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

Abstract

The present disclosure provides a therapeutic combination comprising (i) an 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 comprises # imgabs0 # or a pharmaceutically acceptable salt, a hydrate, a solvate, an amorphous solid, a polymorphic substance or a co-crystal thereof. The present disclosure also relates to the use of said combination in the inhibition of USP1 and / or PARP1 protein and / or in the treatment of diseases responsive to the inhibition of USP1 and / or PARP1P protein and USP1 and / or PARP1 activity. The combinations of the present disclosure are particularly useful for the treatment of cancer.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 376,987 filed on September 23, 2022, U.S. Provisional Patent Application No. 63 / 380,742 filed on October 24, 2022, and U.S. Provisional Patent Application No. 63 / 496,289 filed on April 14, 2023, all of which are incorporated herein by reference in their entireties. Reference to electronically submitted sequence listings

[0002] The contents of the electronically submitted Sequence Listing (Name: 4195_034PC03_Seqlisting_ST26; Size: 2,057 bytes; Creation Date: September 21, 2023) are hereby incorporated by reference in their entirety. Technical Field

[0003] The present disclosure provides therapeutic combinations of ubiquitin-specific processing protease 1 (USP1) inhibitors and selective poly (ADP-ribose) polymerase 1 (PARP1) inhibitors. Also provided are methods of treating cancer comprising administering the combination. Background Art

[0004] Ubiquitin is a small protein (76 amino acids) that is attached to target proteins post-transcriptionally. The outcome of ubiquitination depends on the number and linkage topology of the ubiquitin molecules conjugated to the target protein. For example, proteins displaying polyubiquitin chains linked to lysine 48 are often targets for proteasomal degradation, while monoubiquitination or polyubiquitin chains linked through other lysines regulate non-proteolytic functions such as cell cycle regulation, DNA damage repair, transcription, and endocytosis. Ubiquitination is a reversible process in which enzymes called deubiquitinases remove ubiquitin from target proteins.

[0005] USP1 is a deubiquitinase that plays a role 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), two proteins that play important roles in the translesion synthesis (TLS) and Fanconi anemia (FA) pathways, respectively. The USP1 / UAF1 complex also deubiquitinates Fanconi anemia complementation group 1 (FANCI). These two pathways are crucial for repairing DNA damage caused by DNA cross-linking agents such as cisplatin and mitomycin C (MMC).

[0006] The poly(ADP-ribose) polymerase (PARP) family of enzymes plays a role in DNA repair and genomic integrity. PARP is essential for single-strand break repair and base excision repair pathways. A key enzymatic activity is the addition of ADP-ribose to substrate proteins via the cleavage of NAD+ and the release of nicotinamide. This poly(ADP-ribosylation) ("PARylation") activity is activated by DNA strand breaks, resulting in the addition of Par to PARP itself and other DNA repair enzymes. PARP is crucial for the recruitment of DNA repair proteins to sites of damage.

[0007] Homologous recombination is a DNA repair process crucial for accurate DNA damage repair. BRCA1 / 2, along with other Fanconi anemia pathway genes (e.g., RAD51D, NBN, ATM), are integral to homologous recombination-mediated DNA repair. Mutations in genes encoding homologous recombination factors play a role in the progression of certain cancers. PARP inhibitors prevent the repair of single-strand breaks (SSBs) and promote their conversion to double-strand breaks (DSBs), resulting in synthetic lethality in cancer cells lacking proficient DSB machinery such as homologous recombination.

[0008] There remains an unmet medical need for more effective therapies, such as combination therapies, to treat cancer. Summary of the Invention

[0009] Provided herein are combinations of (i) a ubiquitin-specific processing protease 1 (USP1) inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal 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 co-crystal thereof. Also provided herein are methods of using such combinations to treat subjects with cancer.

[0010] In one aspect, the present disclosure relates to a combined 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 compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs, or co-crystals thereof. In another aspect, Formula I, Formula II, or Formula III is provided as a co-crystal. In some aspects, Formula I, Formula II, or Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0011] In some aspects, 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 aspect, 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 aspects, Formula I is provided as a co-crystal with a pharmaceutically acceptable acid.

[0013] In another aspect, 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 aspects, Formula II is provided as a co-crystal with a pharmaceutically acceptable acid.

[0014] In another aspect, 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 aspects, Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

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

[0016] In another aspect, 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 aspect, 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 aspect, 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 the use of the combinatorial composition in the manufacture of a medicament for the treatment of cancer.

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

[0021] In one aspect, the pharmaceutical composition is used to treat cancer.

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

[0023] In another aspect, the present disclosure relates to 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 cocrystal thereof, wherein the USP1 inhibitor is a compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs, or co-crystals thereof. In another aspect, Formula I, Formula II, or Formula III is provided as a co-crystal. In some aspects, Formula I, Formula II, or Formula III is provided as a co-crystal with a pharmaceutically acceptable acid.

[0024] In one aspect 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 aspect of the method, the USP1 inhibitor is a compound of Formula I, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal thereof. In some aspects, Formula I is provided as a cocrystal with a pharmaceutically acceptable acid.

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

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

[0028] In another aspect, 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 aspect, 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 aspect, 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 aspect 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 aspect of the present disclosure, a USP1 inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, and a PARP1-selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, is administered in a therapeutically effective amount sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) reducing tumor size, (ii) increasing the rate of cancer tumor regression, (iii) reducing or inhibiting cancer tumor growth, and (iv) reducing the toxicity of the PARP1-selective inhibitor administered as a monotherapy (e.g., caused by administering less PARP1-selective inhibitor). In one aspect of the present disclosure, a USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, and a PARP1-selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof is administered in a therapeutically effective amount sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) reducing tumor size, (ii) increasing the rate of cancer tumor regression, and (iii) reducing or inhibiting cancer tumor growth. In one aspect of the present disclosure, a USP1 inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, and a PARP1-selective inhibitor or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof is administered in an amount sufficient to reduce the toxicity of the PARP1-selective inhibitor administered as a monotherapy (e.g., caused by administering less PARP1-selective inhibitor).

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

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

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

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

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

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

[0039] In some aspects, 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, endometrial cancer, and breast cancer.

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

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

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

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

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

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

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

[0047] In some aspects, 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 carcinoma 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 aspects, the cancer is a cancer that is defective in a DNA damage repair pathway.

[0049] In some aspects, the cancer is a BRCA1 mutant cancer. In some aspects, the BRCA1 mutation is a germline mutation. In some aspects, the BRCA1 mutation is a somatic mutation. In some aspects, the BRCA1 mutation results in BRCA1 deficiency.

[0050] In some aspects, the cancer is a BRCA2 mutant cancer. In some aspects, the BRCA2 mutation is a germline mutation. In some aspects, the BRCA2 mutation is a somatic mutation. In some aspects, the BRCA2 mutation results in BRCA2 deficiency.

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

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

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

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

[0055] In some aspects, the cancer is a PARP inhibitor-refractory or resistant cancer. In some aspects, the cancer is a PARP inhibitor-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 mutant cancer. In some aspects, the cancer is a PARP inhibitor-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 deficient cancer.

[0056] In some aspects, the cancer is an olaparib-refractory or resistant cancer. In some aspects, the cancer is an olaparib-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 mutant cancer. In some aspects, the cancer is an olaparib-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 deficient cancer.

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

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

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

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

[0061] In some aspects, the cancer has been previously treated with platinum therapy. In some aspects, the cancer is platinum-resistant or platinum-refractory. In some aspects, 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 disease mediated by a USP1 protein and / or a disease mediated by a PARP protein, comprising administering to a subject in need thereof an effective amount of a USP1 inhibitor of Formula I, Formula II or Formula III, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, and a PARP1-selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or cocrystal thereof, to treat the disease mediated by a USP1 protein and / or the disease mediated by a PARP protein.

[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 aspects, the contacting occurs in vitro.

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

[0066] In one aspect 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 aspect of the method, the USP1 inhibitor is a compound of Formula I, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or cocrystal thereof. In some aspects, Formula I is provided as a cocrystal with a pharmaceutically acceptable acid.

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

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

[0070] In another aspect, 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] On the other hand, 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 aspect, 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 in 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 aspect, the pharmaceutical composition is used to treat cancer.

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

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

[0078] It is to be understood that both the foregoing summary and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1A It was shown that the USP1 inhibitor of Formula I enhanced the anti-tumor activity of AZD5305 in a breast PDX model.

[0080] Figure 1B The efficacy of the combination of a USP1 inhibitor of Formula I and AZD5305 in mice with PARP inhibitor-resistant tumors is shown.

[0081] Figure 1C The tolerability of the combination of the USP1 inhibitor of formula I and AZD5305 is shown.

[0082] Figure 2 The XRPD pattern corresponding to crystalline State 2 is shown.

[0083] Figure 3 The DSC and TGA thermograms corresponding to crystalline State 2 are shown.

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

[0085] Figure 5 Shown are colony forming assays (CFU) of UWB1.289 BRCA1 overexpressing cells treated with Formula I and / or AZD5305. DETAILED DESCRIPTION

[0086] One aspect of the present disclosure is based on the combined use of an inhibitor of the ubiquitin-specific processing protease 1 (USP1) protein and a PARP1-selective inhibitor. The combination can be used to inhibit USP1 protein and / or PARP protein and to treat diseases, disorders, or conditions responsive to inhibition of USP1 protein and / or PARP protein, such as cancer.

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

[0088] In some aspects, the therapeutically effective amount of the USP1 inhibitor and the PARP1-selective inhibitor is sufficient to produce a therapeutic effect, including: (i) reducing tumor size, (ii) increasing the rate of cancer tumor regression, (iii) reducing or inhibiting cancer tumor growth, and / or (iv) reducing the toxicity of the PARP1-selective inhibitor administered as a monotherapy (e.g., caused by administering less PARP1-selective inhibitor). In some aspects, the USP1 inhibitor and the 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 of the PARP inhibitor olaparib were poorly tolerated. See, for example, Samol, J., et al., Invest. New Drugs, 30:1493-500 (2012) ("Further development of the combination of olaparib and topotecan was not explored due to dose-limiting hematologic adverse events and the resulting subtherapeutic MTD."). definition

[0090] Unless otherwise defined, all scientific and technological terms used herein have the same meaning as those of ordinary skill in the art to which the present disclosure pertains. In the event of any conflict, the present application (including definitions) shall prevail. Unless the context otherwise requires, singular terms shall include plural terms and plural terms shall also include singular terms. 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 were specifically and individually indicated to be incorporated by reference.

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

[0092] To further define the present disclosure, the following terms and definitions are provided.

[0093] It should be understood that aspects described herein include "consisting of" and / or "consisting essentially of" aspects. As used herein, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated. The term "or" as used herein does not imply that alternatives are mutually exclusive.

[0094] In this application, unless clearly stated or understood by one skilled in the art, the use of "or" means "and / or." In the context of multiple dependent claims, the use of "or" refers to more than one preceding independent or dependent claim.

[0095] As used herein, the term "and / or" should be understood as specific disclosure of each of the two specified features or components, whether or not the other is present. Thus, the term "and / or" as used in the phrase "A and / or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to cover each of the following: 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 (alone); B (alone); and C (alone).

[0096] As used herein, the term "about" includes the number ± 10%. Thus, "about 10" means 9 to 11. One skilled in the art will understand that references herein to "about" a value or parameter include (and describe) the value or parameter itself. For example, a description referring to "about X" includes a description of "X."

[0097] The present disclosure covers 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, etc.; alkaline earth metal salts such as calcium salts, magnesium salts, etc.; organic amine salts such as triethylamine salts, pyridinium salts, pyridinium salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, N,N'dibenzylethylenediamine salts, etc.; inorganic acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, etc.; organic acid salts such as citrates, lactates, tartrates, maleates, fumarates, mandelates, acetates, dichloroacetates, trifluoroacetates, oxalates, formates, etc.; sulfonates such as methanesulfonates, benzenesulfonates, p-toluenesulfonates, etc.; and amino acid salts such as arginine salts, aspartates, glutamates, etc. As used herein, the term "pharmaceutically acceptable salt" refers to any salt of a USP1 inhibitor or PARP1-selective inhibitor of the present disclosure, obtained, for example, by reaction with an acid or base, that is physiologically tolerated in the target patient (eg, a mammal, such as a human).

[0098] Acid addition salts can be formed by mixing a solution of a specific 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. Basic salts can be formed by mixing a solution of a USP1 inhibitor or PARP1-selective inhibitor of the present disclosure with a solution of a pharmaceutically acceptable non-toxic base, such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, etc.

[0099] In some aspects 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 aspects, a pharmaceutically acceptable salt of a compound of Formula I, Formula II, or Formula III is a crystalline form.

[0100] In some aspects, the pharmaceutically acceptable acid is selected from the group consisting of: 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, caproic acid, hippuric acid, 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, ethyl gallate, 4-hydroxybenzoic acid, methyl 4-hydroxybenzoate, 3,4,5-trihydroxybenzoic acid, niacinamide, L-proline, and D-sorbitol. In some aspects, the pharmaceutically acceptable acid is selected from the group consisting of: 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, caproic acid, hippuric acid, hydrobromic acid, hydrochloric acid, lactic acid, maleic acid In some aspects, 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 aspects, 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 aspects, the pharmaceutically acceptable acid is selected from the group consisting of: hydrochloric acid, hydrobromic acid, ethanedisulfonic acid, methanesulfonic acid and gentisic acid. In some aspects, the pharmaceutically acceptable acid is a benzoic acid derivative. In some aspects, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with one or more hydroxyl groups. In some aspects, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with one hydroxyl group. In some aspects, the benzoic acid derivative substituted with one hydroxyl group is salicylic acid. In some aspects, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with two hydroxyl groups. In some aspects, the benzoic acid derivative substituted with two hydroxyl groups is gentisic acid. In some aspects, the pharmaceutically acceptable acid is a benzoic acid derivative substituted with three hydroxyl groups. In some aspects, the benzoic acid derivative substituted with three hydroxyl groups is gallic acid. In some aspects, the benzoic acid derivative is selected from the group consisting of salicylic acid, gentisic acid, and gallic acid. In some aspects, the pharmaceutically acceptable acid is hydrochloric acid. In some aspects, the pharmaceutically acceptable acid is hydrobromic acid. In some aspects, the pharmaceutically acceptable acid is ethanedisulfonic acid.In some aspects, the pharmaceutically acceptable acid is methanesulfonic acid. In some aspects, the pharmaceutically acceptable acid is gentisic acid. In some aspects, the pharmaceutically acceptable acid is benzoic acid. In some aspects, the pharmaceutically acceptable acid is salicylic acid. In some aspects, the pharmaceutically acceptable acid is gallic acid.

[0101] The present disclosure includes the preparation and use of solvates of USP1 inhibitors and / or PARP1-selective inhibitors. Solvates generally do not significantly alter the physiological activity or toxicity of the compound and therefore can act as pharmacological equivalents. The term "solvate" as used herein is a combination, physical association and / or solvation of a USP1 inhibitor or PARP1-selective inhibitor of the present disclosure with solvent molecules, such as a disolvate, a monosolvate or a hemisolvate, wherein the ratio of solvent molecules to the compound of the present disclosure is about 2:1, about 1:1 or about 1:2, respectively. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain cases, the solvate can be isolated, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Therefore, "solvate" includes both solution phases and separable solvates. The USP1 inhibitors or PARP1-selective inhibitors of the present disclosure may exist in a solvated form with a pharmaceutically acceptable solvent (e.g., water, methanol, ethanol, etc.), and the present disclosure is intended to include both solvated and unsolvated forms of the USP1 inhibitors and / or PARP1-selective inhibitors of the present disclosure. One solvate is a hydrate. "Hydrate" refers to a specific subclass of solvates in which the solvent molecule is water. When applied to a compound, "anhydrate" refers to a solid state in which the crystal lattice of the compound does not contain structural water. Solvates can generally act pharmacologically equivalently. The 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 a solvate of fluconazole with ethyl acetate and water. E.C. van Tender et al. AAPS Pharm. Sci. Tech., 5(1): Article 12 (2004), and A.L. Bingham et al. Chem. Commun. 603-604 (2001) describe similar preparations of solvates, hemisolvates, hydrates, and the like. A typical, non-limiting method for preparing a solvate comprises dissolving a USP1 inhibitor or PARP1-selective inhibitor of the present disclosure in a desired solvent (organic solvent, water, or a mixture thereof) at a temperature of from 20° C. to about 25° C., followed by cooling the solution at a rate sufficient to form crystals, and isolating the crystals by known methods (e.g., filtration). The presence of solvent in the solvate crystals can be confirmed using analytical techniques such as infrared spectroscopy.

[0102] In some aspects of the present disclosure, the USP1 inhibitor and / or PARP1-selective inhibitor is deuterated. In some aspects, the USP1 inhibitor and / or PARP1-selective inhibitor is partially or fully deuterated, ie, one or more hydrogen atoms are replaced by a deuterium atom.

[0103] As used herein, "treatment" is a method for obtaining a beneficial or desired clinical outcome. As used herein, "treatment" encompasses any administration or application of a therapeutic agent for a disease in a mammal (including humans). For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, any one or more of the following: alleviating one or more symptoms, reducing the extent of the disease, preventing or delaying disease spread (e.g., metastasis), preventing or delaying disease recurrence, delaying or slowing disease progression, improving the disease state, inhibiting disease or disease progression, inhibiting or slowing disease or its progress, preventing its progress, and alleviating (whether partially or completely). "Treatment" also includes reducing the pathological consequences of proliferative diseases. The methods provided herein take into account any one or more of these therapeutic aspects. Consistent with the above, the term treatment does not require 100% removal of all aspects of the disease.

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

[0105] As used herein, "delay" means to postpone, hinder, slow, delay, stabilize, inhibit and / or postpone the development or progression of a disease (e.g., cancer). The length of such delay may vary, depending on the medical history and / or the individual being treated.

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

[0107] As used herein, the terms "combination," "therapeutic combination," "combination composition," "combination therapy," or "pharmaceutical combination" may include a fixed combination in the form of one dosage unit, separate dosage units, or a kit of parts or instructions for combined administration, wherein the USP1 inhibitor and the PARP1-selective inhibitor may be administered independently at the same time or separately within time intervals. The combination pharmaceutical composition may be suitable for simultaneous, separate, or sequential administration.

[0108] Combination therapies may provide and be demonstrated to be "synergistic," i.e., the effect achieved when the active ingredients are used together is greater than the sum of the effects produced by the compounds alone. A synergistic effect may include a significant reduction in the effective dose of the combination of the two active ingredients compared to the effective dose of each active ingredient when administered alone. A synergistic effect may also include a reduction in the toxicity of the combination of the two active ingredients compared to the toxicity of each active ingredient when administered alone. A synergistic effect may also be an effect that cannot be achieved by administering any of the active ingredients as a single agent. A synergistic effect may include, but is not limited to, an effect in treating cancer by reducing tumor size, inhibiting tumor growth, or increasing the survival rate of a subject. A synergistic effect may also include reducing cancer cell viability, inducing cancer cell death, and inhibiting or delaying cancer cell growth. For example, a synergistic effect may be achieved when the active ingredients are (1) formulated and administered or delivered simultaneously in a combined unit dosage formulation; (2) delivered continuously, alternatingly, or in parallel in separate formulations; or (3) delivered by other regimens. When delivered in alternation therapy, a synergistic effect may be achieved when the compounds are administered or delivered sequentially.

[0109] Determination of a synergistic interaction between a USP1 inhibitor and a PARP1-selective inhibitor can be based on the results obtained from the assays described herein. For example, the Bliss independence model can be used to assess the combination effect. The Bliss score quantifies the degree of synergy of the single agents, with a Bliss score > 0 indicating greater than simple additivity. In some aspects, a Bliss score greater than 10 indicates strong synergy. In some aspects, a score of 6 or greater indicates synergy. In some aspects, 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, such as loss of heterozygosity, telomeric allelic imbalance, and large-scale state transitions.

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

[0112] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a pharmaceutical preparation that is in such form that the biological activity of the active ingredient is effective and that contains no additional ingredients that are unacceptably toxic to a subject to which the preparation is administered. Such preparations may be sterile.

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

[0114] A "pharmaceutically acceptable carrier" refers to a conventional non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation adjuvant, or vehicle used in the art with therapeutic agents, which together comprise the "pharmaceutical composition" to be administered to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dosage and concentration employed and is compatible with the other ingredients of the formulation. A pharmaceutically acceptable carrier is suitable for the formulation employed.

[0115] A "sterile" preparation is sterile or substantially free of viable microorganisms and their spores.

[0116] The term "container" means any receptacle and closure suitable for storing, shipping, dispensing and / or handling pharmaceutical products.

[0117] The term "insert" or "package insert" refers to the information that accompanies a drug and provides instructions on how to administer the product, as well as safety and effectiveness data necessary for doctors, pharmacists, and patients to make informed decisions about the product's use. The package insert is generally considered the drug's "label."

[0118] As used herein, the term "disease" or "condition" or "disorder" refers to a condition for which treatment is needed and / or desired and represents a disturbance and / or abnormality of function that is generally considered a pathological condition or function and that may manifest itself in the form of specific signs, symptoms, and / or dysfunction. As shown 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 provides a benefit.

[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. This definition encompasses full-length proteins and fragments thereof. These terms also include post-expression modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, and the like. In addition, for the purposes of this disclosure, "polypeptide" refers to a protein including modifications (e.g., deletions, additions, and substitutions (usually of a conservative nature)) to the native sequence, as long as the protein maintains the desired activity. These modifications may be intentional, such as by site-directed mutagenesis, or may be accidental, such as by mutations in the host producing the protein or errors due to PCR amplification.

[0120] As used herein, "USP1" and "ubiquitin-specific processing protease 1" refer to any native polypeptide or USP1 encoding polynucleotide. The term "USP1" encompasses "full-length," unprocessed USP1 polypeptides as well as any form of USP1 resulting from processing within the cell (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 a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. Human USP1 sequences are known, including, for example, the sequence publicly available as UniProt No. 094782 (including isoforms). As used herein, the term "human USP1 protein" refers to a USP1 protein comprising the amino acid sequence shown in SEQ ID NO: 1:

[0121] USP1 is a deubiquitinating enzyme that functions 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. This family includes enzymes that have the ability to catalyze the transfer of ADP-ribose to target proteins (poly ADP-ribosylation). The PARP family has at least 18 members, encoded by different genes, and share homology in the conserved catalytic domain, including PARP1, PARP2, and PARP3.

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

[0124] The term "specific binding" is well known 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 binds to a particular protein or protein domain more frequently, more rapidly, for a longer period of time, and / or with greater affinity than it reacts or binds to other proteins or domains. It will 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, but not necessarily, reference to binding means preferential binding. 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," "reduce," or "inhibition" or "inhibit" refer to a decrease or cessation of any phenotypic characteristic, or a decrease or cessation of the incidence, extent, or likelihood of that characteristic. "Reduce" or "inhibit" refers to a decrease, reduction, or prevention of an activity, function, and / or amount compared to a reference value. In some aspects, "reduce" or "inhibit" means the ability to cause an overall decrease of 20% or more. In some aspects, "reduce" or "inhibit" means the ability to cause an overall decrease of 50% or more. In some aspects, "reduce" or "inhibit" means the ability to cause an overall decrease of 75%, 85%, 90%, 95% or more. In some aspects, the amounts are inhibited or reduced over the same time period relative to a control over the same time period.

[0126] In some aspects, inhibiting a USP1 protein is inhibiting one or more activities or functions of a USP1 protein. It should be understood that one or more activities or functions of a 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 levels of inhibition 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 aspects, inhibiting a PARP protein is inhibiting one or more activities or functions of a PARP protein. It will be understood that the activity or function of one or more PARP proteins can be inhibited in vitro or in vivo. Non-limiting examples of PARP activities and functions are described herein. Exemplary levels of inhibition of the activity of one or more PARP proteins 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.

[0128] The terms "individual" or "subject" are used interchangeably herein to refer to an animal, e.g., a mammal, such as a human. In some cases, methods of treating mammals are provided, including but not limited to humans, rodents, simians, felines, canines, equines, bovines, porcines, sheep, goats, mammalian laboratory animals, mammalian farm animals, mammalian sports animals, and mammalian pets. In some instances, an "individual" or "subject" refers to an individual or subject in need of treatment for a disease or condition. In some cases, the subject being treated can be a patient, indicating that the subject has been identified as having a disease related to treatment or is at particular risk for contracting the disease.

[0129] As used herein, the terms "cancer" and "tumor" refer to or describe the physiological condition in mammals in which a population of cells is characterized by unregulated cell growth. These terms encompass both solid cancers and hematological / lymphoid cancers. Examples of cancer include, but are not limited to, cancers with defects in DNA damage repair pathways. Other examples of cancer 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 or BRCA2 wild type. The cancer may also be BRCA1 or BRCA2 mutated. The cancer may also be a PARP inhibitor-resistant or refractory cancer, or a 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 gene deletion, reduced gene expression, or the production of a gene product (e.g., a protein) with reduced or inactive activity. Loss of function mutations include, for example, missense mutations, nucleotide insertions, nucleotide deletions, and gene deletions. Loss of function mutations also include dominant negative mutations. Therefore, cancer cells in which a p53 encoding gene undergoes a loss of function mutation include cancer cells in which the p53 encoding gene contains a missense mutation as well as cancer cells that lack the p53 encoding gene. USP1 inhibitors

[0131] USP1 inhibitors have been disclosed, for example, in WO2020 / 132269 and WO2022 / 094096, each of which is herein incorporated by reference in its entirety.

[0132] In some aspects, the ubiquitin-specific processing protease 1 (USP1) inhibitors of the present disclosure include the following compounds:

[0133] or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof. In some aspects, the ubiquitin-specific processing protease 1 (USP1) inhibitor is a compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof.

[0134] 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. Publication No. 2021 / 0115049. This compound is also referred to herein as Formula I.

[0135] The chemical name of the USP1 inhibitor of Formula II 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. Publication No. 2021 / 0115049. This compound is also referred to herein as Formula II.

[0136] The chemical name of the USP1 inhibitor of Formula III 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. Publication No. 2021 / 0115049, which is incorporated herein by reference in its entirety. This compound is also referred to herein as Formula III.

[0137] In some aspects, 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 aspects, 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 aspects, the pharmaceutically acceptable acid is gentisic acid.

[0138] In some aspects, 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) is in an amorphous form. In some aspects, the amorphous form is substantially free of other polymorphic forms. In some aspects, 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%.

[0139] In another aspect, the USP1 inhibitor for the uses and methods provided herein is in solid form, which 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 aspects, the co-crystal is formed from 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 selected from the group consisting of 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, In some aspects, the pharmaceutically acceptable acid is selected from the group consisting of gentisic acid, benzoic acid, salicylic acid, and gallic acid. In some aspects, the pharmaceutically acceptable acid is selected from the group consisting of gentisic acid, benzoic acid, salicylic acid, and gallic acid. In some aspects, 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 WO2022 / 094096, which is incorporated herein by reference in its entirety.

[0140] In some aspects, 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 (gentisic acid co-crystal of Formula I) is in an amorphous form. In some aspects, the amorphous form is substantially free of other polymorphic forms. In some aspects, 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%.

[0141] In some aspects, 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) is in crystalline form.

[0142] In some aspects, 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.

[0143] In some aspects, 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.

[0144] In some aspects, 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.

[0145] In some aspects, the gentisic acid cocrystal 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 (gentisic acid cocrystal of Formula I) is in crystalline Form 2 (gentisic acid cocrystal of Formula I, Form 2 or "Form 2"). Gentisic acid cocrystal of Formula I, Form 2 is characterized by an XRPD pattern having peaks at 16.6±0.2, 18.7±0.2, and 22.5±0.2 degrees 2θ.

[0146] In some aspects, Crystalline Form 2 is an anhydrate.

[0147] In some aspects, the melting point of crystalline Form 2 is from about 184° C. to about 190° C. In some aspects, the melting point of crystalline Form 2 is from about 186° C. to about 188° C. In some aspects, the melting point of crystalline Form 2 is about 187° C.

[0148] In some aspects, Crystalline Form 2 is characterized by an XRPD pattern having peaks at 16.6 ± 0.2, 18.7 ± 0.2, and 22.5 ± 0.2 degrees 2θ when measured by Cu Ka radiation. In another aspect, Crystalline Form 2 is characterized by an XRPD pattern having peaks at 16.6 ± 0.2, 18.7 ± 0.2, 22.3 ± 0.2, and 22.5 ± 0.2 degrees 2θ when measured by Cu Ka radiation. In some aspects, Crystalline Form 2 is characterized by an XRPD pattern having peaks at 16.6 ± 0.2, 18.7 ± 0.2, 22.3 ± 0.2, 22.5 ± 0.2, and 26.0 ± 0.2 degrees 2θ when measured by Cu Ka radiation. In some aspects, crystalline Form 2 is characterized by an XRPD pattern having peaks at 16.6±0.2, 18.7±0.2, 20.8±0.2, 22.3±0.2, 22.5±0.2, and 26.0±0.2 degrees 2Θ when measured by Cu Ka radiation.

[0149] In some aspects, Crystalline Form 2 is characterized by an XRPD pattern substantially as Figure 2 In some aspects, crystalline Form 2 is characterized by three or more, four or more, five or more, or six or more XRPD peaks listed in Table 1 (a and b) below: Table 1a. Selected XRPD Peaks of Crystalline Form 2. Table 1b. XRPD peaks of crystalline form 2.

[0150] In some aspects, 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 aspects, crystalline Form 2 is characterized by an endothermic peak at about 186.0° C., as determined by DSC.

[0151] In some aspects, Crystalline Form 2 is characterized by being substantially as Figure 3 The DSC curve is shown.

[0152] In some aspects, 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 aspects, 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 aspects, Crystalline Form 2 is characterized by a loss of about 3.17 wt% between room temperature and about 170° C.

[0153] In some aspects, Crystalline Form 2 is characterized by being substantially as Figure 3 The TGA curve is shown.

[0154] In some aspects, crystalline Form 2 is characterized by at least two of the following: a) Figure 2 The XRPD pattern shown; b) as Figure 3 The DSC curve shown; or c) as Figure 3 The TGA curve is shown.

[0155] In some aspects, crystalline Form 2 has a monoclinic unit cell.

[0156] In some aspects, the unit cell of Crystalline Form 2 has about The a value, about The b value and approx. In some aspects, Form 2 has a volume of about 's unit cell.

[0157] The unit cell parameters of crystalline state 2 are shown in Table 2. Table 2. Unit cell parameters of crystalline form 2.

[0158] In some aspects, crystalline Form 2 is substantially free of other polymorphic forms. In some aspects, 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%.

[0159] In some aspects, the USP1 inhibitor comprises a mixture comprising crystalline Form 2 and a second solid state form of Formula 1. In some aspects, the second solid state form of Formula 1 is crystalline Form A. In some aspects, the USP1 inhibitor comprises a mixture comprising predominantly crystalline Form 2 as compared to other solid state forms of Formula 1. Suitable solid state forms of Formula 1 are described in WO2022 / 094096, which is incorporated herein by reference in its entirety.

[0160] In another aspect, the USP1 inhibitor is Formula II, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof. In another aspect, the USP1 inhibitor is a solid state form of Formula II, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph, or co-crystal thereof.

[0161] In some aspects, the USP1 inhibitor for the uses and methods provided herein is a solid-state 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 aspects, a 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.

[0162] Suitable solid state forms of Formula II are described in WO2022 / 094096, which is incorporated herein by reference in its entirety.

[0163] In another aspect, the USP1 inhibitor is Formula III, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof. In another aspect, the USP1 inhibitor is Formula III, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof.

[0164] In some aspects, the USP1 inhibitor for the uses and methods provided herein is a solid-state 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 aspects, a 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.

[0165] Suitable solid state forms of Formula III are described in WO2022 / 094096, which is incorporated herein by reference in its entirety.

[0166] The present disclosure covers 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 basic salts. Pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium salts, potassium salts, cesium salts, etc.; alkaline earth metal salts such as calcium salts, magnesium salts, etc.; organic amine salts such as triethylamine salts, pyridinium salts, pyridinium salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, N,N'dibenzylethylenediamine salts, etc.; inorganic acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, etc.; organic acid salts such as citrates, lactates, tartrates, maleates, fumarates, mandelates, acetates, dichloroacetates, trifluoroacetates, oxalates, formates, etc.; sulfonates such as methanesulfonates, benzenesulfonates, p-toluenesulfonates, etc.; and amino acid salts such as arginine salts, aspartates, glutamates, etc. As used herein, the term "pharmaceutically acceptable salt" refers to any salt of the USP1 inhibitors of the present disclosure, obtained, for example, by reaction with an acid or base, which salt is physiologically tolerated in the target patient (eg, a mammal, such as a human).

[0167] Methods for synthesizing USP inhibitors (including Formula I, Formula II, and Formula III) are described in U.S. Publication No. 2021 / 0115049, which is incorporated herein by reference in its entirety. Methods for preparing salts and polymorphs of USP inhibitors (including Formula I, Formula II, and Formula III) are described in PCT Publication No. WO2022 / 094096, which is incorporated herein by reference in its entirety.

[0168] In various aspects, the USP1 inhibitor reduces the level of USP1 protein and / or inhibits or reduces at least one biological activity of the USP1 protein.

[0169] In some aspects, a USP1 inhibitor specifically binds to USP1 protein. In some aspects, a USP1 inhibitor specifically binds to USP1 protein in a USP1-UAF1 complex. In some aspects, a USP1 inhibitor specifically binds to USP1 mRNA. In some aspects, a USP1 inhibitor specifically binds to USP1 protein (alone or in a USP1-UAF1 complex) or USP1 mRNA. In some aspects, a USP1 inhibitor specifically binds to UAF1 (alone or in a USP1-UAF1 complex) and inhibits or reduces the formation or activity of the USP1-UAF1 complex.

[0170] In some aspects, USP1 inhibitors reduce the formation of the USP1-UAF1 complex. In some aspects, USP1 inhibitors reduce the activity of the USP1-UAF1 complex. In some aspects, USP1 inhibitors reduce the deubiquitinase activity of USP1. In some aspects, USP1 inhibitors increase monoubiquitinated PCNA. In some aspects, USP1 inhibitors increase monoubiquitinated FANCD2. In some aspects, USP1 inhibitors increase monoubiquitinated FANCI.

[0171] In some aspects, the USP1 inhibitor does not bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or binds to deubiquitinating enzymes, 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 greater than its affinity for USP1 (i.e., the USP1 inhibitor has a K for other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) that is greater than or equal to 100-fold greater). D Comparison with USP1 K D at least 5 times, at least 10 times, at least 20 times, or at least 100 times higher).

[0172] In some aspects, the USP1 inhibitor inhibits USP1 deubiquitinase activity with an IC 50 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, e.g., as measured using the assay disclosed in U.S. Patent Application Publication No. 2017 / 0145012, or IC 50 In some aspects, the USP1 inhibitor has an IC of 50 nM to 1000 nM, for example, as measured using the assay disclosed in Liang et al., Nat Chem Biol 10:289-304 (2014). In some aspects, the USP1 inhibitor has an IC of 50 nM to 1000 nM, for example, as measured using the assay disclosed in Chen, et al., Chem Biol, 18(11):1390-1400 (2011). 50 Inhibit USP1 deubiquitinase activity. In some aspects, the USP1 inhibitor does not inhibit the activity of other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or inhibits USP1 deubiquitinase activity at an IC of 2. 50 At least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold higher than the IC 50 Inhibits the activity of other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1).

[0173] In some aspects, the USP1 inhibitors of the present disclosure bind to the USP1 protein with an affinity ranging from 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 aspects, the USP1 inhibitors of the present disclosure bind to the 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 aspects, the USP1 inhibitors of the present disclosure bind to the USP1 protein with an affinity 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 2 1 μM, about 100 nM to about 1 μM, about 300 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 aspects, the USP1 inhibitors of the present disclosure bind to the USP1 protein with an affinity 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 5 ... About 100 nM, about 50 nM to 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.

[0174] In some aspects, the USP1 inhibitors of the present disclosure bind to the 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 aspects, the USP1 inhibitors bind to the USP1 protein with an affinity of less than 1 nM.

[0175] In some aspects, the USP1 inhibitors of the present disclosure inhibit USP1 activity at an IC of 50 In some aspects, the USP1 inhibitor inhibits USP1 activity with an IC of 50 The USP1 inhibitor may be 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 aspects, the USP1 inhibitor inhibits USP1 activity with an IC of 0. 50 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 300 nM M 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.

[0176] In some aspects, the USP1 inhibitors of the present disclosure inhibit USP1 activity at an IC of 50The range is 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 1 In some aspects, the USP1 inhibitor inhibits USP1 activity at an IC of 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.008, 0.009, 0.009, 0.001, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009 ... 50 Less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some aspects, the USP1 inhibitor inhibits USP1 activity with an IC 50 Less than 1nM.

[0177] Other exemplary USP1 inhibitors are disclosed, for example, in WO 2020 / 132269 and U.S. Provisional Application No. 62 / 857,986, each of which is herein incorporated by reference in its entirety. Exemplary assay for USP1 inhibition

[0178] Any suitable assay in the art can be used to determine activity, detect results or effects, or determine efficacy. See, for example, U.S. Publication No. 2021 / 0115049, which is incorporated herein by reference in its entirety.

[0179] In some cases, determining whether a USP1 inhibitor compound inhibits USP1 deubiquitinase activity is done by measuring a mass change following cleavage of deubiquitin-bound diubiquitin. For example, ubiquitin aldehyde and ubiquitin vinyl sulfone form covalent, irreversible linkages with deubiquitinases, resulting in an observable mass change in the deubiquitinase. Similarly, cleavage of diubiquitin results in an observable mass change.

[0180] In some cases, a method for determining whether a USP1 inhibitor compound inhibits USP1 deubiquitinase activity includes 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 bond, such as ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) or ubiquitin-rhodamine 110. Such assays can also use diubiquitin containing an isopeptide bond. Exemplary diubiquitins can include a fluorophore on one ubiquitin and a quencher on the other ubiquitin, such that fluorescence increases when the diubiquitin is cleaved. Such assays can also use an enzyme-coupled system in which ubiquitin is coupled to an enzyme that becomes active and produces a fluorescent enzyme product only when released from the ubiquitin. PARP1-selective inhibitors

[0181] In various aspects, 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 the PARP1 protein.

[0182] 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 works by selectively inhibiting and trapping PARP1 at sites of DNA single-strand breaks (SSBs). This both prevents DNA repair when the DNA replication machinery collides with the PARP1-DNA non-covalent complex and leads to the generation of more harmful DNA double-strand breaks (DSBs) during DNA replication. In cases where precise DNA repair pathways are effective, such as in cells with proficient homologous recombination repair (HRR), DSBs can be accurately repaired. In contrast, in cells with defective repair pathways, such as those harboring deleterious mutations in BRCA genes, AZD5305 treatment leads to the selective accumulation of genomic instability, which ultimately selectively kills cancer cells while sparing normal cells. See PCT Application Publication No. WO2022 / 074124, which is incorporated herein by reference.

[0183] The term "AZ05305" refers to a compound with the chemical name 5-[4-[(7-ethyl-6-oxo-5H-1,5-naphthyridin-3-yl)methyl]piperazin-1-yl]-N-methyl-pyridine-2-carboxamide, the structure of which is shown below: PCT Application Publication No. WO2022 / 074124 discloses the preparation of AZD5305. In some aspects, the free base of AZD5305 is administered to a subject. In some aspects, a pharmaceutically acceptable salt of AZD5305 is administered to a subject. In some aspects, crystalline AZD5305 is administered to a subject. In some aspects, crystalline Form A AZD5305 is administered to a subject.

[0184] In one aspect, a PARP1-selective inhibitor is used with a USP1 inhibitor of the present disclosure in an anti-cancer combination therapy. In addition to the PARP1-selective inhibitor and the USP1 inhibitor, other therapies can be used before, during, or after the combination therapy. Exemplary Assays for PARP Inhibition

[0185] The present disclosure provides compounds that effectively inhibit PARP1 activity. Any suitable assay in the art can be used to determine activity, detect results or effects, or determine efficacy. See, for example, Dillon, et al., JBS., 8(3), 347-352 (2003); U.S. Patent No. 9,566,276. In some aspects, PARP1-selective inhibitors selectively inhibit and capture PARP1 in cells, for example, as determined using the PARP capture assay disclosed in Illuzzi G., et al., Clinical Cancer Research, doi: 10.1158 / 1078-0432.CCR-22-0301 (2022). In some aspects, PARP1-selective inhibitors do not effectively capture PARP1 in cells.

[0186] In some aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity at an IC of 50 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.

[0187] In some aspects, the PARP1-selective inhibitors of the present disclosure bind to the PARP1 protein with an affinity ranging from 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 aspects, the PARP1-selective inhibitors of the present disclosure bind to the 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 aspects, the PARP1-selective inhibitors of the present disclosure bind to the PARP1 protein with an affinity 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 300 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 aspects, the PARP1-selective inhibitors of the present disclosure bind to the PARP1 protein with an affinity 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 50 nM, about 50 nM to about 60 nM, about 50 nM to about 70 nM, about 50 nM to about 70 nM, about 50 nM to about 80 nM, about 50 nM to about 90 nM, about 50 nM to about 10 ... nM to about 100 nM, about 50 nM to 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 aspects, the PARP1-selective inhibitors of the present disclosure bind to the 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 aspects, the PARP1-selective inhibitors of the present disclosure bind to the PARP1 protein with an affinity of less than 1 nM.

[0188] In some aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity at an IC of 50 In some aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity with an IC of 50 The PARP1 activity is 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 aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity with an IC of 0. 50 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 300 nM M 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 aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity with an IC of 0. 50The range is 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 1 In some aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity with an IC of 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.008, 0.009, 0.009, 0.001, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009 ... 50 Less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some aspects, the PARP1-selective inhibitors of the present disclosure inhibit PARP1 activity with an IC 50 Less than 1nM.

[0189] In some aspects, the PARP1-selective inhibitor is at least 500-fold selective for PARP1 over PARP2. How to use

[0190] Since the combination of the present disclosure is an inhibitor of USP1 protein and PARP1 protein, the present disclosure provides a method for inhibiting USP1 protein and / or PARP1 protein, which comprises contacting USP1 and / or PARP1 protein or a composition comprising USP1 and / or PARP1 protein with one or more combinations of the present disclosure.

[0191] Because the combinations of the present disclosure are inhibitors of USP1 and PARP1 proteins, a variety of diseases, conditions, or disorders mediated by USP1 and / or PARP1 proteins can be treated using these compounds. Accordingly, the present disclosure generally relates 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 developing such a condition, the method comprising administering to the animal an effective amount of one or more combinations of the present disclosure.

[0192] The present disclosure also relates to a method of inhibiting USP1 and / or PARP1 protein in an animal in need thereof, the method comprising administering to the animal a therapeutically effective amount of a combination of the present disclosure.

[0193] In some aspects, the disclosed combinations can be used to inhibit the activity of USP1 and / or PARP1 proteins. For example, in some aspects, a method of inhibiting USP1 and / or PARP1 proteins comprises contacting USP1 and / or PARP1 proteins with the disclosed combinations. The contacting can occur in vitro or in vivo.

[0194] In some aspects, the disclosed combinations can be used to treat diseases mediated by USP1 and / or PARP1 proteins. A disease mediated by USP1 and / or PARP1 proteins is any pathological condition in which USP1 and / or PARP1 proteins are known to play a role. In some aspects, the USP1 and / or PARP1 mediated disease is a proliferative disease, such as cancer. In some aspects, the disclosed combinations can delay, reduce, or prevent rebound (rapid regrowth) of a tumor.

[0195] In some aspects, the disclosed combinations are less toxic than the PARP1-selective inhibitor alone (eg, due to administration of less PARP1-selective inhibitor).

[0196] Provided herein are various methods of treating diseases and disorders using the disclosed combinations. Exemplary diseases and disorders that can be treated with the disclosed combinations include, but are not limited to, cancer.

[0197] In some aspects, methods of treating cancer using the disclosed combinations are provided. Such methods comprise administering a therapeutically effective amount of the disclosed combination to a subject suffering from cancer.

[0198] In some aspects, the cancer to be treated with the combination of the present disclosure is selected from a blood cancer, a lymphoma, and a cancer with a defect in a DNA damage repair pathway. In some aspects, the cancer to be treated with the combination of the present disclosure is a cancer comprising cancer cells having a mutation in the gene encoding p53. In some aspects, the cancer to be treated with the combination of the present disclosure is a cancer comprising cancer cells having a loss-of-function mutation in the gene encoding p53. In some aspects, the cancer to be treated with the combination of the present disclosure is a cancer comprising cancer cells having a mutation in the gene encoding BRCA1. In some aspects, the cancer to be treated with the combination of the present disclosure is a cancer comprising cancer cells having a mutation in the gene encoding BRCA2. In some aspects, the cancer to be treated with the combination of the present disclosure is a cancer comprising cancer cells having a loss-of-function mutation in the gene encoding ATM.

[0199] In some aspects, the cancer to be treated with the combination of the present disclosure is an advanced solid tumor. Advanced solid tumors are solid tumors that are unresectable and / or metastatic. In some aspects, the cancer to be treated with the combination of the present disclosure is a solid tumor that is a resectable tumor, whether benign or metastatic, wherein the combination is administered before and / or after resection surgery.

[0200] In some aspects, the cancer 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 aspects, the cancer is uterine cancer. In some aspects, the cancer is peritoneal cancer. In some aspects, the cancer is endometrial cancer. In some aspects, the cancer is ovarian cancer or breast cancer. In some aspects, the cancer is ovarian cancer. In some aspects, the cancer is breast cancer. In some aspects, the cancer is triple-negative breast cancer. In some aspects, the cancer is ovarian cancer. In some aspects, the ovarian cancer is a BRCA1 mutation cancer, a BRCA2 mutation cancer, or a p53 mutation cancer. In some aspects, the ovarian cancer is a BRCA1 mutation cancer and a p53 mutation cancer. In some aspects, the ovarian cancer is a BRCA1 and BRCA2 mutation cancer. In some aspects, the ovarian cancer is a BRCA2 mutation cancer.

[0201] In some aspects, the cancer to 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 carcinoma 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 aspects, the cancer to be treated with the combination of the present disclosure is selected from the group consisting of uterine cancer, peritoneal cancer and endometrial cancer.

[0202] Provided herein are various methods of treating cancer using the disclosed combinations.In some aspects, a therapeutically effective amount of a disclosed combination is administered to a subject having cancer.

[0203] In some aspects, such methods comprise administering a therapeutically effective amount of a combination of the present disclosure to a subject having triple-negative breast cancer.

[0204] In some aspects, the combination of the present disclosure is used to treat cancer, wherein the cancer is a homologous recombination defective cancer. In some aspects, the combination of the present disclosure is used to treat cancer, wherein the cancer includes cancer cells with p53 encoding gene mutations. In some aspects, the combination of the present disclosure is used to treat cancer, wherein the cancer includes cancer cells with loss of function mutations in p53 encoding genes. In some aspects, the combination of the present disclosure is used to treat cancers that do not have defects in the homologous recombination pathway.

[0205] In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is a BRCA1 mutation cancer. In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is a BRCA2 mutation cancer. In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is a BRCA1 mutation cancer and a BRCA2 mutation cancer. In some aspects, the cancer is not a BRCA1 mutation cancer or a BRCA2 mutation cancer. In some aspects, the cancer is a BRCA1 deficiency cancer. In some aspects, the cancer is a BRCA2 deficiency cancer. In some aspects, the cancer is a BRCA1 deficiency cancer and a BRCA2 mutation cancer.

[0206] In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is an ATM mutant cancer. In some aspects, the cancer is not an ATM mutant cancer. In some aspects, the cancer is an ATM deficient cancer.

[0207] In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is a PARP inhibitor-resistant or refractory cancer. In some aspects, the combinations of the present disclosure are used to treat cancer, wherein the cancer is a PARP inhibitor-resistant or refractory BRCA1-deficient cancer. In some aspects, the cancer is an olaparib-resistant or refractory cancer. In some aspects, the cancer is an AZD-5305-resistant or refractory cancer.

[0208] In some aspects, the cancer is a BRCA1 and / or BRCA2 mutant cancer, wherein the cancer comprises cells with elevated RAD18 levels, e.g., wherein elevated RAD18 levels are at least as high as RAD18 protein and / or mRNA levels in ES2 cells (ES2 cells are publicly available, e.g., available from American Type Culture Collection (ATCC; CRL-1978)), or wherein elevated RAD18 levels are greater than RAD18 protein and / or mRNA levels in HEP3B217 cells (HEP3B217 cells are publicly available, e.g., available from ATCC (HB-8064)). In some aspects, triple-negative breast cancer is a BRCA1 and / or BRCA2 mutant cancer.

[0209] In some cases, methods described herein include detecting RAD51 (e.g., RAD51 protein, RAD51 protein foci and / or RAD51 mRNA) levels in cancer cells (e.g., using samples obtained from cancer). RAD51 protein levels can be detected using, for example, immunofluorescence, Western blotting, fluorescence activated cell sorting (FACS) and / or immunohistochemistry. For example, RAD51 mRNA levels can be detected using quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR).

[0210] In some aspects, the combinations of the present disclosure are used to treat cancers previously treated with platinum therapy. In some aspects, the combinations of the present disclosure are used to treat platinum-resistant cancers. In some aspects, the combinations of the present disclosure are used to treat platinum-refractory cancers. In some aspects, the platinum therapy is selected from the group consisting of cisplatin, carboplatin, oxaliplatin, and combinations thereof.

[0211] In some cases, cancer is a solid cancer. In some cases, cancer is a blood cancer / lymphoma. In some cases, cancer is a cancer with a defective DNA damage repair pathway. In some cases, cancer is a homologous recombination defective cancer. In some cases, cancer includes cancer cells with a p53 encoding gene mutation. In some cases, cancer includes cancer cells with a p53 encoding gene loss of function mutation. In some cases, 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, cancer is ovarian cancer or breast cancer (including triple-negative breast cancer). In some cases, cancer is ovarian cancer. In some cases, cancer is breast cancer (including triple-negative breast cancer). In some cases, cancer is uterine cancer. In some cases, cancer is peritoneal cancer. In some cases, cancer is endometrial cancer.

[0212] In some aspects, the combinations of the present disclosure are used in combination with one or more additional therapeutic agents to treat cancer.

[0213] In some aspects, provided herein are combinations of the present disclosure for use as a medicament or for the preparation of a medicament, e.g., for the treatment of cancer. In some aspects, provided herein are combinations of the present disclosure for use in a method of treating cancer. Combination pharmaceutical composition

[0214] The combination of the present disclosure can be administered to a mammal in the form of the raw chemical without any other components, or the combination of the present disclosure can also be administered to a mammal as part of a pharmaceutical composition comprising the compound in combination with a suitable pharmaceutically acceptable carrier (see, for example, 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 a carrier can be selected from pharmaceutically acceptable excipients and adjuvants. The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable solvent" encompasses any standard pharmaceutical carrier, solvent, surfactant or vehicle. 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 may be present in the same pharmaceutical composition; alternatively, the USP1 inhibitor and the PARP1-selective inhibitor may be contained in separate pharmaceutical compositions that can be administered simultaneously or sequentially.

[0215] The combined pharmaceutical compositions of the present disclosure can be prepared as liquid suspensions or solutions using liquids such as oils, water, alcohol, and combinations thereof.

[0216] As provided herein, USP1 inhibitors (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 included in any pharmaceutical composition, including all compositions comprising such inhibitors in combination with one or more pharmaceutically acceptable carriers. In some aspects, the USP1 inhibitor is present in the composition in an amount effective to achieve its intended therapeutic purpose.

[0217] In some aspects, the present 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 aspects, the pharmaceutical composition comprises crystalline State 2 of the gentisic acid cocrystal of Formula I.

[0218] The combined pharmaceutical compositions for in vivo administration may be sterile. This can be readily accomplished by filtration, for example, through sterile filtration membranes.

[0219] Combination pharmaceutical compositions within the scope of the present disclosure include all compositions of the USP1 inhibitors and PARP1-selective inhibitors of the present disclosure in combination with one or more pharmaceutically acceptable carriers. In one aspect, the USP1 inhibitors and PARP1-selective inhibitors of the present disclosure are present in the composition in amounts effective to achieve their intended therapeutic purpose.

[0220] The combined pharmaceutical compositions of the present disclosure can be administered to any patient who may experience the beneficial effects of the combinations of the present disclosure. Foremost among such patients are mammals, such as humans and pets, although the present disclosure is not intended to be limited thereto. In one aspect, the patient is a human. In another aspect, the combined pharmaceutical compositions of the present disclosure can be administered to a patient with a PARP inhibitor-resistant or refractory cancer. In another aspect, the combined pharmaceutical compositions of the present disclosure can be administered to a patient with a PARP inhibitor-resistant or refractory BRCA1-deficient cancer.

[0221] In another aspect, the present disclosure provides a kit comprising a combination of the present disclosure, the combination being packaged in a manner that facilitates its use in practicing the methods of the present disclosure. In one aspect, the kit comprises a USP1 inhibitor and a PARP1-selective inhibitor of the present disclosure packaged in a container (e.g., a sealed bottle or vessel), wherein a label is affixed to the container or contained in the kit, the label describing the use of the compound to practice the methods of the present disclosure. In one aspect, the combined composition is packaged in unit dosage form. The kit may also include a device suitable for administering the combined composition according to the intended route of administration. In some aspects, the present disclosure provides a kit comprising a USP1 inhibitor and a PARP1-selective inhibitor of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, and instructions for administering the compound, or a pharmaceutically acceptable salt or solvate thereof, to a patient suffering from cancer.

[0222] In some aspects, the present disclosure provides a combined pharmaceutical 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.

[0223] In some aspects, the present disclosure provides a combined pharmaceutical 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 a USP1 gene and / or a PARP1 gene.

[0224] In some aspects, the present disclosure provides a combined pharmaceutical 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 used to treat cancer.

[0225] In some aspects, the present disclosure provides a combined pharmaceutical 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 used in the manufacture of a medicament for treating cancer. Example Example 1: Antitumor activity of the cocrystal of Formula I in combination with the selective PARP1 inhibitor AZD5305 in a patient-derived breast xenograft model in nude mice

[0226] The USP1 inhibitor used in the examples is the co-crystal described in paragraphs

[140] and

[0143] -

[0155] above. The co-crystal can be prepared according to Example 6 of the published PCT WO2022 / 094096. The co-crystal of Formula I 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.

[0227] like Figures 1A to 1C As shown, the antitumor activity of the USP1 inhibitor of Formula I cocrystal in combination with AZD5305 in nude mice was evaluated using a patient-derived mammary gland xenograft model resistant to olaparib. Female athymic nude mice aged 6-9 weeks from Envigo were anesthetized and a 20 mm incision was made in the flank. 3 Tumor fragments were placed subcutaneously. When the tumors were established and the tumor volume ranged from 63 to 196 mm 3Mice were randomly divided into 10 groups 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). Depending on the treatment group, the compounds were administered once daily by oral gavage for at least 42 days. Body weight and tumor volume were measured twice weekly. Tumor volume was calculated as the mean and standard error of the mean for each treatment group.

[0228] Figure 1A The data in show that the combination treatment group exhibited enhanced anti-tumor activity in a patient-derived subcutaneous mouse model compared to equivalent doses of single agents of Formula I or AZD5305.

[0229] To evaluate the activity of the combination in mice with tumors resistant to PARP inhibitors, five mice that showed resistance to AZD5305 monotherapy were subsequently dosed with a combination of Formula I (100 mg / kg) and AZD5305 (10 mg / kg) on ​​treatment day 49. The addition of Formula I resulted in a reduction in tumor volume compared to the remaining animals treated with AZD5305 monotherapy, as shown in Figure 1B.

[0230] 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 change in body weight as a percentage relative to the body weight on the day treatment started (Day 0), as Figure 1C shown. Example 2: In vitro efficacy

[0231] In vitro experiments were performed using the colony forming unit (CFU) assay. UWB1.289 parental or UWB1.289 BRCA1 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 visible in the 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 dye in each well was quantified by extracting the crystal violet into 10% acetic acid and measuring the absorbance at 565 nm. The CFU results are shown in Figure 2. Figure 4 and Figure 5 The results indicate that a USP1 inhibitor and a PARP1-selective inhibitor are an effective combination for UWB1.289 and UWB1.289BRCA1-overexpressing cells.

[0232] Having now fully described the invention, one of ordinary skill in the art will appreciate that the invention can be practiced within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any aspect thereof.

[0233] Other aspects of the 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 exemplary only, with the true scope and spirit of the invention being indicated by the following claims.

[0234] All patents and publications cited herein are incorporated by reference in their entirety.

Claims

1. A method of treating cancer in a subject, the method comprising administering to the subject (i) an ubiquitin-specific processing protease 1 (USP1) inhibitor and (ii) a poly ADP-ribose polymerase 1 (PARPl)-selective inhibitor, or a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof, wherein the USP1 inhibitor is a compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof.

2. A method for delaying, reducing or preventing tumor rebound in a subject, the method 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, wherein the USP1 inhibitor is a compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids, polymorphs or co-crystals thereof.

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. 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. 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 a pharmaceutically acceptable acid and the USP1 inhibitor.

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, optionally wherein the pharmaceutically acceptable acid is a gentisic acid, benzoic acid or salicylic acid co-crystal.

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

10. The method of any one of claims 1 to 9, wherein the USP1 inhibitor, or the pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof, and the PARP1-selective inhibitor, or the pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof are administered in therapeutically effective amounts sufficient to produce one or more therapeutic effects selected from the group consisting of: (i) reducing tumor size, (ii) increasing tumor regression rate, and (iii) reducing or inhibiting tumor growth.

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

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

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

14. The method of any one of claims 1 to 13, wherein the USP1 inhibitor and / or the PARP1-selective inhibitor is administered at a dose that is ineffective as a single agent.

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

16. A combined composition comprising (i) an 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 compound selected from the group consisting of: (a) Formula I: (b) Formula II: (c) Formula III: and pharmaceutically acceptable salts, hydrates, solvates, amorphous solids or polymorphs thereof.

17. The composition of claim 16, wherein the PARP1-selective inhibitor is AZD5305 and a pharmaceutically acceptable salt, hydrate, solvate, amorphous solid, polymorph or co-crystal thereof.

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

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

20. The composition of claim 16 or claim 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 a USP1 inhibitor, optionally 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, optionally wherein the co-crystal is a gentisic acid, benzoic acid or salicylic acid co-crystal.

22. The composition of 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 in the preparation of a medicament for treating cancer.

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

26. The pharmaceutical composition of claim 25, for use in treating 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 suffering from cancer.

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

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

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, endometrial cancer and breast cancer.

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

33. The method, use, pharmaceutical composition or kit of any one of claims 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 26-34, wherein the cancer is a DNA damage repair pathway defective cancer.

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 of any one of claims 1 to 15, 24 or 26 to 38, wherein the cancer is a BRCA1 mutation cancer and a BRCA2 mutation 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. 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 a gene encoding ATM.

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 been previously treated with platinum therapy, optionally wherein the cancer is platinum-resistant or platinum-refractory.

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 disease mediated by USP1 protein and / or a disease mediated by PARP1 protein, the method comprising administering to a subject in need thereof an effective amount of the composition of any one of claims 16 to 23, or the pharmaceutical composition of claims 25, 26, or 28 to 44, to treat the disease mediated by USP1 protein and / or the disease mediated by PARP1 protein.

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

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

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

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