carrier. Suitable topical carriers include one or more ingredients selected from phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, symmetrical alcohols, aloe vera gel, allantoin, glycerin, vitamin A and E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbide, castor oil, combinations thereof, and the like. More particularly, carriers for skin applications include propylene glycol, dimethyl isosorbide, and water, and even more particularly, phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, and symmetrical alcohols.

The carrier of a topical composition may further include one or more ingredients selected from emollients, propellants, solvents, humectants, thickeners, powders, fragrances, pigments, and preservatives, all of which are optional.

Suitable emollients include stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane- 1,2-diol, butane- 1,3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, arachis oil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof. Specific emollients for skin include stearyl alcohol and polydimethylsiloxane. The amount of emollient(s) in a skin-based topical composition is typically about 5% to about 95% by weight of the composition.

Suitable propellants include propane, butane, isobutane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof. The amount of propellant(s) in a topical composition is typically about 0% to about 95% by weight of the composition.

Suitable solvents include water, ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethylsulfoxide, dimethyl formamide, tetrahydrofuran, and combinations thereof. Specific solvents include ethyl alcohol and homotopic alcohols. The amount of solvent(s) in a topical composition is typically about 0% to about 95% by weight of the composition.

Suitable humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. Specific humectants include glycerin. The amount of humectant(s) in a topical composition is typically 0% to 95% by weight of the composition.

The amount of thickener(s) in a topical composition is typically about 0% to about 95% by weight of the composition.

Suitable powders include beta-cyclodextrins, hydroxypropyl cyclodextrins, chalk, talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide, sodium polyacrylate, tetra alkyl ammonium smectites, trialkyl aryl ammonium smectites, chemically-modified magnesium aluminum silicate, organically-modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and combinations thereof. The amount of powder(s) in a topical composition is typically 0% to 95% by weight of the composition.

The amount of fragrance in a topical composition is typically about 0% to about 0.5%, particularly, about 0.001% to about 0.1% by weight of the composition.

Suitable pH adjusting additives include HC1 or NaOH in amounts sufficient to adjust the pH of a topical pharmaceutical composition.

Methods of Use

The disclosed compounds and pharmaceutical compositions may be used in methods for treatment of disorders, such as a disorder characterized or mediated by the Y220C mutant of p53. In some embodiments, the disclosed compounds and pharmaceutical compositions are useful in methods of treating proliferative disorders such as cancers.

Accordingly, in some embodiments, disclosed herein is a method of treating a disorder in a subject in need thereof, wherein the disorder is characterized or mediated by the Y220C mutant of p53, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein (e.g., a compound of formula (I)), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of formula (I)), or a pharmaceutically acceptable salt thereof. In some embodiments, the disorder is a proliferative disease, i.c., a disease that occurs due to abnormal growth or extension by the multiplication of cells. In some embodiments, the proliferative disease is cancer. The term “cancer” refers to a class of diseases characterized by development of abnormal cells that proliferate uncontrollably and have the ability to infiltrate and destroy normal body tissues. See, e.g., Stedman’s Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990.

In some embodiments, the compounds and pharmaceutical compositions disclosed herein are used for treating cancer in a subject in need thereof. In some embodiments, the cancer is a cancer in which expression of one or more p53 target genes is reduced relative to non-cancerous cells. In some embodiments, the cancer is a solid tumor, such as a sarcoma or a carcinoma. In some embodiments, the cancer is a hematologic malignancy.

Exemplary sarcomas include, but are not limited to, alveolar rhabdomyosarcoma, alveolar soft part sarcoma, ameloblastoma, angiosarcoma, chondrosarcoma, chordoma, clear cell sarcoma of soft tissue, dedifferentiated liposarcoma, desmoid, desmoplastic small round cell tumor, embryonal rhabdomyosarcoma, epithelioid fibrosarcoma, epithelioid hemangioendothelioma, epithelioid sarcoma, esthesioneuroblastoma, Ewing sarcoma, extrarenal rhabdoid tumor, extraskeletal myxoid chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, giant cell tumor, hemangiopericytoma, infantile fibrosarcoma, inflammatory myofibroblastic tumor, Kaposi sarcoma, leiomyosarcoma of bone, liposarcoma, liposarcoma of bone, malignant fibrous histiocytoma (MEH), malignant fibrous histiocytoma (MFH) of bone, malignant mesenchymoma, malignant peripheral nerve sheath tumor, mesenchymal chondrosarcoma, myxofibrosarcoma, myxoid liposarcoma, myxoinflammatory fibroblastic sarcoma, neoplasms with perivascular epithelioid cell differentiation, osteosarcoma, parosteal osteosarcoma, neoplasm with perivascular epithelioid cell differentiation, periosteal osteosarcoma, pleomorphic liposarcoma, pleomorphic rhabdomyosarcoma, PNET/extraskeletal Ewing tumor, rhabdomyosarcoma, round cell liposarcoma, small cell osteosarcoma, solitary fibrous tumor, synovial sarcoma, and telangiectatic osteosarcoma.

Exemplary carcinomas include, but are not limited to, adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, anaplastic carcinoma, large cell carcinoma, small cell carcinoma, anal cancer, appendix cancer, bile duct cancer (i.e., cholangiocarcinoma), bladder cancer, brain tumor, breast cancer, cervical cancer, colon cancer, cancer of unknown primary (CUP), esophageal cancer (c.g., esophageal squamous cell carcinoma), eye cancer, fallopian tube cancer, gastroenterological cancer, kidney cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer, medulloblastoma, melanoma, oral cancer, ovarian cancer, pancreatic cancer, parathyroid disease, penile cancer, pituitary tumor, prostate cancer, rectal cancer, skin cancer, stomach cancer, testicular cancer, throat cancer, thyroid cancer, uterine cancer, vaginal cancer, and vulvar cancer.

Exemplary hematologic malignancies include, but are not limited to, leukemias, lymphomas, myelomas, non-Hodgkin’s lymphomas, Hodgkin’s lymphomas, T-cell malignancies, and B-cell malignancies. Exemplary T-cell malignancies include anaplastic large cell lymphoma, angioimmunoblastic lymphoma, adult T-cell leukemia/lymphoma (ATLL), blastic NK-cell lymphoma, cutaneous T-cell lymphoma, enteropathy-type T-cell lymphoma, hematosplenic gamma-delta T-cell lymphoma, lymphoblastic lymphoma, nasal NK/T-cell lymphomas, peripheral T-cell lymphoma not otherwise specified (PTCL-NOS), and treatment- related T-cell lymphomas. Exemplary B-cell malignancies include chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), high risk CLL, and a non-CLL/SLL lymphoma. In some embodiments, the cancer is selected from B cell prolymphocytic leukemia, Burkitt’s lymphoma, diffuse large B-cell lymphoma (DLBCL), extranodal marginal zone B cell lymphoma, follicular lymphoma (FL), immunoblastic large cell lymphoma, intravascular large B cell lymphoma, lymphomatoid granulomatosis, lymphoplasmacytic lymphoma, mantle cell lymphoma (MCL), mediastinal (thymic) large B cell lymphoma, multiple myeloma, nodal marginal zone B cell lymphoma, non-Burkitt high grade B cell lymphoma, plasma cell myeloma, plasmacytoma, precursor B-lymphoblastic lymphoma, primary effusion lymphoma, primary mediastinal B-cell lymphoma (PMBL), splenic marginal zone lymphoma, or Waldenstrom’s macroglobulinemia.

In some embodiments, the cancer is selected from thyroid cancer, lung cancer, breast cancer, liver cancer, stomach cancer, pancreatic cancer, colorectal cancer, kidney cancer, urethral cancer, ovarian cancer, prostate cancer, bone cancer, bladder cancer, esophageal cancer, brain cancer, head and neck cancer, skin cancer, a lymphoma, a leukemia, and multiple myeloma. In some embodiments, the cancer is a relapsed or refractory cancer, such as a cancer described herein. In some embodiments, the cancer is a metastasized cancer, such as a cancer described herein.

In the methods of treatment disclosed herein, a compound or pharmaceutical composition may be administered to the subject by any convenient route of administration, whether systemically/peripherally or at the site of desired action, including but not limited to, oral (e.g., by ingestion); topical (including e.g. transdermal, intranasal, ocular, buccal, and sublingual); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., an aerosol, e.g., through mouth or nose); rectal; vaginal; parenteral (e.g., by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal injection); or by implant of a depot, for example, subcutaneously or intramuscularly. In some embodiments, the administration comprises oral administration. In some embodiments, the administration comprises parenteral administration. Additional modes of administration may include adding the compound and/or a composition comprising the compound to a food or beverage, including a water supply for an animal, to supply the compound as part of the animal’s diet.

It will be appreciated that appropriate dosages of the compounds, and compositions comprising the compounds, can vary from patient to patient. Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleterious side effects of the treatments of the present disclosure. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, and/or materials used in combination, and the age, sex, weight, condition, general health, and prior medical history of the patient. The amount of compound and route of administration will ultimately be at the discretion of the physician, although generally the dosage will be to achieve local concentrations at the site of action which achieve the desired effect without causing substantial harmful or deleterious side-effects.

Administration in vivo can be effected in one dose, continuously or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician. In general, a suitable dose of the compound is in the range of about 100 p.g to about 250 mg per kilogram body weight of the subject per day.

The compound or composition may be administered once, on a continuous basis (e.g. by an intravenous drip), or on a periodic/intermittent basis, including about once per hour, about once per two hours, about once per four hours, about once per eight hours, about once per twelve hours, about once per day, about once per two days, about once per three days, about twice per week, about once per week, and about once per month. The composition may be administered until a desired reduction of symptoms is achieved.

A compound described herein may be used in combination with other known therapies. Administered “in combination,” as used herein, means that two (or more) different treatments are delivered to the subject during the course of the subject's affliction with the disorder, e.g., the two or more treatments are delivered after the subject has been diagnosed with the disorder and before the disorder has been cured or eliminated or treatment has ceased for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery.” In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered. A compound or composition described herein and the at least one additional therapeutic agent can be administered simultaneously, in the same or in separate compositions, or sequentially. For sequential administration, the compound described herein can be administered first, and the additional agent can be administered subsequently, or the order of administration can be reversed.

In some embodiments, a compound described herein is administered in combination with other therapeutic treatment modalities, including surgery, radiation, transplantation (e.g., stem cell transplantation, bone marrow transplantation), cryotherapy, and/or thermotherapy. Such combination therapies may allow for lower dosages of the administered agent and/or other chemotherapeutic agent, thus avoiding possible toxicities or complications associated with the various therapies.

In some embodiments, the compound described herein is administered with at least one additional therapeutic agent, such as a chemotherapeutic agent. In certain embodiments, the compound described herein is administered in combination with one or more additional chemotherapeutic agents. The chemotherapeutic agent may be a chemotherapeutic agent identified on the “A to Z List of Cancer Drugs” published by the National Cancer Institute.

Also disclosed herein are methods of reducing proliferation of cancer cells in a sample, comprising contacting the sample with a compound described herein (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof), or a pharmaceutical composition described herein.

Kits

Compounds and/or compositions disclosed herein may be assembled into kits or pharmaceutical systems. Kits or pharmaceutical systems according may include a carrier or package such as a box, carton, tube or the like, having in close confinement therein one or more containers, such as vials, tubes, ampoules, or bottles, which contain a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. Kits or pharmaceutical systems may also include printed instructions for using the compounds and/or compositions.

The following examples further illustrate aspects of the disclosure, but should not be construed as in any way limiting its scope. EXAMPLES

The following abbreviations are used in the Examples: ACN is acetonitrile; AcOH is acetic acid; DCM is dichloromethane; DIPEA is ALV-diisopropylcthylaminc; DMF is N,N- dimethylformamide; DMSO is A-di methyl sulfoxide; EA is ethyl acetate; HATU is 1- [bis(dimethylamino)methylene]- 1 H- 1 ,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate; Hex is hexanes; LAH is lithium aluminum hydride; MeOH is methanol; TEA is triethylamine; TFA is trifluoroacetic acid; THF is tetrahydrofuran;

Example 1: Compound Syntheses

Procedure A: Preparation of JQ-1 amine linker

To a solution of (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2/][l,2,4]triazolo[4,3-«][l,4]diazepin-6-yl)acetic acid 1 (50 mg, 0.12 mmol, 1 eq.) and tert-butyl (4-aminobutyl)carbamate (23 mg, 0.12 mmol, 1 eq.) in DCM (1 mL) was added HATU (52 mg, 0.14 mmol, 1.1 eq.) and DIPEA (65 pL, 0.37 mmol, 3 eq.), and the reaction mixture was stirred at room temperature for 1 hour. The crude mixture was purified by flash chromatography with 0-10% MeOH/DCM to afford the coupling product as a colorless oil. The product was then dissolved in DCM (5 mL) and subjected to TFA (1 mL) at room temperature for 1 hour. The solvent was then evaporated to afford compound 2 as a yellow oil (55 mg, 94% in two steps) which will be used directly in the next step.

Procedure B : of the

A solution of 2-Iodo-4-nitro-lH-indole 3 (403 mg, 1.74 mmol, 1 eq.) in DMF (3 mL) was added NaH (60% in oil, 208 mg, 5.21 mmol, 3 eq.) at 0 °C in portions. The solution was then stirred at the same temperature for 1 hour before the addition of 2,2,2-trifluoroethyl trifluoromethylsulfonate (750 pL, 5.22 mmol, 3 eq.). The temperature was then raised to 70 °C and the reaction was stirred for 2 hours. The crude mixture was purified by Cl 8 chromatography column with 10-100% ACN/H2O to afford intermediate 4 as a white solid (470 mg, 73.2%).

Intermediate 4 (470 mg, 1.27 mmol, 1 eq.) and iron (426 mg, 7.26 mmol, 6 eq.) were dissolved in acetic acid (7 mL), and the reaction mixture was stirred for 4 hours at 50 °C. The mixture was then filtered, and the filtrate was concentrated under vacuum and purified by C18 chromatography column with 10-100% ACN/ H2O to afford intermediate 5 a brown solid (410 mg, 95%).

To a solution of intermediate 5 (100 mg, 294 pmol, 1 eq.) and tert-butyl 4-oxopiperidine- 1-carboxylate (211 mg, 1.06 mmol, 3.6 eq.) in methanol (5 mL) was added sodium cyanoborohydride (92.4 mg, 1.47 mmol, 5 eq.). Acetic acid was then added to tune the pH to 5 before stirring at 50 °C overnight. The mixture was concentrated under vacuum and then purified by C18 chromatography column with 10-100% ACN/H2O to afford intermediate 6 (77 mg, 50%) as a colorless oil.

To a solution of (4-aminophenyl)dimethylphosphine oxide 7 (100 mg, 591 pmol, 1 eq.) and 3-bromoprop-l-yne (80% in toluene, 72.4 pL, 650 pmol, 1.1 eq.) in DMF (3 mL) was added potassium carbonate (163 mg, 1.18 mmol, 2 eq.), and the reaction mixture was stirred at 100 °C for 1 hour. The crude was then purified by Cl 8 chromatography column with 10-100% ACN/H2O to afford intermediate 8 (65.6 mg, 53.5%) as a yellow oil.

A solution of intermediate 6 (12 mg, 23 pmol, 1 eq.), intermediate 8 (4.7 mg, 23 pmol, 1 eq.), bis(triphenylphosphine)palladium(II) dichloride (1.6 mg, 2.3 pmol, 0.1 eq.), and copper(I) iodide (0.22 mg, 1.1 pmol, 0.05 eq.) in THF (1 mL) under nitrogen was added triethylamine (1 mL), and the reaction was stirred at room temperature for 1 hour. The crude was then purified by HPLC with MeOH/H2O (0.035% TFA) to give the intermediate 9 as a brown solid (3.04 mg, 22%).

The intermediate 9 (200 mg, 332 pmol, 1 eq.) was dissolved in DCM (5 mL) and subjected to TFA (1 mL), and the reaction mixture was stirred for 2 hours before concentrating under reduced pressure. The crude was then dissolved in ACN (3 mL) followed by the addition of potassium carbonate (138 mg, 997 pmol, 3 eq.) and tert-butyl 2-bromoacetate (146 pL, 997 pmol, 3 eq.). After stirring for 2 hours at room temperature, the reaction mixture was purified by C18 chromatography column with 10-100% ACN/H2O to afford the reaction product. The reaction product was then dissolved in DCM (5 mL) with TFA (1 mL), and the solution was stirred for 2 hours before concentrating under reduced pressure to afford compound 10 in crude (130 mg, 70% in total) for the next step directly.

Procedure C: Preparation of the Compound P444

To a solution of compound 2 (5.0 mg, 11 pmol, 1 eq.), compound 10 (6.0 mg, 11 pmol, 1 eq.), HATU (4.8 mg, 13 pmol, 1.2 eq.) in DMF (200 pL) was added DIPEA (7.4 pL, 53 pmol, 5 eq.), and the reaction was stirred at room temperature overnight. The crude was then purified by HPLC with MCOH/H2O (0.035% TFA) to give compound P444 as a white solid (3.53 mg, 33%). MS obsd. [(M+H)+]: 1013.37. H NMR (500 MHz, d⁶-DMSO) 8 9.76 (s, 1H), 8.55 (d, J = 5.9 Hz, 1H), 8.21 (t, J= 5.7 Hz, 1H), 7.49 (m, 4H), 7.45 - 7.39 (m, 2H), 7.17 - 7.07 (m, 1H), 7.02 (t, 7= 8.0 Hz, 1H), 6.80 (dd, 7= 8.6, 2.2 Hz, 2H), 6.74 (d, 7 = 8.3 Hz, 1H), 6.21 (d, 7 = 7.9 Hz, 1H), 4.94 (q, 7 = 9.4 Hz, 2H), 4.50 (t, 7 = 7.1 Hz, 1H), 4.29 (s, 2H), 3.96 - 3.87 (m, 2H), 3.63 - 3.55 (m, 1H), 3.55 - 3.49 (m, 2H), 3.32 (s, 1H), 3.32 - 3.07 (m, 8H), 2.59 (s, 3H), 2.40 (s, 3H), 2.17 - 2.10 (m, 2H), 1.83 - 1.74 (m, 2H), 1.62 (s, 3H), 1.55 (d, 7= 13.1 Hz, 6H), 1.48 (br, 4H).

Procedure D: Preparation of the Compound 14 and Compound 16

A solution containing 2-bromo-9H-carbazole 11 (2.00 g, 9.13 mmol, 1 eq.) and NaOH (1.40 g, 35 mmol, 4.3 eq.) in DMF (20 mL) was stirred for 30 minutes under room temperature before the addition of iodoethane (1.13 mL, 14.0 mmol, 1.72 eq.). The reaction was stirred overnight at room temperature. The resulting solution was then diluted in water and extracted with ethyl acetate 3 times. The combined organic phase was washed with 5% LiCl aqueous solution 3 times, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude was purified with flash chromatography with 0-10% EA/Hex to afford intermediate 12 as a white solid (2.1 g, 94%).

The intermediate 12 (2.4 g, 8.8 mmol, 1 eq.) was dissolved in DMF (16 mL), and the solution was cooled to 0 °C before the addition of phosphoryl chloride (2.4 mL, 26 mmol, 3 eq.) dropwise. The reaction was then warmed to room temperature and then heated at 70 °C overnight. After cooling to room temperature, the reaction was quenched with ice and neutralized with 5% aqueous sodium hydroxide. The resulting solution was extracted with ethyl acetate 3 times, and the combined organic layer was washed with water and brine, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude was then purified with flash chromatography with 0-10% EA/Hex to afford intermediate 13 as a white solid (1 .90 g, 72%).

A solution of intermediate 13 (800 mg, 2.65 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-(4- methylthiophen-2-yl)-l,3,2-dioxaborolane (890 mg, 3.97 mmol, 1.5 eq.), potassium carbonate (732 mg, 5.30 mmol, 2 eq.), and [l,l'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (194 mg, 265 pmol, 0.1 eq.) in 1,4-dioxane (30 mL) and water (2.65 mL) was stirred at 100 °C overnight under nitrogen. The resulting crude was concentrated under reduced pressure and then purified by flash chromatography 0-10% EA/Hex to afford compound 14 as a white solid (731 mg, 86.4%).

The compound 14 (731 mg, 2.29 mmol, 1 eq.) was dissolved in MeOH (30 mL) followed by the addition of hydroxylamine hydrochloride (318 mg, 4.58 mmol, 2 eq.), sodium acetate (375 mg, 4.58 mmol, 2 eq.), and water (3.3 mL). The solution was stirred at room temperature for 2 hours and was then diluted with ethyl acetate. The resulting solution was washed with water, and the aqueous phase was extracted twice with ethyl acetate. The combined organic phase was then dried over anhydrous sodium sulfate and concentrated under reduced pressure to afford compound 15 (557 mg, 72.8%) which was used directly for the next step.

To a solution of compound 15 (557 mg, 1.73 mmol, 1 eq.) in dry THF (6 mL) was added a suspension of LiAlEL (228 mg, 6.00 mmol, 3.5 eq.) in dry THF (6 mL) dropwise. The resulting solution was refluxed for 1 hour before cooling to room temperature and quenched with MeOH. The solution was then filtered and then evaporated under reduced pressure, and the residue was then purified by flash chromatography with 0-20% MeOH/DCM to afford compound 16 as a brown solid (250 mg, 45%).

Procedure E: Preparation of the Compound P408

To a solution of compound 24 (synthesized following Procedure A, 7.8 mg, 16 pmol, 1 eq.) and compound 14 (5.0 mg, 16 pmol, 1 eq.) in MeOH (300 pL) was acetic acid to tune the pH to 5 before the addition of sodium cyanoborohydride (3.0 mg, 47 pmol, 3 eq.). The reaction was stirred overnight at room temperature and was then purified by HPLC with MeOH/HaO (0.035% TFA) to give compound P408 as a white solid (6.76 mg, 54%). MS obsd. [(M+H)+]: 802.30. ^ NMR (500 MHz, d⁶-DMSO) 5 8.75 (s, 2H), 8.24 (d, J = 1.7 Hz, 1H), 8.19 (t, J = 5.7 Hz, 1H), 8.11 (d, 7= 8.1 Hz, 1H), 7.91 (d, J= 1.5 Hz, 1H), 7.70 (d, J= 8.4 Hz, 1H), 7.57 (dd, J = 8.4, 1.7 Hz, 1H), 7.52 (d, 7 = 1.4 Hz, 1H), 7.51 - 7.47 (m, 3H), 7.42 (d, 7= 8.7 Hz, 2H), 7.15 (t, 7= 1.3 Hz, 1H), 4.57 - 4.47 (m, 3H), 4.30 (t, 7 = 5.8 Hz, 2H), 3.24 (d, 7= 7.2 Hz, 2H), 3.12 (hept, 7= 6.4 Hz, 2H), 3.04 - 2.93 (m, 2H), 2.59 (s, 3H), 2.40 (s, 3H), 2.29 (d, 7= 1.1 Hz, 3H), 1.68 - 1.62 (m, 2H), 1.61 (s, 3H), 1.46 (t, 7 = 6.9 Hz, 2H), 1.37 - 1.31 (m, 7H).

Procedure F: Preparation of JQ-1 carboxylic acid linker

1 17

To a solution of (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-677- thieno[3,2/][l,2,4]triazolo[4,3-«][l,4]diazepin-6-yl)acetic acid 1 (40 mg, 0.10 mmol, 1 eq.) and tert-butyl 4-aminobutanoate, HC1 (19.5 mg, 0.10 mmol, 1 eq.) in DCM (1 mL) was added HATU (45.5 mg, 0.12 mmol, 1.2 eq.) and DIPEA (70 pL, 0.40 mmol, 4 eq.), and the reaction mixture was stirred at room temperature for 1 hour. The crude mixture was purified by flash chromatography with 0-10% MeOH/DCM to afford the coupling product as a colorless oil. The product was then dissolved in DCM (5 mL) and subjected to TFA (1 mL) at room temperature for 1 hour. The solvent was then evaporated to afford compound 2 as a yellow oil (50 mg, 92% in two steps) which will be used directly in the next step. Procedure G: Preparation the Compound P009

To a solution of compound 16 (6.6 mg, 21 pmol, 1 cq.), compound 17 (10 mg, 21 pmol, 1 eq.), HATU (9.4 mg, 25 pmol, 1.2 eq.) in DMF (200 pL) was added DIPEA (36 pL, 210 pmol, 10 eq.), and the reaction was stirred at room temperature overnight. The crude was then purified by HPLC with McOH/HiO (0.035% TFA) to give compound P009 as a white solid (8.88 mg, 55%). MS obsd. L(M+H)+J: 788.48. NMR (500 MHz, d⁶-DMSO) 5 8.29 (t, J = 5.8 Hz, 1H), 8.24 (t, J = 5.6 Hz, 1H), 8.10 (d, 7= 8.1 Hz, 1H), 7.97 (d, 7= 1.7 Hz, 1H), 7.82 (d, 7= 1.5 Hz, 1H), 7.53 (d, 7= 8.4 Hz, 1H), 7.48 (d, 7 = 1.4 Hz, 1H), 7.46 (d, 7 = 8.8 Hz, 2H), 7.41 (d, 7 = 8.7 Hz, 2H), 7.38 (dd, 7 = 8.1, 1.5 Hz, 1H), 7.35 (dd, 7 = 8.4, 1.7 Hz, 1H), 7.12 (q, 7 = 1.2 Hz, 1H), 4.53 (dd, 7= 8.1, 6.3 Hz, 1H), 4.51 - 4.37 (m, 4H), 3.31 - 3.08 (m, 4H), 2.55 (s, 3H), 2.35 (d, 7 = 0.9 Hz, 3H), 2.27 (d, 7 = 1.1 Hz, 3H), 2.24 (dd, 7 = 7.5, 4.4 Hz, 2H), 1.83 - 1.69 (m, 2H), 1.56 (d, 7 = 0.9 Hz, 3H), 1.31 (t, 7 = 7.1 Hz, 3H). Procedure H: Preparation of the Compound 23

A solution of iodine (8.31 g, 32.8 mmol, 4 eq.) in 1,4-dioxane (7 mL) and pyridine (7 mL) was stirred at 0 °C for 15 minutes. To this solution was then added 2-hydroxybenzaldehyde (855 pL, 8. 19 mmol, 1 eq.) and was then refluxed for 4 hours. The resulting solution was then concentrated under reduced pressure. A 20% sodium thiosulfate aqueous solution was then added until the dark residue turned yellow. The solution was then extracted with DCM three times, and the combined organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford intermediate 19 (2.96 g, 96.7%) which was used directly for the next step.

The intermediate 19 (500 mg, 1.34 mmol, 1 eq.), tert-butyl piperidine-4-ylcarbamate (268 mg, 1.34 mmol, 1 eq.), and acetic acid (76.5 pL, 1.34 mmol, 1 eq.) were dissolved in DCM (5 mL) followed by the addition of sodium triacetoxyborohydride (326 mg, 1.54 mmol, 1.15 eq.). After stirring for overnight, the solution was diluted with DCM, washed with saturated sodium bicarbonate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford intermediate 20 (737 mg, 96.3%) as crude.

To a solution of aniline 21 (980 pL, 10.7 mmol, 1 eq.) in DMF (1 mL) was added potassium carbonate (742 mg, 5.37 mmol, 0.5 eq.). The resulting solution was stirred at room temperature for 5 minutes and was then added 3-bromoprop-l-yne (203 pL, 2.68 mmol, 0.25 eq.) dropwise. After stirring overnight at room temperature, the mixture was diluted with water and extracted with ethyl acetate three times. The combined organic layer was then washed with water and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was then purified by flash chromatography with 0-10% EA/Hex to afford intermediate 22 as a yellow oil (303 mg, 21.5%)

To a solution of intermediate 20 (737 mg, 1.32 mmol, 1 eq.), intermediate 22 (173 mg, 1.32 mmol, 1 eq.), Cui (50.3 mg, 264 pmol, 0.2 eq.), and tetrakis(triphenylphosphine)palladium(0) (76.3 mg, 66 pmol, 0.05 eq.) in THF (13.2 mL) was added triethylamine (460 pL, 3.30 mmol, 2.5 eq.) under nitrogen. The solution was stirred at room temperature overnight. The resulting solution was then diluted with ethyl acetate and washed with saturated sodium bicarbonate solution twice. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was then purified by flash chromatography with 0-10% MeOH/DCM to afford the coupling product. The product was then dissolved in DCM (5 mL) and subjected to TFA (1 mL) at room temperature for 1 hour. The solvent was then removed, and the crude was purified by C 18 chromatography column with 10- 100% ACN/H?O to afford compound 23 (410 mg, 65.3% over two steps) as a light yellow solid. , , , ,

7.4 Hz, 1H), 7.80 (d, 7= 2.0 Hz, 1H), 7.52 - 7.37 (m, 5H), 7.16 - 7.09 (m, 2H), 6.71 - 6.64 (m, 2H), 6.61 (tt, 7 = 7.3, 1.1 Hz, 1H), 4.50 (t, 7 = 7.1 Hz, 1H), 4.24 (s, 2H), 4.10 (s, 2H), 3.77 (s, 1H), 3.28 - 3.37 (m, 2H), 3.29 - 3.16 (m, 3H), 3.15 - 3.05 (m, 4H), 2.60 (s, 3H), 2.40 (s, 3H), 2.11 (br, 2H), 1.90 (br, 2H), 1.63 - 1.71 (m, 2H), 1.61 (s, 3H), 1.49 - 1.59 (m, 4H).

Additional Compounds

Additional compounds were prepared according to synthetic procedures similar to those described above for compounds P444, P407, P009, and P005, using appropriate starting materials. Structures and MS data are shown in Table 1.

Table 1. Compound structures and MS data

Example 2: Biological Data

Materials and Methods

Cell Culture. Human pancreatic cancer (BxPC-3), and a kidney epithelial (HEK293T) cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, V A, USA). Cells were cultured in medium (RPMI 1640 for BxPC-3 cells; DMEM medium for HEK293T cells) supplemented with 10% heat-inactivated FBS, 100 units/mL penicillin, 100 pg/mL streptomycin, and 0.25 pg/mL amphotericin B. Cells were incubated at 37 °C with 5% CO2 in a humidified atmosphere. p53 Y220C Reporter Assay. p53 luciferase reporter lenti virus (BPS Bioscience, San Diego, CA, USA) was used to transduce BxPC-3 cells cultured as described above, and tranductants were selected by growth in 1 pg/mL puromycin (Gibco Invitrogen Corp., Grand Island, NY, USA) added directly to the culture medium. Briefly, cells were seeded in 384-well plates and incubated overnight. Subsequently, the cells were treated with the indicated concentrations of compounds. After 24 h, the plates were subjected to Bright-Glo Luciferase Assay System (Promega, Madison, WI, USA) as described in manufacturer's manual. The proliferation assays were performed in biological triplicate.

Cell viability assay (CellTiter-Glo assay). Cell viability was evaluated using the CellTiter- Glo assay (Promega). Briefly, cells were seeded in 384-well plates and incubated overnight. Subsequently, the cells were treated with the indicated concentrations of compounds. After 72 h, the plates were subjected to CellTiter-Glo as described in manufacturer's manual. The proliferation assays were performed in biological triplicate. IC50 values were determined using a non-linear regression curve fit in GraphPad PRISM 9.5.1.

Western blotting analysis. Total cells lysates were prepared in 2x sample loading buffer (i.e., 250 mM Tris-hydrochloride: pH 6.8, 4% sodium dodecyl sulfate, 10% glycerol, 0.006% bromophenol blue, 2% P-mercaptoethanol, 50 mM sodium fluoride, and 5 mM sodium orthovanadatc). The samples with cell lysates were boiled for 5-8 min at 95 °C. The protein concentrations of the cell lysates were quantified using the BCA method and a BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts of protein were subjected to 4-20% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (Millipore, Bedford, MA, USA) activated with 100% methanol. The membranes were blocked using Intercept® (TBS) Blocking Buffer (LI-COR Biosciences, Lincoln, NE, USA), and subsequently probed with appropriate primary at 4 °C overnight and then incubated with IRDye 800-labeled goat anti-rabbit IgG (LI-COR Biosciences, cat. no. 926-32211) or IRDye 68ORD goat anti-Mouse IgG (LI-COR Biosciences, cat. no. 926- 68070) secondary antibodies at room temperature for 1 hour. After washing the membranes with PBS for 30 min, the membranes were detected on Li-COR Odyssey CLx system.

Co-Immunoprecipitation. HEK293T cells were seeded into 6-well plate (3 x 10⁵ cells/well), cultured overnight, and transfected with 1.5 pg FLAG-tagged full-length BRD4 and 1.5 pg V5-tagged p53 Y220C plasmids using TransIT-LTl transfection reagents (Minis Bio, Madison, WI, USA). The transfected cells were cultured for another 48 h and treated with either compound or DMSO for 4 h before collection. The cells were collected and lysed in Pierce IP Lysis Buffer (Thermo Fisher Scientific) with cOmplete Mini Protease Inhibitor Cocktail (Roche, Basel, Switzerland) for 30 min on ice and centrifuged for 30 min at 4 °C to remove the insoluble fraction. For immunoprecipitation, 20 pL of pre-cleaned anti-FLAG M2 magnetic beads (Sigma- Aldrich, St. Louis, MO, USA) were added to the lysates. The beads-lysate mix was incubated at 4 °C for overnight on a rotator. Beads were magnetically removed and washed three times with PBS, and the FLAG-Tagged protein was competitively eluted using 3X FLAG Peptide (ApexBio Technology, Houston, TX, USA). Immunoblotting was carried out as previously described.

Results

Results of the above experiments are illustrated in FIGS. 2-8. As illustrated in FIG. 2, the bivalent compounds effectively upregulated the p53 reporter. Therefore, it is expected that these bivalent compounds can be used as p53 transcriptional activators in cancer cells. As illustrated in FIG. 3, the bivalent compounds of the present disclosure effectively suppressed the growth of cancer cells. As illustrated in FIG. 4, it the bivalent compounds of the present disclosure effectively induce proximity between BRD4 and the p53 Y220C mutant. As illustrated in FIG. 5, it was confirmed that the bivalent compounds specifically uprcgulatcd the expression of p53 target genes (p21 , MDM2, Puma), and this effect was significantly stronger compared to each individual protein binder. As illustrated in FIG. 6, the regulation of p21 by bivalent compounds occurs within 2 hours. As illustrated in FIG. 7, even after the removal of the bivalent compounds following short-term treatment, their inhibitory effect on cell growth persists. Finally, as illustrated in FIG. 8, modified bivalent compounds designed to bind only to one side of the protein (i.e., negative control compounds), lost their ability to activate the p53 reporter. The structures of the negative control compounds are shown below:

All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

Claims

CLAIMS:

1. A compound of formula (I)

BB-L-PB (I) or a pharmaceutically acceptable salt thereof, wherein: BB is moiety that binds to BRD4;

L is a linker; and

PB is a moiety that binds to a Y220C mutant of p53.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein BB has a formula selected from: 3. The compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, wherein BB is a moiety of formula:

4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein PB is a moiety of formula:

5. The compound of any one of claims 1 -4, or a pharmaceutically acceptable salt thereof, wherein PB is a moiety of formula:

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula (la):

7. The compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein L is a direct bond or comprises any combination of -CH2-, -CH=CH-, -C=C-, -O-, -NR'-, -BR'-, -S-, -C(O)-, -C(NR')-, -S(O)-, -S(O)2-, arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties, wherein the arylene, heteroarylene, cycloalkylene, and heterocyclylene moieties are independently unsubstituted or substituted with 1, 2, or 3 substituents.

8. The compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein L is a direct bond or comprises any combination of -CH2-, -O-, -C(O)-, -NH-, -N(CH3)-, cycloalkylene, and heterocyclylene moieties.

9. The compound of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein L comprises any combination of the following moieties: wherein p is 1, 2, 3, 4, 5, or 6; and q is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein L has a formula selected from:

5 11. The compound of claim 1, wherein the compound is selected from:

and pharmaceutically acceptable salts thereof.

12. A pharmaceutical composition comprising a compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

13. A method of treating a proliferative disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof.

14. The method of claim 13, wherein the proliferative disease is a cancer selected from a carcinoma, a sarcoma, and a hematologic malignancy.

15. The method of claim 14, wherein the cancer is selected from thyroid cancer, lung cancer, breast cancer, liver cancer, stomach cancer, pancreatic cancer, colorectal cancer, kidney cancer, urethral cancer, ovarian cancer, prostate cancer, bone cancer, bladder cancer, esophageal cancer, brain cancer, head and neck cancer, skin cancer, a lymphoma, a leukemia, and multiple myeloma.

16. A compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof, for use as a medicament.

17. A compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof, for use in treating a proliferative disease.

18. The compound of claim 17, wherein the proliferative disease is a cancer selected from a carcinoma, a sarcoma, and a hematologic malignancy.

19. The compound of claim 18, wherein the cancer is selected from thyroid cancer, lung cancer, breast cancer, liver cancer, stomach cancer, pancreatic cancer, colorectal cancer, kidney cancer, urethral cancer, ovarian cancer, prostate cancer, bone cancer, bladder cancer, esophageal cancer, brain cancer, head and neck cancer, skin cancer, a lymphoma, a leukemia, and multiple myeloma.

20. A kit comprising a compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof.