Methods of treating cancers having FBXW7 mutations

Abstract

A method for treating a cancer in a subject, the method comprising administering a therapeutically effective amount of a membrane- associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitor, wherein the cancer comprises a truncating biallelic loss- of-function mutation in FBXW7 is disclosed. In a preferred embodiment, the Myt1 inhibitor is a compound of Formula (I). TheMyt1 inhibitors may bee used in combination with other therapeutic agents. Such agents include Wee 1l inhibitor, Fen 1 inhibitor, Top 1 inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Aurkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod 1 inhibitor, Sod2 inhibitor, Bub 1 inhibitor, Cdc7 inhibitor, Sae 1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Out inhibitor, Hdac3 inhibitor, Chek1 inhibitor, Aurka inhibitor, Men 1 inhibitor, Dot 1 I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinum-based DNA damaging agent, or a combination thereof.

Description

[0001] ATTORNEY DOCKET NO.: D0963.70010WO00

[0002] METHODS OF TREATING CANCERS HAVING FBXW7 MUTATIONS

[0003] FIELD OF THE INVENTION

[0004] The invention relates to therapies using inhibitors of membrane associated tyrosine and threonine specific Cdc2 inhibitory kinase (Myt1) (gene name PKMYTT) to treat cancers having a truncating biallelic loss-of-function mutation ora loss-of-function missense mutation described herein.

[0005] BACKGROUND

[0006] F-box / WD Repeat-Containing Protein 7 (FBXW7) is one of the four subunits of the ubiquitin protein ligase complex (the “Skp, Cullin, F-box containing complex” or the “SCF” complex) and, through its F-box domain, imparts the SCF complex with protein-protein interactions. FBXW7 is particularly noted for binding to, and therefore promoting the ubiquitylation of, Cyclin E1 (CCNE1; encoded for by the CCNE1 gene). Some mutations to FBXW7 can lower the binding efficiency of FBXW7 to CCNE1, resulting in CCNE1 overexpression in cells. CCNE1 over-expression and / or FBXW7 loss-of-function is thought to drive tumorigenesis by inducing genome instability (e.g., increased origin firing, defective nucleotide pools, transcription-replication conflicts, and / or fork instability). The present disclosure highlights the discovery that specific mutations in FBXW7, particularly truncating biallelic loss-of-function mutations or loss-of-function missense mutations (e.g., missense mutations encoding an amino acid substitution selected from R465, R479, and / or R505), impart great sensitivity to cancer treatment with membrane-associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitors. This discovery addresses a need in the field for new anticancer therapeutic methods and, in particular, those targeting patient populations particularly susceptible to an anti-cancer therapy.

[0007] SUMMARY OF THE INVENTION

[0008] In general, the present disclosure relates to the discovery of the susceptibility of cells (e.g., cancer cells, e.g., tumors) harboring (i) truncating biallelic loss-of-function mutations (e.g., cells harboring a truncating mutation in a first allele and a missense mutation in a second allele of the same gene, or cells harboring a truncating mutation in a first allele and a truncating mutation in a second allele of the same gene) or (ii) a loss-of-function missense mutations (e.g., missense mutations encoding an amino acid substitution selected from R465, R479, and / or R505) in genes which encode for F-box proteins (e.g., FBXW7, which encodes F-box / WD repeat containing protein (FBXW7)) to treatment with a membrane-associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitors. For example, the present disclosure highlights that the treatment of cancer cells having specific mutations in FBXW7 can increase the cells sensitivity to treatment with Myt1 inhibitors.

[0009] In a first aspect, the disclosure provides a method for treating a cancer in a subject, the method including administering to the subject in need thereof a therapeutically effective amount of a membrane-associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitor, wherein the cancer includes a truncating biallelic loss-of-function mutation in FBXW7. ATTORNEY DOCKET NO.: D0963.70010WO00

[0010] In a second aspect, the disclosure provides a method for treating a cancer in a subject, the method including administering to the subject in need thereof a therapeutically effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having a truncating biallelic loss-of-function mutation in FBXW7.

[0011] In a third aspect, the disclosure provides method of inducing cell death in a cancer cell having a truncating biallelic loss-of-function mutation in FBXW7, the method including contacting the cell with an effective amount of a Myt1 inhibitor.

[0012] In a fourth aspect, the disclosure provides a method of inducing cell death in a cancer cell, the method including administering to a subject in need thereof a thereof, an effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having a truncating biallelic loss-of-function mutation in FBXW7.

[0013] In some embodiments of any of the foregoing aspects, the truncating biallelic loss-of-function mutation includes one or two truncating mutations selected from the group consisting of a nonsense mutation, a nonstop mutation, a frameshift mutation, and a splice site mutation. In some embodiments, the truncating mutation produces an FBXI / l / 7transcriptthat is 50-1800 nucleotides in length (e.g., 50-1700 nucleotides, 50-1600 nucleotides, 50-1500 nucleotides, 50-1400 nucleotides, 50-1300 nucleotides, 50-1200 nucleotides, 100-1850 nucleotides, 150-1800 nucleotides, 200-1750 nucleotides, 250-1700 nucleotides, 300-1650 nucleotides, 350-1600 nucleotides, 400-1550 nucleotides, 450-1500 nucleotides, 500-1450 nucleotides, 550-1400 nucleotides, 600-1350 nucleotides, 650-1300 nucleotides, or 700-1250 nucleotides in length. In some embodiments, the truncating mutation produces an FBXW7 polypeptide that is 50-600 amino acids, 50-550 amino acids, 50-500 amino acids, 50-450 amino acids, 50-400 amino acids, 50-350 amino acids, 50-300 amino acids, 50-250 amino acids, 50-200 amino acids, 50-150 amino acids, 50-100 amino acids, 100-600 amino acids, 150-550 amino acids, 200-500 amino acids, 250-450 amino acids, or 300-400 amino acids in length.

[0014] In some embodiments of any of the foregoing aspects, the truncating biallelic loss-of-function mutation includes one truncating mutation (e.g., in a first allele of FBXW7). In other embodiments, the truncating biallelic loss-of-function mutation includes two truncating mutations (e.g., in both alleles of FBXW7). In yet other embodiments, the truncating biallelic loss-of-function mutation includes (i) one truncating mutation (e.g., in a first allele of FBXW7) and at least one other mutation (e.g., a missense mutation, splice site, or another truncating mutation in a second allele of FBXW7) or (ii) one truncating mutation (e.g., in a first allele of FBXW7) and a partial or complete deletion of a second allele of FBXW7 (e.g., a loss-of-heterozygosity in FBXW7). In some embodiments, the at least one other mutation is a missense mutation (e.g., a hotspot mutation in a second allele of FBXW7). In some embodiments, the at least one other mutation (e.g., missense mutation) encodes for an amino acid substitution in FBXW7 selected from the group consisting of an arginine (R) at amino acid position 465 (R465), an R at amino acid position 479 (R479), and an R at amino acid position 505 (R505).

[0015] In a fifth aspect, the disclosure provides a method for treating a cancer in a subject, the method including administering to the subject in need thereof a therapeutically effective amount of a ATTORNEY DOCKET NO.: D0963.70010WO00

[0016] Myt1 inhibitor, wherein the cancer includes at least one missense mutation in FBXW7. In some embodiments, the at least one missense mutation in FBXW7 encodes for an amino acid substitution in FBXW7 selected from the group consisting of R465, R479, and R505. In some embodiments, the amino acid substitution is R465. In some embodiments, the amino acid substitution is R479. In some embodiments, the amino acid substitution is R505. In some embodiments, the amino acid substitution results in a loss-of-function of FBXW7.

[0017] In a sixth aspect, the disclosure provides a method for treating a cancer in a subject, the method including administering to the subject in need thereof a therapeutically effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having at least one missense mutation in FBXW7. In some embodiments, the at least one missense mutation in FBXW7 encodes for an amino acid substitution in FBXW7 selected from the group consisting of R465, R479, and R505. In some embodiments, the amino acid substitution is R465. In some embodiments, the amino acid substitution is R479. In In some embodiments, the amino acid substitution is R505. In some embodiments, the amino acid substitution results in a loss-of-function of FBXW7.

[0018] In a seventh aspect, the disclosure provides a method of inducing cell death in a cancer cell having at least one missense mutation in FBXW7, the method including contacting the cell with an effective amount of a Myt1 inhibitor. In some embodiments, the at least one missense mutation in FBXW7 encodes for an amino acid substitution in FBXW7 selected from the group consisting of R465, R479, and R505. In some embodiments, the amino acid substitution is R465. In some embodiments, the amino acid substitution is R479. In In some embodiments, the amino acid substitution is R505. In some embodiments, the amino acid substitution results in a loss-of-function of FBXW7.

[0019] In an eighth aspect, the disclosure provides a method of inducing cell death in a cancer cell, the method including administering to a subject in need thereof a thereof an effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having at least one missense mutation in FBXW7. In some embodiments, the at least one missense mutation in FBXW7 encodes for an amino acid substitution in FBXW7 selected from the group consisting of R465, R479, and R505. In some embodiments, the amino acid substitution is R465. In some embodiments, the amino acid substitution is R479. In In some embodiments, the amino acid substitution is R505. In some embodiments, the amino acid substitution results in a loss-of-function of FBXW7.

[0020] In some embodiments of any of the foregoing aspects, the cancer is colorectal cancer. In some embodiments of any of the foregoing aspects, the cancer is bladder cancer. In some embodiments of any of the foregoing aspects, the cancer is cervical cancer. In some embodiments of any of the foregoing aspects, the cancer is upper gastrointestinal cancer. In some embodiments, the cancer is further characterized by CCNE1 overexpression.

[0021] In some embodiments of any of the foregoing aspects, the method further includes identifying the cancer as having CCNE1 overexpression.

[0022] In some embodiments of the first or third aspect, the method further includes identifying the cancer or the cancer cell as having the truncating biallelic loss-of-function mutation in FBXW7. In some embodiments of the fifth or seventh aspect, the method further includes identifying the cancer ATTORNEY DOCKET NO.: D0963.70010WO00

[0023] or cancer cell as having the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7. In some embodiments, identifying the cancer or cancer cell as having (i) the truncating biallelic loss-of-function mutation in FBXW7, or (ii) the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7, includes targeted panel sequencing, whole genome sequencing (WGS) or Whole Exome Sequencing (WES). In some embodiments, identifying the cancer or cancer cell as having (i) the truncating biallelic loss-of-function mutation in FBXW7, or (ii) the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7, includes:

[0024] from read counts fora plurality of single nucleotide variants (SNVs) including homozygous and heterozygous SNVs obtained from sequencing a sample including the cancer cell and from reference read counts, determining an integer total copy number of a locus segment within an FBXW7 gene region in a cancer cell from the subject or in the cancer cell and / or two integer allele-specific copy numbers of the locus segment,

[0025] wherein the cancer is identified as having (i) the truncating biallelic loss-of-function mutation in FBXW7, or (ii) the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7,

[0026] if at least one of the integer total copy number and the integer allele-specific copy numbers is 0, provided that the remaining FBXW7 allele, if present, includes an inactivating mutation, or

[0027] if none of the integer allele-specific copy numbers is 0 and FBXW7 alleles are present, each of the FBXW7 alleles independently having an inactivating mutation.

[0028] In some embodiments, the determining step includes:

[0029] from the read counts and the reference read counts, determining total copy number log-ratios, allelic copy number log-odds ratios, and target coverage values for the SNVs;

[0030] segmenting the total copy number log-ratios and the allelic copy number log-odds ratios; estimating sample purity and sample ploidy for the cancer cell from the total copy number logratios and the target coverage values; and

[0031] from the target coverage values, the sample purity, the sample ploidy, the total copy number log-ratios, and the allelic copy number log-odds ratios, generating an integer total copy number of a segment including a plurality of SNVs within an FBXW7 gene region in the cancer cell and two integer allele-specific copy numbers of the segment.

[0032] In some embodiments, the method further includes adjusting the ratios for location shift. In some embodiments, the plurality of SNVs includes consistently covered SNVs. In some embodiments, each of the consistently covered SNVs has the mean coverage of at least 200x reads across panel of normal samples. In some embodiments, the plurality of SNVs includes frequent SNVs, the frequent SNVs having an allele frequency of 33% to 66% in humans. In some embodiments, the plurality of SNVs includes SNVs proximal to the frequent SNVs. In some embodiments, the plurality of SNVs includes SNVs, each of the SNVs having a 5’-flanking sequence of at least 20 contiguous nucleobases including 25-75% GC content, wherein the 5’-flanking sequence is unique and does not include other SNVs. In some embodiments, the plurality of SNVs ATTORNEY DOCKET NO.: D0963.70010WO00

[0033] includes at least 20 heterozygous SNVs. In some embodiments, the reference read counts are from a panel of normal samples.

[0034] In some embodiments of any of the foregoing aspects, the FBXW7 gene region is an F-box region. In some embodiments, the FBXW7 gene region is a WD40 repeat.

[0035] In some embodiments of any of the foregoing aspects, the truncating biallelic loss-of-function mutation in FBXW7 or the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7 includes at least one somatic FBXW7 loss-of-function mutation. In some embodiments of any of the foregoing aspects, the truncating biallelic loss-of-function mutation in FBXW7 or the missense mutation encoding the substitution mutation (e.g., R465, R479, and / or R505) in FBXW7 includes at least one germline FBXW7 loss-of-function mutation.

[0036] In some embodiments of any of the foregoing aspects, the method further includes administering a therapeutically effective amount of a Wee1 inhibitor, Fen1 inhibitor, Topi inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Aurkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod1 inhibitor, Sod2 inhibitor, Bub1 inhibitor, Cdc7 inhibitor, Sae1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Dut inhibitor, Hdac3 inhibitor, Chekl inhibitor, Aurka inhibitor, Men1 inhibitor, Doti I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinumbased DNA damaging agent, or a combination thereof.

[0037] In some embodiments, the method includes administering the Wee1 inhibitor. In some embodiments, the Wee1 inhibitor is one of:

[0038]

[0039] In some embodiments, the method includes administering the Fen1 inhibitor. In some embodiments, the Fen1 inhibitor is C8 (PMID: 32719125), SC13, Fen1-IN-3, or a pharmaceutically acceptable salt thereof. ATTORNEY DOCKET NO.: D0963.70010WO00

[0040] In some embodiments, the method includes administering the Topi inhibitor. In some embodiments, the Topi inhibitor is irinotecan, topotecan, camptothecin, lamellarin D, or a pharmaceutically acceptable salt thereof.

[0041] In some embodiments, the method includes administering the Rrm1 inhibitor. In some embodiments, the method includes administering the Rrm2 inhibitor. In some embodiments, the Rrm2 inhibitor is motexafin gadolinium, hydroxyurea, fludarabine, cladribine, tezacitabine, triapine, or a pharmaceutically acceptable salt thereof.

[0042] In some embodiments, the method includes administering the Aurkb inhibitor. In some embodiments, the Aurkb inhibitor is MK0547, AZD1152, PHA739358, AT9283, AMG900, SNS-314, TAK-901, 2CYC116, GSK1070916, PF03814735, or a pharmaceutically acceptable salt thereof.

[0043] In some embodiments, the method includes administering the Top2A inhibitor. In some embodiments, the Top2A inhibitor is etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, or a pharmaceutically acceptable salt thereof.

[0044] In some embodiments, the method includes administering the ATR inhibitor. In some embodiments, the ATR inhibitor is a compound of formula (IV):

[0045] ^N^ R1

[0046]

[0047] (IV),

[0048] or a pharmaceutically acceptable salt thereof,

[0049] wherein

[0050] - is a double bond, and each Y is independently N or OR4; or - is a single bond, and each Y is independently NRY, carbonyl, or C(RY)2; wherein each RYis independently H or optionally substituted C1-6 alkyl;

[0051] R1is optionally substituted C1-6 alkyl or H;

[0052] R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2,-SO2R5A, or-Q-R5B;

[0053] R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl; each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl;

[0054] each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, or–SO2R5A; or ATTORNEY DOCKET NO.: D0963.70010WO00

[0055] both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0056] each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted C6-10 aryl;

[0057] R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or optionally substituted alkoxy;

[0058] each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0059] Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted C6-10 arylene; and

[0060] X is hydrogen or halogen.

[0061] In some embodiments, the ATR inhibitor is a compound of formula (V):

[0062] N^R1

[0063]

[0064] (V),

[0065] or a pharmaceutically acceptable salt thereof,

[0066] wherein

[0067] each Y is independently N or CR4;

[0068] R1is optionally substituted C1-6 alkyl or H;

[0069] R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2,-SO2R5A, or-Q-R5B;

[0070] R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl; each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl;

[0071] each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, or–SO2R5A; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0072] each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted C6-10 aryl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0073] R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, –SO2R5A, or optionally substituted alkoxy;

[0074] each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted Ce-io aryl C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0075] Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted Ce-io arylene; and

[0076] X is hydrogen or halogen.

[0077] In some embodiments, the ATR inhibitor is selected from the group consisting of compounds A43, A57, A62, A87, A93, A94, A95, A99, A100, A106, A107, A108, A109, A111, A112, A113, A114, A115, A116, A118, A119, A120, A121 (camonsertib), A122, A123, A135, A147, A148, and pharmaceutically acceptable salts thereof. In some embodiments, the ATR inhibitor is compound A43 or a pharmaceutically acceptable salt thereof. In some embodiments, the ATR inhibitor is camonsertib or a pharmaceutically acceptable salt thereof. In some embodiments, the ATR inhibitor is compound A122 or a pharmaceutically acceptable salt thereof. In some embodiments, the ATR inhibitor is

[0078]

[0079] elimusertib (BAY1895344) ceralasertib (AZD6738) berzosertib (VE-822)

[0080]

[0081] Gartisertib (VX-803, M4344) tuvusertib (M1774) ATTORNEY DOCKET NO.: D0963.70010WO00

[0082]

[0083] SKLB-197

[0084] or a pharmaceutically acceptable salt thereof.

[0085] In some embodiments, the method includes administering the Ttk inhibitor. In some embodiments, the Ttk inhibitor is BAY1217389 or a pharmaceutically acceptable salt thereof.

[0086] In some embodiments, the method includes administering the Sod1 inhibitor. In some embodiments, the Sod1 inhibitor is LCS1, ATN-224, Pyrimethamine, a compound of the following structure

[0087]

[0088] or a pharmaceutically acceptable salt thereof.

[0089] In some embodiments, the method includes administering the Sod2 inhibitor. In some embodiments, the Sod2 inhibitor is LCS1, ATN-224, pyrimethamine, or a pharmaceutically acceptable salt thereof.

[0090] In some embodiments, the method includes administering the Bub1 inhibitor. In some embodiments, the Bub1 inhibitor is BAY-320, BAY-419, BAY1816032, or a pharmaceutically acceptable salt thereof.

[0091] In some embodiments, the method includes administering the Cdc7 inhibitor. In some embodiments, the Cdc7 inhibitor is SRA141, TAK931, or a pharmaceutically acceptable salt thereof.

[0092] In some embodiments, In some embodiments, the method includes administering the Sae1 inhibitor. In some embodiments, the Sae1 inhibitor is ML792 or a pharmaceutically acceptable salt thereof.

[0093] In some embodiments, the method includes administering the Plk1 inhibitor. In some embodiments, the Plk1 inhibitor is BI2536, BI6727, TAK960, NMSP937, GSK461364, or a pharmaceutically acceptable salt thereof.

[0094] In some embodiments, the method includes administering the Uba2 inhibitor. In some embodiments, the Uba2 inhibitor is TAK981 or a pharmaceutically acceptable salt thereof.

[0095] In some embodiments, the method includes administering the Dut inhibitor. In some embodiments, the Dut inhibitor is TAS114 or a pharmaceutically acceptable salt thereof.

[0096] In some embodiments, the method includes administering the Hdac3 inhibitor. In some embodiments, the Hdac3 inhibitor is RGFP966 or a pharmaceutically acceptable salt thereof. ATTORNEY DOCKET NO.: D0963.70010WO00

[0097] In some embodiments, the method includes administering the Chekl inhibitor. In some embodiments, the Chekl inhibitor is SRA737 or a pharmaceutically acceptable salt thereof.

[0098] In some embodiments, the method includes administering the Aurka inhibitor. In some embodiments, the Aurka inhibitor is MLN8237, MK0547, MLN8054, PHA739358, AT9283, AMG900, MK5108, SNS314, TAK901, CYC116, ENMD2076, or a pharmaceutically acceptable salt thereof.

[0099] In some embodiments, the method includes administering the Men1 inhibitor. In some embodiments, the Men1 inhibitor is MI3454, SNDX5613, VTP50469, KO539, or a pharmaceutically acceptable salt thereof.

[0100] In some embodiments, the method includes administering the Doti I inhibitor. In some embodiments, the Doti I inhibitor is EPZ5676 or a pharmaceutically acceptable salt thereof.

[0101] In some embodiments, the method includes administering the Crebbp inhibitor. In some embodiments, the Crebbp inhibitor is CPI4, CCS1477, E7386, NEO1132, NEO2734, PRI724, C82, BC001, C646, EML425, CBP30, or a pharmaceutically acceptable salt thereof.

[0102] In some embodiments, the method includes administering the Ezh2 inhibitor. In some embodiments, the Ezh2 inhibitor is EPZ-6438, GSK126, or a pharmaceutically acceptable salt thereof.

[0103] In some embodiments, the method includes administering the Plk4 inhibitor. In some embodiments, the Plk4 inhibitor is a compound of formula (VI)

[0104]

[0105] or a pharmaceutically acceptable salt thereof,

[0106] wherein

[0107] n is 0, 1, 2, 3, or 4;

[0108] m is 0, 1, or 2;

[0109] L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted C6-10 aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;

[0110] R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;

[0111] R1bis hydrogen; or

[0112] R1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;

[0113] A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted ATTORNEY DOCKET NO.: D0963.70010WO00

[0114] C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl;

[0115] each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl Ci-e alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;

[0116] R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor -N(R3B)2

[0117] each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0118] X is N, and R4is absent; orX is C, and R4is hydrogen, halogen, cyano, optionally substituted amino, optionally substituted acyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;

[0119] R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;

[0120] Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted Ce-io arylene, or optionally substituted C3-8 cycloalkylene;

[0121] R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0122] R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; and R6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

[0123] In some embodiments, the Plk4 inhibitor is a compound of formula (VII)

[0124]

[0125] or a pharmaceutically acceptable salt thereof,

[0126] wherein

[0127] n is 0, 1, 2, 3, or 4;

[0128] m is 0, 1, or 2;

[0129] L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted Ce-io aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;

[0130] R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;

[0131] R1bis hydrogen; or

[0132] R1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;

[0133] A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl;

[0134] each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally ATTORNEY DOCKET NO.: D0963.70010WO00

[0135] substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;

[0136] R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor -N(R3B)2

[0137] each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0138] R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;

[0139] Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted C6-10 arylene, or optionally substituted C3-8 cycloalkylene;

[0140] R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;

[0141] R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; and R6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

[0142] In some embodiments, the Plk4 inhibitor is a compound of formula (VIII)

[0143] R6

[0144]

[0145] (VIII)

[0146] or a pharmaceutically acceptable salt thereof,

[0147] wherein

[0148] n is 0, 1, 2, 3, or 4;

[0149] m is 0, 1, or 2; ATTORNEY DOCKET NO.: D0963.70010WO00

[0150] L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted C6-10 aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;

[0151] R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;

[0152] R1bis hydrogen; or

[0153] R1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;

[0154] A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl;

[0155] each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl Ci-e alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;

[0156] R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor -N(R3B)2

[0157] each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0158] R4is hydrogen, halogen, cyano, optionally substituted amino, optionally substituted acyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0159] R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;

[0160] Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted C6-10 arylene, or optionally substituted C3-8 cycloalkylene;

[0161] R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;

[0162] R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; and R6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

[0163] In some embodiments, the Plk4 inhibitor is selected from the group consisting of compound P1 to P328. In some embodiments, the Plk4 inhibitor is selected from the group consisting of compounds P16, P17, P25, P44, P45, P62, P110, P121, P137, P140, P151, P156, P164, P174, P179, P182, P183, P188, P195, P196, P197, P210, P189, P232, P238, P240, P243, P249, P252, 255, P260, P262, P272, P273, P274, P276, P277, P280, P308, P328, P334, P340, P343, P347, P348, P349, P350, P351, P352, P353, P354, P355, P356, P357, P358, P359, P360, P361, P362, P363, and P364. In some embodiments, the Plk4 inhibitor is centrinone, CFI400945, or a pharmaceutically acceptable salt thereof.

[0164] In some embodiments, the method includes administering the Haspin inhibitor. In some embodiments, the Haspin inhibitor is SEL120 or a pharmaceutically acceptable salt thereof.

[0165] In some embodiments, the method includes administering the Mettl3 inhibitor. In some embodiments, the Mettl3 inhibitor is UZH1a, sTC-15, or a pharmaceutically acceptable salt thereof.

[0166] In some embodiments, the method includes administering the nucleoside analog. In some embodiments, the nucleoside analog is cytarabine, gemcitabine, mercaptopurine, azacytidine, cladribine, decitabine, fluorouracil, floxuridine, fludarabine, nelarabine, or a pharmaceutically acceptable salt thereof or a combination thereof. In some embodiments, the nucleoside analog is gemcitabine or a pharmaceutically acceptable salt thereof. In some embodiments, the nucleoside analog is fluorouracil or a pharmaceutically acceptable salt thereof. In some embodiments, the nucleoside analog is a combination of gemcitabine or a pharmaceutically acceptable salt thereof and fluorouracil or a pharmaceutically acceptable salt thereof.

[0167] In some embodiments, the method includes administering the platinum-based DNA damaging agent. In some embodiments, the platinum-based DNA damaging agent is cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, orsatraplatin. In some embodiments, the platinum-based DNA damaging agent is carboplatin. ATTORNEY DOCKET NO.: D0963.70010WO00

[0168] In some embodiments, the Wee1 inhibitor, Fen1 inhibitor, Topi inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Aurkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod1 inhibitor, Sod2 inhibitor, Bub1 inhibitor, Cdc7 inhibitor, Sae1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Dut inhibitor, Hdac3 inhibitor, Chekl inhibitor, Aurka inhibitor, Men1 inhibitor, Doti I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinum-based DNA damaging agent, or the combination thereof is administered as a pharmaceutical composition.

[0169] In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is a compound of formula (I):

[0170]

[0171] or a pharmaceutically acceptable salt thereof,

[0172] wherein

[0173] each of X, Y, and Z is independently N or CR2;

[0174] R1and each R2are independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, cyano, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or-Q-R7B; or R1combines with one R2that is vicinal to R1to form an optionally substituted C3-6 alkylene;

[0175] each of R3and R4is independently optionally substituted C1-6 alkyl or halogen;

[0176] R5is H or-N(R7)2;

[0177] R6is -C(O)NH(R8), -C(O)R7A, or-SO2R7A;

[0178] each R7is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or-SO2R7A; or two R7groups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0179] each R7Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted C6-10 aryl;

[0180] each R7Bis independently hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -N(R7)2, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or optionally substituted alkoxy;

[0181] each R8is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally ATTORNEY DOCKET NO.: D0963.70010WO00

[0182] substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or two R8, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl; and

[0183] Q is optionally substituted C1-6 alkylene, optionally substituted C2-6 alkenylene, optionally substituted C2-6 alkynylene, optionally substituted C3-8 cycloalkylene, optionally substituted C3-8 cycloalkenylene optionally substituted C6-10 arylene, optionally substituted C2-9 heterocyclylene, or optionally substituted C1-9 heteroarylene.

[0184] In some embodiments, the compound is enriched for the atropisomer of formula (IA):

[0185]

[0186] (IA),

[0187] or a pharmaceutically acceptable salt thereof.

[0188] In some embodiments, X is CR2. In some embodiments, the compound is of formula (II):

[0189]

[0190] or a pharmaceutically acceptable salt thereof.

[0191] In some embodiments, the compound is enriched for the atropisomer of formula (IIA):

[0192]

[0193] or a pharmaceutically acceptable salt thereof.

[0194] In some embodiments, the compound is of formula (III):

[0195]

[0196] ATTORNEY DOCKET NO.: D0963.70010WO00

[0197] (III);

[0198] wherein R2Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or-Q-R7B; or a pharmaceutically acceptable salt thereof.

[0199] In some embodiments, the compound is enriched for the atropisomer of formula (IIIA):

[0200]

[0201] (IIIA);

[0202] or a pharmaceutically acceptable salt thereof.

[0203] In some embodiments, R2Ais hydrogen, optionally substituted C1-6 alkyl, or halogen.

[0204] In some embodiments, R3is optionally substituted C1-6 alkyl. In some embodiments, R3is halogen. In some embodiments, the halogen is chlorine.

[0205] In some embodiments, R4is optionally substituted C1-6 alkyl. In some embodiments, R4is halogen. In some embodiments, the halogen is chlorine. In some embodiments, R2is hydrogen. In some embodiments, R2is optionally substituted C1-6 alkyl. In some embodiments, R2is optionally substituted methyl or optionally substituted isopropyl. In some embodiment, R2is halogen.

[0206] In some embodiments R1is hydrogen. In some embodiments, R1is halogen. In some embodiments, R1is chlorine or bromine. In some embodiments, R1is optionally substituted C1-6 alkyl. In some embodiments, R1is optionally substituted methyl, optionally substituted ethyl, optionally substituted isopropyl, or optionally substituted butyl. In some embodiments, R1is optionally substituted C1-9 heteroaryl. In some embodiments, R1is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, or pyrazolyl, wherein R1is optionally substituted with substituents as defined for optionally substituted C1-9 heteroaryl. In some embodiments, R1is optionally substituted C3-8 cycloalkyl. In some embodiments, R1is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, wherein R1is optionally substituted with substituents as defined for optionally substituted C3-8 cycloalkyl. In some embodiments, R1is optionally substituted C2-9 heterocyclyl. In some embodiments, R1is 1, 2,3,6-tetrahydropyridinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, oxa-aza-spiro[3,3]heptane, or oxa-aza-bicyclo[3.2.1]octane, wherein R1is optionally substituted with substituents as defined for optionally substituted C2-9 heterocyclyl. In some embodiments, R1is optionally substituted C3-8 cycloalkyl. In some embodiments, R1is optionally substituted cyclohexenyl or optionally substituted cyclopentenyl. In some embodiments,

[0207] R1is optionally substituted C6-10 aryl. In some embodiments, R1is optionally substituted phenyl. In some embodiments, R1is -Q-R7B. In some embodiments, Q is optionally substituted C2-6 ATTORNEY DOCKET NO.: D0963.70010WO00

[0208] alkynylene. In some embodiments, Q is optionally substituted C1-6 alkylene. In some embodiments, Q is optionally substituted Ce-io arylene. In some embodiments, R7Bis optionally substituted C2-9 heterocyclyl. In some embodiments, R7Bis optionally substituted Ce-io aryl. In some embodiments, R1is optionally substituted with one, two, or three groups independently selected from the group consisting of methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, oxo, -C(O)NH2, -C(O)NH(Me), -C(O)N(Me)2, -(CH2)n-C(O)OH, and -(CH2)n-C(O)Ot-Bu, wherein n is 0 or 1.

[0209] In some embodiments, R1is -N(R7)2. In some embodiments, R1is diethylamino. In some embodiments,

[0210] In some embodiments, R5is hydrogen. In some embodiments, R5is -N(R7)2.

[0211] In some embodiments, R5is -NH2.

[0212] In some embodiments, R6is -C(O)NH(R8). In some embodiments, R6is -C(O)NH2. In some embodiments, R6is -C(O)NH(Me). In some embodiments, R6is -SO2R7A. In some embodiments, R6is -SO2Me.

[0213] In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is selected from the group consisting of compounds 1-328 and pharmaceutically acceptable salts thereof. In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is administered as a pharmaceutical composition. In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is compound 182 (Table 1), also known as lunresertib.

[0214] In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is compound 182, also known as lunresertib and the method includes administering a Wee1 inhibitor, wherein the WEE1 inhibitor is Debio 0123, also known as zedoresertib.

[0215] In some embodiments of any of the foregoing aspects, the Myt1 inhibitor is compound 182, also known as lunresertib and the method includes administering an ATR inhibitor, wherein the ATR inhibitor is camonsertib.

[0216] Definitions

[0217] The term “aberrant,” as used herein, refers to different from normal. When used to describe activity, aberrant refers to activity that is greater or less than a normal control or the average of normal non-diseased control samples. Aberrant activity may refer to an amount of activity that results in a disease, where returning the aberrant activity to a normal or non-disease-associated amount (e.g., by administering a compound or using a method as described herein), results in reduction of the disease or one or more disease symptoms.

[0218] The term “acyl,” as used herein, represents a group -C(=O)-R, where R is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, or heterocyclyl. Acyl may be optionally substituted as described herein for each respective R group.

[0219] The term “alkanoyl,” as used herein, represents a hydrogen or an alkyl group that is attached to the parent molecular group through a carbonyl group and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, propionyl, butyryl, and iso-butyryl. Unsubstituted alkanoyl groups contain from 1 to 7 carbons. The alkanoyl group may be unsubstituted of substituted (e.g., optionally ATTORNEY DOCKET NO.: D0963.70010WO00

[0220] substituted C1-7 alkanoyl) as described herein for alkyl group. The ending “-oyl” may be added to another group defined herein, e.g., aryl, cycloalkyl, and heterocyclyl, to define “aryloyl,” “cycloalkanoyl,” and “(heterocyclyl)oyl.” These groups represent a carbonyl group substituted by aryl, cycloalkyl, or heterocyclyl, respectively. Each of “aryloyl,” “cycloalkanoyl,” and “(heterocyclyl)oyl” may be optionally substituted as defined for “aryl,” “cycloalkyl,” or “heterocyclyl,” respectively.

[0221] The term “alkenyl,” as used herein, represents acyclic monovalent straight or branched chain hydrocarbon groups of containing one, two, or three carbon-carbon double bonds. Non-limiting examples of the alkenyl groups include ethenyl, prop-1 -enyl, prop-2-enyl, 1 -methylethenyl, but-1-enyl, but-2-enyl, but-3-enyl, 1-methylprop-1-enyl, 2-methylprop-1-enyl, and 1-methylprop-2-enyl. Alkenyl groups may be optionally substituted as defined herein for alkyl.

[0222] The term “alkenylene,” as used herein, refers to a divalent alkenyl group. An optionally substituted alkenylene is an alkenylene that is optionally substituted as described herein for alkenyl.

[0223] The term “alkoxy,” as used herein, represents a chemical substituent of formula -OR, where R is a C1-6 alkyl group, unless otherwise specified. In some embodiments, the alkyl group can be further substituted as defined herein. The term “alkoxy” can be combined with other terms defined herein, e.g., aryl, cycloalkyl, or heterocyclyl, to define an “aryl alkoxy,” “cycloalkyl alkoxy,” and “(heterocyclyl)alkoxy” groups. These groups represent an alkoxy that is substituted by aryl, cycloalkyl, or heterocyclyl, respectively. Each of “aryl alkoxy,” “cycloalkyl alkoxy,” and “(heterocyclyl)alkoxy” may optionally substituted as defined herein for each individual portion.

[0224] The term “alkoxyalkyl,” as used herein, represents a chemical substituent of formula -L-O-R, where L is C1-6 alkylene, and R is C1-6 alkyl. An optionally substituted alkoxyalkyl is an alkoxyalkyl that is optionally substituted as described herein for alkyl.

[0225] The term “alkyl,” as used herein, refers to an acyclic straight or branched chain saturated hydrocarbon group, which, when unsubstituted, has from 1 to 12 carbons, unless otherwise specified. In certain preferred embodiments, unsubstituted alkyl has from 1 to 6 carbons. Alkyl groups are exemplified by methyl; ethyl; n- and iso-propyl; n-, sec-, iso- and tert-butyl; neopentyl, and the like, and may be optionally substituted, valency permitting, with one, two, three, or, in the case of alkyl groups of two carbons or more, four or more substituents independently selected from the group consisting of: amino; alkoxy; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; alkylsulfonyl; alkylsulfinyl; alkylsulfenyl; =O; =S; -C(O)R or-SC>2R, where R is amino; and =NR’, where R’ is H, alkyl, aryl, or heterocyclyl. Each of the substituents may itself be unsubstituted or, valency permitting, substituted with unsubstituted substituent(s) defined herein for each respective group.

[0226] The term “alkylene,” as used herein, refers to a divalent alkyl group. An optionally substituted alkylene is an alkylene that is optionally substituted as described herein for alkyl.

[0227] The term “alkylamino,” as used herein, refers to a group having the formula -N(RN1)2 or-NHRN1, in which RN1is alkyl, as defined herein. The alkyl portion of alkylamino can be optionally substituted as defined for alkyl. Each optional substituent on the substituted alkylamino may itself be unsubstituted or, valency permitting, substituted with unsubstituted substituent(s) defined herein for each respective group. ATTORNEY DOCKET NO.: D0963.70010WO00

[0228] The term “alkylsulfenyl,” as used herein, represents a group of formula —S— (alkyl).

[0229] Alkylsulfenyl may be optionally substituted as defined for alkyl.

[0230] The term “alkylsulfinyl,” as used herein, represents a group of formula -S(O)-(alkyl).

[0231] Alkylsulfinyl may be optionally substituted as defined for alkyl.

[0232] The term “alkylsulfonyl,” as used herein, represents a group of formula -S(O)2-(alkyl).

[0233] Alkylsulfonyl may be optionally substituted as defined for alkyl.

[0234] The term “alkynyl,” as used herein, represents monovalent straight or branched chain hydrocarbon groups of from two to six carbon atoms containing at least one carbon-carbon triple bond and is exemplified by ethynyl, 1-propynyl, and the like. The alkynyl groups may be unsubstituted or substituted (e.g., optionally substituted alkynyl) as defined for alkyl.

[0235] The term “alkynylene,” as used herein, refers to a divalent alkynyl group. An optionally substituted alkynylene is an alkynylene that is optionally substituted as described herein for alkynyl.

[0236] The term “amino,” as used herein, represents -N(RN1)2, where, if amino is unsubstituted, both RN1are H; or, if amino is substituted, each RN1is independently H, -OH, -NO2, -N(RN2)2, -SO2ORN2, -SO2RN2, -SORN2, -C(O)ORN2, an N-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, arylalkyl, aryloxy, cycloalkyl, cycloalkenyl, heteroalkyl, or heterocyclyl, provided that at least one RN1is not H, and where each RN2is independently H, alkyl, or aryl. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) defined herein for each respective group. In some embodiments, amino is unsubstituted amino (i.e., -NH2) or substituted amino (e.g., -NHRN1), where RN1is independently -OH, SO2ORN2, -SO2RN2, -SORN2, -COORN2, optionally substituted alkyl, or optionally substituted aryl, and each RN2can be optionally substituted alkyl or optionally substituted aryl. In some embodiments, substituted amino may be alkylamino, in which the alkyl groups are optionally substituted as described herein for alkyl. In some embodiments, an amino group is -NHRN1, in which RN1is optionally substituted alkyl.

[0237] The term “aryl,” as used herein, represents a mono-, bicyclic, or multicyclic carbocyclic ring system having one or two aromatic rings. Aryl group may include from 6 to 10 carbon atoms. All atoms within an unsubstituted carbocyclic aryl group are carbon atoms. Non-limiting examples of carbocyclic aryl groups include phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, etc. The aryl group may be unsubstituted or substituted with one, two, three, four, or five substituents independently selected from the group consisting of: alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; hydroxy; nitro; thiol; silyl; -(CH2)n-C(O)ORA; -C(O)R; and -SO2R, where R is amino or alkyl, RAis H or alkyl, and n is 0 or 1. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) defined herein for each respective group.

[0238] The term “aryl alkyl,” as used herein, represents an alkyl group substituted with an aryl group. The aryl and alkyl portions may be optionally substituted as the individual groups as described herein.

[0239] The term “arylene,” as used herein, refers to a divalent aryl group. An optionally substituted arylene is an arylene that is optionally substituted as described herein for aryl. ATTORNEY DOCKET NO.: D0963.70010WO00

[0240] The term “aryloxy,” as used herein, represents a chemical substituent of formula -OR, where R is an aryl group, unless otherwise specified. In optionally substituted aryloxy, the aryl group is optionally substituted as described herein for aryl.

[0241] The term “azido,” as used herein, represents an -N3 group.

[0242] The term “biallelic loss-of-function mutation,” as used herein, refers to a mutation that occurs on both alleles of a single gene within a subject’s cell (e.g., cancer cell) that results in the elimination of the active form of a target gene in the cell.

[0243] The term “cancer,” as used herein, refers to all types of cancer, neoplasm or malignant tumors found in mammals (e.g., humans). Exemplary cancers include colorectal cancer (CRC), uterine cancer, bladder cancer, cervical cancer, breast cancer, and gastrointestinal cancer

[0244] The term “carbocyclic,” as used herein, represents an optionally substituted C3-16 monocyclic, bicyclic, or tricyclic structure in which the rings, which may be aromatic or non-aromatic, are formed by carbon atoms. Carbocyclic structures include cycloalkyl, cycloalkenyl, cycloalkynyl, and certain aryl groups.

[0245] The term “carbonyl,” as used herein, represents a -C(O)- group.

[0246] The term “carcinoma,” as used herein, refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases.

[0247] The terms “CCNE1” and “cyclin E1,” as used interchangeably herein, refer to G1 / S specific cyclin E1 (encoded for by the CCNE1 gene). A cell overexpressing CCNE1 is one that exhibits a higher activity of CCNE1 than a cell normally expressing CCNE1. For example, a CCNE1-overexpressing cell is a cell that exhibits a copy number of at least 3 compared to a diploid normal cell with 2 copies. Thus, a cell exhibiting a copy number greater than 3 of CCNE1 is a cell overexpressing CCNE1. The CCNE1 overexpression may be measured by identifying the expression level of the gene product in a cell (e.g., CCNE1 mRNA transcript count or CCNEI protein level).

[0248] The term “cyano,” as used herein, represents -CN group.

[0249] The term “cycloalkenyl,” as used herein, refers to a non-aromatic carbocyclic group having at least one double bond in the ring and from three to ten carbons (e.g., a C3-10 cycloalkenyl), unless otherwise specified. Non-limiting examples of cycloalkenyl include cycloprop-1 -enyl, cycloprop-2-enyl, cyclobut-1-enyl, cyclobut-1-enyl, cyclobut-2-enyl, cyclopent-1 -enyl, cyclopent-2-enyl, cyclopent-3-enyl, norbornen-1-yl, norbornen-2-yl, norbornen-5-yl, and norbornen-7-yl. The cycloalkenyl group may be unsubstituted or substituted (e.g., optionally substituted cycloalkenyl) as described for cycloalkyl.

[0250] The term “cycloalkenylene,” as used herein, represents a divalent cycloalkenyl group. An optionally substituted cycloalkenylene is a cycloalkenylene that is optionally substituted as described herein for cycloalkyl.

[0251] The term “cycloalkoxy,” as used herein, represents a chemical substituent of formula -OR, where R is cycloalkyl group, unless otherwise specified. In some embodiments, the cycloalkyl group can be further substituted as defined herein.

[0252] The term “cycloalkyl,” as used herein, refers to a cyclic alkyl group having from three to ten carbons (e.g., a C3-C10 cycloalkyl), unless otherwise specified. Cycloalkyl groups may be monocyclic or bicyclic. Bicyclic cycloalkyl groups may be of bicyclo[p.q. O]alkyl type, in which each of p and q is, ATTORNEY DOCKET NO.: D0963.70010WO00

[0253] independently, 1, 2, 3, 4, 5, 6, or 7, provided that the sum of p and q is 2, 3, 4, 5, 6, 7, or 8.

[0254] Alternatively, bicyclic cycloalkyl groups may include bridged cycloalkyl structures, e.g., bicyclo[p.q.r]alkyl, in which r is 1, 2, or 3, each of p and q is, independently, 1, 2, 3, 4, 5, or 6, provided that the sum of p, q, and r is 3, 4, 5, 6, 7, or 8. The cycloalkyl group may be a spirocyclic group, e.g., spiro[p.q]alkyl, in which each of p and q is, independently, 2, 3, 4, 5, 6, or 7, provided that the sum of p and q is 4, 5, 6, 7, 8, or 9. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 1-bicyclo[2.2.1 Jheptyl, 2-bicyclo[2.2.1 Jheptyl, 5-bicyclo[2.2.1 Jheptyl, 7-bicyclo[2.2.1 Jheptyl, and decalinyl. The cycloalkyl group may be unsubstituted or substituted (e.g., optionally substituted cycloalkyl) with one, two, three, four, or five substituents independently selected from the group consisting of: alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; =O; =S; -SO2R, where R is optionally substituted amino; =NR’, where R’ is H, alkyl, aryl, or heterocyclyl; and -CON(RA)2, where each RAis independently H or alkyl, or both RA, together with the atom to which they are attached, combine to form heterocyclyl. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) defined herein for each respective group.

[0255] The term “cycloalkylene,” as used herein, represents a divalent cycloalkyl group. An optionally substituted cycloalkylene is a cycloalkylene that is optionally substituted as described herein for cycloalkyl.

[0256] The term “cycloalkynyl,” as used herein, refers to a monovalent carbocyclic group having one or two carbon-carbon triple bonds and having from eight to twelve carbons, unless otherwise specified. Cycloalkynyl may include one transannular bond or bridge. Non-limiting examples of cycloalkynyl include cyclooctynyl, cyclononynyl, cyclodecynyl, and cyclodecadiynyl. The cycloalkynyl group may be unsubstituted or substituted (e.g., optionally substituted cycloalkynyl) as defined for cycloalkyl.

[0257] “Disease” or “condition” refer to a state of being or health status of a patient or subject capable of being treated with the compounds or methods provided herein.

[0258] The term “FBXW7,” as used herein, refers to F-box / WD Repeat-Containing Protein 7 transcript (gene name: FBXW7). An FBXW7 gene having an inactivation mutation, is one that fails to produce a functional FBXW7 protein or produces reduced quantities of FBXW7 protein in a cell.

[0259] The term “halo,” as used herein, represents a halogen selected from bromine, chlorine, iodine, and fluorine.

[0260] The term “heteroalkyl,” as used herein refers to an alkyl, alkenyl, or alkynyl group interrupted once by one or two heteroatoms; twice, each time, independently, by one or two heteroatoms; three times, each time, independently, by one or two heteroatoms; or four times, each time, independently, by one or two heteroatoms. Each heteroatom is, independently, O, N, or S. In some embodiments, the heteroatom is O or N. None of the heteroalkyl groups includes two contiguous oxygen or sulfur atoms. The heteroalkyl group may be unsubstituted or substituted (e.g., optionally substituted heteroalkyl). When heteroalkyl is substituted and the substituent is bonded to the heteroatom, the substituent is ATTORNEY DOCKET NO.: D0963.70010WO00

[0261] selected according to the nature and valency of the heteroatom. Thus, the substituent bonded to the heteroatom, valency permitting, is selected from the group consisting of =O, -N(RN2)2, -SO2ORN3, -SO2RN2, -SORN3, -COORN3, an N protecting group, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, or cyano, where each RN2is independently H, alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, or heterocyclyl, and each RN3is independently alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, or heterocyclyl. Each of these substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) defined herein for each respective group. When heteroalkyl is substituted and the substituent is bonded to carbon, the substituent is selected from those described for alkyl, provided that the substituent on the carbon atom bonded to the heteroatom is not Cl, Br, or I. It is understood that carbon atoms are found at the termini of a heteroalkyl group.

[0262] The term “heteroarylene,” as used herein, represents a divalent heteroaryl. An optionally substituted heteroarylene is a heteroarylene that is optionally substituted as described herein for heteroaryl.

[0263] The term “heterocyclyl,” as used herein, represents a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having fused, bridging, and / or spiro 3-, 4-, 5-, 6-, 7-, or 8-membered rings, unless otherwise specified, containing one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. In some embodiments, “heterocyclyl” is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having fused or bridging 5-, 6-, 7-, or 8-membered rings, unless otherwise specified, containing one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocyclyl can be aromatic or non-aromatic. Non-aromatic 5-membered heterocyclyl has zero or one double bonds, non-aromatic 6- and 7-membered heterocyclyl groups have zero to two double bonds, and non-aromatic 8-membered heterocyclyl groups have zero to two double bonds and / or zero or one carboncarbon triple bond. Heterocyclyl groups include from 1 to 16 carbon atoms unless otherwise specified. Certain heterocyclyl groups may include up to 9 carbon atoms. Non-aromatic heterocyclyl groups include pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, homopiperidinyl, piperazinyl, pyridazinyl, oxazolidinyl, isoxazolidiniyl, morpholinyl, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, thiazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, dihydroindolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, pyranyl, dihydropyranyl, dithiazolyl, etc. If the heterocyclic ring system has at least one aromatic resonance structure or at least one aromatic tautomer, such structure is an aromatic heterocyclyl (i.e., heteroaryl). Non-limiting examples of heteroaryl groups include benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, furyl, imidazolyl, indolyl, isoindazolyl, isoquinolinyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl, pyrrolyl, pyridinyl, pyrazinyl, pyrimidinyl, qunazolinyl, quinolinyl, thiadiazolyl (e.g., 1,3,4-thiadiazole), thiazolyl, thienyl, triazolyl, tetrazolyl, etc. The term “heterocyclyl” also represents a heterocyclic compound having a bridged multicyclic structure in which one or more carbons and / or heteroatoms bridges two non-adjacent members of a monocyclic ring, e.g., quinuclidine, tropanes, ordiaza-bicyclo[2.2.2]octane. The term “heterocyclyl” includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings is fused to one, two, or ATTORNEY DOCKET NO.: D0963.70010WO00

[0264] three carbocyclic rings, e.g., an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocyclic ring. Examples of fused heterocyclyls include 1,2,3,5,8,8a-hexahydroindolizine; 2,3-dihydrobenzofuran; 2,3-dihydroindole; and 2,3-dihydrobenzothiophene. The heterocyclyl group may be unsubstituted or substituted with one, two, three, four or five substituents independently selected from the group consisting of: alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; hydroxy; nitro; thiol; silyl; cyano; -C(O)R or -SO2R, where R is amino or alkyl; =O; =S; =NR’, where R’ is H, alkyl, aryl, or heterocyclyl. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) defined herein for each respective group.

[0265] The term “heterocyclylene,” as used herein, represents a divalent heterocyclyl. An optionally substituted heterocyclylene is a heterocyclylene that is optionally substituted as described herein for heterocyclyl.

[0266] The term “(heterocyclyl)oxy,” as used herein, represents a chemical substituent of formula -OR, where R is a heterocyclyl group, unless otherwise specified. (Heterocyclyl)oxy can be optionally substituted in a manner described for heterocyclyl.

[0267] The terms “hydroxyl” and “hydroxy,” as used interchangeably herein, represent an -OH group. The term “inactivating mutation,” as used herein, refers to a mutation to a gene that results in the gene encoding an inactive form of a protein (e.g., a protein which cannot perform the normal cellular functions of said protein). Exemplary inactivating mutations are truncating mutations, missense mutations (e.g., missense mutations encoding an amino acid substitution selected from R465, R479, and / or R505), and truncating biallelic loss-of-function mutations.

[0268] The term “isotopically enriched,” as used herein, refers to the pharmaceutically active agent with the isotopic content for one isotope at a predetermined position within a molecule that is at least 100 times greater than the natural abundance of this isotope. For example, a composition that is isotopically enriched for deuterium includes an active agent with at least one hydrogen atom position having at least 100 times greater abundance of deuterium than the natural abundance of deuterium. Preferably, an isotopic enrichment for deuterium is at least 1000 times greater than the natural abundance of deuterium. More preferably, an isotopic enrichment for deuterium is at least 4000 times greater (e.g., at least 4750 times greater, e.g., up to 5000 times greater) than the natural abundance of deuterium.

[0269] The term “Myt1,” as used herein, refers to membrane-associated tyrosine and threoninespecific CDC2-inhibitory kinase (Myt1) (Gene name PKMYT1).

[0270] The term “Myt1 inhibitor,” as used herein, represents a compound that upon contacting the enzyme Myt1, whether in vitro, in cell culture, or in an animal, reduces the activity of Myt1, such that the measured Myt1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Myt1 inhibitors, the Myt1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 3 nM or less) and could be as low as 100 pM or 10 pM. Preferably, the Myt1 IC50 is 1 nM to 1 pM (e.g., 1 nM to 750 nM, 1 nM to 500 nM, or 1 nM to 250 nM). Even more preferably, the Myt1 IC50 is less than 20 nm (e.g., 1 nM to 20 nM).

[0271] The term “nitro,” as used herein, represents an -NO2 group. ATTORNEY DOCKET NO.: D0963.70010WO00

[0272] The term “oxo,” as used herein, represents a divalent oxygen atom (e.g., the structure of oxo may be shown as =O).

[0273] The term “Ph,” as used herein, represents phenyl.

[0274] The term “pharmaceutical composition,” as used herein, represents a composition containing a compound described herein, formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein.

[0275] The term “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier,” as used interchangeably herein, refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving the active compound) and having the properties of being nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0276] The term “pharmaceutically acceptable salt,” as use herein, represents those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. PH. Stahl and C. G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, ATTORNEY DOCKET NO.: D0963.70010WO00

[0277] malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like.

[0278] The term “subject,” as used herein, represents a human or non-human animal (e.g., a mammal) that is suffering from, or is at risk of, disease or condition, as determined by a qualified professional (e.g., a doctor or a nurse practitioner) with or without known in the art laboratory test(s) of sample(s) from the subject. Preferably, the subject is a human. Non-limiting examples of diseases and conditions include diseases having the symptom of cell hyperproliferation, e.g., a cancer.

[0279] The term “tautomer” refers to structural isomers that readily interconvert, often by relocation of a proton. Tautomers are distinct chemical species that can be identified by differing spectroscopic characteristics, but generally cannot be isolated individually. Non-limiting examples of tautomers include ketone - enol, enamine - imine, amide - imidic acid, nitroso - oxime, ketene - ynol, and amino acid - ammonium carboxylate.

[0280] “Treatment” and “treating,” as used herein, refer to the medical management of a subject with the intent to improve, ameliorate, stabilize, prevent, or cure a disease or condition. This term includes active treatment (treatment directed to improve the disease or condition); causal treatment (treatment directed to the cause of the associated disease or condition); palliative treatment (treatment designed for the relief of symptoms of the disease or condition); preventative treatment (treatment directed to minimizing or partially or completely inhibiting the development of the associated disease or condition); and supportive treatment (treatment employed to supplement another therapy).

[0281] The term “truncating biallelic loss-of-function mutation,” as used herein, refers to a biallelic mutation within a subject’s cell (e.g., cancer cell) that results in the elimination of an active form of a target gene (e.g., FBXW7) in the cell, wherein the biallelic mutation includes (i) at least one truncating mutation (e.g., a nonsense, nonstop, frameshift, or splice site mutation) in a first allele of the target gene (e.g., FBXW7), and (ii) another truncating mutation ora missense mutation (e.g., missense mutations encoding an amino acid substitution selected from R465, R479, and / or R505) in a second allele of the target gene (e.g., FBXW7), or a partial or complete deletion (e.g., a loss of heterozygosity) of the second allele of the target gene (e.g., FBXW7).

[0282] As used herein, the term “truncating mutation” refers to (i) any mutation (e.g., a nonsense mutation, a nonstop mutation, a frameshift mutation (e.g., resulting from an insertion or deletion), ora splice site mutation) in a gene that results in a shorter gene product (e.g., mRNAor polypeptide) relative to wild type, or (ii) a partial or complete deletion of the gene. ATTORNEY DOCKET NO.: D0963.70010WO00

[0283] BRIEF DESCRIPTION OF THE DRAWINGS FIG 1A is a stacked bar chart showing Cyclin E over-expression (e.g., H-Score >100) measured by immunohistochemistry (IHC) in biobank tissue samples unselected for lunresertib biomarkers.

[0284] FIG 1B is a table showing Cyclin E over-expression (e.g., H-Score >100) measured by immunohistochemistry (IHC) in baseline tissue samples collected from patients enrolled in National Clinical Trial (NCT) number: NCT04855656 (“MYTHIC”). Taken together with FIG. 1A, these data illustrate that selecting for lunresertib biomarkers, such as F-box / WD repeat containing protein (FBXW7) in colorectal cancer (CRC), can enrich for patients having Cyclin E protein over-expression.

[0285] FIG. 2 is a lollipop graph showing the frequency of mutations across the FBXW7 protein in patients enrolled in MYTHIC. This graph highlights three common arginine (R) hotspot mutations (R465, R479, and R505) within the third and fourth WD40 repeats that disrupt recognition of the Cyclin E1 substrate and are classified as deleterious, missense mutations. Also indicated are several truncating, inframe, and splice site mutations that span FBXW7.

[0286] FIG 3 provides a set of box plots showing Cyclin E protein expression stratified by groups of FBXW7 loss-of-fu notion (LoF) quality in non-gynecological (left) and CRC (right) patients enrolled in MYTHIC. “High” confidence FBXW7 mutations include hotspots mutations (R465, R479, R505) and biallelic truncations. “Low” FBXW7 mutations include monoallelic truncations, missense, and splice mutations. Non-hotspot mutations where allelic status is indeterminate, or biallelic missense and splice mutations are classified as “unknown.” Non-gynecological cancers include all indications except for ovarian and endometrial cancers. These data illustrate that hotspots mutations (R465, R479, R505) and biallelic truncation mutations result in high Cyclin E accumulation.

[0287] FIG 4 is a graph showing the duration of on-treatment for CRC patients treated with lunresertib (RP-6306) in combination with camonsertib (RP-3500) in MYTHIC, stratified by FBXW7 LoF quality. High and low Cyclin E protein expression levels measured by IHC are annotated.

[0288] FIG 5A is a western blot image showing increased Cyclin E1 accumulation in retinal RPE1 cell lines following FBXW7 truncation by clustered regularly interspaced short palindromic repeats (CRISPR)-mediated base editing (CRISPR-BE), relative to control guide (sgPLK1silent). Biallelic status of these truncations were independently confirmed (data not shown).

[0289] FIG. 5B is a quantification of the western blot in FIG. 5A.

[0290] FIG. 5C is a growth curve showing that FBXW7 truncations in RPE1 cell lines increased their sensitivity to lunresertib (RP-6306) treatment. Biallelic status of these truncations were independently confirmed (data not shown).

[0291] FIG 5D is a western blot image showing increased Cyclin E1 accumulation in colorectal DLD1 cell lines following FBXW7 truncation by CRISPR-BE, relative to control guide (sgPLK1silent). Biallelic status of these truncations were independently confirmed (data not shown).

[0292] FIG. 5E is a quantification of the western blot in FIG. 5D.

[0293] FIG. 5F is a growth curve showing that FBXW7 truncations in DLD1 cell lines increased their sensitivity to lunresertib (RP-6306) treatment. Biallelic status of these truncations were independently confirmed (data not shown). ATTORNEY DOCKET NO.: D0963.70010WO00

[0294] FIG. 6 is a schematic illustrating a working hypothesis of the effect of FBXW7 LoF quality on Cyclin E accumulation. As illustrated, only hotspot FBXW7 missense and biallelic truncating mutations cause true LoF. WT = wild type; PTC = premature termination codon; NMD = nonsense-mediated decay; LOH = loss-of-heterozygosity.

[0295] DETAILED DESCRIPTION

[0296] In general, the present disclosure relates to the discovery of the susceptibility of cells (e.g., cancer cells, e.g., tumors) harboring mutations in genes which encode for F-box proteins (e.g., FBXW7, which encodes F-box / WD repeat containing protein (FBXW7)) to treatment with a membrane-associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitors. For example, the present disclosure highlights that the treatment of cancer cells having specific mutations in FBXW7 can increase the cells sensitivity to treatment with Myt1 inhibitors. Advantageously, Applicant discovered that cancer cells containing truncating biallelic loss-of-function mutations in FBXW7 are particularly susceptible to Myt1 inhibitors. Exemplary truncating biallelic loss-of-function mutations are: (i) a truncating mutation on a first FBXW7 allele and a missense mutation (e.g., hotspot mutation) on a second FBXW7 allele; (ii) a truncating mutation on a first FBXW7 allele and a truncating mutation on a second FBXW7 allele; (iii) a truncating mutation on a first allele and a partial or complete deletion of the second FBXW7 allele.

[0297] The methods described herein are useful for the treatment of cells (e.g., cancer cells) having, for example, a truncating biallelic loss-of-function mutation in FBXW7. Such cells may be treated by a drug (e.g., an Myt1 inhibitor, e.g., any one of compounds 1 to 328) or combination of drugs described herein. Accordingly, cancers harboring a FBXW7 mutation may be particularly susceptible to treatment with a Myt1 inhibitor (e.g., any one of compounds 1 to 328 described herein). Methods of identifying truncating biallelic loss-of-function mutations and treating cancer in a subject with a Myt1 inhibitor (e.g., as a mono- or combinatorial therapy) are described further below.

[0298] Truncating Biallelic Loss-of-function Mutations

[0299] The methods described herein may be utilized on cells (e.g., cancer cells) that have or are suspected of having a truncating biallelic loss-of-function mutation in a gene encoding an F-box protein. A truncating biallelic loss-of-function mutation is a biallelic mutation (e.g., both alleles of a single gene are mutated) that produces a loss-of-function (e.g., an elimination of an active form of a target gene). The biallelic mutation includes a truncating mutation (e.g., a nonsense, nonstop, frameshift, or splice site mutation) on one allele. The biallelic mutation may further include a missense (e.g., non-synonymous or non-conservative) mutation, another truncating mutation on the other allele of the same gene, or loss-of-heterozygosity of the gene. For example, a truncating biallelic loss-of-function mutation in FBXW7 may include a truncating mutation on a first FBXW7 allele and a missense (e.g., non-conservative) mutation on a second FBXW7 allele in a subject’s cell (e.g., a cancer cell). In another example, a truncating biallelic loss-of-function mutation in FBXW7 may include a truncating mutation on a first FBXW7 allele and a truncating mutation on a second FBXW7 allele in a subject’s cell (e.g., a cancer cell). In yet another example, a truncating biallelic loss-of- ATTORNEY DOCKET NO.: D0963.70010WO00

[0300] function mutation in FBXW7 may include a truncating mutation on the first FBXW7 allele and a deletion of the second allele (referred to as loss-of-heterozygosity) in a subject’s cell. In yet another example, a truncating biallelic loss-of-function mutation in FBXW7 may include a truncating mutation on the first FBXW7 allele and a partial or complete deletion of the second allele in a subject’s cell.

[0301] Truncating mutations may produce a premature stop codon in the coding region of a gene or splice variant of a gene, resulting in a gene product (e.g., an mRNA and / or a polypeptide) that is shorter than wild type. A shorter gene product (e.g. an mRNA and / or a polypeptide) may lose some or all its function and / or be degraded by the cell, thereby producing a loss-of-function of the gene and its products. Exemplary stop codons include UAG, UAA, and UGA (in the context of mRNA) or TAG, TAA, and TGA (in the context of DNA).

[0302] Truncating mutations of a truncating biallelic loss-of-function mutation may produce a gene product (e.g., an mRNA or a polypeptide) that is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% shorter than a wild type gene product. For example, a truncating mutation in FBXW7 may produce an mRNA or polypeptide sequence that is at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% shorter than a wild type FBXW7 mRNA or polypeptide sequence.

[0303] In another example, a truncating mutation in FBXW7 may produce an mRNA sequence that is less than 10 nucleotides, less than 50 nucleotides, less than 100 nucleotides, less than 150 nucleotides, less than 200 nucleotides, less than 250 nucleotides, less than 300 nucleotides, less than 350 nucleotides, less than 400 nucleotides, less than 450 nucleotides, less than 500 nucleotides, less than 550 nucleotides, less than 600 nucleotides, less than 650 nucleotides, less than 700 nucleotides, less than 750 nucleotides, less than 800 nucleotides, less than 850 nucleotides, less than 900 nucleotides, less than 950 nucleotides, less than 1000 nucleotides, less than 1050 nucleotides, less than 1100 nucleotides, less than 1150 nucleotides, less than 1200 nucleotides, less than 1250 nucleotides, less than 1300 nucleotides, less than 1350 nucleotides, less than 1400 nucleotides, less than 1450 nucleotides, less than 1500 nucleotides, less than 1550 nucleotides, less than 1600 nucleotides, less than 1650 nucleotides, less than 1700 nucleotides, less than 1750 nucleotides, less than 1800 nucleotides, less than 1850 nucleotides, less than 1900 nucleotides, or less than 1950 nucleotides in length.

[0304] In another example, a truncating mutation in FBXW7 may produce an mRNA sequence that is 50-1900 nucleotides, 50-1800 nucleotides, 50-1700 nucleotides, 50-1600 nucleotides, 50-1500 nucleotides, 50-1400 nucleotides, 50-1300 nucleotides, 50-1200 nucleotides, 100-1850 nucleotides, 150-1800 nucleotides, 200-1750 nucleotides, 250-1700 nucleotides, 300-1650 nucleotides, 350-1600 nucleotides, 400-1550 nucleotides, 450-1500 nucleotides, 500-1450 nucleotides, 550-1400 nucleotides, 600-1350 nucleotides, 650-1300 nucleotides, or 700-1250 nucleotides in length.

[0305] In another example, a truncating mutation in FBXW7 may produce a polypeptide sequence that is less than 50 amino acids, less than 100 amino acids, less than 150 amino acids, less than 200 amino acids, less than 250 amino acids, less than 300 amino acids, less than 350 amino acids, less than 400 amino acids, less than 450 amino acids, less than 500 amino acids, less than 550 amino acids, less than 600 amino acids, less than 650 amino acids, or less than 700 amino acids in length. ATTORNEY DOCKET NO.: D0963.70010WO00

[0306] In another example, a truncating mutation in FBXW7 may produce a protein sequence that is 50-600 amino acids, 50-550 amino acids, 50-500 amino acids, 50-450 amino acids, 50-400 amino acids, 50-350 amino acids, 50-300 amino acids, 50-250 amino acids, 50-200 amino acids, 50-150 amino acids, 50-100 amino acids, 100-600 amino acids, 150-550 amino acids, 200-500 amino acids, 250-450 amino acids, or 300-400 amino acids in length.

[0307] Non-limiting examples of truncating mutations in FBXW7 may lead to truncations at positions p. Q306*, p. R393*, p. R479*, p. S546* and / or p. W486* of an FBXW7 polypeptide.

[0308] Missense mutations change the nucleotide sequence of a codon such that the codon encodes a different amino acid. For example, the codon “GTG” encodes the amino acid valine. A missense mutation changing the thymine (T) to a cytosine

[0309]

[0310] (C) (e.g., “GTG” “GCG”) would instead encode the amino acid alanine. Missense mutation may lose some or all of its function and / or be degraded by the cell, thereby producing a loss-of-function of the gene. Missense mutations of a truncating biallelic loss-of-function mutation may occur anywhere along the coding region of a gene. In some embodiments of the invention, the missense mutation is at a codon that encodes an arginine (R). Exemplary arginine codons include CGU, CGC, CGA, CGG, AGA, and AGG (in the context of mRNA) or CGT, CGC, CGA, CGG, AGA, and AGG (in the context of DNA). In some embodiments of the invention, at least one, at least two, or three missense mutations encodes for an amino acid substitution at R465, R479, and / or R505 of an FBXW7 polypeptide.

[0311] In other embodiments of the invention, the missense mutation is at a codon that encodes an alanine (A). In other embodiments of the invention, the missense mutation is at a codon that encodes an asparagine (N). In other embodiments of the invention, the missense mutation is at a codon that encodes an aspartate (D). In other embodiments of the invention, the missense mutation is at a codon that encodes a cysteine (C). In other embodiments of the invention, the missense mutation is at a codon that encodes a glutamine (Q). In other embodiments of the invention, the missense mutation is at a codon that encodes a glutamate (E). In other embodiments of the invention, the missense mutation is at a codon that encodes a glycine (G). In other embodiments of the invention, the missense mutation is at a codon that encodes a histidine (H). In other embodiments of the invention, the missense mutation is at a codon that encodes an isoleucine (I). In other embodiments of the invention, the missense mutation is at a codon that encodes a leucine (L). In other embodiments of the invention, the missense mutation is at a codon that encodes a lysine (K). In other embodiments of the invention, the missense mutation is at a codon that encodes a methionine (M). In other embodiments of the invention, the missense mutation is at a codon that encodes a phenylalanine (F). In other embodiments of the invention, the missense mutation is at a codon that encodes a proline (P). In other embodiments of the invention, the missense mutation is at a codon that encodes a serine (S). In other embodiments of the invention, the missense mutation is at a codon that encodes a threonine (T). In other embodiments of the invention, the missense mutation is at a codon that encodes a tryptophan (W). In other embodiments of the invention, the missense mutation is at a codon that encodes a tyrosine (Y). In other embodiments of the invention, the missense mutation is at a codon that encodes a valine (V). ATTORNEY DOCKET NO.: D0963.70010WO00

[0312] Identification of Mutations

[0313] The methods of the present disclosure may include the step of identifying a cell (e.g., a cancer cell) as having a truncating biallelic loss-of-function mutation in an F-box-encoding gene (e.g., FBXW7). Any sequencing technique known in the field may be used to identify a cell as having a truncating biallelic loss-of-function mutation in an F-box-encoding gene (e.g., FBXW7). For example, identifying a cancer cell as having a truncating biallelic loss-of-function mutation in FBXW7 can include any next generation sequencing (NGS) technique known in the art, such as targeted panel sequencing, whole genome sequencing (WGS) and Whole Exome Sequencing (WES).

[0314] By way of example, identifying a cancer cell as having a truncating biallelic loss-of-function mutation in FBXW7 may include a step of determining from read counts for a plurality of single nucleotide variants (SNVs) including homozygous and heterozygous SNVs obtained from sequencing a sample including the cancer cell and from reference read counts and determining an integer total copy number of a locus segment within a target gene (e.g., FBXW7) region in a cancer cell from the subject or in the cancer cell and / or two integer allele-specific copy numbers of the locus segment.

[0315] The cancer may then be identified as having a truncating biallelic loss-of-function mutation (e.g., in FBXW7) if it has at least one truncation mutation described as introduction of a stop codon, or shift in protein frame and one of the following: (a) the integer total copy number is > 0, and the integer allele-specified copy number is 0 (e.g., at the locus where the FBXW7-inactivating mutation is found) leading to loss-of-heterozygosity; or (b) a mutation of any type (e.g., missense, truncation, or splice-sire mutation) in the other allele of FBXW7. When the integer total copy number and the integer allelespecific copy numbers are 0, the detected mutation is a homozygous deletion and will also be considered a truncating biallelic loss-of-function.

[0316] For example, the step of determining allelic status may include: from read counts for the plurality of SNVs including homozygous and heterozygous SNVs obtained from sequencing a sample including the cancer cell and from reference read counts, determining total copy number log-ratios, allelic copy number log-odds ratios, and target coverage values for the heterozygous SNVs; segmenting the total copy number log-ratios and the allelic copy number log-odds ratios; estimating sample purity and sample ploidy for the cancer cell from the total copy number log-ratios and the target coverage values; and from the target coverage values, the sample purity, the sample ploidy, the total copy number log-ratios, and the allelic copy number log-odds ratios, generating an integer total copy number of a segment including a plurality of heterozygous single nucleotide variants (SNVs) within the FBXW7 gene region in the cancer cell and two integer allele-specific copy numbers of the segment. Typically, the cell from the subject is provided as a biopsy. Read counts may be obtained using next generation sequencing of the cells in the sample with or without a panel of normal samples.

[0317] Additional methods for identifying cancers as having a biallelic loss-of-function mutation are known in the art (see, e.g., WO2018 / 039463, WO2019 / 020652, and WO2022 / 226655, the methods of which are incorporated herein by reference). ATTORNEY DOCKET NO.: D0963.70010WO00

[0318] Myt1 Inhibitors

[0319] The compound used in the methods of the invention may be, e.g., a compound of formula (I):

[0320]

[0321] or a pharmaceutically acceptable salt thereof,

[0322] wherein:

[0323] each of X, Y, and Z is independently N or CR2;

[0324] R1and each R2are independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, cyano, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or-Q-R7B; or R1combines with one R2that is vicinal to R1to form an optionally substituted C3-6 alkylene;

[0325] each of R3and R4is independently optionally substituted C1-6 alkyl or halogen;

[0326] R5is H or-N(R7)2;

[0327] R6is -C(O)NH(R8), -C(O)R7A, or-SO2R7A;

[0328] each R7is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or-SO2R7A; or two R7groups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0329] each R7Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted C6-10 aryl;

[0330] each R7Bis independently hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -N(R7)2, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or optionally substituted alkoxy;

[0331] each R8is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or two R8, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl; and

[0332] Q is optionally substituted C1-6 alkylene, optionally substituted C2-6 alkenylene, optionally substituted C2-6 alkynylene, optionally substituted C3-8 cycloalkylene, optionally substituted C3-8 cycloalkenylene optionally substituted C6-10 arylene, optionally substituted C2-9 heterocyclylene, or optionally substituted C1-9 heteroarylene. ATTORNEY DOCKET NO.: D0963.70010WO00

[0333] Preferably, the compound of formula (I) is enriched for the atropisomer of formula (IA):

[0334]

[0335] (IA);

[0336] where all variables are as described herein.

[0337] The compound used in the methods of the invention may be, e.g., a compound of formula (II):

[0338]

[0339] where all variables are as described herein.

[0340] Preferably, the compound of formula (II) is enriched for the atropisomer of formula (IIA):

[0341]

[0342] (HA);

[0343] where all variables are as described herein.

[0344] The compound used in the methods of the invention may be, e.g., a compound of formula (HI):

[0345]

[0346] (HI);

[0347] where R2Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, ATTORNEY DOCKET NO.: D0963.70010WO00

[0348] optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or-Q-R7BPreferably, the compound of formula (III) is enriched for the atropisomer of formula (IIIA):

[0349]

[0350] (IIIA);

[0351] The compound used in the methods of the invention may be, e.g., a compound listed in Table 1 below or a pharmaceutically acceptable salt thereof.

[0352] Table 1

[0353]

[0354] ATTORNEY DOCKET NO.: D0963.70010WO00

[0355]

[0356] ATTORNEY DOCKET NO.: D0963.70010WO00

[0357]

[0358] ATTORNEY DOCKET NO.: D0963.70010WO00

[0359]

[0360] ATTORNEY DOCKET NO.: D0963.70010WO00

[0361]

[0362] ATTORNEY DOCKET NO.: D0963.70010WO00

[0363]

[0364] ATTORNEY DOCKET NO.: D0963.70010WO00

[0365]

[0366] ATTORNEY DOCKET NO.: D0963.70010WO00

[0367]

[0368] ATTORNEY DOCKET NO.: D0963.70010WO00

[0369]

[0370] ATTORNEY DOCKET NO.: D0963.70010WO00 104 105 106

[0371] OH H2NH2N OH H2NH^CI> A H2NH2NCAZN'Y^OH

[0372] NZ XN N N NZ XN

[0373] 107 110 111

[0374] OH OH OH

[0375] H2N NH2VA H2NH2N \X. H2NH^N

[0376] NZ XN N N N N CD

[0377] ^~^CD3 3r

[0378] 112 113 114

[0379] H2NH2^ OH H2NH2N D3C_^ H2NH2ND3C\^4

[0380] O^ ^ OH O'^ V? S3TS< OHNZ XN NH

[0381] <^~ ^CD3 / \D3C

[0382] 115 116 117

[0383] OH OH OH

[0384] H2NH2N D3C^L H2N YH2N

[0385] 0H2H2J^ °^A_ / ND3C AA--M

[0386] NH

[0387] 118 119 120

[0388] H2NH2N OH OH i X Z^z H2N

[0389] ZNNH. V^H2NH2N X " OH

[0390] / ~Al

[0391]

[0392] ATTORNEY DOCKET NO.: D0963.70010WO00

[0393]

[0394] ATTORNEY DOCKET NO.: D0963.70010WO00

[0395]

[0396] ATTORNEY DOCKET NO.: D0963.70010WO00

[0397]

[0398] ATTORNEY DOCKET NO.: D0963.70010WO00

[0399]

[0400] ATTORNEY DOCKET NO.: D0963.70010WO00

[0401]

[0402] ATTORNEY DOCKET NO.: D0963.70010WO00

[0403]

[0404] ATTORNEY DOCKET NO.: D0963.70010WO00

[0405]

[0406] ATTORNEY DOCKET NO.: D0963.70010WO00

[0407]

[0408] ATTORNEY DOCKET NO.: D0963.70010WO00

[0409]

[0410] ATTORNEY DOCKET NO.: D0963.70010WO00

[0411]

[0412] ATTORNEY DOCKET NO.: D0963.70010WO00

[0413]

[0414] ATTORNEY DOCKET NO.: D0963.70010WO00

[0415]

[0416] ATTORNEY DOCKET NO.: D0963.70010WO00

[0417]

[0418] ATTORNEY DOCKET NO.: D0963.70010WO00

[0419]

[0420] ATTORNEY DOCKET NO.: D0963.70010WO00

[0421]

[0422] ATTORNEY DOCKET NO.: D0963.70010WO00

[0423]

[0424] The methods of the invention include (where possible) individual diastereomers, enantiomers, epimers, and atropisomers of the compounds disclosed herein, and mixtures of diastereomers and / or enantiomers thereof including racemic mixtures. Although the specific stereochemistries disclosed herein are preferred, other stereoisomers, including diastereomers, enantiomers, epimers, atropisomers, and mixtures of these may also have utility in treating Myt1 -mediated diseases. Inactive or less active diastereoisomers and enantiomers may be useful, e.g., for scientific studies relating to the receptor and the mechanism of activation.

[0425] It is understood that certain molecules can exist in multiple tautomeric forms. This invention includes all tautomers even though only one tautomer may be indicated in the examples.

[0426] The invention also includes pharmaceutically acceptable salts of the compounds, and pharmaceutical compositions that contain the compounds and a pharmaceutically acceptable carrier. The compounds are especially useful, e.g., in certain kinds of cancer and for slowing the progression of cancer once it has developed in a patient.

[0427] The compounds disclosed herein may be used in pharmaceutical compositions including (a) the compound(s) or pharmaceutically acceptable salts thereof, and (b) a pharmaceutically acceptable carrier. The compounds may be used in pharmaceutical compositions that include one or more other active pharmaceutical ingredients. The compounds may also be used in pharmaceutical compositions ATTORNEY DOCKET NO.: D0963.70010WO00

[0428] in which the compound disclosed herein or a pharmaceutically acceptable salt thereof is the only active ingredient.

[0429] Optical Isomers - Diastereomers - Geometric Isomers - Tautomers

[0430] Compounds disclosed herein may contain, e.g., one or more stereogenic centers and can occur as racemates, racemic mixtures, single enantiomers, individual diastereomers, and mixtures of diastereomers and / or enantiomers. The invention includes all such isomeric forms of the compounds disclosed herein. It is intended that all possible stereoisomers (e.g., enantiomers and / or diastereomers) in mixtures and as pure or partially purified compounds are included within the scope of this invention (i.e., all possible combinations of the stereogenic centers as pure compounds or in mixtures).

[0431] Some of the compounds described herein may contain bonds with hindered rotation such that two separate rotomers, or atropisomers, may be separated and found to have different biological activity which may be advantageous. It is intended that all of the possible atropisomers are included within the scope of this invention.

[0432] Some of the compounds described herein may contain olefinic double bonds, and unless specified otherwise, are meant to include both E and Z geometric isomers.

[0433] Some of the compounds described herein may exist with different points of attachment of hydrogen, referred to as tautomers. An example is a ketone and its enol form, known as keto-enol tautomers. The individual tautomers as well as mixtures thereof are encompassed by the invention.

[0434] Compounds disclosed herein having one or more asymmetric centers may be separated into diastereoisomers, enantiomers, and the like by methods well known in the art.

[0435] Alternatively, enantiomers and other compounds with chiral centers may be synthesized by stereospecific synthesis using optically pure starting materials and / or reagents of known configuration.

[0436] Metabolites - Prodrugs

[0437] The invention includes therapeutically active metabolites, where the metabolites themselves fall within the scope of the claims. The invention also includes prodrugs, which are compounds that are converted to the claimed compounds as they are being administered to a patient or after they have been administered to a patient. The claimed chemical structures of this application in some cases may themselves be prodrugs.

[0438] Isotopically Enriched Derivatives

[0439] The invention includes molecules which have been isotopically enriched at one or more position within the molecule. Thus, compounds enriched for deuterium fall within the scope of the claims. ATTORNEY DOCKET NO.: D0963.70010WO00

[0440] Methods of Preparing Myt1 inhibitors

[0441] Methods of preparing Myt1 inhibitors are known in the art (see, e.g., International Patent Publication No. WO 2022 / 213204, the entirety of which is incorporated herein by reference).

[0442] Combination Therapies

[0443] In some embodiments, the Myt1 inhibitor is administered as a combination therapy, i.e., in combination with a second active pharmaceutical ingredient. In some embodiments, the second active pharmaceutical ingredient is selected from a Wee1 inhibitor, a Fen1 inhibitor, a Topi inhibitor, an Rrm1 inhibitor, an Rrm2 inhibitor, an Aurkb inhibitor, a Top2A inhibitor, an ATR inhibitor, a Ttk inhibitor, an Sod1 inhibitor, an Sod2 inhibitor, a Bub1 inhibitor, a Cdc7 inhibitor, a Sae1 inhibitor, a Plk1 inhibitor, Uba2 inhibitor, a Dut inhibitor, an Hdac3 inhibitor, a Chekl inhibitor, an Aurka inhibitor, a Men1 inhibitor, a Doti I inhibitor, a Crebbp inhibitor, an Ezh2 inhibitor, a Plk4 inhibitor, an Haspin inhibitor, a Mettl3 inhibitor, a nucleoside analog, a platinum-based DNA damaging agent, or a combination thereof.

[0444] ATR Inhibitors

[0445] ATR inhibitors a compound that upon contacting the enzyme ATR kinase, whether in vitro, in cell culture, or in an animal, reduces the activity of ATR kinase, such that the measured ATR kinase IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain ATR inhibitors, the ATR kinase IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, the ATR kinase IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM).

[0446] Examples of ATR inhibitors are:

[0447] H2N

[0448]

[0449] elimusertib (BAY1895344) ceralasertib (AZD6738) berzosertib (VE-822) ATTORNEY DOCKET NO.: D0963.70010WO00

[0450] Gartisertib (VX-803, M4344) tuvusertib (M1774)

[0451]

[0452] SKLB-197

[0453] or a pharmaceutically acceptable salt thereof.

[0454] In one embodiment, an ATR inhibitor is a compound of formula (IV):

[0455] N R1

[0456]

[0457] (IV)

[0458] or a pharmaceutically acceptable salt thereof,

[0459] where

[0460] - is a double bond, and each Y is independently N or CR4; or - is a single bond, and each Y is independently NRY, carbonyl, or C(RY)2; where each RYis independently H or optionally substituted C1-6 alkyl;

[0461] R1is optionally substituted C1-6 alkyl or H;

[0462] R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or-Q-R5B;

[0463] R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl; each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0464] each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C1-9 heteroaryl, or-SO2R5A; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0465] each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted Ce-io aryl;

[0466] R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, –SO2R5A, or optionally substituted alkoxy;

[0467] each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted Ce-io aryl C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0468] Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted Ce-io arylene; and

[0469] X is hydrogen or halogen.

[0470] TheATR inhibitor may be, e.g., a compound of formula (V):

[0471]

[0472] (V);

[0473] or a pharmaceutically acceptable salt thereof,

[0474] where

[0475] each Y is independently N or CR4;

[0476] R1is optionally substituted C1-6 alkyl or H;

[0477] R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or-Q-R5B;

[0478] R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl;

[0479] each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl;

[0480] each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-io aryl C1-6 alkyl, optionally substituted Ce-io aryl, optionally substituted C1-9 heteroaryl, or-SO2R5A; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0481] each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted C6-10 aryl;

[0482] R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or optionally substituted alkoxy;

[0483] each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0484] Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted C6-10 arylene; and

[0485] X is hydrogen or halogen.

[0486] In some embodiments, in the compound of formula (IV) or (V):

[0487] each Y is independently N or CR4;

[0488] R1is H or optionally substituted C1-6 alkyl;

[0489] R2is optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, or-SO2R5A;

[0490] R3is optionally substituted C1-9 heteroaryl;

[0491] each R4is independently H or optionally substituted C1-6 alkyl;

[0492] each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, or-SO2R5A, where each R5Ais independently optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;

[0493] each R5Ais independently optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; and

[0494] each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-10 aryl C1-6 alkyl, optionally substituted C6-10 aryl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl.

[0495] Methods of making compounds of formula (IV) are described, e.g., in International Application No. PCT / US2019 / 051539, hereby incorporated by reference.

[0496] TheATR inhibitor may be, e.g., a compound of formula (IV-a):

[0497]

[0498] Y=Y ATTORNEY DOCKET NO.: D0963.70010WO00

[0499] (IV-a);

[0500] or a pharmaceutically acceptable salt thereof, where Y, R1, R2, R3, and R4are as described for formula (III).

[0501] TheATR inhibitor may be, e.g., a compound of formula (IV-b):

[0502]

[0503] (IV-b);

[0504] or a pharmaceutically acceptable salt thereof, where Y, R1, R2, R3, and R4are as described for formula (IV).

[0505] TheATR inhibitor may be, e.g., a compound of formula (IVA):

[0506]

[0507] (IVA);

[0508] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0509] TheATR inhibitor may be, e.g., a compound of formula (IVA-a):

[0510]

[0511] (IVA-a);

[0512] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0513] TheATR inhibitor may be, e.g., a compound of Formula (IVB): ATTORNEY DOCKET NO.: D0963.70010WO00

[0514] Q ^N^R1

[0515] R2AY '-N-R5

[0516] / =N

[0517]

[0518] R4

[0519] (IVB);

[0520] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0521] TheATR inhibitor may be, e.g., a compound of formula (IVB-a):

[0522] C ^°N1^*R1

[0523] R2AY '-N-R5

[0524] / =N

[0525]

[0526] R4

[0527] (IVB-a);

[0528] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0529] The ATR inhibitor may be, e.g., a compound of Formula (I I IC):

[0530] Q ^N^R1

[0531] R2f'Ys'N-R2'

[0532] N=Z

[0533]

[0534] R4

[0535] (I VC);

[0536] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0537] TheATR inhibitor may be, e.g., a compound of formula (IllC-a):

[0538] C ^°N1^*R1

[0539] R’ rYS ~'N-R’’

[0540] N=Z

[0541]

[0542] R4

[0543] (IVC-a); ATTORNEY DOCKET NO.: D0963.70010WO00

[0544] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0545] TheATR inhibitor may be, e.g., a compound of formula (IVD):

[0546] ^N^R1

[0547] R’ ~f 'N-R'

[0548]

[0549] ° R4

[0550] (IVD);

[0551] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0552] TheATR inhibitor may be, e.g., a compound of formula (IVD-a):

[0553] Q ^N^*R1

[0554] R2rYS 'N-R3

[0555]

[0556] ° R4

[0557] (IVD-a);

[0558] or a pharmaceutically acceptable salt thereof, where R1, R2, R3, and R4are as described for formula (IV).

[0559] Preferably, R1is methyl.

[0560] In some embodiments, R2may be, e.g., optionally substituted C3-8 cycloalkyl. For example, R2may be a group of formula (A):

[0561] « / wv

[0562] - R7

[0563] )n

[0564] (A)

[0565] where

[0566] n is 0, 1, 2, or 3; and

[0567] R7is hydrogen, alkylsulfonyl, cyano, -CON(RA)2, -SON(RA)2, optionally substituted C1-9 heteroaryl, hydroxy, or alkoxy, where each RAis independently H or alkyl; or both RA, together with the atom to which they are attached, combine to form C2-9 heterocyclyl.

[0568] In some embodiments, R2may be, e.g., optionally substituted C1-6 alkyl (e.g., optionally substituted tertiary C3-6 alkyl. For example, R2may be a group of formula (B):

[0569]

[0570] R7

[0571] (B) ATTORNEY DOCKET NO.: D0963.70010WO00

[0572] where R7is hydrogen, alkylsulfonyl, cyano, -CON(RA)2, -SON(RA)2, optionally substituted C1-9 heteroaryl, hydroxy, or alkoxy, where each RAis independently H or alkyl; or both RA, together with the atom to which they are attached, combine to form C2-9 heterocyclyl.

[0573] In some embodiments, R2may be, e.g., optionally substituted non-aromatic C2-9 heterocyclyl. In some embodiments, R2may be, e.g.:

[0574]

[0575] ATTORNEY DOCKET NO.: D0963.70010WO00

[0576]

[0577] ATTORNEY DOCKET NO.: D0963.70010WO00

[0578]

[0579] In some embodiments, R3may be, e.g., optionally substituted, monocyclic C1-9 heteroaryl including at least one nitrogen atom (e.g., two nitrogen atoms). For example, R3may be a group of formula (C):

[0580]

[0581] (C)

[0582] where A is optionally substituted, monocyclic C1-9 heteroaryl ring.

[0583] In some embodiments, A may be, e.g., a group of formula (C1):

[0584]

[0585] R8

[0586] (C1)

[0587] where R8is hydrogen, halogen, or optionally substituted C1-6 alkyl.

[0588] In some embodiments, R3may be, e.g.:

[0589]

[0590] In some embodiments, R3may be, e.g.:

[0591]

[0592] In some embodiments, R4may be, e.g., hydrogen. ATTORNEY DOCKET NO.: D0963.70010WO00

[0593] The ATR inhibitor may be, e.g., a compound listed in Table 2 or a pharmaceutically acceptable salt thereof.

[0594] Table 2

[0595]

[0596] ATTORNEY DOCKET NO.: D0963.70010WO00

[0597] MN"^ (HN"^

[0598] N^sZ "'- [<^O N^=Z ''"•■|<^O N-_zN., Nx / J xN. JM. J

[0599] CCJ N. \ ||

[0600] A22 T A23 A24

[0601] crN^ H2N^ X) I I

[0602] NHN^ HN^X nN"^

[0603] ,'|xy^x'O N^ / ''i^r^o N^ xN^ xN^^J

[0604] m v CXJ

[0605] A25I A26 A27 -OCT °^'N> |— — =N o— '

[0606] HN^\HN^

[0607] N^ / '‘|^w ^O N^ / |^^° A / N-K / N. J N-^N^XN^A N- _ xN\ / N. J <x y / ll P A28 A29 vA A30 Y v.^yx! J X11 / — v —sN

[0608] VSn

[0609] HN^HN^\HN^\

[0610] NiV N<= / '''■•[<^O

[0611] )*l^x>N'>x'N^xJ

[0612] / if i «NCLJA31vxy * A32 A33

[0613] S^N

[0614] ^=N HN^\

[0615] Ns=Z O

[0616] N^ xNk xN^xJ

[0617] a a vOr °'V AN

[0618] A34 TA43XAJLA y / ~N\ / A42Jl 1HN'NHN= / l'Z\kN-\> f j U 'N^

[0619] C°1

[0620] a a

[0621] A45 f| N A47 r<^N

[0622] A44°> JC^A / N~NHN-NH z V \ |^ -A^ O' xJ-L xA / / ~Nh

[0623] 0V 0 ^N 1 ^=N

[0624] C°1 a c° Nx

[0625] N. (i ^N

[0626] A48i i llHNA49 l| 7 N-NH

[0627] ~NA50°.. A xA,N~NH N-A X^ ^-N 0-J °

[0628]

[0629] ATTORNEY DOCKET NO.: D0963.70010WO00

[0630]

[0631] ATTORNEY DOCKET NO.: D0963.70010WO00

[0632]

[0633] ATTORNEY DOCKET NO.: D0963.70010WO00

[0634]

[0635] ATTORNEY DOCKET NO.: D0963.70010WO00

[0636] > D C

[0637] 00

[0638] a

[0639] A99 AN

[0640] °H|I JI N-NH / \® T N"^\^)T |a

[0641] QO" r

[0642] a N

[0643] A100rAN^ Zz

[0644] OHH I N-i VNH IA101?jXa. / *~nh\ / ^=N

[0645] H7 — '

[0646] —o

[0647] a N a N ^*

[0648] f| N A102 OH]J l N-NH

[0649] _ |X\^X / / \A103LAA / / ~NH / \ \ N^Xs^

[0650] \ / ^=N \ / ^=N

[0651] H / — 'H / - '

[0652] o—'

[0653] / W

[0654] N

[0655] I

[0656] a N ^

[0657] K^^N A104 / -^XA N-NH A105 °H|| I N-NH \ w / 'OHT,NX^ / —

[0658] ( ) ^=N

[0659] FF

[0660] O"1

[0661] a N

[0662] a

[0663] A106 90 / / 'T A107 AN

[0664] / \ JI J N-NH °\ Z<OH\ N~\^ ■r "

[0665] N

[0666] a N a

[0667] A108P1 A109 A

[0668] r if N hNf\ a / n~nhI > — A~NH^=N -N— T1V —VAHL ^N

[0669]

[0670] ATTORNEY DOCKET NO.: D0963.70010WO00

[0671] a

[0672] A110?!x Jk A111 r<^N

[0673] / — x 'y,Nx^? X Jk / / ~NH\

[0674] 0 □OUn

[0675] a a

[0676] r ^N jz‘z^N A112 OH|| 7 / / ~NHA113 °H|I 7 N-NH / — ^ y ^N^ xs^ / — N^X^0

[0677] ?y-N

[0678] c°x

[0679] N a

[0680] (i N

[0681] A114 QH|I J 7 / ~NHA11522H OHI] J1N-NH / \ \HN] |>^< X JI \ \ » / 2HV \? H\

[0682] Fx 2H A / 2H

[0683] ZNFFH2H2H2H

[0684] a

[0685] a

[0686] A116 XA1 1 7HO JO,N~NH l< S \ N XS^ I

[0687] c°x c°

[0688] N Nx A118 1 A119 I AN

[0689] IlN / — / N~NH\ / — \JxA £*~NH

[0690] / N\_J<O^ZN' X^

[0691] a

[0692] C Nx ^*

[0693] A120 (C*N

[0694] / — \ ll N-NHA121N FN\ / ^OH\,N" X^ Yrr'H T A *=N \c^ / ^=N

[0695] H

[0696]

[0697] ATTORNEY DOCKET NO.: D0963.70010WO00

[0698] >

[0699] 00

[0700] 0 L A JJ*)

[0701] a

[0702] A122 OH|| 7

[0703] jM

[0704] y-NHA123yN~NH7 - N'A ( _ y *=N I

[0705] zz-N

[0706] J! b z H

[0707] z

[0708] HN--\

[0709] N-a / ''YAO b o?

[0710] Q <0 z 7= — N J< J A < 7 A125 J

[0711] O M —

[0712] T

[0713] A>>^NH2

[0714] HN

[0715] *|^ ‘ 0 ^^. Nxy-NxA A127NObJ

[0716] T0O \—

[0717] si / " NH2. On tX', (g / X0 -Z>=N ), Z W y— z- \ _ \ _ > O \ _ \ _' c O'''

[0718] HN b A ■Hl^ A

[0719] b A ■

[0720] & ' Jw 0 NA "''-[lAo YV, / Z / \ H

[0721] X / / \ Zz z- N^Nxy-N«A

[0722] Z Z- A128Nbk J A129 AJL JJ J I HO

[0723] 0[0P fj rrNH2

[0724] ^■No c

[0725] 5I c CX ™ A. < < N^Z 'j'w 0

[0726] NCL J

[0727] A1310

[0728] CX / NHzFF

[0729] A A N

[0730] fl I1IlNA132 no JQ P A133H1AA A U.N" VN / A u - N'NON HF \ ]JNH FA-°

[0731] F

[0732]

[0733] ATTORNEY DOCKET NO.: D0963.70010WO00

[0734] a (X

[0735] N N

[0736] A134o A A135

[0737] K JL X, N-NH

[0738] O-X '•=N AvC 'NV >< 7NHNz0-^

[0739] c° Nx a A136 Q

[0740] II 7 N-NHA137XA,~

[0741] N XN

[0742] xNH4 " A NXX

[0743] °AUn' I if

[0744] CF =

[0745] (S

[0746] Y MH,.

[0747] A138 X A139 / X-! f 4 1 1 X, »

[0748] 4 H YT CX” X AM X.... NI 1 X-NH Ck...-s.

[0749] 11 1

[0750] I Js!*;

[0751] N

[0752] I "NHS

[0753] A140t f Y's A141 '1 IN

[0754] I 1 AN’; H r" Y

[0755] [ Ai J' Y AV / <>

[0756] ,. Q%p r

[0757] A 142 p.4... r A143 p — § — p Y^'N

[0758] j |! J r-. H 1 X YYW f V"

[0759] U ■"

[0760] 0.

[0761] X

[0762] F i O 'X

[0763] A144 4NA145 Ag.o A

[0764] j. Jy I j..1

[0765] :;: N

[0766]

[0767] ATTORNEY DOCKET NO.: D0963.70010WO00

[0768]

[0769] Compound A121 may be referred to interchangeably using the name “camonsertib” herein. An ATR inhibitor may be isotopically enriched (e.g., enriched for deuterium).

[0770] Further non-limiting examples of ATR inhibitors include, e.g., those described in, e.g., International Application Publication Nos. WO 2020087170, WO 2018218197, WO 2020259601, WO 2019036641, WO 2020049017, WO 2019154365, WO 2020103897, WO 2021233376, WO 2022028598, WO 2022012484, WO 2022002245, and WO 2022002243, each of which is incorporated by reference herein; U. S. Patent Nos. 11,028,076, 10,745,420, 10,301,324, 10,196,405, 9,663,535, 9,549,932, 8,552,004, and 8,841,308, each of which is incorporated by reference herein; and U. S. Patent Application Publication Nos. 2019 / 0055240 and 2019 / 0300547, each of which is incorporated by reference herein.

[0771] An ATR inhibitor may be isotopically enriched (e.g., enriched for deuterium). ATTORNEY DOCKET NO.: D0963.70010WO00

[0772] Aurka Inhibitors

[0773] Aurka inhibitors may be compounds that upon contacting Aurka, whether in vitro, in cell culture, or in an animal, reduce the activity of Aurka, such that the measured Aurka IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Aurka inhibitors, Aurka IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Aurka IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Aurka inhibitors are: MK0547, barasertib (AZD1152), PHA739358, AT9283, AMG900, SNS-314, TAK-901, CYC116, GSK1070916, PF03814735, and pharmaceutically acceptable salts thereof. Exemplary Aurka inhibitors are also disclosed in US 6,977,259; US 6,919,338; US 7,105,669; US 7,214,518; US 7,235,559; US 7,402,585; US 7,709,479; US 8,026,246; US 8,138,338; US 8,377,983; US 9,567,329; US 9,637,474; US 20060178382; US 20090029992; and US 20190352297; the Aurka inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0774] Aurkb Inhibitors

[0775] Aurkb inhibitors may be compounds that upon contacting Aurkb, whether in vitro, in cell culture, or in an animal, reduce the activity of Aurkb, such that the measured Aurkb IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Aurkb inhibitors, Aurkb IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Aurkb IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Aurkb inhibitors are: MLN8237, MK0547, MLN8054, PHA739358, AT9283, AMG900, MK5108, SNS314, TAK901, CYC116, ENMD2076, and pharmaceutically acceptable salts thereof. Exemplary Aurkb inhibitors are also disclosed in US 7,560,551; US 7,977,477; US 8,110,573; and US 20110293745; the Aurkb inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0776] Bub1 Inhibitors

[0777] Bub1 inhibitors may be compounds that upon contacting Bub1, whether in vitro, in cell culture, or in an animal, reduce the activity of Bub1, such that the measured Bub1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Bub1 inhibitors, Bub1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Bub1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Bub1 inhibitors are: BAY-320, BAY-419, BAY1816032 and pharmaceutically acceptable salts thereof. Exemplary Bub1 inhibitors are also disclosed in US 9,265,763; US 9,416,125; US 9,745,285; US 10,266,548; US 10,428,044; US 20150141372; US 20160145267; US 20160046604; US 20160046610; US 20170275269; US 20170305882; the Bub1 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0778] Cdc7 Inhibitors

[0779] Cdc7 inhibitors may be compounds that upon contacting Cdc7, whether in vitro, in cell culture, or in an animal, reduce the activity of Cdc7, such that the measured Cdc7 IC50 is 10 pM or less (e.g., ATTORNEY DOCKET NO.: D0963.70010WO00

[0780] 5 pM or less or 1 pM or less). For certain Cdc7 inhibitors, Cdc7 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Cdc7 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Cdc7 inhibitors are: SRA141, TAK931, and pharmaceutically acceptable salts thereof. Exemplary Cdc7 inhibitors are also disclosed in US 7,279,575; US 8,314,121; US 8,383,624; US 8,658,662; US 8,691,828; US 9,156,824; US 9,180,105; US 9,974,795; US 10,745,510; US 20050043346; US 20050256121; US 20070293491; US 20190336502; and US 20200093828; the Cdc7 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0781] Chekl Inhibitors

[0782] Chekl inhibitors may be compounds that upon contacting Chekl, whether in vitro, in cell culture, or in an animal, reduce the activity of Chekl, such that the measured Chekl IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Chekl inhibitors, Chekl IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Chekl IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Chekl inhibitors are: SRA737 and pharmaceutically acceptable salts thereof. Exemplary Chekl inhibitors are also disclosed in US 7,067,506; US 8,093,244; US 8,410,279; US 8,530,468; US 8,618,121; US 8,916,591; US 9,067,920; US 9,440,976; US 10,189,818; US 10,822,327; US 20090182001; US 20090233896; US 20090258852; US20090270416; US 20090275570; US 20150368244; US 20180369202; and US 20200397796; the Chekl inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0783] Crebbp Inhibitors

[0784] Crebbp inhibitors may be compounds that upon contacting Crebbp, whether in vitro, in cell culture, or in an animal, reduce the activity of Crebbp, such that the measured Crebbp IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Crebbp inhibitors, Crebbp IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Crebbp IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Crebbp inhibitors are: CPI4, CCS1477, E7386, NEO1132, NEO2734, PRI724, C82, BC001, C646, EML425, CBP30, and pharmaceutically acceptable salts thereof. Exemplary Crebbp inhibitors are also disclosed in US 9,763,922; US 10,206,931; US 10,696,655; US 10,870,648; US 20190270797; US 20190298729; and US 20190308978; the Crebbp inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0785] Dotll Inhibitors

[0786] Doti I inhibitors may be compounds that upon contacting Dotll, whether in vitro, in cell culture, or in an animal, reduce the activity of Dotll, such that the measured Dotll IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Dotll inhibitors, Doti I IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Dotll IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Doti I inhibitors ATTORNEY DOCKET NO.: D0963.70010WO00

[0787] are: pinometostat (EPZ5676) and pharmaceutically acceptable salts thereof. Exemplary Doti I inhibitors are also disclosed in US 8,722,877; US 9,458,165; US 10,112,968; US 20140100184; US 20150342979; and US 20170335402; the Doti I inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0788] Dut Inhibitors

[0789] Dut inhibitors may be compounds that upon contacting Dut, whether in vitro, in cell culture, or in an animal, reduce the activity of Dut, such that the measured Dut IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Dut inhibitors, Dut IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Dut IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Dut inhibitors are: TAS114 and pharmaceutically acceptable salts thereof. Exemplary Dut inhibitors are also disclosed in US 7,601,702; US 8,530,490; US 9,790,214; US 9,809,571; US 10,544,105; US 10,562,860; US 10,570,100; US 10,577,321; US 10,829,457; US 10,858,344; US 20110212467; US 20190270756; US 20190330158; US 20190330210; and US 20200039966; the Dut inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0790] Ezh2 Inhibitors

[0791] Ezh2 inhibitors may be compounds that upon contacting Ezh2, whether in vitro, in cell culture, or in an animal, reduce the activity of Ezh2, such that the measured Ezh2 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Ezh2 inhibitors, Ezh2 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Ezh2 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Ezh2 inhibitors are: EPZ-6438, GSK126, and pharmaceutically acceptable salts thereof. Exemplary Ezh2 inhibitors are also disclosed in US 8,691,507; US 9,394,283; US 9,889,138; US 10,166,238; US 10,040,782; US 10,457,640; US 10,633,371; US 10,647,700; US 10,786,511; US 20190328743; US 20190345139; and US 20210052595; the Ezh2 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0792] Haspin Inhibitors

[0793] Haspin inhibitors may be compounds that upon contacting Haspin, whether in vitro, in cell culture, or in an animal, reduce the activity of Haspin, such that the measured Haspin IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Haspin inhibitors, Haspin IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Haspin IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Haspin inhibitors are: SEL120 and pharmaceutically acceptable salts thereof. Exemplary Haspin inhibitors are also disclosed in US 20130102627 and US 20130231360; the Haspin inhibitors disclosed therein are incorporated herein by reference in their entirety. ATTORNEY DOCKET NO.: D0963.70010WO00

[0794] Hdac3 Inhibitors

[0795] Hdac3 inhibitors may be compounds that upon contacting Hdac3, whether in vitro, in cell culture, or in an animal, reduce the activity of Hdac3, such that the measured Hdac3 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Hdac3 inhibitors, Hdac3 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Hdac3 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Hdac3 inhibitors are: RGFP966 and pharmaceutically acceptable salts thereof. Exemplary Hdac3 inhibitors are also disclosed in US 8,716,344; US 9,096,549; US 10,029,988; US 10,059,723; and US 20190216754; the Hdac3 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0796] Fen1 Inhibitors

[0797] Fen1 inhibitors may be compounds that upon contacting Fen1, whether in vitro, in cell culture, or in an animal, reduce the activity of Fen1, such that the measured Fen1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Fen1 inhibitors, Fen1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Fen1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Fen1 inhibitors are: C8 (PMID: 32719125), SC13, Fen1-IN-3, and pharmaceutically acceptable salts thereof. Exemplary Fen1 inhibitors are also disclosed in US 20200237763 and US 7,927,790; the Fen1 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0798] Men1 Inhibitors

[0799] Men1 inhibitors may be compounds that upon contacting Men1, whether in vitro, in cell culture, or in an animal, reduce the activity of Men1, such that the measured Men1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Men1 inhibitors, Men1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Men1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Men1 inhibitors are: MI3454, SNDX5613, VTP50469, KO539, and pharmaceutically acceptable salts thereof. Exemplary Men1 inhibitors are also disclosed in US 8,242,078’ US 9,212,180; US 10,077,271; US 10,526,341; US 10,611,778; US 10,745,409; US 10,752,639; US 10,781,218; US 10,899,738; US 20170119769; US 20190010167; US 20190211036; US 20200022953; US 20200216471; and US 20200223853; the Men1 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0800] Mettl3 Inhibitors

[0801] Mettl3 inhibitors may be compounds that upon contacting Mettl3, whether in vitro, in cell culture, or in an animal, reduce the activity of Mettl3, such that the measured Mettl3 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Mettl3 inhibitors, Mettl3 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Mettl3 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of ATTORNEY DOCKET NO.: D0963.70010WO00

[0802] Mettl3 inhibitors are: UZH1a, sTC-15, and pharmaceutically acceptable salts thereof. Exemplary Mettl3 inhibitors are also disclosed in US 20160264934 and WO 2020201773; the Mettl3 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0803] Nucleoside Analogs

[0804] Nucleoside analogs may be compounds that can act as antimetabolites by interfering with nucleotide production, or by acting as chain terminators in DNA lengthening by polymerase enzymes, either in cell culture, or in an animal. For certain nucleoside analogs, biological activity make occur at 10 pM or less (e.g., 5 pM or less or 1 pM or less) and could be as low as 100 pM or 10 pM.

[0805] Preferably, nucleoside analog activity will occur at 1 nM to 1 pM (e.g., 1 nM to 750 nM, 1 nM to 500 nM, or 1 nM to 250 nM). Examples of nucleoside analogs are cytarabine, gemcitabine, mercaptopurine, azacytidine, cladribine, decitabine, fluorouracil, floxuridine, fludarabine or nelarabine.

[0806] Plk1 Inhibitors

[0807] Plk1 inhibitors may be compounds that upon contacting Plk1, whether in vitro, in cell culture, or in an animal, reduce the activity of Plk1, such that the measured Plk1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Plk1 inhibitors, Plk1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Plk1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Plk1 inhibitors are: BI2536, BI6727, TAK960, NMSP937, GSK461364, and pharmaceutically acceptable salts thereof. Exemplary Plk1 inhibitors are also disclosed in US 7,504,513; US 7,517,873; US 7,851,495; US 7,977,336; US 8,101,628; US 8,129,387; US 8,278,299; US 9,175,038; US 9,175,357; US 20070185133; US 20080015192; US 20100278833; US 20150368209; US 20170283445; and US 20200247796; the Plk1 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0808] Plk4 Inhibitors

[0809] Plk4 inhibitors may be compounds that upon contacting Plk4, whether in vitro, in cell culture, or in an animal, reduce the activity of Plk4, such that the measured Plk4 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Plk4 inhibitors, Plk4 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Plk4 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM).

[0810] Exemplary Plk4 inhibitors include compounds of formula (VI):

[0811]

[0812] ATTORNEY DOCKET NO.: D0963.70010WO00

[0813] or a pharmaceutically acceptable salt thereof,

[0814] wherein

[0815] n is 0, 1, 2, 3, or 4;

[0816] m is 0, 1, or 2;

[0817] L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted C6-10 aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;

[0818] R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;

[0819] R1bis hydrogen; or

[0820] R1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;

[0821] A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce-io aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl;

[0822] each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce-ioaryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl Ci-e alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;

[0823] R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor -N(R3B)2

[0824] each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl; ATTORNEY DOCKET NO.: D0963.70010WO00

[0825] X is N, and R4is absent; orX is C, and R4is hydrogen, halogen, cyano, optionally substituted amino, optionally substituted acyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;

[0826] R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;

[0827] Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted Ce-io arylene, or optionally substituted C3-8 cycloalkylene;

[0828] R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce-ioaryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;

[0829] R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; and R6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

[0830] The Plk4 inhibitor may be, e.g., a compound of formula (VI I):

[0831]

[0832] or a pharmaceutically acceptable salt thereof; wherein n, A, L, R1a, R1b, R2A, R2B, R3, R5, and R6are as defined in formula (VI).

[0833] The Plk4 inhibitor may be, e.g., a compound of formula (VIII):

[0834]

[0835] (VIII)

[0836] or a pharmaceutically acceptable salt thereof; wherein n, A, L, R1a, R1b, R2A, R2B, R3, R4, R5, and R6are as defined in formula (VI). ATTORNEY DOCKET NO.: D0963.70010WO00

[0837] The Plk4 inhibitor may be, e.g., a compound of formula (Vll-A):

[0838]

[0839] or a pharmaceutically acceptable salt thereof; wherein n, L, R1a, R1b, R2B, R3, R5, and R6are as defined in formula (VI).

[0840] The Plk4 inhibitor may be, e.g., a compound of formula (Vlll-A):

[0841]

[0842] (Vlll-A)

[0843] or a pharmaceutically acceptable salt thereof; wherein n, L, R1a, R1b, R2B, R3, R4, R5, and R6are as defined in formula (VI).

[0844] The Plk4 inhibitor may be, e.g., a compound of formula (Vll-B):

[0845]

[0846] or a pharmaceutically acceptable salt thereof; wherein n, L, R1a, R1b, R2A, R2B, R3, R5, and R6are as defined in formula (VI).

[0847] The Plk4 inhibitor may be, e.g., a compound of formula (Vlll-B):

[0848]

[0849] or a pharmaceutically acceptable salt thereof; wherein n, L, R1a, R1b, R2a, R2B, R3, R4, R5, and R6are as defined in formula (VI).

[0850] The Plk4 inhibitor may be, e.g., a compound of formula (Vll-C): ATTORNEY DOCKET NO.: D0963.70010WO00

[0851]

[0852] (Vll-C)

[0853] or a pharmaceutically acceptable salt thereof wherein n, L, R1a, R1b, R3, R5, and R6are as defined in formula (VI).

[0854] The Plk4 inhibitor may be, e.g., a compound of formula (Vlll-C):

[0855] R6

[0856]

[0857] (Vlll-C)

[0858] or a pharmaceutically acceptable salt thereof wherein n, L, R1a, R1b, R3, R4, R5, and R6are as defined in formula (VI).

[0859] The Plk4 inhibitor may be, e.g., a compound of formula (Vll-D):

[0860]

[0861] or a pharmaceutically acceptable salt thereof wherein n, L, R1a, R1b, R3, R5, and R6are as defined in formula (VI).

[0862] The Plk4 inhibitor may be, e.g., a compound of formula (Vlll-D):

[0863] R6

[0864]

[0865] (Vlll-D)

[0866] or a pharmaceutically acceptable salt thereof wherein n, L, R1a, R1b, R3, R4, R5, and R6are as defined in formula (VI).

[0867] The Plk4 inhibitor may be, e.g., a compound listed in Table 3 or a pharmaceutically acceptable salt thereof. ATTORNEY DOCKET NO.: D0963.70010WO00

[0868] Table 3

[0869]

[0870] ATTORNEY DOCKET NO.: D0963.70010WO00

[0871]

[0872] ATTORNEY DOCKET NO.: D0963.70010WO00

[0873]

[0874] ATTORNEY DOCKET NO.: D0963.70010WO00

[0875]

[0876] ATTORNEY DOCKET NO.: D0963.70010WO00

[0877]

[0878] ATTORNEY DOCKET NO.: D0963.70010WO00

[0879]

[0880] ATTORNEY DOCKET NO.: D0963.70010WO00

[0881]

[0882] ATTORNEY DOCKET NO.: D0963.70010WO00

[0883]

[0884] ATTORNEY DOCKET NO.: D0963.70010WO00

[0885]

[0886] ATTORNEY DOCKET NO.: D0963.70010WO00

[0887]

[0888] ATTORNEY DOCKET NO.: D0963.70010WO00

[0889]

[0890] ATTORNEY DOCKET NO.: D0963.70010WO00

[0891]

[0892] ATTORNEY DOCKET NO.: D0963.70010WO00

[0893]

[0894] ATTORNEY DOCKET NO.: D0963.70010WO00

[0895]

[0896] ATTORNEY DOCKET NO.: D0963.70010WO00

[0897]

[0898] ATTORNEY DOCKET NO.: D0963.70010WO00

[0899]

[0900] ATTORNEY DOCKET NO.: D0963.70010WO00

[0901]

[0902] ATTORNEY DOCKET NO.: D0963.70010WO00

[0903]

[0904] ATTORNEY DOCKET NO.: D0963.70010WO00

[0905]

[0906] ATTORNEY DOCKET NO.: D0963.70010WO00

[0907]

[0908] ATTORNEY DOCKET NO.: D0963.70010WO00

[0909]

[0910] ATTORNEY DOCKET NO.: D0963.70010WO00

[0911]

[0912] ATTORNEY DOCKET NO.: D0963.70010WO00

[0913]

[0914] ATTORNEY DOCKET NO.: D0963.70010WO00

[0915]

[0916] ATTORNEY DOCKET NO.: D0963.70010WO00

[0917]

[0918] ATTORNEY DOCKET NO.: D0963.70010WO00

[0919]

[0920] ATTORNEY DOCKET NO.: D0963.70010WO00

[0921]

[0922] ATTORNEY DOCKET NO.: D0963.70010WO00

[0923]

[0924] ATTORNEY DOCKET NO.: D0963.70010WO00

[0925]

[0926] ATTORNEY DOCKET NO.: D0963.70010WO00

[0927]

[0928] ATTORNEY DOCKET NO.: D0963.70010WO00

[0929]

[0930] Further, non-limiting examples of Plk4 inhibitors include centrinone, CFI-400945, and pharmaceutically acceptable salts thereof. Additional non-limiting examples of Plk4 inhibitors are also disclosed in US 10,752,612; US 20190070190; US 20200383990; and WO 2023159307; the Plk4 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0931] Rrm1 and Rrm2 Inhibitors

[0932] Rrm1 inhibitors may be compounds that upon contacting Rrm1, whether in vitro, in cell culture, or in an animal, reduces the activity of Rrm1, such that the measured Rrm1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Rrm1 inhibitors, Rrm1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Rrm1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM).

[0933] Rrm2 inhibitors may be compounds that upon contacting Rrm2, whether in vitro, in cell culture, or in an animal, reduce the activity of Rrm2, such that the measured Rrm2 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Rrm2 inhibitors, Rrm2 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Rrm2 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Rrm2 inhibitors are: motexafin gadolinium, hydroxyurea, fludarabine, cladribine, tezacitabine, triapine, and pharmaceutically acceptable salts thereof. Exemplary Rrm2 inhibitors are also disclosed in US 4,188,378; US 4,357,324; and US 2019 / 0161461; the Rrm2 inhibitors disclosed therein are incorporated herein by reference in their entirety. ATTORNEY DOCKET NO.: D0963.70010WO00

[0934] Sae1 Inhibitors

[0935] Sae1 inhibitors may be compounds that upon contacting Sae1, whether in vitro, in cell culture, or in an animal, reduce the activity of Sae1, such that the measured Sae1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Sae1 inhibitors, Sae1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Sae1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Sae1 inhibitors are: ML792 and pharmaceutically acceptable salts thereof. Exemplary Sae1 inhibitors are also disclosed in US 7,951,810; US 8,008,307; US 8,207,177; US 9,683,003; and US 9,695,154; the Sae1 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0936] Sod1 Inhibitors

[0937] Sod1 inhibitors may be compounds that upon contacting Sod1, whether in vitro, in cell culture, or in an animal, reduce the activity of Sod1, such that the measured Sod1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Sod1 inhibitors, Sod1 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Sod1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Sod1 inhibitors are: LCS1, ATN-224, pyrimethamine, and pharmaceutically acceptable salts thereof.

[0938] Sod2 Inhibitors

[0939] Sod2 inhibitors may be compounds that upon contacting Sod2, whether in vitro, in cell culture, or in an animal, reduce the activity of Sod2, such that the measured Sod2 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Sod2 inhibitors, Sod2 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Sod2 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Sod2 inhibitors are: LCS1, ATN-224, pyrimethamine, and pharmaceutically acceptable salts thereof.

[0940] Topi Inhibitors

[0941] Topi inhibitors may be compounds that upon contacting Topi, whether in vitro, in cell culture, or in an animal, reduce the activity of Topi, such that the measured Topi IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Topi inhibitors, Topi IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Topi IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Topi inhibitors are: irinotecan (including its metabolite irino metab), topotecan, camptothecin, lamellarin D, and pharmaceutically acceptable salts thereof. Exemplary Topi inhibitors are also disclosed in US 4,604,463; US 4,894,456; and US 5,004,758; the Topi inhibitors disclosed therein are incorporated herein by reference in their entirety. ATTORNEY DOCKET NO.: D0963.70010WO00

[0942] Top2 Inhibitors

[0943] Top2 inhibitors may be compounds that upon contacting Top2, whether in vitro, in cell culture, or in an animal, reduce the activity of Top2, such that the measured Top2 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Top2 inhibitors, Top2 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Top2 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Top2 inhibitors are: etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, and pharmaceutically acceptable salts thereof. Exemplary Top2 inhibitors are also disclosed in US 3,590,028; US 3,933,827; US 3,989,598; US 4,258,191; US 4,464,529; and US 4,965,348; the Top2 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0944] Ttk Inhibitors

[0945] Ttk inhibitors may be compounds that upon contacting Ttk, whether in vitro, in cell culture, or in an animal, reduce the activity of Ttk, such that the measured Ttk IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Ttk inhibitors, Ttk IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Ttk IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Ttk inhibitors are:

[0946] BAY1217389 and pharmaceutically acceptable salts thereof. Exemplary Ttk inhibitors are also disclosed in US 8,551,980; US 8,729,082; US 9,199,999; US 9,212,184; US 9,284,317; US 9,340,528; US 9,388,140; US 9,388,177; US 9,468,642; US 9,512,126; US 9,512,130; US 9,555,022; US 9,586,958; US 9,663,510; US 9,670,202; US 2017 / 0217946; US 2017 / 0305912; US 2017 / 0334899; US 2017 / 0342064; US 9,676,766; Wengner et al., Mol. Cancer Then, 15:583-592, 2016; Zaman et al., Mol. Cancer Then, 16:2609-2617, 2017; Mason et al., Proc. Nat’l Acad. Sci. U. S. A., 21:3127-3132, 2017; and Riggs et al., J. Med. Chem., 62:4401-4410, 2019; the Ttk inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0947] Uba2 Inhibitors

[0948] Uba2 inhibitors may be compounds that upon contacting Uba2, whether in vitro, in cell culture, or in an animal, reduce the activity of Uba2, such that the measured Uba2 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Uba2 inhibitors, Uba2 IC50 may be 100 nM or less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Uba2 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM). Examples of Uba2 inhibitors are: TAK981 and pharmaceutically acceptable salts thereof. Exemplary Uba2 inhibitors are also disclosed in US 9,045,483; the Uba2 inhibitors disclosed therein are incorporated herein by reference in their entirety.

[0949] Wee1 Inhibitors

[0950] Wee1 inhibitors may be compounds that upon contacting Wee1, whether in vitro, in cell culture, or in an animal, reduce the activity of Wee1, such that the measured Wee1 IC50 is 10 pM or less (e.g., 5 pM or less or 1 pM or less). For certain Wee 1 inhibitors, Wee1 IC50 may be 100 nM or ATTORNEY DOCKET NO.: D0963.70010WO00

[0951] less (e.g., 10 nM or less, or 1 nM or less) and could be as low as 100 pM or 10 pM. Preferably, Wee1 IC50 is 0.1 nM to 1 pM (e.g., 0.1 nM to 750 nM, 0.1 nM to 500 nM, or 0.1 nM to 250 nM).

[0952] Examples of Wee1 inhibitors are:

[0953] Adavosertib Azenosertib W02020192581

[0954] Example 59

[0955] WO 2019085933 Example WO2018171633 WO 2018090939

[0956] Example 4

[0957] WO 202256680 WO 2022155202 Example P11

[0958] Example 41

[0959]

[0960] and pharmaceutically acceptable salts thereof.

[0961] Further Examples of Wee1 inhibitors can be found in, e.g., US Patent Nos.: 8,710,065;

[0962] 8,716,297; 8,791,125, 9,181,239; 9,850,247; 11,332,473; 11,345,711; 11,208,413; 11,248,006;

[0963] 11,613,545; and 11,261,192; US Patent Application No. 17 / 441,085; and International Patent Publication Nos.: WO 2013012681; WO 2018090939; WO 2018171633; WO 2019028008; WO

[0964] 2019085933; WO 2019173082; WO 2020192581; WO 2020210320; WO 2020210377; WO 2021073491; WO 2021207598; WO 2022155202; WO 2022188802; WO 2022251224; and WO 2022256680; the Wee1 inhibitors of which are all incorporated herein by reference in their entirety.

[0965] Platinum-Based DNA-Damaging Agent

[0966] Platinum-based DNA-damaging agents are coordination compounds of Pt(ll) or Pt(IV), typically known in the art as platins. Platinum-based DNA-damaging agents include at least two coordination sites at the platinum center that are occupied by nitrogenous spectator ligand(s). The ATTORNEY DOCKET NO.: D0963.70010WO00

[0967] nitrogenous spectator ligands are monodentate or bidentate ligands, in which the donor atom is an sp3- orsp2-hybridized nitrogen atom within the ligand. Non-limiting examples of nitrogenous spectator ligands are ammonia, 1,2-cyclohexanediamine, a picoline, phenanthrin, or 1,6-hexanediamine. Nonlimiting examples of platinum-based DNA-damaging agents include cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, and satraplatin.

[0968] Methods Of Treatment

[0969] The methods disclosed herein may be used to treat a cancer (e.g., colorectal cancer, uterine cancer, bladder cancer, cervical cancer, upper gastrointestinal cancer, ovarian cancer, or breast cancer) in a subject, the method including administering to the subject in need thereof a therapeutically effective amount of a Myt1 inhibitor. The cancer may be identified as having a truncating biallelic loss-of-fu notion mutation described herein. Alternatively, the cancer may have been previously identified as having a truncating biallelic loss-of-function mutation described herein.

[0970] The methods disclosed herein may be used to induce cell death in a cancer cell, the method including administering to the cell a therapeutically effective amount of a Myt1 inhibitor. The cancer cell may be identified as having a truncating biallelic loss-of-function mutation described herein. Alternatively, the cancer cell may have been previously identified as having a truncating biallelic loss-of-function mutation described herein.

[0971] In some instances, the methods disclosed herein may include the step of administering to the subject in need thereof a therapeutically effective amount of a membrane-associated tyrosine and threonine-specific cdc2 inhibitory kinase (Myt1) inhibitor and a therapeutically effective amount of a second therapeutic. The second therapeutic may be, e.g., Wee1 inhibitor, Fen1 inhibitor, Topi inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Aurkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod1 inhibitor, Sod2 inhibitor, Bub1 inhibitor, Cdc7 inhibitor, Sae1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Dut inhibitor, Hdac3 inhibitor, Chekl inhibitor, Aurka inhibitor, Men1 inhibitor, Doti I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinum-based DNA damaging agent, or a combination thereof.

[0972] Administration of a Myt1 inhibitor to a subject with a cancer which has been previously identified as having a truncating biallelic loss-of-function mutation, or a cancer known to have a truncating biallelic loss-of-function mutation, may increase the susceptibility of the cancer to a replication stress inducing therapeutic (such as Myt1). This may result in premature mitosis of a cell, resulting in a daughter cell or daughter cells which may be unviable (e.g., due to induced DNA breaks or chromosome pulverization), ultimately resulting in the death of the cancer cell. In some embodiment, inducing the replication stress of the cancer cell may be increased by administering a second therapeutic (e.g., a Wee1 inhibitor, a Fen1 inhibitor, a Topi inhibitor, an Rrm1 inhibitor, an Rrm2 inhibitor, an Aurkb inhibitor, a Top2A inhibitor, an ATR inhibitor, a Ttk inhibitor, an Sod1 inhibitor, an Sod2 inhibitor, a Bub1 inhibitor, a Cdc7 inhibitor, an Sae1 inhibitor, a Plk1 inhibitor, a Uba2 inhibitor, a Dut inhibitor, an Hdac3 inhibitor, a Chekl inhibitor, an Aurka inhibitor, a Men1 inhibitor, a Doti I inhibitor, a Crebbp inhibitor, an Ezh2 inhibitor, a Plk4 inhibitor, an Haspin inhibitor, a Mettl3 inhibitor, a nucleoside analog, a platinum-based DNA damaging agent, or a combination thereof). ATTORNEY DOCKET NO.: D0963.70010WO00

[0973] Compounds disclosed herein may be administered by a route selected from the group consisting of oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intraorbital, intraventricular, intraspinal, intraperitoneal, intranasal, inhalation, intratumoral, and topical administration.

[0974] In some embodiments, the Myt1 inhibitor is administered before the second agent (e.g., within 1 week, within 6 days, within 5 days, within 4 days, within 3 days, within 2 days, within 1 day, or within 12 hours). In some embodiments, the Myt1 inhibitor is administered after the second agent (e.g., within 1 week, within 6 days, within 5 days, within 4 days, within 3 days, within 2 days, within 1 day, or within 12 hours). In some embodiments, the Myt1 inhibitor is co-administered with the second agent. In some embodiments, the Myt1 inhibitor is administered intermittently (e.g., 1 day / week, 2 days / week, or 3 days / week). In some embodiments, the second agent is administered on a continuous daily basis.

[0975] Pharmaceutical Compositions

[0976] The compounds used in the methods described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo. Pharmaceutical compositions typically include a compound as described herein and a pharmaceutically acceptable excipient. Certain pharmaceutical compositions may include one or more additional pharmaceutically active agents described herein.

[0977] The compounds described herein can also be used in the form of the free base, in the form of salts, zwitterions, solvates, or as prodrugs, or pharmaceutical compositions thereof. All forms are within the scope of the invention. The compounds, salts, zwitterions, solvates, prodrugs, or pharmaceutical compositions thereof, may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds used in the methods described herein may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration, and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0978] For human use, a compound of the invention can be administered alone or in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. Pharmaceutical compositions for use in accordance with the present invention thus can be formulated in a conventional manner using one or more physiologically acceptable carriers including excipients and auxiliaries that facilitate processing of a compound of the invention into preparations which can be used pharmaceutically.

[0979] This invention also includes methods of administering pharmaceutical compositions which can contain one or more pharmaceutically acceptable carriers. In making the pharmaceutical compositions of the invention, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or ATTORNEY DOCKET NO.: D0963.70010WO00

[0980] other container. When the excipient serves as a diluent, it can be a solid, semisolid, or liquid material (e.g., normal saline), which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, and soft and hard gelatin capsules. As is known in the art, the type of diluent can vary depending upon the intended route of administration. The resulting compositions can include additional agents, e.g., preservatives.

[0981] The excipient or carrier is selected on the basis of the mode and route of administration. Suitable pharmaceutical carriers, as well as pharmaceutical necessities for use in pharmaceutical formulations, are described in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), a well-known reference text in this field, and in the USP / NF (United States Pharmacopeia and the National Formulary). Examples of suitable excipients are lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include: lubricating agents, e.g., talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents, e.g., methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents. Other exemplary excipients are described in Handbook of Pharmaceutical Excipients, 6th Edition, Rowe et al., Eds., Pharmaceutical Press (2009).

[0982] These pharmaceutical compositions can be manufactured in a conventional manner, e.g., by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Methods well known in the art for making formulations are found, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), and Encyclopedia of Pharmaceutical Technology, eds. J.

[0983] Swarbrickand J. C. Boylan, 1988-1999, Marcel Dekker, New York. Proper formulation is dependent upon the route of administration chosen. The formulation and preparation of such compositions is well-known to those skilled in the art of pharmaceutical formulation. In preparing a formulation, the active compound can be milled to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

[0984] Dosages

[0985] The dosage of the compound used in the methods described herein, or pharmaceutically acceptable salts or prodrugs thereof, or pharmaceutical compositions thereof, can vary depending on many factors, e.g., the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The compounds used in the methods described herein may be administered initially in a ATTORNEY DOCKET NO.: D0963.70010WO00

[0986] suitable dosage that may be adjusted as required, depending on the clinical response. In some embodiments, a suitable daily dose of a compound of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. In some embodiments, an effective dose is a subtherapeutic dose relative to the monotherapy. Such an effective dose will generally depend upon the factors described above.

[0987] A compound of the invention may be administered to the patient in a single dose or in multiple doses. When multiple doses are administered, the doses may be separated from one another by, for example, 1-24 hours, 1-7 days, 1-4 weeks, or 1-12 months. The compound may be administered according to a schedule or the compound may be administered without a predetermined schedule. An active compound may be administered, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times per day, every 2nd, 3rd, 4th, 5th, or 6th day, 1, 2, 3, 4, 5, 6, or 7 times per week, 1, 2, 3, 4, 5, or 6 times per month, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times per year. It is to be understood that, for any particular subject, specific dosage regimes should be adjusted overtime according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions.

[0988] While the attending physician ultimately will decide the appropriate amount and dosage regimen, an effective amount of a compound of the invention may be, for example, a total daily dosage of, e.g., between 0.05 mg and 3000 mg of any of the compounds described herein.

[0989] Alternatively, the dosage amount can be calculated using the body weight of the patient. Such dose ranges may include, for example, between 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

[0990] In the methods of the invention, the time period during which multiple doses of a compound of the invention are administered to a patient can vary. For example, in some embodiments, doses of the compounds of the invention are administered to a patient over a time period that is 1-7 days; 1-12 weeks; or 1-3 months. In some embodiments, the compounds are administered to the patient over a time period that is, for example, 4-11 months or 1-30 years. In some embodiments, the compounds are administered to a patient at the onset of symptoms. In any of these embodiments, the amount of compound that is administered may vary during the time period of administration. When a compound is administered daily, administration may occur, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times per day.

[0991] Formulations

[0992] A compound identified as capable of treating any of the conditions described herein, using any of the methods described herein, may be administered to patients or animals with a pharmaceutically-acceptable diluent, carrier, or excipient, in unit dosage form. The chemical compounds for use in such therapies may be produced and isolated by any standard technique known to those in the field of medicinal chemistry. Conventional pharmaceutical practice may be employed to provide suitable formulations or compositions to administer the identified compound to ATTORNEY DOCKET NO.: D0963.70010WO00

[0993] patients suffering from a disease or condition. Administration may begin before the patient is symptomatic.

[0994] Exemplary routes of administration of the compounds (e.g., a compound of the invention), or pharmaceutical compositions thereof, used in the present invention include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intraorbital, intraventricular, intraspinal, intraperitoneal, intranasal, inhalation, and topical administration. The compounds desirably are administered with a pharmaceutically acceptable carrier. Pharmaceutical formulations of the compounds described herein formulated for treatment of the disorders described herein are also part of the present invention.

[0995] Formulations for Oral Administration

[0996] The pharmaceutical compositions contemplated by the invention include those formulated for oral administration (“oral dosage forms”). Oral dosage forms can be, for example, in the form of tablets, capsules, a liquid solution or suspension, a powder, or liquid or solid crystals, which contain the active ingredient(s) in a mixture with non-toxic pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates, or alginic acid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, carboxymethylcellulose sodium, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricating agents, glidants, and antiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc). Other pharmaceutically acceptable excipients can be colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[0997] Formulations for oral administration may also be presented as chewable tablets, as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules where the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil. Powders, granulates, and pellets may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g., a mixer, a fluid bed apparatus, or a spray drying equipment.

[0998] Controlled release compositions for oral use may be constructed to release the active drug by controlling the dissolution and / or the diffusion of the active drug substance. Any of a number of strategies can be pursued in order to obtain controlled release and the targeted plasma concentration versus time profile. In one example, controlled release is obtained by appropriate selection of various formulation parameters and ingredients, including, e.g., various types of controlled release compositions and coatings. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and ATTORNEY DOCKET NO.: D0963.70010WO00

[0999] liposomes. In some embodiments, compositions include biodegradable, pH, and / or temperaturesensitive polymer coatings.

[1000] Dissolution or diffusion-controlled release can be achieved by appropriate coating of a tablet, capsule, pellet, or granulate formulation of compounds, or by incorporating the compound into an appropriate matrix. A controlled release coating may include one or more of the coating substances mentioned above and / or, e.g., shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resins, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methylmethacrylate, 2-hydroxymethacrylate, methacrylate hydrogels, 1,3 butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycols. In a controlled release matrix formulation, the matrix material may also include, e.g., hydrated methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbon.

[1001] The liquid forms in which the compounds and compositions of the present invention can be incorporated for administration orally include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils, e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[1002] Formulations for Parenteral Administration

[1003] The compounds described herein for use in the methods of the invention can be administered in a pharmaceutically acceptable parenteral (e.g., intravenous or intramuscular) formulation as described herein. The pharmaceutical formulation may also be administered parenterally (intravenous, intramuscular, subcutaneous or the like) in dosage forms or formulations containing conventional, non-toxic pharmaceutically acceptable carriers and adjuvants. In particular, formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. For example, to prepare such a composition, the compounds of the invention may be dissolved or suspended in a parenterally acceptable liquid vehicle. Among acceptable vehicles and solvents that may be employed are water, water adjusted to a suitable pH by addition of an appropriate amount of hydrochloric acid, sodium hydroxide ora suitable buffer, 1,3-butanediol, Ringer’s solution and isotonic sodium chloride solution. The aqueous formulation may also contain one or more preservatives, for example, methyl, ethyl, or n-propyl p-hydroxybenzoate. Additional information regarding parenteral formulations can be found, for example, in the United States Pharmacopeia-National Formulary (USP-NF), herein incorporated by reference.

[1004] The parenteral formulation can be any of the five general types of preparations identified by the USP-NF as suitable for parenteral administration:

[1005] (1) Drug Injection: a liquid preparation that is a drug substance (e.g., a compound of the invention), or a solution thereof; ATTORNEY DOCKET NO.: D0963.70010WO00

[1006] (2) Drug for Injection: the drug substance (e.g., a compound of the invention) as a dry solid that will be combined with the appropriate sterile vehicle for parenteral administration as a drug injection;

[1007] (3) Drug Injectable Emulsion: a liquid preparation of the drug substance (e.g., a compound of the invention) that is dissolved or dispersed in a suitable emulsion medium;

[1008] (4) Drug Injectable Suspension: a liquid preparation of the drug substance (e.g., a compound of the invention) suspended in a suitable liquid medium; and

[1009] (5) Drug for Injectable Suspension: the drug substance (e.g., a compound of the invention) as a dry solid that will be combined with the appropriate sterile vehicle for parenteral administration as a drug injectable suspension.

[1010] Formulations for parenteral administration include solutions of the compound prepared in water suitably mixed with a surfactant, e.g., hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005) and in The United States Pharmacopeia: The National Formulary (USP 36 NF31), published in 2013.

[1011] Formulations for parenteral administration may, for example, contain excipients, sterile water, or saline, polyalkylene glycols, e.g., polyethylene glycol, oils of vegetable origin, or hydrogenated napthalenes. Biocompatible, biodegradable lactide polymer, lactide / glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be used to control the release of the compounds. Other potentially useful parenteral delivery systems for compounds include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Formulations for inhalation may contain excipients, for example, lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or may be oily solutions for administration in the form of nasal drops, or as a gel.

[1012] The parenteral formulation can be formulated for prompt release or for sustained / extended release of the compound. Exemplary formulations for parenteral release of the compound include: aqueous solutions, powders for reconstitution, cosolvent solutions, oil / water emulsions, suspensions, oil-based solutions, liposomes, microspheres, and polymeric gels.

[1013] Example 1: Investigating Cancer Patients with Hotspot Missense and Biallelic Truncating Mutations

[1014] The present example is based on data (e.g., Cyclin E expression data and clinical outcome data) obtained from patients enrolled in National Clinical Trial (NCT) number: NCT04855656 (“MYTHIC”) who were treated with lunresertib (RP-6306) in combination with camonsertib (RP-3500).

[1015] Cyclin E immunohistochemistry (IHC) was performed on patient tissue samples collected from the MYTHIC trial and compared with IHC on tumor tissue biobank samples unselected for lunresertib biomarkers; this revealed that high levels of Cyclin E (e.g., Cyclin E H-Score >100) accumulate when ATTORNEY DOCKET NO.: D0963.70010WO00

[1016] F-box / WD repeat containing protein (FBXW7) mutations are present (FIGs. 1A-B). Notably, three common arginine (R) hotspot mutations (e.g., R465, R479, and R505) in the FBXW7 protein occurred at a high frequency in patients enrolled in MYTHIC (FIG. 2). These hotspot mutations occurred within the third and fourth WD40 repeats that disrupt recognition of the Cyclin E1 substrate and are deleterious, missense mutations. Additional mutations, such as truncating mutations, inframe mutations, and splice site mutations that span FBXW7 were also identified (FIG. 2).

[1017] All identified mutations were next stratified into three groups (e.g., high confidence, low confidence, or unknown) based on FBXW7 loss-of-function (LoF) quality. High confidence FBXW7 mutations included hotspots mutations (e.g., R465, R479, R505) and biallelic truncations. Low confidence FBXW7 mutations included monoallelic truncations, missense, and splice mutations. Nonhotspot mutations where allelic status is indeterminate, or biallelic missense and splice mutations were considered “unknown.” Notably, the hotspot and biallelic truncation mutations coincided with the greatest accumulation of Cyclin E (FIG. 3). Overall, colorectal cancer (CRC) patients in MYTHIC having high confidence LoF mutations (i.e., hotspot and biallelic truncation mutations) tend to stay on the combination therapy (e.g., lunresertib + camonsertib) longer than patients with low confidence FBXW7 LoF mutations. The numerical values of Cyclin E show that the high confidence FBXW7 mutations are also associated with higher Cyclin E (e.g., Cyclin E H-Score > 100) (FIG.4).

[1018] Table 4 below summarizes observed FBXW7 mutations, Cyclin E expression, and clinical outcomes in CRC patients treated with lunresertib (RP-6306) in combination with camonsertib (RP- 3500) in the MYTHIC trial. Mutations and variant allele frequencies (VAFs) are as reported in local Next Generation Sequencing (NGS) test reports. Allelic status (AS) as assessed using the SyNthetic lethal Interactions for Precision Diagnostics (SNiPDx) assay is reported where available. Cyclin E H- score was determined by IHC. FBXW7 quality was manually curated based on totality of available evidence from local NGS and SNiPDx.

[1019] Table 4. Exemplary FBXW7 Mutations Identified in MYTHIC

[1020] FBXW7

[1021] Cyclin RP- RP- Mutation FBXW7 Patient DoT Patient FBXW7 E 6306 3500 BOR PFS (VAF, quality

[1022] ID (Wks) Status quality H- dose dose Allelic reason score status)

[1023] 2001- 240 80 PR 27.10 27.57 Off-tx P. R505C High Hotspot 160 0108 QD QD (11.2%,

[1024] 3 / 4 3 / 4; mono)

[1025] co nt

[1026] 1002- 80 80 SD 24.60 25.14 Off-tx P. R465H High Hotspot 150 0134 BID QD (unk,

[1027] 3 / 4 3 / 4; unk);

[1028] 2 / 1 p. R224*

[1029] (unk,

[1030] unk)

[1031] 1001- 60 80 SD 11.70 20.14 Off-tx P. R479Q High Hotspot 180 0081 BID QD (unk,

[1032] 3 / 4 3 / 4; unk)

[1033]

[1034] co nt ATTORNEY DOCKET NO.: D0963.70010WO00

[1035] FBXW7 Cyclin Patient RP- RP- DoT Patient Mutation FBXW7 FBXW7

[1036] 6306 3500 BOR PFS (VAF, quality E ID (Wks) Status quality H- dose dose Allelic reason score status)

[1037] 1008- 80 80 SD 19.30 19.29 Off-tx P. R465C High Hotspot 90 0152 BID (23.6%, QD

[1038] 3 / 4 3 / 4; mono)

[1039] co nt

[1040] 4001- 80 80 SD 11.30 16.29 Off-tx P. R465H High Hotspot 1 0207 BID QD (51.8%,

[1041] 3 / 4 3 / 4; unk)

[1042] co nt

[1043] 1008- 80 80 PD 5.70 14.00 Off-tx P. R465H High Hotspot NA 0205 BID QD (8.5%,

[1044] 3 / 4 3 / 4; unk)

[1045] co nt

[1046] 1008- 120 80 PD 12.00 12.14 Off-tx P. Q477S High Hotspot 75 0056 QD QD (VUS)

[1047] co nt 3 / 4; (11.7%,

[1048] co nt unk);

[1049] P. R465C

[1050] (10%,

[1051] unk)

[1052] 1011- 120 120 PD 5.10 12.14 Off-tx P. W425C Low Missense, 25 0076 QD QD (40%, monoallelic

[1053] co nt 3 / 4; mono)

[1054] 2 / 1

[1055] 4001- 40 80 PD 5.30 12.14 Off-tx P. R689W Low Missense, 9 0101 BID QD (31.9%, monoallelic

[1056] co nt 3 / 4; mono)

[1057] co nt

[1058] 1004- 60 80 PD 5.90 9.57 Off-tx p. R393* Low Truncation, 125 0168 BID QD (27.2%, monoallelic 3 / 4 3 / 4; mono)

[1059] co nt

[1060] 4001- 80 80 PD 6.10 6.43 Off-tx p. S546* Unk Truncation, NA 0245 BID QD (30.1%, unknown

[1061] 3 / 4 3 / 4; unk) AS

[1062] co nt

[1063] 1001- 120 80 PD 6.10 6.29 Off-tx P. G477S Unk Missense 90 0055 QD QD (unk, unknown

[1064] co nt 3 / 4; unk) AS

[1065] co nt

[1066] 1004- 80 80 PD 5.10 6.14 Off-tx p. R479* Unk Truncation, 1 0117 BID QD (unk, unknown

[1067] 3 / 4 3 / 4; unk) AS

[1068] co nt

[1069] 1001- 80 80 NE 0.14 5.86 Off-tx P. R465H High Hotspot 0 0218 BID QD (0.5%,

[1070] 3 / 4 3 / 4; unk)

[1071] co nt

[1072] 1004- 80 80 PD 5.10 5.57 Off-tx P. R465H High Hotspot 0 0208 BID QD (14.8%,

[1073] 3 / 4 3 / 4; unk)

[1074] co nt

[1075]

[1076] RP-6306 = lunresertib; RP-3500 = camonsertib; BOR = best overall response; DoT = duration of treatment; Wks = weeks; FBXW7 = F-box / WD repeat containing protein; VAF = variant allele frequency. ATTORNEY DOCKET NO.: D0963.70010WO00

[1077] FBXW7 mutations were also studied in vitro using retinal RPE1 cells and colorectal DLD1 cells. As shown in FIG 5, Cyclin E1 accumulated in retinal RPE1 (FIGs. 5A-B) and colorectal DLD1 (FIGs.5D-E) cell lines following FBXW7 truncation by clustered regularly interspaced short palindromic repeats (CRISPR)-mediated base editing (CRISPR-BE), relative to control guide (sgPLK1silent). Notably, such FBXW7 truncations also showed an increased sensitivity to lunresertib (RP-6306) treatment in the RPE1 (FIG.5C) and DLD1 (FIG.5D) cell lines. Biallelic status of these truncations were independently confirmed (data not shown).

[1078] Taken together, these data show that hotspot FBXW7 missense and biallelic truncating mutations cause true LoF, as illustrated in FIG. 6.

Claims

ATTORNEY DOCKET NO.: D0963.70010WO00CLAIMS1. A method for treating a cancer in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of a membrane-associated tyrosine and threonine-specific cdc2-inhibatory kinase (Myt1) inhibitor, wherein the cancer comprises a truncating biallelic loss-of-function mutation in FBXW7.

2. A method for treating a cancer in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having a truncating biallelic loss-of-function mutation in FBXW7.

3. A method of inducing cell death in a cancer cell having a truncating biallelic loss-of-function mutation in FBXW7, the method comprising contacting the cell with an effective amount of a Myt1 inhibitor.

4. A method of inducing cell death in a cancer cell, the method comprising administering to a subject in need thereof an effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as having a truncating biallelic loss-of-function mutation in FBXW7.

5. The method of claim 1 or 3, wherein the method further comprises identifying the cancer or cancer cell as having the truncating biallelic loss-of-function mutation in FBXW7.

6. The method of any one of claims 1 to 5, wherein the truncating biallelic loss-of-function mutation comprises one or two truncating mutations selected from the group consisting of a nonsense mutation, a frameshift mutation, and a splice site mutation.

7. The method of claim 6, wherein the truncating mutation produces an FBXW7 transcript that is 50-1800 nucleotides in length.

8. The method of any one of claims 1 -7, wherein the truncating biallelic loss-of-function mutation comprises one truncating mutation in a first allele of FBXW7 and (i) at least one mutation in a second allele of FBXW7; (ii) a partial deletion of a second allele of FBXW7; or (iii) a complete deletion of a second allele of FBXW7.

9. The method of claim 8, wherein the at least one mutation in the second allele of FBXW7 is a missense mutation.ATTORNEY DOCKET NO.: D0963.70010WO0010. The method of any one of claims 1 to 9, wherein the cancer is colorectal cancer (CRC), uterine cancer, bladder cancer, cervical cancer, and upper gastrointestinal cancer.

11. The method of any one of claims 1 to 10, wherein the cancer is CRC.

12. The method of any one of claims 1 to 11, wherein the cancer is further characterized by CCNE1 overexpression.

13. The method of claim 12, wherein the method further comprises identifying the cancer as having CCNE1 overexpression.

14. The method of claim 5, wherein identifying the cancer or cancer cell as having the truncating biallelic loss-of-function mutation in FBXW7 comprises targeted panel sequencing, whole genome sequencing (WGS) or Whole Exome Sequencing (WES).

15. The method of any one of claims 1 to 14 further comprising administration of a therapeutically effective amount of a Wee1 inhibitor, Fen1 inhibitor, Topi inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Aurkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod1 inhibitor, Sod2 inhibitor, Bub1 inhibitor, Cdc7 inhibitor, Sae1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Dut inhibitor, Hdac3 inhibitor, Chekl inhibitor, Aurka inhibitor, Men1 inhibitor, Doti I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinum-based DNA damaging agent, or a combination thereof.

16. The method of any one of claims 1 to 15, wherein the method comprises administering the Wee1 inhibitor.

17. The method of claim 16, wherein the Wee1 inhibitor is one of:ATTORNEY DOCKET NO.: D0963.70010WO0018. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Fen1 inhibitor.

19. The method of claim 18, wherein the Fen1 inhibitor is C8 (PMID: 32719125), SC13, Fen1-IN-3, or a pharmaceutically acceptable salt thereof.

20. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Topi inhibitor.

21. The method of claim 20, wherein the Topi inhibitor is irinotecan, topotecan, camptothecin, lamellarin D, or a pharmaceutically acceptable salt thereof.

22. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Rrm1 inhibitor.

23. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Rrm2 inhibitor.

24. The method of claim 23, wherein the Rrm2 inhibitor is motexafin gadolinium, hydroxyurea, fludarabine, cladribine, tezacitabine, triapine, or a pharmaceutically acceptable salt thereof.

25. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Aurkb inhibitor.

26. The method of claim 25, wherein the Aurkb inhibitor is MK0547, AZD1152, PHA739358, AT9283, AMG900, SNS-314, TAK-901, 2CYC116, GSK1070916, PF03814735, or a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0027. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Top2A inhibitor.

28. The method of claim 43, wherein the Top2A inhibitor is etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, or a pharmaceutically acceptable salt thereof.

29. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the ATR inhibitor.

30. The method of claim 29, wherein the ATR inhibitor is a compound of formula (IV):Y-Y(IV),or a pharmaceutically acceptable salt thereof,wherein- is a double bond, and each Y is independently N or CR4; or - is a single bond, and each Y is independently NRY, carbonyl, or C(RY)2; wherein each RYis independently H or optionally substituted Ci-e alkyl;R1is optionally substituted Ci-e alkyl or H;R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2,-SO2R5A, or-Q-R5B;R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl; each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl;each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-aryl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, or-SO2R5A; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;ATTORNEY DOCKET NO.: D0963.70010WO00each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted Ce- aryl;R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or optionally substituted alkoxy;each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted Ce- arylene; andX is hydrogen or halogen.

31. The method of claim 29, wherein the ATR inhibitor is a compound of formula (V):N^R1(V),or a pharmaceutically acceptable salt thereof,whereineach Y is independently N or CR4;R1is optionally substituted C1-6 alkyl or H;R2is optionally substituted C2-9 heterocyclyl, optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce-aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R5)2, -OR5, -CON(R6)2, -SO2N(R6)2,-SO2R5A, or-Q-R5B;R3is optionally substituted C1-9 heteroaryl or optionally substituted C1-9 heteroaryl C1-6 alkyl; each R4is independently hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, or optionally substituted C2-6 alkynyl;each R5is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-aryl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, or-SO2R5A; or both R5, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;each R5Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted Ce- aryl;ATTORNEY DOCKET NO.: D0963.70010WO00R5Bis hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, -N(R5)2, -CON(R6)2, -SO2N(R6)2, -SO2R5A, or optionally substituted alkoxy;each R6is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or both R6, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl;Q is optionally substituted C2-9 heterocyclylene, optionally substituted C3-8 cycloalkylene, optionally substituted C1-9 heteroarylene, or optionally substituted Ce- arylene; andX is hydrogen or halogen.

32. The method of any one of claim 29 to 31, wherein the ATR inhibitor is selected from the group consisting of compounds A43, A57, A62, A87, A93, A94, A95, A99, A100, A106, A107, A108, A109, A111, A112, A113, A114, A115, A116, A118, A119, A120, A121 (camonsertib), A122, A123, A135, A147, A148, and pharmaceutically acceptable salts thereof.

33. The method of claim 32, wherein the ATR inhibitor is compound A43 or a pharmaceutically acceptable salt thereof.

34. The method of claim 32, wherein the ATR inhibitor is camonsertib or a pharmaceutically acceptable salt thereof.

35. The method of claim 32, wherein the ATR inhibitor is compound A122 or a pharmaceutically acceptable salt thereof.

36. The method of claim 29 wherein the ATR inhibitor isHl / elimusertib (BAY1895344) ceralasertib (AZD6738) berzosertib (VE-822)ATTORNEY DOCKET NO.: D0963.70010WO00Gartisertib (VX-803, M4344) tuvusertib (M1774)SKLB-197or a pharmaceutically acceptable salt thereof.

37. The method claim 15, wherein the method comprises the step of administering the Ttk inhibitor.

38. The method of claim 37, wherein the Ttk inhibitor is BAY1217389 or a pharmaceutically acceptable salt thereof.

39. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Sod1 inhibitor.

40. The method of claim 39, wherein the Sod1 inhibitor is LCS1, ATN-224, Pyrimethamine, a compound of the following structureor a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0041. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Sod2 inhibitor.

42. The method of claim 41, wherein the Sod2 inhibitor is LCS1, ATN-224, pyrimethamine, or a pharmaceutically acceptable salt thereof.

43. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Bub1 inhibitor.

44. The method of claim 43, wherein the Bub1 inhibitor is BAY-320, BAY-419, BAY1816032, or a pharmaceutically acceptable salt thereof.

45. The method of claim 15, wherein the method comprises the step of administering the Cdc7 inhibitor.

46. The method of claim 45, wherein the Cdc7 inhibitor is SRA141, TAK931, or a pharmaceutically acceptable salt thereof.

47. The method of claim 46, wherein the method comprises the step of administering the Sae1 inhibitor.

48. The method of claim 47, wherein the Sae1 inhibitor is ML792 or a pharmaceutically acceptable salt thereof.

49. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Plk1 inhibitor.

50. The method of claim 49, wherein the Plk1 inhibitor is BI2536, BI6727, TAK960, NMSP937, GSK461364, or a pharmaceutically acceptable salt thereof.

51. The method of claim 15, wherein the method comprises the step of administering the Uba2 inhibitor.

52. The method of claim 51, wherein the Uba2 inhibitor is TAK981 or a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0053. The method of any one of claims 15, wherein the method comprises the step of administering the Dut inhibitor.

54. The method of claim 53, wherein the Dut inhibitor is TAS114 or a pharmaceutically acceptable salt thereof.

55. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Hdac3 inhibitor.

56. The method of claim 55, wherein the Hdac3 inhibitor is RGFP966 or a pharmaceutically acceptable salt thereof.

57. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Chekl inhibitor.

58. The method of claim 57, wherein the Chekl inhibitor is SRA737 or a pharmaceutically acceptable salt thereof.

59. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Aurka inhibitor.

60. The method of claim 59, wherein the Aurka inhibitor is MLN8237, MK0547, MLN8054, PHA739358, AT9283, AMG900, MK5108, SNS314, TAK901, CYC116, ENMD2076, or a pharmaceutically acceptable salt thereof.

61. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Men1 inhibitor.

62. The method of claim 61, wherein the Men1 inhibitor is MI3454, SNDX5613, VTP50469, KO539, or a pharmaceutically acceptable salt thereof.

63. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Doti I inhibitor.

64. The method of claim 63, wherein the Doti I inhibitor is EPZ5676 or a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0065. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Crebbp inhibitor.

66. The method of claim 65, wherein the Crebbp inhibitor is CPI4, CCS1477, E7386, NEO1132, NEO2734, PRI724, C82, BC001, C646, EML425, CBP30, or a pharmaceutically acceptable salt thereof.

67. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Ezh2 inhibitor.

68. The method of claim 67, wherein the Ezh2 inhibitor is EPZ-6438, GSK126, or a pharmaceutically acceptable salt thereof.

69. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Plk4 inhibitor.

70. The method of claim 69, wherein the Plk4 inhibitor is a compound of formula (VI)or a pharmaceutically acceptable salt thereof,whereinn is 0, 1, 2, 3, or 4;m is 0, 1, or 2;L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted Ce- aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;R1bis hydrogen; orR1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;ATTORNEY DOCKET NO.: D0963.70010WO00A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor-N(R3B)2each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;X is N, and R4is absent; orX is C, and R4is hydrogen, halogen, cyano, optionally substituted amino, optionally substituted acyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted Ce- arylene, or optionally substituted C3-8 cycloalkylene;R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substitutedATTORNEY DOCKET NO.: D0963.70010WO00C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; andR6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

71. The method of claim 69 or claim 70, wherein the Plk4 inhibitor is a compound of formula (VII)or a pharmaceutically acceptable salt thereof,whereinn is 0, 1, 2, 3, or 4;m is 0, 1, or 2;L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted Ce- aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;R1bis hydrogen; orR1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl,ATTORNEY DOCKET NO.: D0963.70010WO00optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor-N(R3B)2each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted Ce- arylene, or optionally substituted C3-8 cycloalkylene;R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; andR6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

72. The method of claim 69 or claim 70, wherein the Plk4 inhibitor is a compound of formula (VIII)ATTORNEY DOCKET NO.: D0963.70010WO00R6(VIII)or a pharmaceutically acceptable salt thereof,whereinn is 0, 1, 2, 3, or 4;m is 0, 1, or 2;L is optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heteroaryl, optionally substituted Ce- aryl, or optionally substituted C3-8 cycloalkyl, wherein L is further optionally substituted by n occurrences of R3;R1ais hydrogen, halogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkoxy, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or nitrile;R1bis hydrogen; orR1aand R1b, together with the atoms to which they are attached, are a 3-5-membered cycloalkyl, cycloakylene, cycloalkylyne, heterocycloalkyl, aryl, or heteroaryl;A is O or S, and R2Aand R2Bare both absent; or A is N, R2Ais absent, and R2Bis hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl, or R2Band L, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl or optionally substituted C2-9 heteroaryl; or A is C, and each of R2Aand R2Bare independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; each R3is independently halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, -S(O)mR3A, -N(R3B)2, or -OR3B;R3Ais optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl,ATTORNEY DOCKET NO.: D0963.70010WO00optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -OR3Bor-N(R3B)2each R3Bis independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or optionally substituted C1-6 alkylsulfonyl; or two R3Bgroups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;R4is hydrogen, halogen, cyano, optionally substituted amino, optionally substituted acyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;R5is optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -CONH2, or-Z-R5A;Z is optionally substituted amino, optionally substituted C2-9 heterocyclylene, optionally substituted C2-9 heteroarylene, optionally substituted Ce- arylene, or optionally substituted C3-8 cycloalkylene;R5Ais hydrogen, halogen, cyano, optionally substituted C1-6 alkylsulfonyl, optionally substituted C1-6 alkyl, optionally substituted C1-6 heteroalkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, or optionally substituted C1-9 heteroaryl;R6is hydrogen, halogen, cyano, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, or -OR6A; andR6Ais hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, or optionally substituted C3-8 cycloalkyl.

73. The method of claim 69 or claim 70, wherein the Plk4 inhibitor is selected from the group consisting of compound P1 to P328.

74. The method of claim 73, wherein the Plk4 inhibitor is selected from the group consisting of compounds P16, P17, P25, P44, P45, P62, P110, P121, P137, P140, P151, P156, P164, P174, P179, P182, P183, P188, P195, P196, P197, P210, P189, P232, P238, P240, P243, P249, P252, 255, P260, P262, P272, P273, P274, P276, P277, P280, P308, P328, P334, P340, P343, P347, P348, P349, P350, P351, P352, P353, P354, P355, P356, P357, P358, P359, P360, P361, P362, P363, and P364.ATTORNEY DOCKET NO.: D0963.70010WO0075. The method of claim 69, wherein the Plk4 inhibitor is centrinone, CFI400945, or a pharmaceutically acceptable salt thereof.

76. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Haspin inhibitor.

77. The method of claim 76, wherein the Haspin inhibitor is SEL120 or a pharmaceutically acceptable salt thereof.

78. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the Mettl3 inhibitor.

79. The method of claim 78, wherein the Mettl3 inhibitor is UZH1 a, sTC-15, or a pharmaceutically acceptable salt thereof.

80. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the nucleoside analog.

81. The method of claim 80, wherein the nucleoside analog is cytarabine, gemcitabine, mercaptopurine, azacytidine, cladribine, decitabine, fluorouracil, floxuridine, fludarabine, nelarabine, or a pharmaceutically acceptable salt thereof or a combination thereof.

82. The method of claim 80 or claim 81, wherein the nucleoside analog is gemcitabine or a pharmaceutically acceptable salt thereof.

83. The method of claim 80 or claim 81, wherein the nucleoside analog is fluorouracil or a pharmaceutically acceptable salt thereof.

84. The method of claim 80 or claim 81, wherein the nucleoside analog is a combination of gemcitabine or a pharmaceutically acceptable salt thereof and fluorouracil or a pharmaceutically acceptable salt thereof.

85. The method of any one of claims 1 to 15, wherein the method comprises the step of administering the platinum-based DNA damaging agent.

86. The method of claim 85, wherein the platinum-based DNA damaging agent is cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, orsatraplatin.ATTORNEY DOCKET NO.: D0963.70010WO0087. The method of claim 85, wherein the platinum-based DNA damaging agent is carboplatin.

88. The method of any one of claims 15 to 87, wherein the Wee1 inhibitor, Fen1 inhibitor, Topi inhibitor, Rrm1 inhibitor, Rrm2 inhibitor, Au rkb inhibitor, Top2A inhibitor, ATR inhibitor, Ttk inhibitor, Sod1 inhibitor, Sod2 inhibitor, Bub1 inhibitor, Cdc7 inhibitor, Sae1 inhibitor, Plk1 inhibitor, Uba2 inhibitor, Dut inhibitor, Hdac3 inhibitor, Chekl inhibitor, Au rka inhibitor, Men1 inhibitor, Doti I inhibitor, Crebbp inhibitor, Ezh2 inhibitor, Plk4 inhibitor, Haspin inhibitor, Mettl3 inhibitor, nucleoside analog, platinum-based DNA damaging agent, or the combination thereof is administered as a pharmaceutical composition.

89. The method of any one of claims 1 to 88, wherein the Myt1 inhibitor is a compound of formula (I):or a pharmaceutically acceptable salt thereof,whereineach of X, Y, and Z is independently N or CR2;R1and each R2are independently hydrogen, optionally substituted Ci-e alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, cyano, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or -Q-R7B; or R1combines with one R2that is vicinal to R1to form an optionally substituted C3-6 alkylene;each of R3and R4is independently optionally substituted C1-6 alkyl or halogen;R5is H or -N(R7)2;R6is -C(O)NH(R8), -C(O)R7A, or -SO2R7A;each R7is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted Ce-aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted Ce- aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or -SO2R7A; or two R7groups, together with the atom to which both are attached, combine to form an optionally substituted C2-9 heterocyclyl;ATTORNEY DOCKET NO.: D0963.70010WO00each R7Ais independently optionally substituted C1-6 alkyl, optionally substituted C3-8 cycloalkyl, or optionally substituted Ce- aryl;each R7Bis independently hydroxyl, optionally substituted C1-6 alkyl, optionally substituted Ce-aryl, optionally substituted C2-9 heterocyclyl, optionally substituted C1-9 heteroaryl, -N(R7)2, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or optionally substituted alkoxy;each R8is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted Ce- aryl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or two R8, together with the atom to which they are attached, combine to form an optionally substituted C2-9 heterocyclyl; andQ is optionally substituted C1-6 alkylene, optionally substituted C2-6 alkenylene, optionally substituted C2-6 alkynylene, optionally substituted C3-8 cycloalkylene, optionally substituted C3-8 cycloalkenylene optionally substituted Ce- arylene, optionally substituted C2-9 heterocyclylene, or optionally substituted C1-9 heteroarylene.

90. The method of claim 89, wherein the compound is enriched for the atropisomer of formula (IA):(IA),or a pharmaceutically acceptable salt thereof.

91. The method of claim 89 or 90, wherein X is CR2.

92. The method of claim 90, wherein the compound is of formula (II):or a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0093. The method of claim 92, wherein the compound is enriched for the atropisomer of formula (HA):(HA);or a pharmaceutically acceptable salt thereof.

94. The method of claim 89, wherein the compound is of formula (III):wherein R2Ais hydrogen, optionally substituted Ci-e alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclyl, optionally substituted C2-9 heterocyclyl C1-6 alkyl, optionally substituted Ce- aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, -N(R7)2, -OR7, -C(O)N(R8)2, -SO2N(R8)2, -SO2R7A, or-Q-R7B;or a pharmaceutically acceptable salt thereof.

95. The method of claim 94, wherein the compound is enriched for the atropisomer of formula (III A):(IIIA);or a pharmaceutically acceptable salt thereof.ATTORNEY DOCKET NO.: D0963.70010WO0096. The method of claim 94 or 95, wherein R2Ais hydrogen, optionally substituted Ci-e alkyl, or halogen.

97. The method of any one of claims 89 to 96, wherein R3is optionally substituted Ci-e alkyl.

98. The method of any one of claims 89 to 96, wherein R3is halogen.

99. The method of claim 98, wherein the halogen is chlorine.

100. The method of any one of claims 89 to 99, wherein R4is optionally substituted Ci-e alkyl.

101. The method of any one of claims 89 to 99, wherein R4is halogen.

102. The method of claim 101, wherein the halogen is chlorine.

103. The method of any one of claims 89 to 102, wherein R2is hydrogen.

104. The method of any one of claims 89 to 102, wherein R2is optionally substituted Ci-e alkyl.

105. The method of claim 104, wherein R2is optionally substituted methyl or optionally substituted isopropyl.

106. The method of any one of claims 89 to 102, wherein R2is halogen.

107. The method of any one of claims 89 to 106, wherein R1is hydrogen.

108. The method of any one of claims 89 to 106, wherein R1is halogen.

109. The method of claim 108, wherein R1is chlorine or bromine.

110. The method of any one of claims 89 to 106, wherein R1is optionally substituted Ci-e alkyl.

111. The method of claim 110, wherein R1is optionally substituted methyl, optionally substituted ethyl, optionally substituted isopropyl, or optionally substituted butyl.

112. The method of any one of claims 89 to 106, wherein R1is optionally substituted C1-9 heteroaryl.ATTORNEY DOCKET NO.: D0963.70010WO00113. The method of claim 112, wherein R1is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, or pyrazolyl, wherein R1is optionally substituted with substituents as defined for optionally substituted C1-9 heteroaryl.

114. The method of any one of claims 89 to 106, wherein R1is optionally substituted C3-8 cycloalkyl.

115. The method of claim 114, wherein R1is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, wherein R1is optionally substituted with substituents as defined for optionally substituted C3-8 cycloalkyl.

116. The method of any one of claims 89 to 106, wherein R1is optionally substituted C2-9 heterocyclyl.

117. The method of claim 116, wherein R1is 1,2,3,6-tetrahydropyridinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, oxa-aza-spiro[3,3]heptane, or oxa-aza-bicyclo[3.2.1]octane, wherein R1is optionally substituted with substituents as defined for optionally substituted C2-9 heterocyclyl.

118. The method of any one of claims 89 to 106, wherein R1is optionally substituted C3-8 cycloalkyl.

119. The method of claim 118, wherein R1is optionally substituted cyclohexenyl or optionally substituted cyclopentenyl.

120. The method of any one of claims 89 to 106, wherein R1is optionally substituted Ce- aryl.

121. The method of claim 120, wherein R1is optionally substituted phenyl.

122. The method of any one of claims 89 to 106, wherein R1is -Q-R7B.

123. The method of claim 122, wherein Q is optionally substituted C2-6 alkynylene.

124. The method of claim 122, wherein Q is optionally substituted C1-6 alkylene.ATTORNEY DOCKET NO.: D0963.70010WO00125. The method of claim 122, wherein Q is optionally substituted Ce- arylene.

126. The method of any one of claims 122 to 125, wherein R7Bis optionally substituted C2-9 heterocyclyl.

127. The method of any one of claims 122 to 125, wherein R7Bis optionally substituted Ce-aryl.

128. The method of any one of claims 89 to 127, wherein R1is optionally substituted with one, two, or three groups independently selected from the group consisting of methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, oxo, -C(O)NH2, -C(O)NH(Me), -C(O)N(Me)2, -(CH2)n-C(O)OH, and -(CH2)n-C(O)Ot-Bu, wherein n is 0 or 1.

129. The method of any one of claims 89 to 106, wherein R1is -N(R7)2.

130. The method of claim 129, wherein R1is diethylamino.

131. The method of any one of claims 89 to 130, wherein R5is hydrogen.

132. The method of any one of claims 89 to 130, wherein R5is -N(R7)2.

133. The method of claim 132, wherein R5is -NH2.

134. The method of any one of claims 89 to 133, wherein R6is -C(O)NH(R8).

135. The method of any one of claims 89 to 134, wherein R6is -C(O)NH2.

136. The method of any one of claims 89 to 134, wherein R6is -C(O)NH(Me).

137. The method of any one of claims 89 to 133, wherein R6is -SO2R7A.

138. The method of claim 137, wherein R6is -SO2Me.

139. The method of any one of claims 1 to 88, wherein the Myt1 inhibitor is selected from the group consisting of compounds 1-328 and pharmaceutically acceptable salts thereof.ATTORNEY DOCKET NO.: D0963.70010WO00140. The method of any one of claims 1 to 139, wherein the Myt1 inhibitor is administered as a pharmaceutical composition.

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