Therapeutic combinations including ubiquitin-specific processing protease 1 (USP1) inhibitors and PARP1-specific inhibitors

Abstract

The present disclosure provides a therapeutic combination comprising (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor and (ii) a PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, wherein the USP1 inhibitor is: The present disclosure also provides a therapeutic combination comprising TIFF2025533524000028.tif139111 or its pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal.The present disclosure also relates to the use of said combination for inhibiting USP1 and / or PARP1 protein and / or treating disorders that respond to the inhibition of USP1 and / or PAR1P protein and USP1 and / or PARP1 activity.The combination of the present disclosure is particularly useful for treating cancer.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 376,987, filed September 23, 2022, U.S. Provisional Patent Application No. 63 / 380,742, filed October 24, 2022, and U.S. Provisional Patent Application No. 63 / 496,289, filed April 14, 2023, each of which is incorporated herein by reference in its entirety.

[0002] Reference to an electronically submitted sequence listing The contents of the electronically submitted sequence listing (Name: 4195_034PC03_Seqlisting_ST26; Size: 2,057 bytes; and Created: September 21, 2023) are incorporated herein by reference in their entirety.

[0003] Field The present disclosure provides a therapeutic combination of a ubiquitin-specific processing protease 1 (USP1) inhibitor and a selective poly(ADP-ribose) polymerase (PARP1) inhibitor. A method for treating cancer is provided, comprising administering the combination. [Background technology]

[0004] Ubiquitin is a small (76 amino acid) protein that binds to target proteins after transcription. The outcome of ubiquitination is determined by the number and linkage topology of ubiquitin molecules attached to the target protein. For example, proteins displaying lysine 48-linked polyubiquitin chains are generally targeted to the proteasome for degradation, whereas monoubiquitination or polyubiquitin chains attached via other lysines regulate non-proteolytic functions such as cell cycle regulation, DNA damage repair, transcription, and endocytosis. Ubiquitination is a reversible process; enzymes called deubiquitinases remove ubiquitin from target proteins.

[0005] USP1 is a deubiquitinating enzyme involved in DNA damage repair. USP1 interacts with UAF1 (USP1-associated factor 1) to form a complex required for deubiquitinase activity. The USP1 / UAF1 complex deubiquitinates monoubiquitinated PCNA (proliferating cell nuclear antigen) and monoubiquitinated FANCD2 (Fanconi anemia complementation group D2), proteins that play important functions in the translesion synthesis (TLS) and Fanconi anemia (FA) pathways, respectively. The USP1 / UAF1 complex also deubiquitinates Fanconi anemia complementation group I (FANCI). These two pathways are essential for repair of DNA damage induced by DNA crosslinking agents such as cisplatin and mitomycin C (MMC).

[0006] The poly(ADP-ribose) polymerase (PARP) family of enzymes is involved in DNA repair and genome integrity. PARP is important in the single-strand break repair pathway and base excision repair pathway. Its key enzymatic activity is the addition of ADP-ribose to substrate proteins via the cleavage of NAD+ and the liberation of nicotinamide. This poly(ADP-ribosyl)ation ("PARylation") activity is activated by DNA strand breaks, allowing PARP to add Par to itself and other DNA repair enzymes. PARP is important for recruiting DNA repair proteins to the site of damage.

[0007] Homologous recombination is a DNA repair process essential for the accurate repair of DNA damage. The BRCA1 / 2 genes, along with other Fanconi anemia pathway genes (e.g., RAD51D, NBN, and ATM), are components of homologous recombination-mediated DNA repair. Mutations in genes encoding homologous recombination factors are involved in the development of certain cancers. PARP inhibitors prevent the repair of DNA single-strand breaks and promote the conversion of single-strand breaks to double-strand breaks, resulting in synthetic lethality in cancer cells that lack double-strand break mechanisms such as homologous recombination.

[0008] There remains an unmet medical need for more effective therapies, including combination therapies, for the treatment of cancer. Summary of the Invention

[0009] Provided herein is a combination of (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal thereof, and (ii) a selective poly ADP-ribose polymerase 1 inhibitor (selective PARP1 inhibitor), or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal thereof. Also provided herein is a method of treating a subject with cancer using such a combination.

[0010] In one aspect, the disclosure provides a combination composition comprising (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor and (ii) a PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, wherein the USP1 inhibitor is: (a) Formula I: [ka] (b) Formula II: [ka] (c) Formula III: [ka] The present invention relates to a composition that is a compound selected from the group consisting of: and a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In another embodiment, Formula I, Formula II, or Formula III is provided as a co-crystal. In some embodiments, Formula I, Formula II, or Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0011] In some embodiments, the PARP1 selective inhibitor is selected from the group consisting of AZD5305, and a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

[0012] In one embodiment, the USP1 inhibitor is a compound of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula I is provided as a co-crystal with a pharmaceutically acceptable acid.

[0013] In another embodiment, the USP1 inhibitor is a compound of Formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula II is provided as a co-crystal with a pharmaceutically acceptable acid.

[0014] In another embodiment, the USP1 inhibitor is a compound of Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0015] In another embodiment, the USP1 inhibitor is a co-crystal of a compound of Formula I and a second pharmaceutically acceptable compound. In some embodiments, the pharmaceutically acceptable co-crystal is formed between a compound of Formula I and a pharmaceutically acceptable acid. In some embodiments, the pharmaceutically acceptable acid is selected from the group consisting of benzoic acid, gentisic acid, and salicylic acid.

[0016] In another embodiment, the USP1 inhibitor is a co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0017] In another embodiment, the USP1 inhibitor is a co-crystal of gentisic acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0018] In another embodiment, the USP1 inhibitor is the gentisate salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0019] In one aspect, the present disclosure relates to a combination composition for the manufacture of a medicament for treating cancer.

[0020] In another aspect, the present disclosure relates to a pharmaceutical combination composition comprising a combination composition and a pharmaceutically acceptable carrier.

[0021] In one embodiment, the pharmaceutical composition is for the treatment of cancer.

[0022] In one aspect, the present disclosure relates to a kit comprising a combination composition or pharmaceutical combination composition and instructions for administering the combination to a subject with cancer.

[0023] In another aspect, the present disclosure provides a method of treating cancer in a subject, comprising administering to the subject (i) a USP1 inhibitor and (ii) a PARP1 selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, USP1 inhibitors (a) Formula I: [ka] (b) Formula II: [ka] (c) Formula III: [ka] and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof, In another embodiment, Formula I, Formula II, or Formula III is provided as a co-crystal. In some embodiments, Formula I, Formula II, or Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0024] In one embodiment of the method, the PARP1 selective inhibitor is AZD5305 or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof.

[0025] In one embodiment of the method, the USP1 inhibitor is a compound of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula I is provided as a co-crystal with a pharmaceutically acceptable acid.

[0026] In another embodiment of the method, the USP1 inhibitor is a compound of Formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula II is provided as a co-crystal with a pharmaceutically acceptable acid.

[0027] In another embodiment of the method, the USP1 inhibitor is a compound of Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0028] In another embodiment, the USP1 inhibitor is a co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0029] In another embodiment, the USP1 inhibitor is a co-crystal of gentisic acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0030] In another embodiment, the USP1 inhibitor is the gentisate salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0031] In one embodiment of the present disclosure, administration of a USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof and a selective PARP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof provides a synergistic effect.

[0032] In one embodiment of the present disclosure, the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, and the PARP1-selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof are administered in a therapeutically effective amount sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) a reduction in tumor size; (ii) an increase in the rate of cancer tumor regression; (iii) a reduction or inhibition of cancer tumor growth; and (iv) a reduction in the toxicity of the PARP1-selective inhibitor administered as monotherapy (e.g., due to a lower amount of the PARP1-selective inhibitor being administered). In one embodiment of the present disclosure, the USP1 inhibitor or its pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal, and the PARP1-selective inhibitor or its pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal, are administered in a therapeutically effective amount sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) a reduction in tumor size, (ii) an increase in cancer tumor regression rate, and (iii) a reduction or inhibition of cancer tumor growth. In one embodiment of the present disclosure, the USP1 inhibitor or its pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal, and the PARP1-selective inhibitor or its pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal, are administered in an amount sufficient to reduce the toxicity of the PARP1-selective inhibitor administered as monotherapy (e.g., due to the administration of a lower amount of the PARP1-selective inhibitor).

[0033] In one embodiment, the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof, and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof are administered in a therapeutically effective amount sufficient to delay, reduce, or prevent tumor rebound (rapid regrowth).

[0034] In one embodiment, the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof are administered sequentially.

[0035] In another embodiment, the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof are administered simultaneously.

[0036] In one aspect of the disclosure, the combination is administered to a mammal, hi another aspect, the mammal is a human.

[0037] In some embodiments, the cancer is selected from the group consisting of a hematological cancer, a lymphatic cancer, a solid tumor, a DNA damage repair pathway deficient cancer, and a homologous recombination deficient cancer.

[0038] In some embodiments, the cancer is an advanced solid tumor.

[0039] In some embodiments, the cancer is selected from the group consisting of brain cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, skin cancer, uterine cancer, peritoneal cancer, and endometrial cancer, and breast cancer.

[0040] In some embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0041] In some embodiments, the cancer is colon cancer.

[0042] In some embodiments, the cancer is bladder cancer.

[0043] In some embodiments, the cancer is ovarian cancer or breast cancer.

[0044] In some embodiments, the cancer is ovarian cancer.

[0045] In some embodiments, the cancer is breast cancer.

[0046] In some embodiments, the cancer is triple-negative breast cancer.

[0047] In some embodiments, the cancer is selected from the group consisting of bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma, and meningioma; soft tissue cancer, including rhabdoid and sarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer, uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.

[0048] In some embodiments, the cancer is a DNA damage repair pathway deficient cancer.

[0049] In some embodiments, the cancer is a BRCA1 mutation cancer. In some embodiments, the BRCA1 mutation is a germline mutation. In some embodiments, the BRCA1 mutation is a somatic mutation. In some embodiments, the BRCA1 mutation causes a BRCA1 deficiency.

[0050] In some embodiments, the cancer is a BRCA2-mutated cancer. In some embodiments, the BRCA2 mutation is a germline mutation. In some embodiments, the BRCA2 mutation is a somatic mutation. In some embodiments, the BRCA2 mutation causes a BRCA2 deficiency.

[0051] In some embodiments, the cancer is a BRCA1 mutant cancer and a BRCA2 mutant cancer.

[0052] In some embodiments, the cancer is a BRCA1-deficient cancer.

[0053] In some embodiments, the cancer is a BRCA2-deficient cancer.

[0054] In some embodiments, the cancer is a BRCA1-deficient cancer and a BRCA2-deficient cancer.

[0055] In some embodiments, cancer is PARP inhibitor refractory or resistant cancer.In some embodiments, cancer is PARP inhibitor resistant or refractory BRCA1 mutation cancer, BRCA2 mutation cancer or BRCA1 and BRCA2 mutation cancer.In some embodiments, cancer is PARP inhibitor resistant or refractory BRCA1 deficient cancer, BRCA2 deficient cancer or BRCA1 and BRCA2 deficient cancer.

[0056] In some embodiments, cancer is olaparib-refractory or resistant cancer.In some embodiments, cancer is olaparib-resistant or refractory BRCA1 mutation cancer, BRCA2 mutation cancer, or BRCA1 and BRCA2 mutation cancer.In some embodiments, cancer is olaparib-resistant or refractory BRCA1-deficient cancer, BRCA2-deficient cancer, or BRCA1 and BRCA2-deficient cancer.

[0057] In some embodiments, the cancer is AZD-5305 refractory or resistant cancer. In some embodiments, the cancer is AZD-5305 resistant or refractory BRCA1 mutant cancer, BRCA2 mutant cancer, or BRCA1 and BRCA2 mutant cancer. In some embodiments, the cancer is AZD-5305 resistant or refractory BRCA1 deficient cancer, BRCA2 deficient cancer, or BRCA1 and BRCA2 deficient cancer.

[0058] In some embodiments, the cancer has a mutation in the gene encoding the ataxia telangiectasia mutated (ATM) protein kinase. In some embodiments, the ATM mutation is a germline mutation. In some embodiments, the ATM mutation is a somatic mutation. In some embodiments, the cancer is an ATM-deficient cancer.

[0059] In some embodiments, the cancer comprises cancer cells having a mutation in the gene encoding p53. In some embodiments, the mutation in the gene encoding p53 is a germline mutation. In some embodiments, the mutation in the gene encoding p53 is a somatic mutation. In some embodiments, the cancer comprises cancer cells having a loss-of-function mutation in the gene encoding p53.

[0060] In some embodiments, the cancer has a mutation in a gene encoding at least two of p53, BRCA1, BRCA2, and ATM.

[0061] In some embodiments, the cancer has been previously treated with platinum therapy. In some embodiments, the cancer is platinum-resistant or platinum-refractory. In some embodiments, the platinum therapy is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, and combinations thereof.

[0062] In another aspect, the present disclosure relates to a method for treating a USP1 protein-mediated disorder and / or a PARP protein-mediated disorder, comprising administering to a subject in need of said treatment a USP1 inhibitor of Formula I, Formula II or Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof, and a PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof, in an amount effective to treat the USP1 protein-mediated disorder and / or the PARP protein-mediated disorder.

[0063] In another aspect, the present disclosure relates to a method of inhibiting USP1 protein and / or PARP protein, comprising contacting the USP1 protein and / or PARP protein with a USP1 inhibitor of Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, and a PARP1 selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

[0064] In some embodiments, the contacting occurs in vitro.

[0065] In some embodiments, the contacting occurs in vivo.

[0066] In one embodiment of the method, the PARP1 selective inhibitor is AZD5305 or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof.

[0067] In one embodiment of the method, the USP1 inhibitor is a compound of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula I is provided as a co-crystal with a pharmaceutically acceptable acid.

[0068] In another embodiment of the method, the USP1 inhibitor is a compound of Formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula II is provided as a co-crystal with a pharmaceutically acceptable acid.

[0069] In another embodiment of the method, the USP1 inhibitor is a compound of Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0070] In another embodiment, the USP1 inhibitor is a co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0071] In another embodiment, the USP1 inhibitor is a co-crystal of gentisic acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0072] In another embodiment, the USP1 inhibitor is the gentisate salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0073] In one aspect, the disclosure relates to the use of the combination for the manufacture of a medicament for the treatment of cancer.

[0074] In another aspect, the present disclosure relates to a pharmaceutical composition comprising the combination and a pharmaceutically acceptable carrier.

[0075] In one embodiment, the pharmaceutical composition is for use in the treatment of cancer.

[0076] In one aspect, the present disclosure relates to a kit comprising a combination or pharmaceutical composition and instructions for administering the combination to a subject with cancer.

[0077] Additional aspects and advantages of the disclosure will be set forth in part in the description which follows, and in part may be derived from the description, or may be learned by practice of the disclosure. The aspects and advantages of the disclosure will be realized and attained by means of the elements and combinations particularly pointed out in the claims.

[0078] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. [Brief explanation of the drawings]

[0079] [Figure 1A] FIG. 1A shows that the USP1 inhibitor of formula I enhances the antitumor activity of AZD5305 in a breast cancer PDX model.

[0080] [Figure 1B] FIG. 1B shows the efficacy of a combination of a USP1 inhibitor of formula I and AZD5305 in mice bearing tumors resistant to PARP inhibitors.

[0081] [Figure 1C] FIG. 1C shows the tolerability of the combination of a USP1 inhibitor of formula I with AZD5305.

[0082] [Figure 2] FIG. 2 shows the XRPD pattern corresponding to crystalline form 2.

[0083] [Figure 3] FIG. 3 shows the DSC and TGA thermograms corresponding to crystalline Form 2.

[0084] [Figure 4] FIG. 4 shows colony formation assays (CFU) of UWB1.289 parental cells treated with Formula I and / or AZD5305.

[0085] [Figure 5] FIG. 5 shows colony formation assays (CFU) of UWB1.289BRCA1-overexpressing cells treated with Formula I and / or AZD5305. DETAILED DESCRIPTION OF THE INVENTION

[0086] Detailed Description of Disclosure One aspect of the present disclosure is based on the use of a combination of a ubiquitin-specific processing protease 1 (USP1) protein inhibitor and a PARP1-selective inhibitor, which is useful for inhibiting USP1 and / or PARP proteins and for treating diseases, disorders, or conditions that respond to the inhibition of USP1 and / or PARP proteins, such as cancer.

[0087] In some embodiments, the combination of a USP1 inhibitor and a PARP1 selective inhibitor provides a synergistic effect.

[0088] In some embodiments, the USP1 inhibitor and PARP1 selective inhibitor are administered in a therapeutically effective amount sufficient to produce a therapeutic effect including (i) a reduction in tumor size, (ii) an increase in the rate of cancer tumor regression, (iii) a reduction or inhibition of cancer tumor growth, and / or (iv) a reduction in the toxicity of the PARP1 selective inhibitor administered as monotherapy (e.g., due to a lower amount of the PARP1 selective inhibitor administered). In some embodiments, the USP1 inhibitor and PARP1 selective inhibitor can delay, reduce, or prevent tumor rebound (rapid regrowth).

[0089] The tolerability (lack of toxicity) of the combinations provided herein is particularly surprising given that other combinations with the PARP inhibitor olaparib have not been well tolerated. See, e.g., Samol, J., et al., Invest. New Drugs, 30:1493-500 (2012) (“Further development of olaparib and topotecan in combination was not explored due to dose-limiting hematological AEs and the resulting sub-therapeutic MTD.”).

[0090] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In case of conflict, the present application, including definitions, will control. Unless otherwise required by context, singular terms shall include plurals and plural terms shall include the singular. All publications, patents, and other references mentioned herein are incorporated by reference in their entirety for all purposes as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference herein.

[0091] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and are not intended to be limiting. Other features and advantages of the present disclosure will be apparent from the detailed description and claims.

[0092] In order to further define this disclosure, the following terms and definitions are provided.

[0093] Embodiments described herein are understood to include embodiments "consisting of" and / or "consisting essentially of." As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated. The use of the term "or" herein does not imply that alternatives are mutually exclusive.

[0094] In this application, the use of "or" means "and / or" unless expressly stated or understood by one of ordinary skill in the art. In the context of multiple dependent claims, the use of "or" refers back to the preceding multiple independent or dependent claims.

[0095] The term "and / or" as used herein is considered as a specific disclosure of two specific features or components, each with or without the other. Thus, the term "and / or" used in phrases such as "A and / or B" herein is intended to include "A and B," "A or B," "A" (single), and "B" (single). Similarly, the term "and / or" used in phrases such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B, or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0096] As used herein, the term "about" includes the stated number ±10%. Thus, "about 10" means 9 to 11. As will be understood by those skilled in the art, reference to a value or parameter herein as "about" includes (describes) an example directed to that value or parameter itself. For example, a description referring to "about X" includes a description of "X."

[0097] The present disclosure encompasses the preparation and use of salts of USP1 inhibitors and PARP1 selective inhibitors, including non-toxic pharmaceutically acceptable salts. Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts and basic salts. Pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium salts, potassium salts, cesium salts, and the like; alkaline earth metal salts such as calcium salts and magnesium salts; organic amine salts such as triethylamine salts, pyridine salts, picoline salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, and the like; inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulfate, and the like; organic acid salts such as citrate, lactate, tartrate, maleate, fumarate, mandelate, acetate, dichloroacetate, trifluoroacetate, oxalate, formate, and the like; sulfonate salts such as methanesulfonate, benzenesulfonate, p-toluenesulfonate, and the like; and amino acid salts such as alginate, aspartate, glutamate, and the like. As used herein, the term "pharmaceutically acceptable salt" refers to any salt obtained, for example, by reaction of a USP1 inhibitor or PARP1 selective inhibitor of the present disclosure with an acid or base, that is physiologically tolerated in a target patient (e.g., a mammal, e.g., a human).

[0098] Acid addition salts can be formed by mixing a solution of the particular USP1 inhibitor or PARP1 selective inhibitor with a solution of a pharmaceutically acceptable non-toxic acid, such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, dichloroacetic acid, etc. Base salts can be formed by mixing a solution of the disclosed USP1 inhibitor or PARP1 selective inhibitor with a solution of a pharmaceutically acceptable non-toxic base, such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, etc.

[0099] In some embodiments of the present disclosure, a pharmaceutically acceptable salt is formed between a compound of Formula I, Formula II, or Formula III and a pharmaceutically acceptable acid. In some embodiments, the pharmaceutically acceptable salt of a compound of Formula I, Formula II, or Formula III is in a crystalline form.

[0100] In some embodiments, the pharmaceutically acceptable acid is 1-hydroxy-2-naphthoic acid, 4-aminosalicylic acid, ascorbic acid, adipic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, trans-cinnamic acid, citric acid, ethanedisulfonic acid, fumaric acid, galactaric acid, gentisic acid, gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycolic acid, hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid, lactic acid, maleic acid, L-malic acid, malonic acid, R-mandelic acid, meta The carboxylic acid may be selected from the group consisting of toluenesulfonic acid, mucic acid, naphthalenesulfonic acid, nicotinic acid, oxalic acid, palmitic acid, p-toluenesulfonic acid, phosphoric acid, propionic acid, saccharin, salicylic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, vanillic acid, vanillin, ethyl maltol, gallic acid, gallic acid ethyl ester, 4-hydroxybenzoic acid, 4-hydroxybenzoic acid methyl ester, 3,4,5-trihydroxybenzoic acid, nicotinamide, L-proline, and D-sorbitol. In some embodiments, the pharmaceutically acceptable acid is 1-hydroxy-2-naphthoic acid, 4-aminosalicylic acid, ascorbic acid, adipic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, trans-cinnamic acid, citric acid, ethanedisulfonic acid, fumaric acid, galactaric acid, gentisic acid, gallic acid, gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycolic acid, hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid, lactic acid, maleic acid, L-malic acid, malonic acid, R-mandelic acid, In some embodiments, the pharmaceutically acceptable acid is selected from the group consisting of methanesulfonic acid, mucic acid, naphthalenesulfonic acid, nicotinic acid, oxalic acid, palmitic acid, p-toluenesulfonic acid, phosphoric acid, propionic acid, saccharin, salicylic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, vanillic acid, vanillin, ethyl maltol, gallic acid, gallic acid ethyl ester, 4-hydroxybenzoic acid, 4-hydroxybenzoic acid methyl ester, 3,4,5-trihydroxybenzoic acid, nicotinamide, L-proline, and D-sorbitol. In some embodiments, the pharmaceutically acceptable acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, ethanedisulfonic acid, methanesulfonic acid, gentisic acid, benzoic acid, salicylic acid, and gallic acid.In some embodiments, the pharmaceutically acceptable acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, ethanedisulfonic acid, methanesulfonic acid, and gentisic acid. In some embodiments, the pharmaceutically acceptable acid is a benzoic acid derivative. In some embodiments, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with one or more hydroxy groups. In some embodiments, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with one hydroxy group. In some embodiments, the benzoic acid derivative substituted with one hydroxy group is salicylic acid. In some embodiments, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with two hydroxy groups. In some embodiments, the benzoic acid derivative substituted with two hydroxy groups is gentisic acid. In some embodiments, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with three hydroxy groups. In some embodiments, the benzoic acid derivative substituted with three hydroxy groups is gallic acid. In some embodiments, the benzoic acid derivative is selected from the group consisting of salicylic acid, gentisic acid, and gallic acid. In some embodiments, the pharmaceutically acceptable acid is hydrochloric acid. In some embodiments, the pharmaceutically acceptable acid is hydrobromic acid. In some embodiments, the pharmaceutically acceptable acid is ethanedisulfonic acid. In some embodiments, the pharmaceutically acceptable acid is methanesulfonic acid. In some embodiments, the pharmaceutically acceptable acid is gentisic acid. In some embodiments, the pharmaceutically acceptable acid is benzoic acid. In some embodiments, the pharmaceutically acceptable acid is salicylic acid. In some embodiments, the pharmaceutically acceptable acid is gallic acid.

[0101] The present disclosure encompasses the preparation and use of solvates of USP1 inhibitors and / or PARP1-selective inhibitors. Solvates typically do not significantly alter the physiological activity or toxicity of the compounds and thus may function as pharmacological equivalents. As used herein, the term "solvate" refers to a combination, physical association, and / or solvation of a disclosed USP1 inhibitor or PARP1-selective inhibitor with solvent molecules, such as a disolvate, monosolvate, or hemisolvate, in which the ratio of solvent molecules to the disclosed compound is about 2:1, about 1:1, or about 1:2, respectively. This physical association includes varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain cases, the solvate may be isolated, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Thus, "solvate" encompasses both solution-phase and isolatable solvates. The disclosed USP1 inhibitors or PARP1 selective inhibitors may exist as solvates with pharmaceutically acceptable solvents such as water, methanol, ethanol, etc., and the present disclosure is intended to encompass both solvated and unsolvated forms of the disclosed USP1 inhibitors and / or PARP1 selective inhibitors. One type of solvate is a hydrate. "Hydrate" refers to a specific subgroup of solvates in which the solvent molecule is water. "Anhydrous" as applied to a compound refers to a solid state in which the compound does not contain structural water in the crystal lattice. Solvates can typically function as pharmacological equivalents. Preparation of solvates is known in the art. See, for example, M. Caira et al., J. Pharmaceut. Sci., 93(3):601-611 (2004), which describes the preparation of ethyl acetate and water solvates of fluconazole. The preparation of similar solvates, hemisolvates, hydrates, etc. is described in EC van Tonder et al., AAPS Pharm. Sci. Tech., 5(1):Article 12 (2004), and AL Bingham et al., Chem. Commun. 603-604 (2001).A typical, non-limiting process for preparing a solvate includes dissolving a disclosed USP1 inhibitor or PARP1 selective inhibitor in a desired solvent (organic solvent, water, or a mixture thereof) at a temperature above 20°C to about 25°C, then cooling the solution at a rate sufficient to form crystals, and isolating the crystals by known methods, for example, filtration. Analytical techniques such as infrared spectroscopy can be used to confirm the presence of solvent in the solvate crystals.

[0102] In some embodiments of the present disclosure, the USP1 inhibitor and / or PARP1 selective inhibitor is deuterated. In some embodiments, the USP1 inhibitor and / or PARP1 selective inhibitor is partially or fully deuterated, i.e., one or more hydrogen atoms are replaced with deuterium atoms.

[0103] As used herein, "treatment" refers to an approach to obtain beneficial or desired clinical results. As used herein, "treatment" encompasses the administration or application of a therapeutic agent to a disease in a mammal, including a human. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, any one or more of the following: alleviation of one or more symptoms, reduction in the extent of the disease, prevention or delay of disease spread (e.g., metastasis), prevention or delay of disease recurrence, delay or slowing of disease progression, amelioration of the disease state, inhibition of the disease or disease progression, inhibition or slowing of the disease or its progression, prevention of its onset, and remission (whether partial or total). "Treatment" also encompasses alleviation of the pathological consequences of a proliferative disease. The methods provided herein contemplate any one or more of these aspects of treatment. In line with the above, the term treatment does not require 100% elimination of all aspects of the disorder.

[0104] In the context of cancer, the term "treating" includes, but is not limited to, inhibiting the growth of cancer cells, inhibiting the replication of cancer cells, lessening the overall tumor burden, and delaying, halting, or slowing the growth, progression, or metastasis of tumors.

[0105] As used herein, "delaying" means to defer, hinder, slow, retard, stabilize, suppress, and / or postpone the onset or progression of a disease (such as cancer). This delay can be for varying lengths of time, depending on the medical history of the disease and / or individual being treated.

[0106] The "therapeutically effective amount" of a substance can vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance to induce a desired response in the individual. A therapeutically effective amount is also an amount in which the therapeutically beneficial effects outweigh the toxic or harmful effects of the substance. A therapeutically effective amount can be delivered in one or more administrations. A therapeutically effective amount refers to an amount effective at the dosage and for the period of time necessary to achieve the desired therapeutic effect.

[0107] As used herein, the term "combination," "therapeutic combination," "combination composition," "combination therapy," or "pharmaceutical combination" may include a fixed combination in a single dosage unit, a separate dosage unit, or a kit of parts or instructions for combined administration, in which the USP1 inhibitor and the PARP1 selective inhibitor can be administered simultaneously or separately within a time interval. The combined pharmaceutical composition can be adapted for simultaneous administration, separate administration, or sequential administration.

[0108] Combination therapy can provide a "synergistic effect" and prove to be "synergistic." That is, the effect achieved when the active ingredients are used together is greater than the sum of the effects resulting from using the compounds separately. A synergistic effect can include a significant reduction in the effective amount of the combination of two active ingredients compared to the effective amount of each active ingredient when administered separately. A synergistic effect can also include a reduction in the toxicity of the combination of two active ingredients compared to the toxicity of each active ingredient when administered separately. A synergistic effect can also be an effect that cannot be achieved by administering either active ingredient as a single agent. Synergistic effects can include, but are not limited to, the treatment of cancer by reducing tumor size, inhibiting tumor growth, or prolonging the survival of a subject. Synergistic effects can also include reducing cancer cell viability, inducing cancer cell death, and inhibiting or delaying cancer cell growth. A synergistic effect can be achieved, for example, when the active ingredients are (1) co-formulated and administered or delivered simultaneously in a combined unit dosage formulation; (2) delivered sequentially, alternately, or concurrently as separate formulations; or (3) by other regimens. When delivered in alternation therapy, a synergistic effect is achieved when the compounds are administered or delivered sequentially.

[0109] The synergistic interaction between a USP1 inhibitor and a PARP1 selective inhibitor can be determined based on the results obtained from the assays described herein. For example, the combined effect can be evaluated using the Bliss independence model. The Bliss score quantifies the degree of potentiation by a single agent, and a Bliss score of >0 indicates greater than simple additivity. In some embodiments, a Bliss score of greater than 10 indicates a strong synergistic effect. In some embodiments, a score of 6 or greater indicates a synergistic effect. In some embodiments, the Bliss score is about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, or about 25.

[0110] As used herein, "homologous recombination deficiency score" or "HRD score" refers to an algorithmic assessment of three measures of tumor genomic instability: loss of heterozygosity, telomeric allele imbalance, and large-scale state transitions.

[0111] The terms "administer," "administering," "administration," and the like refer to methods that can be used to enable delivery of a therapeutic agent to a desired site of biological action. Administration techniques that can be employed with the agents and methods described herein are described, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. Administration of two or more therapeutic agents includes simultaneous (concurrent) administration in any order and sequential administration.

[0112] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a formulation that is in a form that allows the biological activity of the active ingredient to be effective and that does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. Such formulations may be sterile.

[0113] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0114] "Pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid, or carrier conventionally used in the art for use with therapeutic agents that together comprise a "pharmaceutical composition" for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations employed and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is appropriate for the formulation being used.

[0115] A "sterile" formulation is aseptic or essentially free of living microorganisms and their spores.

[0116] The term "container" means a container and closure suitable for storing, transporting, dispensing, and / or handling a pharmaceutical product.

[0117] "Insert" or "package insert" means the information that accompanies a drug product and provides instructions on how to administer the drug, along with safety and effectiveness data necessary for physicians, pharmacists, and patients to make informed decisions regarding the drug's use. Generally, package inserts are considered the drug's "label."

[0118] As used herein, the term "disease" or "condition" or "disorder" refers to a condition for which treatment is necessary and / or desirable, and refers to a disorder and / or abnormality that is generally considered to be a pathological state or function and may manifest itself in the form of specific signs, symptoms, and / or dysfunction. As described below, the combination of a USP1 inhibitor and a PARP1-selective inhibitor of the present disclosure can be used to treat diseases and conditions, such as proliferative diseases, in which inhibition of USP1 and / or PARP1 would be beneficial.

[0119] The terms "polypeptide" and "protein" are used interchangeably to refer to polymers of amino acid residues and are not limited to a minimum length. Such polymers of amino acid residues may contain natural or non-natural amino acid residues, and include, but are not limited to, peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. Both full-length proteins and fragments thereof are encompassed by this definition. These terms also include post-expression modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, and the like. Furthermore, for purposes of this disclosure, "polypeptide" refers to proteins containing modifications such as deletions, additions, and substitutions (generally conservative in nature) to the native sequence, so long as the protein maintains the desired activity. These modifications may be deliberate, such as through site-directed mutagenesis, or may be accidental, such as through mutations of hosts producing the protein or errors due to PCR amplification.

[0120] As used herein, "USP1" and "ubiquitin-specific processing protease 1" refer to a native polypeptide or a USP1-encoding polynucleotide. The term "USP1" encompasses the "full-length" unprocessed USP1 polypeptide and forms of USP1 that result from intracellular processing (e.g., removal of the signal peptide). The term also encompasses naturally occurring variants of USP1, such as those encoded by splice variants and allelic variants. The USP1 polypeptides described herein can be isolated from various sources, such as human tissue types or another source, or prepared by recombinant or synthetic methods. Human USP1 sequences are known and include, for example, the sequence publicly available as UniProt number O94782 (including isoforms). As used herein, the term "human USP1 protein" refers to SEQ ID NO: 1: [ka] It refers to a USP1 protein comprising the amino acid sequence set forth in

[0121] USP1 is a deubiquitinating enzyme that acts as part of a complex with UAF1. The "deubiquitinating enzyme activity" of USP1 includes its ability to deubiquitinate as part of the USP1-UAF1 complex.

[0122] As used herein, "PARP" or "PARP protein" refers to one or more enzymes in the poly(ADP-ribose) polymerase family of enzymes. This family includes enzymes that have the ability to catalyze the transfer of ADP-ribose to target proteins (poly-ADP-ribosylation). The PARP family includes at least 18 members, including PARP1, PARP2, and PARP3, that are encoded by different genes and share homology in a conserved catalytic domain.

[0123] As used herein, the term "PARP1 selective inhibitor" refers to an inhibitor that is selective for PARP1 over PARP2. In some embodiments, the PARP1 selective inhibitor is 500-fold more selective for PARP1 than PARP2. An example of a PARP1 selective inhibitor is AZD5305.

[0124] The term "specifically binds" is well understood in the art, and methods for determining such specific binding are also well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular protein or domain of a protein more frequently, more rapidly, for a longer period, and / or with higher affinity than another protein or domain. It should be understood that a molecule that specifically or preferentially binds to a first protein or domain may or may not specifically or preferentially bind to a second protein or domain. Thus, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Generally, reference to binding implies preferential binding, but this is not necessarily the case. For example, a USP1 inhibitor that specifically binds to USP1, UAF1 and / or the USP1-UAF1 complex may not bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or may bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with reduced affinity compared to binding to USP1.

[0125] The terms "reduction" or "reduce" or "inhibition" or "inhibiting" refer to a decrease or cessation of a phenotypic trait, or a decrease or cessation of the incidence, degree, or likelihood of that trait. "Reduce" or "inhibiting" refers to a decrease, reduction, or cessation of an activity, function, and / or amount compared to a reference. In some embodiments, "reduce" or "inhibiting" refers to the ability to cause an overall reduction of 20% or more. In some embodiments, "reduce" or "inhibiting" refers to the ability to cause an overall reduction of 50% or more. In some embodiments, "reduce" or "inhibiting" refers to the ability to cause an overall reduction of 75%, 85%, 90%, 95%, or more. In some embodiments, the amount is inhibited or reduced over a period of time compared to a control over the same period of time.

[0126] In some embodiments, inhibiting the USP1 protein refers to inhibiting one or more activities or functions of the USP1 protein. It should be understood that one or more activities or functions of the USP1 protein can be inhibited in vitro or in vivo. Non-limiting examples of USP1 activities and functions include deubiquitinase activity and complex formation with UAF1, and are described herein. Exemplary inhibition levels of one or more USP1 protein activities include at least 10% inhibition, at least 20% inhibition, at least 30% inhibition, at least 40% inhibition, at least 50% inhibition, at least 60% inhibition, at least 70% inhibition, at least 80% inhibition, at least 90% inhibition, and up to 100% inhibition.

[0127] In some embodiments, inhibiting PARP protein refers to inhibiting one or more activities or functions of PARP protein.It should be understood that one or more activities or functions of PARP protein can be inhibited in vitro or in vivo.Non-limiting examples of PARP activity and function are described herein.The exemplary inhibition level of one or more activities of PARP protein includes at least 10% inhibition, at least 20% inhibition, at least 30% inhibition, at least 40% inhibition, at least 50% inhibition, at least 60% inhibition, at least 70% inhibition, at least 80% inhibition, at least 90% inhibition and up to 100% inhibition.

[0128] The terms "individual" or "subject" are used interchangeably herein to refer to animals, e.g., mammals such as humans. In some cases, methods are provided for treating mammals, including but not limited to humans, rodents, apes, cats, dogs, cows, pigs, sheep, goats, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets. In some instances, "individual" or "subject" refers to an individual or subject in need of treatment for a disease or disorder. In some cases, the subject receiving treatment may be a patient who presents with the fact that the subject has been identified as having a disorder related to the treatment or has been identified as being at particular risk for the disorder.

[0129] As used herein, the terms "cancer" and "tumor" refer to or describe a physiological condition in a mammal in which a cell population is characterized by uncontrolled cell proliferation. These terms include solid cancers and hematological / lymphatic cancers. Examples of cancers include, but are not limited to, DNA damage repair pathway-deficient cancers. Further examples of cancers include, but are not limited to, ovarian cancer, breast cancer (including triple-negative breast cancer), non-small cell lung cancer (NSCLC), and osteosarcoma. The cancer may be BRCA1 wild-type or BRCA2 wild-type. The cancer may also be BRCA1 or BRCA2 mutant. The cancer may further be PARP inhibitor-resistant or refractory cancer, or PARP inhibitor-resistant or refractory BRCA1 or BRCA2 mutant cancer.

[0130] As used herein, the term "loss-of-function" mutation refers to a mutation that results in the absence of a gene, reduced gene expression, or the production of a gene product (e.g., a protein) with reduced or no activity. Loss-of-function mutations include, for example, missense mutations, nucleotide insertions, nucleotide deletions, and gene deletions. Loss-of-function mutations also include negative mutations. Therefore, cancer cells that have a loss-of-function mutation in the gene encoding p53 include cancer cells that have a missense mutation in the gene encoding p53 and cancer cells that lack the gene encoding p53.

[0131] USP1 inhibitors USP1 inhibitors are disclosed, for example, in WO 2020 / 132269 and WO 2022 / 094096 (each incorporated herein by reference in its entirety).

[0132] In some embodiments, the ubiquitin-specific processing protease 1 (USP1) inhibitors of the present disclosure: Formula I: [ka] Formula II: [ka] Formula III: [ka] or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In some embodiments, the ubiquitin-specific processing protease 1 (USP1) inhibitor comprises a compound of: (a) Formula I: [ka] (b) Formula II: [ka] (c) Formula III: [ka] and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof.

[0133] The chemical name of the USP1 inhibitor of Formula I is 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, as described in U.S. Patent Application Publication No. 2021 / 0115049. This compound is also referred to herein as Formula I.

[0134] The chemical name of the USP1 inhibitor I of Formula I is 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, as described in U.S. Patent Application Publication No. 2021 / 0115049. This compound is also referred to herein as Formula II.

[0135] The chemical name of USP1 inhibitor II of Formula I is 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, as described in U.S. Patent Application Publication No. 2021 / 0115049, which is incorporated herein by reference in its entirety. This compound is also referred to herein as Formula III.

[0136] In some embodiments, the USP1 inhibitor for the uses and methods provided herein is a solid form that is a pharmaceutically acceptable salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I). ​​In some embodiments, the pharmaceutically acceptable salt is formed between 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I) and a pharmaceutically acceptable acid. In some embodiments, the pharmaceutically acceptable acid is gentisic acid.

[0137] In some embodiments, the gentisate salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I gentisate salt) is in an amorphous form. In some embodiments, the amorphous form is substantially free of other polymorphic forms. In some embodiments, the amorphous form has a polymorphic purity of at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 86%, or at least 87%, or at least 88%, or at least 89%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%.

[0138] In another embodiment, the USP1 inhibitor for the uses and methods provided herein is a solid form that is a co-crystal of a pharmaceutically acceptable acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I). In some embodiments, the co-crystal is a mixture of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I) and 1-hydroxy-2-naphthoic acid, 4-aminosalicylic acid, ascorbic acid, adipic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, trans-cinnamic acid, citric acid, ethanedisulfonic acid, fumaric acid, galactaric acid, gentisic acid, gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycolic acid, hexanoic acid, hippuric acid, The pharmaceutically acceptable acid is formed between a pharmaceutically acceptable acid selected from the group consisting of hydrobromic acid, hydrochloric acid, lactic acid, maleic acid, L-malic acid, malonic acid, R-mandelic acid, methanesulfonic acid, mucic acid, naphthalenesulfonic acid, nicotinic acid, oxalic acid, palmitic acid, p-toluenesulfonic acid, phosphoric acid, propionic acid, saccharin, salicylic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, vanillic acid, vanillin, ethyl maltol, gallic acid, gallic acid ethyl ester, 4-hydroxybenzoic acid, 4-hydroxybenzoic acid methyl ester, 3,4,5-trihydroxybenzoic acid, nicotinamide, L-proline, and D-sorbitol. In some embodiments, the pharmaceutically acceptable acid is selected from the group consisting of gentisic acid, benzoic acid, salicylic acid, and gallic acid. In some embodiments, the pharmaceutically acceptable acid is gentisic acid. Co-crystals of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine are disclosed, for example, in WO 2022 / 094096, the entire contents of which are incorporated herein by reference.

[0139] In some embodiments, the co-crystal of gentisic acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I gentisic acid co-crystal) is in an amorphous form. In some embodiments, the amorphous form is substantially free of other polymorphic forms. In some embodiments, the amorphous form has a polymorphic purity of at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 86%, or at least 87%, or at least 88%, or at least 89%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%.

[0140] In some embodiments, the gentisate salt of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I gentisate salt) is in a crystalline form.

[0141] In some embodiments, the pharmaceutically acceptable co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I) is a gentisic acid co-crystal.

[0142] In some embodiments, the pharmaceutically acceptable co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine of Formula I is a benzoic acid co-crystal.

[0143] In some embodiments, the pharmaceutically acceptable co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine of Formula I is a salicylic acid co-crystal.

[0144] In some embodiments, the gentisic acid co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula I gentisic acid co-crystal) is in crystalline Form 2 (Formula I gentisic acid co-crystal, Form 2 or "Form 2"). Formula I gentisic acid co-crystal, Form 2 is characterized by an XRPD pattern with peaks at 16.6±0.2 degrees 2θ, 18.7±0.2 degrees 2θ, and 22.5±0.2 degrees 2θ.

[0145] In some embodiments, crystalline Form 2 is anhydrous.

[0146] In some embodiments, the melting point of crystalline form 2 is about 184°C to about 190°C. In some embodiments, the melting point of crystalline form 2 is about 186°C to about 188°C. In some embodiments, the melting point of crystalline form 2 is about 187°C.

[0147] In some embodiments, crystalline form 2 is characterized by an XRPD pattern with peaks at 16.6±0.2°2θ, 18.7±0.2°2θ, and 22.5±0.2°2θ as measured with CuKα radiation. In another embodiment, crystalline form 2 is characterized by an XRPD pattern with peaks at 16.6±0.2°2θ, 18.7±0.2°2θ, 22.3±0.2°2θ, and 22.5±0.2°2θ as measured with CuKα radiation. In some embodiments, crystalline form 2 is characterized by an XRPD pattern with peaks at 16.6±0.2°2θ, 18.7±0.2°2θ, 22.3±0.2°2θ, 22.5±0.2°2θ, and 26.0±0.2°2θ as measured with CuKα radiation. In some embodiments, crystalline Form 2 is characterized by an XRPD pattern with peaks at 16.6±0.2 degrees 2θ, 18.7±0.2 degrees 2θ, 20.8±0.2 degrees 2θ, 22.3±0.2 degrees 2θ, 22.5±0.2 degrees 2θ, and 26.0±0.2 degrees 2θ, as measured with CuKα radiation.

[0148] In some embodiments, crystalline Form 2 is characterized by an XRPD pattern substantially as shown in Figure 2. In some embodiments, crystalline Form 2 is characterized by 3 or more, 4 or more, 5 or more, or 6 or more XRPD peaks listed in Table 1(a and b) below: [Table 1a] [Table 1b]

[0149] In some embodiments, crystalline Form 2 is characterized by an endothermic peak at about 181° C. to about 191° C., or about 183° C. to about 189° C., or about 185° C. to about 187° C., as determined by DSC. In some embodiments, crystalline Form 2 is characterized by an endothermic peak at about 186.0° C., as determined by DSC.

[0150] In some embodiments, crystalline Form 2 is characterized by a DSC profile substantially as shown in FIG.

[0151] In some embodiments, crystalline Form 2 is characterized by a loss of about 2.5 wt% to about 3.5 wt% between room temperature and about 170° C. In some embodiments, crystalline Form 2 is characterized by a loss of about 3.0 wt% to about 3.4 wt% between room temperature and about 170° C. In some embodiments, crystalline Form 2 is characterized by a loss of about 3.17 wt% between room temperature and about 170° C.

[0152] In some embodiments, crystalline Form 2 is characterized by a TGA profile substantially as shown in FIG.

[0153] In some embodiments, crystalline Form 2 is characterized by at least two of the following: a) the XRPD pattern shown in FIG. 2; b) the DSC profile shown in FIG. 3; or c) the TGA profile shown in FIG. 3.

[0154] In some embodiments, crystalline Form 2 has a unit cell indexed as monoclinic.

[0155] In some embodiments, crystalline Form 2 has a unit cell with an a value of about 11.113 Å, a b value of about 12.356 Å, and a c value of about 24.048 Å. 3 It has a unit cell with a volume of

[0156] Unit cell parameters of crystalline form 2 shown in Table 2. [Table 2]

[0157] In some embodiments, crystalline Form 2 is substantially free of other polymorphic forms. In some embodiments, crystalline Form 2 has a polymorphic purity of at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 86%, or at least 87%, or at least 88%, or at least 89%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%.

[0158] In some embodiments, the USP1 inhibitor comprises a mixture comprising crystalline Form 2 and a second solid form of Formula I. In some embodiments, the second solid form of Formula I is crystalline Form A. In some embodiments, the USP1 inhibitor comprises a mixture comprising a majority of crystalline Form 2 compared to other solid forms of Formula I. Suitable solid forms of Formula I are described in WO 2022 / 094096, which is incorporated herein by reference in its entirety.

[0159] In another embodiment, the USP1 inhibitor is of Formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In another embodiment, the USP1 inhibitor is a solid form of Formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

[0160] In some embodiments, the USP1 inhibitor for the uses and methods provided herein is a solid form that is a co-crystal of a pharmaceutically acceptable acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula II). In some embodiments, the pharmaceutically acceptable salt is formed between 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula II) and a pharmaceutically acceptable acid.

[0161] Suitable solid forms of Formula II are described in WO 2022 / 094096, which is incorporated herein by reference in its entirety.

[0162] In another embodiment, the USP1 inhibitor is of Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In another embodiment, the USP1 inhibitor is of Formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

[0163] In some embodiments, the USP1 inhibitor for the uses and methods provided herein is a solid form that is a co-crystal of a pharmaceutically acceptable acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula III). In some embodiments, the pharmaceutically acceptable salt is formed between 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Formula III) and a pharmaceutically acceptable acid.

[0164] Suitable solid forms of Formula III are described in WO 2022 / 094096, which is incorporated herein by reference in its entirety.

[0165] The present disclosure encompasses the preparation and use of salts of USP1 inhibitors, including non-toxic pharmaceutically acceptable salts. Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts and base salts. Pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium salts, potassium salts, cesium salts, and the like; alkaline earth metal salts such as calcium salts, magnesium salts, and the like; organic amine salts such as triethylamine salts, pyridine salts, picoline salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, and the like; inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulfate, and the like; organic acid salts such as citrate, lactate, tartrate, maleate, fumarate, mandelate, acetate, dichloroacetate, trifluoroacetate, oxalate, formate, and the like; sulfonate salts such as methanesulfonate, benzenesulfonate, p-toluenesulfonate, and the like; and amino acid salts such as alginate, aspartate, glutamate, and the like. As used herein, the term "pharmaceutically acceptable salt" refers to any salt, obtained, for example, by reaction of a USP1 inhibitor of the present disclosure with an acid or base, that is physiologically tolerated in a target patient (e.g., a mammal, e.g., a human).

[0166] Methods for synthesizing USP inhibitors, including those of Formula I, II, and III, are described in U.S. Patent Application Publication No. 2021 / 0115049, which is incorporated herein by reference in its entirety. Methods for preparing salts and polymorphic forms of USP inhibitors, including those of Formula I, II, and III, are described in PCT Publication No. 2022 / 094096, which is incorporated herein by reference in its entirety.

[0167] In various embodiments, the USP1 inhibitor reduces the level of USP1 protein and / or inhibits or reduces at least one biological activity of USP1 protein.

[0168] In some embodiments, the USP1 inhibitor specifically binds to USP1 protein. In some embodiments, the USP1 inhibitor specifically binds to USP1 protein in the USP1-UAF1 complex. In some embodiments, the USP1 inhibitor specifically binds to USP1 mRNA. In some embodiments, the USP1 inhibitor specifically binds to USP1 protein (alone or in the USP1-UAF1 complex) or USP1 mRNA. In some embodiments, the USP1 inhibitor specifically binds to UAF1 (alone or in the USP1-UAF1 complex) and inhibits or reduces the formation or activity of the USP1-UAF1 complex.

[0169] In some embodiments, the USP1 inhibitor reduces the formation of the USP1-UAF1 complex. In some embodiments, the USP1 inhibitor reduces the activity of the USP1-UAF1 complex. In some embodiments, the USP1 inhibitor reduces the deubiquitinase activity of USP1. In some embodiments, the USP1 inhibitor increases monoubiquitinated PCNA. In some embodiments, the USP1 inhibitor increases monoubiquitinated FANCD2. In some embodiments, the USP1 inhibitor increases monoubiquitinated FANCI.

[0170] In some embodiments, the USP1 inhibitor does not bind to other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or binds to deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with an affinity that is at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold lower than the affinity for USP1 (i.e., the K of the USP1 inhibitor for other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1)). D is the K D at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold greater than the

[0171] In some embodiments, the USP1 inhibitor inhibits USP1 deubiquitinase activity with an IC50 of less than about 50 nM, about 50 nM to about 200 nM, about 200 nM to about 2 pM, or greater than 2 pM, as measured, for example, using the assay disclosed in U.S. Patent Application Publication No. 2017 / 0145012, or with an IC50 of 50 nM to 1000 nM, as measured, for example, using the assay described in Liang et al., Nat Chem Biol 10: 289-304 (2014). In some embodiments, the USP1 inhibitor inhibits USP1 deubiquitinase activity with an IC50 as measured using the assay disclosed in Chen, et al., Chem Biol., 18(11):1390-1400 (2011). In some embodiments, the USP1 inhibitor does not inhibit the activity of other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or inhibits the activity of other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with an IC50 that is at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold greater than the IC50 for inhibiting USP1 deubiquitinase activity.

[0172] In some embodiments, the USP1 inhibitors of the present disclosure bind to USP1 protein with an affinity ranging from 1 pM to 100 μM, or from 1 pM to 1 μM, or from 1 pM to 500 nM, or from 1 pM to 100 nM. In some embodiments, the USP1 inhibitors of the present disclosure bind to USP1 protein with an affinity of about 1 pM to about 100 μM, about 1 nM to about 100 μM, about 1 μM to about 100 μM, about 1 μM to about 50 μM, about 1 μM to about 40 μM, about 1 μM to about 30 μM, about 1 μM to about 20 μM, or about 1 μM to about 10 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the USP1 inhibitors of the present disclosure are from about 100 nM to about 1 μM, from about 100 nM to about 900 nM, from about 100 nM to about 800 nM, from about 100 nM to about 700 nM, from about 100 nM to about 600 nM, from about 100 nM to about 500 nM, from about 100 nM to about 400 nM, from about 100 nM to about 300 nM, from about 100 nM to about 200 nM, from about 200 nM to about 1 μM, or from about 300 nM to about 400 nM. and binds to the USP1 protein with an affinity of about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, about 900 nM, about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM.In some embodiments, the USP1 inhibitors of the present disclosure are from about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, about 50 nM to about The antibody binds to the USP1 protein with an affinity of 100 nM, about 60 nM to about 100 nM, about 70 nM to about 100 nM, about 80 nM to about 100 nM, about 90 nM to about 100 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, or about 100 nM.

[0173] In some embodiments, the USP1 inhibitors of the present disclosure bind to USP1 protein with an affinity of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the USP1 inhibitors bind to USP1 protein with an affinity of less than 1 nM.

[0174] In some embodiments, the USP1 inhibitors of the present disclosure inhibit USP1 activity with an IC50 of 1 pM to 100 μM, or 1 pM to 1 μM, or 1 pM to 500 nM, or 1 pM to 100 nM. In some embodiments, the USP1 inhibitor inhibits USP1 activity with an IC50 of about 1 pM to about 100 μM, about 1 nM to about 100 μM, about 1 μM to about 100 μM, about 1 μM to about 50 μM, about 1 μM to about 40 μM, about 1 μM to about 30 μM, about 1 μM to about 20 μM, or about 1 μM to about 10 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the USP1 inhibitor is from about 100 nM to about 1 μM, from about 100 nM to about 900 nM, from about 100 nM to about 800 nM, from about 100 nM to about 700 nM, from about 100 nM to about 600 nM, from about 100 nM to about 500 nM, from about 100 nM to about 400 nM, from about 100 nM to about 300 nM, from about 100 nM to about 200 nM, from about 200 nM to about 1 μM, or from about 300 nM to about 500 nM. Inhibits USP1 activity with an IC50 of about 1 μM, about 400 nM to about 1 μM, about 500 nM to about 1 μM, about 600 nM to about 1 μM, about 700 nM to about 1 μM, about 800 nM to about 1 μM, about 900 nM to about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM.

[0175] In some embodiments, the USP1 inhibitor of the present disclosure is at a concentration of about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, about 50 nM to about 60 nM, 1 nM to about 70 nM, 1 nM to about 80 nM, 1 nM to about 90 nM, 1 nM to about 100 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, or about 50 nM to about 60 nM. Inhibits USP1 activity with an IC50 of about 100 nM, about 60 nM to about 100 nM, about 70 nM to about 100 nM, about 80 nM to about 100 nM, about 90 nM to about 100 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, or about 100 nM. In some embodiments, the USP1 inhibitor inhibits USP1 activity with an IC50 of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the USP1 inhibitor inhibits USP1 activity with an IC50 of less than 1 nM.

[0176] Other exemplary USP1 inhibitors are disclosed, for example, in WO 2020 / 132269 and U.S. Provisional Patent Application No. 62 / 857,986 (each of which is incorporated herein by reference in its entirety).

[0177] Exemplary Assay for Inhibition of USP1 Any suitable assay in the art can be used to determine activity, detect results or effects, or determine efficacy. See, e.g., U.S. Patent Application Publication No. 2021 / 0115049, incorporated herein by reference in its entirety.

[0178] In some cases, the method for determining whether a USP1 inhibitor compound inhibits USP1 deubiquitinase activity involves measuring the change in mass upon cleavage of the deubiquitinase bond to diubiquitin. For example, ubiquitin aldehyde and ubiquitin vinyl sulfone form an irreversible covalent bond with the deubiquitinase, resulting in an observable mass change for the deubiquitinase. Similarly, cleavage of diubiquitin results in an observable mass change.

[0179] In some cases, methods for determining whether a USP1 inhibitor compound inhibits USP1 deubiquitinase activity include an increase in luminescence or fluorescence upon cleavage, which can be monitored, for example, on a plate reader. Such assays can use ubiquitin linked to a fluorophore via a linker linkage, such as ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) or ubiquitin-rhodamine 110. Such assays can also use diubiquitins containing an isopeptide bond. An exemplary diubiquitin may contain a fluorophore on one ubiquitin and a quencher on the other, such that fluorescence increases as the diubiquitin is cleaved. Such assays can also use enzyme-linked systems in which ubiquitin is coupled to an enzyme that is only active in producing a fluorescent enzyme product upon release from the ubiquitin.

[0180] PARP1 selective inhibitor In various embodiments, the PARP1 selective inhibitors of the present disclosure reduce the level of PARP1 protein and / or inhibit or reduce at least one biological activity of PARP1 protein.

[0181] AZD5305 (5-[4-[(7-ethyl-6-oxo-5H-1,5-naphthyridin-3-yl)methyl]piperazin-1-yl]-N-methylpyridine-2-carboxamide) is a small molecule drug that acts by selectively inhibiting and trapping PARP1 at sites of DNA single-strand breaks (SSBs). This prevents DNA repair and leads to the generation of more damaging DNA double-strand breaks (DSBs) during DNA replication when the DNA replication machinery collides with the PARP1-DNA noncovalent complex. In situations where accurate DNA repair pathways are active, such as in cells with sufficient homologous recombination repair (HRR), DSBs are accurately repaired. In contrast, in cells with defective repair pathways, such as those carrying deleterious mutations in the BRCA genes, AZD5305 treatment leads to the selective accumulation of genomic instability, ultimately resulting in the selective death of cancer cells while sparing normal cells. See PCT Application Publication No. 2022 / 074124, incorporated herein by reference.

[0182] The term "AZ05305" refers to a compound having the chemical name 5-[4-[(7-ethyl-6-oxo-5H-1,5-naphthyridin-3-yl)methyl]piperazin-1-yl]-N-methyl-pyridine-2-carboxamide and the structure shown below: [ka] PCT Publication No. 2022 / 074124 discloses the preparation of AZD5305. In some embodiments, the free base of AZD5305 is administered to a subject. In some embodiments, a pharmaceutically acceptable salt of AZD5305 is administered to a subject. In some embodiments, crystalline AZD5305 is administered to a subject. In some embodiments, crystalline form A of AZD5305 is administered to a subject.

[0183] In one embodiment, a PARP1-selective inhibitor is used in an anti-cancer combination therapy with a USP1 inhibitor of the present disclosure. In addition to the PARP1-selective inhibitor and the USP1 inhibitor, other therapies can be used before, during, or after the combination therapy.

[0184] Exemplary Assays for Inhibition of PARP The present disclosure provides compounds that are active in inhibiting the activity of PARP1. To determine activity, detect results or effects, or determine effectiveness, suitable assays in the art can be used. For example, see Dillon, et al., JBS., 8(3), 347-352 (2003); U.S. Patent No. 9,566,276. In some embodiments, PARP1 selective inhibitors selectively inhibit and capture PARP1 in cells, as determined by using a PARP capture assay, for example, as disclosed in Illuzzi G., et al., Clinical Cancer Research, doi: 10.1158 / 1078-0432.CCR-22-0301 (2022). In some embodiments, PARP1 selective inhibitors do not efficiently capture PARP1 in cells.

[0185] In some embodiments, the PARP1 selective inhibitors of the present disclosure inhibit PARP1 activity with an IC50 of less than about 50 nM, between about 50 nM and about 200 nM, between about 200 nM and about 2 pM, or greater than 2 pM.

[0186] In some embodiments, the PARP1 selective inhibitors of the present disclosure bind to PARP1 protein with an affinity ranging from 1 pM to 100 μM, or from 1 pM to 1 μM, or from 1 pM to 500 nM, or from 1 pM to 100 nM. In some embodiments, the PARP1 selective inhibitor of the present disclosure binds to PARP1 protein with an affinity of about 1 pM to about 100 μM, about 1 nM to about 100 μM, about 1 μM to about 100 μM, about 1 μM to about 50 μM, about 1 μM to about 40 μM, about 1 μM to about 30 μM, about 1 μM to about 20 μM, or about 1 μM to about 10 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the PARP1 selective inhibitor of the present disclosure has a concentration of about 100 nM to about 1 μM, about 100 nM to about 900 nM, about 100 nM to about 800 nM, about 100 nM to about 700 nM, about 100 nM to about 600 nM, about 100 nM to about 500 nM, about 100 nM to about 400 nM, about 100 nM to about 300 nM, about 100 nM to about 200 nM, about 200 nM to about 1 μM, about 30 It binds to PARP1 protein with an affinity of 0 nM to about 1 μM, about 400 nM to about 1 μM, about 500 nM to about 1 μM, about 600 nM to about 1 μM, about 700 nM to about 1 μM, about 800 nM to about 1 μM, about 900 nM to about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM.In some embodiments, the PARP1 selective inhibitor of the present disclosure has a concentration of about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, about 50 nM to The PARP1 selective inhibitors of the present disclosure bind to PARP1 protein with an affinity of about 100 nM, about 60 nM to about 100 nM, about 70 nM to about 100 nM, about 80 nM to about 100 nM, about 90 nM to about 100 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, or about 100 nM. In some embodiments, the PARP1 selective inhibitors of the present disclosure bind to PARP1 protein with an affinity of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the PARP1 selective inhibitors of the present disclosure bind to PARP1 protein with an affinity of less than 1 nM.

[0187] In some embodiments, the PARP1 selective inhibitors of the present disclosure inhibit PARP1 activity with an IC50 of 1 pM to 100 μM, or 1 pM to 1 μM, or 1 pM to 500 nM, or 1 pM to 100 nM. In some embodiments, the PARP1 selective inhibitors of the present disclosure inhibit PARP1 activity with an IC50 of about 1 pM to about 100 μM, about 1 nM to about 100 μM, about 1 μM to about 100 μM, about 1 μM to about 50 μM, about 1 μM to about 40 μM, about 1 μM to about 30 μM, about 1 μM to about 20 μM, or about 1 μM to about 10 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the PARP1 selective inhibitor of the present disclosure has a concentration of about 100 nM to about 1 μM, about 100 nM to about 900 nM, about 100 nM to about 800 nM, about 100 nM to about 700 nM, about 100 nM to about 600 nM, about 100 nM to about 500 nM, about 100 nM to about 400 nM, about 100 nM to about 300 nM, about 100 nM to about 200 nM, about 200 nM to about 1 μM, about 3 Inhibits PARP1 activity with an IC50 of about 100 nM to about 1 μM, about 400 nM to about 1 μM, about 500 nM to about 1 μM, about 600 nM to about 1 μM, about 700 nM to about 1 μM, about 800 nM to about 1 μM, about 900 nM to about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM.In some embodiments, the PARP1 selective inhibitor of the present disclosure has a concentration of about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, about 50 nM Inhibits PARP1 activity with an IC50 of about 100 nM, about 60 nM to about 100 nM, about 70 nM to about 100 nM, about 80 nM to about 100 nM, about 90 nM to about 100 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, or about 100 nM. In some embodiments, the PARP1 selective inhibitors of the present disclosure inhibit PARP1 activity with an IC50 of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the PARP1 selective inhibitors of the present disclosure inhibit PARP1 activity with an IC50 of less than 1 nM.

[0188] In some embodiments, the PARP1 selective inhibitor is at least 500-fold more selective for PARP1 than for PARP2.

[0189] How to use Because the combinations of the present disclosure are inhibitors of USP1 protein and PARP1 protein, the present disclosure provides a method of inhibiting USP1 protein and / or PARP1 protein, the method comprising contacting USP1 and / or PARP1 protein, or a composition comprising USP1 and / or PARP1 protein, with one or more combinations of the present disclosure.

[0190] Because the combinations of the present disclosure are inhibitors of USP1 and PARP1 proteins, these compounds can be used to treat a number of diseases, conditions, or disorders mediated by USP1 and / or PARP1 proteins. Accordingly, the present disclosure is generally directed to a method for treating a disease, condition, or disorder responsive to inhibition of USP1 and / or PARP1 proteins in an animal suffering from or at risk of suffering from the disorder, comprising administering to the animal an effective amount of one or more combinations of the present disclosure.

[0191] The present disclosure is further directed to a method of inhibiting USP1 and / or PARP1 protein in an animal in need thereof, comprising administering to the animal a therapeutically effective amount of a combination of the present disclosure.

[0192] In some embodiments, the combination of the present disclosure can be used to inhibit the activity of USP1 and / or PARP1 protein. For example, in some embodiments, the method of inhibiting USP1 and / or PARP1 protein comprises contacting USP1 and / or PARP1 protein with the combination of the present disclosure. The contacting can occur in vitro or in vivo.

[0193] In some embodiments, the combination of the present disclosure can be used to treat USP1 and / or PARP1 protein-mediated disorders. USP1 and / or PARP1 protein-mediated disorders are pathological conditions known to involve USP1 and / or PARP1 proteins. In some embodiments, USP1 and / or PARP1 mediated disorders are proliferative diseases such as cancer. In some embodiments, the combination of the present disclosure can delay, reduce, or prevent tumor rebound (rapid regrowth).

[0194] In some embodiments, the combinations of the present disclosure are less toxic than the PARP1-selective inhibitor alone (eg, because lower amounts of the PARP1-selective inhibitor are administered).

[0195] Provided herein are various methods of treating diseases and disorders with the disclosed combinations. Exemplary diseases and disorders that can be treated with the disclosed combinations include, but are not limited to, cancer.

[0196] In some embodiments, methods of treating cancer with the disclosed combinations are provided, including administering a therapeutically effective amount of the disclosed combination to a subject with cancer.

[0197] In some embodiments, the cancer that can be treated with the combination of the present disclosure is selected from blood cancer, lymphatic cancer, and DNA damage repair pathway defective cancer.In some embodiments, the cancer that can be treated with the combination of the present disclosure is a cancer that comprises cancer cells with a mutation in the gene encoding p53.In some embodiments, the cancer that can be treated with the combination of the present disclosure is a cancer that comprises cancer cells with a loss-of-function mutation in the gene encoding p53.In some embodiments, the cancer that can be treated with the combination of the present disclosure is a cancer that comprises cancer cells with a mutation in the gene encoding BRCA1.In some embodiments, the cancer that can be treated with the combination of the present disclosure is a cancer that comprises cancer cells with a mutation in the gene encoding BRCA2.In some embodiments, the cancer that can be treated with the combination of the present disclosure is a cancer that comprises cancer cells with a loss-of-function mutation in the gene encoding ATM.

[0198] In some embodiments, the cancer that can be treated with the combination of the present disclosure is advanced solid tumor.Advanced solid tumor is unresectable and / or metastatic solid tumor.In some embodiments, the cancer that can be treated with the combination of the present disclosure is benign or metastatic solid tumor that can be resected, and the combination is administered before and / or after resection treatment.

[0199] In some embodiments, the cancer that can be treated with the combination of the present disclosure is selected from non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer. In some embodiments, the cancer is uterine cancer. In some embodiments, the cancer is peritoneal cancer. In some embodiments, the cancer is endometrial cancer, and in some embodiments, the cancer is ovarian cancer or breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is triple-negative breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the ovarian cancer is a BRCA1 mutant cancer, a BRCA2 mutant cancer, or a p53 mutant cancer. In some embodiments, the ovarian cancer is a BRCA1 mutant cancer and a p53 mutant cancer. In some embodiments, the ovarian cancer is a BRCA1 and a BRCA2 mutant cancer. In some embodiments, the ovarian cancer is a BRCA2 mutant cancer.

[0200] In some embodiments, the cancer that can be treated with the combination of the present disclosure is selected from the group consisting of bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma and meningioma; soft tissue cancer, including rhabdoid and sarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer, uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.In some embodiments, the cancer that can be treated with the combination of the present disclosure is selected from the group consisting of uterine cancer, peritoneal cancer and endometrial cancer.

[0201] Provided herein are various methods of treating cancer with the disclosed combinations. In some embodiments, a therapeutically effective amount of the disclosed combination is administered to a subject with cancer.

[0202] In some embodiments, such methods comprise administering a therapeutically effective amount of a combination of the present disclosure to a subject with triple-negative breast cancer.

[0203] In some embodiments, the combinations of the present disclosure are used to treat cancers that are homologous recombination deficient. In some embodiments, the combinations of the present disclosure are used to treat cancers that contain cancer cells with mutations in the gene encoding p53. In some embodiments, the combinations of the present disclosure are used to treat cancers that contain cancer cells with loss-of-function mutations in the gene encoding p53. In some embodiments, the combinations of the present disclosure are used to treat cancers that do not have a defect in the homologous recombination pathway.

[0204] In some embodiments, the combinations of the present disclosure are used to treat cancers that are BRCA1 mutant cancers. In some embodiments, the combinations of the present disclosure are used to treat cancers that are BRCA2 mutant cancers. In some embodiments, the combinations of the present disclosure are used to treat cancers that are BRCA1 mutant cancers and BRCA2 mutant cancers. In some embodiments, the cancer is not a BRCA1 mutant cancer or a BRCA2 mutant cancer. In some embodiments, the cancer is a BRCA1 deficient cancer. In some embodiments, the cancer is a BRCA2 deficient cancer. In some embodiments, the cancer is a BRCA1 deficient cancer and a BRCA2 mutant cancer.

[0205] In some embodiments, the combinations of the present disclosure are used to treat cancer that is an ATM mutant cancer. In some embodiments, the cancer is not an ATM mutant cancer. In some embodiments, the cancer is an ATM deficient cancer.

[0206] In some embodiments, the combination of the present disclosure is used to treat cancer that is PARP inhibitor-resistant or refractory cancer.In some embodiments, the combination of the present disclosure is used to treat cancer that is PARP inhibitor-resistant or refractory BRCA1-deficient cancer.In some embodiments, the cancer is olaparib-resistant or refractory cancer.In some embodiments, the cancer is AZD-5305-resistant or refractory cancer.

[0207] In some embodiments, the cancer is a BRCA1 and / or BRCA2 mutant cancer comprising cells with elevated levels of RAD18, where, for example, the elevated level of RAD18 is at least as high as the RAD18 protein and / or mRNA level in ES2 cells (ES2 cells are publicly available, e.g., from the American Type Culture Collection (ATCC; CRL-1978)), or the elevated level of RAD18 is at least as high as the RAD18 protein and / or mRNA level in HEP3B217 cells (HEP3B217 cells are publicly available, e.g., from the ATCC (HB-8064)). In some embodiments, the triple-negative breast cancer is a BRCA1 and / or BRCA2 mutant cancer.

[0208] In some cases, the methods described herein include detecting RAD51 (e.g., RAD51 protein, RAD51 protein foci, and / or RAD51 mRNA) levels in cancer cells (e.g., using a sample obtained from the cancer cells). RAD51 protein levels can be detected using, for example, immunofluorescence, Western blot, fluorescence-activated cell sorting (FACS), and / or immunohistochemistry. RAD51 mRNA levels can be detected using quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR).

[0209] In some embodiments, the combinations of the present disclosure are used to treat cancers that have previously been treated with platinum therapy. In some embodiments, the combinations of the present disclosure are used to treat platinum-resistant cancers. In some embodiments, the combinations of the present disclosure are used to treat platinum-refractory cancers. In some embodiments, the platinum therapy is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, and combinations thereof.

[0210] In some cases, the cancer is a solid cancer. In some cases, the cancer is a hematological / lymphatic cancer. In some cases, the cancer is a DNA damage repair pathway deficient cancer. In some cases, the cancer is a homologous recombination deficient cancer. In some cases, the cancer comprises cancer cells with a mutation in a gene encoding p53. In some cases, the cancer comprises cancer cells with a loss-of-function mutation in a gene encoding p53. In some cases, the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer (including triple-negative breast cancer). In some cases, the cancer is ovarian cancer or breast cancer (including triple-negative breast cancer). In some cases, the cancer is ovarian cancer. In some cases, the cancer is breast cancer (including triple-negative breast cancer). In some cases, the cancer is uterine cancer. In some cases, the cancer is peritoneal cancer. In some cases, the cancer is endometrial cancer.

[0211] In some embodiments, the combinations of the present disclosure are used in combination with one or more additional therapeutic agents to treat cancer.

[0212] In some embodiments, provided herein are combinations of the present disclosure for use as a medicament or for use in the manufacture of a medicament, e.g., for the treatment of cancer. In some embodiments, provided herein are combinations of the present disclosure for use in a method of treating cancer.

[0213] Pharmaceutical Combination Composition The combinations of the present disclosure can be administered to a mammal in the form of raw chemicals, free of any other components, or they can also be administered to a mammal as part of a pharmaceutical composition comprising the compounds in combination with a suitable pharmaceutically acceptable carrier (see, e.g., Gennaro, Remington: The Science and Practice of Pharmacy with Facts and Comparisons: Drugfacts Plus, 20th ed. (2003); Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ed., Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook of Pharmaceutical Excipients, 3rd ed., Pharmaceutical Press (2000)). Such carriers can be selected from pharmaceutically acceptable excipients and adjuvants. The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable vehicle" encompasses any of the standard pharmaceutical carriers, solvents, surfactants, or vehicles. Standard pharmaceutical carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 19th ed. 1995. The USP1 inhibitor and the PARP1 selective inhibitor can be present in the same pharmaceutical composition; or, the USP1 inhibitor and the PARP1 selective inhibitor can be present in separate pharmaceutical compositions that can be administered simultaneously or sequentially.

[0214] Pharmaceutical combination compositions of the present disclosure may be prepared as liquid suspensions or solutions using a liquid, such as oil, water, alcohol, and combinations thereof.

[0215] As provided herein, a USP1 inhibitor (e.g., 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine or a pharmaceutically acceptable salt, solvate, hydrate, or cocrystal thereof) can be present in a pharmaceutical composition, including any composition in which such an inhibitor is combined with one or more pharmaceutically acceptable carriers. In some embodiments, the USP1 inhibitor is present in the composition in an amount effective to achieve its intended therapeutic purpose.

[0216] In some embodiments, the disclosure provides a pharmaceutical composition comprising a USP1 inhibitor (e.g., 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine or a pharmaceutically acceptable salt, solvate, hydrate, or cocrystal thereof) and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises crystalline Form 2 of the gentisic acid cocrystal of Formula I.

[0217] Pharmaceutical combination compositions to be used for in vivo administration can be sterile, which is readily accomplished, for example, by filtration through sterile filtration membranes.

[0218] Pharmaceutical combination compositions within the scope of the present disclosure include any composition in which the USP1 inhibitor and PARP1 selective inhibitor of the present disclosure are combined with one or more pharmaceutically acceptable carriers. In one embodiment, the USP1 inhibitor and PARP1 selective inhibitor of the present disclosure are present in the composition in an amount effective to achieve their intended therapeutic purpose.

[0219] The pharmaceutical combination composition of the present disclosure can be administered to patients who can experience the beneficial effects of the combination of the present disclosure.The most important of such patients are mammals, for example, humans and companion animals, but the present disclosure is not intended to be so limited.In one embodiment, the patient is human.In another embodiment, the pharmaceutical combination composition of the present disclosure can be administered to patients with PARP inhibitor-resistant or refractory cancer.In another embodiment, the pharmaceutical combination composition of the present disclosure can be administered to patients with PARP inhibitor-resistant or refractory BRCA1-deficient cancer.

[0220] In another embodiment, the present disclosure provides a kit comprising the combination of the present disclosure, packaged in a manner that facilitates its use for carrying out the method of the present disclosure.In one embodiment, the kit comprises the USP1 inhibitor and PARP1 selective inhibitor of the present disclosure packaged in a container such as a sealed bottle or container, and a label that describes the use of the compound to carry out the method of the present disclosure is attached to the container or included in the kit.In one embodiment, the combination composition is packaged in a unit dosage form.The kit can further comprise a device suitable for administering the combination composition according to the intended administration route.In some embodiments, the present disclosure provides a kit comprising the USP1 inhibitor and PARP1 selective inhibitor of the present disclosure, or their pharmaceutically acceptable salts or solvates, and instructions for administering the compound or their pharmaceutically acceptable salts or solvates to cancer patients.

[0221] In some embodiments, the present disclosure provides a pharmaceutical combination composition comprising a USP1 inhibitor and a PARP1 selective inhibitor of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0222] In some embodiments, the present disclosure provides a pharmaceutical combination composition comprising a USP1 inhibitor and a PARP1 selective inhibitor of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the combination binds to a protein encoded by the USP1 gene and / or the PARP1 gene.

[0223] In some embodiments, the present disclosure provides a pharmaceutical combination composition comprising a USP1 inhibitor and a PARP1 selective inhibitor of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is for use in treating cancer.

[0224] In some embodiments, the present disclosure provides a pharmaceutical combination composition comprising a USP1 inhibitor and a PARP1 selective inhibitor of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is for the manufacture of a medicament for treating cancer. [Example]

[0225] Example 1: Antitumor activity of Formula I cocrystals in combination with the selective PARP1 inhibitor AZD5305 in a patient-derived breast xenograft model in nude mice The USP1 inhibitor used in this example is the co-crystal described in paragraphs

[0140] and

[0143] to

[0155] above. This co-crystal can be prepared as described in Example 6 of published PCT Publication WO 2022 / 094096. The Formula I co-crystal is a co-crystal of gentisic acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

[0226] The antitumor activity of the USP1 inhibitor cocrystal of Formula I in combination with AZD5305 was evaluated in mice using an olaparib-resistant patient-derived breast xenograft model in nude mice, as shown in Figures 1A-1C. Six- to nine-week-old female athymic nude mice from Envigo were anesthetized and injected with 20 mm 3 Tumor fragments were placed subcutaneously through a flank incision. 3 When tumor volumes in the range of 0.1 to 1.0 mmHg were reached, mice were randomized into groups of 10 and assigned to the following groups: control, Formula I (100 mg / kg), AZD5305 (10 mg / kg), or a combination of Formula I (100 mg / kg) and AZD5305 (10 mg / kg). Compounds were administered by oral gavage once daily for at least 42 days depending on the treatment group. Body weight and tumor volume were measured twice weekly. Tumor volumes were calculated as the mean and standard error of the mean for each treatment group.

[0227] The data in Figure 1A show that the combination treatment group demonstrated enhanced antitumor activity in a patient-derived subcutaneous mouse model compared to equivalent doses of either Formula I or AZD5305 alone.

[0228] To assess combination activity in mice bearing tumors resistant to PARP inhibitors, five mice that were resistant to AZD5305 monotherapy were then co-administered with Formula I (100 mg / kg) and AZD5305 (10 mg / kg) on ​​day 49 of treatment. Compared to the remaining animals treated with AZD5305 monotherapy, the addition of Formula I resulted in a reduction in tumor volume, as shown in Figure 1B.

[0229] The tolerability of AZD5305 (10 mg / kg) and Formula I (100 mg / kg) in the combination efficacy study was assessed by monitoring body weight and calculating the % weight change from the weight on the day treatment began (day 0), as shown in Figure 1C.

[0230] Example 2: In vitro efficacy In vitro experiments were performed using a colony-forming unit (CFU) assay. UWB1.289 parental cells or UWB1.289BRCA1-overexpressing cells were plated at a concentration of 2500 cells / well on day -1. On day 0, wells were treated with DMSO or increasing concentrations of Formula I and / or AZD5305. On day 10, when clearly dispersed colonies were observed in DMSO-treated wells, the cells were fixed and stained with 0.1% crystal violet in 10% ethanol for 20 minutes at room temperature. The plates were imaged, and the amount of crystal violet staining in each well was quantified by extracting the crystal violet in 10% acetic acid and measuring the absorbance at 565 nm. The CFU results are shown in Figures 4 and 5. These results demonstrate that a USP1 inhibitor and a PARP1-selective inhibitor are an effective combination in UWB1.289 and UWB1.289BRCA1-overexpressing cells.

[0231] The present invention having now been fully described, it will be understood by those skilled in the art that the same can be practiced within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any of its aspects.

[0232] Other aspects of the present invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.

[0233] All patents and publications cited herein are hereby incorporated by reference in their entirety.

Claims

1. 1. A method of treating cancer in a subject, comprising administering to the subject (i) an inhibitor of ubiquitin-specific processing protease 1 (USP1) and (ii) a selective inhibitor of poly ADP-ribose polymerase 1 (PARP1), or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof; USP1 inhibitors, (a) Formula I: 【Chemical 1】 (b) Formula II: 【Chemistry 2】 (c) Formula III: 【Chemistry 3】 and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof, method.

2. 1. A method of delaying, reducing, or preventing tumor rebound in a subject, comprising administering to the subject (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor and (ii) a PARP1 selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof; USP1 inhibitors, (a) Formula I: 【Chemistry 4】 (b) Formula II: 【Chemistry 5】 (c) Formula III: 【Chemistry 6】 and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof, method.

3. 3. The method of claim 1 or 2, wherein the PARP1 selective inhibitor is AZD5305 or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid or polymorph thereof.

4. The method of any one of claims 1 to 3, wherein the USP1 inhibitor is a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

5. 4. The method of any one of claims 1 to 3, wherein the USP1 inhibitor is a compound of formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

6. 4. The method of any one of claims 1 to 3, wherein the USP1 inhibitor is a compound of formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

7. The method of any one of claims 1 to 6, wherein the USP1 inhibitor is provided as a co-crystal of the pharmaceutically acceptable acid and the USP1 inhibitor.

8. 8. The method of claim 7, wherein the USP1 inhibitor is a co-crystal of a pharmaceutically acceptable acid and 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, wherein the pharmaceutically acceptable acid can be a gentisic acid, benzoic acid, or salicylic acid co-crystal.

9. 9. The method of any one of claims 1 to 8, wherein administration of the USP1 inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, and the PARP1 selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, results in a synergistic effect.

10. 10. The method of any one of claims 1 to 9, wherein the method is administered in a therapeutically effective amount sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) a reduction in tumor size, (ii) an increase in the rate of tumor regression, and (iii) a reduction or inhibition of tumor growth.

11. 11. The method of any one of claims 1 to 10, wherein the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof delays, reduces, or prevents tumor rebound.

12. 12. The method of any one of claims 1 to 11, wherein the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, are administered sequentially.

13. 12. The method of any one of claims 1 to 11, wherein the USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof and the PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof are administered simultaneously.

14. The method according to any one of claims 1 to 13, wherein the USP1 inhibitor and / or the PARP1 selective inhibitor is administered at a dose that is not effective as a single agent.

15. The method of any one of claims 1 to 14, wherein the subject is a mammal, which may be a human.

16. 1. A combination composition comprising (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor, and (ii) a PARP1 selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof, wherein the USP1 inhibitor is: (a) Formula I: 【Chemistry 7】 (b) Formula II: 【Chemistry 8】 (c) Formula III: 【Chemistry 9】 and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids or polymorphs thereof.

17. 17. The composition of claim 16, wherein the PARP1 selective inhibitor is AZD5305 and its pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals.

18. 18. The composition of claim 16 or 17, wherein the USP1 inhibitor is a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid or polymorph thereof.

19. 18. The composition of claim 16 or 17, wherein the USP1 inhibitor is a compound of formula II or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid or polymorph thereof.

20. 18. The composition of claim 16 or 17, wherein the USP1 inhibitor is a compound of formula III or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof:

21. The composition of any one of claims 16 to 20, wherein the USP1 inhibitor is provided as a co-crystal of a pharmaceutically acceptable acid and the USP1 inhibitor, which may be a co-crystal of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, wherein the co-crystal is a gentisic acid, benzoic acid, or salicylic acid co-crystal.

22. The composition according to any one of claims 16 to 21, wherein the USP1 inhibitor and the PARP1 selective inhibitor are for simultaneous administration.

23. The composition according to any one of claims 16 to 21, wherein the USP1 inhibitor and the PARP1 inhibitor are for sequential administration.

24. Use of the composition according to any one of claims 16 to 23 for the manufacture of a medicament for the treatment of cancer.

25. A pharmaceutical composition comprising the composition of any one of claims 16 to 23 and a pharmaceutically acceptable carrier.

26. 26. The pharmaceutical composition of claim 25 for use in the treatment of cancer.

27. A kit comprising the composition of any one of claims 16 to 23 or the pharmaceutical composition of claim 25 or 26, and instructions for administering the combination to a subject with cancer.

28. The method according to any one of claims 1 to 15, the use according to claim 24, the pharmaceutical composition according to claim 26 or the kit according to claim 27, wherein the cancer is an advanced solid tumor.

29. The method according to any one of claims 1 to 15, the use according to claim 24, the pharmaceutical composition according to claim 26, or the kit according to claim 27, wherein the cancer is selected from the group consisting of blood cancer, lymphatic cancer, DNA damage repair pathway deficient cancer, homologous recombination deficient cancer, cancer comprising cancer cells having a mutation in the gene encoding p53, and cancer comprising cancer cells having a loss-of-function mutation in the gene encoding p53.

30. 30. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 29, wherein the cancer is selected from the group consisting of brain cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, skin cancer, uterine cancer, peritoneal cancer, and endometrial cancer, and breast cancer.

31. The method, use, pharmaceutical composition or kit according to any one of claims 1 to 15, 24, or 26 to 29, wherein the cancer is ovarian cancer or breast cancer.

32. The method, use, pharmaceutical composition or kit according to any one of claims 1 to 15, 24, or 26 to 29, wherein the cancer is ovarian cancer.

33. 30. The method, use, pharmaceutical composition or kit according to any one of 1 to 15, 24, or 26 to 29, wherein the cancer is breast cancer.

34. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 29, wherein the cancer is triple-negative breast cancer.

35. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 34, wherein the cancer is a DNA damage repair pathway deficient cancer.

36. 36. The method, use, pharmaceutical composition or kit of claim 35, wherein the cancer is a homologous recombination deficient cancer.

37. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 36, wherein the cancer is a BRCA1 mutant cancer.

38. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 37, wherein the cancer is a BRCA2 mutant cancer.

39. The method, use, pharmaceutical composition or kit according to any one of 1 to 15, 24, or 26 to 38, wherein the cancer is a BRCA1 mutant cancer and a BRCA2 mutant cancer.

40. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 39, wherein the cancer is a PARP inhibitor-resistant or refractory cancer, an olaparib-resistant or refractory cancer, or an AZD-5305-resistant or refractory cancer.

41. 41. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 40, wherein the cancer comprises cancer cells having a mutation in the gene encoding ATM.

42. 42. The method, use, pharmaceutical composition or kit of any one of claims 1 to 15, 24, or 26 to 41, wherein the cancer has previously been treated with platinum therapy, and the cancer may be platinum-resistant or platinum-refractory.

43. 43. The method, use, pharmaceutical composition or kit of claim 42, wherein the platinum therapy is selected from the group consisting of cisplatin, carboplatin, oxaliplatin and combinations thereof.

44. A method for treating a USP1 protein-mediated disorder and / or a PARP1 protein-mediated disorder, comprising administering to a subject in need of said treatment a composition according to any one of claims 16 to 23 or a pharmaceutical composition according to claims 25, 26, or 28 to 44 in an amount effective for treating a USP1 protein-mediated disorder and / or a PARP1 protein-mediated disorder.

45. A method for inhibiting USP1 protein and / or PARP1 protein, comprising contacting the USP1 protein and / or PARP1 protein with a composition according to any one of claims 16 to 23 or a pharmaceutical composition according to claims 25, 26, or 28 to 44.

46. 46. ​​The method of claim 45, wherein the contacting occurs in vitro.

47. 46. ​​The method of claim 45, wherein the contacting occurs in vivo.

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