Degradation agent of cyclin-dependent kinase 7 (CDK7) and use thereof

Abstract

Disclosed are bispecific compounds (degraders) that target CDK7 for degradation, as well as pharmaceutical compositions containing the degraders and methods for treating diseases using the compounds.

Description

[0001] Related Applications

[0002] This application claims priority under 35 USC §119(e) to U.S. Provisional Application No. 62 / 882,958, filed on August 5, 2019, the entire contents of which are incorporated herein by reference.

[0003] Government licensing rights

[0004] This invention was made with Government support under Grant No. W81XWH-16-1-0252 awarded by the U.S. Army Medical Research and Development Command. The Government has certain rights in this invention. Background Art

[0005] Cyclin-dependent kinase 7 (CDK7) is a major regulator of cell cycle progression and gene transcription. It has been reported that CDK7 inhibition reduces proliferation and increases cell death in different tumor models (Kwiatkowski et al., Nature 511(7511):616-620(2014); Olson et al., Cell Chem. Biol. 26(6):792-803.e10(2019). Summary of the invention

[0006] The first aspect of the present invention relates to a bispecific compound, which comprises a targeting ligand and a degradation determinant (Degron (D)) covalently linked to each other through a linker, wherein the targeting ligand binds to cyclin-dependent kinase 7 (CDK7), wherein the compound has a structure represented by formula (I):

[0007] in

[0008] R1, R2, R3, R4, R5, L1, L2, A and B are as defined herein, and

[0009] The degron represents the portion that binds the E3 ubiquitin ligase;

[0010] or a pharmaceutically acceptable salt or stereoisomer thereof. The targeting ligand (TL) is linked to the linker (L) via the R2 group of TL.

[0011] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0012] Another aspect of the present invention relates to a process for preparing a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0013] Other aspects of the invention relate to methods of treating a disease or condition involving aberrant (e.g., dysfunctional or dysregulated) CDK7 activity, which requires administering to a subject in need thereof a therapeutically effective amount of a bispecific compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0014] In some embodiments, the disease or condition is cancer.

[0015] In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor is breast cancer, brain cancer, lung cancer, colorectal cancer, neuroblastoma, osteosarcoma, or lymphoma.

[0016] In some embodiments, the cancer is a hematological cancer. In some embodiments, the hematological cancer is a leukemia, lymphoma, or multiple myeloma.

[0017] In some embodiments, the disease or disorder is an autoimmune disease or disorder.

[0018] Without wishing to be bound by any particular theory of operation, the bispecific compounds of formula (I) of the present invention are believed to cause degradation of CDK7 by recruiting the ubiquitin / proteasome system of the cell, which is brought into close proximity to CDK7 due to the binding between CDK7 and the targeting ligand, and the function of the ubiquitin / proteasome system is to routinely recognize and remove damaged proteins. After the CDK7 protein is destroyed, the degrader is released and continues to be active. Applicants have recently discovered a CDK7 inhibitor with low nanomolar potency. By combining this potent CDK7 ligand with an E3 ligase binder, it was found that the bispecific degrader molecules of the present invention are able to recruit E3 ligases and thus promote the degradation of CDK7. Therefore, by engaging and utilizing the body's own natural protein disposal system, the bispecific compounds of the present invention may represent a potential improvement over current CDK7 small molecule inhibitors and may overcome one or more limitations on their use. Therefore, the effective intracellular concentration of the degrader may be significantly lower than that of a small molecule CDK7 inhibitor. In general, the bispecific compounds of the present invention may represent a new set of chemical tools for knocking out CDK7 and may provide a potential treatment for CDK7-related cancers and autoimmune disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1A is an immunoblot showing CDK7 degradation after 6 hours of treatment of Jurkat cells with bispecific compounds 1-10 of the invention and DMSO (negative control).

[0020] Figure 1Bis an immunoblot showing CDK7 degradation after 6 hours of treatment of Jurkat cells with bispecific compounds of the invention 11-20, 3 (positive control) and DMSO (negative control).

[0021] Figure 1C is an immunoblot showing CDK7 degradation after 6 hours of treatment of Jurkat cells with bispecific compounds 21-26 of the invention and DMSO (negative control).

[0022] Figure 2A is an immunoblot showing CDK7 degradation after treatment of Jurkat cells for 2 hours with the selective CDK7 inhibitor YKL-5-124, compound DGY-05-180, the proteasome inhibitor bortezomib, or the acylation inhibitor MLN4924 prior to addition of bispecific compound 3 for 4 hours.

[0023] Figure 2B is an immunoblot showing CDK7 degradation after treatment of Jurkat cells with YKL-5-124, compound DGY-05-180, bortezomib or MLN4924 for 2 hours prior to addition of bispecific compound 20 for 4 hours. DETAILED DESCRIPTION

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the subject matter of this article belongs. As used in the specification and the appended claims, unless otherwise indicated, the following terms have the meanings shown to facilitate understanding of the present invention.

[0025] As used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, "a composition" includes a mixture of two or more such compositions, "an inhibitor" includes a mixture of two or more such inhibitors, and so forth.

[0026] Unless otherwise indicated, the term "about" means within 10% (eg, within 5%, 2%, or 1%) of the particular value modified by the term "about."

[0027] The transitional term "comprising," which is synonymous with "comprising," "including," or "characterized by," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes any elements, steps, or ingredients not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel characteristics of the claimed invention."

[0028] With respect to the compounds of the present invention, to the extent the following terms are used herein to further describe them, the following definitions apply.

[0029] As used herein, the term "alkyl" refers to a saturated straight or branched monovalent hydrocarbon group. In one embodiment, the alkyl group is C1-C 18 In other embodiments, the alkyl group is C0-C6, C0-C5, C0-C3, C1-C 12 , C1-C8, C1-C6, C1-C5, C1-C4 or C1-C3 group (wherein C0 alkyl refers to a bond). Examples of alkyl groups include methyl, ethyl, 1-propyl, 2-propyl, 1-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl. In some embodiments, alkyl groups are C1-C3 alkyl groups. In some embodiments, alkyl groups are C1-C2 alkyl groups.

[0030] As used herein, the term "alkylene" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a group, which is composed only of carbon and hydrogen, does not contain unsaturated bonds and has 1 to 12 carbon atoms, such as methylene, ethylene, propylene, butylene, etc. The alkylene chain can be connected to the rest of the molecule by a single bond and is connected to the group by a single bond. In some embodiments, the alkylene group contains 1 to 8 carbon atoms (C1-C8 alkylene). In other embodiments, the alkylene group contains 1 to 5 carbon atoms (C1-C5 alkylene). In other embodiments, the alkylene group contains 1 to 4 carbon atoms (C1-C4 alkylene). In other embodiments, the alkylene group contains 1 to 3 carbon atoms (C1-C3 alkylene). In other embodiments, the alkylene group contains 1 to 2 carbon atoms (C1-C2 alkylene). In other embodiments, the alkylene group contains one carbon atom (C1 alkylene).

[0031] As used herein, the term "haloalkyl" refers to an alkyl group, as defined herein, substituted with one or more (eg, 1, 2, 3 or 4) halo groups.

[0032] As used herein, the term "alkenyl" refers to a straight or branched monovalent hydrocarbon radical having at least one carbon-carbon double bond. Alkenyl groups include groups having "cis" and "trans" orientations, or "E" and "Z" orientations. In one example, an alkenyl group is C2-C 18 In other embodiments, alkenyl is C2-C 12 、C2-C 10 Examples include ethenyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, but-1,3-dienyl, 2-methylbut-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hex-1,3-dienyl.

[0033] As used herein, the term "alkynyl" refers to a straight or branched monovalent hydrocarbon radical having at least one carbon-carbon triple bond. 18 In other examples, the alkynyl group is a C2-C 12 、C2-C 10 , C2-C8, C2-C6 or C2-C3. Examples include ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl and but-3-ynyl.

[0034] As used herein, the term "alkoxyl" or "alkoxy" refers to an alkyl group as defined above to which an oxygen group is attached. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like. An "ether" is two hydrocarbon groups covalently linked by oxygen. Thus, the substituent of an alkyl group that makes an alkyl group an ether is or is similar to an alkoxy group, and can be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-alkynyl.

[0035] As used herein, the term "halogen" (or "halo" or "halide") refers to fluorine, chlorine, bromine or iodine.

[0036] As used herein, the term "carbocycle" (also referred to as "carbocyclyl") refers to a group used alone or as part of a larger portion, which comprises a saturated, partially unsaturated or aromatic ring system having 3 to 20 carbon atoms, alone or as part of a larger portion (e.g., an alkyl carbocyclic group). The term carbocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged and spirocyclic systems and combinations thereof. In one embodiment, the carbocyclyl includes 3 to 15 carbon atoms (C3-C 15 In one embodiment, the carbocyclyl group comprises 3 to 12 carbon atoms (C3-C 12 In another embodiment, carbocyclyl includes C3-C8, C3-C10 or C5-C 10 In another embodiment, the carbocyclyl as a monocyclic ring includes C3-C8, C3-C6 or C5-C6. In some embodiments, the carbocyclyl as a bicyclic ring includes C7-C 12 In another embodiment, the carbocyclyl group as a spiro ring system includes C5-C 12 Representative examples of monocyclic carbocyclyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, perdeuterated cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, phenyl, and cyclododecyl; bicyclic carbocyclyls having 7 to 12 ring atoms include [4,3], [4,4], [4,5], [5,5], [5,6], or [6,6] ring systems, such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, naphthalene, and bicyclo[3.2.2]nonane. Representative examples of spirocyclic carbocyclyl include spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane and spiro[4.5]decane.The term carbocyclyl includes aryl ring systems defined herein.The term carbocyclyl also includes cycloalkyl rings (e.g., saturated or partially unsaturated single, double or spiral carbocycles).The term carbocyclic group also includes carbocycles fused to one or more (e.g., 1, 2 or 3) different cyclic groups (e.g., aryl or heterocycles), wherein the group or point of attachment is on the carbocyclic ring.

[0037] Thus, the term carbocycle also includes carbocyclylalkyl groups, which, as used herein, refer to groups of the formula --R c -Carbocyclic group, where R c The term carbocycle also includes carbocyclylalkoxy groups, which, as used herein, refer to a carbocyclylalkoxy group of the formula --O--R c - a group bonded to the oxygen atom of a carbocyclic group, wherein R c It is an alkylene chain.

[0038] As used herein, the term "heterocyclyl" refers to a "carbocyclyl" used alone or as part of a larger moiety, comprising a saturated, partially unsaturated or aromatic ring system in which one or more (e.g., 1, 2, 3, or 4) carbon atoms have been replaced by heteroatoms (e.g., O, N, N(O), S, S(O) or S(O)2). The term heterocyclyl includes monocyclic, bicyclic, tricyclic, fused, bridged, and spirocyclic ring systems and combinations thereof. In some embodiments, heterocyclyl refers to a 3- to 15-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a 3- to 12-membered heterocyclyl ring system. In some embodiments, heterocyclyl refers to a saturated ring system, such as a 3- to 12-membered saturated heterocyclyl ring system. In some embodiments, heterocyclyl refers to a heteroaryl ring system, such as a 5- to 14-membered heteroaryl ring system. The term heterocyclyl also includes C3-C8 heterocycloalkyl, which is a saturated or partially unsaturated mono-, bi- or spiro ring system containing 3-8 carbons and one or more (1, 2, 3 or 4) heteroatoms.

[0039] In some embodiments, heterocyclic radical includes 3-12 ring atoms, and includes monocyclic, bicyclic, tricyclic and spirocyclic systems, wherein the ring atoms are carbon, and 1-5 ring atoms are heteroatoms, such as nitrogen, sulfur or oxygen. In some embodiments, heterocyclic radical includes 3- to 7-membered monocycles with one or more heteroatoms selected from nitrogen, sulfur or oxygen. In some embodiments, heterocyclic radical includes 4- to 6-membered monocycles with one or more heteroatoms selected from nitrogen, sulfur or oxygen. In some embodiments, heterocyclic radical includes 3-membered monocycles. In some embodiments, heterocyclic radical includes 4-membered monocycles. In some embodiments, heterocyclic radical includes 5-6-membered monocycles. In some embodiments, heterocyclic radical includes 0 to 3 double bonds. In any of the foregoing embodiments, heterocyclic radical includes 1, 2, 3 or 4 heteroatoms. Any nitrogen or sulfur heteroatom can be optionally oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatom can be optionally quaternized (e.g., [NR4] + Cl - 、[NR4] + OH -Representative examples of heterocyclic groups include oxirane, aziridinyl, thiiridinyl, azetidinyl, oxetanyl, thienyl, 1,2-bithiphenyl, 1,3-bithiphenyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydropyranyl, dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, thiazinanyl, thioxanyl, homopiperazinyl, homopiperidinyl, homopiperidinyl), azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolyl, isothiazolyl, 1,1-dioxothiazolinyl, oxazolidinyl, imidazolidinyl, 4,5-dihydrothiopyranyl, , 6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-dihydroimidazolyl[4,5-d]pyrrolo[2,3-b]pyridinyl, thiazinyl, thienyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, indolyl, thiapyranyl, 2H-pyranyl, 4H-pyranyl, dioxanthenyl, 1,3-dioxolanyl, pyrazolinyl, pyrazolidinyl, dithianyl, dithioanthenyl, pyrimidinylnonyl, pyrimidinyldinonyl, pyrimidine-2,4-dione yl, piperazinylnonyl, piperazinyldinyl, pyrazolidinylimidazolyl, 3-azabicyclo[3.1.0]hexyl, 3,6-diazabicyclo[3.1.1]heptyl, 6-azabicyclo[3.1.1]heptyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 2-azabicyclo[3.2.1]octyl, 8-azabicyclo[3.2.1]octyl, 2-azabicyclo[2.2.2]octyl, 8-azabicyclo[2.2.2]octyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonyl, azaspiro[2.5]octyl, azaspiro[4.5]decyl, 1-azaspiro[4.5]decan-2-yl, azaspiro[5.5] undecyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxane hydropyranyl. Examples of 5-membered heterocycles containing sulfur or oxygen atoms and 1-3 nitrogen atoms are thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl, such as oxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl, and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocyclic rings containing 2-4 nitrogen atoms include imidazolyl, such as imidazol-2-yl; triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl and tetrazolyl, such as 1H-tetrazolyl. Representative examples of benzo-fused 5-membered heterocyclic groups are benzooxazol-2-yl, benzothiazol-2-yl and benzimidazol-2-yl. Examples of 6-membered heterocyclic groups include 1-3 nitrogen atoms and optionally sulfur or oxygen atoms, such as pyridyl, such as pyridin-2-yl, pyridin-3-yl and pyridin-4-yl; pyrimidinyl, such as pyrimidin-2-yl and pyrimidin-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, especially pyridazin-3-yl and pyrazinyl. Pyridine N-oxides and pyridazine N-oxides, as well as pyridyl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl are other examples of heterocyclic groups. In some embodiments, heterocyclic groups include a heterocyclic ring fused to one or more (e.g., 1, 2, or 3) different cyclic groups (e.g., carbocyclic or heterocyclic), wherein the radical or point of attachment is on the heterocyclic ring, and in some embodiments, wherein the point of attachment is a heteroatom contained in the heterocyclic ring. .

[0040] Thus, the term heterocycle includes aza-heterocyclyl, as used herein, which refers to a heterocyclyl containing at least one nitrogen, wherein the point of attachment of the heterocyclyl to the rest of the molecule is through a nitrogen atom in the heterocyclyl. Representative examples of aza-heterocyclyl include 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl. The term heterocycle also includes C-heterocyclyl, as used herein, which refers to a heterocyclyl containing at least one heteroatom, wherein the point of attachment of the heterocyclyl to the rest of the molecule is through a carbon atom in the heterocyclyl. Representative examples of carbo-heterocyclyl include 2-morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, and 2- or 3-pyrrolidinyl. The term heterocycle also includes heterocyclylalkyl groups, as described above, which refer to the formula—R c -heterocyclic group, wherein R c The term heterocycle also includes heterocyclylalkoxy, which, as used herein, refers to a heterocyclic group of the formula -OR c - a group bonded to the oxygen atom of a heterocyclic group, wherein R c It is an alkylene chain.

[0041] As used herein, the term "aryl" is used alone or as part of a larger moiety (e.g., "aralkyl", where the terminal carbon atom on the alkyl group is the point of attachment, e.g., benzyl), "aralkyloxy", where the oxygen atom is the point of attachment, or "aryloxyalkyl", where the point of attachment is on the aryl group, refers to a group including a monocyclic, bicyclic or tricyclic carbon ring system, including fused rings, where at least one ring in the system is aromatic. In some embodiments, the aralkyloxy is a phenoxy group. The term "aryl" can be used interchangeably with the term "aryl ring". In one embodiment, the aryl group includes a group having 6-18 carbon atoms. In another embodiment, the aryl group includes a group having 6-10 carbon atoms. Examples of aryl groups include phenyl, naphthyl, anthracenyl, biphenyl, phenanthrenyl, tetraphenyl, 1,2,3,4-tetrahydronaphthyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, naphthyridinyl, etc., which can be substituted or independently substituted by one or more substituents described herein. A special aryl group is phenyl. In some embodiments, aryl groups include an aryl ring fused to one or more (eg, 1, 2, or 3) different cyclic groups (eg, carbocyclic or heterocyclic), wherein the radical or point of attachment is on the aryl ring.

[0042] Thus, the term aryl includes aralkyl groups (eg, benzyl), which, as described above, refer to groups of the formula: c -aryl radical, wherein R c is an alkylene chain, such as methylene or ethylene. In some embodiments, the aralkyl group is an optionally substituted benzyl group. The term aryl also includes aralkyloxy groups. Aralkyloxy as used herein refers to a alkylene group of the formula --O--R c -- a group bonded to the oxygen atom of an aromatic group, wherein R c is an alkylene chain, such as methylene or ethylene.

[0043] As used herein, the term "heteroaryl", used alone or as part of a larger moiety (e.g., "heteroarylalkyl" (also known as "heteroaralkyl") or "heteroarylalkoxy" (also known as "heteroaralkoxy"), refers to a monocyclic, bicyclic or tricyclic ring system having 5 to 14 ring atoms, wherein at least one ring is aromatic and contains at least one heteroatom. In one embodiment, heteroaryl includes 4-6 membered monocyclic aromatic groups, wherein one or more ring atoms are independently optionally substituted nitrogen, sulfur or oxygen. In another embodiment, heteroaryl includes 5-6 membered monocyclic aromatic groups, wherein one or more ring atoms are nitrogen, Representative examples of heteroaryl groups include thienyl, furanyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidinyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, tetrazolyl[1,5-b]pyridazinyl, purinyl, deazapurinyl, benzoxazolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, indolyl, 1,3-thiazol-2-yl, 1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3, The term "heteroaryl" also includes groups in which the heteroaryl group is fused to one or more cyclic (e.g., carbocyclyl or heterocyclyl) rings, wherein the radical or point of attachment is on the heteroaryl ring. Non-limiting examples include indolyl, indolizinyl, isoindolyl, benzothiophenyl, benzothiophenyl, methylenedioxyphenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzodioxazolyl, benzothiazolyl, Quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolyl, carbazolyl, acridinyl, benzoxazinyl, phenothiazinyl, benzoxazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl and pyrido [2,3-b] -1,4-oxazin-3 (4H) -one. The heteroaryl group can be monocyclic, bicyclic or tricyclic. In some embodiments, the heteroaryl group includes a heteroaryl ring fused to one or more (e.g., 1, 2 or 3) different cyclic groups (e.g., carbocyclic or heterocyclic), wherein the group or point of attachment is on the heteroaryl ring, and in some embodiments, wherein the point of attachment is a heteroatom contained in the heterocyclic ring.

[0044] The term heteroaryl also includes N-heteroaryl, as used herein, which refers to a heteroaryl group as defined above containing at least one nitrogen atom, wherein the point of attachment of the N-heteroaryl group to the rest of the molecule is through the nitrogen atom in the heteroaryl group. The term heteroaryl also includes C-heteroaryl groups, as used herein, which refer to heteroaryl groups as defined above, and wherein the point of attachment of the heteroaryl group to the rest of the molecule is through a carbon atom in the heteroaryl group. The term heteroaryl also includes heteroarylalkyl groups, as disclosed above, which refer to groups of the formula -Rc-heteroaryl, wherein R c is an alkylene chain as defined above. The term heteroaryl also includes heteroaralkoxy (or heteroarylalkoxy) groups, which, as used herein, refer to groups bonded via an oxygen atom of the formula -O-Rc-heteroaryl, wherein R c is an alkylene group as defined above.

[0045] Unless otherwise indicated, and to the extent that no particular group is further defined, any group described herein may be substituted or unsubstituted. As used herein, the term "substituted" refers broadly to all permissible substituents, with the implicit condition that such substitution is determined according to the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, i.e., a compound that does not spontaneously undergo transformation (such as rearrangement, cyclization, elimination, etc.). Representative substituents include halogens, hydroxyls, and any other organic groups containing any number of carbon atoms (e.g., 1-14 carbon atoms), and which may include one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, sulfur, and nitrogen), grouped in linear, branched, or cyclic structures.

[0046] To the extent that any particular group is not disclosed, representative examples of substituents may thus include alkyl, substituted alkyl (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), substituted alkoxy (e.g., C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C1), halogen substituted alkyl (e.g., CF3), alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkenyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), substituted alkynyl (e.g., C2-C6, C2-C5, C2-C4, C2-C3, C2), cyclo (e.g., C3-C 12 , C5-C6), substituted rings (e.g., C3-C 12 , C5-C6), carbocyclic (e.g., C3-C 12, C5-C6), substituted carbocyclic rings (e.g., C3-C 12 , C5-C6), heterocyclic (e.g., C3-C 12 , C5-C6), substituted heterocycle (e.g., C3-C 12 , C5-C6), aryl (e.g., benzyl and phenyl), substituted aryl (e.g., substituted benzyl or phenyl), heteroaryl (e.g., pyridyl or pyrimidinyl), substituted heteroaryl (e.g., substituted pyridyl or pyrimidinyl), aralkyl (e.g., benzyl), substituted aralkyl (e.g., substituted benzyl), halogen, hydroxy, aryloxy (e.g., C6-C 12 , C6), substituted aryloxy (e.g., C6-C 12 , C6), alkylthio (e.g., C1-C6), substituted alkylthio (e.g., C1-C6), arylthio (e.g., C6-C 12 , C6), substituted arylthio (e.g., C6-C 12 , C6), cyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, thio, substituted thio, sulfinyl, substituted sulfinyl, sulfonyl, substituted sulfonyl, sulfinylimide, substituted sulfinylimide, sulfonamide, substituted sulfonamides, urea, substituted ureas, carbamate, substituted carbamate, amino acid and peptide groups.

[0047] The substituent may be a "nitrogen protecting group" (also known as an amino protecting group). Nitrogen protecting groups include, but are not limited to, -OH, -OR aa 、-N(R bb )2, -C(=O)R aa 、-C(=O)N(R bb 2. -CO2R aa 、-SO2R aa 、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-SO2N(R bb 2. -SO2R bb 、-SO2OR bb 、-SOR aa 、-C(=S)N(R bb )2, -C(=O)SR bb 、-C(=S)SR bb , C 1-10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2-10 Alkenyl, C 2-10Alkynyl, C 3-10 Carbocyclic group, 3-14 membered heterocyclic group, C 6-4 aryl and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R cc group substituted, and wherein R aa , R bb and R cc As defined herein. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3rd edition, John Wiley & Sons, 1999.

[0048] Nitrogen protecting groups such as amide groups (e.g., -C(=O)R aa Representative examples of ) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrobenzylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxyamido)acetamide, 3-(p-hydroxyphenyl)propionamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 2-methyl-2-(o-phenylazophenoxy)propionamide, 4-chlorobutyramide, 3-methyl-3-nitrobenzamide, o-nitrocinnamamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0049] Nitrogen protecting groups such as carbamate groups (e.g., -C(=O)OR aa) include but are not limited to methyl carbamate, ethyl carbamate, 9-fluorenylmethylcarbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluorenylmethylcarbamate, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthenyl)]carbamate (DBD-Tmoc), 4-methoxyphenylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), 2-trimethylsilylethylcarbamate (Teoc), 2-phenylethylcarbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1 , 1-dimethyl-2-haloethylcarbamate, 1,1-dimethyl-2,2-dibromoethylcarbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethylcarbamate, 1-methyl-1-(4-biphenylyl)ethylcarbamate (Bpoc), 1-(3,5-di-tert-butylphenyl)-1-methylethylcarbamate (t-Bumeoc), 2-(2- and 4-pyridyl)ethylcarbamate (Pyoc), 2-(N,N-dicyclohexylcarbamyl)ethylcarbamate, tert-butylcarbamate (BOC or Boc), 1-adamantylcarbamate (Adoc), vinylcarbamate (Voc), allylcarbamate (Alloc) , 1-isopropylallylcarbamate (Ipaoc), cinnamylcarbamate (Coc), 4-nitrocinnamylcarbamate (Noc), 8-quinolylcarbamate, N-hydroxypiperidinylcarbamate, alkyldithiocarbamate, benzylcarbamate (Cbz), p-methoxybenzylcarbamate (Moz), p-nitrobenzylcarbamate, p-bromobenzylcarbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzylcarbamate, 4-methylsulfonylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methylsulfonylethylcarbamate, 2-(p-toluenesulfonyl)ethylcarbamate methyl carbamate, [2-0,3-dicyano]carbamate (Dmoc), 4-methylthienylcarbamate (Mtpc), 2,4-dimethylthienylcarbamate (Bmpc), 2-phosphorylcarbamate (Peoc), 2-triphenylphosphorylisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxyboryl)benzylcarbamate, 5-benzisoxazolylmethylcarbamate, 2-(trifluoromethyl)-6-chromylmethylcarbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, o-nitrobenzylcarbamate, 3,4-dimethoxy-6-nitrobenzylcarbamate, methyl phenyl(o-nitrophenyl)carbamate, tert-amylcarbamate, S-benzylthiocarbamate, p-cyanobenzylcarbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentylcarbamate, cyclopropylmethylcarbamate, p-decyloxybenzylcarbamate, 2,2-dimethoxyacylvinylcarbamate, o-(N,N-dimethylformamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylformamido)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furylmethylcarbamate, 2-iodoethylcarbamate ester, isobornyl carbamate, isobutyl carbamate, isonicotinate, p-(p-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-tert-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate and 2,4,6-trimethylbenzyl carbamate.

[0050] Nitrogen protecting groups such as sulfonamide groups (e.g., -S(=O)2R aa ) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mrt), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), 13-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide and benzenesulfonamide.

[0051] Other nitrogen protecting groups include, but are not limited to, phenothiazine-(10)-acyl derivatives, N'-toluenesulfonylaminoacyl derivatives, N'-phenylaminosulfonyl, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiosuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilazide adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, 5-substituted substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro- 9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-pyridylmethylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)2,4,6-trimethylphenyl]methyleneamine, N-(N',N-dimethylmethylene)amine, N',N-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylboronic acid derivatives, N-[phenyl(pentacylchromium- or tungsten)acyl]amine, N-copper chelates, N-zinc chelates, N-nitrosamines, N-nitrosamines, amine N-oxides, diphenylphosphinamide (Dpp), dimethylphosphothioamide (Mpt), diphenylphosphothioamide (Ppt), dialkylphosphoramidates, dibenzylphosphoramidates, diphenylphosphoramidates, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide and 3-nitropyridinesulfenamide (Npys).

[0052] The substituent may be an "oxygen protecting group" (also known as a hydroxy protecting group). Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3rd edition, John Wiley & Sons, 1999, which are incorporated herein by reference. Exemplary oxygen protecting groups include, but are not limited to, methyl, tert-butyloxycarbonyl (Boc or BOC), methoxymethyl (MOM), methylthiomethyl (MTM), tert-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), tert-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloro ethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl, tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxane-2-yl, tetrahydrofuranyl, tetrahydrothiophenyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methylbenzofuran-2 -yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylseleno)ethyl, tert-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-methylpyridyl, 4-Methylpyridyl, 3-methyl-2-methylpyridyl N-oxide, diphenylmethyl, p-, p-dinitrophenyl, 5-dibenzodiaryl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4′-bromobenzoyloxyphenyl)diphenylmethyl, 4,4′,4″-tri(4,5-dichlorobenzimidyl)methyl, 4,4′,4″-tri(levulinyloxyphenyl)methyl, 4,4′,4″-tri(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfofuran-2-yl, benzisothiazolyl, S-dioxin, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylsilyl (dimethylthexylsilyl), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-dimethyl Phenylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), tert-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinic acid ester), 4,4-(ethylenedithio)pentaenoate (levulinic acid dithioacetal), pivalate, adamantanoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylenebenzoate, 2,4,6-trimethylbenzoate (methanesulfonate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), Alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Trot), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonyl)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl thiobenzyl carbonate, 4-ethoxy-1-naphthyl ethyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentyl carbonate esters, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, N,N,N',N'-tetramethylphosphoric acid diamide alkyl ester, N-phenylcarbamic acid alkyl ester, borate, dimethylphosphonyl, 2,4-Dinitrobenzenesulfinate alkyl esters, sulfates, mesylates (mesylate), benzyl sulfonates and toluene sulfonates (Ts).

[0053] The term "leaving group" is given its common meaning in the field of synthetic organic chemistry and refers to an atom or group that can be substituted by a nucleophile. For example, see Smith, March Advanced Organic Chemistry 6th edition (501-502). Examples of suitable leaving groups include, but are not limited to, halogen (such as F, Cl, Br or I), alkoxycarbonyloxy, aryloxycarbonyloxy, alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (such as acetoxy), arylcarbonyloxy, aryloxy, methoxy, N, O-dimethylhydroxylamino, 9-(9-phenyl)xanthenyl and haloformate. Exemplary leaving groups include, but are not limited to, activated substituted hydroxyl (e.g., -OC(=O)SR aa 、-OC(=O)R aa 、-OCO2R aa 、-OC(=O)N(R bb )2, -OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-OC(=NR bb )N(R bb )2. -OS(=O)R aa 、-OSO2R aa 、-OP(R cc )2.-OP(R cc )3, -OP(=O)2R aa ,-OP(=O)(R aa )2.-OP(=O)(OR cc )2、-OP(=O)2N(R bb )2 and -OP(=O)(NR bb )2, where R aa , R bb and R ccAs defined herein). In some cases, the leaving group is a sulfonate, such as a tosylate (tosylate, -OTs), a methanesulfonate (mesylate, -OMs), a bromobenzenesulfonyloxy (bromobenzenesulfonate, -OBs), -OS(=O)2(CF2)3CF3 (nonafluorobutanesulfonate, -ONf, or a trifluoromethanesulfonate (trifluoromethanesulfonate, -OTf). In some cases, the leaving group is a bromobenzenesulfonate, such as bromobenzenesulfonyloxy. In some cases, the leaving group is a nitrobenzenesulfonate, such as 2-nitrobenzenesulfonyloxy. The leaving group can also be a phosphinate (e.g., formed in a Mitsunobu reaction) or an internal leaving group, such as an epoxide or a cyclic sulfate. Other non-limiting examples of leaving groups are water, ammonia, alcohols, ether moieties, thioether moieties, zinc halides, magnesium moieties, diazonium salts, and copper moieties.

[0054] The term "binding" when referring to the interaction between a targeting ligand and a targeting protein (in the present invention, CDK7 and its mutant forms (collectively referred to as "CDK7")) generally refers to an intermolecular interaction that may be preferential or substantially specific (also referred to herein as "selective"), because the binding of the targeting ligand to other protein entities present in the cell is not functionally significant. The bispecific compounds of the present invention can preferentially bind to and recruit CDK7 and its mutant forms for targeted degradation.

[0055] The term "binding" when referring to the interaction between a degron and an E3 ubiquitin ligase generally refers to an intermolecular interaction that may or may not exhibit a level of affinity that equals or exceeds the affinity between a targeting ligand and a target protein, but nonetheless wherein the affinity is sufficient to achieve targeted and selective degradation of the target protein by recruiting the ligase to the target protein.

[0056] In broad terms, bispecific compounds include a portion that binds to cyclin-dependent kinase 7 (CDK7) (referred to herein as a targeting ligand) and a second portion that binds to an E3 ubiquitin ligase (referred to as a "degradant"), which are linked together by a linker. The compound has a structure represented by formula (I):

[0057] or a pharmaceutically acceptable salt or stereoisomer thereof, wherein

[0058] R1 stands for -NR a R b 、-CHR a R b OR a , where R a and R beach independently represents hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, or an oxygen protecting group when attached to an oxygen atom, or R a and R b together with the atoms to which they are attached, form an optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl or optionally substituted heteroaryl ring;

[0059] R3 and R4 each independently represent hydrogen, halogen, optionally substituted C1-C6 alkyl or optionally substituted aryl, or R3 and R4 together with the atoms to which they are attached form an optionally substituted C3-C6 carbocyclyl ring;

[0060] R5 independently represents hydrogen, optionally substituted C1-C6 alkyl or a nitrogen protecting group;

[0061] L1 stands for -NR L1 -、-NR L1 C(=O)-, -C(=O)NR L1 -, -O- or -S-, where R L1 is hydrogen, optionally substituted C1-C6 alkyl or a nitrogen protecting group;

[0062] A represents an optionally substituted carbocyclic group, an optionally substituted heterocyclic group, an optionally substituted aryl group or an optionally substituted heteroaryl group;

[0063] L2 represents a bond or does not exist, -C(=O)-, -C(=O)NR L2 -、-NR L2 C(=O)-, -O- or -S-, wherein R L2 is hydrogen, optionally substituted C1-C6 alkyl or a nitrogen protecting group;

[0064] B is a bond or absent, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; and

[0065] R2 is absent, or any of the following structures:

[0066]

[0067]

[0068] in

[0069] Asterisk ( * ) represents the connection point with B, and the curved line represents The connection point;

[0070] L3 is a bond or absent or optionally substituted C 1-4 A hydrocarbon chain, wherein one or more carbon units of the hydrocarbon chain are independently replaced by -C(=O)-, -O-, -S-, -NR L3a 、-NR L3a C(=O)-, -C(=O)NR L3a -, -SC(=O)-, -C(=O)S-, -OC(=O)-, -C(=O)O-, -NR L3a C(=S)-、-C(=S)NR L3a -, trans-CR L3b =CR L3b -、cis-CR L3b =CR L3b -, -CS(=O)-, -C≡C-, -S(=O)-, -S(=O)O-, -OS(=O)-, -S(=O)NR L3a -、-NR L3a S(=O)-, -S(=O)2-, -S(=O)2O-, -OS(=O)2-, -S(=O)2NR L3a -or-NR L3a S(=O)2-substituted, wherein R L3a is hydrogen, substituted or unsubstituted C 1-6 alkyl or a nitrogen protecting group, and wherein R L3b Each occurrence of is independently hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R L3b The groups, together with the atoms to which they are attached, form an optionally substituted carbocyclic ring or an optionally substituted heterocyclic ring;

[0071] L4 is a bond or an optionally substituted, branched or unbranched C 1-6 hydrocarbon chain;

[0072] R EI , R E2 and R E3 each independently represents hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, -CN, -CH2OR EE 、-CH2N(R EE )2, -CH2SR EE 、-OR EE 、-N(R EE )2、-Si(R EE )3 and -SR EE , where R EEEach occurrence of is independently hydrogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R EE The groups, together with the atoms to which they are attached, form an optionally substituted heterocyclic ring; or R E1 and R E3 , or R E2 and R E3 , or R E1 and R E2 together with the atoms to which they are attached, form an optionally substituted carbocyclic ring or an optionally substituted heterocyclic ring;

[0073] R E4 is a leaving group;

[0074] R E6 is hydrogen, substituted or unsubstituted C 1-6 Alkyl or nitrogen protecting groups;

[0075] Each instance of Y is independently O, S or NR E7 , where R E7 is hydrogen, substituted or unsubstituted C 1-6 Alkyl or nitrogen protecting groups;

[0076] a is 1 or 2; and

[0077] Each instance of z is independently 0, 1, 2, 3, 4, 5 or 6, as valency permits.

[0078] With respect to the compounds of the present invention, the targeting ligand is represented by the following structure:

[0079]

[0080] The curve represents connection point.

[0081] In some embodiments, R1 is in

[0082] Each R 1’ YesR 1” are independently hydrogen, optionally substituted C1-C6 alkyl or a nitrogen protecting group,

[0083] R 1a is hydrogen, C1-C6 alkyl or optionally substituted aryl, and

[0084] R 2a is hydrogen, -OR 1N or -NR 1N R 2N , where R1N and R 2N Each is independently hydrogen, C1-C6 alkyl, or a nitrogen protecting group when attached to a nitrogen or an oxygen protecting group when attached to an oxygen atom.

[0085] In some embodiments, R 1” is hydrogen, Bn, BOC, Cbz, Fmoc, trifluoroacetyl, triphenylmethyl, acetyl or Ts.

[0086] In some embodiments, R1 is In some embodiments, R1 is

[0087] In some embodiments, R 1a is hydrogen, methyl, ethyl, propyl or phenyl.

[0088] In certain embodiments, R 2a is hydrogen. In some embodiments, R 2a Yes-OR IN , where R 1N is hydrogen, C1-C6 alkyl or an oxygen protecting group. 2a In certain embodiments, R 2a Yes - NR 1N R 2N , where R IN and R 2N Each is independently hydrogen, C1-C6 alkyl or a nitrogen protecting group. 1N and R 2N are hydrogen, methyl, ethyl, propyl or a nitrogen protecting group. 1N and R 2N At least one of is hydrogen, methyl, ethyl, propyl or a nitrogen protecting group. IN is a methyl group, and R 2N is hydrogen. In some embodiments, R 1N is ethyl, and R 2N is hydrogen. In some embodiments, R 1N is propyl, and R 2N is hydrogen. In some embodiments, R 1N is a nitrogen protecting group, and R 2N is hydrogen. In some embodiments, R 1N is methyl, R 2N is a nitrogen protecting group. 1N is ethyl, R 2N is a nitrogen protecting group. 1N is propyl, R2N is a nitrogen protecting group.

[0089] In some embodiments, R1 is

[0090] In some embodiments, R2 is a bond,

[0091] In some embodiments, R3 and R4 are independently methyl, isopropyl or phenyl, or R3 and R4 together with the atoms to which they are attached form an optionally substituted C3-C6 carbocyclyl ring. In some embodiments, R3 and R4 are both methyl.

[0092] In some embodiments, R5 is hydrogen or methyl.

[0093] In some embodiments, A is a 6-membered carbocyclyl or a 6-membered heterocyclyl.

[0094] In some embodiments, B is a 6-membered carbocyclyl or a 6-membered heterocyclyl.

[0095] In some embodiments, L1 is L1 is NH or -NHC(O)-.

[0096] In some embodiments, L2 is a bond, NH, or -NHC(O)-.

[0097] In some embodiments, wherein L1 is NH or -NHC(O)-, R3 and R4 are methyl, and R5 is H, the compound of formula (I) has a structure represented by any one of formulas (I-1a) and (I-1b):

[0098]

[0099] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0100] In some embodiments, wherein A is a 6-membered carbocyclyl, R3 and R4 are methyl, R5 is H, the compound of formula (I) has a structure represented by any one of formulas (I-2a) to (I-2f):

[0101]

[0102]

[0103] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0104] In some embodiments, wherein L1 is NH or -NHC(O)-, R1 is R3 and R4 are methyl groups, and R5 is H, and the compound of formula (I) has a structure represented by any one of formulas (I-3a) to (I-3d):

[0105]

[0106]

[0107] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0108] In some embodiments, wherein A is a 6-membered carbocyclyl, R1 is R3 and R4 are methyl groups, and R5 is H, and the compound of formula (I) has a structure represented by any one of formulas (I-4a) to (I-41):

[0109]

[0110]

[0111]

[0112]

[0113] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0114] In some embodiments, wherein L1 is NH or -NHC(O)-, R1 is R3 and R4 are methyl groups, and R5 is H, and the compound of formula (I) has a structure represented by any one of formulas (I-5a) and (I-5b):

[0115]

[0116]

[0117] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0118] In some embodiments, wherein A is a 6-membered carbocyclyl, R1 is R3 and R4 are methyl groups, and R5 is H, and the compound of formula (I) has a structure represented by any one of formulas (I-6a) to (I-61):

[0119]

[0120]

[0121]

[0122]

[0123] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0124] In some embodiments, the compound of formula (I) has a structure represented by any one of formulas (I-7) to (I-57):

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0141] In some embodiments, wherein L1 is -NHC(O)-, A is a 6-membered carbocyclyl, each of B, L2, and R2 is a bond (or absent), R1 is And R3 and R4 are both methyl groups, the compound of formula (I) has a structure represented by formula (I-58):

[0142]

[0143] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0144] Connectors

[0145] The linker ("L") provides a covalent link between the targeting ligand and the degron. The structure of the linker may not be important, as long as it does not substantially interfere with the activity of the targeting ligand or the degron. In some embodiments, the linker comprises an alkylene chain (e.g., having 2-20 alkylene units). In other embodiments, the linker may include an alkylene chain or a divalent alkylene chain, any of which may be replaced by -O-, -S-, -N(R')-, -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)-, -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(Me)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-C 12 At least one of a carbocyclylene, a 3- to 12-membered heterocyclylene, a 5- to 12-membered heteroarylene, or any combination thereof is interrupted and / or terminated (at one or both ends), wherein R' is H or a C1-C6 alkyl group, wherein the interrupting group and one or both terminating groups may be the same or different.

[0146] In some embodiments, the linker may include a C1-C 12 An alkylene chain, wherein the nitrogen is also bound to the degron.

[0147] In certain embodiments, the linker comprises a Interruption or termination of an alkylene chain.

[0148] "Carbocyclylene" refers to an optionally substituted divalent carbocyclic group.

[0149] "Heterocyclylene" refers to a divalent heterocyclic group which may be optionally substituted.

[0150] "Heteroarylene" refers to a divalent heteroaryl group which may be optionally substituted.

[0151] Representative examples of linkers suitable for use in the present invention include the following:

[0152] wherein n is an integer from 1 to 12 (inclusive), for example, 1-12, 1-11, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, 9-10 and 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, examples of which include:

[0153]

[0154] Alkylene chains terminated in various functional groups (as described above) are exemplified by the following:

[0155]

[0156] Alkylene chains interrupted by various functional groups (as described above) are exemplified by the following:

[0157]

[0158] An alkylene chain interrupted or terminated with a heterocyclylene group, e.g.

[0159] Wherein m and n are independently integers of 0-10, examples of which include:

[0160]

[0161]

[0162] Examples of alkylene chains interrupted by amide, heterocyclylene and / or aryl groups include:

[0163]

[0164] Examples of alkylene chains interrupted by heterocyclylene and aryl groups and heteroatoms include:

[0165]

[0166] and

[0167] An alkylene chain interrupted by heteroatoms such as nitrogen, oxygen or boron, e.g.

[0168] wherein each n is independently an integer from 1 to 10, for example, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, 9-10 and 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, and R is hydrogen or C1 to C4 alkyl, examples of which are

[0169]

[0170] In some embodiments, the linker may include a polyethylene glycol chain, which may terminate (at either or both ends) with -S-, -N(R')-, -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)-, -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(Me)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')-, C 3-12 At least one of a carbocyclylene, a 3- to 12-membered heterocyclylene, a 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or a C1-C6 alkyl, wherein one or both of the terminating groups may be the same or different.

[0171] In some embodiments, the linker comprises a PEGylated oligomer having 2-8 PEG units and terminating in of polyethylene glycol chains.

[0172] Examples of linkers comprising polyethylene glycol chains include:

[0173] Where n is an integer from 2 to 10, examples of which include:

[0174]

[0175] In some embodiments, the polyethylene glycol linker can terminate in a functional group, examples of which are as follows:

[0176]

[0177]

[0178] In some embodiments, the compound of Formula (I) includes a linker represented by structure (L10):

[0179]

[0180] Wherein Q is CH2 or O;

[0181] Y is CH2, CH2CH2, or absent, provided that when X is O, Y is CH2CH2; and n is an integer from 0 to 6.

[0182] In some embodiments, the linker is represented by any of the structures L11-L23:

[0183]

[0184]

[0185] In some embodiments, the bispecific compound of Formula (I) has a structure represented by any one of Formulas (I-59) to (I-71):

[0186]

[0187]

[0188]

[0189] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0190] Degron

[0191] A degron ("D") is a functional moiety that binds an E3 ubiquitin ligase. In some embodiments, a degron binds cereblon (CRBN).

[0192] Representative examples of degrons that bind cereblon and may be suitable for use in the present invention are described in U.S. Patent Application Publication No. 2018 / 0015085 (e.g., indolones such as isoindolinone and isoindolin-1,3-dione encompassed by Formulas IA and IA', and bridged cycloalkyl compounds encompassed by Formulas 1B and 1B').

[0193] In some embodiments, the compound of Formula (I) includes a cereblon-binding degron represented by any one of Structures (D1-a) to (D1-h):

[0194]

[0195] In some embodiments, the compound of Formula (I) has a structure represented by any one of Formulas (I-72a) to (I-72h):

[0196]

[0197]

[0198] or a pharmaceutically acceptable salt or stereoisomer thereof.

[0199] In some embodiments, the degron binds to the Von Hippel-Lindau (VHL) tumor suppressor. Representative examples of degrons that bind to VHL are as follows:

[0200]

[0201]

[0202] , wherein Y' is a bond, N, O or C;

[0203] wherein Z is a C5-C6 carbocyclic or C5-C6 heterocyclic group; and In some embodiments, Z is

[0204] U.S. Patent Application Publication No. 2017 / 0121321A1 discloses other degrons that bind VHL and may be suitable for use as degrons in the present invention.

[0205] In some embodiments, the compound of Formula (I) has a structure represented by any one of Formulas (I-73a) to (I-73e):

[0206]

[0207] wherein R represents H, methyl, ethyl, isopropyl or CF3; Y' is a bond, N, O or C; and Z is a C5-C6 carbocyclic or heterocyclic group; or a pharmaceutically acceptable salt or stereoisomer thereof.

[0208] In some embodiments, the compound of Formula (I) is represented by any one of Structures (1) to (26):

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217] or pharmaceutically acceptable salts and stereoisomers thereof.

[0218] The bispecific compound of formula (I) may be in the form of a free acid or free base or a pharmaceutically acceptable salt. As used herein, in the context of a salt, the term "pharmaceutically acceptable" refers to a salt of a compound that does not eliminate the biological activity or properties of the compound and is relatively non-toxic, i.e., the compound in salt form can be administered to a subject without causing undesirable biological effects (such as dizziness or stomach discomfort) or interacting in a harmful manner with any other component of the composition comprising it. The term "pharmaceutically acceptable salt" refers to a product obtained by reacting a compound of the invention with a suitable acid or base. Examples of pharmaceutically acceptable salts of the compounds of the invention include those derived from suitable inorganic bases such as lithium, sodium, potassium, calcium, magnesium, iron, copper, aluminum, zinc and manganese salts. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, dextran salt, sucrose salt, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, 4-methylbenzenesulfonate or p-toluenesulfonate, etc. Certain compounds of the present invention can form pharmaceutically acceptable salts with various organic bases such as lysine, arginine, guanidine, diethanolamine or metformin.

[0219] The bispecific compounds of formula (I) may have at least one chiral center. Therefore, they may be in the form of stereoisomers. As used herein, the term "stereoisomer" includes all isomers of a single compound, the difference between which isomers is that their atoms are oriented differently in space. The term stereoisomer includes mirror image isomers (enantiomers including (R-) or (S-) configurations of compounds), mixtures of mirror image isomers of compounds (physical mixtures and racemates or racemic mixtures of enantiomers), geometric (cis / trans or E / Z, R / S) isomers of compounds, and isomers (diastereomers) of compounds that have more than one chiral center but are not mirror images of each other. The chiral center of the compound may undergo epimerization in vivo; therefore, for these compounds, compounds administered in (R-) form are considered equivalent to compounds administered in (S-) form. Thus, the compounds of the present invention may be prepared and used as an individual isomer, substantially free of other isomers, or as a mixture of various isomers, for example, a racemic mixture of stereoisomers.

[0220] In some embodiments, the bispecific compound of Formula (I) is an isotopic derivative in that it has a desired isotopic substitution of at least one atom that is greater than the natural abundance of the isotope, i.e., is enriched. In one embodiment, the compound includes deuterium or a plurality of deuterium atoms. 2 H)) substitution may offer certain therapeutic advantages due to greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be advantageous in some circumstances.

[0221] In addition to isotopic derivatives, the term "bispecific compound of Formula (I)" includes the use of N-oxides, crystalline forms (also known as polymorphs), active metabolites of compounds having the same type of activity, tautomers, and unsolvated and solvated forms made with pharmaceutically acceptable solvents (e.g., water, ethanol, etc.). The solvated forms of the conjugates given herein are also considered to be disclosed herein.

[0222] Synthesis method

[0223] In some embodiments, the present invention relates to a method for preparing a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof. In a broad sense, the compound of the present invention or a pharmaceutically acceptable salt or stereoisomer thereof can be prepared by any known method suitable for preparing chemically related compounds. Representative synthetic schemes are described in various working examples, which illustrate non-limiting methods for preparing the compounds of the present invention.

[0224] Pharmaceutical composition

[0225] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. As known in the art, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle suitable for administering the compounds of the present invention to mammals. Suitable carriers may include, for example, liquids (aqueous and non-aqueous, and combinations thereof), solids, encapsulating materials, gases and combinations thereof (e.g., semisolids), and gases, the function of which is to transport or deliver a compound from one organ or part of the body to another organ or another part of the body. The carrier is "acceptable", meaning that it is physiologically inert and compatible with the other ingredients of the preparation, and is harmless to the subject or patient. Depending on the type of preparation, the composition may include one or more pharmaceutically acceptable excipients.

[0226] In broad terms, the bispecific compounds of formula (I) and pharmaceutically acceptable salts and stereoisomers thereof can be formulated into a composition of a given type according to conventional pharmaceutical practices such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping and compression processes (see, e.g., Remington: The Science and Practice of Pharmacy (20th ed.), ed. A. R. Gennaro, Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York). The type of formulation depends on the mode of administration, which may include enteral (e.g., oral, buccal, sublingual, and rectal), parenteral (e.g., subcutaneous (sc), intravenous (iv), intramuscular (im), and intrasternal injection, or infusion techniques, intraocular, intraarterial, intramedullary, intrathecal, intraventricular, transdermal, intradermal, intravaginal, intraperitoneal, mucosal, nasal, intratracheal instillation, bronchial instillation, and inhalation), and topical (e.g., transdermal). In general, the most appropriate route of administration depends on a variety of factors, including, for example, the nature of the medicament (e.g., its stability in the gastrointestinal environment) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration). For example, parenteral (e.g., intravenous) administration is also advantageous because the compound can be administered relatively quickly, for example, in the case of single-dose treatment and / or acute conditions.

[0227] In some embodiments, the compounds are formulated for oral or intravenous administration (eg, systemic intravenous injection).

[0228] Thus, the bispecific compounds of the invention can be formulated into solid compositions (e.g., powders, tablets, dispersible particles, capsules, cachets, and suppositories), liquid compositions (e.g., solutions in which the compound is dissolved, suspensions in which solid particles of the compound are dispersed, emulsions, and solutions containing liposomes, micelles, or nanoparticles, syrups, and elixirs); semisolid compositions (e.g., gels, suspensions, and creams); and gases (e.g., propellants for aerosol compositions). The compounds can also be formulated for rapid, intermediate, or extended release.

[0229] Oral solid dosage forms include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with a carrier (such as sodium citrate or dicalcium phosphate) and additional carriers or excipients such as a) fillers or extenders (such as starch, lactose, sucrose, glucose, mannitol and silicic acid), b) binders (such as methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia), c) humectants (such as glycerol), d) disintegrants (such as cross-linked polymers (e.g., Cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium starch glycolate, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate), e) solution retardants (such as paraffin), f) absorption promoters (such as quaternary ammonium compounds), g) wetting agents (such as cetyl alcohol and glycerol monostearate), h) absorbents (such as kaolin and bentonite), and i) lubricants (such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate) and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also include a buffer. Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or lactose and high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells (such as enteric coatings and other coatings). They may further contain opacifiers.

[0230] In some embodiments, the bispecific compounds of the invention can be formulated in hard or soft gelatin capsules. Representative excipients that can be used include pregelatinized starch, magnesium stearate, mannitol, sodium stearyl fumarate, anhydrous lactose, microcrystalline cellulose, and cross-linked sodium carboxymethyl cellulose. The gelatin shell can include gelatin, titanium dioxide, iron oxide, and a colorant.

[0231] Oral liquid dosage forms include solutions, suspensions, emulsions, microemulsions, syrups and elixirs. In addition to the compound, the liquid dosage form can include aqueous or non-aqueous carriers commonly used in the art (depending on the solubility of the compound), such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof. Oral compositions may also include excipients, such as wetting agents, suspending agents, coloring agents, sweeteners, flavoring agents and flavoring agents.

[0232] Injectable preparations may include sterile aqueous solutions or oily suspensions. They may be prepared using suitable dispersants or wetting agents and suspending agents according to standard techniques. Sterile injectable preparations may also be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable carriers and solvents include water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as solvents or suspension media. For this purpose, any mild fixed oil may be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) are used to prepare injections. Injectable preparations may be sterilized, for example, by filtering through a bacterial retention filter, or by incorporating a sterilizing agent in the form of a sterile solid composition, which may be dissolved or dispersed in sterile water or other sterile injection media before use. The effect of the compound may be prolonged by slowing its absorption, which may be achieved by using a poorly water-soluble liquid suspension or a crystalline or amorphous material. The extended absorption of the compound from parenteral administration preparations may also be achieved by suspending the compound in an oily vehicle.

[0233] In certain embodiments, the bispecific compound of formula (I) can be applied in a local rather than systemic manner, for example, by injecting the conjugate directly into an organ, usually in the form of a depot preparation or a sustained release preparation. In a specific embodiment, the long-acting preparation is applied by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. The injectable depot form is prepared by forming a microcapsule matrix of the compound in a biodegradable polymer, such as polylactide-polyglycolide, poly (orthoester) and poly (anhydride). The release rate of the compound can be controlled by changing the ratio of the compound to the polymer and the properties of the specific polymer used. The injectable depot preparation is also prepared by encapsulating the compound in a liposome or microemulsion compatible with body tissues. In addition, in other embodiments, the compound is delivered in a targeted drug delivery system, for example, in a liposome coated with an organ-specific antibody. In such an embodiment, the liposome targets the organ and is selectively absorbed by the organ.

[0234] The bispecific compounds of the invention may be formulated for buccal or sublingual administration, examples of which include tablets, lozenges and gels.

[0235] The bispecific compound can be formulated for inhalation administration. Various forms suitable for inhalation administration include aerosols, mists or powders. The pharmaceutical composition can be delivered in an aerosol form from a pressurized package or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In some embodiments, the dosage unit of the pressurized aerosol can be determined by providing a valve to deliver a metered amount. In some embodiments, capsules and cartridges containing gelatin, such as for use in an inhaler or insufflator, can be formulated into a powder mixture containing the compound and a suitable powder base (such as lactose or starch).

[0236] The bispecific compounds of formula (I) may be formulated for topical administration, which as used herein refers to intradermal administration by applying the formulation to the epidermis. These types of compositions are typically in the form of ointments, pastes, creams, lotions, gels, solutions and sprays.

[0237] Representative examples of carriers for preparing compositions for topical application include solvents (e.g., alcohols, polyols, water), creams, lotions, ointments, oils, plasters, liposomes, powders, emulsions, microemulsions, and buffered solutions (e.g., hypotonic or buffered saline). Creams, for example, can be prepared using saturated or unsaturated fatty acids (e.g., stearic acid, palmitic acid, oleic acid, palmitoleic acid, cetyl alcohol, or oleyl alcohol). Creams can also contain nonionic surfactants, such as polyoxyethylene (40) stearate.

[0238] In some embodiments, topical preparations may also include excipients, one example of which is a penetration enhancer. These agents can transport pharmacologically active compounds through the stratum corneum and into the epidermis or dermis, preferably with little or no systemic absorption. The effectiveness of various compounds in improving the rate at which drugs penetrate the skin has been evaluated. See, for example, Percutaneous Penetration Enhancers, Maibach HI and Smith HE (eds.), CRC Press, Inc., Boca Raton, Fla. (1995), which investigated the use and testing of various skin penetration enhancers, and Buyuktimkin et al., Chemical Means of Transdermal Drug Permeation Enhancement in Transdermal and Topical Drug Delivery Systems, Gosh TK, Pfister WR, Yum S.I. (Eds.), Interpharm Press Inc., Buffalo Grove, Ill. (1997). Representative examples of penetration enhancers include triglycerides (e.g., soybean oil), aloe compositions (e.g., aloe vera gel), ethanol, isopropyl alcohol, octadecylphenyl polyethylene glycol, oleic acid, polyethylene glycol 400, propylene glycol, N-decyl methyl sulfoxide, fatty acid esters (e.g., isopropyl myristate, methyl laurate, glycerol monooleate and propylene glycol monooleate), and N-methylpyrrolidone.

[0239] Representative examples of other excipients that may be included in topical preparations and other types of preparations (to the extent that they are compatible) include preservatives, antioxidants, humectants, emollients, buffers, solubilizers, skin protectants and surfactants. Suitable preservatives include alcohols, quaternary amines, organic acids, parabens and phenols. Suitable antioxidants include ascorbic acid and its esters, sodium bisulfite, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol and chelating agents such as ethylenediaminetetraacetic acid and citric acid. Suitable humectants include glycerol, sorbitol, polyethylene glycol, urea and propylene glycol. Suitable buffers include citric acid, hydrochloric acid and lactic acid buffers. Suitable solubilizers include quaternary ammonium chlorides, cyclodextrins, benzyl benzoate, lecithin and polysorbate. Suitable skin protectants include vitamin E oil, allantoin, dimethicone, glycerol, vaseline and zinc oxide.

[0240] Transdermal preparations generally use transdermal application devices and transdermal application patches, wherein the compound is formulated into a lipophilic emulsion or a buffered aqueous solution, dissolved and / or dispersed in a polymer or adhesive. The patch can be configured to deliver a medicament continuously, pulsatilely or on demand. The transdermal delivery of a compound can be accomplished by an iontophoresis patch. A transdermal patch can provide controlled delivery of a compound, wherein the absorption rate is slowed down by using a rate-controlled membrane or by trapping the compound in a polymer matrix or a gel. Absorption enhancers can be used to increase absorption, and examples thereof include absorbable pharmaceutically acceptable solvents that help pass through the skin.

[0241] Ophthalmic preparations include eye drops.

[0242] Formulations for rectal administration include enemas, rectal gels, rectal foams, rectal aerosols and retention enemas, which may contain conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinyl pyrrolidone, polyethylene glycol, etc. Compositions for rectal or vaginal administration may also be formulated as suppositories, which can be prepared by mixing the compound with suitable non-irritating carriers and excipients such as cocoa butter, mixtures of fatty acid glycerides, polyethylene glycol, suppository waxes, and combinations thereof, all of which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the compound.

[0243] dose

[0244] As used herein, the term "therapeutically effective amount" refers to an amount of a bispecific compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof that is effective in producing a desired therapeutic response in a particular patient suffering from a disease or condition characterized by or mediated by abnormal CDK7 activity. The term "therapeutically effective amount" therefore includes an amount of a compound of the invention or a pharmaceutically acceptable salt or stereoisomer thereof that, when administered, induces a positive change in the disease or condition to be treated (e.g., to selectively inhibit / degrade FGFR2), or is sufficient to prevent the development or progression of a disease or condition, or to alleviate to some extent one or more symptoms of the disease or condition being treated in a subject, or to simply kill or inhibit the growth of diseased (e.g., neuroblastoma) cells, or to reduce the amount of CDK7 in diseased cells.

[0245] The total daily dose of the bispecific compound and its use can be determined according to standard medical practice, for example, by the attending physician using sound medical judgment. The specific therapeutically effective dose for any particular subject may depend on a variety of factors, including the disease or condition being treated and its severity (e.g., its current state); the subject's age, weight, general health, sex, and diet; the administration time, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination or concurrently with the bispecific compound; and factors well known in the medical field (see, for example, Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 10th Edition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press, 155-173, 2001).

[0246] The bispecific compounds of formula (I) and pharmaceutically acceptable salts and stereoisomers thereof can be effective over a wide dosage range. In some embodiments, the total daily dose (e.g., for adults) can be in the range of about 0.001 to about 1600 mg, 0.01 to about 1600 mg, 0.01 to about 500 mg, about 0.01 to about 100 mg, about 0.5 to about 100 mg, 1 to about 100-400 mg per day, about 1 to about 50 mg per day, about 5 to about 40 mg per day, and in other embodiments, about 10 to about 30 mg per day. Depending on the number of times the compound is administered per day, a single dose can be formulated to contain the desired dose. For example, a capsule can be formulated with about 1 to about 200 mg of a bispecific compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof (e.g., 1, 2, 2.5, 3, 4, 5, 10, 15, 20, 25, 50, 100, 150, and 200 mg). In some embodiments, depending on how many times per day the compound is administered, a single dosage can be formulated to contain the desired dosage.

[0247] How to use

[0248] In some aspects, the invention relates to methods for treating diseases or conditions involving abnormal (e.g., dysfunctional or dysregulated) CDK7 activity, which requires administering to a subject in need thereof a therapeutically effective amount of a bispecific compound of Formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0249] It can be said that these diseases or disorders are characterized by abnormal (e.g., loss of function or disorder) FGFR2 activity or mediated by abnormal CDK7 activity (e.g., relative to a non-pathological state, the level of the protein is increased or functionally abnormal). "Disease" is generally considered to be a health condition of a subject, in which the subject cannot maintain homeostasis, and in which if the disease is not improved, the health of the subject continues to deteriorate. In contrast, a subject's "disorder" refers to a health condition in which the subject is able to maintain homeostasis, but the health condition of the subject is not as good as the health condition when there is no disease. If not treated, the disease will not necessarily lead to a further decline in the health condition of the animal.

[0250] The term "subject" (or "patient") as used herein includes all members of the animal kingdom susceptible to or suffering from the disease or condition. In some embodiments, the subject is a mammal, e.g., a human or non-human mammal. These methods are also applicable to companion animals, such as dogs and cats, and livestock, such as cattle, horses, sheep, goats, pigs and other domestic and wild animals. A subject who "needs" treatment according to the present invention may "suffer from or be suspected of suffering from" a specific disease or condition, and the subject may have been diagnosed or otherwise present with a sufficient number of risk factors or a sufficient number or combination of signs or symptoms so that a medical professional can diagnose or suspect that the subject suffers from the disease or condition. Therefore, subjects suffering from and suspected of suffering from a specific disease or condition are not necessarily two different groups.

[0251] Exemplary types of non-cancerous (e.g., cell proliferative) diseases or conditions that can be treated with the compounds of the invention include inflammatory diseases and conditions, autoimmune diseases, neurodegenerative diseases, cardiac diseases, viral diseases, chronic and acute kidney disease or injury, metabolic diseases, and allergic and genetic diseases.

[0252] Representative examples of specific non-cancerous diseases and disorders include lymphoproliferative conditions, autoimmune diseases, cholecystitis, acromegaly, rheumatoid spondylitis, osteoarthritis, gout, sepsis, septic shock, dacryoadenitis, protein-associated periodic syndrome (CAPS), endotoxin shock, endometritis, gram-negative sepsis, keratoconjunctivitis sicca, toxic shock syndrome, asthma, adult respiratory distress syndrome, chronic obstructive pulmonary disease, chronic pulmonary Inflammation, chronic transplant rejection, hidradenitis suppurativa, inflammatory bowel disease, Crohn's disease, Behcet's syndrome, glomerulonephritis, multiple sclerosis, juvenile diabetes, thyroiditis, Addison's disease, appendicitis, granulomatous orchitis, eczema, pancreatic fibrosis, hepatitis, liver fibrosis, CD14-mediated sepsis, non-CD14-mediated sepsis, acute and chronic kidney disease, irritable bowel syndrome, fever, restenosis, cervicitis, stroke and ischemic injury, nerve damage, acute allergic rhinitis, allergic conjunctivitis, chronic heart failure, congestive heart failure, acute coronary syndrome, cachexia, malaria, acute and chronic heart failure, congestive heart failure, acute coronary syndrome, cachexia, malaria, leprosy, leishmaniasis, Lyme disease, Rett's syndrome, acute synovitis, muscle degeneration, bursitis, tendinitis, tenosynovitis, herniated, ruptured or prolapsed disc syndrome, osteosclerosis, sinusitis, thrombosis, silicosis, pulmonary myopathy, bone resorption diseases (such as osteoporosis), fibromyalgia, AIDS and other viral diseases (such as herpes zoster, herpes simplex I or II, influenza virus and cytomegalovirus), type I and II diabetes, obesity, insulin resistance and diabetic retinopathy, 22q11.2 deletion syndrome, Angelman syndrome, Canavan disease, Charcot-Marie-Tooth disease, color blindness, Cri du chat, Down syndrome, cystic fibrosis, Duchenne muscular dystrophy, hemophilia, Klinefleter syndrome, neurofibromatosis, phenylketonuria, Prader-Willi syndrome, sickle cell disease, Tay-Sachs disease, Turner syndrome, urea cycle disorders, thalassemia, otitis media, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, pneumonia, uveitis, polymyositis, proctitis, interstitial pulmonary fibrosis, dermatomyositis, atherosclerosis, arteriosclerosis, amyotrophic lateral sclerosis, vascular dementia, varicose veins, vaginitis, depression, and sudden infant death syndrome.

[0253] Examples of autoimmune diseases include autoimmune blood diseases (e.g., hemolytic anemia, aplastic anemia, anhidrotic ectodermal dysplasia, pure red cell anemia, and idiopathic thrombocytopenia), alopecia areata, rheumatoid arthritis, scleroderma, systemic lupus erythematosus, autoimmune uveoretinitis, autoimmune vasculitis, lichen planus, bullous pemphigus, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, myasthenia gravis, immunoglobulin A nephropathy, Hashimoto's disease, Sjögren's syndrome, vitiligo, Wegener's granulomatosis, autoimmune oophoritis, sarcoidosis, rheumatic carditis, ankylosing spondylitis, Graves' disease, autoimmune thrombocytopenic purpura, psoriasis, psoriatic arthritis, dermatitis herpetiformis, ulcerative colitis, and celiac disease.

[0254] In other embodiments, the method is directed to treating a subject with cancer. In general, the bispecific compounds of the invention are effective in treating carcinomas (solid tumors, including primary and metastatic tumors), sarcomas, melanomas, and hematologic cancers (cancers that affect the blood, including lymphocytes, bone marrow, and / or lymph nodes), such as leukemias, lymphomas, and multiple myeloma. Both adult tumors / cancers and pediatric tumors / cancers are included. Cancers can be vascularized, or not substantially vascularized, or non-vascularized tumors.

[0255] Representative examples of cancer include adrenocortical carcinoma, AIDS-related cancers (e.g., Kaposi's and AIDS-related lymphomas), appendix cancer, childhood cancers (e.g., childhood cerebellar astrocytoma, childhood brain astrocytoma), basal cell carcinoma, skin cancer (non-melanoma), bile duct cancer, extrahepatic bile duct cancer, intrahepatic bile duct cancer, bladder cancer, urinary bladder cancer, brain cancer (e.g., gliomas and glioblastomas (such as brain stem gliomas, gestational trophoblastic tumor gliomas, cerebellar astrocytomas, brain astrocytomas / malignant gliomas, ependymomas, medulloblastomas, supratentorial primitive neuroectodermal tumors, visual pathway and hypothalamic gliomas), breast cancer, bronchial gland cancer, Tumor / Carcinoid, Carcinoid tumor, Nervous system cancer (e.g., CNS cancer, CNS lymphoma), Cervical cancer, Chronic myeloproliferative disorder, Colorectal cancer (e.g., Colon cancer, Rectal cancer), Lymphoid neoplasms, Mycosis fungoides, Sezary syndrome, Endometrial cancer, Esophageal cancer, Extracranial germ cell tumor, Extragonadal germ cell tumor, Extrahepatic bile duct cancer, Eye cancer, Intraocular melanoma, Retinoblastoma, Gallbladder cancer, Gastrointestinal cancer (e.g., Gastric cancer, Small intestine cancer, Gastrointestinal carcinoid, Gastrointestinal stromal tumor (GIST)), Cholangiocarcinoma, Germ cell tumor, Ovarian germ cell tumor, Head and neck cancer, Neuroendocrine tumors, Hodgkin lymphoma, Ann Arbor Stage III and IV childhood non-Hodgkin lymphoma, ROS1-positive refractory non-Hodgkin lymphoma, leukemia, lymphoma, multiple myeloma, hypopharyngeal cancer, intraocular melanoma, eye cancer, islet cell tumor (endocrine pancreas), kidney cancer (e.g., Wilms tumor, renal cell carcinoma), liver cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), ALK-positive anaplastic large lymphoma, ALK-positive advanced malignant solid tumors, Waldenstrom's macroglobulinoma, melanoma, intraocular (eye) melanoma, Merkel cell carcinoma, mesothelioma, squamous neck cancer with occult primary metastasis, multiple endocrine neoplasia (MEN), myelodysplasia syndrome, myelodysplastic / myeloproliferative disorders, nasopharyngeal carcinoma, neuroblastoma, oral cancer (e.g., oral cavity, lip, oral cavity, tongue, oropharynx, larynx, pharynx), ovarian cancer (e.g., ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor), pancreatic cancer, islet cell pancreatic cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, nasopharyngeal carcinoma, pheochromocytoma, pineoblastoma, metastatic anaplastic thyroid cancer, anaplastic thyroid cancer, papillary thyroid cancer, pituitary tumors, plasmacytoma / multiple myeloma, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, uterine cancer (e.g.,Endometrial cancer, uterine sarcoma, uterine corpus cancer), squamous cell carcinoma, testicular cancer, thymoma, thymic carcinoma, thyroid cancer, juvenile xanthogranuloma, transitional cell carcinoma of the renal pelvis and ureter and other urinary organs, urethral cancer, gestational trophoblastic tumor, vaginal cancer, vulvar cancer, hepatoblastoma, rhabdoid tumor and Wilms tumor.

[0256] Sarcomas treatable with the compounds of the invention include soft tissue cancers and bone cancers, representative examples of which include osteosarcoma or osteogenic sarcoma (bone) (e.g., Ewing's sarcoma), chondrosarcoma (cartilage), leiomyosarcoma (smooth muscle), rhabdomyosarcoma (skeletal muscle), mesothelioma or mesothelioma (membranous lining of body cavities), fibrosarcoma (fibrous tissue), angiosarcoma or hemangioendothelioma (blood vessels), liposarcoma (fatty tissue), glioma or astrocytoma (neurogenic connective tissue found in the brain), myxosarcoma (primitive embryonic connective tissue), mesenchymal or mixed mesodermal tumors (mixed connective tissue type), and histiocytic sarcoma (immune cancer).

[0257] In some embodiments, the methods of the invention are capable of treating subjects suffering from cell proliferative diseases or disorders of the hematologic, liver, brain, lung, colon, pancreas, prostate, ovary, breast, skin, and endometrium.

[0258] As used herein, "hematologic cell proliferative diseases or disorders" (also referred to as "hematologic cancers") include lymphomas, leukemias, myeloid neoplasms, mast cell neoplasms, myelodysplasia, benign monoclonal gammopathy, lymphomatoid papulosis, polycythemia vera, chronic myeloid leukemia, idiopathic myeloid metaplasia, and essential thrombocythemia. Thus, representative examples of hematologic cancers may include multiple myeloma, lymphoma (including T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma (diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), and ALK+ anaplastic large cell lymphoma (e.g., a B-cell non-Hodgkin's lymphoma selected from diffuse large B-cell lymphoma (e.g., germinal center B-cell-like diffuse large B-cell lymphoma or activated B-cell-like diffuse large B-cell lymphoma), Burkitt's lymphoma / leukemia, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, The invention relates to pancreatic cancer, refractory B-cell non-Hodgkin's lymphoma, lymphoplasmacytic lymphoma / Waldenstrom's macroglobulinemia, metastatic pancreatic cancer, refractory B-cell non-Hodgkin's lymphoma, and relapsed B-cell non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas of lymphocytic and cutaneous origin, e.g., small lymphocytic lymphoma, leukemias (including childhood leukemias), hairy cell leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute myeloid leukemia (e.g., acute monocytic leukemia), chronic lymphocytic leukemia, small lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, and mast cell leukemia, myeloid tumors, and mast cell tumors.

[0259] As used herein, "cell proliferative diseases or disorders of the liver" include all forms of cell proliferative disorders that affect the liver. Cell proliferative disorders of the liver can include liver cancer (e.g., hepatocellular carcinoma, intrahepatic cholangiocarcinoma, and hepatoblastoma), precancerous or precancerous conditions of the liver, benign growths or lesions of the liver, malignant growths or lesions of the liver, and metastatic lesions of tissues and organs in the body other than the liver. Cell proliferative disorders of the liver may include hyperplasia, metaplasia, and dysplasia of the liver.

[0260] As used herein, "cell proliferative diseases or disorders of the brain" include all forms of cell proliferative disorders that affect the brain. Cell proliferative disorders of the brain can include brain cancer (e.g., gliomas, glioblastomas, meningiomas, pituitary adenomas, vestibular schwannomas, and primitive neuroectodermal tumors (medulloblastomas)), precancerous or precancerous conditions of the brain, benign growths or lesions of the brain, malignant growths or lesions of the brain, and metastatic lesions of body tissues and organs other than the brain. Cell proliferative disorders of the brain may include hyperplasia, metaplasia, and dysplasia of the brain.

[0261] As used herein, "cell proliferative diseases or disorders of the lung" include all forms of cell proliferative disorders that affect lung cells. Cell proliferative disorders of the lung include lung cancer, precancerous and precancerous conditions of the lung, benign growths or lesions of the lung, hyperplasia, metaplasia and dysplasia of the lung, and metastatic lesions of body tissues and organs other than the lung. Lung cancer includes all forms of lung cancer, for example, malignant lung tumors, carcinoma in situ, typical carcinoid tumors and atypical carcinoid tumors. Lung cancer includes small cell lung cancer ("SLCL"), non-small cell lung cancer ("NSCLC"), adenocarcinoma, small cell carcinoma, large cell carcinoma and mesothelioma. Lung cancer may include "scar cancer", bronchiolar carcinoma, giant cell carcinoma, spindle cell carcinoma and large cell neuroendocrine carcinoma. Lung cancer also includes lung tumors with histological and ultrastructural heterogeneity (e.g., mixed cell types). In some embodiments, the compounds of the present invention can be used to treat non-metastatic or metastatic lung cancer (e.g., NSCLC, ALK-positive NSCLC, NSCLC containing ROS1 rearrangement, lung adenocarcinoma and squamous cell carcinoma).

[0262] As used herein, "cell proliferative diseases or disorders of the colon" include all forms of cell proliferative disorders that affect colon cells, including colon cancer, precancerous or premalignant conditions of the colon, adenomatous polyps of the colon, and metachronous lesions of the colon. Colon cancer includes sporadic and hereditary colon cancer, malignant colon tumors, carcinoma in situ, typical and atypical carcinoid tumors, adenocarcinomas, squamous cell carcinomas, and squamous cell carcinomas. Colon cancer may be associated with hereditary syndromes, such as hereditary nonpolyposis colorectal cancer, common adenomatous polyposis, MYH-associated polyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndrome, and juvenile polyposis. Cell proliferative disorders of the colon may also be characterized by hyperplasia, metaplasia, or dysplasia of the colon.

[0263] As used herein, "cell proliferative diseases or disorders of the pancreas" include all forms of cell proliferative disorders that affect pancreatic cells. Cell proliferative diseases of the pancreas may include pancreatic cancer, precancerous or premalignant conditions of the pancreas, hyperplasia of the pancreas, dysplasia of the pancreas, benign growths or lesions of the pancreas, malignant growths or lesions of the pancreas, and metastatic lesions of body tissues and organs other than the pancreas. Pancreatic cancer includes all forms of pancreatic cancer, including ductal adenocarcinoma, adenosquamous carcinoma, pleomorphic giant cell carcinoma, mucinous adenocarcinoma, osteoclast-like giant cell carcinoma, mucinous cystadenocarcinoma, acinar carcinoma, unclassified large cell carcinoma, small cell carcinoma, pancreatoblastoma, papillary tumors, mucinous cystadenoma, papillary cystic tumors, and serous cystadenoma, as well as pancreatic tumors with histological and ultrastructural heterogeneity (e.g., mixed cells).

[0264] As used herein, "cell proliferative diseases or disorders of the prostate" include all forms of cell proliferative disorders that affect the prostate. Cell proliferative disorders of the prostate can include prostate cancer, precancerous or precancerous conditions of the prostate, benign growths or lesions of the prostate, malignant growths or lesions of the prostate, and metastatic lesions of tissues and organs in the body other than the prostate. Cell proliferative diseases of the prostate may include prostatic hyperplasia, metaplasia, and dysplasia.

[0265] As used herein, "cell proliferative diseases or disorders of the ovary" include all forms of cell proliferative disorders that affect ovarian cells. Cell proliferative disorders of the ovary can include precancerous or precancerous conditions of the ovary, benign growths or lesions of the ovary, ovarian cancer, and metastatic lesions of tissues and organs in the body other than the ovary. Cell proliferative disorders of the ovary may include ovarian hyperplasia, metaplasia, and dysplasia.

[0266] As used herein, "cell proliferative diseases or disorders of the breast" include all forms of cell proliferative disorders that affect breast cells. Cell proliferative disorders of the breast can include breast cancer, precancerous or precancerous conditions of the breast, benign growths or lesions of the breast, and metastatic lesions in body tissues and organs other than the breast. Cell proliferative disorders of the breast may include mammary hyperplasia, metaplasia, and dysplasia.

[0267] As used herein, "cell proliferative diseases or disorders of the skin" include all forms of cell proliferative disorders that affect skin cells. Cell proliferative disorders of the skin can include precancerous or precancerous conditions of the skin, benign growths or lesions of the skin, melanoma, malignant melanoma, or other malignant growths or lesions of the skin, and metastatic lesions in body tissues and organs other than the skin. Cell proliferative disorders of the skin may include skin hyperplasia, metaplasia, and dysplasia.

[0268] As used herein, "cell proliferative diseases or disorders of the endometrium" include all forms of cell proliferative disorders that affect endometrial cells. Cell proliferative disorders of the endometrium can include precancerous or precancerous conditions of the endometrium, benign growths or lesions of the endometrium, endometrial cancer, and metastatic lesions of tissues and organs in the body other than the endometrium. Cell proliferative disorders of the endometrium may include hyperplasia, metaplasia, and dysplasia of the endometrium.

[0269] In some embodiments, the bispecific compound or pharmaceutically acceptable salt or stereoisomer of the invention is a disease or condition of high risk neuroblastoma (NB).

[0270] In some embodiments, the disease or condition that can be treated with the bispecific compounds of the invention is acute myeloid leukemia (AML), multiple myeloma (MM), melanoma, rhabdomyosarcoma, or diffuse large B-cell lymphoma. In other embodiments, the disease or condition is a small solid tumor. In other embodiments, the disease or condition is colon cancer, rectal cancer, gastric cancer, breast cancer, or pancreatic cancer.

[0271] The bispecific compound of formula (I) can be applied to patients, for example, cancer patients, as a monotherapy or by combination therapy. The treatment can be "front / first line", that is, as the initial treatment for patients who have not received a previous anticancer treatment regimen, alone or in combination with other treatments; or "second line", as a treatment for patients who have undergone a previous anticancer treatment regimen, alone or in combination with other treatments; or as a "third line", "fourth line" and other treatments, alone or in combination with other treatments. Patients who have previously received unsuccessful or partially successful treatments but have become unresponsive or intolerant to a particular treatment can also be treated. Treatment can also be given as an adjuvant therapy, that is, to prevent the recurrence of cancer in patients with currently undetectable disease or after surgical removal of the tumor. Therefore, in some embodiments, the compound can be given to patients who have received another treatment (e.g., chemotherapy, radioimmunotherapy, surgical treatment, immunotherapy, radiotherapy, targeted therapy, or any combination thereof).

[0272] The methods of the present invention may require administration of a bispecific compound of Formula (I) or a pharmaceutical composition thereof to a patient in a single dose or multiple doses (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 10, 15, 20 or more doses). For example, the frequency of administration may be from once a day to approximately once every eight weeks. In some embodiments, the frequency of administration ranges from about once a day for 1, 2, 3, 4, 5 or 6 weeks, and in other embodiments requires a 28-day cycle, which includes 3 weeks (21 days) of daily administration. In other embodiments, the bispecific compound may be administered twice a day (BID) (a total of 5 doses) over the course of two and a half days, or once a day (QD) (a total of 2 doses) over the course of two days. In other embodiments, the bispecific compound may be administered once a day (QD) over five days.

[0273] Combination therapy

[0274] The bispecific compound of formula (I) can be combined or used simultaneously with at least one other active agent (e.g., anticancer agent) or therapy for the treatment of diseases and disorders. The terms "combination" and "simultaneous" herein refer to the co-administration of medicaments, which include substantially simultaneous administration, by the same or separate dosage form, and by the same or different modes of administration, or sequential administration, for example, as part of the same treatment regimen, or by a continuous treatment regimen. Therefore, if continuously administered, at the beginning of administration of the second compound, in some cases, the first of the two compounds can still be detected at an effective concentration at the treatment site. The order and time interval can be determined so that they can work together (e.g., synergistically to provide an increased benefit than when administered in other ways). For example, therapeutic agents can be administered simultaneously or sequentially in any order at different time points; however, if not administered simultaneously, they can be administered within a sufficiently close time to provide a desired therapeutic effect, which can be a synergistic approach. Therefore, these terms are not limited to administering active agents at exactly the same time.

[0275] In some embodiments, the treatment regimen may include administering a bispecific compound of Formula (I) in combination with one or more other therapeutic agents known to treat a disease or condition (e.g., cancer). The dose of the additional anticancer therapeutic agent may be the same as or even lower than the known or recommended dose. See, Hardman et al., eds., Goodman & Gilman's The Pharmacological Basis Of Basis Of Therapeutics, 10th ed., McGraw-Hill, New York, 2001; Physician's Desk Reference, 60th ed., 2006. For example, anticancer agents suitable for use in combination with the bispecific compounds of the present invention are known in the art. See, for example, U.S. Pat. No. 9,101,622 (Section 5.2 therein) and U.S. Pat. No. 9,345,705 B2 (Columns 12-18 therein). Representative examples of additional active agents and treatment regimens include radiation therapy, chemotherapy (e.g., mitotic inhibitors, angiogenesis inhibitors, anti-hormones, autophagy inhibitors, alkylating agents, intercalating antibiotics, growth factor inhibitors, anti-androgens, signal transduction pathway inhibitors, anti-microtubule agents, platinum coordination complexes, HDAC inhibitors, proteasome inhibitors, and topoisomerase inhibitors), immunomodulators, therapeutic antibodies (e.g., monospecific and bispecific antibodies), and CAR-T therapy.

[0276] In some embodiments, the bispecific compound of Formula (I) and the additional (e.g., anticancer) therapeutic agent can be administered less than 5 minutes apart, less than 30 minutes apart, less than 1 hour apart, about 1 hour apart, about 1 to about 2 hours apart, about 2 hours to about 3 hours apart, about 3 hours to about 4 hours apart, about 4 hours to about 5 hours apart, about 5 hours to about 6 hours apart, about 6 hours to about 7 hours apart, about 7 hours to about 8 hours apart, about 8 hours to about 9 hours apart, about 9 hours to about 10 hours apart, about 10 hours to about 11 hours apart, about 11 hours to about 12 hours apart, about 12 hours to 18 hours apart, 18 hours to 24 hours apart, 24 hours to 36 hours apart, 36 hours to 48 hours apart, 48 hours to 52 hours apart, 52 hours to 60 hours apart, 60 hours to 72 hours apart, 72 hours to 84 hours apart, 84 hours to 96 hours apart, or 96 hours to 120 hours apart. Two or more (eg, anti-cancer) therapeutic agents may be administered during the same patient visit.

[0277] In some embodiments related to cancer treatment, the bispecific compound of formula (I) and another anticancer agent or therapeutic agent are administered in cycles. Cyclic therapy includes administering an anticancer therapeutic agent for a period of time, followed by administering a second anticancer therapeutic agent for a period of time, and repeating this sequential administration (i.e., the cycle) to reduce the development of resistance to one or both anticancer therapies, avoid or reduce the side effects of one or both anticancer therapies, and / or improve the efficacy of therapy. In one example, cyclic therapy includes administering a first anticancer therapeutic agent for a period of time, followed by administering a second anticancer therapeutic agent for a period of time, optionally, then administering a third anticancer therapeutic agent for a period of time, etc., and repeating this sequential administration (i.e., the cycle) to reduce the development of resistance to one of the anticancer agents, avoid or reduce the side effects of one of the anticancer agents, and / or improve the efficacy of the anticancer agent.

[0278] Medical test kit

[0279] The bispecific compounds of the present invention and / or compositions comprising them can be assembled into a kit or pharmaceutical system. The kit or pharmaceutical system according to this aspect of the present invention includes a carrier or packaging, such as a box, carton, tube, etc., in which one or more containers with strict confinement, such as a vial, tube, ampoule or bottle, contain the bispecific compound of formula (I) of the present invention or its pharmaceutical composition. The kit or pharmaceutical system of the present invention may also include printed instructions for using the compounds and compositions.

[0280] These and other aspects of the invention will be further understood upon consideration of the following examples, which are intended to illustrate certain specific embodiments of the invention, but are not intended to limit the scope of the invention, which is defined by the claims.

[0281] Example

[0282] Example 1: N1-(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4, 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N4-(2-(2-(2-(2-(2,6-dioxapiperidin-3-yl)-1,3- Synthesis of p-phenylenediamine (1).

[0283]

[0284] 5-(tert-Butyl)1-ethyl 3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate

[0285] At 0 ℃, to tert-butyl 3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (4g, 16mmol) and N,N-diisopropylethylamine (DIEA) (5.2mL, 32mmol) in THF (160mL) solution, ethyl chloroformate (1.5mL, 16mmol, dissolved in 40mLTHF) was added dropwise for 30 minutes. Then the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated and then diluted with some water. The mixture was extracted with ethyl acetate (EA). The organic layer was concentrated in vacuo and then purified by silica gel column chromatography (EA / hexane, 40%) to obtain the desired compound (1.6g, 33%) as a white solid.

[0286] LCMS: 325 [M+H] + .

[0287]

[0288] 5-(tert-Butyl)1-ethyl 3-(4-(methoxycarbonyl)benzamido)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate

[0289] To a solution of 5-(tert-butyl)1-ethyl 3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate (972 mg, 3 mmol) and DIEA (990 μL, 6 mmol) in dry DCM (20 mL) at 0° C. was added methyl 4-(chlorocarbonyl)benzoate (713 mg, 3.6 mmol). The mixture was stirred at 40° C. overnight. After the reaction was complete, the mixture was concentrated in vacuo and then purified by silica gel column chromatography (EA / hexane, 40%) to give the desired compound (1.28 g, 87%).

[0290] LCMS: 487 [M+H] + .

[0291]

[0292] 3-(4-(Methoxycarbonyl)benzamido)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester

[0293] At 0 ℃, TFA (2mL) was added to a DCM (8mL) solution of 5-(tert-butyl)1-ethyl 3-(4-(methoxycarbonyl)benzamide)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate (1275mg, 2.6mmol). The mixture was then stirred at room temperature for 2 hours. After the reaction was complete, the mixture was diluted with some water. Before extraction with isopropyl alcohol (IPA) / chloroform (v / v=1 / 3), Et3N (2mL) was added to the mixture. The collected organic layer was concentrated in vacuo and purified by silica gel column chromatography (MeOH / DCM, 16%) to obtain the desired compound (940mg, 72%).

[0294] 1 H NMR (500MHz, DMSO-d6) δ11.03(s,1H),9.78(s,1H),8.19(d,J=8.5Hz,2H),8.06(d,J=8.5Hz ,2H),4.63(s,2H),4.49(q,J=7.1Hz,2H),3.92(s,3H),1.68(s,6H),1.38(t,J=7.1Hz,3H).

[0295] LCMS: 387 [M+H] + .

[0296]

[0297] (S)-5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-3-(4-(methoxycarbonyl)benzamido)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester

[0298] At 0℃, to (S)-N 1 ,N 14-nitrophenyl chloroformate (542 mg, 2.69 mmol) was added to a solution of -dimethyl-2-phenylethane-1,2-diamine (401 mg, 2.45 mmol) and DIEA (808 μL, 4.9 mmol) in dry THF (20 mL). The mixture was stirred at room temperature for 1 hour, and then 3-(4-(methoxycarbonyl)benzamido)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester (850 mg, 2.2 mmol) was added. The mixture was stirred at 60°C overnight. After the reaction was completed, the mixture was concentrated and then purified by silica gel column chromatography (MeOH / DCM, 5%) to obtain the desired compound (498 mg, 39%).

[0299] LCMS: 577 [M+H] + .

[0300]

[0301] (S)-4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzoic acid

[0302] To a solution of (S)-5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-3-(4-(methoxycarbonyl)benzamido)-6,6-dimethyl-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester (498 mg, 0.86 mmol) in MeOH / THF (4 mL, 1:1) was added LiOH (1 M aq., 4.3 mL, 4.3 mmol). The mixture was stirred for 0.5 h. After completion of the reaction, HCl (2 M aq., 2 mL) was added and the mixture was concentrated in vacuo. The crude product was purified by silica gel column chromatography (0.5% TFA in MeOH / DCM, 40%) to give the desired compound (407 mg, 78%) as a TFA salt.

[0303] 1H NMR (500MHz, DMSO-d6) δ12.75(s,1H),11.09(s,1H),10.23(s,1H),8.15-7.93(m,4H),7.51-7.41(m,2H),7.36(t,J=7.7Hz,2H),7.31-7.20(m,1H) ),6.87(s,1H),5.19-5.04(m,1H),4.62(q,J=12.1Hz,2H),3.13(d,J=14. 8Hz, 1H), 2.79 (d, J = 12.0Hz, 1H), 2.49 (s, 6H), 1.65 (s, 3H), 1.58 (s, 3H).

[0304] LCMS: 491[M+H] + .

[0305]

[0306] To a solution of (S)-4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzoic acid (22 mg, 0.036 mmol) and DIEA (60 μL, 0.36 mmol) in dry DCM (2 mL) was added propionic anhydride (50% wt in EA, 115 μL, 0.18 mmol). The reaction was stirred for 10 min before the addition of 4-((2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (16 mg, 0.036 mmol). The mixture was stirred for 30 min and then concentrated in vacuo. The crude product was purified by pre-HPLC to give Compound 1 (2.6 mg, 7%) as a TFA salt.

[0307] 1H NMR (500MHz, DMSO-d6) δ12.47(s,1H),11.10(s,1H),11.02(s,1H),8.66(s,1H),8.09(d,J=8.1Hz,2H),7.95(d,J=8.1Hz,2H),7.58(dd,J=8. 6,7.1Hz,1H),7.38(d,J=7.2Hz,2H),7.30(t,J=7.5Hz,2H),7.20(t,J=7.3Hz,1H),7.14(d,J=8.6Hz,1H),7.04(d,J=7.0Hz,1H),6.60(t,J=5. 8Hz,1H),6.27(s,1H),5.06(dd,J=12.8,5.4Hz,1H),4.90(s,1H),4.57(s,2H),3.61(t,J=5.4Hz,2H),3.54(d,J=3.5Hz,9H),3.45(dt,J=11.2 ,5.6Hz,4H),3.33(s,6H),2.88(ddd,J=16.7,13.7,5.4Hz,1H),2.62-2.53(m,1H),2.22(s,5H),2.10-1.98(m,1H),1.66(s,3H),1.58(s,3H).

[0308] LCMS: 921[M+H] + .

[0309] Example 2: N1-(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4, 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (2).

[0310]

[0311] According to the synthetic route of compound 1 and 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 2 (20.4 mg, 65%) was obtained.

[0312] 1H NMR (500MHz, DMSO-d6) δ11.09(d,J=6.4Hz,2H),9.05(s,1H),8.86(t,J=5.3Hz,1H),8.10(d,J=8.5Hz,2H),7.96(d,J=8.5Hz,2H),7.59(dd,J=8.6,7 .0Hz,1H),7.48-7.43(m,2H),7.40(t,J=7.7Hz,2H),7.34-7.28(m,1H),7. 26(d,J=8.7Hz,1H),7.04(d,J=7.0Hz,1H),6.86(s,1H),6.77(d,J=9.2Hz, 1H),5.36(ddd,J=12.6,9.2,3.9Hz,1H),5.06(dd,J=12.8,5.5Hz,1H),4. 78(d,J=11.8Hz,1H),4.58(d,J=11.9Hz,1H),3.62-3.47(m,5H),3.36(ddd ,1H),2.89(d,J=4.7Hz,3H),2.85(d,J=4.8Hz,3H),2.60(dt,J=17.6,3.5H z, 1H), 2.03 (dtd, J = 11.3, 6.3, 5.6, 3.0Hz, 1H), 1.69 (s, 3H), 1.60 (s, 3H).

[0313] LCMS: 789[M+H] + .

[0314] Example 3: N1-(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4, 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N4-(4-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 4-(4-amino)-4-oxobutyl)terephthalamide (3).

[0315]

[0316] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(4-aminobutyramido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 3 (13.2 mg, 35%) was obtained.

[0317] 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),9.00(s,2H),8.64(t,J=5.5Hz,1H),8.38(d ,J=7.8Hz,1H),8.22-8.05(m,2H),8.04-7.94(m,2H),7.90(d,J=9.3Hz,1H),7.44 (dd,J=8.5,2.0Hz,4H),7.40(td,J=8.3,7.9,2.8Hz,4H),7.36-7.27(m,1H),6.77 (d,J=9.1Hz,1H),5.40-5.31(m,2H),4.93(t,J=7.2Hz,1H),4.79(d,J=11.8Hz,1H) ,4.56(dd,J=17.6,10.6Hz,2H),4.44(t,J=8.0Hz,1H),4.30(s,1H),3.67-3.51(m ,4H),3.39-3.26(m,4H),2.89(d,J=4.8Hz,3H),2.85(d,J=4.8Hz,3H),2.46(s,3H ),2.34(dt,J=14.8,7.8Hz,1H),2.22(dt,J=14.5,7.3Hz,1H),2.01(d,J=14.5Hz, 1H), 1.79 (tdd, 3H), 1.69 (s, 3H), 1.60 (s, 3H), 1.38 (d, J = 6.9Hz, 3H), 0.96 (s, 9H).

[0318] LCMS: 1002 [M+H] + .

[0319] Example 4: N1-(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4, 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (4).

[0320]

[0321] According to the synthetic route of compound 1 and 4-((6-aminohexyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 4 (11 mg, 38%) was obtained.

[0322] 1H NMR (500MHz, DMSO-d6) δ12.55(s,1H),11.07(d,J=21.1Hz,2H),8.64(t,J=5.6Hz,1H),8.13-8.05(m,2H),7.97(d,J=8.0Hz,2H),7.58(dd,J=8.6,7.1 Hz,1H),7.43(d,J=7.2Hz,2H),7.37(t,J=7.6Hz,2H),7.27(t,J=7.3Hz,1H ),7.10(d,J=8.6Hz,1H),7.03(d,J=7.0Hz,1H),6.61(s,1H),6.55(t,J=5. 9Hz,1H),5.21(s,1H),5.06(dd,J=12.7,5.5Hz,1H),4.74(d,J=12.0Hz,1H ),4.57(d,J=11.8Hz,1H),3.34(s,6H),3.29(p,J=6.6Hz,4H),2.89(ddd,J =16.8,13.7,5.4Hz,1H),2.72-2.55(m,6H),2.03(dtd,J=13.1,5.3,2.2Hz ,1H),1.68(s,3H),1.60(s,3H),1.55(m,2H),1.39(dt,J=7.0,3.7Hz,4H).

[0323] LCMS: 845 [M+H] + .

[0324] Example 5: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (5)

[0325]

[0326] According to the synthetic route of compound 1 and 4-((14-amino-3,6,9,12-tetrahydrotetradecyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 5 (5 mg, 17%) was obtained.

[0327] 1H NMR (500MHz, DMSO-d6) δ12.49(s,1H),11.10(s,1H),11.03(s,1H),8.67(s,1H),8.09(d,J=8.1Hz,2H),7.96(d,J=8.1Hz,2H),7.58(dd,J=8.6 ,7.0Hz,1H),7.39(d,J=7.2Hz,2H),7.32(t,J=7.5Hz,2H),7.22(t,J=7.3Hz,1H),7.14(d,J=8.6Hz,1H),7.04(d,J=7.0Hz,1H),6.60(t,J=5.8 Hz,1H),6.35(s,1H),5.06(dd,J=12.7,5.4Hz,1H),4.98(s,1H),4.58(s,2H),3.62(t,J=5.4Hz,2H),3.57-3.49(m,12H),3.46(dt,J=12.0,5. 8Hz,5H),2.89(ddd,J=16.9,13.7,5.4Hz,1H),2.63-2.54(m,2H),2.31(s,1H),2.03(ddd,J=11.7,6.4,3.9Hz,1H),1.66(s,3H),1.59(s,3H).

[0328] LCMS: 965 [M+H] + .

[0329] Example 6: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)- 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane- Synthesis of terephthalamide (6):

[0330]

[0331] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(3-(2-(2-aminoethoxy)ethoxy)propionamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 6 (6.5 mg, 20%) was obtained.

[0332] 1H NMR(500MHz,DMSO-d6)δ11.08(s,1H),9.05(s,1H),8.99(s,1H),8.69(t,J=5.6Hz,1H),8 .38(d,J=7.8Hz,1H),8.10(d,J=8.5Hz,2H),7.98(d,J=8.5Hz,2H),7.87(d,J=9.3Hz,1H), 7.46-7.42(m,4H),7.42-7.35(m,4H),7.33-7.29(m,1H),6.77(d,J=9.1Hz,1H),5.36(dd d,J=12.4,9.1,4.0Hz,2H),4.91(p,J=7.3Hz,2H),4.79(d,J=11.9Hz,1H),4.55(dd,J=19. 3,10.6Hz,2H),4.43(t,J=8.1Hz,1H),4.29(dd,J=4.7,2.4Hz,1H),3.67-3.47(m,11H),3 .45(q,J=6.8,6.3Hz,2H),3.36(ddt,J=12.6,8.9,4.4Hz,1H),2.89(d,J=4.8Hz,3H),2.84 (d,J=4.8Hz,3H),2.46(s,3H),2.36(dt,J=14.6,6.1Hz,1H),2.07-1.99(m,1H),1.80(dd d,J=12.9,8.5,4.6Hz,1H),1.69(s,3H),1.60(s,3H),1.37(d,J=7.0Hz,3H),0.94(s,9H).

[0333] LCMS: 1076 [M+H] + .

[0334] Example 7: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (7).

[0335]

[0336] According to the synthetic route of compound 1 and 4-(6-aminohex-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 7 (3.3 mg, 12%) was obtained.

[0337] 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),11.13(s,1H),11.04(s,1H),9.71(s,1H),8.73(s,1H),8.11(d,J=8.1Hz,2H),7.98(d, J=8.1Hz,2H),7.90-7.85(m,1H),7.84-7.81(m,2H),7.45(d,J=7.7Hz,2H),7.38(t,J=7.6Hz,2H),7.29(t,J=7.3Hz,1H),6.7 3(s,1H),5.32(s,1H),5.14(dd,J=12.8,5.5Hz,1H),4.81(d,J=12.1Hz,1H),4.57(d,J=11.8Hz,1H),2.93-2.70(m,4H),2.60 (t,J=7.0Hz,4H),2.09-2.00(m,1H),1.76(q,J=7.4,6.7Hz,2H),1.68(d,J=6.3Hz,4H),1.60(s,3H),1.21(d,J=29.7Hz,2H).

[0338] LCMS: 826 [M+H] + .

[0339] Example 8: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (8).

[0340]

[0341] According to the synthetic route of compound 1 and 4-((5-aminopentyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 8 (7.2 mg, 29%) was obtained.

[0342] 1H NMR (500MHz, DMSO-d6) δ11.04(d,J=18.1Hz,2H),8.59(t,J=5.7Hz,1H),8.04(d,J=8.3Hz,2H),7.91(d,J=8.1Hz,2H),7.76(dd,J=8.5,7.2Hz,1H),7. 48(d,J=8.6Hz,1H),7.38(dd,J=14.5,7.4Hz,3H),7.30(t,J=7.6Hz,2H),7 .21(t,J=7.2Hz,1H),6.51(s,1H),5.03(dd,J=12.8,5.5Hz,1H),4.64(s,1H ),4.52(d,J=11.9Hz,1H),4.18(t,J=6.4Hz,1H),3.55(s,1H),3.31-3.26( m,6H),3.07(s,1H),3.01(q,J=7.2Hz,3H),2.83(ddd,J=17.0,13.8,5.5Hz ,2H),2.59-2.49(m,3H),2.01-1.95(m,1H),1.77(p,J=6.6Hz,2H),1.62(s ,3H),1.58(t,J=7.4Hz,2H),1.54(s,3H),1.48(qd,J=9.9,9.0,6.0Hz,2H).

[0343] LCMS: 832 [M+H] + .

[0344] Example 9: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (9).

[0345]

[0346] According to the synthetic route of compound 1 and 4-((8-aminooctyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 9 (15 mg, 52%) was obtained.

[0347] 1H NMR (500MHz, DMSO-d6) δ12.49(s,1H),11.11(s,1H),11.02(s,1H),8.60(t,J=5.7Hz,1H),8.09(d,J=8.1Hz,2H),7.95(d,J=8.1Hz,2H),7.81( dd,J=8.5,7.3Hz,1H),7.52(d,J=8.5Hz,1H),7.44(d,J=7.2Hz,1H),7.32(t,J=7.6Hz,2H),7.22(t,J=7.3Hz,1H),6.33(s,1H),5.09(dd,J=12. 8,5.5Hz,1H),4.95(s,1H),4.59(s,2H),4.21(t,J=6.4Hz,2H),3.33(s,6H),2.89(ddd,J=16.8,13.8,5.4Hz,1H),2.64-2.54(m,1H),2.33-2.1 8(m,5H),2.03(dtd,J=13.0,5.3,2.2Hz,1H),1.83-1.71(m,2H),1.66( s, 3H), 1.63-1.53 ​​(m, 5H), 1.48 (t, J = 7.8Hz, 2H), 1.35 (d, J = 4.1Hz, 6H).

[0348] LCMS: 874 [M+H] + .

[0349] Example 10: N-((S)-2-(dimethylamino)-1-phenylethyl)-3-(4-(11-((2-(2,6-dioxopiperidine-3- 1,3-dihydroisoindolin-4-yl)oxy)undecanoyl)benzamido)-6,6-dimethyl-4,6-dihydropyrrolidone Synthesis of pyrrol[3,4-c]pyrazole-5(1H)-carboxamide (10).

[0350]

[0351] 5-(tert-Butyl)1-ethyl 6,6-dimethyl-3-(4-nitrobenzamide)-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate

[0352] To a solution of 5-(tert-butyl)1-ethyl 3-amino-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate (875 mg, 2.7 mmol) and DIEA (1.34 mL, 8.1 mmol) in DCM (40 mL) was added 4-nitrobenzoyl chloride (600 mg, 3.24 mmol) at 0°C. The reaction was stirred at room temperature overnight. The mixture was concentrated in vacuo and then purified by silica gel column chromatography (EA / hexane, 30%) to give the desired compound (822 mg, 64%).

[0353] LCMS: 474 [M+H] + .

[0354]

[0355] 6,6-Dimethyl-3-(4-nitrobenzamido)-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester

[0356] To a solution of 5-(tert-butyl)-1-ethyl-6,6-dimethyl-3-(4-nitrobenzamide)-4,6-dihydropyrrolo[3,4-c]pyrazole-1,5-dicarboxylate (822 mg, 1.74 mmol) in DCM (8 mL) was added TFA (2 mL) at 0°C. The mixture was stirred at room temperature for 2 hours and then concentrated in vacuo. The crude product was used in the next step without further purification.

[0357] LCMS: 374 [M+H] + .

[0358]

[0359] (S)-5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-3-(4-nitrobenzamide)-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester

[0360] To a solution of ethyl 6,6-dimethyl-3-(4-nitrobenzamido)-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylate (633 mg, 1.3 mmol) and DIEA (130 μL, 0.78 mmol) in DCM (10 mL) was added triphosgene (194 mg, 0.65 mmol, dissolved in 2 mL DCM) dropwise at 0°C. The mixture was stirred at 0°C for 30 minutes. Then (S)-N 1 ,N 1 -dimethyl-2-phenylethane-1,2-diamine (256 mg, 1.56 mmol) and DIEA (320 μL, 1.95 mmol). The mixture was stirred at 40°C overnight. After the reaction was completed, the mixture was concentrated in vacuo and then purified by silica gel column chromatography (MeOH / DCM, 6%) to give the desired compound (232 mg, 32%).

[0361] LCMS: 564 [M+H] + .

[0362]

[0363] (S)-3-(4-aminobenzamido)-N-(2-(dimethylamino)-1-phenylethyl)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide

[0364] To a solution of (S)-5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-3-(4-nitrobenzamido)-5,6-dihydropyrrolo[3,4-c]pyrazole-1(4H)-carboxylic acid ethyl ester (232 mg, 0.41 mmol) in isopropanol (2 mL) was added LiOH (1N aq., 2 mL). The reaction was stirred at room temperature for 10 minutes and then extracted with IPA / chloroform (v / v=1 / 3). The combined organic layers were concentrated in vacuo. The resulting product was then redissolved in MeOH (20 mL). Palladium (10% on activated carbon, 30 mg) was added and the mixture was stirred under H2 atmosphere for 3 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated in vacuo. The crude product was purified by silica gel column chromatography (1.75N NH3 in MeOH / DCM, 30%) to give the desired compound (153 mg, 81%, 2 steps).

[0365] 1 H NMR (500MHz, DMSO-d6) δ12.33(s,0H),10.33(s,0H),9.19(s,0H),7.77(d,J=8.3Hz,1H),7.41(d,J=7.5Hz,1H),6.57(d,J=8.3Hz,1H), 5.88(s,0H),5.75(s,1H),4.63(s,1H),4.53(d,J=11.9Hz,2H),3.17(d,J=5.2Hz,1H),2.64(p,J=1.8Hz,1H),1.66(s,3H),1.58(s,3H).

[0366] LCMS: 462[M+H] + .

[0367]

[0368] To a solution of 11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)undecanoic acid (14 mg, 0.03 mmol) and DIEA (50 μL, 0.3 mmol) in dry DCM (2 mL) was added T3P (50% wt in EA, 90 μL, 0.15 mmol). The reaction was stirred for 10 minutes before the addition of (S)-3-(4-aminobenzamido)-N-(2-(dimethylamino)-1-phenylethyl)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide (14 mg, 0.03 mmol). The mixture was stirred for 30 minutes and then concentrated in vacuo. The crude product was purified by preparative HPLC to give compound 10 (7.6 mg, 25%) as a TFA salt.

[0369] 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),10.79(s,1H),10.16(s,1H),9.00(s,1H),7.98(d, J=8.7Hz,2H),7.81(dd,J=8.5,7.2Hz,1H),7.72(d,J=8.6Hz,2H),7.51(d,J=8.5Hz,1H),7 .44(d,J=7.4Hz,3H),7.42-7.38(m,2H),7.34-7.28(m,1H),6.76(d,J=9.1Hz,1H),5.35(d dd,J=12.4,9.0,3.9Hz,1H),5.08(dd,J=12.8,5.5Hz,1H),4.76(d,J=11.9Hz,1H),4.55(d ,J=11.8Hz,1H),4.20(t,J=6.4Hz,2H),3.59-3.51(m,1H),3.35(ddt,J=17.1,12.3,6.2H z,1H),3.18-3.13(m,1H),2.89(d,J=4.8Hz,3H),2.84(d,J=4.9Hz,3H),2.59(dt,J=17.4, 3.4Hz,1H),2.34(t,J=7.4Hz,2H),2.03(dtd,J=13.0,5.2,2.2Hz,1H),1.75(q,J=6.9,6.4 Hz, 2H), 1.68 (s, 3H), 1.59 (s, 3H), 1.46 (dq, J = 15.1, 7.3, 6.7Hz, 2H), 1.38-1.22 (m, 12H).

[0370] LCMS: 902 [M+H] + .

[0371] Example 11N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4- 5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (11).

[0372]

[0373] According to the synthetic route of compound 1 and 4-((4-aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 9 (15 mg, 52%) was obtained.

[0374] LCMS: 817 [M+H] + .

[0375] Example 12: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(6-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 6-amino)-6-oxyhexyl)terephthalamide (12).

[0376]

[0377] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(6-aminohexanamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 12 (8.6 mg, 28%) was obtained.

[0378] 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),9.00(s,1H),8.60(t,J=5.6Hz,1H),8.38(d,J=7.8Hz,1H ),8.09(d,J=8.5Hz,2H),7.96(d,J=8.5Hz,2H),7.80(d,J=9.3Hz,1H),7.65(s,1H),7.48-7.42 (m,4H),7.42-7.38(m,4H),7.34-7.28(m,1H),6.77(d,J=9.2Hz,1H),5.36(ddd,J=12.5,9.1,3 .9Hz,2H),4.92(t,J=7.2Hz,1H),4.78(d,J=11.8Hz,1H),4.58(d,J=11.9Hz,2H),4.53(d,J=9.3 Hz,1H),4.43(t,J=8.0Hz,1H),4.29(t,J=3.6Hz,1H),3.66-3.50(m,5H),3.35(td,J=8.6,4.2H z,1H),3.27(q,J=6.5Hz,2H),2.89(d,J=4.8Hz,3H),2.85(d,J=4.8Hz,3H),2.46(s,3H),2.27(d q,J=13.3,6.9,6.1Hz,1H),2.15(tdd,J=14.3,8.2,5.5Hz,2H),2.02(ddd,J=11.1,7.8,2.7Hz, 1H), 1.80 (td, J=8.4, 4.2Hz, 1H), 1.69 (s, 3H), 1.60 (s, 3H), 1.38 (d, J=7.0Hz, 3H), 0.94 (s, 9H).

[0379] LCMS: 1030 [M+H] + .

[0380] Example 13: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole Synthesis of terephthalamide (13).

[0381]

[0382] According to the synthetic route of compound 1 and 4-(6-aminohexyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 13 (12 mg, 48%) was obtained.

[0383] 1H NMR(500MHz,DMSO-d6)δ11.12(s,1H),11.08(s,1H),9.05(s,1H),8.60(t,J=5.6 Hz,1H),8.09(d,J=8.5Hz,2H),7.96(d,J=8.5Hz,2H),7.80-7.74(m,2H),7.71(d d,J=6.9,1.9Hz,1H),7.45(d,J=7.3Hz,2H),7.40(t,J=7.7Hz,2H),7.35-7.29(m ,1H),6.77(d,J=9.1Hz,1H),5.36(ddd,J=12.6,9.1,3.9Hz,1H),5.13(dd,J=12.7 ,5.5Hz,1H),4.78(d,J=11.9Hz,1H),4.58(d,J=11.9Hz,1H),3.56(td,J=12.9,2 .7Hz,1H),3.36(ddd,J=12.8,8.8,4.0Hz,1H),3.27(q,J=6.6Hz,2H),3.09-2.99 (m,2H),2.89(d,J=4.7Hz,3H),2.86(s,3H),2.65-2.52(m,2H),2.06(dp,J=10.5 ,3.3Hz,1H),1.69(s,3H),1.60(s,5H),1.55(t,J=7.0Hz,2H),1.42-1.33(m,4H).

[0384] LCMS: 830 [M+H] + .

[0385] Example 14: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(2-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3- 2,3-dihydro-1H-benzo[de]isoquinolin-4-yl)oxy)ethoxy)ethoxy)ethoxy)ethyl)terephthalate Synthesis of amide (14).

[0386]

[0387] According to the synthetic route of compound 1 and 4-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione in Example 1, compound 14 (5 mg, 16%) was obtained.

[0388] 1H NMR(500MHz,DMSO-d6)δ11.07(s,1H),11.02(s,1H),9.01(s,1H),8.67(t,J=5.6Hz,1H),8.46-8.26(m,2H),8.14-8.06(m,2H),8.03-7.93(m,4H), 7.84(dt,J=11.0,7.8Hz,1H),7.48-7.37(m,4H),7.33-7.26(m,1H),6.76 (d,J=9.2Hz,1H),5.83(dt,J=11.9,6.0Hz,1H),5.36(td,J=10.6,9.1,3.9 Hz,1H),4.78(d,J=11.8Hz,1H),4.57(d,J=11.8Hz,1H),4.40-4.21(m,2H ),3.85(d,J=4.2Hz,2H),3.63(dt,J=5.7,2.5Hz,2H),3.56(dt,J=8.2,5.1 Hz,10H),3.44(t,J=5.8Hz,2H),2.89(d,J=4.7Hz,3H),2.84(d,J=4.7Hz, 3H), 2.65-2.54 (m, 2H), 2.06 (d, J = 19.3Hz, 1H), 1.69 (s, 3H), 1.60 (s, 3H).

[0389] LCMS: 972 [M+H] + .

[0390] Example 15: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1- (4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutane-2-yl Synthesis of terephthalamide (15).

[0391]

[0392] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(3-(2-aminoethoxy)propionamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 15 (11.7 mg, 33%) was obtained.

[0393] 1H NMR(500MHz,DMSO-d6)δ11.09(s,1H),8.99(s,2H),8.66(t,J=5.7Hz,1H),8.39(d, J=7.8Hz,1H),8.10(d,J=8.5Hz,2H),7.99(d,J=8.5Hz,2H),7.90(d,J=9.3Hz,1H),7 .47-7.42(m,4H),7.42-7.35(m,4H),7.32-7.27(m,1H),6.77(d,J=9.1Hz,1H),5.3 6(ddd,J=12.6,9.1,3.9Hz,1H),4.91(p,J=7.0Hz,1H),4.78(d,J=11.9Hz,1H),4.56 (dd,J=19.3,10.6Hz,2H),4.44(t,J=8.1Hz,1H),4.29(dd,J=4.7,2.4Hz,1H),3.72 -3.50(m,8H),3.45(q,J=5.5Hz,2H),2.89(d,J=4.8Hz,3H),2.84(d,J=4.8Hz,3H),2 .61-2.53(m,1H),2.46(s,3H),2.42-2.35(m,1H),2.11-1.99(m,1H),1.79(ddd,J=1 3.0, 8.6, 4.5Hz, 1H), 1.69 (s, 3H), 1.60 (s, 3H), 1.36 (d, J = 7.0Hz, 3H), 0.93 (s, 9H).

[0394] LCMS: 1032 [M+H] + .

[0395] Example 16: N-((S)-2-(dimethylamino)-1-phenylethyl)-3-(4-(4-((2-(2,6-dioxopiperidine-3- 1,3-dihydroisoindolin-4-yl)amino)butyrylamide)benzamide)-6,6-dimethyl-4,6-dihydropyrrolo[2-(2-(4-(2-nitro-1-yl)amino)butyrylamide)benzamide)-6,6-dimethyl-4,6-dihydropyrrolo[2-(2-nitro-1-yl)amino)butyrylamide)benzamide)-6,6-dimethyl-4 Synthesis of [3,4-c]pyrazole-5(1H)-carboxamide (16).

[0396]

[0397] According to the synthetic route of compound 10 and 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyric acid in Example 10, compound 16 (9.5 mg, 35%) was obtained.

[0398] LCMS: 803 [M+H] + .

[0399] Example 17: Synthesis of N-((S)-2-(dimethylamino)-1-phenylethyl)-3-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)hex-5-ynylamide)benzamide)-6,6-dimethyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide (17).

[0400]

[0401] According to the synthetic route of compound 10 and 6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)hex-5-ynoic acid in Example 10, compound 17 (5.2 mg, 19%) was obtained.

[0402] 1 H NMR (500MHz, DMSO-d6) δ12.42(s,1H),11.00(s,1H),10.76(s,1H),10.26(s,1H),7.98(d,J=8.4Hz,2H),7.72(dd,J=7.9,3. 5Hz,3H),7.65(t,J=9.1Hz,2H),7.53(t,J=7.6Hz,1H),7.40(d,J=7.6Hz,2H),7.33(t,J=7.5Hz,2H),7.24(d,J=8.3Hz,1H), 5.14(dd,J=13.3,5.1Hz,1H),5.01(s,1H),4.63-4.53(m,1H),4.49(d,J=17.9Hz,1H),4.35(d,J=7.7Hz,1H),2.92(ddd,J=1 7.3, 13.6, 5.4Hz, 1H), 2.58 (q, J = 7.3Hz, 7H), 2.04-1.98 (m, 1H), 1.93 (td, J = 8.4, 7.2, 4.9Hz, 3H), 1.66 (s, 3H), 1.58 (s, 3H).

[0403] LCMS: 798 [M+H] + .

[0404] Example 18: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-3,3-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(6-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 6-amino)-6-oxypropyl)terephthalamide (18).

[0405]

[0406] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(3-aminopropionamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 18 (3.6 mg) was obtained.

[0407] 1 H NMR (500MHz, DMSO-d6) δ12.61(s,1H),11.05(s,1H),8.99(s,1H),8.68(s,1H),8.42(d,J=7.8Hz,1H),8.11(d,J=7.9Hz,2H),7.98(d,J=8. 7Hz,2H),7.50-7.20(m,9H),6.75(s,1H),5.33(s,1H),5.17(s,1H),4.91(q,J=7.4Hz,1H),4.82(d,J=11.8Hz,1H),4.55(dd,J=14.4,10.4H z,2H),4.45(t,J=8.0Hz,1H),4.29(s,1H),3.63(d,J=4.4Hz,2H),3.50(ddd,J=29.3,13.5,6.6Hz,2H),2.79(s,6H),2.59(dt,J=14.7,7.4 Hz,1H),2.46(s,3H),2.09-1.98(m,1H),1.80(ddd,J=12.9,8.5,4.7Hz,1H),1.69(s,3H),1.61(s,3H),1.38(d,J=7.0Hz,3H),0.94(s,9H).

[0408] LCMS: 988 [M+H] + .

[0409] Example 19: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,2-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxopiperidin-3-yl)- Synthesis of terephthalamide (19): (4-(2-isoindolin-4-yl)amino)ethoxy)ethyl)terephthalamide (19).

[0410]

[0411] According to the synthetic route of compound 1 and 4-((2-(2-aminoethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 19 (5.9 mg, 21%) was obtained.

[0412] 1H NMR(500MHz,DMSO-d6)δ12.52(s,1H),11.09(d,J=20.2Hz,2H),9.12(s,1H) ,8.69(t,J=5.6Hz,1H),8.47(s,1H),8.08(d,J=8.3Hz,2H),7.96(d,J=8.0H z,2H),7.57(dd,J=8.6,7.1Hz,1H),7.43(d,J=7.5Hz,2H),7.37(t,J=7.6Hz ,2H),7.29(d,J=7.4Hz,1H),7.17(d,J=8.6Hz,1H),7.03(d,J=7.0Hz,1H),6. 64(t,J=5.8Hz,1H),5.24(s,1H),5.05(dd,J=12.8,5.5Hz,1H),4.72(s,1H) ,4.58(d,J=11.8Hz,1H),3.64(dt,J=26.2,5.6Hz,4H),3.49(dq,J=12.0,5. 7Hz, 4H), 3.16 (d, J=16.7Hz, 2H), 2.89 (ddd, J=17.0, 13.9, 5.4Hz, 2H), 2.65 -2.53(m,1H),2.02(dtd,J=12.6,5.2,2.2Hz,1H),1.68(s,3H),1.60(s,3H).

[0413] LCMS: 833 [M+H] + .

[0414] Example 20: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-3,5-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(6-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 6-amino)-6-oxopentyl)terephthalamide (20).

[0415]

[0416] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(5-aminopentanamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 20 (9.1 mg, 28%) was obtained.

[0417] 1H NMR(500MHz,DMSO-d6)δ12.49(s,1H),11.03(s,1H),8.99(s,1H),8.62(s, 0H),8.38(d,J=7.8Hz,1H),8.10(d,J=8.1Hz,2H),7.95(d,J=8.0Hz,2H),7. 82(d,J=9.3Hz,1H),7.467.42(m,2H),7.41-7.36(m,4H),7.32(t,J=7.7Hz ,2H),7.22(t,J=7.3Hz,1H),6.34(s,1H),5.11(d,J=3.6Hz,1H),4.93(q,J= 7.2Hz,1H),4.59(s,2H),4.53(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.2 9(s,1H),3.70-3.54(m,2H),2.46(s,3H),2.30(q,J=14.3,7.8Hz,4H),2.17 (dd,J=14.0,7.5Hz,2H),2.02(td,J=8.9,4.4Hz,1H),1.81(td,J=8.4,4.2H z, 1H), 1.66 (s, 3H), 1.60-1.48 (m, 7H), 1.38 (d, J = 7.0Hz, 3H), 0.95 (s, 9H).

[0418] LCMS: 1016 [M+H] + .

[0419] Example 21: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxopiperidin-3-yl) Synthesis of isoindolyl-5-yl)amino)pentyl)terephthalamide (21).

[0420]

[0421] According to the synthetic route of compound 1 and 5-((5-aminopentyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 21 (8.5 mg, 30%) was obtained.

[0422] 1H NMR (500MHz, DMSO-d6) δ11.07(d,J=3.8Hz,2H),9.01(s,1H),8.63(t,J=5.7Hz,1H),8.09(d,J=8.2Hz,2H),7.96(d,J=8.5Hz,2H),7.57(d,J=8.4Hz,1 H),7.45(d,J=7.4Hz,2H),7.40(t,J=7.7Hz,2H),7.34-7.27(m,1H),6.96( d,J=2.1Hz,1H),6.86(dd,J=8.4,2.2Hz,1H),6.77(d,J=9.2Hz,1H),5.36( ddd,J=12.5,9.1,3.9Hz,1H),5.03(dd,J=12.7,5.5Hz,1H),4.78(d,J=11. 9Hz,1H),4.58(d,J=11.9Hz,1H),3.64-3.42(m,10H),3.21-3.15(m,2H),2 .89(d,J=4.9Hz,3H),2.85(d,J=4.8Hz,3H),2.58(dt,J=20.4,4.2Hz,1H), 2.06-1.95(m,1H),1.69(s,3H),1.60(s,3H),1.45(td,J=8.4,4.2Hz,2H).

[0423] LCMS: 831[M+H] + .

[0424] Example 22: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(7-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 7-oxoheptyl)terephthalamide (22).

[0425]

[0426] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(7-aminoheptanoylamino)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 22 (6.9 mg, 20%) was obtained.

[0427] 1H NMR(500MHz,DMSO-d6)δ12.47(s,1H),11.02(s,1H),8.99(s,1H),8.59(s,1H),8.3 7(d,J=7.8Hz,1H),8.09(d,J=8.0Hz,2H),7.95(d,J=8.0Hz,2H),7.80(d,J=9.3Hz, 1H),7.46-7.42(m,2H),7.38(d,J=8.1Hz,4H),7.30(t,J=7.6Hz,2H),7.20(t,J=7. 3Hz,1H),6.27(s,1H),5.10(d,J=3.5Hz,1H),4.92(p,J=8.9,8.1Hz,2H),4.57(s,1 H),4.53(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.29(s,1H),3.62(d,J=4.1Hz,2H ),3.27(d,J=6.9Hz,2H),2.46(s,3H),2.27(dd,J=14.3,7.5Hz,1H),2.16-2.11(m, 1H),2.05-1.96(m,1H),1.80(ddd,J=12.8,8.5,4.6Hz,1H),1.66(s,3H),1.58(s,3 H), 1.51 (tt, J = 14.5, 7.2 Hz, 4H), 1.37 (d, J = 7.0 Hz, 3H), 1.31 (s, 4H), 0.94 (s, 9H).

[0428] LCMS: 1044 [M+H] + .

[0429] Example 23: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(8-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of terephthalamide (23).

[0430]

[0431] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(8-aminooctanoylamino)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 23 (12.3 mg, 36%) was obtained.

[0432] 1H NMR(500MHz,DMSO-d6)δ12.53(s,1H),11.09(s,1H),9.20(s,1H),8.99(s,1H),8.62(t,J=5.7Hz,1H), 8.38(d,J=7.8Hz,1H),8.10(d,J=8.4Hz,2H),7.96(d,J=8.1Hz,2H),7.79(d,J=9.3Hz,1H),7.44(dd,J =7.8,5.7Hz,4H),7.42-7.36(m,4H),7.33-7.27(m,1H),6.77(d,J=9.1Hz,1H),5.36(ddd,J=12.5,9.1 ,3.9Hz,1H),5.11(d,J=3.6Hz,1H),4.98-4.88(m,1H),4.80(d,J=11.8Hz,1H),4.57(d,J=11.9Hz,1H), 4.52(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.31-4.24(m,1H),3.61(t,J=3.9Hz,2H),3.55(t,J=12. 5Hz,1H),3.27(q,J=6.7Hz,2H),2.86(d,J=19.3Hz,6H),2.46(s,3H),2.27(dd,J=14.3,7.4Hz,1H),2.1 2(ddd,J=14.0,7.9,6.1Hz,1H),2.02(ddd,J=10.8,7.4,2.7Hz,1H),1.80(ddd,J=12.9,8.4,4.6Hz,1H ),1.69(s,3H),1.60(s,3H),1.57-1.45(m,4H),1.38(d,J=7.0Hz,3H),1.34-1.23(m,4H),0.94(s,9H).

[0433] LCMS: 1058 [M+H] + .

[0434] Example 24: N-((S)-2-(dimethylamino)-1-phenylethyl)-3-(4-((2S,4R)-4-hydroxy-2-(((S)- 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)benzamido)-6,6-dimethyl Synthesis of 4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide (24).

[0435]

[0436] According to the synthetic route of compound 1 and (2S,4R)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide in Example 1, compound 24 (3.6 mg) was obtained.

[0437] 1H NMR (500MHz, DMSO-d6) δ12.49(s,1H),11.05(s,1H),8.99(s,1H),8.51(d,J=7.8Hz,1H),8.10(d,J=8.0Hz,2H),7 .68(d,J=7.7Hz,2H),7.45(d,J=8.3Hz,2H),7.43-7.37(m,4H),7.33(t,J=7.2Hz,2H),7.29-7.20(m,1H),5.05(s, 1H),4.99(q,J=7.2Hz,1H),4.68-4.48(m,2H),4.26(s,1H),3.73(dd,J=10.9,3.7Hz,1H),3.27(d,J=11.0Hz,1H), 2.46(s,3H),2.44(s,1H),2.22-2.15(m,1H),1.93-1.81(m,1H),1.67(s,3H),1.59(s,3H),1.43(d,J=7.0Hz,3H).

[0438] LCMS: 804 [M+H] + .

[0439] Example 25: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -((S)-14-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4- methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-15,15-dimethyl-12-oxo-3,6,9- Synthesis of trioxa-13-azahexadecyl)terephthalamide (25).

[0440]

[0441] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-1-amino-14-(tert-butyl)-12-oxo-3,6,9-trioxa-13-azapentadecan-15-yl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 25 (7.2 mg, 19%) was obtained.

[0442] 1H NMR(500MHz,DMSO-d6)δ11.08(s,1H),9.13-8.92(m,2H),8.69(t,J=5.6Hz, 1H),8.38(d,J=7.8Hz,1H),8.21-8.06(m,3H),8.00-7.96(m,2H),7.86(d,J= 9.3Hz,1H),7.49-7.36(m,9H),7.30(td,J=6.8,6.4,1.5Hz,1H),6.77(d,J= 9.2Hz,1H),5.41-5.29(m,2H),4.92(t,J=7.3Hz,1H),4.78(d,J=11.9Hz,1H) ,4.63-4.48(m,2H),4.43(t,J=8.0Hz,1H),4.28(s,1H),3.69-3.41(m,16H) ,3.38-3.32(m,1H),2.89(d,J=4.8Hz,3H),2.85(d,J=4.8Hz,3H),2.46(s,3H ), 2.35 (dt, J=14.5, 6.1Hz, 1H), 2.02 (dd, J=12.1, 8.7Hz, 1H), 1.80 (td, J=8. 4, 4.3Hz, 1H), 1.69 (s, 3H), 1.60 (s, 3H), 1.38 (d, J = 7.0Hz, 3H), 0.94 (s, 9H).

[0443] LCMS: 1120 [M+H] + .

[0444] Example 26: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,2-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3- Synthesis of (4-(dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)terephthalamide (26).

[0445]

[0446] According to the synthetic route of compound 1 and 4-((2-(2-(2-aminoethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione in Example 1, compound 26 (14.7 mg, 49%) was obtained.

[0447] 1H NMR(500MHz,DMSO-d6)δ11.09(d,J=9.6Hz,2H),9.04(s,1H),8.66(t,J=5.6Hz, 1H),8.09(d,J=8.5Hz,2H),7.97(d,J=8.5Hz,2H),7.58(dd,J=8.6,7.0Hz,1H), 7.47-7.43(m,2H),7.40(dd,J=8.5,6.8Hz,2H),7.34-7.29(m,1H),7.13(d,J=8 .6Hz,1H),7.04(d,J=7.0Hz,1H),6.77(d,J=9.2Hz,1H),6.61(t,J=5.9Hz,1H), 5.36(ddd,J=12.5,9.2,3.8Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),4.78(d,J=11 .9Hz,1H),4.57(d,J=11.9Hz,1H),3.66-3.51(m,10H),3.48-3.37(m,4H),3.36 (ddd,J=12.6,8.6,4.0Hz,1H),2.89(d,J=4.6Hz,3H),2.85(d,J=4.7Hz,3H),2. 63-2.53(m,2H),2.03(ddd,J=10.6,5.5,3.0Hz,1H),1.69(s,3H),1.60(s,3H).

[0448] LCMS: 877[M+H] + .

[0449] Example 27: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo- Synthesis of 2,3-dihydro-1H-benzo[de]isoquinolin-5-yl)oxy)propyl)terephthalamide (27).

[0450]

[0451] According to the synthetic route of compound 1 and 5-(3-aminopropoxy)-2-(2,6-dioxopiperidin-3-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione in Example 1, compound 27 (7.5 mg) was obtained.

[0452] 1H NMR (500MHz, DMSO-d6) δ11.00(s,1H),10.95(s,1H),8.96(s,1H),8.70(q,J=5.5Hz,1H),8.36-8.20(m,2H),8.07-7.99(m,3H),7.97-7.88 (m,4H),7.78(dt,J=10.3,7.8Hz,1H),7.39-7.29(m,4H),7.27-7.20(m,1H),6.69(d,J=9.2Hz,1H),5.76(dt,J=12.2,6.0Hz,1H),5.28(ddd ,J=12.6,9.1,3.9Hz,1H),4.71(d,J=11.9Hz,1H),4.50(d,J=11.9Hz,1H),4.25(q,J=5.8Hz,2H),3.51-3.44(m,4H),3.31-3.22(m,1H),2.8 1(d,J=4.8Hz,3H),2.77(d,J=4.8Hz,3H),2.57-2.47(m,2H),2.05(q,J=6.3Hz,2H),1.98(td,J=9.4,4.1Hz,1H),1.61(s,3H),1.53(s,3H).

[0453] LCMS: 854 [M+H] + .

[0454] Example 28: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[6,4-c]pyrazol-3-yl)-N 4 -(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo- Synthesis of 2,3-dihydro-1H-benzo[de]isoquinolin-5-yl)oxy)hexyl)terephthalamide (28).

[0455]

[0456] According to the synthetic route of compound 1 and 5-((6-aminohexyl)oxy)-2-(2,6-dioxopiperidin-3-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione in Example 1, compound 28 (6.9 mg) was obtained.

[0457] 1H NMR(500MHz,DMSO-d6)δ10.99(s,1H),10.94(s,1H),8.96(s,1H),8.55(s,1H),8.37-8.20(m,2H),8.06-7.98(m,3H),7.97-7.83(m,4H), 7.77(q,J=8.5Hz,1H),7.41-7.29(m,4H),7.28-7.19(m,1H),6.69(d,J=9.1Hz,1H),5.75(dt,J=11.7,5.9Hz,1H),5.28(ddd,J=12.6,9.1, 3.9Hz,1H),4.71(d,J=11.9Hz,1H),4.50(d,J=11.9Hz,1H),4.16(q,J=6.4Hz,2H),3.52-3.45(m,1H),3.26(d,J=22.8Hz,5H),2.82(d,J= 4.7Hz,3H),2.77(d,J=4.8Hz,3H),2.59-2.47(m,2H),2.03-1.90(m,1H),1.84-1.71(m,2H),1.61(s,3H),1.56-1.42(m,5H),1.37(s,2H).

[0458] LCMS: 896 [M+H] + .

[0459] Example 29: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3- (2,3-dihydro-1H-benzo[de]isoquinolin-5-yl)oxy)ethoxy)ethoxy)ethyl)terephthalamide Synthesis of (29).

[0460]

[0461] According to the synthetic route of compound 1 and 5-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione in Example 1, compound 29 (4.5 mg) was obtained.

[0462] 1H NMR (500MHz, DMSO-d6) δ10.98(s,1H),10.94(s,1H),8.98(s,1H),8.61(t,J=5.6Hz,1H),8.38-8.16(m,2H),8.08-7.86(m,7H),7. 75(dt,J=10.5,7.8Hz,1H),7.45-7.28(m,4H),7.23(t,J=7.2Hz,1H),6.68(d,J=9.2Hz,1H),5.75(dt,J=11.7,5.9Hz,1H),5.28(dd d,J=12.2,9.2,3.9Hz,1H),4.69(d,J=11.8Hz,1H),4.54-4.40(m,1H),4.27(q,J=5.5Hz,2H),3.79(q,J=5.2Hz,2H),3.64-3.43(m ,9H),3.41-3.35(m,2H),2.79(dd,J=22.8,4.7Hz,6H),2.61-2.46(m,2H),1.97(dd,J=11.9,6.2Hz,1H),1.61(s,3H),1.53(s,3H).

[0463] LCMS: 928 [M+H] + .

[0464] Example 30: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4- methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)p-phenylene Synthesis of diformamide (30).

[0465]

[0466] According to the synthetic route of compound 1 and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide in Example 1, compound 30 (7.1 mg) was obtained.

[0467] 1H NMR (500MHz, DMSO-d6) δ12.50(s,1H),11.07(s,1H),8.99(s,1H),8.42(d,J=7.8Hz,1H),8.20-8.14(m,1H),8.10(d,J=8.2Hz,2H),8.00(d,J=8.1H z,2H),7.48-7.43(m,2H),7.42-7.38(m,4H),7.33(t,J=7.6Hz,2H),7.23 (t,J=7.3Hz,1H),6.38(s,1H),5.15(d,J=3.5Hz,1H),5.06-4.90(m,2H),4 .79(d,J=9.1Hz,1H),4.58(t,J=12.1Hz,2H),4.47(t,J=8.1Hz,1H),4.33 (s,1H),3.69(d,J=3.1Hz,2H),2.86(d,J=19.3Hz,6H),2.47(s,3H),2.42- 2.27(m,1H),2.05(ddd,J=12.6,7.6,2.7Hz,1H),1.82(ddd,J=12.9,8.6, 4.5Hz, 1H), 1.66 (s, 3H), 1.59 (s, 3H), 1.39 (d, J = 7.0Hz, 3H), 1.06 (s, 9H).

[0468] LCMS: 917 [M+H] + .

[0469] Example 31: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of 2-amino)-2-oxoethyl)terephthalamide (31).

[0470]

[0471] According to the synthetic route of compound 1 in Example 1 and (2S,4R)-1-((S)-2-(2-aminoacetamido)-3,3-dimethylbutyryl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, compound 31 was obtained.

[0472] LCMS: 974 [M+H] + .

[0473] Example 32: 8-(4-((5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl- 1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzoyl)-N-((S)-1-((2S,4R)-4- Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethoxy Synthesis of 2-(4-(2-(4-(2-(4-(2-nitro-1-yl)-1-oxobutan-2-yl)-8-azaspiro[4.5]decane-2-carboxamide (32).

[0474]

[0475] According to the synthetic route of compound 1 in Example 1 and N-((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)-8-azaspiro[4.5]decane-2-carboxamide, compound 32 (11.3 mg) was obtained.

[0476] 1 H NMR (500MHz, DMSO-d6) δ12.51(s,1H),11.02(s,1H),8.99(s,1H),8.39(d,J=7.8Hz,1H),8.07(d,J=7.9Hz,2H),7.74(s,1H),7.51( d,J=7.8Hz,2H),7.47-7.31(m,9H),7.25(t,J=7.3Hz,1H),6.48(s,1H),5.12(s,2H),4.92(p,J=7.1Hz,1H),4.65(s,1H),4.54(dd,J =24.8,10.4Hz,2H),4.43(t,J=8.1Hz,1H),4.29(s,1H),3.60(s,4H),3.29(s,2H),2.99(d,J=14.4Hz,6H),2.46(s,3H),2.02(t,J=1 0.4Hz,1H),1.84-1.75(m,4H),1.67(s,3H),1.59(s,3H),1.55-1.44(m,8H),1.38(d,J=7.0Hz,3H),0.94(s,9H),0.89-0.79(m,1H).

[0477] LCMS: 1082 [M+H] + .

[0478] Example 33: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of (4-(2-carbamoyl)phenyl)terephthalamide (33).

[0479]

[0480] (S)-3-(4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzamido)benzoic acid

[0481] To a solution of (S)-4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzoic acid (15 mg, 0.025 mmol) and DIEA (12.9 mg, 0.1 mmol) in dry DCM (1 mL) was added propionic anhydride (50% wt in EA, 32 μL, 0.05 mmol). The reaction was stirred for 10 minutes before the addition of tert-butyl 3-aminobenzoate (4.8 mg, 0.025 mmol). The mixture was stirred for 30 minutes and then concentrated in vacuo. The crude product was dissolved in DCM (1 mL) before the addition of TFA (0.5 mL). After 1 hour, the solvent was removed in vacuo to give the crude product without further purification.

[0482] LCMS: 666 [M+H]+.

[0483]

[0484] To a solution of (S)-3-(4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzamido)benzoic acid (15 mg, 0.025 mmol) and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazolyl-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride (13 mg, 0.025 mmol) and DIEA (13 mg, 0.098 mmol) in DMF (1 mL) was added HATU (18 mg, 0.049 mmol). The mixture was stirred at room temperature for 10 minutes and then purified by reverse phase HPLC to give compound 33 as a TFA salt (1.8 mg, 6%).

[0485] LCMS: 1036 [M+H] + .

[0486] Example 34: N 1 -(5-(((S)-2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1, 4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-N 4 -(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) Synthesis of (4-(2-(4 ...

[0487]

[0488] (S)-3-(4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzamido)bicyclo[1.1.1]pentane-1-carboxylic acid

[0489] To a solution of (S)-4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzoic acid (20 mg, 0.040 mmol) and DIEA (21 mg, 0.16 mmol) in dry DCM (1 mL) was added propionic anhydride (50% wt in EA, 52 μL, 0.08 mmol). The reaction was stirred for 10 minutes before the addition of methyl 3-aminobicyclo[1.1.1]pentane-1-carboxylate (8.4 mg, 0.060 mmol). The mixture was stirred for 30 minutes and then concentrated in vacuo. The crude product was dissolved in THF (1 mL) before the addition of LiOH (0.5 mL, 1 M aq.). After 1 hour, the reaction was acidified with aqueous hydrochloric acid. The mixture was extracted with EA. The organic layer was collected and concentrated in vacuo. The resulting residue was purified by reverse phase HPLC to give the title compound (29.8 mg, 0.04 mmol).

[0490] LCMS: 600[M+H] + .

[0491]

[0492] To a solution of (S)-3-(4-((5-((2-(dimethylamino)-1-phenylethyl)carbamoyl)-6,6-dimethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)carbamoyl)benzamido)bicyclo[1.1.1]pentane-1-carboxylic acid (29 mg, 0.04 mmol) and DIEA (31 mg, 0.24 mmol) in dry DCM (1 mL) was added propionic anhydride (50% wt in EA, 62 μL, 0.096 mmol). The reaction was stirred for 10 minutes before the addition of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride (25 mg, 0.048 mmol). The mixture was stirred for 30 minutes and then concentrated in vacuo. The residue was purified by reverse phase HPLC to give compound 34 as a TFA salt (11 mg, 0.0096 mmol, 24%).

[0493] 1 H NMR(500MHz,DMSO-d6)δ12.57(s,1H),11.09(s,1H),9.30(s,1H),9.23(s,1H),8 .99(s,1H),8.43(dd,J=7.8,1.7Hz,1H),8.10(dd,J=8.5,6.1Hz,2H),7.99(t,J= 10.9Hz,2H),7.45(dq,J=8.3,1.7Hz,4H),7.42-7.37(m,4H),7.33-7.26(m,2H), 6.77(d,J=8.9Hz,1H),5.37(d,J=11.2Hz,1H),5.17(d,J=3.5Hz,1H),4.97-4.89( m,1H),4.80(dd,J=11.2,7.9Hz,1H),4.62-4.54(m,2H),4.46(q,J=8.5Hz,1H),4 .30(s,1H),3.69(s,1H),3.65-3.52(m,2H),2.86(d,J=19.5Hz,6H),2.46(s,3H), 2.33(s,4H),2.10-2.01(m,1H),1.86-1.76(m,1H),1.69(s,3H),1.60(s,3H),1. 39(dd,J=7.0,1.7Hz,3H),1.32-1.23(m,1H),1.19(t,J=7.3Hz,1H),0.96(s,9H).

[0494] LCMS: 1026 [M+H] + .

[0495] Example 35: Degradation of CDK7 using bispecific compounds 1-26 of the invention.

[0496] Jurkat cells were treated with DMSO or 1 μM of compound 1-26 for 6 h. Then, the cells were incubated in radioimmunoprecipitation assay (RIPA) buffer ( Cells were lysed by Pierce Biosciences. Protein concentration was determined by bicinchoninic acid assay (BCA) analysis (Pierce TM ). Equal amounts of protein were measured by 4-12% Tris-Base gel (Invitrogen TM ) were analyzed and then transferred to polyvinylidene difluoride (PVDF) membranes (BioRad) for immunoblotting and stained with primary antibodies against CDK7 (cell signaling) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) Immunoblotting was performed and then 800 labeled goat anti-rabbit immunoglobulin G (IgG) and Goat anti-mouse IgG labeled with 680 The membranes were immunoblotted with secondary antibodies. Detected on CLx system.

[0497] Figure 1A The results shown indicate that bispecific compounds 1, 3, 6, 7, 8, 9 and 10 induced degradation of CDK7 after 6 hours.

[0498] Figure 2A The results shown indicate that bispecific compounds 3, 11, 12, 13, 15, 16 and 20 induced degradation of CDK7 after 6 hours.

[0499] Figure 1C The results shown indicate that bispecific compounds 21, 22, 23, 25 and 26 induced the degradation of CDK7 after 6 hours.

[0500] Example 36: CDK7 degradation is ligand- and proteasome-dependent.

[0501] The cells were treated with 10 μM YKL-5-124 (parent compound and known CDK7 inhibitor), 10 μM DGY-05-180 (VHL ligand), 0.2 μM bortezomib (a proteasome inhibitor available, for example, from Millipore Sigma, Cat. No. 179324-69-7, Burlington, MA), and 1 μM MLN4924 (a ubiquitin inhibitor available, for example, from MedChemExpress). Jurkat cells were pretreated with 1 μM of compound 3 or 20 for 2 hours and then treated for 4 hours with 1 μM of compound 3 or 20. Cells were lysed and immunoblotted with CDK7 and GAPDH antibodies as described in Example 27. The structures of YKL-5-124, DGY-05-180, bortezomib, and MLN4924 are as follows.

[0502]

[0503] YKL-5-124

[0504] DGY-05-180

[0505]

[0506] Bortezomib MLN4924 (Pevonedistat)

[0507] The results are as follows Figure 2A and 2B They showed that YKL-5-124, DGY-05-180, bortezomib, and MLN4924 rescued CDK7 degradation induced by bispecific compounds 3 and 20. The results suggest that CDK7 degradation is ligand- and proteasome-dependent.

[0508] All patent publications and non-patent publications are indicative of the level of skill of those skilled in the art to which the invention pertains. All of these publications are incorporated herein by reference as if each individual publication was specifically and individually indicated to be incorporated herein by reference.

[0509] Although the present invention has been described herein with reference to specific embodiments, it should be understood that these embodiments are merely illustrative of the principles and applications of the present invention. Therefore, it should be understood that many modifications may be made to the illustrative embodiments and other configurations may be designed without departing from the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A bispecific compound comprising a portion and a degradation determinant Degron (D) covalently linked to each other via a linker Linker (L), wherein the portion binds to cyclin-dependent kinase 7 (CDK7), wherein the compound has a structure represented by formula (I): or a pharmaceutically acceptable salt or stereoisomer thereof, wherein R1 stands for in R 1” It is hydrogen, R 1a is phenyl, and R 2a Yes – NR 1N R 2N , where R 1N and R 2N Each is independently a C1-C6 alkyl group; R3 and R4 each independently represent a C1-C6 alkyl group; R5 represents hydrogen; L1 stands for -NR L1 -、-NR L1 C(=O)-, -C(=O)NR L1 -, where R L1 It is hydrogen; A represents phenyl; L2 represents a bond, -C(=O)-, -C(=O)NR L2 -、-NR L2 C(=O)-, where R L2 It is hydrogen; B represents a bond; and R2 does not exist, The linker is an alkylene chain or a polyethylene glycol chain, any of which may be interrupted and / or terminated at either or both ends by at least one of the following: –O–, –S–, –N(R′)–, –C≡C–, –C(O)–, –C(O)O–, –OC(O)–, –OC(O)O–, –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’)–, –S(O)2–, –OS(O)–, –S(O)O–, –S(O)–, –OS(O)2–, –S(O)2 O–, –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-C 12 carbocyclylene, 3- to 12-membered heterocyclylene, 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or C1-C6 alkyl, wherein the interrupting group and one or two terminating groups may be the same or different, wherein the alkylene chain contains 1-12 methylene units, and the polyethylene glycol chain contains 2-10 ethylene glycol units; and wherein the degron is represented by any one of structures (D1-a) to (D1-h) and (D2-a), (D2-b) and (D2e):

2. The bispecific compound of claim 1, wherein R1 is 3. The bispecific compound of claim 1, wherein R3 and R4 are both methyl.

4. The bispecific compound of claim 1, wherein L1 is -NHC(O)-.

5. The bispecific compound of claim 1, wherein L2 is a bond or -NHC(O)-.

6. The bispecific compound of claim 1, wherein L1 is -NHC(O)-, R3 and R4 are methyl, and R5 is H, and has a structure represented by (I-1b): or a pharmaceutically acceptable salt or stereoisomer thereof.

7. The bispecific compound according to claim 1, wherein R3 and R4 are methyl groups, and R5 is H, and has a structure represented by formula (I-2c) or (I-2d): or a pharmaceutically acceptable salt or stereoisomer thereof.

8. The bispecific compound of claim 1, wherein L1 is -NHC(O)-, R1 is R3 and R4 are methyl groups, and R5 is H, and it has a structure represented by formula (I-3c): or a pharmaceutically acceptable salt or stereoisomer thereof.

9. The bispecific compound of claim 1, wherein R1 is R3 and R4 are methyl groups, and R5 is H, which has a structure represented by any one of formula (I-4e) or (I-4g): or a pharmaceutically acceptable salt or stereoisomer thereof.

10. The bispecific compound of claim 1, wherein L1 is -NHC(O)-, R1 is R3 and R4 are methyl groups, and R5 is H, which has a structure represented by formula (I-5c): or a pharmaceutically acceptable salt or stereoisomer thereof.

11. The bispecific compound of claim 1, wherein R1 is R3 and R4 are methyl groups, and R5 is H, and has a structure represented by any one of formula (I-6e) or (I-6g): or a pharmaceutically acceptable salt or stereoisomer thereof.

12. The bispecific compound according to claim 1, which has a structure represented by formula (I-58): or a pharmaceutically acceptable salt or stereoisomer thereof.

13. The bispecific compound of claim 1, wherein the linker is represented by any one of structures (L11) to (L28):

14. The bispecific compound according to claim 1, which is represented by any one of formulas (I-59) to (I-71): or a pharmaceutically acceptable salt or stereoisomer thereof.

15. The bispecific compound of claim 1, wherein the linker is an alkylene chain which may be interrupted and / or terminated at either or both ends by at least one of: –O–, –S–, –N(R’)–, –C≡C–, –C(O)–, –C(O)O–, –OC(O)–, –OC(O)O–, –C(O)N(R’)–, –N(R’)C(O)–, C3-C 12 Carbocyclylene, 3- to 12-membered heterocyclylene, 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or C1-C6 alkyl, wherein the interrupting group and one or two terminating groups may be the same or different.

16. The bispecific compound of claim 1, wherein the linker is a polyethylene glycol chain that may terminate at either or both termini at at least one of: -S-, -N(R')-, -C≡C-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R')-, -N(R')C(O)-, C3-C 12 Carbocyclylene, 3- to 12-membered heterocyclylene, 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or C1-C6 alkyl, wherein one or both terminating groups may be the same or different.

17. The bispecific compound of claim 1, wherein the degron is represented by any one of structures (D1-a) to (D1-h):

18. The bispecific compound of claim 1, which is represented by any one of Formulas (I-72a) to (I-72h): or a pharmaceutically acceptable salt or stereoisomer thereof.

19. The bispecific compound of claim 1, wherein the degron is represented by any one of structures (D2-a), (D2-b), and (D2-e):

20. The bispecific compound of claim 1, which is represented by any one of Formula (I-73a), (I-73b) and (I-73e): or a pharmaceutically acceptable salt or stereoisomer thereof.

21. The bispecific compound of claim 1, which is represented by any one of formulas (1) to (34): or pharmaceutically acceptable salts and stereoisomers thereof.

22. A pharmaceutical composition comprising a therapeutically effective amount of the bispecific compound of any one of claims 1-21 or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

23. Use of the bispecific compound of any one of claims 1-21, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating a disease or condition mediated by aberrant CDK7 activity.

24. The use according to claim 23, wherein the disease is cancer.

25. The use according to claim 24, wherein the cancer is a solid tumor.

26. The use according to claim 25, wherein the solid tumor is breast cancer, brain cancer, lung cancer, colorectal cancer, neuroblastoma or osteosarcoma.

27. The use according to claim 24, wherein the cancer is a hematological cancer.

28. The use according to claim 27, wherein the hematological cancer is leukemia, lymphoma or multiple myeloma.

29. The use according to claim 23, wherein the disease is an autoimmune disease.

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