Cyclin-dependent kinase 2 biomarkers and their use

Abstract

Biomarkers are provided that predict and / or indicate a subject's responsiveness to a cyclin-dependent kinase 2 (CDK2) inhibitor. The biomarkers, compositions, and methods described herein are useful for selecting appropriate treatment modalities and monitoring treatment for subjects who have, are suspected of having, or are at risk of developing a disease or disorder associated with CDK2.

Description

[Technical Field]

[0001] Related applications This application claims the interests of U.S. Provisional Patent Application No. 62 / 806,265, filed on 15 February 2019, which is incorporated herein by reference in its entirety.

[0002] Sequence List This application includes an electronically submitted sequence listing in ASCII format, the entirety of which is incorporated herein by reference. The ASCII copy, created on January 23, 2020, is named 20443-0588WO1_SL.txt and is 15,865 bytes in size.

[0003] This invention generally relates to biomarkers and cancer. [Background technology]

[0004] Cyclin-dependent kinases ("CDKs") are a family of serine / threonine kinases. When CDKs heterodimerized with regulatory subunits known as cyclins are fully activated, they become the driving force behind the cell cycle and cell division. Uncontrolled proliferation is a prominent feature of cancer cells, and misregistration of CDK function occurs frequently in many tumors. CDK2 and CDK4 are of particular interest because their activity is frequently dysregulated in a wide variety of human cancers. Therefore, CDKs are recognized as attractive targets for the design and development of compounds that can specifically bind to and inhibit the activity of CDKs in cancer cells, thus potentially functioning as therapeutic agents. Potent and highly selective CDK4 / 6 inhibitors, palbociclib, abemaciclib, and ribociclib, have been developed and approved by the U.S. Food and Drug Administration ("FDA") for the treatment of ER+ advanced breast tumors. Despite considerable effort, there are currently no FDA-approved drugs targeting CDK2. The lack of biomarkers that reliably report the enzymatic and / or oncogenic activity of CDK2 has hindered the development of effective target-associated assays for advanced discovery and optimization. There is a clear need to identify biomarkers for CDK2-mediated carcinogenesis to provide rapid and effective means for the development and evaluation of CDK2-targeted anticancer therapies. [Overview of the project]

[0005] The present invention is at least partially based on the discovery that the functional state of cyclin-dependent kinase inhibitor 2A (also known as "CDKN2A" or "p16") is a biomarker for predicting sensitivity to CDK2-targeted therapy in G1 / S-specific cyclin E1- ("CCNE1-")-amplified cells, which is suitable for use in patient stratification. In addition, the present invention is at least partially based on the discovery that in CCNE1-amplified cell lines, the phosphorylation level of human retinoblastoma-associated protein ("Rb") at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is a pharmacodynamic marker of CDK2 activity and is suitable for use in cell assays or preclinical and clinical applications, such as monitoring the course of treatment with CDK2 inhibitors or the response to treatment.

[0006] This disclosure features a method for treating human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, the method comprising administering a CDK2 inhibitor to the human subject, the human subject having been determined to have (i)(a) a nucleotide sequence encoding the p16 protein including the amino acid sequence of SEQ ID NO: 1, (b) a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or (c) expressing the p16 protein, and (ii)(a) amplification of the CCNE1 gene, and / or (b) a CCNE1 expression level higher than the control expression level of CCNE1 in a biological sample obtained from the human subject. In some embodiments, the subject has a CDK2-related disease or disorder. In some embodiments, the subject is suspected of having, or is at risk of developing, a CDK2-related disease or disorder. In some embodiments, the human subject has been determined to have (i)(a) a nucleotide sequence encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1, and / or (b) a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and (ii) amplification of the CCNE1 gene in a biological sample obtained from the human subject. In some embodiments, the CDKN2A gene encodes a protein containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the second therapeutic agent is administered to the human subject in combination with a CDK2 inhibitor. In some embodiments, the second therapeutic agent is a BCL2 inhibitor or a CDK4 / 6 inhibitor.

[0007] This disclosure also features a method for treating human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, the method comprising (i) identifying the presence of (a) a nucleotide sequence encoding the p16 protein, including the amino acid sequence of SEQ ID NO: 1, (b) a CDKN2A gene lacking one or more inactivating nucleic acid substitutions, and / or (c) the p16 protein in a biological sample obtained from a human subject; (ii) identifying (a) amplification of the CCNE1 gene and / or (b) an expression level of CCNE1 higher than the control expression level of CCNE1 in a biological sample obtained from a human subject; and (iii) administering a CDK2 inhibitor to the human subject. In some embodiments, the subject has a CDK2-related disease or disorder. In some embodiments, the subject is suspected of having, or is at risk of developing, a CDK2-related disease or disorder. In some embodiments, the method includes (i) identifying the presence of (a) a nucleotide sequence encoding the p16 protein, including the amino acid sequence of SEQ ID NO: 1, (b) a CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) the p16 protein in a biological sample obtained from a human subject, (ii) identifying amplification of (a) the CCNE1 gene in a biological sample obtained from a human subject, and (iii) administering a CDK2 inhibitor to the human subject. In some embodiments, the CDKN2A gene encodes a protein including the amino acid sequence of SEQ ID NO: 1. In some embodiments, a second therapeutic agent is administered to the human subject in combination with a CDK2 inhibitor. In some embodiments, the second therapeutic agent is a BCL2 inhibitor or a CDK4 / 6 inhibitor.

[0008] This disclosure also features a method for predicting the response to CDK2 inhibitors in human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, the method comprising (i) determining from a biological sample obtained from a human subject (a) the nucleotide sequence of the CDKN2A gene, (b) the presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) the presence of the p16 protein, and (ii) determining from a biological sample obtained from a human subject (a) the CCNE1 gene The process involves determining the copy number of offspring and / or the expression level of (b)CCNE1, and it is predicted that a human subject will respond to a CDK2 inhibitor if (1)(a) the presence of a CDKN2A gene encoding a p16 protein containing the amino acid sequence of SEQ ID NO: 1, (b) the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or (c) the presence of a p16 protein, and (2)(a) amplification of the CCNE1 gene and / or (b) an expression level of CCNE1 higher than the control expression level of CCNE1. In some embodiments, the subject has a CDK2-related disease or disorder. In some embodiments, the subject is suspected of having or at risk of developing a CDK2-related disease or disorder. In some embodiments, the method comprises (i) determining the presence of a CDKN2A gene lacking (a) the nucleotide sequence of the CDKN2A gene and / or (b) one or more inactivating nucleic acid substitutions and / or deletions from a biological sample obtained from a human subject, and (ii) determining the copy number of the CCNE1 gene from a biological sample obtained from a human subject, wherein the presence of (1)(a) the CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1 and / or (b) one or more inactivating nucleic acid substitutions and / or deletions from the CDKN2A gene, and (2)(a) the amplification of the CCNE1 gene are predicted to cause the human subject to respond to a CDK2 inhibitor.

[0009] In some embodiments of the aforementioned method, the amplification of the CCNE1 gene includes at least 3 gene copies. In some embodiments of the aforementioned method, the amplification of the CCNE1 gene includes at least 5 gene copies. In some embodiments of the aforementioned method, the amplification of the CCNE1 gene includes at least 21 gene copies.

[0010] In some embodiments of the method described above, the control expression level of CCNE1 is a predetermined cutoff value. In some embodiments of the method described above, the control expression level of CCNE1 is the expression level of CCNE1 in a sample(s) obtained from one or more subjects that did not respond to treatment with a CDK2 inhibitor.

[0011] In some embodiments of the aforementioned method, the CCNE1 expression level is the CCNE1 mRNA expression level. In some embodiments of the aforementioned method, the CCNE1 expression level is the CCNE1 protein expression level. In some embodiments where the CCNE1 expression level is the CCNE1 mRNA expression level, the CCNE1 expression level is measured by RNA sequencing, quantitative polymerase chain reaction (PCR), in situ hybridization, nucleic acid array, or RNA sequencing. In some embodiments where the CCNE1 expression level is the CCNE1 protein expression level, the CCNE1 expression level is measured by Western blotting, enzyme-linked immunosorbent, or immunohistochemical staining.

[0012] The disclosure also features a method for evaluating the CDKN2A and CCNE1 genes, the method comprising determining (i) the presence of (a) the nucleotide sequence of the CDKN2A gene or (b) the presence of one or more inactivated nucleic acid substitutions and / or deletions in the CDKN2A gene, and (ii) the copy number of the CCNE1 gene, from a biological sample(s) obtained from a human subject(s) having a disease or disorder associated with CDK2.

[0013] This disclosure also features a method for evaluating the response of a human subject to a CDK2 inhibitor in which the subject has, is suspected to have, or is at risk of developing a CDK2-related disease or disorder, the method comprising (a) administering a CDK2 inhibitor to a human subject, which has been previously determined to have amplification of the CCNE1 gene and / or a CCNE1 expression level higher than the control expression level of CCNE1, and (b) measuring the phosphorylation level of retinoblastoma (Rb) protein at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample obtained from the subject after administration in step (a), and comparing it to a control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, a decrease in the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 indicates that the human subject is responding to the CDK2 inhibitor. In some embodiments, the subject has a CDK2-related disease or disorder. In some embodiments, the subject has a CDK2-related disease or disorder, or is suspected to have, or is at risk of developing. In some embodiments, the biological sample includes a blood sample or a tumor biopsy sample.

[0014] This disclosure also features a method for measuring the amount of protein in a sample, the method comprising (a) providing a biological sample obtained from a human subject having a disease or disorder associated with CDK2, and (b) measuring the phosphorylation level of the Rb protein at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in the biological sample. In some embodiments, the biological sample includes a blood sample or a tumor biopsy sample.

[0015] In some embodiments of the method described above, the CDK2 inhibitor is one of the compounds described below, or a pharmaceutically acceptable salt thereof.

[0016] In some embodiments of the method described above, the disease or disorder associated with CDK2 is cancer.

[0017] Details of one or more embodiments of the present invention are shown in the accompanying drawings and the following description. Other features, purposes, and advantages of the present invention will become apparent from the description and drawings, and from the claims. [Brief explanation of the drawing]

[0018] [Figure 1] Characterization of ovarian and endometrial cell lines. A: The cell lines used in the study included four cell lines with CCNE1 amplification and three cell lines without CCNE1 amplification. CCNE1 amplification copy numbers are indicated. B: CCNE1 expression was determined by Western blotting in the indicated cell lines. This blot shows that cell lines with CCNE1 gain-of-function by copy number (CN>2) expressed higher levels of CCNE1 protein compared to cell lines with intermediate copy numbers or loss-of-function (CN≦2). GAPDH was detected as a loading control. Non-Amp indicates no amplification, Amp indicates amplification. [Figure 2A] siRNA-mediated CDK2 knockdown inhibits the proliferation of CCNE1-amplified cell lines. CCNE1-amplified Fu-ov1 (top) and KLE (bottom) cells were collected and subjected to cell cycle analysis 72 hours after transfection with either scrambled siRNA ("Ctl") or CDK2 siRNA. Cell cycle distribution was evaluated by FACS. Representative images from three separate experiments are shown. [Figure 2B] siRNA-mediated CDK2 knockdown inhibits the proliferation of CCNE1-amplified cell lines. CDK2 knockdown was confirmed by Western blotting after transfection with CDK2 siRNA. GAPDH was used as a loading control. [Figure 3A] CDK2 knockdown does not inhibit the proliferation of CCNE1 non-amplified strains. CCNE1 non-amplified COV504 and Igrov1 cells were collected and subjected to cell cycle analysis 72 hours after transfection with Ctl siRNA and CDK2 siRNA. Cell cycle distribution was evaluated by FACS. Representative images from three separate experiments are shown. [Figure 3B] CDK2 knockdown does not inhibit the proliferation of CCNE1 non-amp strains. CDK2 knockdown was confirmed by Western blotting after transfection with CDK2 siRNA. GAPDH was used as a loading control. [Figure 4] siRNA-mediated CDK2 knockdown inhibits proliferation in CCNE1-amplified human cancer cell lines but not in non-CCNE1-amplified human cancer cell lines. The percentage of S-phase cells 3 days after CDK2 siRNA transfection compared to Ctl siRNA is shown. Cell cycle distribution was evaluated by FACS. Mean values ​​represent three independent experiments in four CCNE1 amp cell lines and three non-amp cell lines. [Figure 5] Palbociclib treatment induces dose-dependent inhibition of proliferation in non-amplified CCNE1 cell lines, but not in amplified cell lines. Cell cycle analysis of non-amplified CCNE1 cell line COV504 (top) and CCNE1-amplified OVCAR3 cells (bottom) after 16 hours of palbociclib treatment. Cell cycle distribution was evaluated by FACS. [Figure 6] Palbociclib treatment selectively inhibits the proliferation of CCNE1 non-amplified cancer cell lines. The percentage of S-phase cells 16 hours after treatment with palbociclib at the indicated dose compared to DMSO. [Figure 7A] siRNA-mediated CDK2 knockdown blocks RB phosphorylation at S780 in CCNE1-amplified ovarian cells but not in non-amplified ovarian cells. Four CCNE1 amp cell lines, COV318, Fu-OV1, OVCAR3, and KLE cells, were transfected with CDK2 siRNA for 72 hours. Total protein was extracted from CDK2 siRNA or Ctl siRNA-transfected cells and subjected to Western blotting. GAPDH was used as a loading control. [Figure 7B]siRNA-mediated CDK2 knockdown blocks RB phosphorylation at S780 in CCNE1-amplified ovarian cells but not in non-amplified ovarian cells. Three CCNE1 non-amplified cell lines, COV504, OV56, and Igrov1, were transfected with CDK2 siRNA for 72 hours. Total protein was extracted from CDK2 siRNA or Ctl siRNA-transfected cells and subjected to Western blotting. GAPDH was used as a loading control. [Figure 8A] Palbociclib blocks RB phosphorylation at S780 in CCNE1 non-amplified ovarian cells but not in amplified ovarian cells. CCNE1 Amp OVCAR3 and COV318 cells were treated with various concentrations of palbociclib for 1 or 15 hours as shown. Total protein was extracted from these palbociclib or DMSO (control) treated cells and subjected to Western blotting. p-RB is phosphorylated retinoblastoma protein. GAPDH was used as a loading control. [Figure 8B] Palbociclib blocks RB phosphorylation at S780 in CCNE1 non-amplified ovarian cells but not in amplified ovarian cells. CCNE1 non-amplified COV504 and OV56 cells were treated with various concentrations of palbociclib for 1 or 15 hours as shown. Total protein was extracted from these palbociclib or DMSO (control) treated cells and subjected to Western blotting. p-RB is phosphorylated retinoblastoma protein. GAPDH was used as a loading control. [Figure 9]dTAG-mediated degradation of CDK2 reduces RB phosphorylation at S780. A: Chemical structure of dTAG. B: 14 hours of CDK2-dTAG treatment inhibited RB phosphorylation at S780 in CDK2 knockout OVCAR3 cells (right, Cas9+, CDK2-FKBP12(F36V)-HA+, CDK2-gRNA), but not in OVCAR3 cells with endogenous CDK2 (left, Cas9+, CDK2-FKBP12(F36V)-HA+, Ctl-gRNA). [Figure 10] p-RB S780 HTRF cell assay for identifying CDK2 inhibitors. A: IC50 in the CDK2 biochemical kinase activity assay. B: Concentration response analysis of the reference compound tested in the p-RB S780 HTRF cell assay. HTRF is a homogeneous time-resolved fluorescence assay. IC50 from the HTRF cell assay correlates with IC50 from the CDK2 enzyme assay. [Figure 11] Bioinformatics analysis of the CCLE dataset reveals that sensitivity to CDK2 inhibition in CCNE1-amplified cells is dependent on functional p16. The p16 status is shown in CDK2-sensitive versus insensitive cell lines. CCLE: Broad Institute Cancer Cell Line Encyclopedia (see Barretina below). [Figure 12] CCNE1-amplified cells with dysfunctional p16 do not respond to CDK2 inhibition. A: Western blot analysis of p16 in three gastric cell lines with CCNE1 amplification. B: Percentage of S-phase cells 3 days after CDK2 siRNA transfection compared to Ctl siRNA. Cell cycle distribution was evaluated by FACS. [Figure 13]siRNA-mediated p16 knockdown negates CDK2 inhibition-induced cell cycle repression in CCNE1-amplified cells. The percentage of S-phase cells after p16 knockdown and CDK2 inhibitor treatment was normalized to cells treated with Ctl siRNA and DMSO. CCNE1-amplified COV318 cells were transfected with either Ctl siRNA or p16 siRNA. 72 hours after transfection, cells were treated with 100 nM CDK2 inhibitor compound A. Cells were harvested and subjected to cell cycle analysis 16 hours after treatment. [Modes for carrying out the invention]

[0019] This disclosure provides predictive markers (e.g., biomarkers and pharmacodynamic markers, e.g., gene copy number, gene sequence, expression level, or phosphorylation level) for identifying human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder. This disclosure also provides pharmacodynamic markers (e.g., phosphorylation level) for identifying human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder that respond to CDK2 inhibitors. This disclosure also provides a method for treating human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder (e.g., cancer), the method comprising administering a CDK2 inhibitor to the human subjects.

[0020] CDK2-related diseases and disorders CDK2-related diseases or disorders are those in which the underlying pathology is mediated, either entirely or partially, by CDK2. Such diseases include cancer and other diseases involving proliferative disorders. In certain embodiments, CDK2-related diseases or disorders are treatable with CDK2 inhibitors.

[0021] In some embodiments, the disease or disorder associated with CDK2 is a cancerous tumor that includes an abnormality that activates CDK2 kinase activity. This includes, but is not limited to, cancers characterized by amplification or overexpression of CCNE1, such as ovarian cancer, uterine carcinosarcoma, and breast cancer, and cancers characterized by p27 inactivation, such as breast cancer and melanoma.

[0022] In some embodiments, the diseases or disorders associated with CDK2 are N-myc-amplified neuroblastoma (see Molenaar, et al., Proc Natl Acad Sci USA 106(31):12968-12973), K-Ras mutant lung cancer (see Hu, S., et al., Mol Cancer Ther, 2015.14(11):p.2576-85), or cancer with FBW7 mutation and CCNE1 overexpression (see Takada, et al., Cancer Res, 2017.77(18):p.4881-4893).

[0023] In some embodiments, the diseases or disorders associated with CDK2 are squamous cell carcinoma of the lung, adenocarcinoma of the lung, adenocarcinoma of the pancreas, invasive breast cancer, carcinosarcoma of the uterus, serous cystadenocarcinoma of the ovary, adenocarcinoma of the stomach, esophageal cancer, urothelial carcinoma of the bladder, mesothelioma, or sarcoma.

[0024] In some embodiments, the disease or disorder associated with CDK2 is lung adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, or gastric adenocarcinoma.

[0025] In some embodiments, the disease or disorder associated with CDK2 is adenocarcinoma, carcinoma, or cystadenoma.

[0026] In some embodiments, the disease or disorder associated with CDK2 is uterine cancer, ovarian cancer, gastric cancer, esophageal cancer, lung cancer, bladder cancer, pancreatic cancer, or breast cancer.

[0027] In some embodiments, the disease or disorder associated with CDK2 is cancer.

[0028] In some embodiments, the cancer is characterized by amplification or overexpression of CCNE1. In some embodiments, the cancer is ovarian cancer or breast cancer characterized by amplification or overexpression of CCNE1.

[0029] In some embodiments, breast cancer is resistant to chemotherapy or radiotherapy, endocrine-resistant, trastuzumab-resistant, or primary or acquired resistance to CDK4 / 6 inhibition. In some embodiments, breast cancer is advanced or metastatic breast cancer.

[0030] Examples of cancers treatable with CDK2 inhibitors using the methods of this disclosure include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, such as acute myeloid leukemia, chronic bone cancer. This includes, but is not limited to, myelin leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors in children, lymphocytic lymphoma, bladder cancer, kidney or urethral cancer, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers, including, for example, asbestos-induced cancer, Merkel cell carcinoma, and combinations of such cancers. The methods of this disclosure are also useful for treating metastatic cancers, particularly metastatic cancers expressing PD-L1.

[0031] In some embodiments, cancers treatable with CDK2 inhibitors using the methods of the present disclosure include melanoma (e.g., metastatic melanoma, BRAF, and HSP90 inhibitor-resistant melanoma), renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory adenocarcinoma of the prostate), breast cancer, colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), head and neck squamous cell carcinoma, urothelial carcinoma (e.g., bladder cancer), and high-frequency microsatellite instability (MSI). high Examples include cancers accompanied by ). Furthermore, this disclosure includes refractory or recurrent malignancies whose growth may be inhibited using the compounds of this disclosure.

[0032] In some embodiments, cancers treatable with CDK2 inhibitors using the methods of the present disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., lymphoma, leukemia, e.g., acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular lymphoma), Hodgkin lymphoma, or multiple myeloma, etc.), and combinations of such cancers.

[0033] In some embodiments, cancers treatable with a CDK2 inhibitor using the method of this disclosure include, but are not limited to, cholangiocarcinoma, bile duct cancer, triple-negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing's sarcoma, brain cancer, brain tumors, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, islet cell carcinoma, oral cancer, mouth cancer, pharyngeal cancer, laryngeal cancer, lip cancer, mesothelioma, cervical cancer, nasal cavity cancer, ocular cancer, melanoma of the eye, pelvic cancer, rectal cancer, renal cell carcinoma, salivary gland cancer, paranasal sinus cancer, spinal cancer, tongue cancer, tubular cancer, urethral cancer, and ureteral cancer.

[0034] In some embodiments, diseases and indications treatable with CDK2 inhibitors using the methods of this disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, urogenital tract cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

[0035] Exemplary blood cancers include lymphomas and leukemias, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular lymphoma), Hodgkin lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocytosis (ET)), myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), and multiple myeloma (MM).

[0036] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdosarcoma, fibroma, lipoma, hamartoma, and teratoma.

[0037] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), bronchogenic lung cancer, squamous cell carcinoma, anaplastic small cell carcinoma, anaplastic large cell carcinoma, adenocarcinoma, alveolar (bronchiolar) carcinoma, bronchial adenoma, chondrotoxic hamartoma, and mesothelioma.

[0038] Examples of gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine cancer (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma), and colorectal cancer.

[0039] Examples of urogenital tract cancers include cancers of the kidney (adenocarcinoma, Wilms' tumor [nephroblastoma]), cancers of the bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), cancers of the prostate (adenocarcinoma, sarcoma), and cancers of the testes (seminocarcinoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma).

[0040] Examples of liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0041] Examples of bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulosarcoma), multiple myeloma, malignant giant cell chordoma, osteochondroma (osteochondrial exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor.

[0042] Exemplary neurological cancers include cancers of the skull (osteoma, hemangioma, granuloma, xanthomas, osteoosteitis), cancers of the meninges (meningioma, meningiosarcoma, gliomas), cancers of the brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal glandoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), and cancers of the spinal cord (neurofibroma, meningioma, glioma, sarcoma), as well as neuroblastoma and Lhermitt-Dukuro disease.

[0043] Examples of gynecological cancers include cancer of the uterus (endometrial cancer), cancer of the cervix (cervical cancer, preneoplastic cervical dysplasia), cancer of the ovaries (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiable cancer), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), cancer of the vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), cancer of the vagina (clear cell carcinoma, squamous cell carcinoma, staphylosarcoma (embryonic rhabdomyosarcoma)), and cancer of the fallopian tubes (carcinoma).

[0044] Exemplary skin cancers include melanoma, basal cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, hemangioma, dermatofibroma, and keloid. In some embodiments, diseases and indications that can be treated with the compounds of this disclosure include, but are not limited to, triple-negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, cholangiocarcinoma, esophageal cancer, and urothelial carcinoma.

[0045] In some embodiments, the disease or disorder associated with CDK2 is an infection, such as a viral infection, bacterial infection, fungal infection, or parasitic infection.

[0046] Biomarkers and methods for predicting response to CDK2 inhibitors This specification provides biomarkers useful for predicting the responsiveness (e.g., improvement of disease status, as demonstrated by remission / recovery of disease) to CDK2 inhibitors in subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder. Accordingly, this specification also provides a method for predicting the response to CDK2 inhibitors in human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder. In certain embodiments, the prediction method described herein predicts that a subject will respond to treatment with a CDK2 inhibitor with an accuracy of at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or 100%. For example, in some embodiments, the prediction method described herein is applied to 10 subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, and 8 of those 10 subjects are predicted to respond to treatment with a CDK2 inhibitor based on the prediction method described herein, and 7 of those 8 subjects actually respond to treatment with a CDK2 inhibitor, then the accuracy of the prediction method is 87.5% (7 divided by 8). A subject is considered to respond to a CDK2 inhibitor if it shows any improvement in its disease state, such as a reduction or alleviation of symptoms, or remission / recovery of the disease.

[0047] CCNE1 and p16 In the examples, CCNE1 and p16 have been identified as genes that, when combined, are useful in predicting the responsiveness to CDK2 inhibitors (e.g., improvement of the disease as demonstrated by remission / recovery) in subjects with CDK2-related diseases or disorders.

[0048] p16 (also known as cyclin-dependent kinase inhibitor 2A, cyclin-dependent kinase 4 inhibitor A, multiple tumor suppressor 1, and p16-INK4a) functions as a negative regulator of normal cell proliferation by interacting with CDK4 and CDK6. p16 is encoded by the cyclin-dependent kinase inhibitor 2A ("CDKN2A") gene (GenBank accession number NM_000077). The cellular location of the CDKN2A gene is 9p21.3, which is position 21.3 on the short (p) arm of chromosome 9. The molecular location of the CDKN2A gene is base pairs 21,967,752–21,995,043 on chromosome 9 (Homo sapiens Annotation Release 109, GRCh38.p12). Genetic and epigenetic abnormalities in the gene encoding p16 are thought to lead to avoidance of aging and cancer development (Okamoto et al., 1994, PNAS 91(23):11045-9). Non-exclusive examples of genetic abnormalities in the gene encoding p16 are listed in Table 1 below. The amino acid sequence of human p16 is provided below (GenBank accession number NP_000068 / UniProtKB accession number P42771): 1 MEPAAGSSME PSADWLATAA ARGRVEEVRA LLEAGALPNA PNSYGRRPIQ VMMMGSARVA 61 ELLLLHGAEP NCADPATLTR PVHDAAREGF LDTLVVLHRA GARLDVRDAW GRLPVDLAEE 121 LGHRDVARYL RAAAGGTRGS NHARIDAAEG PSDIPD (Sequence ID 1).

[0049] CCNE1 is a cell cycle factor essential for regulating the G1 / S transition phase of the cell cycle (Ohtsubo et al., 1995, Mol.Cell.Biol.15:2612-2624). CCNE1 functions as a regulatory subunit of CDK2, interacting with CDK2 to form a serine / threonine kinase holoenzyme complex. The CCNE1 subunit of this holoenzyme complex provides substrate specificity for the complex (Honda et al., 2005, EMBO 24:452-463). CCNE1 is encoded by the cyclin E1 ("CCNE1") gene (GenBank accession number NM_001238). The amino acid sequence of human CCNE1 is provided below (GenBank accession number NP_001229 / UniProtKB accession number P24864): 1 mprerrerda kerdtmkedg gaefsarsrk rkanvtvflq dpdeemakid rtardqcgsq 61 pwdnnavcad pcsliptpdk edddrvypns tckpriiaps rgsplpvlsw anreevwkim 121 lnkektylrd qhfleqhpll qpkmrailld wlmevcevyk lhretfylaq dffdrymatq 181 envvktllql igisslfiaa kleeiyppkl hqfayvtdga csgdeiltme lmimkalkwr 241 lspltivswl nvymqvayln dlhevllpqy pqqifiqiae lldlcvldvd crefpygila 301 asalyhfsss elmqkvsgyq wcdiencvkw mvpfamvire tgssklkhfr gvadedahni 361 qthrdsldll dkarakkaml seqnrasplp sglltppqsg kkqssgpema (Sequence ID 2).

[0050] The examples demonstrate that CDK2 knockdown inhibits the proliferation of CCNE1-amplified cell lines, but not of CCNE1-non-amplified cell lines. Conversely, the examples show that CDK4 / 6 inhibition inhibits the proliferation of CCNE1-non-amplified cell lines, but not of CCNE1-amplified cell lines. The examples further demonstrate that the inhibition of cell proliferation observed in CCNE1-amplified cells treated with CDK2 inhibitors requires the presence of a normal (e.g., non-mutant or non-deletion) p16 gene. Therefore, CCNE1 and p16 are both combined biomarkers: that is, cells that respond to treatment with CDK2 inhibitors exhibit amplification of the CCNE1 gene and / or higher CCNE1 expression levels than the control expression level of CCNE1, possess a nucleotide sequence (e.g., gene or mRNA) encoding the p16 protein (e.g., the p16 protein containing the amino acid sequence of SEQ ID NO: 1), and / or the p16 protein is present, while control cells that do not respond to treatment with CDK2 inhibitors do not exhibit amplification of the CCNE1 gene and / or higher CCNE1 expression levels than the control expression level of CCNE1, possess a mutant or deletion gene encoding the p16 protein, and / or tend to lack p16 protein expression. Accordingly, the following methods are provided herein for using (i) amplification of the CCNE1 gene and / or the expression level of CCNE1, and (ii) the presence of a nucleotide sequence encoding the p16 protein, including the amino acid sequence of SEQ ID NO: 1, the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or the expression of the p16 protein, as biomarkers for predicting the response of a human subject to a CDK2 inhibitor in a human subject having, suspecting, or being at risk of having a CDK2-related disease or disorder. In certain embodiments, the human subject has a CDK2-related disease or disorder. In certain embodiments, the human subject is suspected of having, or is at risk of having, a CDK2-related disease or disorder.

[0051] In certain embodiments, a method is provided herein for predicting the response to a CDK2 inhibitor in human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, the method comprising (i) determining from a biological sample obtained from a human subject (a) the nucleotide sequence of the CDKN2A gene, (b) the presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) the presence of the p16 protein, and (ii) from a biological sample obtained from a human subject (a) CCN The process involves determining the copy number of the E1 gene and / or the expression level of (b)CCNE1, and it is predicted that a human subject will respond to a CDK2 inhibitor if (1)(a) the presence of a CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1, (b) the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or (c) the presence of the p16 protein, and (2)(a) amplification of the CCNE1 gene and / or (b) an expression level of CCNE1 higher than the control expression level of CCNE1. In certain embodiments, the human subject has a CDK2-related disease or disorder. In certain embodiments, the human subject is suspected of having or at risk of developing a CDK2-related disease or disorder. In certain embodiments, determining (i)(a) the nucleotide sequence of the CDKN2A gene, (b) the presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) the presence of the p16 protein is performed before administering the CDK2 inhibitor to a human subject (e.g., at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks, or 6 to 16 hours, 6 to 20 hours, or 6 to 24 hours, 2 to 3 days, 2 to 4 days, 2 to 5 days, 2 to 6 days, 2 to 7 days, 1 to 2 weeks, 1 to 3 weeks, or 1 to 4 weeks prior).In certain embodiments, (ii) determining (a) the copy number of the CCNE1 gene and / or (b) the expression level of CCNE1 in a biological sample obtained from a human subject is performed before administering the CDK2 inhibitor to the human subject (for example, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks, or 6 to 16 hours, 6 to 20 hours, or 6 to 24 hours, 2 to 3 days, 2 to 4 days, 2 to 5 days, 2 to 6 days, 2 to 7 days, 1 to 2 weeks, 1 to 3 weeks, or 1 to 4 weeks prior).

[0052] The presence of a CDKN2A gene encoding a p16 protein containing the amino acid sequence of SEQ ID NO: 1, the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or the presence of a p16 protein (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1), combined with amplification of the CCNE1 gene and / or a CCNE1 expression level higher than the control expression level of CCNE1, indicates / predicts that human subjects with, suspected of having, or at risk of developing, a CDK2-related disease or disorder will respond to CDK2 inhibitors.

[0053] In some embodiments, the CCNE1 gene is amplified to 3 to 25 copies. In certain embodiments, the CCNE1 gene is amplified to at least 3 copies. In certain embodiments, the CCNE1 gene is amplified to at least 5 copies. In certain embodiments, the CCNE1 gene is amplified to at least 7 copies. In certain embodiments, the CCNE1 gene is amplified to at least 10 copies. In certain embodiments, the CCNE1 gene is amplified to at least 12 copies. In certain embodiments, the CCNE1 gene is amplified to at least 14 copies. In certain embodiments, the CCNE1 gene is amplified to at least 21 copies.

[0054] In certain embodiments, the expression level of CCNE1 is the level of CCNE1 mRNA. In certain embodiments, the expression level of CCNE1 is the level of CCNE1 protein.

[0055] In certain embodiments, the CDKN2A gene encodes a protein containing the amino acid sequence of SEQ ID NO: 1.

[0056] In certain embodiments, one or more inactivating nucleic acid substitutions and / or deletions in the CDKN2A gene are as shown in Table 1. In certain embodiments, one or more inactivating nucleic acid substitutions and / or deletions in the CDKN2A gene are as described in Yarbrough et al., Journal of the National Cancer Institute, 91(18):1569-1574, 1999, Liggett and Sidransky, Biology of Neoplasia, Journal of Oncology, 16(3):1197-1206, 1998, and Cairns et al., Nature Genetics, 11:210-212, 1995 (each of which is incorporated herein by reference in its entirety). [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0057] Rb S780 Phosphorylation of Rb at serine (referred to herein as "Ser780" or "S780") corresponding to amino acid position 780 of Sequence ID No. 3 has been identified in examples as a useful pharmacodynamic marker for evaluating responsiveness to CDK2 inhibitors (e.g., CDK2 inhibition) in human subjects with diseases or disorders involving CCNE1 amplification.

[0058] Rb is a cell cycle regulator and functions as a tumor suppressor. Rb is activated by phosphorylation by cyclin D-CDK4 / 6 at Ser780 and Ser795, and by cyclin E / CDK2 at Ser807 and Ser811. Rb is encoded by the RB transcription corepressor 1 ("RB1") gene (GenBank accession number NM_000321). The amino acid sequence of human Rb is provided below (GenBank accession number NP_000312 / UniProtKB accession number P06400) (S780 is in bold and underlined): [ka]

[0059] As described above, the examples demonstrate that CDK2 knockdown inhibits the proliferation of CCNE1-amplified cell lines but not of CCNE1-non-amplified cell lines. The examples further demonstrate that CDK2 knockdown or inhibition blocks Rb phosphorylation at S780 in CCNE1-amplified cell lines but not in CCNE1-non-amplified cell lines. Therefore, Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is a pharmacodynamic marker for evaluating the response to CDK2 inhibition in CCNE1-amplified cancer cells or patients with diseases or disorders involving CCNE1 amplification. Accordingly, a method is provided herein relating to using the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 as a marker to predict the response of human subjects to CDK2 inhibitors in human subjects who have, are suspected of having, or are at risk of developing CDK2-related diseases or disorders, in which the human subjects have increased CCNE1 expression levels.

[0060] In certain embodiments, a method for evaluating the response to a CDK2 inhibitor in human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder is provided herein, and this method is (a) Human subjects have been administered a CDK2 inhibitor, and it has been determined that these human subjects have amplified CCNE1 genes and / or a higher CCNE1 expression level than the control expression level of CCNE1, and (b) The procedure involves measuring the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample obtained from a human subject after administration in step (a), A decrease in the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, compared to a control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, indicates that the human subject is responding to the CDK2 inhibitor. In certain embodiments, the human subject has a CDK2-related disease or disorder. In certain embodiments, the human subject is suspected of having, or at risk of developing, a CDK2-related disease or disorder. In certain embodiments, the human subject has further been determined to have a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions that prevent the CDKN2A gene from encoding a protein containing the amino acid sequence of SEQ ID NO: 1, and / or a p16 protein lacking one or more inactivating amino acid substitutions and / or deletions (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1). In certain embodiments, the measurement in step (b) is performed at least 6 hours, at least 16 hours, at least 20 hours, or at least 24 hours after the administration in step (a). In some embodiments, the measurement in step (b) is performed at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the administration in step (a). In certain embodiments, the measurement in step (b) is performed 6 to 16 hours, 6 to 20 hours, or 6 to 24 hours after the administration in step (a). In some embodiments, the measurement in step (b) is performed 2 to 3 days, 2 to 4 days, 2 to 5 days, 2 to 6 days, 2 to 7 days, 1 to 2 weeks, 1 to 3 weeks, or 1 to 4 weeks after the administration in step (a).

[0061] A decrease in Rb phosphorylation levels at serine corresponding to amino acid position 780 of SEQ ID NO: 3, when combined with CCNE1 gene amplification and / or CCNE1 expression levels higher than the control expression level, indicates that human subjects with, suspected of having, or at risk of developing, a CDK2-related disease or disorder respond to CDK2 inhibitors. For example, in subjects with CCNE1 gene amplification and / or CCNE1 expression levels higher than the control expression level, a low (e.g., decreased compared to control) or undetectable level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in biological samples obtained from the subjects after treatment with a CDK2 inhibitor indicates that the subjects respond to the CDK2 inhibitor.

[0062] When biological samples obtained from subjects after administration of a CDK2 inhibitor were compared to a control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, the decrease in Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, when combined with (i) amplification of the CCNE1 gene and / or a CCNE1 expression level higher than the control expression level of CCNE1, and (ii) the presence of a CDKN2A gene encoding a p16 protein containing the amino acid sequence of SEQ ID NO: 1, the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or the presence of a p16 protein (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1), indicates that human subjects with, suspected of having, or at risk of developing a CDK2-related disease or disorder respond to CDK2 inhibitors. For example, in a human subject having (i) amplification of the CCNE1 gene and / or a CCNE1 expression level higher than the control expression level of CCNE1, and (ii) the presence of a CDKN2A gene encoding a p16 protein containing the amino acid sequence of SEQ ID NO: 1, the presence of a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or the presence of a p16 protein (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1), a low (e.g., decreased compared to the control) or undetectable Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample obtained from the human subject after administration of a CDK2 inhibitor indicates that the human subject is responding to the CDK2 inhibitor.

[0063] In some embodiments, the CCNE1 gene is amplified to 3 to 25 copies. In certain embodiments, the CCNE1 gene is amplified to at least 3 copies. In certain embodiments, the CCNE1 gene is amplified to at least 5 copies. In certain embodiments, the CCNE1 gene is amplified to at least 7 copies. In certain embodiments, the CCNE1 gene is amplified to at least 10 copies. In certain embodiments, the CCNE1 gene is amplified to at least 12 copies. In certain embodiments, the CCNE1 gene is amplified to at least 14 copies. In certain embodiments, the CCNE1 gene is amplified to at least 21 copies.

[0064] In certain embodiments, the expression level of CCNE1 is the level of CCNE1 mRNA. In certain embodiments, the expression level of CCNE1 is the level of CCNE1 protein.

[0065] In certain embodiments, the CDKN2A gene encodes a protein containing the amino acid sequence of SEQ ID NO: 1.

[0066] In certain embodiments, one or more inactivating nucleic acid substitutions and / or deletions in the CDKN2A gene are as shown in Table 1. In certain embodiments, one or more inactivating nucleic acid substitutions and / or deletions in the CDKN2A gene are as described in Yarbrough et al., Journal of the National Cancer Institute, 91(18):1569-1574, 1999, Liggett and Sidransky, Biology of Neoplasia, Journal of Oncology, 16(3):1197-1206, 1998, and Cairns et al., Nature Genetics, 11:210-212, 1995 (each of which is incorporated herein by reference in its entirety).

[0067] contrast As described above, the method of the present invention may include measuring one or more markers (e.g., biomarkers or pharmacodynamic markers, e.g., amplification of the CCNE1 gene, CCNE1 expression level, presence of the CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1, presence of the CDKN2A gene with one or more inactivating nucleic acid substitutions and / or deletions, presence of the p16 protein (e.g., the p16 protein containing the amino acid sequence of SEQ ID NO: 1), and Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3) in a biological sample from a human subject who has, is suspected of having, or is at risk of developing, a disease or disorder associated with CDK2. In certain embodiments, the response of a human subject to a treatment including a CDK2 inhibitor is predicted / indicated by the level of one or more biomarkers (e.g., amplification (e.g., for the CCNE1 gene), expression level (e.g., for the CCNE1 or p16 protein), or phosphorylation level (e.g., for Rb)) compared to a control level of one or more biomarkers. In certain embodiments, a human subject is identified as potentially responsive to a CDK2 inhibitor if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than the control expression level of CCNE1, and (ii) a CDKN2A gene is present that encodes the p16 protein containing the amino acid sequence of SEQ ID NO: 1, a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions is present, and / or the p16 protein (e.g., the p16 protein containing the amino acid sequence of SEQ ID NO: 1) is present.In other embodiments, a human subject is identified as responding to a CDK2 inhibitor if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than the control expression level of CCNE1, and (ii) in a biological sample from a human subject after administration of a CDK2 inhibitor, the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is lower than the control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3. In yet another embodiment, a human subject is identified as responding to a CDK2 inhibitor if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than the control expression level of CCNE1; (ii) a CDKN2A gene encoding a p16 protein containing the amino acid sequence of SEQ ID NO: 1 is present, a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions is present, and / or a p16 protein (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1) is present; and (iii) in a biological sample from a human subject after administration of a CDK2 inhibitor, the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is lower than the control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3. In this context, the term “control” includes samples (from the same tissue type) obtained from human subjects known not to respond to a CDK2 inhibitor. The term “control” also includes samples (from the same tissue type) previously obtained from human subjects known not to respond to CDK2 inhibitors and used as a reference for future comparison with test samples taken from human subjects expected to be therapeutically responsive.A “control” level (e.g., gene copy number, expression level, or phosphorylation level) of a specific biomarker (e.g., CCNE1, p16, or Rb phosphorylation) in a particular cell type or tissue may be pre-defined by an analysis of biomarker levels (e.g., expression level or phosphorylation level) in one or more human subjects (e.g., two, three, four, five, six, seven, eight, nine, ten, fifteen, twenty, twenty-five, thirty, thirty-five, or forty or more) who did not respond to treatment with a CDK2 inhibitor. This pre-defined reference value (which may be the mean or median level (e.g., gene copy number, expression level, or phosphorylation level) obtained from multiple human subjects who did not respond to therapy) may then be used as the “control” level of the biomarker (e.g., CCNE1, p16, or Rb phosphorylation) in comparison to the test sample. In such comparisons, human subjects are predicted to respond to CDK2 inhibitors if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than a predefined reference, and if the CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1 is present, or if the CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions is present, and / or if the p16 protein (e.g., the p16 protein containing the amino acid sequence of SEQ ID NO: 1) is present. In another such comparison, human subjects are predicted to respond to CDK2 inhibitors if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than a predefined reference, and (ii) after administration of the CDK2 inhibitor to the human subjects, the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is lower than a predefined reference.In yet another such comparison, human subjects are shown to respond to CDK2 inhibitors if (i) the CCNE1 gene is amplified and / or the expression level of CCNE1 is higher than a pre-defined reference, (ii) a CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1 is present, a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions is present and / or the p16 protein (e.g., a p16 protein containing the amino acid sequence of SEQ ID NO: 1) is present, and (iii) after administration of the CDK2 inhibitor to human subjects, the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 is lower than a pre-defined reference.

[0068] Alternatively, a “control” level for a specific biomarker in a particular cell type or tissue may be predefined by an analysis of biomarker levels in one or more human subjects responding to treatment with a CDK2 inhibitor. This predefined reference value (which may be the mean or median level (e.g., expression level or phosphorylation level) obtained from multiple human subjects responding to the therapy) may then be used as the “control” level (e.g., expression level or phosphorylation level) in comparison with the test sample. In such a comparison, a human subject is indicated to respond to the CDK2 inhibitor if the level of the biomarker being analyzed (e.g., copy number of the CCNE1 gene, expression level of CCNE1, expression level of p16, or Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3) is equal to or equivalent to the predefined reference (e.g., at least 85% but less than 115% of the reference).

[0069] In certain embodiments, the “control” is a predetermined cutoff value. The cutoff value is typically a level of a biomarker (e.g., copy number, expression level, or phosphorylation level) that, when exceeded or below, is considered to predict the responsiveness of a human subject to the therapy of interest. Therefore, according to the methods and compositions described herein, a reference level (e.g., CCNE1 gene copy number, CCNE1 expression, p16 expression, or Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3) is identified as the cutoff value, and when exceeded or below, it predicts the responsiveness to the CDK2 inhibitor. The cutoff value determined for use in the methods described herein may, for example, be compared to a published concentration range, but may be individualized to the methodology and patient population used.

[0070] In some embodiments, the expression level of CCNE1 is increased compared to the expression level of CCNE1 in the control. For example, the expression level of CCNE1 analyzed is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 50 times, at least 75 times, or at least 100 times higher than the expression level of CCNE1 in the control, or at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, It could be at least 70%, at least 80%, at least 90%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 700%, at least 800%, at least 900%, at least 1,000%, at least 1,500%, at least 2,000%, at least 2,500%, at least 3,000%, at least 3,500%, at least 4,000%, at least 4,500%, or at least 5,000% higher.

[0071] The p16 protein is present if the protein is detectable by any assay known in the art or described herein, such as Western blotting, immunohistochemistry, fluorescence-activated cell sorting, and enzyme-linked immunoassays. In some embodiments, the p16 protein is present at an expression level of at least 5%, at least 10%, at least 20%, or at least 30% of the p16 expression level in healthy controls.

[0072] In some embodiments, the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 being analyzed is reduced compared to the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in the control. For example, the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 being analyzed is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 50 times, at least 75 times, or at least 100 times lower than the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in the control, or at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% lower.

[0073] Biological samples Suitable biological samples for the methods described herein include any sample containing blood or tumor cells obtained from or derived from a human subject requiring treatment. For example, a biological sample may contain tumor cells from a biopsy of a patient with a solid tumor. Tumor biopsies can be obtained by various means known in the art. Alternatively, a blood sample may be obtained from a patient with a blood cancer.

[0074] Biological samples may be obtained from human subjects who have, are suspected of having, or are at risk of developing a disease or disorder associated with CDK2. In some embodiments, the disease or disorder associated with CDK2 is cancer. In some embodiments, the disease or disorder associated with CDK2 is N-myc amplified neuroblastoma cells, K-Ras mutant lung cancer, and cancers with FBW7 mutation and CCNE1 overexpression. In some embodiments, the disease or disorder associated with CDK2 is lung squamous cell carcinoma, lung adenocarcinoma, pancreatic adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, gastric adenocarcinoma, esophageal cancer, bladder urothelial carcinoma, mesothelioma, or sarcoma. In some embodiments, the disease or disorder associated with CDK2 is lung adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, or gastric adenocarcinoma. In some embodiments, the disease or disorder associated with CDK2 is adenocarcinoma, carcinoma, or cystadenocarcinoma. In some embodiments, the disease or disorder associated with CDK2 is uterine cancer, ovarian cancer, gastric cancer, esophageal cancer, lung cancer, bladder cancer, pancreatic cancer, or breast cancer.

[0075] In some embodiments, breast cancer is resistant to chemotherapy or radiotherapy, endocrine-resistant, trastuzumab-resistant, or primary or acquired resistance to CDK4 / 6 inhibition. In some embodiments, breast cancer is advanced or metastatic breast cancer.

[0076] Methods for obtaining and / or preserving samples that preserve the activity or integrity of molecules (e.g., nucleic acids or proteins) within the sample are well known to those skilled in the art. For example, a biological sample may be further contacted with one or more additional agents, such as buffers and / or inhibitors, containing one or more nucleases, proteases, and phosphatase inhibitors, which preserve or minimize changes in the molecules within the sample.

[0077] Evaluation of biomarkers and pharmacodynamic markers The expression level of CCNE1 or p16 can be detected, for example, as RNA expression of the target gene (i.e., the gene encoding CCNE1 or p16). That is, the expression level (amount) of CCNE1 or p16 can be determined by detecting and / or measuring the level of mRNA expression of the gene encoding CCNE1. Alternatively, the expression level of CCNE1 or p16 can be detected, for example, as protein expression of the target gene (i.e., the gene encoding CCNE1 or p16). That is, the expression level (amount) of CCNE1 or p16 can be determined by detecting and / or measuring the level of protein expression of the gene encoding CCNE1 or p16.

[0078] In some embodiments, the expression level of CCNE1 or p16 is determined by measuring RNA levels. Various suitable methods can be used to detect and / or measure the level of mRNA expression of a gene. For example, mRNA expression can be determined using Northern blot or dot blot analysis, reverse transcriptase-PCR (RT-PCR, e.g., quantitative RT-PCR), in situ hybridization (e.g., quantitative in situ hybridization), nucleic acid arrays (e.g., oligonucleotide arrays or gene chips), and RNA sequencing analysis. Details of such methods are described below, as well as, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual Second Edition vol.1,2 and 3. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York, USA, Nov.1989; Gibson et al. (1999) Genome Res., 6(10):995-1001; and Zhang et al. (2005) Environ. Sci. Technol., 39(8):2777-2785; U.S. Patent Publication No. 2004086915; European Patent No. 0543942; and U.S. Patent No. 7,101,663; and Kukurba et al. (2015) Cold Spring Harbor Protocols., 2015(11):951-69 (each of which is incorporated herein by reference in its entirety).

[0079] For example, the presence or amount of one or more distinct mRNA populations in a biological sample can be determined by isolating the total mRNA from the biological sample (see, e.g., Sambrook et al. (above) and U.S. Patent No. 6,812,341), and separating the mRNA by size using agarose gel electrophoresis. The size-separated mRNA is then transcribed (e.g., by diffusion) onto a solid support such as a nitrocellulose membrane. The presence or amount of one or more mRNA populations in the biological sample can then be determined using one or more detectably labeled polynucleotide probes complementary to the mRNA sequence of interest, which bind to make their corresponding mRNA populations detectable. Detectable labels include, for example, fluorescent labels (e.g., umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansilchloride, allophycocyanin, or phycoerythrin), luminescence labels (e.g., europium, terbium, Qdot® nanoparticles supplied by Quantum Dot Corporation, Palo Alto, CA), radioactive labels (e.g., 125I, 131I, 35S, 32P, 33P, or 3H), and enzyme labels (horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase).

[0080] In some embodiments, the expression level of CCNE1 or p16 is determined by measuring protein levels. Various preferred methods can be used to detect and / or measure the level of protein expression of the target gene. For example, CCNE1 or p16 protein expression can be determined using Western blotting, enzyme-linked immunosorbent assay ("ELISA"), fluorescence-activated cell sorting, or immunohistochemical analysis (e.g., using CCNE1-specific or p16-specific antibodies, respectively). Details of such methods are described below and, for example, in Sambrook et al. above.

[0081] For example, the presence or amount of one or more distinct protein populations (e.g., CCNE1 or p16) in a biological sample can be determined by Western blotting, for example, by isolating the total protein from the biological sample (see, e.g., Sambrook et al. (above)), and separating the isolated protein by agarose gel electrophoresis to separate the proteins by size. The size-separated proteins are then transferred (e.g., by diffusion) to a solid support such as a nitrocellulose membrane. The presence or amount of one or more protein populations in the biological sample can then be determined using one or more antibody probes, for example, a first antibody specific to the protein of interest (e.g., CCNE1 or p16), and a second antibody that is detectably labeled and specific to the first antibody, which binds to make the corresponding protein population detectable. Detectable labels suitable for use in Western blotting are known in the art.

[0082] Methods for detecting or measuring gene expression (e.g., mRNA or protein expression) may be carried out in a form that allows for the rapid preparation, processing, and analysis of multiple samples. This may be done, for example, in a multi-well assay plate (e.g., 96-well or 386-well) or array (e.g., nucleic acid chip or protein chip). Stock solutions of various reagents may be provided manually or under robotic control, and subsequent sample preparation (e.g., RT-PCR, labeling, or cell fixation), pipetting, dilution, mixing, distribution, washing, incubation (e.g., hybridization), sample reading, data acquisition (optical data), and / or analysis (computer-aided image analysis) may be performed under robotic control using commercially available analysis software, robotics, and detection instruments capable of detecting signals generated from assays. Examples of such detectors include, but are not limited to, spectrophotometers, luminometers, fluorometers, and instruments for measuring radioisotope decay. Exemplary high-throughput cell-based assays (e.g., detecting the presence or level of a target protein within a cell) can utilize ArrayScan® VTI HCS reader or KineticScan® HCS reader technology (Cellomics Inc., Pittsburg, PA).

[0083] In some embodiments, the presence of a CDKN2A gene encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1 and / or a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions is determined by evaluating the DNA sequence of the CDKN2A gene (e.g., genomic DNA or cDNA) or by evaluating the RNA sequence of the CDKN2A gene (e.g., RNA, e.g., mRNA). Methods for performing nucleic acid sequencing analysis are known in the art and are described above. Non-limiting examples of inactivating nucleic acid substitutions and / or deletions that prevent the CDKN2A gene from encoding a protein containing the amino acid sequence of SEQ ID NO: 1 are listed in Table 1 above. In certain embodiments, one or more inactivating nucleic acid substitutions and / or deletions in the CDKN2A gene are as described in Yarbrough et al., Journal of the National Cancer Institute, 91(18):1569-1574, 1999, Liggett and Sidransky, Biology of Neoplasia, Journal of Oncology, 16(3):1197-1206, 1998, and Cairns et al., Nature Genetics, 11:210-212, 1995 (each of which is incorporated herein by reference in its entirety).

[0084] In some embodiments, the expression level or presence of a gene lacking one or more inactivating nucleic acid substitutions or deletions is determined by evaluating the copy number variation (CNV) of the gene. The CNV of a gene (e.g., the CCNE1 gene and / or the CDKN2A gene) can be determined / identified by a variety of preferred methods. For example, CNVs can be determined using fluorescence in situ hybridization (FISH), multiplex ligation-dependent probe amplification (MLPA), array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) arrays, and next-generation sequencing (NGS) techniques.

[0085] For example, copy number variations of one or more distinct genes in a biological sample can be determined by MLPA by, for instance, extracting a DNA sample from the biological sample (see, e.g., Sambrook et al. (above) and U.S. Patent No. 6,812,341) and amplifying the target DNA sequence (e.g., CCNE1 or CDKN2A) using a mixture of MLPA probes. Each MLPA probe consists of two oligonucleotides that hybridize to immediately adjacent target DNA sequences (e.g., CCNE1 or CDKN2A) to ligate into a single probe. The ligated probe is amplified by PCR using a single fluorescently labeled PCR primer, allowing visualization of the amplified product during fragment separation by capillary electrophoresis. The presence, absence, or amplification of one or more target genes in the biological sample is calculated by measuring the fluorescence derived from PCR, quantifying the amount of normalized PCR product, and comparing it to a control DNA sample.

[0086] The level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 can be detected by various preferred methods. For example, the phosphorylation state can be determined using Western blotting, ELISA, fluorescence-activated cell sorting, or immunohistochemical analysis. Details of such methods are described below and, for example, by Sambrook et al. above.

[0087] Similar to the methods for detecting or measuring gene expression (described above), a method for detecting or measuring the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 may be provided in a form that allows for the rapid preparation, processing, and analysis of multiple samples.

[0088] CDK2 inhibitors Compounds useful in the methods of this disclosure are CDK2 inhibitors. In some embodiments, the CDK2 inhibitor inhibits CDK2, CDK4, and CDK6. In some embodiments, the CDK2 inhibitor selectively inhibits CDK2 more than CDK1 and CDK9. In some embodiments, the CDK2 inhibitor selectively inhibits CDK2 more than CDK4 and CDK6. In some embodiments, the CDK2 inhibitor selectively inhibits CDK2 more than CDK1, CDK9, CDK4, and CDK6. In some embodiments, the compound is ICed according to the methods of Examples A, B, and C. 50 As can be calculated by measuring, the compounds are about 2, 3, 5, 10, 15, or 20 times more selective to CDK2 than to CDK1 and CDK9. In some embodiments, the compounds are IC2-100 according to the methods of Examples A, B, C, D, and E. 50 As can be calculated by measuring, the compounds are about 2 times, 3 times, 5 times, 10 times, 15 times, or 20 times more selective than CDK2 than CDK1, CDK9, CDK4, or CDK6. In some embodiments, the compounds are IC2-100 according to the methods of Examples A, D, and E. 50 As can be calculated by measuring, it is approximately 2, 3, 5, 10, 15, or 20 times more selective than CDK2 than CDK4 and CDK6.

[0089] In some embodiments, the CDK2 inhibitor is dinaciclib (Merck), alvociclib (Tolero Pharmaceuticals), cericclib (Cyclacel Pharmaceuticals), ronicicclib (Bayer), milciclib (Nerviano), abemacicclib (Eli Lilly), trilacricib (G1 Therapeutics), CYC065 (Cyclacel Pharmaceuticals), AT-7519 (Astex Therapeutics, J Med.Chem., 2008, 51, 4986), BMS-387032 / SNS032 (Sunesis, J Med.Chem., 2004, 47, 1719), TG02 (Trajara The pharmaceutically acceptable salts of the following: R547 (Roche, Mol. Can. Ther. 2006, 2644), AZD5438 (AstraZeneca, Mol. Can. Ther. 2009, 1856), RGB-286638 (Agennix, Leukemia, 2013, 2366), AMG295 (Amgen, WO2009 / 085185), PHA-793887 (Nerviano, BMC, 2010 18, 1844), ZK-304709 (Biomed. Pharmacother. 2006, 269), and AG-024322 (Pfizer, Cancer Res. 2005, 1045), or any one of the aforementioned. The chemical structures of CYC065, AT7519, BMS-387032 / SN032, TG02, R547, AZD5438, RGB-286638, AMG925, PHA-793887, ZK-304709, and AG-24322 are provided below: [ka] [ka]

[0090] In some embodiments, the CDK2 inhibitor is compound A(8-((1R,2R)-2-hydroxy-2-methylcyclopentyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrido[2,3-d]pyrimidine-7(8H)-one), having the following structure, or a pharmaceutically acceptable salt thereof: [ka] (8-[(1R,2R)-2-hydroxy-2-methylcyclopentyl]-2-{[1-(methylsulfonyl)piperidine-4-yl]amino}pyrido[2,3-d]pyrimidine-7(8H)-one, see page 51, paragraph

[0987] of U.S. Patent Application Publication No. 2018 / 0044344, which is incorporated herein by reference in its entirety).

[0091] In some embodiments, the compound is a compound in either an embodiment or an exemplary compound in U.S. Patent Application Publication No. 2018 / 0044344 (which is incorporated herein by reference in its entirety), or a pharmaceutically acceptable salt thereof.

[0092] In some embodiments, the compound is a compound in any embodiment or exemplary compound in U.S. Patent Application No. 16 / 598,777 filed October 10, 2019, or in U.S. Provisional Patent Application No. 62 / 806,269 filed February 15, 2019 (each of which is incorporated herein by reference in its entirety), or a pharmaceutically acceptable salt thereof.

[0093] In certain embodiments, the CDK2 inhibitor is given by formula (AI): [ka] A compound of or a pharmaceutically acceptable salt thereof, in which, R 1 However, H, C 1~6 Alkyl and C 1~6 Selected from haloalkyl groups, R 2 However, C1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~10 cycloalkyl, C 6~10 aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, C 6~10 aryl-C 1~4 alkyl, 4- to 10-membered heterocycloalkyl-C 1~4 alkyl, 5- to 10-membered heteroaryl-C 1~4 alkyl, C(=O)R b , C(=O)NR c R d , C(=O)OR a , C(=NR e )R b , C(=NR e )NR c R d , S(=O)R b , S(=O)NR c R d , NR c S(=O)2R b , NR c S(=O)2NR c R d , S(=O)2R b , and S(=O)2NR c R d selected from, and the C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~10 cycloalkyl, C 6~10 aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, C 6~10 aryl-C 1~4 alkyl, 4- to 10-membered heterocycloalkyl-C 1~4 alkyl, and 5- to 10-membered heteroaryl-C 1~4 Each alkyl is one, two, three, or four independently selected R 2Aoptionally substituted with a substituent, each R a 、R c 、and R d is independently selected from H, C 1~6 alkyl, C 1~6 haloalkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 3~10 cycloalkyl, C 6~10 aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, C 6~10 aryl-C 1~4 alkyl, 4- to 10-membered heterocycloalkyl-C 1~4 alkyl, and 5- to 10-membered heteroaryl-C 1~4 alkyl, and the C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~10 cycloalkyl, C 6~10 aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, C 6~10 aryl-C 1~4 alkyl, 4- to 10-membered heterocycloalkyl-C 1~4 alkyl, and 5- to 10-membered heteroaryl-C 1~4 alkyls are each optionally substituted with one, two, three, or four independently selected R 2A substituents, each R b is C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~10 cycloalkyl, C 6~10 aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, C 6~10 aryl-C 1~4Alkyl, 4- to 10-member heterocycloalkyl-C 1~4 Alkyl, and 5- to 10-member heteroaryl-C 1~4 Independently selected from alkyl, each of which is optionally substituted with one, two, three, or four independently selected R 2A Substituents, Each R e Is H, CN, OH, C 1~4 Alkyl, and C 1~4 Independently selected from alkoxy, Each R f Is H, C 1~4 Alkyl, and C 1~4 Independently selected from haloalkyl, R 3 Is C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4- to 10-member heterocycloalkyl, 5- to 10-member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4- to 10-member heterocycloalkyl-C 1~4 Alkyl, and 5- to 10-member heteroaryl-C 1~4 Selected from alkyl, each of which is optionally substituted with one, two, three, or four independently selected R 3A Substituents,[[ID=5​​​​​​​​​​​​​​​​​​​​​​​​​​​​Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Each cycloalkyl group can be one, two, three, or four independently selected R groups. G Substituents may be substituted, Or, instead, R 4 and R 5 However, together with the carbon atoms to which they are bonded, they form a 3, 4, 5, 6, or 7-membered cycloalkyl ring or a 3, 4, 5, 6, or 7-membered heterocycloalkyl ring, each of which has one, two, three, or four independently selected R G Substituents may be used, R 6 and R 7 However, H, D, Haro, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Selected independently from cycloalkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Each cycloalkyl group can be one, two, three, or four independently selected R groups. G Substituents may be substituted, Or, instead, R 6 and R 7 However, together with the carbon atoms to which they are bonded, they form a 3, 4, 5, 6, or 7-membered cycloalkyl ring or a 3, 4, 5, 6, or 7-membered heterocycloalkyl ring, each of which has one, two, three, or four independently selected R G Substituents may be used, Group (B): R 4 and R 5 However, H, Haro, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C3~6 Selected independently from cycloalkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Each cycloalkyl group can be one, two, three, or four independently selected R groups. G Substituents may be substituted, Or, instead, R 4 and R 5 However, together with the carbon atoms to which they are bonded, they form a 3, 4, 5, 6, or 7-membered cycloalkyl ring or a 3, 4, 5, 6, or 7-membered heterocycloalkyl ring, each of which has one, two, three, or four independently selected R G Substituents may be used, R 6 and R 7 But, hello, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Selected independently from cycloalkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl and C 3~6 Each cycloalkyl group can be one, two, three, or four independently selected R groups. G Substituents may be substituted, Or, instead, R 6 and R 7 However, together with the carbon atoms to which they are bonded, they form a 3, 4, 5, 6, or 7-membered cycloalkyl ring or a 3, 4, 5, 6, or 7-membered heterocycloalkyl ring, each of which has one, two, three, or four independently selected R G Substituents may be used, Each R 2A However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl, 5-10 member heteroaryl-C 1~4 Alkyl, OR a1 , SR a1 , C(=O)R b1 , C(=O)NR c1 R d1 , C(=O)OR a1 ,OC(=O)R b1 , OC(=O)NR c1 R d1 , NR c1 R d1 , NR c1 C(=O)R b1 , NR c1 C(=O)OR b1 , NR c1 C(=O)NR c1 R d1 , C(=NR e )R b1 , C(=NR e )NR c1 R d1 , NR c1 C(=NR e )NR c1 R d1 NHOR a1 , NR c1 S(=O)R b1 , NR c1 S(=O)NR c1 R d1 , S(=O)R b1 , S(=O)NR c1 R d1 , NR c1 S(=O)2R b1 , NR c1 S(=O)2NR c1 R d1 , S(=O)2R b1 , S(=O)(=NR f )R b1 , and S(=O)2NRc1 R d1 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 2B Substituents may be used, Each R a1 , R c1 , and R d1 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 2B Substituents may be used, Each R b1 However, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 2B Substituents may be used, Each R 3A However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl, 5-10 member heteroaryl-C 1~4 Alkyl, OR a2 , SR a2 , C(=O)R b2 , C(=O)NR c2 R d2 , C(=O)OR a2 ,OC(=O)R b2, OC(=O)NR c2 R d2 , NR c2 R d2 , NR c2 C(=O)R b2 , NR c2 C(=O)OR b2 , NR c2 C(=O)NR c2 R d2 , C(=NR e )R b2 , C(=NR e )NR c2 R d2 , NR c2 C(=NR e )NR c2 R d2 NHOR a2 , NR c2 S(=O)R b2 , NR c2 S(=O)NR c2 R d2 , S(=O)R b2 , S(=O)NR c2 R d2 , NR c2 S(=O)2R b2 , NR c2 S(=O)2NR c2 R d2 , S(=O)2R b2 , S(=O)(=NR f )R b2 , and S(=O)2NR c2 R d2 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4Each alkyl group can be one, two, three, or four independently selected R 3B Substituents may be used, Each R a2 , R c2 , and R d2 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 3B Substituents may be used, Each R b2 However, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, C 6~10 Aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4Alkyl, C 6~10 Aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 3B Substituents may be used, Each R 2B and R 3B However, H, D, Haro, CN, NO2, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a23 , SR a23 , C(=O)R b23 , C(=O)NR c23 R d23 , C(=O)OR a23 ,OC(=O)R b23 , OC(=O)NR c23 R d23 , NR c23 R d23 , NR c23 C(=O)R b23 , NR c23 C(=O)OR b23 , NR c23 C(=O)NR c23 R d23 , C(=NR e )R b23 , C(=NR e )NR c23 R d23 , NR c23 C(=NR e )NR c23 R d23 NHOR a23 , NR c23 S(=O)R b23 , NR c23 S(=O)NR c23 R d23 , S(=O)Rb23 , S(=O)NR c23 R d23 , NR c23 S(=O)2R b23 , NR c23 S(=O)2NR c23 R d23 , S(=O)2R b23 , S(=O)(=NR f )R b23 , and S(=O)2NR c23 R d23 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R G Substituents may be used, Each R a23 , R c23 , and R d23 However, H, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R G Substituents may be used, Each R b23 However, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. G Substituents may be used, Each R G However, OH, NO2, CN, Halo, C 1~3 Alkyl, C 2~3 Alkenil, C 2~3 Alkinyl, C 1~3 Haloalkyl, cyano-C 1~3 Alkyl, HO-C 1~3 Alkyl, C 1~3 Alkoxy-C 1~3 Alkyl, C 1~3 Alkoxy, C 1~3 Haloalkoxy, amino, C 1~3 Alkylamino, di(C 1~3 Alkyl)amino, thio, C 1~3 Alkylthio, C 1~3 Alkyl sulfinyl, C 1~3 Alkyl sulfonyl, carbamyl, C 1~3 Alkylcarbamyl, di(C1~3 Alkyl)carbamyl, carboxy, C 1~3 Alkylcarbonyl, C 1~3 Alkoxycarbonyl, C 1~3 Alkylcarbonyloxy, C 1~3 Alkylcarbonylamino, C 1~3 Alkoxycarbonylamino, C 1~3 Alkylaminocarbonyloxy, C 1~3 Alkylsulfonylamino, aminosulfonyl, C 1~3 Alkylaminosulfonyl, di(C 1~3 Alkyl)aminosulfonyl, aminosulfonylamino, C 1~3 Alkylaminosulfonylamino, di(C 1~3 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1~3 Alkylaminocarbonylamino, and di(C 1~3 It is independently selected from alkyl)aminocarbonylamino.

[0094] In some embodiments, R 1 H is H.

[0095] In some embodiments, R 2 C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected by one, two, three, or four R groups. 2A Substitutions may be used.

[0096] In some embodiments, R 2 R is selected from 4- to 7-membered heterocycloalkyls and phenyls, each of which is selected from 1, 2, 3, or 4 independently. 2A It is substituted with a substituent.

[0097] In some embodiments, R 2 It is selected from piperidine-4-yl and phenyl, each of which has one R 2A Substitutions may be used.

[0098] In some embodiments, at least one R 2A However, S(=O)2R b1 and S(=O)2NR c1 R d1 Selected from, R b1 is C 1~3 It is alkyl, R c1 and R d1 H and C 1~3 It is selected independently of alkyl.

[0099] Several embodiments, each R 2A This is selected independently from S(=O)2CH3 and S(=O)2NH2.

[0100] In some embodiments, R 2 is S(=O)2R b1 It is either piperidine-4-yl substituted with R 2 is S(=O)2NR c1 R d1 It is a phenyl compound substituted with [a specific compound].

[0101] In some embodiments, R 2 is either piperidine-4-yl substituted with S(=O)2CH3, or R 2 This is phenyl substituted with S(=O)2NH2.

[0102] In some embodiments, R 3 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected by one, two, three, or four R groups. 3A Substitutions may be used.

[0103] In some embodiments, R 3 C 1~6 Alkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is one or two independently selected R 3A Substitutions may be used.

[0104] In some embodiments, one, two, three, or four independently selected R 3A R which may be substituted by substituents 3 The compound is selected from 1,1-difluorobutan-2-yl, cyclopentyl, phenyl, tetrahydrofuran-3-yl, and (1-methyl-1H-pyrazole-5-yl)methyl.

[0105] Several embodiments, each R 3A H, Halo, C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0106] In some embodiments, R 4 and R 5 Each is C 1~6 Alkyl and C 1~6 Independently selected from haloalkyls, or instead, R 4 and R 5 These atoms, together with the carbon atoms to which they are bonded, form a 3, 4, 5, or 6-membered cycloalkyl ring.

[0107] In some embodiments, R 4and R 5 These, together with the carbon atoms to which they are bonded, form a 3, 4, 5, 6, or 7-membered cycloalkyl ring.

[0108] In some embodiments, R 4 and R 5 These, along with the carbon atoms to which they are bonded, form a cyclopropyl ring.

[0109] In some embodiments, R 4 and R 5 C 1~3 Alkyl or C 1~3 It is a haloalkyl group.

[0110] In some embodiments, R 4 and R 5 C 1~3 It is alkyl.

[0111] In some embodiments, R 4 and R 5 It is independently methyl.

[0112] In some embodiments, R 4 and R 5 They form a cyclopropyl ring with the carbon atoms to which they are bonded, or R 4 and R 5 C 1~3 It is alkyl.

[0113] In some embodiments, R 6 and R 7 H and C are respectively 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0114] In some embodiments, R 6 and R 7 Each of these is H.

[0115] In some embodiments, R 1 H is H, R 2 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected by one, two, three, or four R groups. 2A Substituents may be used, R 3 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected by one, two, three, or four R groups. 3A Substituents may be used, R 4 and R 5 Each is C 1~6 Alkyl and C 1~6 Independently selected from haloalkyl groups, Or, instead, R 4 and R 5 These, together with the carbon atoms to which they are bonded, form a 3, 4, 5, or 6-membered cycloalkyl ring. R 6 and R 7 H and C 1~6 Selected independently of alkyl, Each R 2A Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, OR a1 , SR a1 , C(=O)R b1 , C(=O)NR c1 R d1 , C(=O)OR a1 ,OC(=O)R b1 , OC(=O)NR c1 R d1 , NR c1 R d1 , NR c1 C(=O)R b1 , NR c1 C(=O)OR b1 , NR c1 C(=O)NR c1 R d1 NHOR a1 , NR c1 S(=O)2R b1 , NR c1 S(=O)2NR c1 R d1 , S(=O)2R b1 , and S(=O)2NR c1 R d1 Selected independently from, Each R a1 , R c1 , and R d1 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R b1 C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R 3A Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, OR a2 , SR a2 , C(=O)R b2 , C(=O)NR c2 R d2, C(=O)OR a2 ,OC(=O)R b2 , OC(=O)NR c2 R d2 , NR c2 R d2 , NR c2 C(=O)R b2 , NR c2 C(=O)OR b2 , NR c2 C(=O)NR c2 R d2 NHOR a2 , NR c2 S(=O)2R b2 , NR c2 S(=O)2NR c2 R d2 , S(=O)2R b2 , and S(=O)2NR c2 R d2 Selected independently from, Each R a2 , R c2 , and R d2 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R b2 C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0116] In some embodiments, R 1 H is H, R 2 These are selected from 4- to 7-membered heterocycloalkyl groups and phenyl groups, each having one R 2A Substituted by the group, R 2A is S(=O)2R b1 Or S(=O)2NR c1 R d1 And, R b1 is C 1~3 It is alkyl, R c1 and R d1 H and C 1~3 Selected independently of alkyl, R 3 C 1~6 Alkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected by one, two, three, or four R groups. 3A Substituents may be used, Each R 3A H, Halo, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, R 4 and R 5 Each of them is methyl, or R 4 and R 5 They form a cyclopropyl ring together with the carbon atoms to which they are bonded. R 6 and R 7 Each of these is H.

[0117] In certain embodiments, the CDK2 inhibitor is given by formula (BI): [ka] A compound of or a pharmaceutically acceptable salt thereof, in which, n is an integer selected from 0, 1, 2, 3, 4, 5, and 6. Ring portion A is a 3-14 member cycloalkyl or a 4-14 member heterocycloalkyl, and ring portion A is bonded to the NH group of formula (I) on a saturated or partially saturated ring of the 3-14 member cycloalkyl or 4-14 member heterocycloalkyl, R 1 However, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~14 Cycloalkyl, 6-14 membered aryl, 4-14 membered heterocycloalkyl, 5-14 membered heteroaryl, C 3~14 Cycloalkyl-C 1~4 Alkyl, 6-14 member aryl-C1~4 Alkyl, 4-14 member heterocycloalkyl-C 1~4 Alkyl and 5-14 member heteroaryl-C 1~4 Selected from alkyl groups, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~14 Cycloalkyl, 6-14 membered aryl, 4-14 membered heterocycloalkyl, 5-14 membered heteroaryl, C 3~14 Cycloalkyl-C 1~4 Alkyl, 6-14 member aryl-C 1~4 Alkyl, 4-14 member heterocycloalkyl-C 1~4 Alkyl and 5-14 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, four, five, or six independently selected R 4 Substitutable by substituents, R 2 and R 3 Each of them is C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 A C is independently selected from cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl. 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl are each selected in one, two, three, or four independent R groups. G Substituents may be used to replace, or R 2 and R 3 However, together with the carbon atoms to which they are bonded, they form ring B. Ring B is a 3- to 7-membered cycloalkyl ring or a 4- to 7-membered heterocycloalkyl ring, each of which has one, two, three, or four independently selected R groups. GSubstitutable by substituents, Each R 4 However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl, 5-10 member heteroaryl-C 1~4 Alkyl, OR a4 , SR a4 NHOR a4 , C(O)R b4 , C(O)NR c4 R d4 , C(O)NR c4 (OR a4 ), C(O)OR a4 ,OC(O)R b4 , OC(O)NR c4 R d4 , NR c4 R d4 , NR c4 NR c4 R d4 , NR c4 C(O)R b4 , NR c4 C(O)OR a4 , NR c4 C(O)NR c4 R d4 , C(=NR e4 )R b4 , C(=NR e4 )NR c4 R d4 , NR c4 C(=NR e4 )NR c4 R d4 , NR c4 C(=NR e4 )R b4 , NR c4 S(O)NR c4 R d4 , NRc4 S(O)R b4 , NR c4 S(O)2R b4 , NR c4 S(O)(=NR e4 )R b4 , NR c4 S(O)2NR c4 R d4 S(O)R b4 , S(O)NR c4 R d4 S(O)2R b4 , S(O)2NR c4 R d4 OS(O)(=NR e4 )R b4 OS(O)2R b4 , S(O)(=NR e4 )R b4 SF5, P(O)R f4 R g4 , OP(O)(OR h4 )(OR i4 ), P(O)(OR h4 )(OR i4 ), and BR j4 R k4 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 4A Substituents may be used, Each R 5 However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C3~10 Cycloalkyl, 6- to 10-membered aryl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6- to 10-membered aryl-C 1~4 Alkyl, 4- to 10-membered heterocycloalkyl-C 1~4 Alkyl, 5- to 10-membered heteroaryl-C 1~4 Alkyl, OR a5 , SR a5 , NHOR a5 , C(O)R b5 , C(O)NR c5 R d5 , C(O)NR c5 (OR a5 ), C(O)OR a5 , OC(O)R b5 , OC(O)NR[[ID=3,2]] c5 R d5 , NR c5 R d5 , NR c5 NR c5 R d5 , NR c5 , C(O)R b5 , NR c5 , C(O)OR a5 , NR c5 , C(O)NR c5 R d5 , C(=NR e5 )R b5 , C(=NR e5 )NR c5 R d5 , NR c5 , C(=NR e5 )NR c5 R d5 , NR c5 , C(=NR e5 )R b5 , NR c5 , S(O)NR c5 R d5 , NR c5 , S(O)R b5 , NR c5 , S(O)2R b5 , NR c5 , S(O)(=NR e5 )R b5 , NR c5 , S(O)2NRc5 R d5 、 S(O)R b5 、 S(O)NR c5 R d5 、 S(O)₂R b5 、 S(O)₂NR c5 R d5 、 OS(O)(=NR e5 )R b5 、 OS(O)₂R b5 、 S(O)(=NR e5 )R b5 、 SF₅、 P(O)R f5 R g5 、 OP(O)(OR h5 )(OR i5 )、 P(O)(OR h5 )(OR i5 )、 and BR j5 R k5 is independently selected from, and the C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~10 cycloalkyl, 6-10 member aryl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 cycloalkyl-C 1~4 alkyl, 6-10 member aryl-C 1~4 alkyl, 4-10 member heterocycloalkyl-C 1~4 alkyl, and 5-10 member heteroaryl-C 1~4 alkyl are each optionally substituted with one, two, three, or four independently selected R 5A substituents, each R 4A is H, D, halo, CN, NO₂, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, C 1~6 haloalkyl, C 3~7 cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 cycloalkyl-C 1~4 alkyl, phenyl-C 1~4 alkyl, 4-7 member heterocycloalkyl-C 1~4Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a41 , SR a41 NHOR a41 , C(O)R b41 , C(O)NR c41 R d41 , C(O)NR c41 (OR a41 ), C(O)OR a41 ,OC(O)R b41 , OC(O)NR c41 R d41 , NR c41 R d41 , NR c41 NR c41 R d41 , NR c41 C(O)R b41 , NR c41 C(O)OR a41 , NR c41 C(O)NR c41 R d41 , C(=NR e41 )R b41 , C(=NR e41 )NR c41 R d41 , NR c41 C(=NR e41 )NR c41 R d41 , NR c41 C(=NR e41 )R b41 , NR c41 S(O)NR c41 R d41 , NR c41 S(O)R b41 , NR c41 S(O)2R b41 , NR c41 S(O)(=NR e41 )R b41 , NR c41 S(O)2NR c41 R d41 S(O)R b41 , S(O)NR c41 R d41 S(O)2R b41 , S(O)2NR c41 R d41 OS(O)(=NR e41 )Rb41 OS(O)2R b41 , S(O)(=NR e41 )R b41 SF5, P(O)R f41 R g41 , OP(O)(OR h41 )(OR i41 ), P(O)(OR h41 )(OR i41 ), and BR j41 R k41 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 4B Substituents may be used, Each R 4B However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a42 , SR a42 NHOR a42 , C(O)R b42 , C(O)NR c42 R d42 , C(O)NR c42 (OR a42)、C(O)OR a42 、OC(O)R b42 、OC(O)NR c42 R d42 、NR c42 R d42 、NR c42 NR c42 R d42 、NR c42 C(O)R b42 、NR c42 C(O)OR a42 、NR c42 C(O)NR c42 R d42 、C(=NR e42 )R b42 、C(=NR e42 )NR c42 R d42 、NR c42 C(=NR e42 )NR c42 R d42 、NR c42 C(=NR e42 )R b42 、NR c42 S(O)NR c42 R d42 、NR c42 S(O)R b42 、NR c42 S(O)2R b42 、NR c42 S(O)(=NR e42 )R b42 、NR c42 S(O)2NR c42 R d42 、S(O)R b42 、S(O)NR c42 R d42 、S(O)2R b42 、S(O)2NR c42 R d42 、OS(O)(=NR e42 )R b42 、OS(O)2R b42 、S(O)(=NR e42 )R b42 、SF5、P(O)R f42 R g42 、OP(O)(OR h42 )(OR i42 )、P(O)(OR h42 )(ORi42 ), and BR j42 R k42 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R G Substituents may be used, Each R 5A However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a51 , SR a51 NHOR a51 , C(O)R b51 , C(O)NR c51 R d51 , C(O)NR c51 (OR a51 ), C(O)OR a51 ,OC(O)R b51 , OC(O)NR c51 R d51 , NR c51 R d51 , NR c51 NR c51 R d51 , NR c51 C(O)Rb51 , NR c51 C(O)OR a51 , NR c51 C(O)NR c51 R d51 , C(=NR e51 )R b51 , C(=NR e51 )NR c51 R d51 , NR c51 C(=NR e51 )NR c51 R d51 , NR c51 C(=NR e51 )R b51 , NR c51 S(O)NR c51 R d51 , NR c51 S(O)R b51 , NR c51 S(O)2R b51 , NR c51 S(O)(=NR e51 )R b51 , NR c51 S(O)2NR c51 R d51 S(O)R b51 , S(O)NR c51 R d51 S(O)2R b51 , S(O)2NR c51 R d51 OS(O)(=NR e51 )R b51 OS(O)2R b51 , S(O)(=NR e51 )R b51 SF5, P(O)R f51 R g51 , OP(O)(OR h51 )(OR i51 ), P(O)(OR h51 )(OR i51 ), and BR j51 R k51 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5B Substituents may be used, Each R 5B However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a52 , SR a52 NHOR a52 , C(O)R b52 , C(O)NR c52 R d52 , C(O)NR c52 (OR a52 ), C(O)OR a52 ,OC(O)R b52 , OC(O)NR c52 R d52 , NR c52 R d52 , NR c52 NR c52 R d52 , NR c52 C(O)R b52 , NR c52 C(O)OR a52 , NR c52 C(O)NR c52 R d52 , C(=NR e52 )R b52 , C(=NR e52 )NRc52 R d52 , NR c52 C(=NR e52 )NR c52 R d52 , NR c52 C(=NR e52 )R b52 , NR c52 S(O)NR c52 R d52 , NR c52 S(O)R b52 , NR c52 S(O)2R b52 , NR c52 S(O)(=NR e52 )R b52 , NR c52 S(O)2NR c52 R d52 S(O)R b52 , S(O)NR c52 R d52 S(O)2R b52 , S(O)2NR c52 R d52 OS(O)(=NR e52 )R b52 OS(O)2R b52 , S(O)(=NR e52 )R b52 SF5, P(O)R f52 R g52 , OP(O)(OR h52 )(OR i52 ), P(O)(OR h52 )(OR i52 ), and BR j52 R k52 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4Each alkyl group can be one, two, three, or four independently selected R G Substituents may be used, Each R a4 , R c4 , and R d4 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 4A Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c4 and R d4 However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 10-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 10-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. 4A Substituents may be used, Each R b4 However, C 1~6Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 4A Substituents may be used, Each R e4 However, H, OH, CN, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f4 and R g4 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h4 and R i4 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R j4 and R k4 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j4 and R k4 However, along with the B atom to which they are bonded, C 1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R a41 , R c41 , and R d41 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 4B Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c41 and R d41 However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 7-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 7-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. 4B Substituents may be used, Each R b41 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 4B Substituents may be used, Each R e41 However, H, OH, CN, C 1~6 Alkyl, C1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f41 and R g41 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h41 and R i41 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each Rj41 and R k41 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j41 and R k41 However, along with the B atom to which they are bonded, C 1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R a42 , R c42 , and R d42 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R G Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c42 and R d42However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 7-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 7-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. G Substituents may be used, Each R b42 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. G Substituents may be used, Each R e42 However, H, OH, CN, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f42 and R g42 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h42 and R i42 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R j42 and R k42 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j42 and R k42 However, along with the B atom to which they are bonded, C 1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R a5 , R c5 , and R d5 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5A Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c5 and R d5 However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 10-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 10-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. 5A Substituents may be used, Each R b5 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5A Substituents may be used, Each R e5 However, H, OH, CN, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f5 and R g5 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h5 and R i5 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected independently of alkyl, Each R j5 and R k5 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j5 and R k5 However, along with the B atom to which they are bonded, C 1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R a51 , R c51 , and R d51 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5B Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c51 and R d51 However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 7-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 7-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. 5B Substituents may be used, Each R b51 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5B Substituents may be used, Each R e51 However, H, OH, CN, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f51 and R g51 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h51 and R i51 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R j51 and R k51 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j51 and R k51 However, along with the B atom to which they are bonded, C1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R a52 , R c52 , and R d52 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R G Substituents may be substituted, Alternatively, any of the R atoms bonded to the same N atom c52 and R d52 However, together with the N atom to which they are bonded, they form a 5 or 6-membered heteroaryl or 4- to 7-membered heterocycloalkyl group, and each of the 5 or 6-membered heteroaryl and 4- to 7-membered heterocycloalkyl groups can have one, two, three, or four independently selected R atoms. G Substituents may be used, Each R b52 However, C1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. G Substituents may be used, Each R e52 However, H, OH, CN, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R f52 and R g52 However, H, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Haloalkyl, C 1~6 Haloalkoxy, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R h52 and R i52 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, Each R j52 and R k52 However, OH, C 1~6 Alkoxy, and C 1~6 Selected independently from haloalkoxys, or any of the R atoms bonded to the same B atom j52 and R k52 However, along with the B atom to which they are bonded, C 1~6 Alkyl and C 1~6 Forming a 5- or 6-membered heterocycloalkyl group which is optionally substituted with one, two, three, or four substituents independently selected from the haloalkyl group, Each R G However, H, D, OH, NO2, CN, Halo, C 1~3 Alkyl, C 2~3 Alkenil, C 2~3 Alkinyl, C 1~3 Haloalkyl, cyano-C 1~3 Alkyl, HO-C 1~3 Alkyl, C 1~3 Alkoxy-C 1~3 Alkyl, C 3~7 Cycloalkyl, C 1~3 Alkoxy, C 1~3 Haloalkoxy, amino, C 1~3 Alkylamino, di(C 1~3Alkyl)amino, thio, C 1~3 Alkylthio, C 1~3 Alkyl sulfinyl, C 1~3 Alkyl sulfonyl, carbamyl, C 1~3 Alkylcarbamyl, di(C 1~3 Alkyl)carbamyl, carboxy, C 1~3 Alkylcarbonyl, C 1~3 Alkoxycarbonyl, C 1~3 Alkylcarbonyloxy, C 1~3 Alkylcarbonylamino, C 1~3 Alkoxycarbonylamino, C 1~3 Alkylaminocarbonyloxy, C 1~3 Alkylsulfonylamino, aminosulfonyl, C 1~3 Alkylaminosulfonyl, di(C 1~3 Alkyl)aminosulfonyl, aminosulfonylamino, C 1~3 Alkylaminosulfonylamino, di(C 1~3 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1~3 Alkylaminocarbonylamino, and di(C 1~3 It is independently selected from alkyl)aminocarbonylamino.

[0118] In some embodiments, n is an integer selected from 0, 1, 2, 3, or 4. Ring portion A is a monocyclic 3- to 7-membered cycloalkyl or a monocyclic 4- to 7-membered heterocycloalkyl. R 1 C 1~6 Haloalkyl, C 3~7 Selected from cycloalkyl and phenyl, each of which is one or two independently selected R 4 Substitutable by substituents, R 2 C 2~6 Alkyl and C 1~6 Selected from haloalkyl groups, R 3 C 1~6 Alkyl and C 1~6Selected from haloalkyls, or R 2 and R 3 However, together with the carbon atoms to which they are bonded, they form ring B. Ring B is a 3- to 7-membered cycloalkyl ring. Each R 4 , Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, OR a4 , and NR c4 R d4 Selected independently from, Each R a4 , R c4 , and R d4 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R 5 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C(O)R b5 , C(O)NR c5 R d5 , C(O)OR a5 S(O)2R b5 , and S(O)2NR c5 R d5 Selected independently from, Each R 5A , Haro, CN, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R a5 , R c5 , and R d5 H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4Selected independently of alkyl, the C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5A Substituents may be used, Each R b5 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5A Substitutions may be used.

[0119] In some embodiments, n is an integer selected from 0, 1, 2, 3, or 4. Ring portion A is a monocyclic 4-7 membered heterocycloalkyl, R 1 C 1~6 Haloalkyl, C 3~7 Selected from cycloalkyl and phenyl, each of which is one or two independently selected R 4 Substitutable by substituents, R 2 These are ethyl, propyl, isopropyl, and C 1~3 Selected from fluoroalkyl groups, R 3These are methyl, ethyl, propyl, isopropyl, and C 1~3 Selected from fluoroalkyl groups, or R 2 and R 3 However, together with the carbon atoms to which they are bonded, they form ring B. Ring B is a 3-4 membered cycloalkyl ring. Each R 4 C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R 5 C 1~6 Alkyl, C 1~6 Haloalkyl and S(O)2R b5 Selected independently from, Each R b5 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 A selection made independently from cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is one or two independently selected R 5A Substitutable by substituents, Each R 5A , Haro, CN, C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0120] In some embodiments, the compound is of formula (B-Ia) [ka] A compound of or a pharmaceutically acceptable salt thereof, where k is n-1.

[0121] In some embodiments, R 1 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is independently selected in groups of 1, 2, 3, 4, 5, or 6 R 4 It may be substituted by substituents.

[0122] In some embodiments, R 1 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, C 3~7 Cycloalkyl-C 1~3 Selected from alkyl, phenyl, 4-10 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is one or two independently selected R 4 It may be substituted by substituents.

[0123] Several embodiments, each R 4 , Haro, CN, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, OR a4 , C(O)R b4 , C(O)NR c4 R d4 , C(O)OR a4 ,OC(O)R b4 , OC(O)NR c4 R d4 , NR c4 R d4 , NR c4 C(O)R b4 , NR c4 C(O)OR a4 , NR c4 C(O)NR c4 R d4 , NR c4 S(O)2R b4 , NR c4 S(O)2NR c4 R d4 S(O)2R b4, and S(O)2NR c4 R d4 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl and C 1~6 Each haloalkyl can be one, two, three, or four independently selected R 4A Substitutions may be used.

[0124] In some embodiments, Each R 4A Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, OR a41 , SR a41 , C(O)R b41 , C(O)NR c41 R d41 , C(O)OR a41 ,OC(O)R b41 , OC(O)NR c41 R d41 , NR c41 R d41 , NR c41 C(O)R b41 , NR c41 C(O)OR a41 , NR c41 C(O)NR c41 R d41 , NR c41 S(O)2R b41 , NR c41 S(O)2NR c41 R d41 S(O)2R b41 , and S(O)2NR c41 R d41 Selected independently from, Each R a4 , R c4 , and R d4 H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 4A Substituents may be used, Each R b4 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 4A Substituents may be used, Each R a41 , R c41 , and R d41 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R b41 C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0125] In some embodiments, Each R 4A, Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, OR a41 , C(O)R b41 , C(O)NR c41 R d41 , C(O)OR a41 , NR c41 R d41 , NR c41 C(O)R b41 NNR c41 S(O)2R b41 S(O)2R b41 , and S(O)2NR c41 R d41 Selected independently from, Each R a4 , R c4 , and R d4 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, the C 1~6 Alkyl and C 1~6 Each haloalkyl can be one, two, three, or four independently selected R 4A Substituents may be used, Each R b4 C 1~6 Alkyl and C 1~6 A haloalkyl group is independently selected, and each of them is independently selected by one, two, three, or four R groups. 4A Substituents may be used, Each R a41 , R c41 , and R d41 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R b41 C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0126] In some embodiments, the ring portion A is a monocyclic 3- to 7-membered cycloalkyl or a monocyclic 4- to 7-membered heterocycloalkyl.

[0127] In some embodiments, the ring portion A is a monocyclic 4- to 7-membered heterocycloalkyl group.

[0128] In some embodiments, the ring portion A is an azetidine ring, a pyrrolidine ring, a piperidine ring, or an azepane ring.

[0129] In some embodiments, the ring portion A is a piperidine ring.

[0130] In some embodiments, n is 1 or 2.

[0131] Several embodiments, each R 5 Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, OR a5 , SR a5 , C(O)R b5 , C(O)NR c5 R d5 , C(O)OR a5 ,OC(O)R b5 , OC(O)NR c5 R d5 , NR c5 R d5 , NR c5 C(O)R b5 , NR c5 C(O)OR a5 , NR c5 C(O)NR c5 R d5 , NR c5 S(O)2R b5 , NR c5 S(O)2NR c5 R d5 S(O)2R b5 , and S(O)2NR c5 R d5 It is selected independently of others.

[0132] In some embodiments, Each R a5 , R c5 , and Rd5 H and C 1~6 Selected independently of alkyl, Each R b5 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 A alkyl group is independently selected, and each of them is one or two independently selected R groups. 5A Substitutions may be used.

[0133] Several embodiments, each R 5 This is Halo and C 1~6 It is selected independently of alkyl.

[0134] Several embodiments, each R b5 C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5A Substitutions may be used.

[0135] In some embodiments, R b5 C 1~6 Alkyl, C3~6 Selected from cycloalkyl, phenyl, 4-6 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is a halo, C 1~6 One or two R groups independently selected from alkyl and 4-6 member heterocycloalkyl groups. 5A The 4-6 member heterocycloalkyl group is optionally substituted by a substituent, and the C 1~3 One or two R components selected independently of alkyl groups 5B It may be substituted by substituents.

[0136] In some embodiments, Each R 5 Hello, C 1~3 Alkyl, C 1~3 Haloalkyl, OR a5 , and NR c5 R d5 Selected independently from, Each R a5 , R c5 , and R d5 H and C 1~6 Selected independently of alkyl, R b5 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is one or two independently selected R 5A Substituents may be used, Each R 5A , Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, ORa51 , SR a51 , C(O)R b51 , C(O)NR c51 R d51 , C(O)OR a51 ,OC(O)R b51 , OC(O)NR c51 R d51 , NR c51 R d51 , NR c51 C(O)R b51 , NR c51 C(O)OR a51 , NR c51 C(O)NR c51 R d51 , NR c51 S(O)2R b51 , NR c51 S(O)2NR c51 R d51 S(O)2R b51 , and S(O)2NR c51 R d51 Selected independently from, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl are each one or two independently selected R 5B Substituents may be used, Each R a51 , R c51 , and R d51 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, the C 1~6 Alkyl and C 1~6 Each haloalkyl is one or two independently selected R 5B Substituents may be used, Each R b51 C 1~6 Alkyl and C 1~6 A haloalkyl group is independently selected, and each of them is one or two independently selected R groups. 5B Substituents may be used, Each R 5B , Haro, CN, C 1~6 Alkyl and C1~6 It is independently selected from haloalkyl groups.

[0137] In some embodiments, Each R 5 This is Halo and C 1~3 Selected independently of alkyl, R b5 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Selected from cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is one or two independently selected R 5A Substituents may be used, Each R 5A Hello, C 1~6 A C is independently selected from alkyl and 4- to 7-membered heterocycloalkyl groups. 1~6 Alkyl and 4- to 7-membered heterocycloalkyl groups are each selected from one or two independently. 5B Substituents may be used, Each R 5B C 1~6 It is selected independently of alkyl.

[0138] In some embodiments, the compound is of formula (B-II): [ka] A compound of or a pharmaceutically acceptable salt thereof, where the variables are defined according to the definitions provided herein.

[0139] In some embodiments, the compound is of formula (B-IIa): [ka] A compound of or a pharmaceutically acceptable salt thereof, where k is n-1 and the remaining variables are defined according to the definitions provided herein.

[0140] In some embodiments, the compound is of formula (B-IIb): [ka] A compound of or a pharmaceutically acceptable salt thereof, where k is n-1 and the remaining variables are defined according to the definitions provided herein.

[0141] In some embodiments, ring B is a 3- to 7-membered cycloalkyl ring.

[0142] In some embodiments, the compound is of formula (B-IIc): [ka] A compound of or a pharmaceutically acceptable salt thereof, where k is n-1 and the remaining variables are defined according to the definitions provided herein.

[0143] In some embodiments, the compound is of formula (B-IId): [ka] A compound of or a pharmaceutically acceptable salt thereof, in which, X is a bond or CH2, Y is a bond or CH2, k is n-1.

[0144] In some embodiments, the compound has the formula (B-Ia), where, k is n-1, n is an integer selected from 1 and 2. Ring portion A is a monocyclic 4-6 member heterocycloalkyl, R 1 However, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, phenyl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected from alkyl groups, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, phenyl, 4-10 member heterocycloalkyl, 5-10 member heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Each alkyl group consists of one, two, or three independently selected R 4 Substitutable by substituents, R 2 and R 3 However, together with the carbon atoms to which they are bonded, they form ring B. Ring B is a 3- to 7-membered cycloalkyl ring, Each R 4 However, H, Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~4 Cycloalkyl, OR a4 , C(O)R b4 , C(O)NR c4 R d4 , C(O)OR a4 ,OC(O)R b4 , OC(O)NR c4 R d4 , NR c4 R d4 , NR c4 C(O)R b4 , NR c4 C(O)OR a4 , NR c4 C(O)NR c4 R d4 , NR c4 S(O)2R b4 , NR c4 S(O)2NRc4 R d4 S(O)2R b4 , and S(O)2NR c4 R d4 Selected independently from, Each R 5 However, H, Haro, CN, C 1~3 Alkyl and C 1~3 Selected independently from haloalkyl groups, Each R 5A However, H, D, Haro, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl, 5-6 member heteroaryl-C 1~4 Alkyl, OR a51 , C(O)R b51 , C(O)NR c51 R d51 C(O)OR a51 ,OC(O)R b51 , OC(O)NR c51 R d51 , NR c51 R d51 , NR c51 C(O)R b51 , NR c51 C(O)OR a51 , NR c51 C(O)NR c51 R d51 , NR c51 S(O)2R b51 , NR c51 S(O)2NR c51 R d51 S(O)2R b51 , and S(O)2NR c51 R d51 Selected independently from, C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5B Substituents may be used, Each R 5B However, H, Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, OH, NO2, CN, Halo, C 1~3 Alkyl, C 2~3 Alkenil, C 2~3 Alkinyl, C 1~3 Haloalkyl, cyano-C 1~3 Alkyl, HO-C 1~3 Alkyl, C 1~3 Alkoxy-C 1~3 Alkyl, C 3~7 Cycloalkyl, C 1~3 Alkoxy, C 1~3 Haloalkoxy, amino, C 1~3 Alkylamino, di(C 1~3 Alkyl)amino, thio, C 1~3 Alkylthio, C 1~3 Alkyl sulfinyl, C 1~3 Alkyl sulfonyl, carbamyl, C 1~3 Alkylcarbamyl, di(C 1~3 Alkyl)carbamyl, carboxy, C 1~3 Alkylcarbonyl, C 1~3 Alkoxycarbonyl, C 1~3 Alkylcarbonyloxy, C 1~3 Alkylcarbonylamino, C 1~3 Alkoxycarbonylamino, C 1~3 Alkylaminocarbonyloxy, C 1~3 Alkylsulfonylamino, aminosulfonyl, C 1~3 Alkylaminosulfonyl, di(C1~3 Alkyl)aminosulfonyl, aminosulfonylamino, C 1~3 Alkylaminosulfonylamino, di(C 1~3 Alkyl)aminosulfonylamino, aminocarbonylamino, C 1~3 Alkylaminocarbonylamino, and di(C 1~3 Independently selected from alkyl)aminocarbonylamino, Each R a4 , R c4 , and R d4 However, H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, Each R b5 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C 3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5A Substituents may be used, Each R a51 , R c51 , and R d51 However, H, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C1~4 Selected independently of alkyl, the C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Each alkyl group can be one, two, three, or four independently selected R 5B Substituents may be used, Each R b51 However, C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C 1~4 Alkyl and 5-6 member heteroaryl-C 1~4 R is independently selected from alkyl groups, each of which is one, two, three, or four independently selected R groups. 5B Substitutions may be used.

[0145] In some embodiments of the compound of formula (B-Ia), k is n-1, n is either 1 or 2. Ring portion A is a 4-6 member heterocycloalkyl group. R 1 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~10 Cycloalkyl, 6-10 membered aryl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, C3~10 Cycloalkyl-C 1~4 Alkyl, 6-10 member aryl-C 1~4 Alkyl, 4-10 member heterocycloalkyl-C 1~4 Alkyl and 5-10 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is one, two, or three independently selected R groups. 4 Substitutable by substituents, Each R 4 , Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, OR a4 , and NR c4 R d4 Selected independently from, Each R a4 , R c4 , and R d4 H and C 1~6 Selected independently of alkyl, R 2 and R 3 They form ring B together with the carbon atoms to which they are bonded. Ring B is a 3-4 membered cycloalkyl ring. Each R 5 Hello, C 1~3 Alkyl, C 1~3 Haloalkyl, OR a5 , and NR c5 R d5 Selected independently from, Each R a5 , R c5 , and R d5 H and C 1~6 Selected independently of alkyl, R b5 C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, C 3~7 Cycloalkyl-C 1~4 Alkyl, phenyl-C 1~4 Alkyl, 4-7 member heterocycloalkyl-C1~4 Alkyl and 5-6 member heteroaryl-C 1~4 Selected from alkyl groups, each of which is one or two independently selected R 5A Substituents may be used, Each R 5A , Haro, CN, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, 5-6 member heteroaryl, OR a51 , SR a51 , C(O)R b51 , C(O)NR c51 R d51 , C(O)OR a51 ,OC(O)R b51 , OC(O)NR c51 R d51 , NR c51 R d51 , NR c51 C(O)R b51 , NR c51 C(O)OR a51 , NR c51 C(O)NR c51 R d51 , NR c51 S(O)2R b51 , NR c51 S(O)2NR c51 R d51 S(O)2R b51 , and S(O)2NR c51 R d51 Selected independently from, C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, phenyl, 4-7 member heterocycloalkyl, and 5-6 member heteroaryl are each one or two independently selected R 5B Substituents may be used, Each R a51 , R c51 , and R d51 H, C 1~6 Alkyl and C 1~6 Selected independently from haloalkyl groups, the C 1~6 Alkyl and C 1~6Each haloalkyl is one or two independently selected R 5B Substituents may be used, Each R b51 C 1~6 Alkyl and C 1~6 A haloalkyl group is independently selected, and each of them is one or two independently selected R groups. 5B Substituents may be used, Each R 5B , Haro, CN, C 1~6 Alkyl and C 1~6 It is independently selected from haloalkyl groups.

[0146] In some embodiments of the compound of formula (B-Ia), k is n-1, n is either 1 or 2. Ring portion A is a piperidine ring, R 1 C 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 Cycloalkyl, C 3~7 Cycloalkyl, C 3~7 Cycloalkyl-C 1~3 Selected from alkyl, phenyl, 4-10 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is one or two independently selected R 4 Substitutable by substituents, Each R 4 Halo, OH, C 1~3 Alkyl and C 1~3 Selected independently of alkoxy, R 2 and R 3 They form ring B together with the carbon atoms to which they are bonded. Ring B is a 3-4 membered cycloalkyl ring. Each R 5 This is Halo and C 1~3 Selected independently of alkyl, R b5 C 1~6 Alkyl, C 3~6Selected from cycloalkyl, phenyl, 4-6 member heterocycloalkyl, and 5-6 member heteroaryl, each of which is a halo, C 1~6 One or two R groups independently selected from alkyl and 4-6 member heterocycloalkyl groups. 5A The 4-6 member heterocycloalkyl group is optionally substituted by a substituent, and the C 1~3 One or two R components selected independently of alkyl groups 5B It may be substituted by substituents.

[0147] In some embodiments, the compound is a compound selected from the compounds of the examples, or a pharmaceutically acceptable salt thereof.

[0148] In some embodiments, the substituents described herein include "alkyl," "alkenyl," "alkynyl," "aryl," "phenyl," "cycloalkyl," "heterocycloalkyl," or "heteroaryl" substituents or "-C" substituents. 1~4 The one, two, three, four, five, six, seven, or eight hydrogen atoms bonded to the carbon atoms of the alkyl- and alkylene linking groups may be replaced by deuterium atoms.

[0149] For clarity, it is further understood that certain features of the present invention, described in relation to separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the present invention, described in relation to a single embodiment for brevity, may also be provided separately or in any preferred partial combination.

[0150] Divalent linked substituents are described in various places in this specification. Each divalent linked substituent is specifically intended to include both the forward and backward forms of the linked substituent. For example, -NR(CR'R'') n - is -NR(CR'R'') n -and-(CR'R'') nThis includes both NR- and NR-. If the structure explicitly requires a linking group, the Markush variables listed for that group are understood to be linking groups.

[0151] "One R" 5 S(O)2R b5 And each of the remaining R 5 Embodiments that state "the is independently selected from the following" are those that do not use -S(O)2R b5 When a substituent is shown in a formula and combined with multiple dependents, the -S(O)2R that is not formally bonded is called -S(O)2R. b5 Due to substituents, "one R 5 S(O)2R b5 The phrase "and" is replaced. In such embodiments, when combined with an expression having an integer k, R 5 (n possible R of substituents) 5 One of the substituents is S(O)2R in the formula b5 Replaced by the remaining R 5 Substituents (k remaining R 5 Each of the substituents is "the remaining R 5 It is selected independently from the list of "".

[0152] The term "n-membered," where n is an integer, typically describes the number of ring-forming atoms in a part of the ring where the number of ring-forming atoms is n. For example, piperidinyl is an example of a 6-membered heterocycloalkyl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, pyridyl is an example of a 6-membered heteroaryl ring, and 1,2,3,4-tetrahydronaphthalene is an example of a 10-membered cycloalkyl group.

[0153] As used herein, the phrase "optionally substituted" means either unsubstituted or substituted. Substituents are selected independently, and substitution may occur at any chemically accessible position. As used herein, the term "substituted" means that a hydrogen atom is removed and replaced by a substituent. A single divalent substituent, e.g., an oxo, can replace two hydrogen atoms. It should be understood that substitutions at a given atom are limited by valence, that the normal valence of the specified atom is not exceeded, and that the substitution results in a stable compound.

[0154] As used herein, the phrase “each ‘variable’ is independently selected from the following” means substantially the same as “each instance of ‘variable’ is selected from the following.”

[0155] Any variable (for example, R S If a compound has two or more R groups, the definition of that group for each occurrence is independent of the definition of that group for all other occurrences. For example, if a compound has one, two, three, or four R groups... S If it is shown to be substituted by, then the base has up to four R S The base may be substituted depending on the case, and each occurrence of R S R S The selection is independent of the definition. Furthermore, combinations of substituents and / or variables are permitted only if such combinations result in stable compounds. For example, the combination of a first M group and a second M group in a combination of two R groups is permitted only if such an MM combination results in a stable compound (for example, MM is not permitted if it would form a highly reactive compound such as a peroxide with an OO bond).

[0156] Through the definition, "C n~m The term "C" indicates a range including the endpoints, where n and m are integers representing the number of carbon atoms. For example, C 1~3 , C 1~4 , C1~6 These are some examples.

[0157] When used herein, "C" is used alone or in combination with other terms. n~m The term "alkyl" refers to a saturated hydrocarbon group having n to m carbon atoms, which may be linear or branched. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl (Me), ethyl (Et), n-propyl (n-Pr), isopropyl (iPr), n-butyl, tert-butyl, isobutyl, and sec-butyl, and higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, and 1,2,2-trimethylpropyl. In some embodiments, the alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

[0158] When used herein, "C n~m An "alkenyl" refers to an alkyl group having one or more double carbon-carbon bonds and n to m carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, and sec-butenyl. In some embodiments, the alkenyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms.

[0159] When used herein, "C n~m "Alkynyl" refers to an alkyl group having one or more triple carbon-carbon bonds and n to m carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl, propyne-1-yl, and propyne-2-yl. In some embodiments, the alkynyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms. As used herein, "C" is used alone or in combination with other terms. n~mThe term "alkoxy" refers to a group of the formula -O-alkyl, where the alkyl group has n to m carbon atoms. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and butoxy (e.g., n-butoxy and tert-butoxy). In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0160] As used herein, the term "amino" refers to the group of formula -NH2.

[0161] As used herein, the term “aryl,” when used alone or in combination with other terms, refers to an aromatic hydrocarbon group, which may be monocyclic or polycyclic (e.g., having two fused rings). n~m The term "aryl" refers to an aryl group having n to m ring carbon atoms. Examples of aryl groups include phenyl, naphthyl, anthracenyl, phenantrenyl, indanyl, and indenyl. In some embodiments, the aryl group has 6 to 10 carbon atoms. In some embodiments, the aryl group is phenyl or naphthyl. In some embodiments, the aryl is phenyl.

[0162] As used herein, “halo” refers to F, Cl, Br, or I. In some embodiments, halo is F, Cl, or Br. In some embodiments, halo is F or Cl. In some embodiments, halo is F. In some embodiments, halo is Cl.

[0163] When used herein, "C n~m A "haloalkoxy" refers to a -O-haloalkyl group having n to m carbon atoms. Exemplary haloalkoxy groups include OCF3 and OCHF2. In some embodiments, the haloalkoxy group is fluorinated only. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0164] When used herein, "C" is used alone or in combination with other terms. n~m The term "haloalkyl" refers to an alkyl group having one halogen atom ~2s+1 halogen atoms (which may be the same or different), where "s" is the number of carbon atoms in the alkyl group, and the alkyl group has n ~ m carbon atoms. In some embodiments, the haloalkyl group is fluorinated only. In some embodiments, the alkyl group has 1 ~ 6, 1 ~ 4, or 1 ~ 3 carbon atoms. Exemplary haloalkyl groups include CF3, C2F5, CHF2, CH2F, CCl3, CHCl2, and C2Cl5.

[0165] As used herein, the term "thio" refers to the group of formula -SH.

[0166] When used herein, "C n~m The term "alkylamino" refers to a group of the formula -NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0167] When used herein, "C n~m The term "alkoxycarbonyl" refers to a group of the formula -C(O)O-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0168] When used herein, "C n~m The term "alkylcarbonyl" refers to a group of the formula -C(O)-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0169] When used herein, "C n~mThe term "alkylcarbonylamino" refers to a group of the formula -NHC(O)-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0170] When used herein, "C n~m The term "alkoxycarbonylamino" is derived from the formula -NHC(O)O(C n~m This refers to an alkyl group, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0171] When used herein, "C n~m The term "alkylsulfonylamino" refers to a group of the formula -NHS(O)2-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0172] As used herein, the term "aminosulfonyl" refers to the group of the formula -S(O)2NH2.

[0173] When used herein, "C n~m The term "alkylaminosulfonyl" refers to a group of the formula -S(O)2NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0174] When used herein, "Ji (C) n~m The term "alkyl)aminosulfonyl" refers to a group of the formula -S(O)2N(alkyl)2, where each alkyl group independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0175] As used herein, the term "aminosulfonylamino" refers to the group of the formula -NHS(O)2NH2.

[0176] When used herein, "C n~m The term "alkylaminosulfonylamino" refers to a group of the formula -NHS(O)2NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0177] When used herein, "Ji (C) n~m The term "alkyl)aminosulfonylamino" refers to a group of the formula -NHS(O)2N(alkyl)2, where each alkyl group independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0178] As used herein, the term "aminocarbonylamino," when used alone or in combination with other terms, refers to the group of the formula -NHC(O)NH2.

[0179] When used herein, "C n~m The term "alkylaminocarbonylamino" refers to a group of the formula -NHC(O)NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0180] When used herein, "Ji (C) n~m The term "alkyl)aminocarbonylamino" refers to a group of the formula -NHC(O)N(alkyl)2, where each alkyl group independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0181] When used herein, "C n~mThe term "alkylcarbamyl" refers to a group of the formula -C(O)-NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0182] When used herein, "C n~m The term "alkylthio" refers to a group of the formula -S-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0183] When used herein, "C n~m The term "alkylsulfinyl" refers to a group of the formula -S(O)-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0184] When used herein, "C n~m The term "alkylsulfonyl" refers to a group of the formula -S(O)2-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0185] When used herein, "Cyan-C" 1~6 The term "alkyl" is derived from the formula -(C 1~6 Refers to the alkylene-CN group. As used herein, "cyano-C 1~3 The term "alkyl" is derived from the formula -(C 1~3 This refers to the alkylene-CN group.

[0186] When used herein, "HO-C 1~6 The term "alkyl" is derived from the formula -(C 1~6 Refers to the alkylene-OH group. When used herein, "HO-C 1~3 The term "alkyl" is derived from the formula -(C 1~3 This refers to the alkylene-OH group.

[0187] When used herein, "C 1~6 Alkoxy-C 1~6 The term "alkyl" is derived from the formula -(C 1~6 Alkylene)-O(C) 1~6 Refers to the alkyl group. When used herein, "C 1~3 Alkoxy-C 1~3 The term "alkyl" is derived from the formula -(C 1~3 Alkylene)-O(C) 1~3 It refers to the alkyl group.

[0188] As used herein, the term "carboxyl" refers to the group of the formula -C(O)OH.

[0189] When used herein, "Ji (C) n~m The term "-alkyl)amino" refers to a group of the formula -N(alkyl)2, where each of the two alkyl groups independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0190] When used herein, "Ji (C) n~m The term "-alkyl)carbamyl" refers to a group of the formula -C(O)N(alkyl)2, where each of the two alkyl groups independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0191] When used herein, "C n~m The term "alkylcarbonyloxy" refers to a group of the formula -OC(O)-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0192] As used herein, "aminocarbonyloxy" refers to the group of the formula -OC(O)-NH2.

[0193] When used herein, "C n~m "Alkylaminocarbonyloxy" is a group of the formula -OC(O)-NH-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0194] When used herein, "Ji (C) n~m "Alkyl)aminocarbonyloxy" is a group of the formula -OC(O)-N(alkyl)2, where each alkyl group independently has n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0195] When used herein, C n~m Alkoxycarbonylamino refers to a group of the formula -NHC(O)-O-alkyl, where the alkyl group has n to m carbon atoms.

[0196] As used herein, the term "carbamyl" refers to the group of the formula -C(O)NH2.

[0197] As used herein, the term "carbonyl," when used alone or in combination with other terms, refers to a -C(O)- group.

[0198] As used herein, “cycloalkyl” refers to non-aromatic cyclic hydrocarbons containing cyclized alkyl and alkenyl groups. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having two, three, or four fused rings) groups, spiro rings, and crosslinking rings (e.g., crosslinked bicycloalkyl groups). The ring-forming carbon atoms of cycloalkyl groups may optionally be substituted with oxo or sulfide (e.g., C(O) or C(S)). The definition of cycloalkyl also includes moieties having one or more aromatic rings fused to (i.e., sharing a common bond with) a cycloalkyl ring, such as benzo or thienyl derivatives like cyclopentane and cyclohexane. Cycloalkyl groups containing fused aromatic rings may be bonded by any ring-forming atoms, including the ring-forming atoms of the fused aromatic ring. Cycloalkyl groups may have 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring-forming carbons (i.e., C 3~14 ) may have. In some embodiments, the cycloalkyl is optionally substituted with CH2F, CHF2, CF3, and CF2CF3. 3~12 It is a monocyclic or bicyclic cycloalkyl. In some embodiments, the cycloalkyl is C 3~10 It is a monocyclic or bicyclic cycloalkyl. In some embodiments, the cycloalkyl is C 3~7 It is a monocyclic cycloalkyl. In some embodiments, the cycloalkyl is C 4~7 It is a monocyclic cycloalkyl. In some embodiments, the cycloalkyl is C 4~14The cycloalkyl group is a spiro ring or a crosslinked cycloalkyl group (e.g., a crosslinked bicycloalkyl group). Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, cubane, adamantane, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, bicyclo[2.2.2]octanyl, and spiro[3.3]heptanyl. In some embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0199] As used herein, “heteroaryl” refers to a monocyclic or polycyclic (e.g., having two, three, or four fused rings) aromatic heterocycle having at least one heteroatom ring member selected from N, O, S, and B. In some embodiments, the heteroaryl ring has one, two, three, or four heteroatom ring members independently selected from N, O, S, and B. In some embodiments, any of the ring-forming N in the heteroaryl moiety may be an N-oxide. In some embodiments, the heteroaryl is a 5- to 10-membered monocyclic or bicyclic heteroaryl having one, two, three, or four heteroatom ring members independently selected from N, O, and S. In some embodiments, the heteroaryl is a 5- to 10-membered monocyclic or bicyclic heteroaryl having one, two, three, or four heteroatom ring members independently selected from N, O, and S. In some embodiments, the heteroaryl is a 5- to 10-membered monocyclic or bicyclic heteroaryl having one, two, three, or four heteroatom ring members independently selected from N, O, and S. In some embodiments, the heteroaryl is a 5- to 6-membered monocyclic heteroaryl having one or two heteroatom ring members independently selected from N, O, S, and B. In some embodiments, the heteroaryl is a 5- to 6-membered monocyclic heteroaryl having one or two heteroatom ring members independently selected from N, O, and S. In some embodiments, the heteroaryl group contains 3- to 14, 3- to 10, 4- to 14, 4- to 10, 3- to 7, or 5- to 6 ring-forming atoms. In some embodiments, the heteroaryl group has 1- to 4 ring-forming heteroatoms, 1- to 3 ring-forming heteroatoms, 1- to 2 ring-forming heteroatoms, or 1 ring-forming heteroatom. If the heteroaryl group contains two or more heteroatom ring members, the heteroatoms may be the same or different.Examples of heteroaryl groups include, but are not limited to, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, pyrazole, azolyl, oxazole, isoxazole, thiazole, isothiazole, imidazole, furan, thiophene, triazole, tetrazole, thiadiazole, quinoline, isoquinoline, indole, benzothiophene, benzofuran, benzoisoxazole, imidazo[1,2-b]thiazole, purine, triazine, thieno[3,2-b]pyridine, imidazo[1,2-a]pyridine, 1,5-naphthyridine, and 1H-pyrazolo[4,3-b]pyridine.

[0200] A five-membered heteroaryl is a heteroaryl group having five ring-forming atoms, where one or more of the ring-forming atoms (e.g., one, two, or three) are independently selected from N, O, S, or B. Exemplary five-membered ring heteroaryls are thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, 1,3,4-oxadiazolyl, and 1,2-dihydro-1,2-azavolin.

[0201] A six-membered heteroaryl ring is a heteroaryl group having six ring-forming atoms, where one or more of the ring-forming atoms (e.g., one, two, or three) are independently selected from N, O, S, and B. Exemplary six-membered heteroaryl rings are pyridyl, pyrazinyl, pyrimidinyl, triazinyl, and pyridazinyl.

[0202] As used herein, “heterocycloalkyl” refers to a monocyclic or polycyclic heterocycle having at least one non-aromatic ring (saturated or partially unsaturated ring), wherein one or more of the ring-forming carbon atoms of the heterocycloalkyl are replaced by heteroatoms selected from N, O, S, and B, and the ring-forming carbon atoms and heteroatoms of the heterocycloalkyl may optionally be replaced by one or more oxo or sulfide atoms (e.g., C(O), S(O), C(S), or S(O)2). Examples of heterocycloalkyls include monocyclic and polycyclic (e.g., having two fused rings) systems. Heterocycloalkyls include monocyclic and polycyclic 12, 4-12, 3-10, 4-10, 3-7, 4-7, and 5-6 membered heterocycloalkyls. Other examples of heterocycloalkyl groups include spiro rings and crosslinking rings (e.g., 5-14 membered crosslinking biheterocycloalkyl rings having one or more ring-forming carbon atoms substituted by heteroatoms independently selected from N, O, S, and B). Heterocycloalkyl groups may be bonded by ring-forming carbon atoms or ring-forming heteroatoms. In some embodiments, the heterocycloalkyl group contains 0-3 double bonds. In some embodiments, the heterocycloalkyl group contains 0-2 double bonds.

[0203] Furthermore, the definition of heterocycloalkyl includes moieties having one or more aromatic rings condensed (i.e., sharing a common bond) with a non-aromatic heterocycle, such as benzo or thienyl derivatives like piperidine, morpholine, and azepine. Heterocycloalkyl groups containing condensed aromatic rings may be bonded by any ring-forming atoms, including the ring-forming atoms of the condensed aromatic ring. In some embodiments, heterocycloalkyl groups contain 3 to 14 ring-forming atoms, 4 to 14 ring-forming atoms, 3 to 10 ring-forming atoms, 4 to 10 ring-forming atoms, 3 to 7 ring-forming atoms, or 5 to 6 ring-forming atoms. In some embodiments, heterocycloalkyl groups have 1 to 4 heteroatoms, 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom. In some embodiments, heterocycloalkyl groups are monocyclic 4 to 6-membered heterocycloalkyl groups having one or two heteroatoms independently selected from N, O, S, and B, and having one or more oxidized ring members.

[0204] Examples of heterocycloalkyl groups include pyrrolidine-2-one, 1,3-isoxazolidine-2-one, pyranyl, tetrahydropyran, oxetanyl, azetidinyl, morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, and imi. Dazolidinil, azepanil, benzazapene, 1,2,3,4-tetrahydroisoquinoline, azabicyclo[3.1.0]hexanil, diazabicyclo[3.1.0]hexanil, oxabicyclo[2.1.1]hexanil, azabicyclo[2.2.1]heptanil, diazabicyclo[2.2.1]heptanil, azabicyclo[3.1.1]heptanil, diazabicyclo[3.1.1]heptanil, azabicyclo [3.2.1] Octanil, diazabicyclo[3.2.1] Octanil, oxabicyclo[2.2.2] Octanil, azaabicyclo[2.2.2] Octanil, azaadamantanil, diazaadamantanil, oxa-adamantanil, azaspiro[3.3] Heptanil, diazaspiro[3.3] Heptanil, oxa-azaspiro[3.3] Heptanil, azaspiro[3.4] Octanil, diazaspiro[3.4 Examples include octanyl, oxa-azaspiro[3.4]octanyl, azaspiro[2.5]octanyl, diazaspiro[2.5]octanyl, azaspiro[4.4]nonanyl, diazaspiro[4.4]nonanyl, oxa-azaspiro[4.4]nonanyl, azaspiro[4.5]decanyl, diazaspiro[4.5]decanyl, diazaspiro[4.4]nonanyl, and oxa-diazaspiro[4.4]nonanyl.

[0205] When used herein, "C o~p Cycloalkyl-C n~m "Alkyl-" refers to the group of the formula cycloalkyl-alkylene-, where the cycloalkyl group has 0 to p carbon atoms and the alkylene linking group has n to m carbon atoms.

[0206] When used herein, "C o~p Aryl-C n~m"Alkyl-" refers to the group of the formula aryl-alkylene-, where the aryl group has 0 to p carbon atoms and the alkylene linking group has n to m carbon atoms.

[0207] When used herein, "heteroaryl-C" n~m The "alkyl-" designation refers to the group of the formula heteroaryl-alkylene-, where the alkylene linking group has n to m carbon atoms.

[0208] When used herein, "heterocycloalkyl-C" n~m "Alkyl-" refers to the group of the formula heterocycloalkyl-alkylene-, where the alkylene linking group has n to m carbon atoms.

[0209] As used herein, the term "alkylene" refers to a divalent linear or branched alkyl linking group. Examples of "alkylene groups" include methylene, ethane-1,1-diyl, ethane-1,2-diyl, propane-1,3-diyl, propane-1,2-diyl, and propane-1,1-diyl.

[0210] As used herein, the term "alkenylene" refers to a divalent linear or branched alkenyl linking group. Examples of "alkenylene groups" include ethene-1,1-diyl, ethene-1,2-diyl, propene-1,3-diyl, 2-butene-1,4-diyl, 3-pentene-1,5-diyl, 3-hexene-1,6-diyl, and 3-hexene-1,5-diyl.

[0211] As used herein, the term "alkynylene" refers to a divalent linear or branched alkynyl linking group. Examples of "alkynylene groups" include propyne-1,3-diyl, 2-butyn-1,4-diyl, 3-pentyn-1,5-diyl, 3-hexyn-1,6-diyl, and 3-hexyn-1,5-diyl.

[0212] As used herein, the term "oxo" refers to an oxygen atom as a divalent substituent (i.e., =O) that, when bonded to carbon, forms a carbonyl group (e.g., C=O or C(O)), or when bonded to a nitrogen or sulfur heteroatom, forms a nitroso, sulfinyl, or sulfonyl group.

[0213] As used herein, the term “independently selected from” means that each occurrence of a variable or substituent is independently selected from the applicable list for each occurrence.

[0214] In certain places, definitions or embodiments refer to specific rings (e.g., azetidine rings, pyridine rings, etc.). Unless otherwise indicated, these rings may be bonded to any ring member, provided that the valence of the atoms does not exceed the limit. For example, an azetidine ring may be bonded at any position on the ring, whereas a pyridine-3-yl ring is bonded at position 3.

[0215] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods relating to preparing optically active forms from optically inert starting materials are known in the art and include methods such as the separation of racemic mixtures or stereoselective synthesis. Many geometric isomers, such as olefins and C=N double bonds, may also be present in the compounds described herein, and all such stable isomers are intended in the present invention. Cis and trans geometric isomers of the compounds of this disclosure are described, and these may be isolated as mixtures of isomers or as separate isomeric forms. In some embodiments, the compounds have an (R) configuration. In some embodiments, the compounds have an (S) configuration. Formulas provided herein (e.g., formulas (AI), (BI), etc.) include stereoisomers of the compounds.

[0216] The separation of racemic mixtures of compounds can be carried out by any of a number of methods known in the art. An exemplary method is fractional recrystallization using chiral dividing acids, which are optically active salt-forming organic acids. Suitable dividing agents for fractional recrystallization are, for example, optically active acids, such as D and L isomers of various optically active camphorsulfonic acids, including tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or β-camphorsulfonic acid. Other suitable dividing agents for fractional crystallization include stereoisomerically pure forms of α-methylbenzylamine (e.g., S and R isomers, or diastereomerically pure forms), and stereoisomerically pure forms of 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, and 1,2-diaminocyclohexane.

[0217] The separation of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). A suitable elution solvent composition can be determined by those skilled in the art.

[0218] The compounds provided herein also include tautomers. Tautomerism results from the exchange of a single bond with an adjacent double bond, along with the associated transfer of protons. Examples of tautomers include prototropic tautomers, which are isomeric protonated states having the same empirical formula and total charge. Exemplary examples of prototropic tautomers include ketone-enol pairs, amide-imido acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in the heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4-triazoles, 1H- and 2H-isoindole, 2-hydroxypyridine and 2-pyridone, and 1H- and 2H-pyrazoles. Tautomers may be in equilibrium or may be sterically fixed into one form by appropriate substitution. Compounds herein identified by name or structure as a specific tautomer, unless otherwise specified, are intended to include other tautomers.

[0219] All compounds, and their pharmaceutically acceptable salts, can be found together with other substances such as water and solvents (e.g., hydrates and solvates), or they can be isolated.

[0220] In some embodiments, the preparation of the compound may involve adding an acid or base that affects, for example, the catalytic action of the desired reaction or the formation of a salt form such as an acid addition salt.

[0221] In some embodiments, the compounds or salts thereof provided herein are substantially isolated. "Substantially isolated" means that the compounds are at least partially, or substantially, separated from the environment in which they were formed or detected. Partial isolation may include, for example, a concentrated composition of the compounds provided herein. Substantial isolation may include a composition containing at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of the compounds or salts thereof provided herein. Methods for isolating compounds and their salts are commonplace in the art.

[0222] In some embodiments, the CDK2 inhibitor may be an isotope-labeled compound or a pharmaceutically acceptable salt thereof. An “isotope-labeled” or “radiolabeled” compound is a compound of the Disclosure in which one or more atoms are replaced or substituted by atoms having an atomic mass or mass number different from that normally found in nature (i.e., naturally occurring atomic mass or mass number). Suitable radionuclides that may be incorporated into the compounds of the Disclosure include: 2 H (deuterium is also written as D), 3 H (Tritium is also written as T), 11 C, 13 C, 14 C, 13 N, 15 N, 15 O,17 O, 18 O, 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 I is one example, but is not limited to these. For example, one or more hydrogen atoms in the compounds of this disclosure may be replaced by deuterium atoms (e.g., C 1~6 One or more hydrogen atoms in an alkyl group may be substituted with deuterium atoms (for example, -CH3 may be substituted with -CD3).

[0223] One or more constituent atoms of the compounds described herein may be replaced or substituted with isotopes of atoms of natural or unnatural abundance. In some embodiments, the compounds contain at least one deuterium atom. In some embodiments, the compounds contain two or more deuterium atoms. In some embodiments, the compounds contain one to two, one to three, one to four, one to five, or one to six deuterium atoms. In some embodiments, all hydrogen atoms in the compound may be replaced or substituted with deuterium atoms.

[0224] Methods for synthesizing the inclusion of isotopes in organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971), The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765, The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopic-labeled compounds can be used in a variety of studies, including NMR spectroscopy, metabolic experiments, and / or assays.

[0225] Substitution with heavier isotopes, such as deuterium, may be preferable in some situations because it can lead to certain therapeutic benefits resulting from greater metabolic stability, such as an increased in vivo half-life or a reduced required dose (see, for example, A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210 and R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may result in one or more therapeutic benefits.

[0226] Therefore, in some embodiments, the CDK2 inhibitor is a compound in which one or more hydrogen atoms in the compound are replaced by deuterium atoms, or a pharmaceutically acceptable salt thereof.

[0227] The term “compound,” as used herein, means all stereoisomers, geometric isomers, tautomers, and isotopes of the illustrated structure. A compound identified by name or structure as one particular tautomer is intended to include other tautomers unless otherwise specified.

[0228] The term "pharmaceutically acceptable" is used herein to mean a compound, substance, composition, and / or dosage form that is suitable for use in contact with human and animal tissues within the bounds of appropriate medical judgment, and that provides a reasonable benefit-to-risk ratio without causing excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0229] This disclosure also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, “pharmaceutically acceptable salt” means a derivative of the disclosed compound in which the parent compound has been modified by converting an existing acidic or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts of this disclosure include, for example, conventional non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of this disclosure can be synthesized from the parent compound containing the base or acid moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or base form of the compound with a stoichiometric amount of a suitable base or acid in water or an organic solvent, or in a mixture thereof, the solvent is generally a non-aqueous medium such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), the entirety of each of which is incorporated herein by reference.

[0230] The term "CDK2 inhibitor" includes any compound that inhibits CDK2, as well as its pharmaceutically acceptable salts, hydrates, solvates, and polymorphs.

[0231] synthesis The compounds of the present invention (including salts thereof) can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, such as those in the following scheme.

[0232] The reactions for preparing the compounds of the present invention may be carried out in suitable solvents that can be readily selected by those skilled in the art of organic synthesis. Suitable solvents may be substantially inactive with the starting materials (reactants), intermediates, or products at the temperature in which the reaction is carried out, for example, in the range from the freezing temperature to the boiling temperature of the solvent. A given reaction may be carried out in one solvent or a mixture of two or more solvents. Depending on the specific reaction step, a solvent suitable for that particular reaction step may be selected by those skilled in the art.

[0233] The preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection, as well as the selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemical properties of protecting groups can be found, for example, in Kocienski, Protecting Groups (Thieme, 2007), Robertson, Protecting Group Chemistry (Oxford University Press, 2000), and Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6. th This is described in Ed. (Wiley, 2007), Peturssion et al., “Protecting Groups in Carbohydrate Chemistry,” J. Chem. Educ., 1997, 74(11), 1297, and Wuts et al., Protective Groups in Organic Synthesis, 4th Ed., (Wiley, 2006).

[0234] The reaction may be monitored according to any suitable method known in the art. For example, product formation may be monitored by spectroscopic means, e.g., nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) It can be monitored by infrared spectroscopy, spectrophotometric methods (e.g., ultraviolet-visible), mass spectrometry, or by chromatographic methods such as high-performance liquid chromatography (HPLC) or thin-layer chromatography (TLC).

[0235] The following scheme provides general guidelines relating to the preparation of the compounds of the present invention. Those skilled in the art will understand that the preparations shown in the scheme can be modified or optimized using general knowledge of organic chemistry to prepare various compounds of the present invention.

[0236] Compounds of formula (AI) can be prepared, for example, using processes such as those shown in schemes 1 and 2 below.

[0237] Compounds of formula (AI) can be prepared from an intermediate of general formula (A). Intermediate (A) can be prepared as shown in Scheme 1. Scheme 1 shows that when the diacid of formula 1-1 is converted to a suitable diester, e.g., methyl or ethyl ester, compound 1-2 can be obtained, and when this is formylated with a suitable reagent (e.g., methyl or ethyl formate), compound 1-3 can be obtained. Compounds of formula 1-4 can be obtained by reacting compounds of formula 1-3 with a suitable guanidine source, e.g., guanidine carbonate or guanidine hydrochloride. Finally, structures of general formula (A) can be obtained by reacting compounds of formula 1-4 with a suitable chlorinating reagent, e.g., phosphorus oxychloride. [ka]

[0238] Intermediates of general formula A can be converted to compounds of formula (I), as shown in scheme 2, having various substituents on R1. Compounds of formula (A) can be reacted with a suitable R2 substituent using various methods (e.g., reductive amination with an aldehyde or ketone, Buchwald-Hartwig amination, copper-catalyzed amination, amide bond formation, etc.) to yield compounds of formula 2-2. When the chloro group of a compound of formula 2-2 reacts with a suitable amine under Buchwald-Hartwig amination conditions, compounds of formula (I) can be yielded. [ka]

[0239] Compounds of formula (BI) can be prepared by various methods depending on the position where the change is desired. For example, a compound of formula (BI) having a change in ring A can be prepared as shown in Scheme 3. In the process illustrated in Scheme 3, the compound of formula 1-2 is provided by selective substitution of the chloro group of trihalopyrimidine 1-1 with the desired amine. Intermediate 1-2 can be reacted by selective Negishi cross-coupling (CCR) with a suitable palladium precatalyst / ligand combination (e.g., Pd2(dba)3 and QPhos or XPhos) to produce intermediate 1-3. Intermediate 1-3 can then be reacted by base-promoted cyclization to provide the compound of formula 1-4. Subsequently, when the desired substitution α is introduced to the amide of intermediate 1-4 (e.g., by sequential alkylation or Pd-catalyzed arylation), the compound of formula 1-5 can be provided. Alternatively, the compounds of formulas 1-5 are provided by reaction with a biselectrophile (e.g., 1,2-dibromoethane) under standard alkylation conditions, where R2 and R3 combine to form a ring, thus providing the compounds of formulas 1-5. Finally, the compound of formula (BI) is provided by Buchwald-Hartwig amination with a suitable amine. [ka]

[0240] R 1 Compounds of formula (BI) having different groups can be formed as shown in scheme 4. Thus, as described above, the R of compound 2-1 2 and R 3 The introduction of compound 2-2 provides which can undergo selective oxidation of sulfur (e.g., using m-CPBA) to provide intermediate 2-3. The intermediate 2-3 with the resulting sulfone is selectively SN with a suitable N-formylamine. ArThe reaction provides intermediates 2-4. Finally, the reaction of intermediates 2-4 with a suitable amine provides a compound of general formula (BI). This coupling can be carried out in one of two ways. Firstly, a tandem Buchwald-Hartwig amination and cyclization catalyzed by a suitable pre-formed catalyst (e.g., RuPhos second-generation pre-catalyst or XantPhos second-generation pre-catalyst) may be used. Alternatively, the two-step protocol may be SN-1 with a suitable acidic (TFA) or basic (Hünig base) catalyst and a suitable polar solvent (i.e., 1,1,1-trifluoroethanol or 1-butanol). Ar The reaction consists of a reaction followed by a cyclization induced by a suitable base (i.e., sodium hydride). [ka]

[0241] Treatment method The methods disclosed herein enable the assessment of whether human subjects with, suspected of having, or at risk of developing a CDK2-related disease or disorder may respond to a CDK2 inhibitor (e.g., whether they have a significant improvement in the disease, as demonstrated by remission / recovery, or whether CDK2 may be inhibited). Human subjects with, suspected of having, or at risk of developing a CDK2-related disease or disorder who may respond to a CDK2 inhibitor may be administered a CDK2 inhibitor. Conversely, human subjects with, suspected of having, or at risk of developing a CDK2-related disease or disorder who are unlikely to respond to a CDK2 inhibitor may be administered additional therapies suitable for treating the disease or disorder.

[0242] The method described herein also makes it possible to stratify human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder into groups of subjects who are likely to benefit from treatment including CDK2 inhibitors and groups who are unlikely to benefit. The ability to select such human subjects from a pool of human subjects with CDK2-related diseases or disorders for which treatment with CDK2 inhibitors is being considered is beneficial for providing effective treatment to the subjects.

[0243] In one embodiment, a human subject treated with a CDK2 inhibitor has, is suspected of having, or is at risk of developing a CDK2-related disease or disorder. In a particular embodiment, a human subject treated with a CDK2 inhibitor has, is suspected of having, or is at risk of developing cancer.

[0244] Human subjects with a disease or disorder associated with CDK2 may be administered an effective dose of a CDK2 inhibitor if they are likely to respond to one or more of the markers listed above (e.g., biomarkers or pharmacodynamic markers, e.g., CCNE1, p16, and Rb phosphorylation). The effective dose of the CDK2 inhibitor may be appropriately determined by a healthcare professional, taking into account, for example, the patient's characteristics (age, sex, weight, race, etc.), disease progression, and prior exposure to the drug. If a human subject is unlikely to respond to a CDK2 inhibitor, they may be administered a therapy that does not include a CDK2 inhibitor.

[0245] After stratifying or selecting human subjects based on whether they are more or less likely to respond to CDK inhibitors, a medical practitioner (e.g., a doctor) can administer appropriate treatment to the human subjects. Methods for administering CDK2 inhibitors are known in the art.

[0246] If a human subject with a CDK2-related disease or disorder, and who is expected to respond to a CDK2 inhibitor, has previously received one or more non-CDK2 inhibitor therapies, then a CDK2 inhibitor may replace or enhance the previously or currently administered therapy. For example, treatment with a CDK2 inhibitor may lead to the discontinuation or reduction of one or more non-CDK2 inhibitor therapies, for example, by administering them at lower levels. While a CDK2 inhibitor is administered, the administration of previous therapies may be maintained. In some embodiments, previous therapies may be maintained until the level of the CDK2 inhibitor reaches a level sufficient to produce a therapeutic effect.

[0247] In certain embodiments, methods are provided herein for treating human subjects who have, are suspected of having, or are at risk of developing a disease or disorder associated with CDK2, the methods comprising administering a CDK2 inhibitor to the human subjects, the human subjects having been determined to (i)(a) have a nucleotide sequence encoding a p16 protein comprising the amino acid sequence of SEQ ID NO: 1, (b) have a CDKN2A gene lacking one or more inactivating nucleic acid substitutions and / or deletions, and / or (c) express a p16 protein (e.g., a p16 protein comprising the amino acid sequence of SEQ ID NO: 1), and (ii)(a) have an amplified CCNE1 gene and / or (b) have a higher level of CCNE1 expression in a biological sample obtained from the subjects than the control expression level of CCNE1. In certain embodiments, the biological sample was obtained from a human subject at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months prior to administration of the CDK2 inhibitor. In certain embodiments, the biological sample was obtained from a human subject up to 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months prior to administration of the CDK2 inhibitor. In certain embodiments, the subject was determined to have the gene encoding the p16 protein of Sequence ID No. 1 at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months prior to administration of the CDK2 inhibitor. In certain embodiments, subjects were determined to have the gene encoding the p16 protein of Sequence ID No. 1 up to 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, or 2 months prior to administration of the CDK2 inhibitor. In certain embodiments, the method further includes: (1) After administering a CDK2 inhibitor to the subject, measure the decrease in the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in the biological sample obtained from the subject, compared with the control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, and (2) After measurement, continue administering the CDK2 inhibitor to human subjects.

[0248] In certain embodiments, biological samples obtained from subjects after administration of a CDK2 inhibitor were obtained at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after administration of the CDK2 inhibitor. In certain embodiments, biological samples obtained from subjects after administration of a CDK2 inhibitor were obtained from human subjects up to 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after administration of the CDK2 inhibitor. In certain embodiments, the continuation of step (2) is performed at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months after the measurement in step (1). In certain embodiments, the continuation of step (2) is performed up to 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months after the measurement in step (1).

[0249] In other specific embodiments, methods are provided herein for treating human subjects who have, are suspected of having, or are at risk of developing a CDK2-related disease or disorder, the method comprising (i) identifying in a biological sample obtained from a human subject the presence of (a) a nucleotide sequence encoding the p16 protein including the amino acid sequence of SEQ ID NO: 1, (b) a CDKN2A gene lacking one or more inactivating nucleic acid substitutions, and / or (c) the p16 protein (e.g., the p16 protein including the amino acid sequence of SEQ ID NO: 1), and (ii) identifying in a biological sample obtained from a human subject (a) amplification of the CCNE1 gene and / or (b) an expression level of CCNE1 higher than the control expression level of CCNE1, and administering a CDK2 inhibitor to the human subject. In specific embodiments, the human subject has a CDK2-related disease or disorder. In specific embodiments, the human subject is suspected of having, or is at risk of developing, a CDK2-related disease or disorder. In certain embodiments, administration is performed at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, or at least 2 months after identifying amplification of the CDKN2A gene, p16 protein, and / or CCNE1 gene and / or CCNE1 expression levels higher than the control expression level of CCNE1 in a biological sample obtained from a human subject. In certain embodiments, administration is performed up to 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, or 2 months after identifying in a biological sample obtained from a human subject the presence of a nucleotide sequence encoding the p16 protein containing the amino acid sequence of SEQ ID NO: 1, a CDKN2A gene lacking one or more inactivating nucleic acid substitutions, and / or the p16 protein, and / or amplification of the CCNE1 gene and / or a CCNE1 expression level higher than the control expression level of CCNE1.In certain embodiments, the method further includes measuring the decrease in the level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample obtained from a subject after administration of a CDK2 inhibitor to the subject, compared to a control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, and continuing to administer the CDK2 inhibitor to the human subject after the measurement. In certain embodiments, the biological sample obtained from a subject after administration of a CDK2 inhibitor was obtained from a human subject at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 2 weeks, at least 3 weeks, or at least 4 weeks after administration. In certain embodiments, biological samples obtained from subjects after administration of a CDK2 inhibitor were taken from human subjects at up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, or 4 weeks after administration. In certain embodiments, continued administration was performed at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, or 2 months after measurement. In certain embodiments, continued administration may occur at a maximum of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, or 2 months after measurement.

[0250] In some embodiments, the diseases or disorders associated with CDK2 include N-myc-amplified neuroblastoma cells (see Molenaar, et al., Proc Natl Acad Sci USA 106(31):12968-12973), K-Ras mutant lung cancer (see Hu, S., et al., Mol Cancer Ther, 2015.14(11):p.2576-85), and cancers with FBW7 mutations and CCNE1 overexpression (see Takada, et al., Cancer Res, 2017.77(18):p.4881-4893).

[0251] In some embodiments, the diseases or disorders associated with CDK2 are squamous cell carcinoma of the lung, adenocarcinoma of the lung, adenocarcinoma of the pancreas, invasive breast cancer, carcinosarcoma of the uterus, serous cystadenocarcinoma of the ovary, adenocarcinoma of the stomach, esophageal cancer, urothelial carcinoma of the bladder, mesothelioma, or sarcoma.

[0252] In some embodiments, the disease or disorder associated with CDK2 is lung adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, or gastric adenocarcinoma.

[0253] In some embodiments, the disease or disorder associated with CDK2 is adenocarcinoma, carcinoma, or cystadenoma.

[0254] In some embodiments, the disease or disorder associated with CDK2 is uterine cancer, ovarian cancer, gastric cancer, esophageal cancer, lung cancer, bladder cancer, pancreatic cancer, or breast cancer.

[0255] In some embodiments, the disease or disorder associated with CDK2 is cancer.

[0256] In some embodiments, the cancer is characterized by amplification or overexpression of CCNE1. In some embodiments, the cancer is ovarian cancer or breast cancer characterized by amplification or overexpression of CCNE1.

[0257] In some embodiments, breast cancer is resistant to chemotherapy or radiotherapy, endocrine-resistant, trastuzumab-resistant, or primary or acquired resistance to CDK4 / 6 inhibition. In some embodiments, breast cancer is advanced or metastatic breast cancer.

[0258] Examples of cancers treatable using the methods of this disclosure include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, such as acute myeloid leukemia, chronic bone cancer. This includes, but is not limited to, myelin leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors in children, lymphocytic lymphoma, bladder cancer, kidney or urethral cancer, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers, such as asbestos-induced cancer, and combinations of such cancers. The methods of this disclosure are also useful for treating metastatic cancers.

[0259] In some embodiments, cancers treatable by the methods of this disclosure include melanoma (e.g., metastatic malignant melanoma, BRAF and HSP90 inhibitor-resistant melanoma), renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory adenocarcinoma of the prostate), breast cancer, colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), head and neck squamous cell carcinoma, urothelial carcinoma (e.g., bladder cancer), and high-frequency microsatellite instability (MSI). high Examples include cancers accompanied by ). Furthermore, this disclosure includes refractory or recurrent malignancies whose growth may be inhibited using the methods of this disclosure.

[0260] In some embodiments, cancers treatable using the methods of this disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., lymphoma, leukemia, e.g., acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin lymphoma (including follicular lymphoma, including relapsed or refractory NHL and relapsed follicular), Hodgkin lymphoma, or multiple myeloma, etc.), and combinations of such cancers.

[0261] In some embodiments, cancers treatable using the methods of the present disclosure include, but are not limited to, cholangiocarcinoma, bile duct cancer, triple-negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing's sarcoma, brain cancer, brain tumors, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, islet cell carcinoma, oral cancer, mouth cancer, pharyngeal cancer, laryngeal cancer, lip cancer, mesothelioma, cervical cancer, nasal cavity cancer, ocular cancer, melanoma, pelvic cancer, rectal cancer, renal cell carcinoma, salivary gland cancer, paranasal sinus cancer, spinal cancer, tongue cancer, tubular cancer, urethral cancer, and ureteral cancer.

[0262] In some embodiments, the methods disclosed herein may be used to treat sickle cell disease and sickle cell anemia.

[0263] In some embodiments, diseases and indications treatable using the methods of this disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, urogenital cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

[0264] Exemplary blood cancers include lymphomas and leukemias, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular lymphoma), Hodgkin lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocytosis (ET)), myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), and multiple myeloma (MM).

[0265] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdosarcoma, fibroma, lipoma, hamartoma, and teratoma.

[0266] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), bronchogenic lung cancer, squamous cell carcinoma, anaplastic small cell carcinoma, anaplastic large cell carcinoma, adenocarcinoma, alveolar (bronchiolar) carcinoma, bronchial adenoma, chondrotoxic hamartoma, and mesothelioma.

[0267] Examples of gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine cancer (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma), and colorectal cancer.

[0268] Examples of urogenital tract cancers include cancers of the kidney (adenocarcinoma, Wilms' tumor [nephroblastoma]), cancers of the bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), cancers of the prostate (adenocarcinoma, sarcoma), and cancers of the testes (seminocarcinoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma).

[0269] Examples of liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0270] Examples of bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulosarcoma), multiple myeloma, malignant giant cell chordoma, osteochondroma (osteochondrial exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor.

[0271] Exemplary neurological cancers include cancers of the skull (osteoma, hemangioma, granuloma, xanthomas, osteoosteitis), cancers of the meninges (meningioma, meningiosarcoma, gliomas), cancers of the brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal glandoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), and cancers of the spinal cord (neurofibroma, meningioma, glioma, sarcoma), as well as neuroblastoma and Lhermitt-Dukuro disease.

[0272] Examples of gynecological cancers include cancer of the uterus (endometrial cancer), cancer of the cervix (cervical cancer, preneoplastic cervical dysplasia), cancer of the ovaries (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiable cancer), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), cancer of the vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), cancer of the vagina (clear cell carcinoma, squamous cell carcinoma, staphylosarcoma (embryonic rhabdomyosarcoma)), and cancer of the fallopian tubes (carcinoma).

[0273] Examples of skin cancers include melanoma, basal cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, hemangioma, dermatofibroma, and keloid. In some embodiments, diseases and indications that can be treated with the compounds of this disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple-negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, cholangiocarcinoma, esophageal cancer, and urothelial carcinoma.

[0274] Combination therapy Human subjects treated with CDK2 inhibitors according to the methods described herein may be treated in combination with one or more additional compositions or therapies effective in treating CDK2-related diseases or disorders. In some embodiments, CDK2 inhibitors are administered or used in combination with BCL2 inhibitors or CDK4 / 6 inhibitors.

[0275] I. Cancer Therapy Cancer cell proliferation and survival can be affected by dysfunction of multiple signaling pathways. Therefore, it is useful to treat such conditions by combining different enzyme / protein / receptor inhibitors that exhibit different selectivity in targets, such as those that modulate the activity of the target. Targeting two or more signaling pathways (or two or more biomolecules involved in a given signaling pathway) may reduce the potential for drug resistance to develop in the cell population and / or reduce the toxicity of the treatment.

[0276] One or more additional pharmaceuticals, such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, cancer immunotherapeutic agents, metabolic enzyme inhibitors, chemokine receptor inhibitors, and phosphatase inhibitors, as well as targeted therapies, such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF, FAK, and CDK4 / 6 kinase inhibitors, such as those described in WO2006 / 056399, may be used in combination with the compounds of the Disclosure to treat CDK2-related diseases, disorders, or conditions. Other agents, such as therapeutic antibodies, may be used in combination with the compounds of the Disclosure to treat CDK2-related diseases, disorders, or conditions. One or more additional pharmaceuticals may be administered to the patient simultaneously or sequentially.

[0277] In some embodiments, CDK2 inhibitors are administered or used in combination with BCL2 inhibitors or CDK4 / 6 inhibitors.

[0278] Compounds such as those disclosed herein may be used in combination with one or more other enzyme / protein / receptor inhibitor therapies to treat diseases such as cancer and other diseases or disorders described herein. Examples of diseases and indications treatable with combination therapy are those described herein. Examples of cancer include solid and non-solid tumors, e.g., humoral tumors, hematological malignancies. Examples of infections include viral infections, bacterial infections, fungal infections, or parasitic infections. For example, the compounds disclosed herein may be used in combination with one or more inhibitors of the following kinases to treat cancer: Akt1, Akt2, Akt3, BCL2, CDK4 / 6, TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFαR, PDGFβR, PI3K (alpha, beta, gamma, delta, and multiple or selective), C SF1R, KIT, FLK-II, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, PARP, Ron, Sea, TRKA, TRKB, TRKC, TAM kinase (Axl, Mer, Tyro3), FLT3, VEGFR / Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK, and B-Raf. In some embodiments, the compounds of the present disclosure may be combined with one or more of the following inhibitors to treat cancer or infection. Non-limiting examples of inhibitors that may be combined with the compounds of this disclosure to treat cancer and infectious diseases include FGFR inhibitors (FGFR1, FGFR2, FGFR3, or FGFR4, e.g., pemigatinib (INCB54828), or INCB62079), EGFR inhibitors (also known as ErB-1 or HER-1, e.g., erlotinib, gefitinib, vandetanib, orsimertinib, cetuximab, necitumumab, or panitumumab), VEGFR inhibitors or pathway blockers (e.g., bevacizumab, pazopanib, sunitinib, sorafenib,Axitinib, regorafenib, ponatinib, cabozantinib, vandetanib, ramucirumab, lenvatinib, ziv-aflibercept), PARP inhibitors (e.g., olaparib, rucaparib, veliparib or niraparib), JAK inhibitors (JAK1 and / or JAK2, e.g., ruxolitinib or baricitinib, or JAK1 inhibitors, e.g., itacitinib (INCB039110), INCB052793, or INCB054707), seitasi Tinib (INCB39110), IDO inhibitors (e.g., epacadostat, NLG919, or BMS-986205, MK7162), LSD1 inhibitors (e.g., GSK2979552, INCB59872, or INCB60003), TDO inhibitors, PI3K-delta inhibitors (e.g., pulsacricib (INCB50465) or INCB50797), PI3K-gamma inhibitors, such as PI3K-gamma selective inhibitors, and Pim inhibitors (e.g., INCB 53914), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl, and Mer, e.g., INCB081776), adenosine receptor antagonists (e.g., A2a / A2b receptor antagonists), HPK1 inhibitors, chemokine receptor inhibitors (e.g., CCR2 or CCR5 inhibitors), SHP1 / 2 phosphatase inhibitors, histone deacetylase inhibitors (HDACs), e.g., HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors Examples include bromo-domain inhibitors, bromo- and extra-terminal family member inhibitors (e.g., bromo-domain inhibitors or BET inhibitors, e.g., INCB54329 and INCB57643), TAM receptor tyrosine kinase inhibitors (Tyro-3, Axl, and Mer, e.g., INCB81776), c-MET inhibitors (e.g., capmatinib), anti-CD19 antibodies (e.g., tafacitamab), ALK2 inhibitors (e.g., INCB00928), or combinations thereof.

[0279] In some embodiments, the compounds or salts described herein are administered together with a PI3Kδ inhibitor. In some embodiments, the compounds or salts described herein are administered together with a JAK inhibitor. In some embodiments, the compounds or salts described herein are administered together with a JAK1 or JAK2 inhibitor (e.g., baricitinib or ruxolitinib). In some embodiments, the compounds or salts described herein are administered together with a JAK1 inhibitor. In some embodiments, the compounds or salts described herein are administered together with a JAK1 inhibitor that is more selective than JAK2 inhibitors.

[0280] Examples of antibodies for use in combination therapy include, but are not limited to, trastuzumab (e.g., anti-HER2), ranibizumab (e.g., anti-VEGF-A), bevacizumab (AVASTIN®, e.g., anti-VEGF), panitumumab (e.g., anti-EGFR), cetuximab (e.g., anti-EGFR), rituxan (e.g., anti-CD20), and antibodies targeting c-MET.

[0281] One or more of the following drugs may be used in combination with the compounds of this disclosure, and are presented as a non-limiting list: the cell proliferation inhibitors cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptosar, topotecan, paclitaxel, docetaxel, epotilon, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH66336, R115777, L778,123, BMS 214662, IRESSA (trademark) (gefitinib), TARCEVA (trademark) (erlotinib), antibody against EGFR, intron, ara-C, Adriamycin, cytoxan, gemcitabine, uracil mustard, chlormetine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxlidine N, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovorin, ELOXATIN (trademark) (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17. alpha.- Ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianicene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisol, navelbine, anastrazole, letrozole, capecitabine, reloxafine, droxifen, hexamethylmelamine, avastin, HERCEPTIN (trademark) (trastuzumab), BEXXAR (trademark) (Toshitumomab), VELCADE (bortezomib), ZEVALIN (ibritumomab tiuxetan), TRISENOX (arsenic trioxide), XELODA (capecitabine), vinorelbine, porfimer, ERBITUX (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, letrozole, fulvestrant, exemestate Ifosfamide, rituximab, C225 (cetuximab), Campath (aremutuzumab), clopharabine, cladribine, aphidicolin, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP, and MDL-101,731.

[0282] The compounds of this disclosure may also be used in combination with other methods of treating cancer, such as chemotherapy, radiotherapy, tumor-targeted therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapies include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, bispecific or multispecific antibodies, antibody-drug conjugates, adoptive T cell transfer, Toll receptor agonists, RIG-I agonists, oncolytic virus therapy, and immunomodulatory small molecules, such as thalidomide or JAK1 / 2 inhibitors, PI3Kδ inhibitors, etc. The compounds may be administered in combination with one or more anticancer drugs, such as chemotherapeutic agents. Examples of chemotherapy drugs include avalerix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, busulfan for intravenous injection, busulfan for oral use, carsterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin difutitox, dexrazoxane, docetaxel, doxorubicin, and dromostanol propionate. Lon, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alpha-2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisol, lomustine, mechloretamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane,Mitoxantrone, Nandrolone fenpropionate, Nelarabine, Nofetumomab, Oxaliplatin, Paclitaxel, Pamidronate, Panitumumab, Pegasparagase, Pegfilgrastim, Pemetrexed disodium, Pentostatin, Pipobroman, Plicamycin, Procarbazine, Quinacrine, Rasburicase, Rituximab, Ruxolitinib, Sorafenib, St. Examples include leptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tocitumomab, trastuzumab, tretinoin, uracil mustard, barrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronate.

[0283] Examples of additional chemotherapy drugs include proteasome inhibitors (e.g., bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, and carmustine.

[0284] Examples of steroids include corticosteroids, such as dexamethasone or prednisone.

[0285] Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC®), nilotinib, dasatinib, bosutinib, and ponatinib, as well as pharmaceutically acceptable salts. Other exemplary and suitable Bcr-Abl inhibitors include compounds of the genera and species disclosed in U.S. Patent No. 5,521,184, WO04 / 005281, and U.S. Provisional Patent Application No. 60 / 578,491, and their pharmaceutically acceptable salts.

[0286] Exemplary and suitable Flt-3 inhibitors include midostaurin, restaurtinib, linifanib, sunitinib, sunitinib maleate, sorafenib, quizartinib, clenolanib, pacritinib, tanzutinib, PLX3397 and ASP2215, and pharmaceutically acceptable salts thereof. Other exemplary and suitable Flt-3 inhibitors include compounds such as those disclosed in WO03 / 037347, WO03 / 099771 and WO04 / 046120, and pharmaceutically acceptable salts thereof.

[0287] Exemplary and suitable RAF inhibitors include dabrafenib, sorafenib, and vemurafenib, and their pharmaceutically acceptable salts. Other exemplary and suitable RAF inhibitors include compounds such as those disclosed in WO00 / 09495 and WO05 / 028444, and their pharmaceutically acceptable salts.

[0288] Exemplary and preferred FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520, and GSK2256098, as well as pharmaceutically acceptable salts thereof. Other exemplary and preferred FAK inhibitors include compounds such as those disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, as well as pharmaceutically acceptable salts thereof.

[0289] Exemplary and suitable CDK4 / 6 inhibitors include palbociclib, ribociclib, trilaciclib, rerocyclib, and abemaciclib, as well as pharmaceutically acceptable salts thereof. Other exemplary and suitable CDK4 / 6 inhibitors include compounds such as those disclosed in WO09 / 085185, WO12 / 129344, WO11 / 101409, WO03 / 062236, WO10 / 075074, and WO12 / 061156, and pharmaceutically acceptable salts thereof.

[0290] In some embodiments, the compounds of the present disclosure may be used in combination with one or more other kinase inhibitors, including imatinib, particularly to treat patients who are resistant to imatinib or other kinase inhibitors.

[0291] In some embodiments, the compounds of this disclosure may be used in combination with chemotherapeutic agents in the treatment of cancer, and may improve the treatment response compared to the response with the chemotherapeutic agent alone without causing an exacerbation of its toxic effects. In some embodiments, the compounds of this disclosure may be used in combination with chemotherapeutic agents provided herein. For example, additional pharmaceuticals used in the treatment of multiple myeloma may include, but are not limited to, melphalan, melphalan and prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Further additional agents used in the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF, and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulator. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulator is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable outcomes when the CDK2 inhibitors of this disclosure are combined with additional agents.

[0292] The drugs may be combined with the compound in a single or sequential dosage form, or the drugs may be administered simultaneously or sequentially in separate dosage forms.

[0293] The compounds of this disclosure may be used in combination with one or more other inhibitors or therapies for treating infectious diseases. Examples of infectious diseases include viral infections, bacterial infections, fungal infections, or parasitic infections.

[0294] In some embodiments, a corticosteroid such as dexamethasone is administered to the patient in combination with the compounds of the present disclosure, and the dexamethasone is administered intermittently rather than continuously.

[0295] Compounds described herein, compounds listed in any of the claims, or salts thereof may be combined with other immunogenic agents, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), cells, and cells transfected with genes encoding immunostimulatory cytokines. Non-limiting examples of tumor vaccines that may be used include melanoma antigen peptides, such as gp100, MAGE antigen, Trp-2, MARTI, and / or tyrosinase peptides, or tumor cells transfected to express the cytokine GM-CSF.

[0296] Compounds described herein, compounds listed in any of the claims, or salts thereof may be used in combination with vaccination protocols intended for the treatment of cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins derived from viruses involved in human cancer, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, the compounds of this disclosure may be used in combination with tumor-specific antigens, such as heat shock proteins isolated from tumor tissue itself. In some embodiments, compounds described herein, compounds listed in any of the claims, or salts thereof may be used in combination with dendritic cell immunity to activate a potent antitumor response.

[0297] The compounds of this disclosure may be used in combination with bispecific macrocyclic peptides that target tumor cells with Fe-alpha or Fe-gamma receptor-expressing effector cells. The compounds of this disclosure may also be combined with macrocyclic peptides that activate the host immune response.

[0298] In some further embodiments, the compounds of the present disclosure may be administered to a patient in combination with other therapeutic agents before, during, and / or after bone marrow transplantation or stem cell transplantation. The compounds of the present disclosure may be used in combination with bone marrow transplantation to treat various tumors of hematopoietic origin.

[0299] Compounds described herein, compounds listed in any of the claims, or salts thereof may be used in combination with vaccines to stimulate an immune response to pathogens, toxins, and autoantigens. Examples of pathogens for which this therapeutic approach may be particularly useful include pathogens for which no effective vaccine currently exists, or for which conventional vaccines are not entirely effective. These include, but are not limited to, HIV, hepatitis (types A, B, and C), influenza, herpes, giardia, malaria, leishmania, Staphylococcus aureus, and Pseudomonas aeruginosa.

[0300] Viruses that cause infectious diseases treatable by the methods of this disclosure include, but are not limited to, human papillomavirus, influenza, hepatitis A, B, C, or D viruses, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus, measles virus, herpesvirus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus), flavivirus, echovirus, rhinovirus, coxsackievirus, cornovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus, and arbovirus encephalitis virus.

[0301] Pathogenic bacteria that cause infections treatable by the methods of this disclosure include, but are not limited to, Chlamydia, Rickettsia, Mycobacterium, Staphylococcus, Streptococcus, Pneumococcus, Meningococcus and Conococcus, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacillus, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease.

[0302] Pathogenic fungi that cause infections treatable by the methods disclosed herein include, but are not limited to, Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Mucorales (mucor, humicaria, rhinoplakia), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum.

[0303] Pathogenic parasites that cause infections treatable by the methods disclosed herein include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba species, Giardia lambia, Cryptosporidium species, Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

[0304] When two or more medications are administered to a patient, they may be administered simultaneously, separately, sequentially, or in combination (for example, in the case of three or more medications).

[0305] Methods for safely and effectively administering many of these chemotherapeutic agents are known to those skilled in the art. In addition, their administrations are documented in standard literature. For example, the administrations of many chemotherapeutic agents are described in the "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), and its disclosures are incorporated herein by reference as they are described in their entirety.

[0306] II. Immunotherapy Checkpoint Therapy The compounds of this disclosure may be used in combination with one or more immune checkpoint inhibitors to treat diseases such as cancer or infections. Exemplary immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CBL-B, CD20, CD28, CD40, CD70, CD122, CD96, CD73, CD47, CDK2, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, arginase, HPK1, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TLR (TLR7 / 8), TIGIT, CD112R, VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulant checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT, and VISTA. In some embodiments, the compounds provided herein may be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFR beta inhibitors.

[0307] In some embodiments, the compounds provided herein may be used in combination with one or more agonists of immune checkpoint molecules, such as OX40, CD27, GITR, and CD137 (also known as 4-1BB).

[0308] In some embodiments, the inhibitor of the immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0309] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PD-1 or PD-L1, for example, an anti-PD-1 or anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 or anti-PD-L1 antibody is nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, semiprimab, atezolizumab, avelumab, tislerizumab, spartalizumab (PDR001), cetrelimab (JNJ-63723283), tripalimab (JS001), camrelizumab (SHR-1210), cintilimab (IBI308), AB122 (GLS-010), AMP-224, AMP-514 / MEDI- These are 0680, BMS936559, JTX-4014, BGB-108, SHR-1210, MEDI4736, FAZ053, BCD-100, KN035, CS1001, BAT1306, LZM009, AK105, HLX10, SHR-1316, CBT-502(TQB2450), A167(KL-A167), STI-A101(ZKAB001), CK-301, BGB-A333, MSB-2311, HLX20, TSR-042, or LY3300054.In some embodiments, the inhibitor of PD-1 or PD-L1 is U.S. Patent No. 7,488,802, No. 7,943,743, No. 8,008,449, No. 8,168,757, No. 8,217,149, or No. 10,308,644, U.S. Patent Publication No. 2017 / 0145025, No. 2017 / 0174671, No. 2017 / 0174679, No. 20 17 / 0320875, 2017 / 0342060, 2017 / 0362253, 2018 / 0016260, 2018 / 0057486, 2018 / 017 7784, 2018 / 0177870, 2018 / 0179179, 2018 / 0179201, 2018 / 0179202, 2018 / 0273519, Nos. 2019 / 0040082, 2019 / 0062345, 2019 / 0071439, 2019 / 0127467, 2019 / 0144439, 2019 / 0202824, 2019 / 0225601, 2019 / 0300524, or 2019 / 0345170, or PCT Publication Nos. WO03 / 042402, WO These are disclosed in publications 2008 / 156712, WO2010 / 089411, WO2010 / 036959, WO2011 / 066342, WO2011 / 159877, WO2011 / 082400, WO2011 / 161699, or WO2019 / 246110, each of which is incorporated herein by reference in its entirety. In some embodiments, the PD-L1 inhibitor is INCB086550.

[0310] In some embodiments, the antibody is an anti-PD-1 antibody, for example, an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, semiprimab, spartalizumab, camrelizumab, cetrelimab, tripalimab, scintirimab, AB122, AMP-224, JTX-4014, BGB-108, BCD-100, BAT1306, LZM009, AK105, HLX10, or TSR-042. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, semiprimab, spartalizumab, camrelizumab, cetrelimab, tripalimab, or scintirimab. In some embodiments, the anti-PD-1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the anti-PD-1 antibody is cemiprimab. In some embodiments, the anti-PD-1 antibody is spartalizumab. In some embodiments, the anti-PD-1 antibody is camrelizumab. In some embodiments, the anti-PD-1 antibody is cetrelimab. In some embodiments, the anti-PD-1 antibody is tripalimab. In some embodiments, the anti-PD-1 antibody is cintilimab. In some embodiments, the anti-PD-1 antibody is AB122. In some embodiments, the anti-PD-1 antibody is AMP-224. In some embodiments, the anti-PD-1 antibody is JTX-4014. In some embodiments, the anti-PD-1 antibody is BGB-108. In some embodiments, the anti-PD-1 antibody is BCD-100. In some embodiments, the anti-PD-1 antibody is BAT1306. In some embodiments, the anti-PD-1 antibody is LZM009. In some embodiments, the anti-PD-1 antibody is AK105. In some embodiments, the anti-PD-1 antibody is HLX10. In some embodiments, the anti-PD-1 antibody is TSR-042. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012 (INCMGA0012, retifanlimab). In some embodiments, the anti-PD-1 antibody is SHR-1210.Other anticancer drugs (multiple options are possible) include antibody therapies, such as 4-1BB (e.g., urelumab, utomirumab).

[0311] In some embodiments, the inhibitor of the immune checkpoint molecule is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is atezolizumab, avelumab, durvalumab, tislerizumab, BMS-935559, MEDI4736, atezolizumab (also known as MPDL3280A, RG7446), avelumab (MSB0010718C), FAZ053, KN035, CS1001, SHR-1316, CBT-502, A167, STI-A101, CK-301, BGB-A333, MSB-2311, HLX20, or LY3300054. In some embodiments, the anti-PD-L1 antibody is atezolizumab, avelumab, durvalumab, or tislerizumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab. In some embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is durvalumab. In some embodiments, the anti-PD-L1 antibody is tislerizumab. In some embodiments, the anti-PD-L1 antibody is BMS-935559. In some embodiments, the anti-PD-L1 antibody is MEDI4736. In some embodiments, the anti-PD-L1 antibody is FAZ053. In some embodiments, the anti-PD-L1 antibody is KN035. In some embodiments, the anti-PD-L1 antibody is CS1001. In some embodiments, the anti-PD-L1 antibody is SHR-1316. In some embodiments, the anti-PD-L1 antibody is CBT-502. In some embodiments, the anti-PD-L1 antibody is A167. In some embodiments, the anti-PD-L1 antibody is STI-A101. In some embodiments, the anti-PD-L1 antibody is CK-301. In some embodiments, the anti-PD-L1 antibody is BGB-A333. In some embodiments, the anti-PD-L1 antibody is MSB-2311. In some embodiments, the anti-PD-L1 antibody is HLX20. In some embodiments, the anti-PD-L1 antibody is LY3300054.

[0312] In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds to PD-L1, or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds to and internalizes PD-L1, or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of an immune checkpoint molecule is a compound selected from those described in US2018 / 0179201, US2018 / 0179197, US2018 / 0179179, US2018 / 0179202, US2018 / 0177784, US2018 / 0177870, U.S. Patent Application No. 16 / 369,654 (filed March 29, 2019), and U.S. Provisional Patent Application No. 62 / 688,164, or a pharmaceutically acceptable salt thereof, each of which is incorporated herein by reference in whole.

[0313] In some embodiments, the inhibitors of immune checkpoint molecules are inhibitors of KIR, TIGIT, LAIR1, CD160, 2B4, and TGFR beta.

[0314] In some embodiments, the inhibitor is MCLA-145.

[0315] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

[0316] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of LAG3, for example, an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, INCAGN2385, or eftyragimod alfa (IMP321).

[0317] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is oleculumab.

[0318] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIGIT. In some embodiments, the inhibitor of TIGIT is OMP-31M32.

[0319] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of VISTA. In some embodiments, the inhibitor of VISTA is JNJ-61610588 or CA-170.

[0320] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of B7-H3. In some embodiments, the inhibitor of B7-H3 is enobrituzumab, MGD009, or 8H9.

[0321] In some embodiments, the inhibitor of the immune checkpoint molecule is a KIR inhibitor. In some embodiments, the KIR inhibitor is lirilumab or IPH4102.

[0322] In some embodiments, the inhibitor of the immune checkpoint molecule is an A2aR inhibitor. In some embodiments, the A2aR inhibitor is CPI-444.

[0323] In some embodiments, the inhibitor of the immune checkpoint molecule is a TGF-beta inhibitor. In some embodiments, the TGF-beta inhibitor is travedersen, garcertinib, or M7824.

[0324] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of PI3K-gamma. In some embodiments, the inhibitor of PI3K-gamma is IPI-549.

[0325] In some embodiments, the inhibitor of the immune checkpoint molecule is a CD47 inhibitor. In some embodiments, the CD47 inhibitor is Hu5F9-G4 or TTI-621.

[0326] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is MEDI9447.

[0327] In some embodiments, the inhibitor of the immune checkpoint molecule is a CD70 inhibitor. In some embodiments, the CD70 inhibitor is xatuzumab or BMS-936561.

[0328] In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIM3, for example, an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

[0329] In some embodiments, the inhibitor of the immune checkpoint molecule is a CD20 inhibitor, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0330] In some embodiments, the agonists of immune checkpoint molecules are agonists of OX40, CD27, CD28, GITR, ICOS, CD40, TLR7 / 8, and CD137 (also known as 4-1BB).

[0331] In some embodiments, the agonist of CD137 is urelumab. In some embodiments, the agonist of CD137 is utomirumab.

[0332] In some embodiments, the immune checkpoint molecule agonist is a GITR inhibitor. In some embodiments, the GITR agonist is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, MEDI1873, or MEDI6469.

[0333] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of OX40, such as an OX40 agonist antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is INCAGN01949, MEDI0562 (tavolimab), MOXR-0916, PF-04518600, GSK3174998, BMS-986178, or 9B12. In some embodiments, the OX40L fusion protein is MEDI6383.

[0334] In some embodiments, the agonist of the immune checkpoint molecule is a CD40 agonist. In some embodiments, the CD40 agonist is CP-870893, ADC-1013, CDX-1140, SEA-CD40, RO7009789, JNJ-64457107, APX-005M, or Chi Lob 7 / 4.

[0335] In some embodiments, the immune checkpoint molecule agonist is an ICOS agonist. In some embodiments, the ICOS agonist is GSK-3359609, JTX-2011, or MEDI-570.

[0336] In some embodiments, the agonist of the immune checkpoint molecule is a CD28 agonist. In some embodiments, the CD28 agonist is ceralizumab.

[0337] In some embodiments, the agonist of the immune checkpoint molecule is a CD27 agonist. In some embodiments, the CD27 agonist is valrilumab.

[0338] In some embodiments, the immune checkpoint molecule agonist is a TLR7 / 8 agonist. In some embodiments, the TLR7 / 8 agonist is MEDI9197.

[0339] The compounds of this disclosure may be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3, or the TGFβ receptor. In some embodiments, the bispecific antibody binds to PD-1 and PD-L1. In some embodiments, the bispecific antibody that binds to PD-1 and PD-L1 is MCLA-136. In some embodiments, the bispecific antibody binds to PD-L1 and CTLA-4. In some embodiments, the bispecific antibody that binds to PD-L1 and CTLA-4 is AK104.

[0340] In some embodiments, the compounds of this disclosure may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitors are inhibitors of IDO1, TDO, or arginase. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099, and LY338196. An example of an arginase inhibitor is INCB1158.

[0341] As provided throughout, additional compounds, inhibitors, drugs, etc., may be combined with this compound in single or sequential dosage forms, or they may be administered simultaneously or sequentially in separate dosage forms.

[0342] The following are examples of how the present invention may be implemented. They should not be construed as limiting the scope of the present invention. [Examples]

[0343] Example 1. Characterization of cyclin E1 in ovarian cancer and endometrial cancer cell lines The cyclin E1 ("CCNE1") gene was evaluated in various ovarian and endometrial cancer cell lines (Figures 1A and 1B). CCNE1 was amplified in COV318, OVCAR3 ovarian, Fu-OV1, and KLE cells, each showing CCNE1 gain-of-function depending on the copy number (copy number ("CN") > 2) (Figure 1A). In contrast, CCNE1 was not amplified in COV504, OV56, or Igrov1 cells, each showing an intermediate copy number (2) or loss of function (CN ≤ 2). CN was obtained from the Broad Institute Cancer Cell Line Encyclopedia ("CCLE") database (Barretina, et al., Nature, 2012. 483(7391): p. 603-7, the entire database of which is incorporated herein by reference).

[0344] Western blot analysis was performed on protein samples from COV318, OVCAR3_ovary, Fu-OV1, KLE, COV504, OV56, and Igrov1 cells to assess CCNE1 protein levels. CCNE1 protein levels were higher in cell lines with CCNE1 gain-of-function due to copy number (CN>2, i.e., COV318, OVCAR3 ovary, Fu-OV1, and KLE cells) compared to cell lines with intermediate copy number or loss of function of the gene (CN≦2, i.e., COV504, OV56, and Igrov1 cells).

[0345] Example 2. CDK2 knockdown using siRNA inhibits the proliferation of CCNE1-amplified human cancer cell lines, but not of non-CCNE1-amplified human cancer cell lines. The effects of CDK2 knockdown were evaluated in CCNE1-amplified cell lines versus CCNE1-non-amplified cell lines. CCNE1-amplified cell lines (Fu-OV1 and KLE) or CCNE1-non-amplified cell lines (COV504 and Igrov1) were treated with a control ("ctrl") or CDK2-specific small interfering RNA ("siRNA") ("CDK2 siRNA-1" and "CDK2 siRNA-2") (Figures 2A and 2B, and 3A and 3B). 72 hours after siRNA transfection, cells were collected and subjected to cell cycle analysis by fluorescence-activated cell sorting ("FACS") (Figures 2A and 3A). CDK2 knockdown was confirmed by Western blotting (Figures 2B and 3B). CDK2 knockdown inhibited proliferation in CCNE1-amplified cell lines but not in CCNE1-non-amplified cell lines (Figures 2A and 3A).

[0346] Similar experiments were performed with additional CCNE1-amplified cell lines (COV318, OVCAR3, Fu-OV1, and KLE) and non-amplified CCNE1 cell lines (COV504, OV56, and Igrov1) (Figure 4). The percentage of S-phase cells 3 days after treatment with CDK2-specific siRNA was significantly reduced in CCNE1-amplified cell lines compared to treatment with control siRNA (Figure 4). Consistent with the results in Figures 2A and 3A, there was no significant difference in the percentage of S-phase cells 3 days after treatment with CDK2-specific siRNA compared to treatment with control siRNA in non-amplified CCNE1 cell lines (Figure 4).

[0347] Example 3. Proliferation in CCNE1-amplified and CCNE-non-amplified cell lines under CDK4 / 6 inhibition The effects of CDK4 / 6 inhibition were evaluated in CCNE1-amplified cell lines versus non-amplified CCNE1 cell lines. CCNE1-amplified cells (OVCAR3) or non-amplified CCNE1 cells (COV504) were treated with dimethyl sulfoxide ("DMSO") control or escalating concentrations of the CDK4 / 6 inhibitor palbociclib (Figure 5). Cells were harvested 16 hours after treatment with DMSO or palbociclib and subjected to FACS cell cycle analysis (Figure 5). CDK4 / 6 inhibition resulted in dose-dependent inhibition of proliferation in non-amplified CCNE1 cells, but not in CCNE1-amplified cells (Figure 5).

[0348] Similar experiments were performed with larger sets of CCNE1-amplified cell lines (COV318 and OVCAR3) and CCNE1-non-amplified cell lines (COV504, OV56, and Igrov1) (Figure 6). The percentage of S-phase cells 16 hours after palbociclib treatment decreased in a dose-dependent manner in CCNE1-non-amplified cell lines compared to DMSO treatment (Figure 6). Consistent with the results in Figure 5, there was no significant difference in the percentage of S-phase cells 16 hours after palbociclib treatment compared to DMSO treatment in CCNE1-amplified cell lines (Figure 6).

[0349] Example 4. CDK2 knockdown blocks Rb phosphorylation at S780 in CCNE1-amplified cell lines, but not in non-CCNE1-amplified cell lines. The effect of CDK2 knockdown on Rb phosphorylation at Ser-780 ("S780") of Sequence ID No. 3 in CCNE1-amplified cell lines versus CCNE1-non-amplified cell lines was evaluated. CCNE1-amplified cell lines (COV318, Fu-OV1, and KLE) or CCNE1-non-amplified cell lines (COV504, OV56, and Igrov1) were treated with ctrl or CDK2-specific siRNA (Figures 7A and 7B). 72 hours after siRNA transfection, cells were harvested, and total protein was extracted and analyzed by Western blotting. CDK2 knockdown was confirmed by Western blotting. CDK2 knockdown blocked Rb phosphorylation at S780 in CCNE1-amplified cell lines (Figure 7A), but not in CCNE1-non-amplified cell lines (Figure 7B).

[0350] Example 5. Palbociclib blocks Rb phosphorylation at S780 in non-CCNE1 amplified cell lines, but does not block it in CCNE1 amplified cell lines. The effect of CDK4 / 6 inhibition on Rb phosphorylation at S780 was evaluated in CCNE1-amplified cell lines versus non-amplified CCNE1 cell lines. CCNE1-amplified cell lines (OVCAR3 and COV318) or non-amplified CCNE1 cell lines (COV504 and OV56) were treated with DMSO or various doses of palbociclib (Figures 8A and 8B). Cells were harvested 1 or 15 hours after treatment, and total protein was extracted and analyzed by Western blotting (Figure 8). Palbociclib treatment blocked Rb phosphorylation at S780 in non-amplified CCNE1 cell lines (Figure 8B), but not in CCNE1-amplified cell lines (Figure 8A).

[0351] Example 6. CDK2 degradation by dTAG reduces RB phosphorylation at S780. To further confirm that CDK2 knockdown reduces Rb phosphorylation at S780 in CCNE1-amplified cells (see Example 4), CDK2 was degraded using a dTAG system and the level of S780-phosphorylated Rb was evaluated (Erb et al., Nature, 2017, 543(7644):270-274, the entire article is incorporated herein by reference). Briefly, OVCAR3 cells were engineered to express Cas9 by lentiviral transduction of a Cas9 construct. Subsequently, OVCAR3-Cas9 cells were engineered to express the CDK2-FKBP12F36V-HA fusion protein by lentiviral transduction of a CDK2-FKBP12F36V-HA expression construct. Next, in order to manipulate the cell line and inactivate endogenous CDK2, OVCAR3 (Cas9, CDK2-FKBP12F36V-HA) cells were transduced with CDK2 sgRNA ("CDK2-gRNA"), and OVCAR3 (Cas9, CDK2-FKBP12F36V-HA) cells transduced with untargeted sgRNA ("Ctl-gRNA", Cellecta) served as a control cell line.

[0352] Cells were treated with DMSO or dTAG concentration titration for 14 hours to degrade the CDK2-FKBP12F36V-HA protein with dTAG (Figure 9A). Cells were collected and processed for Western blotting (Figure 9B). Dose-responsive degradation of CDK2-FKBP12(F36V) was detected by Western blotting after dTAG treatment in both control and CDK2-gRNA treated cells (Figure 9B). Degradation was further confirmed by Western blotting of the HA tag. Endogenous CDK2 protein was detected in OVCAR3 cells treated with control gRNA but not in cells treated with CDK2-gRNA (Figure 9B). CDK2-FKBP12(F36V) degradation inhibited Rb phosphorylation at S780 in CDK2 knockout OVCAR3 cells but not in OVCAR3 cells with endogenous CDK2 expression.

[0353] Example 7. p-Rb S780 HTRF cell assay for identifying CDK2 inhibitors An in vitro CDK2 / CCNE1 enzyme activity assay was used to measure the phosphorylation of the peptide substrate using homogeneous time-resolved energy transfer ("HTRF"). First, the specificity of 8-((1R,2R)-2-hydroxy-2-methylcyclopentyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrido[2,3-d]pyrimidine-7(8H)-one (compound A) for CDK2 inhibition was confirmed by a kinase activity assay (Figure 10A). For this purpose, the LANCE® Ultra kinase assay was used with ULight®-labeled EIF4E-binding protein 1 (Thr37 / 46) peptide (PerkinElmer, TRF0128-M) as a substrate and with europium-labeled anti-phospho-EIF4E-binding protein 1 (Thr37 / 46) antibody (PerkinElmer, TRF0216-M). The ratio of the fluorescence transferred to the labeled substrate (665 nm) to the fluorescence of the europium donor (620 nm) represents the degree of phosphorylation. IC of compound A 50 This was determined to be 1.1 nM (Figure 10A). In contrast, the IC of the CDK4 / 6 inhibitor palbociclib 50 The value was 10,000 nM (Figure 10A).

[0354] Next, a CDK2 pRb(S780)HTRF cell assay was performed to enable quantitative detection of Rb phosphorylation at serine 780 in CCNE1-amplified COV318 cells upon treatment with compound A or palbociclib (Figure 10B). Treatment with compound A, rather than palbociclib, inhibited Rb phosphorylation at serine 780 in CCNE1-amplified cells (Figure 10B). IC of compound A in this assay 50 While the IC of palbociclib was 37nM, 50 The value was >3,000 nM (Figure 10B).

[0355] Example 8. Bioinformatics analysis of the CCLE dataset revealed that the sensitivity to CDK2 inhibition in CCNE1-amplified cells is dependent on functional p16. To identify biomarkers for predicting sensitivity to CDK2 inhibition in CCNE1-amplified cells, 460 cell lines from CCLE were analyzed (Barretina, above). First, cell lines were filtered based on CCNE1 copy number and expression based on shRNA knockdown data, as well as CDK2 sensitivity scores. In total, 41 cell lines were identified with a CCNE1 copy number and CCNE1 expression score >3 (CCLE: >3). Of these 41 cell lines, 18 (44%) were highly sensitive to CDK2 inhibition (CDK2 sensitivity score ≤ -3), while 23 (56%) were less sensitive (CDK2 sensitivity score > -3).

[0356] Next, the p16 status was evaluated in CDK2-sensitive and CDK2-insensitive cell lines (Figure 11). Of the 18 cell lines that showed high sensitivity to CDK2 inhibition, 100% expressed a normal p16 gene (Figure 11). In contrast, of the 23 CDK2-insensitive cell lines, only 4 expressed a normal p16 gene (Figure 11). The majority of the 23 CDK2-insensitive cell lines showed dysfunctional p16 gene expression: specifically, 10 of the 23 cell lines had a deletion of the p16 gene, 5 of the 23 cell lines had a silencing of the p16 gene, and 4 of the 23 cell lines had a mutation in the p16 gene (Figure 11).

[0357] Table 2 below provides an overview of CDK2 sensitivity and CDKN2A / p16 status in CCNE1-amplified cell lines. Table 2. Cell lines with a CDK2 sensitivity score of ≤3 were calculated as CDK2-sensitive cells, and those with a score of ≥3 were calculated as CDK2-insensitive cells. Cell lines validated in experiments are shown in bold. NCIN87_stomach showed no CDKN2A / P16 protein expression at all in Western blotting. The copy numbers of CCNE1 and CDKN2A / P16 were calculated based on the CCLE dataset. An expression score <0 was calculated as gene silencing. [Table 2]

[0358] Example 9. CCNE1-amplified cells with dysfunctional p16 do not respond to CDK2 inhibition. To further evaluate the role of CDK2-sensitive p16 in CCNE1-amplified cells, p16 protein expression was assessed by Western blotting in three gastric cell lines with CCNE1 amplification. AGS and NCI-N87 cells showed p16 levels deficient or dramatically reduced (Figure 12A). In contrast, p16 protein was detected in MKN1 cell protein extracts (Figure 12A).

[0359] Next, the effects of CDK2 knockdown in these cells were evaluated. Mkn1, Ags, and NCI-N87 cells were treated with either control or CDK2-specific siRNA. Three days after siRNA transfection, the cell cycle distribution of the cells was assessed by FACS. The proportion of S-phase cells in Mkn1 cells (CCNE1 amplified, p16 protein detected) was significantly reduced in CDK2 siRNA-treated cells compared to the control (Figure 12B). In contrast, the proportion of S-phase cells was not significantly reduced in Ags and NCI-N87 cells (CCNE1 amplified, dysfunctional p16 protein levels) after treatment with CDK2 siRNA compared to the control (Figure 12B).

[0360] Example 10. siRNA-mediated p16 knockdown neutralizes CDK2 inhibition-induced cell cycle repression in CCNE1-amplified cells. To confirm the role of p16 in CDK2 sensitivity of CCNE1-amplified cells, COV318 cells were treated with either a control or p16-specific siRNA. 72 hours after transfection, cells were treated with DMSO (control) or 100 nM compound A. 16 hours after treatment with DMSO or the CDK2 inhibitor, cells were harvested and subjected to FACS cell cycle analysis. Consistent with the results described above, the percentage of S-phase cells was significantly reduced in control siRNA-treated cells treated with the CDK2 inhibitor (compound A), but not in those treated with the DMSO control (Figure 13). In contrast, the percentage of S-phase cells was not significantly reduced in p16 knockdown cells after treatment with the CDK2 inhibitor (compound A) compared to the DMSO control (Figure 13).

[0361] Materials and methods used in Examples 1-10 Cell culture and transfection Human cyclin E1 (CCNE1)-amplified ovarian cell lines OVCAR3, COV318, Fu-OV1, endometrial cell line KLE, gastric cell lines MKN1, AGS, NCIN87, and CCNE1-non-amplified ovarian cell lines COV504, OV56, Igrov1 were cultured in RPMI 1640 medium. Complete growth medium was supplemented with 10% FBS, 0.1 mM non-essential amino acids, 2 mM L-glutamine, 100 units / mL penicillin G, and 100 μg / mL streptomycin in a humidified incubator at 37°C and in an atmosphere of 5% CO2 in the air. The Fu-OV1 cell line was purchased from the Leibniz-Institute DSMZ-German Collection of Microorganisms and Cell Cultures, MKN1 from the Japanese Cancer Research Resources Bank, and the remaining cell lines from the American Type Culture Collection. For transfection, cells were seeded in 6 wells for 24 hours and transiently transfected with Lipofectamine 2000 reagent (Thermo Fisher, 11668027). Endogenous CDK2 and CDKN2A / p16 were knocked down using ON-TARGETplus Human CKD2 siRNA (GE Healthcare Dharmacon, J-003236-11-0002 and J-003236-12-0002) and ON-TARGETplus Human CDKN2A / p16 siRNA (GE Healthcare Dharmacon, J-011007-08-0002). ON-TARGETplus Non-targeting Pool (GE Healthcare Dharmacon, D-001810-10-20) was used as a negative control.

[0362] Western blot analysis Whole cell extracts were prepared using RIPA buffer (Thermo Scientific, 89900) containing Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Scientific, 78440). Protein concentration was quantified using the BCA Protein Assay Kit (Thermo Scientific, 23225), and 40 μg of protein lysate was loaded onto a precast gradient gel (Bio-Rad, Hercules, No. 456-1094) for SDS-PAGE. The samples were diluted in 5x Laemmli buffer (300 mM Tris-HCl (pH 6.8), 10% SDS (w / v), 5% 2-mercaptoethanol, 25% glycerol (v / v), 0.1% bromophenol blue (w / v)) and boiled for 5 minutes. 35 μg of protein was separated by 8-15% SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane. Nonspecific binding sites on the PVDF membrane were blocked with 5% skim milk in TBST (20 mM Tris-HCl (pH 7.6), 137 mM NaCl, 1% Tween-20). The membranes were hybridized overnight at 4°C with antibodies against anti-CDKN2A / p16 (Cell Signaling Technology, 92803S), anti-Cas9 (Cell Signaling Technology, 97982S), anti-HA (Cell Signaling Technology, 3724S), anti-Rb (Cell Signaling Technology, 9309S), anti-phospho-Rb (Ser780) (Cell Signaling Technology, 8180S), anti-CDK2 (Cell Signaling Technology, 2546S), anti-CCNE1 (Cell Signaling Technology, 20808S), and anti-GAPDH (Cell Signaling Technology, 8884S), followed by incubation at room temperature for 1 hour with horseradish peroxidase (HRP)-complexed secondary antibodies. The membranes were then developed using Immobilon Western chemiluminescent HRP substrate (Millipore, WBKLS0500).Images were captured using the Odyssey CLx Imager (LI-COR) emission / fluorescence imaging system.

[0363] Cell cycle analysis Cells were seeded in 6-well tissue culture plates and treated 24 hours later with concentration titration of palbociclib or compound A. After overnight treatment, cells were exposed to 10 μM EdU for 3 hours, and EdU-DNA was detected using the Click-iT AlexaFluor® 647 azide kit (Life Technology, C10424) according to the manufacturer's instructions. Bulk DNA was stained with DAPI. Compound-treated and DMSO-treated control cells were acquired using CytoFlex (Beckman Coulter) and analyzed using FlowJo software. For cell cycle analysis of cells with siRNA knockdown, cells were exposed to 10 μM EdU for 3 hours 72 hours after siRNA transfection, and then detected using the Click-iT Alexa Fluor® 647 azide kit.

[0364] plasmid The LentiCas9 plasmid pRCCH-CMV-Cas9-2A (Cellecta, SVC9-PS) was used for Cas9 expression. An sgRNA-CDK2 lentiviral construct designed to target the CDK2 AAGCAGAGATCTCTCGGA (SEQ ID NO: 8) was cloned into the sgRNA expression vector pRSG-U6, which was purchased from Cellecta (93661). For CDK2-FKBP12F36V-HA expression, a 1306 base pair DNA fragment encoding CDK2 and the C-terminal FKBP12F36V-2xHA tag was synthesized and cloned into the EcoRI and BamHI digested pCDH-EF1α-MCS-T2A-Puro lenti vector (Systembio, CD527A-1).

[0365] Sequence of a 1306bp DNA fragment: [ka]

[0366] The restriction enzyme sites GAATTC (SEQ ID NO: 5, EcoRI), GGATCC (SEQ ID NO: 6, BamHI), and TTCGAA (SEQ ID NO: 7, BstBI) are underlined. Sequences encoding CDK2 are in bold, and the FKBP12F36V-HA sequence is italicized. The three underlined nucleic acids within the CDK2 sequence show modifications that omit the PAM site to avoid the CRISPR knockout effect. These modifications did not alter the encoded amino acids.

[0367] lentivirus production Lentiviruses were generated in 293T cells by co-transfection with Lentiviral Packaging Mix (Sigma, SHP001) and a given lentiviral expression plasmid using Lipofectamine 2000. The viral supernatant was collected 48 and 72 hours after transfection and filtered through a 0.22 μm membrane. All cell lines were transduced by spinoculation at 2000 rpm for 1 hour at room temperature using 8 μg / mL of polyblen (Santa Cruz, sc-134220).

[0368] CDK2-dTAG cells OVCAR3 cells were initially engineered to express Cas9 by lentiviral transduction of a Cas9 construct. Selected cells were maintained in 100 μg / mL hygromycin (Life Technologies, 10687010) and Cas9 expression was verified by immunoblotting. Subsequently, OVCAR3-Cas9 cells were engineered to express the CDK2-FKBP12F36V-HA fusion protein by lentiviral transduction of a CDK2-FKBP12F36V-HA expression construct and selection with 2 μg / mL puromycin dihydrochloride (Life Technologies, A1113803). CDK2-FKBP12F36V-HA expression was verified by immunoblotting using anti-CDK2 and anti-HA antibodies. Next, to manipulate the cell line and inactivate endogenous CDK2, OVCAR3 (Cas9, CDK2-FKBP12F36V-HA) cells were transduced with CDK2 sgRNA and selected with 50 μg / mL Zeocin (Life Technologies, R25001). Inactivation of endogenous CDK2 expression in the proliferated clones was tested by immunoblotting. OVCAR3 (Cas9, CDK2-FKBP12F36V-HA) cells transduced with non-target sgRNA (Cellecta) served as a control cell line.

[0369] To degrade the CDK2-FKBP12F36V-HA protein by dTAG, 200,000 cells were seeded in 1 mL of medium in triples in 24-well plates and treated with dimethyl sulfoxide (DMSO) or dTAG concentration titration for 14 hours. Cells were collected and processed for Western blotting.

[0370] CDK2 / CCNE1 Enzyme Assay The in vitro CDK2 / CCNE1 enzyme activity assay measures the phosphorylation of peptide substrates using homogeneous time-resolved energy transfer (HTRF). The LANCE® Ultra kinase assay used ULight®-labeled EIF4E-binding protein 1 (Thr37 / 46) peptide (PerkinElmer, TRF0128-M) as a substrate and europium-labeled anti-phospho-EIF4E-binding protein 1 (Thr37 / 46) antibody (PerkinElmer, TRF0216-M). The ratio of the fluorescence transferred to the labeled substrate (665 nm) to the fluorescence of the europium donor (620 nm) represents the degree of phosphorylation. The ratio of treated wells is normalized to DMSO only (100% activity) and no enzyme (0% activity) controls. Normalized data are analyzed using dose-response curves for four parameters, and the IC of each compound is calculated. 50 To decide.

[0371] CDK2 pRb(S780) HTRF Cell Assay The CDK2 pRb(S780)HTRF cell assay enables quantitative detection of Rb phosphorylated on serine 780 in CCNE1-amplified COV318 cells. This assay included two antibodies: europium cryptotate-labeled anti-phospho-Rb S780 antibody (donor) and d2-labeled anti-Rb antibody (acceptor). Briefly, COV318 cells were seeded in 96-well plates at a density of 25,000 cells per well with nine steps of 3-fold serially diluted compounds and cultured overnight at 37°C in 5% CO2. The final compound concentration was started at 3 μM. The following day, cells were lysed in 70 μL of 1x Phospho-total protein lysis buffer #2 (Cisbio) supplemented with 0.7 μL of blocking buffer (Cisbio) and 1.4 μL of protease inhibitor cocktail set III, EDTA-free (Calbiochem, 539134). 16 μL of the cell lysate was mixed with 4 μL of fluorophore-conjugated antibody to final concentrations of 0.188 nM cryptotate-labeled anti-phospho-Rb S780 antibody and 0.14 nM d2-labeled anti-Rb antibody. After incubation at room temperature for 2 hours, the HTRF signal was measured on a PHERAstar microplate reader (BMG Labtech) using a 340 nm excitation wavelength, a 620 nm filter for europium donor fluorescence, and a 665 nm filter for acceptor fluorescence detection. The HTRF signal was calculated as the HTRF ratio (ratio of fluorescence measured at 665 nm and 620 nm) × 10000.

[0372] Example A1.4-((8-cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d[pyrimidine-2-yl)amino)benzenesulfonamide [ka] Step 1: 5-Bromo-N-cyclopentyl-2-methoxypyrimidine-4-amine [ka] To a solution of 5-bromo-2,4-dichloropyrimidine (3.08 mL, 24.05 mmol) in THF (80 mL), cyclopentanamine (2.62 mL, 26.5 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours and then filtered. The filtrate was concentrated and dissolved in sodium methoxide in MeOH (21% w / w, 3 mL), and then heated under reflux for 2 hours. The mixture was diluted with water and ethyl acetate, and the layers were separated. The organic layer was washed with water and brine, dried over sodium sulfate, and concentrated. The residue was purified with Biotage Isolera® (0-50% ethyl acetate in hexane) to obtain the desired product as a white solid (4.7 g, 72%). 10 H 15 BrN3O (M+H) + LCMS calculated value for this: m / z = 272.0 / 274.0; measured value: 272.0 / 274.0.

[0373] Step 2.3-(4-(cyclopentylamino)-2-methoxypyrimidine-5-yl)propanoate ethyl [ka] A mixture of 5-bromo-N-cyclopentyl-2-methoxypyrimidine-4-amine (500 mg, 1.837 mmol), triethylamine (512 μL, 3.67 mmol), ethyl acrylate (300 μL, 2.76 mmol), and tetrakis(triphenylphosphine)palladium (0) (212 mg, 0.184 mmol) was mixed with DMF (6 mL), the reaction flask was evacuated, nitrogen was returned, and the flask was packed, then stirred overnight at 120 °C. The mixture was then poured into ethyl acetate / water, and the layers were separated. The aqueous layer was extracted with ethyl acetate, the combined organic matter was washed with water and brine, dried over sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera (trademark) (0-100% ethyl acetate in hexane). The intermediate was dissolved in EtOH (6 mL), and palladium carbon (10%, 391 mg, 0.367 mmol) was added. The reaction flask was evacuated and then filled with hydrogen gas through the balloon. The reaction mixture was stirred at room temperature for 3 hours, then diluted with ethyl acetate and filtered through a Celite plug. The filtrate was concentrated and used in the next step without further purification of the crude product (340 mg, 63%). 15 H 24 N3O3(M+H) + LCMS calculated value for this: m / z = 294.2; measured value: 294.2.

[0374] Step 3. 8-Cyclopentyl-2-methoxy-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one [ka] To a solution of ethyl 3-(4-(cyclopentylamino)-2-methoxypyrimidine-5-yl)propanoate (5.0 g, 17.04 mmol) in THF (28 mL) / water (28 mL), lithium hydroxide hydrate (1.073 g, 25.6 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes, then quenched with HCl (12 N, 2.13 mL, 25.6 mmol) and concentrated. The crude product was dissolved in DMF (4 mL) and HATU (7.13 g, 18.75 mmol), and Hünig base (5.95 mL, 34.1 mmol) was added. The reaction mixture was then stirred at room temperature for 2 hours, quenched with water, and extracted with ethyl acetate. The organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera (trademark) (20-100% ethyl acetate in hexane) to obtain the desired product (2.01 g, 48%). 13 H 18 N3O2(M+H) + LCMS calculated value for this: m / z = 248.2; measured value: 248.2.

[0375] Step 4. 8-Cyclopentyl-2-methoxy-6,6-dimethyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one [ka] To a solution of 8-cyclopentyl-2-methoxy-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one (501 mg, 2.026 mmol) in DMF (10 mL), methyl iodide (380 μL, 6.08 mmol) and sodium hydride (60% in mineral oil, 284 mg, 7.09 mmol) were added, and the reaction mixture was heated to 65°C for 2 hours. The mixture was quenched with water and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over sodium sulfate, and concentrated. The crude residue was purified with Biotage Isolera® (0-100% ethyl acetate in hexane) to obtain the desired product as a colorless oil (303 mg, 54%). 15 H 22 N3O2(M+H) +LCMS calculated value for this: m / z = 276.2; measured value: 276.2.

[0376] Step 5. 8-Cyclopentyl-6,6-dimethyl-7-oxo-2,3,5,6,7,8-hexahydropyrido[2,3-d]pyrimidine-2-yltrifluoromethanesulfonate [ka] To a solution of 8-cyclopentyl-2-methoxy-6,6-dimethyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one (131 mg, 0.476 mmol) in acetonitrile (2.4 mL), sodium iodide (143 mg, 0.952 mmol) and TMS-Cl (122 μL, 0.952 mmol) were added. The reaction mixture was stirred overnight at room temperature, then quenched with water and extracted with ethyl acetate. The organic layer was washed with saturated sodium thiosulfate aqueous solution, water, and brine, dried on sodium sulfate, and concentrated. The crude product was dissolved in DCM (2.5 mL) and pyridine (42.3 μL, 0.523 mmol) was added. The reaction mixture was cooled to 0°C, and trifluoromethanesulfonic acid anhydride (96 μL, 0.571 mmol) was added dropwise. Next, the reaction mixture was heated to room temperature and stirred for 2 hours, then quenched with saturated sodium bicarbonate and extracted with DCM. The organic layer was dried over sodium sulfate and concentrated. The crude product was used in the next step without further purification (141 mg, 75%). 15 H 21 F3N3O4S (M+H) + LCMS calculated value for this: m / z = 396.2; measured value: 396.2.

[0377] Step 6.4-((8-cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine-2-yl)amino)benzenesulfonamide 8-Cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine-2-yltrifluoromethanesulfonate (20 mg, 0.051 mmol), 4-aminobenzenesulfonamide (17.51 ​​mg, 0.102 mmol), XantPhos Pd G2 (4.52 mg, 5.08 μmol), and potassium carbonate (70.3 mg, 0.508 mmol) were mixed with 1,4-dioxane (508 μL). The reaction flask was evacuated, nitrogen was returned, and the mixture was packed, then stirred at 100°C for 2 hours. The mixture was then diluted with MeOH and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min). 20 H 26 N5O3S (M+H) + LCMS calculated value for this: m / z = 416.2; measured value: 416.2.

[0378] Example A2.8-Cyclopentyl-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d[pyrimidine-7(6H)-one [ka] This compound was prepared in the same manner as in step 6 of Example A1, using 1-(methylsulfonyl)piperidine-4-amine instead of 4-aminobenzenesulfonamide and RuPhos Pd G2 instead of XantPhos Pd G2. 20 H 32 N5O3S (M+H) + LCMS calculated value for this: m / z = 422.2; measured value: 422.2. 1H NMR (600 MHz, DMSO) δ 8.01 (s, 1H), 5.44 - 5.22 (m, 1H), 3.85 (bs, 1H), 3.59 (d, J = 12.3 Hz, 1H), 2.9 (s, 3H), 2.85 (t, J = 12.2, 2.6 Hz, 1H), 2.60 (s, 2H), 2.05 (s, 1H), 1.98 (d, J = 16.3 Hz, 1H), 1.93 - 1.87 (m, 1H), 1.74 (s, 1H), 1.59 (m, 2H), 1.09 (s, 6H).

[0379] Example A3. 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-phenyl-5,8-dihydropyrido[2,3-d[pyrimidine-7(6H)-one [ka] Step 1. Dimethyl 2,2-dimethylpentanedioate [ka] To a solution of 3,3-dimethyldihydro-2H-pyran-2,6(3H)-dione (10 g, 70.3 mmol) in methanol (100 mL), 10 drops of concentrated sulfuric acid were added, and the reaction mixture was heated overnight to 60°C. The mixture was then concentrated. The residue was diluted with ethyl acetate, washed with saturated sodium bicarbonate and brine, and then dried over sodium sulfate and concentrated. The crude product was used in the next step without further purification.

[0380] Step 2.3-(2-amino-6-oxo-1,6-dihydropyrimidine-5-yl)-2,2-methyl dimethylpropanoate [ka] To a solution of diisopropylamine (5.32 mL, 37.4 mmol) in THF (12 mL) at -78°C, n-BuLi (2.5 M in hexane, 14.94 mL, 37.4 mmol) was added dropwise, and the reaction mixture was stirred at -78°C for 1 hour. Next, a solution of dimethyl 2,2-dimethylpentanedioate (5.86 g, 31.1 mmol) in THF (20 mL) was added dropwise, and the reaction mixture was stirred at -78°C for a further 1.5 hours. Then, methyl formate (2.88 mL, 46.7 mmol) was added, and the reaction mixture was stirred at -78°C for 1 hour, followed by quenching with saturated ammonium chloride. After warming to room temperature, the mixture was diluted with ethyl acetate / water, and the layers were separated. The organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The residue was dissolved in MeOH (10 mL), and guanidine carbonate (5.61 g, 31.1 mmol) was added. The reaction mixture was heated overnight to 60°C, then concentrated and purified with Biotage Isolera® (2-12% methanol in dichloromethane) to obtain the desired product as a white solid (2.45 g, 35%). 10 H 16 N3O3(M+H) + LCMS calculated value for this: m / z = 226.2; measured value: 226.2.

[0381] Step 3. 3-(4-chloro-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrimidine-5-yl)-2,2-methyldimethylpropanoate [ka] 3-(2-amino-6-oxo-1,6-dihydropyrimidine-5-yl)-2,2-methylpropanoate (2.45 g, 10.88 mmol) was dissolved in POCl3 (10 mL), heated overnight to 100°C, and then slowly added to saturated sodium bicarbonate. The mixture was extracted with DCM, the organic layer was washed with saturated sodium bicarbonate and brine, and dried over sodium sulfate to concentrate. DMF (36.3 mL), 1-(methylsulfonyl)piperidine-4-one (2.506 g, 14.14 mmol), TFA (5.03 mL, 65.3 mmol), and sodium triacetoxyborohydride (5.76 g, 27.2 mmol) were added to this intermediate, and the reaction mixture was stirred at room temperature for 5 hours, then quenched with saturated sodium bicarbonate and extracted with DCM. The organic layer was washed with water and brine, and dried over sodium sulfate to concentrate. The residue was purified with Biotage Isolera (trademark) (2-12% methanol in DCM), and the desired product was obtained as a yellow solid (2.2 g, 50%). 16 H 26 ClN4O4S (M+H) + LCMS calculated value for this: m / z = 404.2 / 406.2; measured value: 404.2 / 406.2.

[0382] Step 4. 6,6-Dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-phenyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one 3-(4-chloro-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrimidine-5-yl)-2,2-methylpropanoate methyl (21 mg, 0.052 mmol), aniline (9.47 μL, 0.104 mmol), Ruphos Pd G2 (4.03 mg, 5.19 μmol), and cesium carbonate (50.7 mg, 0.156 mmol) were mixed with 1,4-dioxane (519 μL). The reaction flask was evacuated, nitrogen was returned, and the flask was packed, then stirred overnight at 100°C. The reaction mixture was diluted with MeOH and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min). 21 H 28 N5O3S (M+H) + LCMS calculated value for this: m / z = 430.2; measured value: 430.2.

[0383] Example A4. 8-(1,1-difluorobutan-2-yl)-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one [ka] This compound was prepared using 1,1-difluorobutan-2-amine as a coupling partner in the same manner as in step 4 of Example A3. The product was isolated as a racemic mixture. 19 H 30 F2N5O3S (M+H) + LCMS calculated value for this: m / z = 446.2; measured value: 446.2.

[0384] Example A5. 6,6-dimethyl-8-((1-methyl-1H-pyrazole-5-yl)methyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one [ka] This compound was prepared using (1-methyl-1H-pyrazole-5-yl)methaneamine as a coupling partner in the same manner as in step 4 of Example A3. 20 H 30 N7O3S (M+H) + LCMS calculated value for this: m / z = 448.2; measured value: 448.2.

[0385] Example A6. 6,6-Dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-(tetrahydrofuran-3-yl)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one [ka] This compound was prepared using tetrahydrofuran-3-amine as a coupling partner in the same manner as in step 4 of Example A3. The product was obtained in racemic form. 19 H 30 N5O4S (M+H) + LCMS calculated value for this: m / z = 424.2; measured value: 424.2.

[0386] Example B1. 7'-Cyclopentyl-2'-((2-methyl-1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] Step 1: 5-Bromo-2-chloro-N-cyclopentylpyrimidine-4-amine [ka] To a solution of 5-bromo-2,4-dichloropyrimidine (20 g, 88 mmol) and Hünig base (22.99 mL, 132 mmol) in THF (219 mL), cyclopentanamine (9.56 mL, 97 mmol) was added. The reaction mixture was stirred overnight at room temperature, then quenched with water and extracted with ethyl acetate. The organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The residue was purified with Biotage Isolera® (0-40% ethyl acetate in hexane) to obtain the desired product as a yellow solid (21.1 g, 87%). C9H 12 BrClN3(M+H) + LCMS calculated value for this: m / z = 276.0 / 278.0; measured value: 276.0 / 278.0.

[0387] Step 2. (2-(tert-butoxy)-2-oxoethyl)zinc bromide(II) [ka] The zinc powder was washed in 2% HCl for 1 hour, and then the zinc was activated by decanting. Water was added to the solid, and the supernatant was decanted three times. The solid was then collected by filtration, washed with water, ethanol, acetone, and ether, and then dried in an oven for 15 minutes. To a portion of the zinc thus prepared (4.87 g, 74.4 mmol), THF (65 mL) and TMS-Cl (0.865 mL, 6.77 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour, and then tert-butyl 2-bromoacetate (10.00 mL, 67.7 mmol) was added dropwise. The addition was completed over approximately 15 minutes. The mixture was then heated to 50°C for 1 hour, at which point most of the zinc metal had dissolved. The mixture was cooled to room temperature and used as a solution of approximately 0.9 M in the subsequent steps.

[0388] Step 3: 2-(2-chloro-4-(cyclopentylamino)pyrimidine-5-yl) tert-butyl acetate [ka] To a mixture of 5-bromo-2-chloro-N-cyclopentylpyrimidine-4-amine (10 g, 36.2 mmol), Pd2(dba)3 (0.993 g, 1.085 mmol), and 1,2,3,4,5-pentaphenyl-1'-(di-t-butylphosphino)ferrocene (QPhos, 0.771 g, 1.085 mmol), (2-(tert-butoxy)-2-oxoethyl)zinc bromide (II) (48.2 mL, 43.4 mmol) and dioxane (72 mL) were added as a freshly prepared 0.9 M solution in THF. The mixture was vacuumed, nitrogen was returned, and the mixture was packed, then stirred at room temperature for 1 hour. The reaction product was quenched with 1N HCl and extracted with ethyl acetate. The organic layer was washed with water and brine and concentrated. The crude product was purified with Biotage Isolera (trademark) (0-50% ethyl acetate in hexane) to obtain the desired product as a pink solid (7.6 g, 67%). 15 H 23 ClN3O2(M+H) + LCMS calculated value for this: m / z = 312.2; measured value: 312.2.

[0389] Step 4. 2-Chloro-7-cyclopentyl-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidine-6-one [ka] To a solution of tert-butyl 2-(2-chloro-4-(cyclopentylamino)pyrimidine-5-yl)acetate (2.41 g, 7.73 mmol) in THF (25.8 mL), sodium hydride (60% in mineral oil, 0.618 g, 15.46 mmol) was added. The reaction mixture was heated to 60°C for 1 hour, then cooled to room temperature and quenched with 1N HCl. The mixture was extracted with ethyl acetate, the organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera® (0-100% ethyl acetate in hexane) to obtain the desired product as a green solid (1.46 g, 79%). 11 H 13 ClN3O (M+H) +LCMS calculated value for this: m / z = 238.2; measured value: 238.2.

[0390] Step 5. 2'-Chloro-7'-cyclopentylspiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one [ka] To a suspension of sodium hydride (60% in mineral oil, 0.981 g, 24.54 mmol) in THF (20 mL) / HMPA (2 mL, 11.50 mmol), a solution of 2-chloro-7-cyclopentyl-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidine-6-one (1.458 g, 6.13 mmol) in THF (2.5 mL) was added dropwise, and the reaction mixture was stirred at room temperature for 10 minutes. 1,2-dibromoethane (1.057 mL, 12.27 mmol) was added, and the reaction mixture was heated to 50°C for 1 hour, then quenched with 1N HCl and extracted with ethyl acetate. The organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera (trademark) (15-100% ethyl acetate in hexane) to obtain the desired product as an off-green solid (1.1 g, 68%). C 13 H 15 ClN3O (M+H) + LCMS calculated value for this: m / z = 264.2; measured value: 264.2.

[0391] Step 6.4-((7'-cyclopentyl-6'-oxo-6',7'-dihydrospiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-2'-yl)amino)-2-methylpiperidine-1-carboxylate tert-butyl [ka] Vials containing 2'-chloro-7'-cyclopentylspiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one (0.04 g, 0.152 mmol), 4-amino-2-methylpiperidine-1-carboxylate tert-butyl (0.098 g, 0.455 mmol), RuPhos Pd G2 (0.012 g, 0.015 mmol), and cesium carbonate (0.148 g, 0.455 mmol) were vacuumed and filled with nitrogen. 1,4-dioxane (1.996 mL) was added, and the solution was stirred at 100°C for 48 hours. The mixture was cooled and concentrated under reduced pressure, and purified with Teledyne ISCO CombiFlash® Rf+ (0-100% ethyl acetate in hexane) to obtain the desired product as red oil. 24 H 36 N5O3(M+H) + LCMS calculated value for this: m / z = 442.3; measured value: 442.3.

[0392] Step 7. 7'-Cyclopentyl-2'-((2-methylpiperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one hydrochloride [ka] A solution of 4-((7'-cyclopentyl-6'-oxo-6',7'-dihydrospiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-2'-yl)amino)-2-methylpiperidine-1-carboxylate tert-butyl (0.0163 g, 0.037 mmol) and 4M HCl (0.157 mL, 0.628 mmol) in dioxane was stirred at room temperature (rt) for 1 hour. The solution was then concentrated under reduced pressure. Toluene was added and the solution was concentrated under reduced pressure to obtain the desired product as an orange oil. 19 H 28 N5O (M+H) + LCMS calculated value for this: m / z = 342.2; measured value: 342.2.

[0393] Step 8. 7'-Cyclopentyl-2'-((2-methyl-1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one Methanesulfonyl chloride (6.06 μL, 0.078 mmol) was added dropwise at 0°C to a solution of 7'-cyclopentyl-2'-((2-methylpiperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one (0.022 g, 0.065 mmol) and Et3N (10.84 μL, 0.078 mmol) in anhydrous CH2Cl2 (1.866 mL). The solution was gradually warmed to room temperature overnight. The solution was then diluted with MeOH and CH3CN and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min) to obtain the desired product as a white solid. 20 H 30 N5O3S (M+H) + LCMS calculated value for this: m / z = 420.2; measured value: 420.5.

[0394] Example B2. 7'-Cyclopentyl-2'-((1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using 1-(methylsulfonyl)piperidine-4-amine as the amine coupling partner, in the same manner as in step 5 of Example B1. 19 H 28 N5O3S (M+H) + LCMS calculated value for this: m / z = 406.2; measured value: 406.2.

[0395] Example B3. 7'-Cyclopentyl-2'-((1-(cyclopropylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] Step 1.4-((7'-cyclopentyl-6'-oxo-6',7'-dihydrospiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-2'-yl)amino)piperidine-1-carboxylate tert-butyl [ka] This compound was prepared using 4-aminopiperidine-1-carboxylate tert-butyl as an amine coupling partner, in the same manner as in step 5 of Example B1. 23 H 34 N5O3(M+H) + LCMS calculated value for this: m / z = 428.3; measured value: 428.3.

[0396] Step 2: 7'-Cyclopentyl-2'-(Piperidine-4-ylamino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one [ka] A solution of 4-((7'-cyclopentyl-6'-oxo-6',7'-dihydrospiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-2'-yl)amino)piperidine-1-carboxylate tert-butyl (0.0599 g, 0.140 mmol) in a 1:1 TFA (0.05 mL) / CH2Cl2 (0.050 mL) mixture was stirred at room temperature for 1 hour. The reaction mixture was then quenched with saturated NaHCO3 and extracted in CH2Cl2 (2x). The organic layer was washed with brine, and the solution was concentrated under reduced pressure to obtain the desired product as a brown solid, which was used without further purification. 18 H 26 N5O (M+H) +LCMS calculated value for this: m / z = 328.2; measured value: 328.4.

[0397] Step 3: 7'-Cyclopentyl-2'-((1-(cyclopropylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one A solution of 7'-cyclopentyl-2'-(piperidine-4-ylamino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one (0.0115 g, 0.035 mmol), cyclopropanesulfonyl chloride (7.16 μL, 0.070 mmol), and Hünig base (0.015 mL, 0.088 mmol) in anhydrous THF (0.702 mL) was stirred overnight at room temperature. The solution was then diluted with CH3CN and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min) to obtain the desired product as a white solid. 21 H 30 N5O3S (M+H) + LCMS calculated value for this: m / z = 432.2; measured value: 432.2.

[0398] Example B4.7'-Cyclopentyl-2'-((1-((tetrahydro-2H-pyran-4-yl)sulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using tetrahydro-2H-pyran-4-sulfonyl chloride as the sulfonyl chloride, in the same manner as in step 3 of Example B3. 23 H 34 N5O4S (M+H) + LCMS calculated value for this: m / z = 476.2; measured value: 476.2.

[0399] Example B5. 7'-Cyclopentyl-2'-((1-(pyridine-3-ylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using pyridine-3-sulfonyl chloride hydrochloride as the sulfonyl chloride, in the same manner as in step 3 of Example B3. 23 H 29 N6O3S (M+H) + LCMS calculated value for this: m / z = 469.2; measured value: 469.2.

[0400] Example B6. 2'-((1-((4-chlorophenyl)sulfonyl)piperidine-4-yl)amino)-7'-cyclopentylspiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using 4-chlorobenzenesulfonyl chloride as the sulfonyl chloride, in the same manner as in step 3 of Example B3. 24 H 29 ClN5O3S (M+H) + LCMS calculated value for this: m / z = 502.2; measured value: 502.2.

[0401] Example B7.7'-Cyclopentyl-2'-((1-((1-methyl-1H-pyrazole-4-yl)sulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using 1-methyl-1H-pyrazole-4-sulfonyl chloride as the sulfonyl chloride, in the same manner as in step 3 of Example B3. 22 H30 N7O3S (M+H) + LCMS calculated value for this: m / z = 472.2; measured value: 472.4.

[0402] Example B8. 7'-(2-methylcyclopentyl)-2'-((1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] Step 1. 1-(4-chloro-2-(methylthio)pyrimidine-5-yl)cyclopropane-1-carboxylate ethyl [ka] To a suspension of sodium hydride (2.006 g, 50.2 mmol) in DMF (60 mL) at 0°C, a solution of 1,2-dibromoethane (2.59 mL, 30.1 mmol) and 2-(4-chloro-2-(methylthio)pyrimidine-5-yl)ethyl acetate (4.95 g, 20.06 mmol) in DMF (40 mL) was added dropwise. The reaction mixture was heated to room temperature and stirred for 30 minutes, then quenched with saturated ammonium chloride and extracted with ethyl acetate. The organic layer was washed with water and brine, dried on sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera® (0-50% ethyl acetate in hexane) to obtain the desired product as yellow oil (3.2 g, 59%). 11 H 14 ClN2O2S (M+H) + LCMS calculated value for this: m / z = 273.1; measured value: 273.1.

[0403] Step 2.1-(4-chloro-2-(methylsulfonyl)pyrimidine-5-yl)cyclopropane-1-carboxylate ethyl [ka] To a solution of ethyl 1-(4-chloro-2-(methylthio)pyrimidine-5-yl)cyclopropane-1-carboxylate (3.1 g, 11.37 mmol) in DCM (60 mL), m-CPBA (5.88 g, 34.1 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The mixture was then quenched with saturated sodium bicarbonate and extracted with DCM. The organic layer was washed with saturated sodium bicarbonate and brine, dried over sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera® (0-100% ethyl acetate in hexane) to obtain the desired product as a white solid. 11 H 14 ClN2O4S (M+H) + LCMS calculated value for this: m / z = 305.1; measured value: 305.1.

[0404] Step 3. 1-(4-chloro-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrimidine-5-yl)cyclopropane-1-carboxylate ethyl [ka] To a suspension of 1-(methylsulfonyl)piperidine-4-amine (2.226 g, 12.49 mmol) in tetrahydrofuran (56.8 mL) at 0°C, isopropylmagnesium chloride lithium chloride complex (10.48 mL, 13.62 mmol) was added, and the reaction mixture was stirred at 0°C for 30 minutes. Next, a solution of 1-(4-chloro-2-(methylsulfonyl)pyrimidine-5-yl)cyclopropane-1-carboxylate ethyl (3.46 g, 11.35 mmol) in THF was added dropwise, and the reaction mixture was heated to room temperature and stirred for 10 minutes, then heated to 55°C for 1 hour. The reaction mixture was quenched with saturated ammonium chloride and extracted with ethyl acetate. The organic layer was washed with water and brine, dried over sodium sulfate, and concentrated. The crude product was purified with Biotage Isolera® (15-100% ethyl acetate in hexane) to obtain the desired product as a white solid. 16 H 24 ClN4O4S (M+H) +LCMS calculated value for this: m / z = 403.2; measured value: 403.2

[0405] Step 4. 7'-(2-methylcyclopentyl)-2'-((1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine]-6'(7'H)-one 1,4-Dioxane (496 μL) was added to a mixture of ethyl 1-(4-chloro-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrimidine-5-yl)cyclopropane-1-carboxylate (20 mg, 0.050 mmol), 2-methylcyclopenylamine (10 mg, 0.99 mmol), RuPhos Pd G2 (3.86 mg, 4.96 μmol), and cesium carbonate (48.5 mg, 0.149 mmol). The reaction flask was evacuated, nitrogen was returned, and the flask was packed, then stirred at 140°C for 1.5 hours. The mixture was diluted with MeOH and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min) to obtain the desired product as a mixture of four diastereomers. 20 H 30 N5O3S (M+H) + LCMS calculated value for this: m / z = 420.2; measured value: 420.2.

[0406] Example B9. 2'-((1-(methylsulfonyl)piperidine-4-yl)amino)-7'-(o-tolyl)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] 1,4-Dioxane (165 μL) was added to a mixture of 1-(4-chloro-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrimidine-5-yl)cyclopropane-1-carboxylate ethyl (Example B6, Step 2, 20 mg, 0.050 mmol), XantPhos Pd G2 (4.41 mg, 4.96 μmol), o-toluidine (10.53 μL, 0.099 mmol), and cesium carbonate (81 mg, 0.248 mmol). The reaction flask was evacuated, nitrogen was returned, and the flask was packed, then stirred overnight at 120°C. The mixture was diluted with MeOH and purified by prep-LCMS (XBridge C18 column, eluted with an acetonitrile / water gradient containing 0.1% TFA at a flow rate of 60 mL / min). 21 H 26 N5O3S (M+H) + LCMS calculated value for this: m / z = 428.2; measured value: 428.2.

[0407] Example B10.7'-(1,1-difluorobutan-2-yl)-2'-((1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d[pyrimidine[-6'(7'H)-one [ka] This compound was prepared using 1,1-difluorobutane-2-amine hydrochloride as the amine coupling partner, in the same manner as in Example B8. The product was isolated in racemic form. 18 H 26 F2N5O3S (M+H) + LCMS calculated value for this: m / z = 430.2; measured value: 430.2.

[0408] Example B11. 7'-(1,5-dimethyl-1H-pyrazole-4-yl)-2'-((1-(methylsulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine[-6'(7'H)-one [ka] This compound was prepared using 1,5-dimethyl-1H-pyrazole-4-amine as a coupling partner, in the same manner as in Example B9. 19 H 26 N7O3S (M+H) + LCMS calculated value for this: m / z = 432.2; measured value: 432.2.

[0409] Example B12.7'-((1R,3R)-3-hydroxycyclohexyl)-2'-((1-((1-methyl-1H-pyrazole-4-yl)sulfonyl)piperidine-4-yl)amino)spiro[cyclopropane-1,5'-pyrrolo[2,3-d]pyrimidine[-6'(7'H)-one [ka] Step 1.4-((4-chloro-5-(1-(ethoxycarbonyl)cyclopropyl)pyrimidine-2-yl)amino)piperidine-1-carboxylate benzyl [ka] To a solution of ethyl 1-(4-chloro-2-(methylsulfonyl)pyrimidine-5-yl)cyclopropane-1-carboxylate (2.6 g, 8.53 mmol) and benzyl 4-formamidopiperidine-1-carboxylate (2.350 g, 8.96 mmol) in THF (28.4 ml), sodium hydride (0.512 g, 12.80 mmol, 60% in mineral oil) was added. The reaction mixture was stirred at 100°C for 1 hour, then cooled to room temperature and quenched with saturated ammonium chloride. The mixture was diluted with water and ethyl acetate. The organic layer was washed with water and brine, dried over sodium sulfate, and concentrated. The crude product was purified with Biotage® (0-100% ethyl acetate in hexa...

Claims

1. A pharmaceutical product for use in a method of treating a human subject having a disease or disorder related to cyclin-dependent kinase 2 (CDK2), comprising a CDK2 inhibitor, wherein the human subject is (i) (a) Having a nucleotide sequence that encodes the p16 protein, including the amino acid sequence of Sequence ID No. 1, (b) Having a cyclin-dependent kinase inhibitor 2A (CDKN2A) gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) expressing the p16 protein, and (ii) (a) Having amplification of the cyclin E1 (CCNE1) gene, and / or (b) It has been determined that the expression level of CCNE1 in the biological samples obtained from the human subjects is higher than the control expression level of CCNE1, where the control expression level of CCNE1 is (1) the expression level of CCNE1 in samples (may be multiple) obtained from one or more subjects that did not respond to treatment with the CDK2 inhibitor, or (2) the expression level of CCNE1 in COV504 cells. The aforementioned CDK2 inhibitor 4-((8-cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine-2-yl)amino)benzenesulfonamide, 8-Cyclopentyl-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-phenyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-(1,1-difluorobutan-2-yl)-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-8-((1-methyl-1H-pyrazole-5-yl)methyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-(tetrahydrofuran-3-yl)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-((1R,2R)-2-hydroxy-2-methylcyclopentyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrido[2,3-d]pyrimidine-7(8H)-one, Jinaciclib, Arbosiclib, Sericiclib, Ronisiclib, Milsicrib, Abemaciclib, Trilaciclib, 【Chemistry 1】 Or a pharmaceutically acceptable salt thereof, The disease or disorder associated with the aforementioned CDK2 is cancer. Pharmaceuticals.

2. The aforementioned human subject, (i) (a) Having a nucleotide sequence that encodes a p16 protein, including the amino acid sequence of Sequence ID No. 1, and / or (b) CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and (ii) The pharmaceutical product according to claim 1, wherein it has been determined to have amplification of the CCNE1 gene in a biological sample obtained from the human subject.

3. The method described above is (i) In the biological sample obtained from the human subject, (a) A nucleotide sequence encoding the p16 protein, including the amino acid sequence of Sequence ID No. 1, (b) A cyclin-dependent kinase inhibitor 2A (CDKN2A) gene lacking one or more inactivating nucleic acid substitutions, and / or (c) Identify the presence of the p16 protein. (ii) In the biological sample obtained from the human subject, (a) amplification of the cyclin E1 (CCNE1) gene, and / or (b) Identify CCNE1 expression levels higher than the control CCNE1 expression level, where the control CCNE1 expression level is (1) the CCNE1 expression level in a sample(s) obtained from one or more subjects that did not respond to treatment with the CDK2 inhibitor, or (2) the CCNE1 expression level in COV504 cells. Furthermore (iii) A CDK2 inhibitor for use according to claim 1, comprising administering the CDK2 inhibitor to the human subject.

4. The method described above is (i) In the biological sample obtained from the human subject, (a) A nucleotide sequence encoding the p16 protein, including the amino acid sequence of Sequence ID No. 1, (b) CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) Identify the presence of the p16 protein. (ii) In the biological sample obtained from the human subject, (a) Identifying the amplification of the CCNE1 gene, and (iii) The pharmaceutical product according to claim 3, comprising administering a CDK2 inhibitor to the human subject.

5. The pharmaceutical product according to claim 1 or 3, wherein the expression level of CCNE1 in the biological sample is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 50 times, at least 75 times, or at least 100 times higher than the control expression level of CCNE1.

6. The pharmaceutical product according to any one of claims 1 to 5, wherein the CDKN2A gene encodes a protein containing the amino acid sequence of SEQ ID NO:

1.

7. The pharmaceutical product according to any one of claims 1 to 6, wherein the second therapeutic agent is administered to the human subject in combination with the CDK2 inhibitor.

8. The pharmaceutical product according to claim 7, wherein the second therapeutic agent is a BCL2 inhibitor or a CDK4 / 6 inhibitor.

9. A method for predicting the response to a CDK2 inhibitor in human subjects with a disease or disorder related to cyclin-dependent kinase 2 (CDK2), wherein the method is (i) From the biological sample obtained from the human subject, (a) Nucleotide sequence of the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene, (b) The presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) To determine the presence of the p16 protein, and (ii) From the biological sample obtained from the human subject, (a) Copy number of the cyclin E1 (CCNE1) gene, and / or (b) including determining the expression level of CCNE1, （１） (a) The presence of the CDKN2A gene which encodes the p16 protein containing the amino acid sequence of Sequence ID No. 1, (b) The presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, and / or (c) The presence of the p16 protein, and (2) It is predicted that the human subject will respond to the CDK2 inhibitor by amplification of the CCNE1 gene and / or by an expression level of CCNE1 higher than the control expression level of CCNE1, where the control expression level of CCNE1 is (1) the expression level of CCNE1 in a sample(s) obtained from one or more subjects that did not respond to treatment with the CDK2 inhibitor, or (2) the expression level of CCNE1 in COV504 cells. The aforementioned CDK2 inhibitor 4-((8-cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine-2-yl)amino)benzenesulfonamide, 8-Cyclopentyl-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-phenyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-(1,1-difluorobutan-2-yl)-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-8-((1-methyl-1H-pyrazole-5-yl)methyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-(tetrahydrofuran-3-yl)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-((1R,2R)-2-hydroxy-2-methylcyclopentyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrido[2,3-d]pyrimidine-7(8H)-one, Jinaciclib, Arbosiclib, Sericiclib, Ronisiclib, Milsicrib, Abemaciclib, Trilaciclib, 【Chemistry 2】 Or a pharmaceutically acceptable salt thereof, The disease or disorder associated with the aforementioned CDK2 is cancer. The aforementioned method.

10. (i) From the biological sample obtained from the human subject, (a) Determining the presence of a CDKN2A gene lacking a nucleotide sequence and / or one or more inactivated nucleic acid substitutions and / or deletions, and (ii) From the biological sample obtained from the human subject, (a) including determining the copy number of the CCNE1 gene, （１） (a) The presence of the CDKN2A gene encoding the p16 protein containing the amino acid sequence of Sequence ID No. 1, and / or (b) The presence of the CDKN2A gene lacking one or more inactivated nucleic acid substitutions and / or deletions, (2) The method according to claim 9, wherein amplification of the CCNE1 gene is predicted to cause the human subject to respond to the CDK2 inhibitor.

11. (i) The amplification of the CCNE1 gene comprises at least 3, at least 5, or at least 21 gene copies, and / or (ii) The expression level of CCNE1 is equal to (a) the expression level of CCNE1 mRNA, or (b) The pharmaceutical product according to any one of claims 1 to 8, which is the expression level of the CCNE1 protein.

12. The pharmaceutical product according to any one of claims 1 to 8, wherein the expression level of CCNE1 is measured by RNA sequencing, quantitative polymerase chain reaction (PCR), in situ hybridization, nucleic acid array, or RNA sequencing, and the expression level of the CCNE1 protein is measured by Western blotting, enzyme-linked immunosorbent, or immunohistochemical staining.

13. (i) The amplification of the CCNE1 gene comprises at least 3, at least 5, or at least 21 gene copies, and / or (ii) The expression level of CCNE1 is equal to (a) the expression level of CCNE1 mRNA, or (b) The method according to claim 9 or 10, wherein the expression level of the CCNE1 protein.

14. The method according to claim 13, wherein the expression level of CCNE1 is measured by RNA sequencing, quantitative polymerase chain reaction (PCR), in situ hybridization, nucleic acid array, or RNA sequencing, and the expression level of the CCNE1 protein is measured by Western blotting, enzyme-linked immunosorbent, or immunohistochemical staining.

15. A method for evaluating the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene and the cyclin E1 (CCNE1) gene, comprising: (i) determining the presence of a CDKN2A gene lacking a nucleotide sequence or one or more inactivating nucleic acid substitutions and / or deletions, and (ii) determining the copy number of the CCNE1 gene, from a biological sample (may be multiple) obtained from a human subject having cancer, which is a disease or disorder associated with cyclin-dependent kinase 2 (CDK2).

16. A method for evaluating the response of human subjects with cyclin-dependent kinase 2 (CDK2) related diseases or disorders to a CDK2 inhibitor, wherein the human subjects have been determined to have amplification of the cyclin E1 (CCNE1) gene and / or a CCNE1 expression level higher than the control expression level of CCNE1, where the control expression level of CCNE1 is (1) the CCNE1 expression level in a sample(s) obtained from one or more subjects that did not respond to treatment with the CDK2 inhibitor, or (2) the CCNE1 expression level in COV504 cells. The method described above is (a) The procedure involves measuring the phosphorylation level of retinoblastoma (Rb) protein at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample obtained from the subject after administration of a CDK2 inhibitor, When compared to the control level of Rb phosphorylation at serine corresponding to amino acid position 780 of SEQ ID NO: 3, the decrease in the Rb phosphorylation level at serine corresponding to amino acid position 780 of SEQ ID NO: 3 indicates that the human subject responds to the CDK2 inhibitor. The aforementioned CDK2 inhibitor 4-((8-cyclopentyl-6,6-dimethyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidine-2-yl)amino)benzenesulfonamide, 8-Cyclopentyl-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-phenyl-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-(1,1-difluorobutan-2-yl)-6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-8-((1-methyl-1H-pyrazole-5-yl)methyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 6,6-dimethyl-2-((1-(methylsulfonyl)piperidine-4-yl)amino)-8-(tetrahydrofuran-3-yl)-5,8-dihydropyrido[2,3-d]pyrimidine-7(6H)-one, 8-((1R,2R)-2-hydroxy-2-methylcyclopentyl)-2-((1-(methylsulfonyl)piperidine-4-yl)amino)pyrido[2,3-d]pyrimidine-7(8H)-one, Jinaciclib, Arbosiclib, Sericiclib, Ronisiclib, Milsicrib, Abemaciclib, Trilaciclib, 【Transformation 3】 Or a pharmaceutically acceptable salt thereof, The disease or disorder associated with the aforementioned CDK2 is cancer. The aforementioned method.

17. A method for measuring the amount of phosphorylation of retinoblastoma (Rb) protein at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in a biological sample, (a) To provide biological samples obtained from human subjects having diseases or disorders related to cyclin-dependent kinase 2 (CDK2), and (b) The method comprising measuring the phosphorylation level of the Rb protein at serine corresponding to amino acid position 780 of SEQ ID NO: 3 in the biological sample.

18. The method according to claim 16 or 17, wherein the biological sample includes a blood sample or a tumor biopsy sample.

19. (i) The cancer is a squamous cell carcinoma of the lung, adenocarcinoma of the lung, adenocarcinoma of the pancreas, invasive breast cancer, carcinosarcoma of the uterus, serous cystadenocarcinoma of the ovary, adenocarcinoma of the stomach, esophageal cancer, urothelial carcinoma of the bladder, mesothelioma, or sarcoma. (ii) The cancer is lung adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, or gastric adenocarcinoma. (iii) The cancer is an adenocarcinoma, carcinoma, or cystadenomatous carcinoma. (iv) The cancer is uterine cancer, ovarian cancer, stomach cancer, esophageal cancer, lung cancer, bladder cancer, pancreatic cancer, or breast cancer. (v) The cancer is ovarian cancer, uterine carcinosarcoma, or breast cancer. (vi) The cancer contains p27 inactivation, or (vii) The pharmaceutical product according to any one of claims 1 to 8, 11 and 12, wherein the cancer is N-myc-amplified neuroblastoma, K-Ras mutant lung cancer, or cancer having an FBW7 mutation and CCNE1 overexpression.

20. (i) The cancer is a squamous cell carcinoma of the lung, adenocarcinoma of the lung, adenocarcinoma of the pancreas, invasive breast cancer, carcinosarcoma of the uterus, serous cystadenocarcinoma of the ovary, adenocarcinoma of the stomach, esophageal cancer, urothelial carcinoma of the bladder, mesothelioma, or sarcoma. (ii) The cancer is lung adenocarcinoma, invasive breast cancer, uterine carcinosarcoma, ovarian serous cystadenocarcinoma, or gastric adenocarcinoma. (iii) The cancer is an adenocarcinoma, carcinoma, or cystadenomatous carcinoma. (iv) The cancer is uterine cancer, ovarian cancer, stomach cancer, esophageal cancer, lung cancer, bladder cancer, pancreatic cancer, or breast cancer. (v) The cancer is ovarian cancer, uterine carcinosarcoma, or breast cancer. (vi) The cancer contains p27 inactivation, or (vii) The method according to any one of claims 9, 10, 13, and 14, wherein the cancer is N-myc-amplified neuroblastoma, K-Ras mutant lung cancer, or cancer having an FBW7 mutation and CCNE1 overexpression.

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