Crystallographic forms and uses of checkpoint kinase 1 (CHK1) inhibitors

Abstract

This specification provides a pharmaceutical composition (1) comprising a CHK1 inhibitor.

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Application No. 63 / 493,846, filed on April 3, 2023, which is hereby incorporated by reference in its entirety.

Background Art

[0002] Cancer remains the second leading cause of death in the United States (US), accounting for approximately 1.9 million new diagnoses and 610,000 deaths on an annual basis. Worldwide, cancer is the second leading cause of death, causing nearly 10 million deaths in 2020, with nearly 1 in 6 deaths being due to cancer. The number of new cases is predicted to increase by 70% over the next 20 years.

[0003] In solid malignancies, metastatic spread and systemic disease account for approximately 90% of cancer-related deaths. Despite progress over the past several decades, the development of targeted interventions for advanced or metastatic solid tumors remains needed.

[0004] Patients with cancers having oncogene amplifications other than HER2 (or ERBB2) do not have approved targeted therapies as standard care. In general, these patients have worse survival rates than patients with other forms of oncogene changes or patients without known oncogene changes.

[0005] Currently, there are no approved treatments for patients with these other oncogene amplified tumors, particularly oral treatments containing a stable pharmaceutical form of a CHK1 inhibitor.

Summary of the Invention

[0006] Herein, 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridin-3-yl)-1H-pyrazol-3-yl)amino)pyrazine-2-carbonitrile:

[0007] [ka] Or, a crystalline form of a pharmaceutically acceptable salt or solvate thereof is disclosed.

[0008] In this specification, the free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0009] [ka] Alternatively, the crystalline form of the pharmaceutically acceptable solvate thereof is also disclosed.

[0010] In this specification, the anhydrous free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0011] [ka] The crystalline form of the crystals is also disclosed.

[0012] In some embodiments, crystalline compound 1 is a free base form FB-1 characterized by having at least one of the following properties: (a) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 1, (b) X-ray powder diffraction (XRPD) patterns with characteristic peaks at 11.96±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, and 26.75±0.1°2θ. (c) X-ray powder diffraction (XRPD) pattern with characteristic peaks at 13.87±0.1°2θ and 17.06±0.1°2θ. (d) A differential scanning calorimetry (DSC) thermogram with endothermic properties and a peak temperature at approximately 216.5°C (start). (e) A thermogravimetric thermal analysis (TGA) thermogram that is substantially the same as the one shown in Figure 2. (f) A thermogravimetric analysis (TGA) thermogram showing a mass loss of approximately 0.10% from the start of heating up to approximately 100.0°C, or (g) combinations of those.

[0013] This specification also discloses pharmaceutical compositions comprising the crystalline forms and pharmaceutically acceptable excipients disclosed herein.

[0014] This specification also discloses methods for treating cancer in subjects requiring treatment, including administering the crystalline forms disclosed herein to the subject.

[0015] In some embodiments, the cancer includes solid tumors.

[0016] In some embodiments, the cancer includes locally advanced or metastatic, unresectable solid tumors.

[0017] In some embodiments, the cancer includes a tumor or tumor cells having oncogene amplification.

[0018] In some embodiments, the oncogene amplification includes amplification of ABL, AKT1, AKT2, ALK, androgen receptor, BRAF, CCND1, CCND2, CCND3, CCNE1, CDK12, CDK4, CDK6, EGFR, ERBB2, EZH2, FGFR1, FGFR2, FGFR3, FLT3, IDH1 / 2, JAK2, JAK3, KIT, KRAS, MDM2, MDM4, MET, MYC, MYCL, MYCN, NRAS, PDGFRA, TERT, VEGFRA, or any combination thereof.

[0019] In some embodiments, the cancer gene amplification is present on ecDNA.

[0020] In some embodiments, the cancer gene amplification is present on one or more chromosomal loci.

[0021] In some embodiments, the cancer gene amplification is an amplification derived from ecDNA.

[0022] In some embodiments, the cancer is ovarian cancer.

[0023] In some embodiments, the ovarian cancer is platinum-resistant high-grade serous ovarian cancer, primary peritoneal cancer, or fallopian tube cancer.

[0024] In some embodiments, the cancer is uterine cancer.

[0025] In some embodiments, the uterine cancer is high-grade endometrial cancer, uterine serous carcinoma, or carcinosarcoma of the uterus.

[0026] In some embodiments, the cancer is colorectal cancer, esophageal cancer, gastric cancer, gastroesophageal junction (GEJ) cancer, head and neck squamous cell carcinoma, liposarcoma, non-small cell lung cancer, or subtype squamous cell carcinoma.

[0027] In some embodiments, the cancer is neuroblastoma.

[0028] In some embodiments, the cancer is breast cancer, cholangiocarcinoma, esophageal cancer, cervical squamous cell carcinoma, non-small cell lung cancer, gastric cancer, or subtype squamous cell carcinoma.

[0029] In some embodiments, the cancer is esophageal cancer, non-small cell lung cancer, sarcoma, or gastric cancer.

[0030] In some embodiments, the treatment further comprises administering an additional therapeutic agent.

[0031] In some embodiments, the oncogene amplification includes CDK4, CDK6, EGFR, FGFR1, FGFR2, or FGFR3.

[0032] In some embodiments, the treatment further includes administering a CDK4 / 6 inhibitor, an EGFR inhibitor, or an FGFR inhibitor. In some embodiments, the EGFR inhibitor is erlotinib. In some embodiments, the FGFR inhibitor is pemigatinib. In some embodiments, the FGFR inhibitor is futivatinib. In some embodiments, the CDK4 / 6 inhibitor is abemaciclib.

[0033] Reference All publications, patents, and patent applications referenced herein are incorporated herein by reference for the specific purposes identified herein. [Brief explanation of the drawing]

[0034] Novel features of the present invention are specifically described in the appended claims. For a better understanding of the features and advantages of the present invention, please refer to the following detailed description and the appended drawings, which describe exemplary embodiments in which the principles of the present invention are used. [Figure 1] This figure shows the X-ray powder diffraction (XRPD) pattern of the free base form FB-1 of compound 1. [Figure 2] This figure shows the thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram of the free base form FB-1 of compound 1. [Figure 3] This figure shows the X-ray powder diffraction (XRPD) pattern of the free base form FB-2 of compound 1. [Figure 4] This figure shows the X-ray powder diffraction (XRPD) pattern of the free base form FB-3 of compound 1. [Figure 5] This figure shows the X-ray powder diffraction (XRPD) pattern of the free base form FB-4 of compound 1. [Figure 6] This figure shows the X-ray powder diffraction (XRPD) pattern of the free base form FB-5 of compound 1. [Modes for carrying out the invention]

[0035] High-copy-number localized oncogene amplification is frequently observed to occur on extrachromosomal DNA (ecDNA). ecDNA is found in tumor cells and originates from extrachromosomal fragments of genomic DNA, often encoding full-length genes and regulatory regions such as promoters. ecDNA can appear physically distinct from chromosomes and possesses unique properties, including an open chromatin structure associated with overtranscription and a tendency to undergo structural mutations. In addition, because ecDNA lacks a centromere, its extrachromosomal location allows for inheritance during cell division via non-Mendelian segregation without kinetochores, enabling high-copy-number genetic heterogeneity across the tumor cell population. Due to these properties, ecDNA is a common cellular mechanism for oncogene amplification (e.g., EGFR), promoting overtranscription and overexpression of oncogenic proteins that lead to tumor growth and survival. Furthermore, these properties result in unparalleled genomic plasticity in tumor cells with amplified oncogenes via ecDNA, facilitating both rapid genomic evolution and avoidance of both carcinogenesis and treatment pressures. Cancer cells with oncogene amplification on ecDNA experience high levels of endogenous DNA replication stress (RS). Checkpoint kinase 1 (CHK1) plays a crucial role in managing RS; therefore, CHK1 is a potential therapeutic target for cancers with high endogenous RS, including those with oncogene amplification via ecDNA. Consistent with this hypothesis, ecDNA-amplified tumor cells are more sensitive to CHK1 inhibition than ecDNA-negative, non-amplified cells. When targeted therapy (e.g., EGFR inhibitors) is applied to the protein product of oncogenes amplified on ecDNA (e.g., EGFR), cancer cells are induced to evade such pressure, and these resistance mechanisms further increase RS and their dependence on CHK1. Therefore, combining targeted therapy pressure (e.g., EGFR inhibitors) with CHK1 pressure (i.e., CHK1 inhibitors) in ecDNA-amplified tumor cells yields a synergistic therapeutic effect.Therefore, as recognized and addressed herein, there is a need to provide CHK1 inhibitors with desired clinical and therapeutic properties, including stable pharmaceutical forms of such CHK1 inhibitors.

[0036] definition The following description includes specific details to fully understand the various embodiments. However, those skilled in the art will understand that the invention can be carried out without these details. In other examples, well-known structures are not illustrated or described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless otherwise required by context, the word “comprise,” and its variations such as “comprises” and “comprising,” should be interpreted in an open and comprehensive sense, i.e., “including, but not limited,” throughout the following specification and claims. Furthermore, the headings provided herein are for convenience only and do not constitute an interpretation of the scope or meaning of the claimed invention.

[0037] Throughout this specification, any reference to “some embodiments” or “an embodiment” means that a particular characteristic, structure, or feature described in relation to an embodiment is included in at least one embodiment. Therefore, where the phrases “in one embodiment” or “in an embodiment” appear in various places throughout this specification, they do not necessarily all refer to the same embodiment. Furthermore, a particular characteristic, structure, or feature may be combined in any preferred manner in one or more embodiments. Also, where used in this specification and the appended claims, the singular forms “a,” “an,” and “the” refer to multiple subjects unless otherwise clearly indicated. And note that the term “or” is generally used to mean “and / or” unless otherwise clearly indicated.

[0038] The terms “treat,” “treated,” “treatment,” and “treating,” as used herein, refer to therapeutic treatments whose purpose is to prevent or slow (reduce) an undesirable physiological disease, disorder, or illness, or to obtain beneficial or desired clinical outcomes. Beneficial or desired clinical outcomes for the purposes described herein include, but are not limited to, symptom reduction, a reduction in the severity of a disease, disorder, or illness, stabilization (i.e., no worsening) of the disease, disorder, or illness, delay in the onset or slowing of the progression of the disease, disorder, or illness, improvement of the disease, disorder, or illness, and remission, or improvement or improvement of the disease, disorder, or illness, whether detectable or undetectable (whether partially or whole). Treatment includes inducing a clinically significant response without excessive levels of side effects. Treatment also includes extended survival compared to survival expected without treatment. The terms “treat,” “treated,” “treatment,” or “treating,” as used herein, do not necessarily imply 100% or complete treatment. Rather, the degree of treatment that a person skilled in the art would recognize as having potential benefits or therapeutic effects varies. In this regard, the methods disclosed can provide treatment of any amount or level of impairment in mammals. For example, impairment, including its symptoms or disease, can be reduced by, for example, about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10%.

[0039] The terms “effective dose” or “therapeutic effective dose,” as used herein, refer to a sufficient amount of the compound disclosed herein administered to alleviate, to some extent, one or more of the symptoms of the disease or illness being treated, for example, cancer or inflammatory disease. In some embodiments, this results in a reduction and / or mitigation of the signs, symptoms, or causes of the disease, or any other desired change in the biological system. For example, an “effective dose” for therapeutic use is the amount of a composition containing the compound disclosed herein required to clinically significantly reduce the symptoms of the disease. In some embodiments, the appropriate “effective” dose in any individual case is determined using techniques such as dose escalation studies.

[0040] As used herein, the term "biological sample" generally refers to a sample derived from or obtained from a subject such as a mammal (e.g., human). Biological samples include, but are not limited to, hair, fingernails, skin, sweat, tears, eye fluid, nasal swabs or nasopharyngeal lavage fluid, sputum, pharyngeal swabs, saliva, mucus, blood, serum, plasma, placental fluid, amniotic fluid, umbilical cord blood, emphatic fluids, cavity fluid, earwax, oil, glandular secretions, bile, lymph, pus, microbiome, meconium, breast milk, bone marrow, bone, CNS tissue, cerebrospinal fluid, adipose tissue, synovial fluid, feces, gastric juice, urine, semen, vaginal secretions, stomach, small intestine, large intestine, rectum, pancreas, liver, kidney, bladder, lungs, and other tissues and fluids derived from or obtained from a subject.

[0041] The terms “tumor” or “tumor cells,” as used herein, generally refer to cells that proliferate and divide excessively or fail to die when they should. In some cases, tumor cells are present in solid masses such as solid tumors, or in some cases, tumor cells are found in non-solid forms such as hematological malignancies. Tumors or tumor cells may also contain metastatic or metastatic cells, and cancer cells can separate from the original (initial) tumor and form new tumors in other organs or tissues of the body.

[0042] When used herein, the term "ecDNA signature" generally refers to one or more features common to tumors or tumor cells that are ecDNA+. In some cases, an ecDNA signature is selected from the group consisting of gene amplification, p53 loss-of-function mutations, absence of microsatellite instability (MSI-H), low levels of PD-L1 expression, low levels of tumor inflammation signature (TIS), low levels of tumor mutational burden (TMB), increased frequency of allele substitutions, insertions, or deletions (indels), and any combination thereof. In some cases, an ecDNA signature may include copy number increases (gene amplification) in conjunction with specific structural mutations. In some cases, an ecDNA signature may include localized amplification. In some cases, an ecDNA signature may include detection or identification of ecDNA using imaging techniques. In some cases, an ecDNA signature may not include any imaging or direct detection of ecDNA.

[0043] compound This specification describes methods for treating cancer in patients requiring treatment, including the administration of CHK1 inhibitors.

[0044] compound 1 In some embodiments, the CHK1 inhibitor is compound 1 or a pharmaceutically acceptable salt thereof. Compound 1 is 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0045] [ka] In some embodiments, compound 1 is a free base. In some embodiments, compound 1 is in the form of a salt.

[0046] Crystalline compound 1 In this specification, 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0047] [ka] Or, a crystalline form of a pharmaceutically acceptable salt or solvate thereof is disclosed.

[0048] In this specification, the free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0049] [ka] Alternatively, the crystalline form of the pharmaceutically acceptable solvate thereof is also disclosed.

[0050] In this specification, the anhydrous free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile:

[0051] [ka] The crystalline form of the crystals is also disclosed.

[0052] This specification also discloses the crystalline form of the HCl salt of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0053] This specification also discloses the crystalline form of the HCl salt of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (compound 1).

[0054] This specification also discloses the crystalline form of the maleate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0055] The crystalline form of the maleate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) is also disclosed herein.

[0056] This specification also discloses the crystalline form of the fumarate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0057] The crystalline form of the fumarate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) is also disclosed herein.

[0058] This specification also discloses the crystalline form of the citrate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0059] This specification also discloses the crystalline form of the citrate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1).

[0060] This specification also discloses the crystalline form of the lactate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0061] The crystalline form of the lactate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) is also disclosed herein.

[0062] This specification also discloses the crystalline form of hippurate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0063] This specification also discloses the crystalline form of hippurate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1).

[0064] This specification also discloses the crystalline form of the sulfate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1) or a pharmaceutically acceptable solvate thereof.

[0065] This specification also discloses the crystalline form of the sulfate of 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile (Compound 1).

[0066] Free base crystalline compound 1 form FB-1 In some embodiments, crystalline compound 1 is a free base form FB-1 characterized by having at least one of the following properties: (a) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 1, (b) X-ray powder diffraction (XRPD) patterns with characteristic peaks at 11.96±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, and 26.75±0.1°2θ. (c) X-ray powder diffraction (XRPD) pattern with characteristic peaks at 13.87±0.1°2θ and 17.06±0.1°2θ. (d) Differential scanning calorimetry (DSC) thermogram with endothermic properties, with a peak temperature at approximately 216.5°C (start). (e) A thermogravimetric analysis (TGA) thermogram that is substantially the same as the one shown in Figure 2. (f) A thermogravimetric analysis (TGA) thermogram showing a mass loss of approximately 0.10% from the start of heating up to approximately 100.0°C, or (g) combinations of those.

[0067] In some embodiments, the crystalline morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 1.

[0068] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 1.

[0069] In some embodiments, compound 1, i.e., morphology FB-1, of the crystalline free base has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 11.96±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, and 26.75±0.1°2θ.

[0070] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 13.87±0.1°²θ and 17.06±0.1°²θ.

[0071] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.87 ± 0.1°²θ.

[0072] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 17.06 ± 0.1°²θ.

[0073] In some embodiments, the X-ray powder diffraction (XRPD) pattern further includes a peak at 5.27 ± 0.1°²θ.

[0074] In some embodiments, the X-ray powder diffraction (XRPD) pattern further includes a peak at 13.34 ± 0.1°²θ.

[0075] In some embodiments, the X-ray powder diffraction (XRPD) pattern further includes a peak at 15.66 ± 0.1°²θ.

[0076] In some embodiments, the X-ray powder diffraction (XRPD) pattern further includes a peak at 22.24 ± 0.1°²θ.

[0077] In some embodiments, the X-ray powder diffraction (XRPD) pattern further includes a peak at 27.84 ± 0.1°²θ.

[0078] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 5.27 ± 0.1°²θ.

[0079] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 11.96 ± 0.1°²θ.

[0080] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.34 ± 0.1°²θ.

[0081] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.87 ± 0.1°²θ.

[0082] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 15.66 ± 0.1°²θ.

[0083] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 17.06 ± 0.1°²θ.

[0084] In some embodiments, compound 1 of the crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 18.81 ± 0.1°²θ.

[0085] In some embodiments, compound 1 of the crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 19.91 ± 0.1°²θ.

[0086] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 21.67 ± 0.1°²θ.

[0087] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 22.24 ± 0.1°²θ.

[0088] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 26.75 ± 0.1°²θ.

[0089] In some embodiments, compound 1 of the crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 27.84 ± 0.1°²θ.

[0090] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 15.66±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0091] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least two characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0092] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least three characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0093] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least four characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0094] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least five characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0095] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least six characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0096] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least seven characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0097] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least eight characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0098] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least nine characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0099] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least 10 characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0100] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has an X-ray powder diffraction (XRPD) pattern with at least 11 characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

[0101] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has a differential scanning calorimetry (DSC) thermogram with endothermic properties, with a peak temperature at approximately 216.5°C (start).

[0102] In some embodiments, compound 1, i.e., morphology FB-1, of the crystalline free base has substantially the same thermogravimetric (TGA) thermogram as shown in Figure 2.

[0103] In some embodiments, compound 1, i.e., morphology FB-1, of a crystalline free base, has a thermogravimetric analysis (TGA) thermogram showing a mass loss of about 0.10% from the start of heating up to about 100.0°C.

[0104] In some embodiments, compound 1, i.e., form FB-1, of a crystalline free base is physically and chemically stable.

[0105] In some embodiments, compound 1, i.e., form FB-1, of the crystalline free base is chemically stable.

[0106] In some embodiments, compound 1, i.e., form FB-1, of a crystalline free base, is chemically and physically stable according to a 4-week stability test in open and closed dishes at 25°C / 60%RH.

[0107] In some embodiments, compound 1, i.e., form FB-1, of a crystalline free base, is chemically and physically stable according to a 4-week stability test in open and closed dishes at 40°C / 75%RH.

[0108] In some embodiments, compound 1, i.e., form FB-1, of a crystalline free base, is chemically and physically stable according to a 4-week stability test in open and closed dishes at 60°C.

[0109] In some embodiments, compound 1, i.e., form FB-1, of a crystalline free base is highly crystalline.

[0110] In some embodiments, compound 1, i.e., form FB-1, of the crystalline free base has a high melting point.

[0111] In some embodiments, compound 1, i.e., form FB-1, of the crystalline free base is non-hygroscopic.

[0112] In some embodiments, compound 1, i.e., form FB-1, of the crystalline free base is anhydrous.

[0113] [Table 1]

[0114] Free base crystalline compound 1 form FB-2 In some embodiments, crystalline compound 1 is a free base form FB-2 characterized by having at least one of the following properties: (a) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 3, (b) X-ray powder diffraction (XRPD) patterns with characteristic peaks as shown in Table 2, or (c) combinations of those.

[0115] In some embodiments, the crystalline morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 3.

[0116] In some embodiments, compound 1, i.e., morphology FB-2, of the crystalline free base has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 2.

[0117] [Table 2]

[0118] Free base crystalline compound 1 form FB-3 In some embodiments, crystalline compound 1 is a free base form FB-3 characterized by having at least one of the following properties: (a) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 4, (b) X-ray powder diffraction (XRPD) patterns with characteristic peaks as shown in Table 3, or (c) combinations of those.

[0119] In some embodiments, the crystalline morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 4.

[0120] In some embodiments, compound 1, i.e., morphology FB-3, of the crystalline free base has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 3.

[0121] [Table 3]

[0122] Free base crystalline compound 1 form FB-4 In some embodiments, crystalline compound 1 is a free base form FB-4 characterized by having at least one of the following properties: (d) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 5, (e) X-ray powder diffraction (XRPD) patterns with characteristic peaks as shown in Table 4, or (f) combinations of those.

[0123] In some embodiments, the crystalline morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 5.

[0124] In some embodiments, compound 1, i.e., morphology FB-4, of the crystalline free base has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 4.

[0125] [Table 4]

[0126] Free base crystalline compound 1 form FB-5 In some embodiments, crystalline compound 1 is a free base form FB-5 characterized by having at least one of the following properties: (g) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 6, (h) X-ray powder diffraction (XRPD) patterns with characteristic peaks as shown in Table 5, or (i) A combination of those.

[0127] In some embodiments, the crystalline morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 6.

[0128] In some embodiments, compound 1, i.e., morphology FB-5, of the crystalline free base has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 5.

[0129] [Table 5]

[0130] Pharmaceutical composition In certain embodiments, the compounds described herein are administered as pure chemical substances. In some embodiments, the compounds described herein are combined with a pharmaceutically suitable or acceptable carrier (hereinafter also referred to herein as a pharmaceutically suitable (or acceptable) excipient, a physiologically suitable (or acceptable) excipient, or a physiologically suitable (or acceptable) carrier) selected based on a chosen route of administration and standard pharmacovigilance such as that described in Remington: The Science and Practice of Pharmacy (Gennaro, 21st edition, Mack Pub. Co., Easton, Pennsylvania (2005)).

[0131] Accordingly, this specification provides pharmaceutical compositions comprising a compound described herein, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, and a pharmaceutically acceptable excipient.

[0132] Pharmaceutical compositions are administered in a manner appropriate to the disease being treated (or prevented). The appropriate dose, as well as the preferred duration and frequency of administration, are determined by factors such as the patient's condition, the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. Generally, an appropriate dose and treatment regimen provides a composition in an amount sufficient to deliver therapeutic and / or preventive benefits (e.g., improved clinical outcomes such as increased overall response rate, increased duration of response, more frequent complete or partial remission, longer disease-free survival and / or overall survival, or reduced symptom severity). The optimal dose is generally determined using experimental models and / or clinical trials. The optimal dose depends on the patient's body mass, weight, or blood volume.

[0133] In some embodiments, the pharmaceutical composition is formulated for oral, topical (including oral and sublingual), rectal, vaginal, transdermal, parenteral, intrapulmonary, intradermal, intrathecal, epidural, or intranasal administration. Parenteral administration includes intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. In some embodiments, the pharmaceutical composition is formulated for intravenous injection, oral administration, inhalation, nasal administration, topical administration, or ophthalmic administration. In some embodiments, the pharmaceutical composition is formulated for oral administration. In some embodiments, the pharmaceutical composition is formulated for intravenous injection. In some embodiments, the pharmaceutical composition is formulated as tablets, pills, capsules, liquids, inhalants, nasal spray solvents, suppositories, suspensions, gels, colloids, dispersants, suspensions, solvents, emulsions, ointments, lotions, eye drops, or ear drops. In some embodiments, the pharmaceutical composition is formulated as tablets.

[0134] The preferred dose and dosage regimen are determined by conventional ranging techniques known to those skilled in the art. Generally, treatment is initiated with a dose less than the optimal dose of the compound disclosed herein. The dose is then gradually increased until the optimal effect is achieved under the given conditions.

[0135] method This specification discloses a method for treating cancer in a subject requiring treatment, comprising administering a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof to the subject. In some embodiments, the CHK1 inhibitor is compound 1 or a pharmaceutically acceptable salt thereof.

[0136] In some embodiments of the methods disclosed herein, the subject experiences a therapeutic response.

[0137] In some embodiments of the methods disclosed herein, the therapeutic response includes a reduction in the level of oncogene amplification in the tumor or tumor cells after treatment compared to the level of oncogene amplification in the tumor or tumor cells before treatment.

[0138] In some embodiments of the methods disclosed herein, the therapeutic response includes a reduction in one or more of the following compared to before treatment: tumor growth, tumor size, number of tumor cells, or tumor metastasis.

[0139] In some embodiments of the methods disclosed herein, the treatment response includes the treatment benefit. In some embodiments of the methods disclosed herein, the treatment benefit is stable disease (SD). In some embodiments of the methods disclosed herein, the treatment benefit is a partial response (PR). In some embodiments of the methods disclosed herein, the treatment benefit is a complete response (CR). In some embodiments of the methods disclosed herein, a complete response is determined by RECIST v1.1 (or RANO for GBM). In some embodiments of the methods disclosed herein, the treatment benefit is the duration of response (DOR). In some embodiments of the methods disclosed herein, the treatment benefit is progression-free survival (PFS). In some embodiments of the methods disclosed herein, the treatment benefit is overall survival (OS).

[0140] In some embodiments of the methods disclosed herein, cancer includes solid tumors.

[0141] In some embodiments of the methods disclosed herein, cancer includes locally advanced or metastatic unresectable solid tumors.

[0142] In some embodiments of the methods disclosed herein, cancer includes a tumor or tumor cells having oncogene amplification.

[0143] In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of ABL, AKT1, AKT2, ALK, androgen receptor, BRAF, CCND1, CCND2, CCND3, CCNE1, CDK12, CDK4, CDK6, EGFR, ERBB2, EZH2, FGFR1, FGFR2, FGFR3, FLT3, IDH1 / 2, JAK2, JAK3, KIT, KRAS, MDM2, MDM4, MET, MYC, MYCL, MYCN, NRAS, PDGFRA, TERT, VEGFRA, or any combination thereof.

[0144] In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of FGFR1, FGFR2, FGFR3, or a combination thereof. In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of FGFR1, FGFR2, FGFR3, or a combination thereof, and the treatment further includes administering an FGFR inhibitor.

[0145] In some embodiments of the methods disclosed herein, the oncogene amplification comprises CDK4, CDK6, EGFR, FGFR1, FGFR2, or FGFR3.

[0146] In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of EGFR. In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of EGFR, and the treatment further includes administering an EGFR inhibitor.

[0147] In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of CDK4, CDK6, or a combination thereof. In some embodiments of the methods disclosed herein, the oncogene amplification includes amplification of CDK4, CDK6, or a combination thereof, and the treatment further includes administering a CDK4 / 6 inhibitor.

[0148] In some embodiments of the methods disclosed herein, the oncogene amplification is located on ecDNA.

[0149] In some embodiments of the methods disclosed herein, the oncogene amplification is located on one or more chromosomal loci.

[0150] In some embodiments of the methods disclosed herein, the oncogene amplification is ecDNA-derived amplification.

[0151] In some embodiments of the methods disclosed herein, the oncogene amplification has a copy number of at least 6, at least 8, at least 10, at least 15, at least 20, or more than 20 copies of the oncogene or a portion thereof.

[0152] In some embodiments of the methods disclosed herein, the cancer includes malignant tumors whose symptoms are reduced, lessened, decreased, and / or completely cured by reducing its size, slowing or stopping its growth or spread, or by eliminating, suppressing, and / or inhibiting the function of CHK1. The target malignant tumors include, but are not limited to, head and neck cancer, gastrointestinal cancer (esophageal cancer, gastric cancer, duodenal cancer, liver cancer, biliary tract cancer (gallbladder cancer, bile duct cancer, etc.), pancreatic cancer, colorectal cancer (colon cancer, rectal cancer, etc.)), lung cancer (non-small cell lung cancer, small cell lung cancer, squamous cell lung cancer, lung adenocarcinoma, mesothelioma, etc.), breast cancer, reproductive organ cancer (ovarian cancer, uterine cancer, cervical cancer, endometrial cancer, etc.), urinary tract cancer (kidney cancer, bladder cancer, prostate cancer, testicular tumor, etc.), hematopoietic malignancies (leukemia, malignant lymphoma, multiple myeloma, etc.), bone and soft tissue tumors (e.g., soft tissue sarcoma, liposarcoma, and osteosarcoma), skin cancer, and brain tumors (e.g., glioblastoma).

[0153] In some embodiments of the methods disclosed herein, the term cancer is used in light of this disclosure according to its plain and ordinary meaning and refers to all types of cancer, neoplasms, or malignant tumors found in mammals, including leukemia, lymphoma, melanoma, neuroendocrine tumors, carcinomas, and sarcomas. Examples of cancers that can be treated with the compounds disclosed herein, or their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers or pharmaceutical compositions include acute myeloid leukemia, adrenocortical carcinoma, adrenal carcinoma, bladder cancer, bone cancer, brain cancer, breast cancer (e.g., ductal carcinoma, lobular carcinoma, primary, metastatic), breast cancer, endocrine cancer, hepatic stellate cell carcinoma, pancreatic stellate cell carcinoma, cervical cancer, colon cancer, colorectal cancer, ductal carcinoma, endometrial cancer, esophageal cancer, gastric cancer, urogenital tract cancer, glioblastoma, glioma, head and neck cancer, hepatocellular carcinoma, Hodgkin's disease, kidney cancer, leukemia (e.g., lymphoblastic leukemia, chronic lymphocytic leukemia, hairy cell leukemia), liver cancer (e.g., hepatocellular carcinoma), lobular carcinoma, lung cancer (e.g., non-small cell lung cancer, squamous cell lung carcinoma, adenocarcinoma, large cell lung cancer, small cell lung cancer, carcinoid Examples of malignant cancers include sarcomas, liposarcomas, lymph node cancers, lymphomas (e.g., mantle cell lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, marginal zone lymphoma, Burkitt lymphoma, non-Hodgkin lymphoma), malignant carcinoids, malignant hypercalcemia, malignant pancreatic insulinoma, medullary thyroid carcinoma, medulloblastoma, melanoma, mesothelioma, multiple myeloma, pancreatic endocrine or exocrine neoplasms, neuroblastoma, ovarian cancer, papillary Paget's disease, pancreatic cancer, papillary thyroid carcinoma, phyllodes tumors, precancerous skin lesions, primary thrombocytosis, prostate cancer (e.g., castration-resistant prostate cancer), renal cancer, rhabdomyosarcoma, salivary gland cancer, sarcomas, soft tissue sarcomas, squamous cell carcinoma (e.g., head, neck, or esophagus), gastric cancer, testicular cancer, thyroid cancer, bladder cancer, or uterine cancer. In an embodiment, the cancer is selected from bladder cancer, breast cancer, colon cancer, esophageal cancer, glioblastoma, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, salivary gland cancer, soft tissue sarcoma, pulmonary squamous cell carcinoma, gastric cancer, and uterine cancer.

[0154] In some embodiments of the methods disclosed herein, the cancer is ovarian cancer. In some embodiments of the methods disclosed herein, the ovarian cancer is platinum-resistant high-grade serous ovarian cancer, primary peritoneal cancer, or fallopian tube cancer.

[0155] In some embodiments of the methods disclosed herein, the cancer is uterine cancer. In some embodiments of the methods disclosed herein, the uterine cancer is high-grade endometrial cancer, serous uterine cancer, or uterine carcinosarcoma.

[0156] In some embodiments of the methods disclosed herein, the cancer is glioblastoma or neuroblastoma.

[0157] In some embodiments of the methods disclosed herein, the cancer is colorectal cancer, esophageal cancer, gastric cancer, gastroesophageal junction (GEJ) cancer, head and neck squamous cell carcinoma, liposarcoma, non-small cell lung cancer, or a subtype of squamous cell carcinoma.

[0158] In some embodiments of the methods disclosed herein, the cancer is breast cancer or head cancer, esophageal cancer, cervical squamous cell carcinoma, non-small cell lung cancer, gastric cancer, or a subtype of squamous cell carcinoma.

[0159] In some embodiments of the methods disclosed herein, the cancer is esophageal cancer, non-small cell lung cancer, sarcoma, or gastric cancer. In some embodiments of the methods disclosed herein, the cancer is esophageal cancer. In some embodiments of the methods disclosed herein, the cancer is non-small cell lung cancer. In some embodiments of the methods disclosed herein, the cancer is sarcoma. In some embodiments of the methods disclosed herein, the cancer is gastric cancer.

[0160] In some embodiments of the methods disclosed herein, the subject has received one or more prior treatments.

[0161] In some embodiments of the methods disclosed herein, the subject was unresponsive to one or more prior treatments.

[0162] In some embodiments of the methods disclosed herein, the subject developed resistance to one or more prior treatments.

[0163] In some embodiments of the methods disclosed herein, the one or more pretreatments are chemotherapy.

[0164] In some embodiments of the methods disclosed herein, the one or more prior treatments are PD1 antibodies.

[0165] In some embodiments of the methods disclosed herein, the one or more prior treatments are PD-L1 antibodies.

[0166] In some embodiments of the methods disclosed herein, one or more prior treatments are CTLA4 checkpoint inhibitors.

[0167] In some embodiments of the methods disclosed herein, one or more prior treatments are VEGF-targeted therapies (e.g., bevacizumab for ovarian cancer).

[0168] This specification discloses a method for treating cancer in a subject requiring treatment, comprising administering a pharmaceutical composition containing compound 1 or a pharmaceutically acceptable salt thereof to the subject, the method further comprising obtaining a diagnostic indicator of oncogene amplification in a biological sample from the subject.

[0169] In some embodiments of the methods disclosed herein, the diagnostic indicator is obtained before a first administration of compound 1 or a pharmaceutically acceptable salt thereof.

[0170] In some embodiments of the methods disclosed herein, the diagnostic indicator is obtained after a first administration of compound 1 or a pharmaceutically acceptable salt thereof.

[0171] In some embodiments of the methods disclosed herein, the diagnostic indicator is obtained after multiple administrations of compound 1 or a pharmaceutically acceptable salt thereof.

[0172] In some embodiments of the methods disclosed herein, the diagnostic index arises from a next-generation sequencing (NGS) based assay.

[0173] In some embodiments of the methods disclosed herein, the diagnostic index is derived from a fluorescence in situ hybridization (FISH) assay.

[0174] In some embodiments of the methods disclosed herein, the diagnostic indicator includes an indicator of oncogene amplification derived from ecDNA.

[0175] In some embodiments of the methods disclosed herein, the diagnostic index is obtained from a tumor or a fluid biopsy.

[0176] In some embodiments of the methods disclosed herein, the methods further include evaluating a sample from a subject for the presence or level of one or more of the following: gene amplification, localized gene amplification, ecDNA, HSR, or ecDNA signature.

[0177] In some embodiments of the methods disclosed herein, the methods further include obtaining information about the presence or level of one or more gene amplifications, localized gene amplifications, ecDNA, HSRs, or ecDNA signatures in tumors or tumor cells from a subject before, during, or after administration of compound 1 or a pharmaceutically acceptable salt thereof.

[0178] Oncogene amplification Oncogene amplification-associated tumors represent a segment of the cancer population with extremely high unmet needs. Patients whose cancers have high-copy oncogene amplification have significantly worse survival rates compared to the broader cancer population. A pan-cancer analysis of oncogene amplification tumors, cross-referenced with surveillance, epidemiology, and end results program data, shows that in the United States alone, this population represents more than 400,000 newly diagnosed cancer patients annually across multiple tumor types.

[0179] Despite the enormous health benefits and improved survival offered by precision medicine and targeted therapies for cancer, these treatments have unfortunately proven largely ineffective in oncogene-amplifying populations. In addition, immune checkpoint inhibitors (e.g., pembrolizumab) may not function well in oncogene-amplifying cancer populations, and excessive progression is associated with the oncogene-amplifying tumor setting.

[0180] To date, HER2 inhibitors (e.g., trastuzumab) for HER2-overexpressing breast cancer, gastroesophageal junction cancer, and gastric cancer are the only approved targeted therapies in cancer populations with oncogene amplification (or overexpression), and breast cancer is the only monotherapy approved. Targeted therapies that have shown efficacy in patients whose cancer is caused by oncogene point mutations, gene fusions, or skipping deletions have generally failed to demonstrate strong efficacy in patients whose tumors are caused by oncogene amplification. This lack of approved therapies has occurred despite extensive clinical trials of targeted therapies in cancer populations with oncogene amplification, including EGFR inhibitors in EGFR-amplified glioblastoma multiforme, FGFR inhibitors in FGFR-amplified cancer, and CDK4 / 6 inhibitors in CDK4-amplified liposarcoma. These clinical data have led to the misconception that oncogene amplification may not be an relevant cancer driver. This erroneous conclusion has been reached despite extensive data to the contrary. The recurrent, localized copy number amplification and overexpression of established (or otherwise activated by mutation and / or gene fusion) oncogene drivers, as well as the antitumor effects of targeted inhibition (genetic and pharmacological) in short-term preclinical cancer models, establish amplification as the driver, even if targeted therapeutic approaches generally do not translate into long-term clinical benefits. However, the aforementioned discrepancies suggest that cancers caused by oncogene amplification are biologically distinct from other tumors and require a new therapeutic paradigm. Therefore, a better understanding of the biology of oncogene amplification is needed, with the aim of advancing novel therapeutic approaches, drug targets, and new molecular entities for this patient population with high unmet needs.

[0181] The role of extrachromosomal DNA in oncogene amplification Chromosomal instability and tumor heterogeneity have been suggested to be the cause of many targeted therapy failures. Consistent with this hypothesis, oncogene amplification is a consequence of prior or ongoing chromosomal instability resulting from numerical and / or structural changes in chromosomes, which can give rise to ecDNA. It has long been recognized that oncogenes can amplify not only on chromosomes but also on ecDNA, originally referred to as "double microchromosomes." However, the frequency, importance, and specific role of ecDNA in cancer biology were not well understood until recently.

[0182] Some of the most common driver oncogenes are encoded on ecDNA and can confer a selective advantage to cancer cells. These oncogenes amplified on ecDNA possess several characteristics that distinguish them from oncogene amplifications localized on chromosomes. 1. ecDNA lacks a centromere and therefore, in contrast to amplification localized on chromosomes, it segregates unevenly into daughter cells during cell division. This characteristic supports non-Mendelian inheritance patterns, allowing for extreme copy number changes in genes with relatively few cell divisions, resulting in widespread copy number heterogeneity and leading to adaptive and tumor evolution. 2.ecDNA is epigenetically dysregulated and contains accessible chromatin regions and gene regions that are often more actively expressed than genes located on chromosomes and are overtranscribed.

[0183] These characteristics distinguish ecDNA from other forms of oncogene amplification, promoting levels of genomic plasticity and adaptability beyond chromosomal amplification, allowing tumors to evade environmental damage, including targeted therapy pressures. Overcoming ecDNA-mediated adaptation in oncogene-amplified tumors requires novel therapeutic approaches that disrupt ecDNA function.

[0184] Oncogene amplification via ecDNA is a major driver of carcinogenesis and plays a significant role in causing tumor heterogeneity, allowing cancer cells to rapidly develop resistance to targeted oncogene therapy. ecDNA-mediated oncogene amplification has been observed in approximately half of all human cancer types, but rarely in normal cells. For example, ecDNA-mediated oncogene amplification can be found in about 14% of early cancer samples, and more than half of all high-copy-number oncogene amplifications (i.e., copy number 8 or higher) are present on ecDNA. Furthermore, many of the most aggressive tumor types contain the highest prevalence of ecDNA, including approximately 60% of glioblastoma multiforme and less than 50% of sarcomas.

[0185] Patients with cancers containing ecDNA experience significantly shorter survival times than patients with cancers caused by other molecular damage, even when the tumor type is controlled. These data strongly suggest that patients with cancers containing ecDNA require a new therapeutic paradigm to address this significant unmet need.

[0186] The role of extrachromosomal DNA in treatment resistance The unique properties of ecDNA, coupled with the remarkable genomic plasticity of tumors mediated by ecDNA, contribute to the aggressive nature of tumors and their ability to evade therapeutic pressures through rapid genomic evolution. The first demonstration of ecDNA-induced treatment resistance was shown in mouse cancer cell lines, where methotrexate treatment resulted in high amplification of dihydrofolate reductase (DHFR) on ecDNA, which disappeared upon methotrexate removal. Similar examples of DHFR ecDNA amplification have been repeated in multiple human cancer cell lines. Furthermore, amplification of drug efflux pump genes on ecDNA, including the ABC transporter family, has been observed to mediate resistance to various chemotherapies. An equivalent role of ecDNA in providing resistance to more modern targeted therapies is also well established. Avoidance of therapeutic response to the EGFR inhibitor erlotinib was facilitated by the rapid disappearance of populations with EGFRvIII amplification on ecDNA in patient-derived glioblastoma pleomorphism cells, coinciding with the emergence of a new cell population containing MDM2 amplification on ecDNA. This observed effect is consistent with a similar lack of response to EGFR inhibitors in the patients. Preclinical studies in gastric cancer cell lines containing FGFR2 amplified on ecDNA demonstrated that cellular resistance to the pan-FGFR inhibitor infigratinib may be triggered by an oncogene-dependent switch from FGFR2 amplification on ecDNA to a new, rapid amplification of EGFR on ecDNA. Notably, this dependency reversed to FGFR2 amplification on ecDNA under erlotinib-mediated EGFR inhibitory pressure. In each case, the initial cell population was affected by the respective targeted therapy, resulting in a short-lived antiproliferative effect lasting several weeks. Resistance to targeted therapy and regrowth occurred concurrently with the switch of amplified oncogenes on ecDNA. The rapid rate of amplification is characteristic of ecDNA and helps explain the resistance to endogenous targeted therapy in de novo oncogene amplification cancers.

[0187] Similarly, mutant oncogenes (e.g., BRAFV600E and KRASG12C) may amplify on ecDNA as a resistance mechanism to corresponding targeted therapies (e.g., BRAF / MEK inhibitors or KRAS inhibitors). For example, mutant BRAFV600E melanoma cell lines developed ecDNA amplification of BRAFV600E after exposure to BRAF / MEK dual inhibition. This phenomenon has also been demonstrated in clinical cases. In relation to this, numerous putative acquired resistance mechanisms to the KRASG12C inhibitor adagrasib have been reported, among which high levels of localized amplification of KRASG12C on ecDNA, confirmed in vitro and in vivo, conferred resistance to both adagrasib and sotrasib, which are clinically validated KRASG12C inhibitors.

[0188] In summary, these studies highlight remarkable genomic plasticity and a rapid increase in ecDNA-mediated oncogene amplification, enabling cancer cells to adapt quickly to therapeutic pressures. In conclusion, cancers caused by ecDNA-mediated oncogene amplification are biologically distinct from other oncogene-activated tumors and require a new therapeutic paradigm.

[0189] Medication / Administration This specification discloses a method for treating cancer in a subject requiring treatment, comprising administering a pharmaceutical composition containing compound 1 or a pharmaceutically acceptable salt thereof to the subject. In some embodiments, the pharmaceutical composition containing compound 1 comprises a crystalline form of compound 1, a crystalline free base form of compound 1, or a crystalline anhydrous free base form of compound 1. In some cases, such a form of compound 1 has an X-ray powder diffraction (XRPD) pattern, a differential scanning calorimetry (DSC) thermogram, a thermogravimetric analysis (TGA) thermogram, or any combination thereof as described herein.

[0190] Theoretical basis for the starting dose of compound 1 in humans GLP-compliant repeated-dose toxicity studies were conducted in rats and dogs every other day for a maximum exposure period of 29 days, following oral force-feeding. The main effects observed in both species were related to myelosuppression / depletion and gastrointestinal toxicity, which were considered to be the intended pharmacological effects of compound 1, directly related to CHK1 inhibition. Major changes associated with compound 1 were demonstrated to be partially or completely reversible in all tissues, and no major unexpected toxicity was identified.

[0191] In some embodiments of the methods disclosed herein, compound 1 is administered in doses ranging from about 10 mg to about 800 mg.

[0192] In some embodiments of the methods disclosed herein, compound 1 is administered in doses ranging from about 10 mg to about 400 mg.

[0193] In some embodiments of the methods disclosed herein, compound 1 is administered in doses ranging from about 10 mg to about 400 mg, thereby causing the subject to experience a therapeutic response.

[0194] In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 10 mg and about 20 mg, about 20 mg and about 40 mg, about 40 mg and about 80 mg, about 80 mg and about 120 mg, about 120 mg and about 160 mg, about 160 mg and about 200 mg, and about 200 mg and about 400 mg.

[0195] In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 20 to about 40 mg, about 40 to about 80 mg, about 80 to about 120 mg, about 120 mg to about 160 mg, or about 160 mg to about 200 mg.

[0196] In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 10 mg and about 20 mg, about 20 mg and about 40 mg, about 40 mg and about 80 mg, or about 80 mg and about 120 mg.

[0197] In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 10 mg and about 20 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 20 mg and about 40 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 40 mg and about 80 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 80 mg and about 120 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 120 mg and about 160 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 160 mg and about 200 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is between about 200 mg and about 400 mg.

[0198] In some embodiments of the methods disclosed herein, the dose of compound 1 is about 10 mg, about 20 mg, about 40 mg, about 80 mg, about 120 mg, about 160 mg, or about 200 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 10 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 15 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 20 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 25 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 30 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 35 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 40 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 45 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is about 50 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 55 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 60 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 65 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 70 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 75 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 80 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 85 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 90 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 95 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 100 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 105 mg.In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 110 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 115 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 120 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 125 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 130 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 135 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 140 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 145 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 150 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 155 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 160 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 165 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 170 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 175 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 180 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 185 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 190 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 195 mg. In some embodiments of the methods disclosed herein, the dose of compound 1 is approximately 200 mg.

[0199] In some embodiments of the methods disclosed herein, the composition is administered orally.

[0200] In some embodiments of the methods disclosed herein, the composition is administered parentally.

[0201] In some embodiments of the methods disclosed herein, the composition is administered daily.

[0202] In some embodiments of the methods disclosed herein, the composition is administered every other day.

[0203] In some embodiments of the methods disclosed herein, the composition is administered in cycles of day 1 and day 3, followed by a 4-day drug-free period.

[0204] In some embodiments of the methods disclosed herein, the composition is administered every three days or weekly.

[0205] In some embodiments of the methods disclosed herein, the composition is administered every three days.

[0206] In some embodiments of the methods disclosed herein, the composition is administered weekly.

[0207] In some embodiments of the methods disclosed herein, the composition is administered with drug-free intervals of 4 days, 4-7 days, 7 days, or 14 days.

[0208] combination This specification discloses a method for treating cancer in a subject requiring treatment, comprising administering a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof and additional therapeutic agents to the subject. In some embodiments, the CHK1 inhibitor is compound 1 or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition comprising compound 1 comprises a crystalline form of compound 1, a crystalline free base form of compound 1, or a crystalline anhydrous free base form of compound 1. In some cases, such a form of compound 1 has an X-ray powder diffraction (XRPD) pattern, a differential scanning calorimetry (DSC) thermogram, a thermogravimetric analysis (TGA) thermogram, or any combination thereof as described herein.

[0209] In some embodiments of the methods disclosed herein, the additional therapeutic agent is an EGFR inhibitor.

[0210] In some embodiments of the methods disclosed herein, the EGFR inhibitors are 602, 705, 707, avivertinib, ABX-900, afatinib, agerafenib (RXDX-105), alflutinib mesylate, amivantamab, APL-1898, ASK-120067, aumorertinib (almonertinib), BBT-176, BDTX-1535, BDTX-189, BEBT-109, befortinib mesylate, beitatini, BLU-701, BLU-945, BPI-361175, BPI-7711, BPI-D0316, C-005, CDP1, Cetuximab, CH-7233163, CK-101, CMAB-017, Dacomitinib, Depatuxizumab, Depatuxizumab-Mahodotin (ABT-414), DFP-17729, Docitinib, DS-2087, DZD-9008, E01001, E-10C, Epertinib, Epitinib (HMPL-813), Erlotinib, ES-072, FCN-411, FHND-9041, Flumonertinib, FWD-1509, GB-263, GC-1118A, Gefitinib, GMA-204, GR-14 01, Hemay-022, HLX-07, HS-627, I-010, Icotinib, Imugatuzumab, IN-A008, JMT-101, JRF-103, JS-111, JS-113, JZB-28, KN-023, KN-026, KP-673, Lapatinib, Lalotinib, Lazertinib, LL-191, LYN205, M1231, Maifatinib, Marizomib, Mobocertinib, MP-0274, MRG003, Naptinib Tosylate, Nazartinib, Necitumumab, Neptinib, Nimotuzumab, NRC-2694-A, NT- 004, OBX1-012, olafertinib, olmutinib, ORIC-114, olitinib, osimertinib, panitumumab, pirotinib, poziotinib, PRB-001, pyrotinib, QL-1203, SCT-200, cerculutamab, SHR-A1307, SIM-200, SPH-1188, SSGJ-612, SYN-004, TAD-011, tarloxotinib, TAS-6417, TGRX-360, teriatinib (HMPL-309), TPC-064, TQB-3804, TY-9591, WJ-13404, WSD-0922,The drug is selected from the group consisting of XZP-5809, yinlitinib maleate, YK-029A, YZJ-0318, zolifertinib, and ZSP-0391.

[0211] In some embodiments of the methods disclosed herein, the EGFR inhibitor is erlotinib. Erlotinib (TARCEVA®) is an oral small molecule inhibitor of the receptor tyrosine kinase EGFR. Early data have shown anticancer activity in several tumors, including lung and pancreatic cancers, and erlotinib was approved by the FDA in 2013 for EGFR mutant NSCLC with EGFR exon 19 deletion or exon 21 substitution mutations. In particular, erlotinib can inhibit wild-type EGFR and is not selective for mutant EGFR only.

[0212] The planned dose of erlotinib is 150 mg orally once daily, at least one hour before or two hours after food intake. This is the dose of erlotinib for the treatment of NSCLC according to the United States Prescribing Information (USPI) for TARCEVA®.

[0213] In some embodiments of the methods disclosed herein, erlotinib is administered to subjects at doses of 150 mg orally per day, 100 mg orally per day, or 50 mg orally per day.

[0214] In some embodiments of the methods disclosed herein, the additional therapeutic agent is an FGFR inhibitor.

[0215] In some embodiments of the methods disclosed herein, the FGFR inhibitors are 3D-185, ABSK-011, ABSK-012, ABSK-061, ABSK-091, Aldafermin, Allofanib, AST-56100, AZD-4547, Bemarituzumab, BFKB-8488A, BGS-2219, BIO-1262, BPI-17509, BPI-43487, CPL-304-110, Delazantinib, E-7090, Erdafitinib, EVER-4010001, EVT-601, FGF-401, Fisogati The following are selected from the group consisting of Nib, FPI-1966, Fuchibatinib, Gunagratinib, H3B-6527, HH-185, HMPL-453, HS-236, ICP-105, ICP-192, Infigratinib, JAB-6000, KIN-3248, M-6123, MAX-40279, OM-RCA-001, Pemigatinib, RLY-4008, Rogatinib, SAR-439115, SAR-442501, SC-0011, SY-4798, TT-00434, Zoligratinib (FF-284), and WXSH-0011.

[0216] In some embodiments of the methods disclosed herein, the FGFR inhibitor is pemigatinib. Pemigatinib (PEMAZYRE®) is an oral small molecule inhibitor of the receptor tyrosine kinase FGFR. Pemigatinib was first approved by the FDA in 2020 for previously treated, unresectable locally advanced or metastatic cholangiocarcinoma with FGFR2 fusion or other rearrangements detected by FDA-approved testing. In particular, pemigatinib can inhibit wild-type FGFR1, FGFR2, and FGFR3 receptors.

[0217] In some embodiments of the methods disclosed herein, pemigatinib is administered to subjects in doses of 13.5 mg orally per day, 9 mg orally per day, or 4.5 mg orally per day, once daily for 14 days, followed by a 7-day period without administration of pemigatinib.

[0218] In some embodiments of the methods disclosed herein, the FGFR inhibitor is futivatinib.

[0219] Futivatinib (LYTGOBI®) is indicated for the treatment of adults with previously treated, unresectable locally advanced or metastatic intrahepatic cholangiocarcinoma that has fibroblast growth factor receptor 2 (FGFR2) gene fusion or other rearrangement. Futivatinib was approved for medical use in the United States in September 2022.

[0220] In some embodiments of the methods disclosed herein, futivatinib is administered to subjects at an oral dose of 20 mg per day.

[0221] In some embodiments of the methods disclosed herein, the additional therapeutic agent is a CDK4 / 6 inhibitor.

[0222] In some embodiments of the methods disclosed herein, the CDK4 / 6 inhibitors include abemaciclib, AG-122275, AM-5992, AT-7519, AU2-94, auceliclib, BEBT-209, BPI-1178, BPI-16350, CS-3002, fascapricin, FCN-437, FN-1501, GLR-2007, GW-491619, HEC-80797, HS-10342, IIIM-290, IIIM-985, rerocyclib, milcyclib maleate, MM-D37K, MS-14 The following are selected from the group consisting of 0, NP-102, NUV-422, ON-123300, palbociclib, PF-06842874, PF-06873600, PF-07220060, QHRD-110, R-547, RGB-286199, RGT-419B, ribociclib, ribiciclib, RO-0505124, SHR-6390, THR-53, THR-79, TQB-3303, TQB-3616, trilaciclib, TY-302, TY-302, borsiclib, VS2-370, WXWH-0240, XH-30002, and XZP-3287.

[0223] In some embodiments of the methods disclosed herein, the CDK4 / 6 inhibitor is abemaciclib. Abemaciclib (VERZENIO®) is an oral small molecule inhibitor of CDK4 / 6. Abemaciclib was first approved by the FDA in 2017 for the treatment of hormone receptor-positive, HER2-negative advanced or metastatic breast cancer. In particular, abemaciclib can inhibit wild-type CDK4 and CDK6 receptors.

[0224] In some embodiments of the methods disclosed herein, abemaciclib is administered to subjects in doses of approximately 50 mg twice daily, approximately 100 mg twice daily, or 150 mg twice daily.

[0225] In some embodiments of the methods disclosed herein, the additional therapeutic agent is a BRAF inhibitor. In some embodiments of the methods disclosed herein, the BRAF inhibitor is ABM-1310, agerafenib (RXDX-105), ARQ-736, ASN-003, AZ-304, AZ-628, BAL-3833, berbarafenib, BGB-3245, BI-882370, dabrafenib, DAY101, DP-2874, EBI-907 These include EBI-945, Encorafenib, GDC-0879, Rifilafenib, LUT-014, LYN204, NMS-P285, NMS-P730, PF-04880594, PF-07284890, PLX-8394, RX-208, TL-241, UAI-201, UB-941, Vemurafenib, VS-6766, or XL-281.

[0226] In some embodiments of the methods disclosed herein, the additional therapeutic agent is an MDM2 or MDM4 inhibitor.

[0227] In some embodiments of the methods disclosed herein, the MDM2 inhibitor is AD-021.32, ALRN-6924, APG-115, ASTX-295, ATSP-7041, BI-907828, CGM-097, CYC700, DS-5272, Idasanutrin, KRT-232 (AMG-232), MD-224, MI-1061, MI-219, MI-43, MI-77301 (SAR405838, S These include AR299155), MK-8242, NU-8231, NVP-CGM097, OM-301, PXN-527, RAIN-32 (mirademethane), RG7112 (RO5045337), RG7388 (RG7775), Rigel-3, RO-2468, RO-5353, RO-5963, Seldemethane (JNJ-26854165), SIL-43, Silemadrin, or UBX-0101. In some embodiments of the methods disclosed herein, the MDM4 inhibitor is 17AAG, 489-PXN, ALRN-6924, APG-115, ATSP-7041, BI-907828, CTX1, FL-118, inulanolide A, K-178, or SAH-p53-8.

[0228] In some embodiments of the methods disclosed herein, the additional therapeutic agent is a MET inhibitor.

[0229] In some embodiments of the methods disclosed herein, the MET inhibitors are ABP-1130, BPI-1831, BPI-2021, BYON-3521, CG-203306, CX-1003, Debio-1144, EMD-94283, EMT-100, EMT-101, HE-003, LMV-12, LS-177, NX-125, OMO-2, and PF-4254. These include anti-MET antibodies such as 644, PRX-MET, PTX-2173, QBH-196, RP-1400, SAB-Y14, SAR-125844, SGX-126, SYD-3521, WXSH-0011, X-379, and XL-265, as well as ABX-900, GB-263, FS-101, LY-3164530, LY-3343544, PMC-002, or SAIT-301. In some embodiments of the methods disclosed herein, the MET inhibitors are ABN-401, ABT-700, AMG-208, AMG-337, ARGX-111, BAY-85-3474, BMS-817378, boditinib, BPI-9016M, gourmettinib, golbatinib tartrate, GST-HG161, HQP-8361, I-020, JNJ-38877605, canitinib, merestinib, MK-2461, MK-8033, OMO-1, pamfetinib, S-4 The anti-MET antibodies include 9076, savolitinib, SPH-3348, tivantinib, SAR-125844, SCR-1515, and TPX-0022, or APL-101, CKD-702, EMB-01, EMI-137, ficratuzumab, HLX-55, HS-10241, MCLA-129, MT-8633, NOV-1105, RC-108, REGN-5093, SHR-A1403, Sym-015, or terisotuzumab vedotin. In some embodiments of the methods disclosed herein, the MET inhibitor is amivantamab, capmatinib, crizotinib, or tepotinib.

[0230] In some embodiments of the methods disclosed herein, the additional therapeutic agent is a KRAS inhibitor. In some embodiments of the methods disclosed herein, the KRAS inhibitor is an anti-KRAS antibody such as ABREV01, ARS-1620, APG-1842, ATG-012, BBP-454, BEPT-607, BI-2852, BI-1823911, BPI-421286, BTX-2541, COTI-219, IMM-1811900, JAB-21000, JAB-22000, JAB-23000, JAB-BX300, JP-002, KR-12, LYN202, MRTX-1133, RAS-F, RMC-6236, RMC-6291, SDGR5, STX-301, and YL-15293, or SBT-100, SBT-102, or SBT-300. In some embodiments of the methods disclosed herein, the KRAS inhibitor is adagrasib, ARS-3248, D-1553, GDC-6036, JDQ-443, LY3537982, sotrasib (AMG510), or BI 1701963. [Examples]

[0231] Example 1: Characterization of the solid state of compound 1, form FB-1 Figure 1 shows the crystalline XRPD pattern of compound 1, morphology FB-1. DSC analysis at 216.6°C (T peak Endothermic effects were observed, with a loss of 0.11 wt% in the TGA between room temperature and 100°C (Figure 2).

[0232] [Table 6]

[0233] pKa measurement The material used in this test was FB-1. The dissociation constant of the free base of compound 1 was measured using potentiometric acid-base titration. The experimental procedure is described below. Three consecutive measurements suggested a pKa of 8.99 ± 0.6 (standard deviation). The experimental and predicted pKa values ​​were similar. procedure: 1) Approximately 10 mg of the starting material was weighed in 10 ml of MeOH. 2) 25 mL of 0.5 M NaCl was added. 3) The pH value was adjusted to approximately 3-4 with 0.1M HCl. 4) The above solution was titrated with 0.5 M NaOH solution from pH 3 to 12.

[0234] [Table 7]

[0235] Determination of water activity The material used in this test was FB-1. The water activity of the free base of compound 1 was measured using Aqualab TDL-2. The experimental procedure is described below. When the measurement was performed in two consecutive steps, the material showed a water activity of 0.47. procedure: 1) Approximately 3-5 mL of reference standard was placed in a sample dish. 2) Set the temperature of the Aqualab TDL-2 to 25°C. 3) Close the sample chamber and start the measurement. 4) Confirm that the values ​​of the reference solution are within the standard limits. 5) Take approximately 50-100 mg of API into a new sample dish and start the measurement.

[0236] [Table 8]

[0237] Determination of the partition coefficient by the shake flask method The Log P values ​​for the experiment were measured using the shake flask method, with the Log P in DI water being 2.01 ± 0.309 over 1 day (24 hours). Due to the low solubility of the compound 1 product in DI water, the slow steering method was chosen to continue the experiment, resulting in a Log P of 1.64 ± 0.259 over 5 days (120 hours) in DI water. Both the standard shake flask Log P procedure and the slow steering Log P procedure were used to determine the Log P. The average Log P results were obtained from three replicate tests using HPLC quantification. procedure: 1) The partition coefficient was determined under 1-octanol / aqueous phase conditions (the aqueous phase was deionized water, and the 1-octanol / aqueous phase ratio was 50:50). 2) Before determining the partition coefficients, the two solvents were mutually saturated at the experimental temperature. 500 mL of 1-octanol and 500 mL of aqueous phase were transferred to a 1 L bottle, shaken well for 24 hours, and then allowed to stand for at least 24 hours to allow the two phases to separate. 3) Test conditions - Approximately 10 mg of the sample was dissolved in 10 mL of saturated 1-octanol, and 9 mL of this was transferred to a 20 mL glass sample vial containing 9 mL of aqueous phase. Sample preparation was performed in three batches. 4) A 20 mL glass sample vial was vigorously shaken at 200 rpm for 24 hours using an orbital shaker (BT Lab Systems, model number BT909, Missouri, USA), and then the flask was allowed to stand for 2 hours to allow the system to reach equilibrium. This was a 1-day (24-hour) sample. Shaking was continued at 200 rpm for 96 hours, and then the flask was allowed to stand for 2 hours to allow the system to reach equilibrium. This was a 5-day (120-hour) sample. The solubility of compound 1 in water was too low to be determined using HPLC, and therefore the results of the 1-octanol layer were used for the calculation of Log P.

[0238] [Table 9]

[0239]

Table 10

[0240] Example 2: Polymorph Screening Solubility Test of FB-1 Estimate the approximate solubility of crystalline compound 1 in 20 solvents and report the situation / results in Table 11. Procedure: 1) Weighed approximately 5 mg of the starting material and placed it in a 4.0 mL vial. Added 25 μL of the solvent at room temperature. 2) Shook and stirred the solution. If a clear solution was not obtained, an additional 25 μL was added and repeated until 0.75 mL of the solvent was added. 3) Then, the solvent was added up to 3.0 mL in 0.25 mL increments. 4) Stirred the solution overnight.

[0241]

Table 11

[0242] Polymorph Screening of Form FB-1 of Compound 1 Based on the approximate solubility of the starting material, a total of 80 polymorph screening experiments were completed using various screening techniques, including poor solvent addition, solid vapor diffusion, liquid vapor diffusion, slurry at room temperature, slurry at 50 °C, slow evaporation, polymer-induced crystallization, grinding, heating·cooling·heating, and hydration experiments. The material used in this test was Form FB-1 of the free base compound 1. Procedure 1. Slurrying at Room Temperature and 50 °C Approximately 30 mg of the API was slurried in 0.5 - 1.0 mL of different solvents in a 4.0 mL glass vial at room temperature using a magnetic stirrer. The solids in the slurry were characterized by XRPD after 5 - 7 days. 2. Hydration at Room Temperature and 50 °C Approximately 30 mg of the API was slurried at room temperature in 0.5 - 1.0 mL of different solvents in a 4.0 mL glass vial using a magnetic stirrer. The solid in the slurry was characterized by XRPD after 5 - 7 days. 3. Solid Vapor Diffusion Approximately 30 mg of the API was held in a 4.0 mL glass vial. The vial was placed in a 20 mL glass vial containing the solvent. The solid was characterized by XRPD after 7 - 10 days. 4. Poor Solvent Addition Approximately 30 mg of the API was dissolved in a solvent to prepare a saturated solution, and a poor solvent was added up to a 10 - volume ratio. The resulting solid was characterized by XRPD. 5. Liquid Vapor Diffusion Approximately 30 mg of the API was dissolved in a solvent to prepare a saturated solution in a 4.0 mL glass vial, and this vial was placed in a 20 mL glass vial containing the poor solvent. The resulting solid was characterized by XRPD after 7 - 10 days. 6. Polymer - Induced Crystallization Approximately 30 mg of the API and 2.0 mg of the listed polymer were added to a solvent and stirring was continued at room temperature. The solid precipitated from the slurry or the solution was characterized by XRPD after 7 - 10 days. 7. Grinding: Grind approximately 30 mg of the API with a mortar and pestle over several minutes. Then, analyze the ground material by XPRD. 8. Compression: Approximately 100 mg of the API was compressed at 20 kN using a STYL , One Nano and a Type B tool. Then, crush the tablet and analyze the powdered material by XPRD.

[0243]

Table 12

[0244]

Table 13

[0245]

Table 14

[0246]

Table 15

[0247]

Table 16

[0248]

Table 17

[0249]

Table 18

[0250]

Table 19

[0251] The polymorphic form FB-4 was discovered from two test conditions, namely slurry preparation and solid vapor diffusion, using acetone as the solvent system. By DSC, two endothermic events were shown at 120.8 °C and 206.7 °C (Tpeak), and by TGA, a 4.4 wt% decrease was observed between room temperature and 175 °C. The thermal analysis data suggest that polymorphic form FB-4 is likely the hydrated form of the free base.

[0252] The polymorphic form FB-5 was discovered from the DMSO / toluene solvent system. Since the amount of material obtained from the test conditions was limited, only DSC analysis was performed, which showed two endothermic events at 132.0 °C and 221.1 °C (Tpeak), suggesting that the solvated or hydrated form of the free base is most likely.

[0253] To investigate the possibility of a solid-to-solid phase transition, morph FB-1 was tested under grinding conditions. However, XRPD data indicated no phase transition and suggested that the material is stable under grinding conditions. Morph FB-1 was also tested using aqueous and organic solvent systems to investigate its potential to form a hydrated form. Analysis of the final solid by XRPD suggested that morph FB-1 is stable under these test conditions.

[0254] The breakdown of the hydrochloride salt of compound 1 resulted in a new polymorph called morph FB-3. However, the final solids from multiple batches of salt breakdown showed some variation at a lower angle (2θ) in the XRPD diffractogram. Analysis of the solids by DSC-TGA suggested that the variation in the XRPD may be due to differences in solvate content. XRPD analysis of the ground and compressed morph FB-1 material did not show any solid-to-solid morphological changes in the data.

[0255] Equilibrium solubility test of morphology FB-1 The experimental procedure and results are summarized in Table 20. The buffer medium was prepared according to USP 35-NF 30 (page 5774), and the biorelevant medium was prepared according to the provided instructions (biorelevant.com). Approximately 40.0 mg of solid was added to 2.0 mL of medium to obtain a slurry. The system was set up at 37°C for approximately 18 hours. The solubility of the solution was tested by HPLC, and the pH was determined. A calibration curve was created using compound 1 at concentrations of 0.05–0.51 mg / mL. The regression value was found to be greater than 0.999. The calibration curve was further used to determine the solubility concentration.

[0256] [Table 20]

[0257] [Table 21]

[0258] Solid state stability of compound 1, form FB-1 The material used in this test was compound 1, form FB-1. Based on solid-state analysis and HPLC analysis, compound 1, form FB-1, is stable for 4 weeks at 25°C / 60%RH, 40°C / 75%RH, and 60°C. The experimental results are summarized in Table 22.

[0259] [Table 22]

[0260] Example 2: Salt Screening Solubility test The approximate solubility of crystalline compound 1, form FB-1, was estimated in 20 solvents, and the situation / results are reported in Table 23. procedure: 1) Weigh approximately 5 mg of the starting material and place it in a 4.0 mL vial. Add 25 μL of solvent at room temperature. 2) Shake the solution to stir it. If a clear solution is not obtained, add another 25 μL and repeat until 0.75 mL of solvent has been added. 3) Subsequently, the solvent was added in increments of 0.25 mL until the total volume reached 3.0 mL. 4) Stir the solution overnight.

[0261] [Table 23]

[0262] Salt screening of compound 1 Salt screening of crystalline compound 1 was completed at room temperature using eight acids and five solvents. The experimental procedure is described below. The experimental results are summarized in Table 24. For detailed solid-state characteristic analysis data, please refer to Appendix II.

[0263] FB+acidic CI Salt screening was set up using eight counterions in five solvent systems. Approximately 25 mg of free base was weighed into a 2.0 mL vial. Counterions (Cl) were added to the solid in a 1:1 (API:Cl) molar ratio, and finally approximately 0.25 mL of solvent was added. The sample vial was kept at room temperature while stirring. The sample was stirred at room temperature for 3 to 5 days, and any changes in the solid state were analyzed by PXRD.

[0264] [Table 24]

[0265] [Table 25]

[0266] Salt scale-up and feature analysis Hippuric-2 The scale-up of hippurate from crystalline compound 1 was tested on a sub-mg scale. The experimental results of salt scale-up are summarized in Tables 26 and 27. The results showed good reproducibility. Analysis of hippurate-2 by DVS showed a 0.74 wt% H2O increase between 0 and 80% RH and a 3.8 wt% H2O increase between 0 and 90% RH. However, the XRPD of the solid after DVS changed, which was referred to as a mixture of hippurate-2 and hippurate-4.

[0267] [Table 26]

[0268] [Table 27]

[0269] Maleic-1 The scale-up of the maleate of crystalline compound 1 was tested on a sub-mg scale. The results of the salt scale-up are summarized in Tables 28 and 29. The results show good reproducibility in the formation of maleic acid-1 salt. Maleic acid-1 showed a slightly hygroscopic increase of 1.03 wt% H2O between 0 and 90% RH. The solid after DVS remained maleic acid-1.

[0270] [Table 28]

[0271] [Table 29]

[0272] Lactic acid-1 and lactic acid-2 A test was conducted using 25 mg of FB-1 to reproduce lactate-1. The results and status of the salt scale-up are summarized below. The test successfully reproduced lactate-1.

[0273] [Table 30]

[0274] [Table 31]

[0275] The scale-up of the lactate of crystalline compound 1 was completed at room temperature on a 550 mg scale. The results and status of the salt scale-up are summarized in Tables 32 and 33. The reproducibility of the crystallization of lactate-1 was not good on a 500 mg scale, and lactate-2 was obtained at the end of the test instead of lactate-1 form.

[0276] [Table 32]

[0277] [Table 33]

[0278] Lactate-2 salt was analyzed by DVS. The data showed a 2.98 wt% increase between 0–80% RH, classifying it as a moderately hygroscopic material. Between 0–90% RH, the material showed a 3.66 wt% increase, and the XRPD of the solid after DVS was consistent with lactic acid-2.

[0279] Slurry preparation in H2O Free bases and selected 3-lead salts were slurryed in water and stirred continuously at room temperature for 5 days. After 5 days of stirring, the final solid was analyzed by XRPD. In summary, hippurates, maleates, and lactates tended to transform into different solid forms, suggesting instability of these selected salt forms when the slurry was prepared in water.

[0280] [Table 34]

[0281] Equilibrium solubility test of FB-1, hippuric acid-2, and lactate-2 The experimental procedures and results are summarized in Tables 35, 36, and 37. Buffering media were prepared according to USP 35-NF 30 (page 5774), and biorelevant media were prepared according to the provided instructions (biorelevant.com). Approximate amounts of solid were added to 0.5–1.0 mL of media to obtain slurry. The system was set up at room temperature for 24 hours. The solid was then filtered using a centrifugal filter.

[0282] The solubility of the solution was tested by HPLC, and the pH was determined. Because the concentration of the solubility sample was too low, the injection volume was adjusted from 0.3 μL to 3 μL. The final solid was analyzed by XRPD.

[0283] [Table 35]

[0284] [Table 36]

[0285] [Table 37]

[0286] Example 3: Salt Destruction Salt decomposition was performed on a 4.2g scale. The experimental results are summarized in Table 38.

[0287] [Table 38]

[0288] Development of the HPLC method Multiple methods were tested during the salt screening, polymorphism screening, and excipient compatibility testing processes. For the solubility tests reported in this report, the concentration of compound 1 was analyzed using the following HPLC method (Table 39). The HPLC methods reported in Table 40 were used for Log P and solid state stability testing.

[0289] [Table 39]

[0290] [Table 40] X-ray powder diffraction (XRPD) Equipment: Panalytical Empyrean Parameters: X-ray tube Cu (Kα radiation), Power: 45kV × 40mA Scanning range: 2~40 2θ(°) Step size: 0.01° Scanning speed: 6.33°(2θ) / min Thermogravimetric analysis (TGA) Equipment: Discovery TGA manufactured by TA Instruments Parameters: Heating at 10°C / min from 25 to 300°C, N2 sweep at 50 mL / min. Differential Scanning Calorimetry (DSC) Equipment: Discovery DSC manufactured by TA Instruments Parameters: Heat up to 300°C at a rate of 10°C / min Polarized light microscopy (PLM) Equipment: Nikon Eclipse Ci POL Camera: Nikon DS-Fi3 Software: Nikon NIS Elements 1 1H NMR Equipment: Bruker 400 MHz Ultrashield Solvent: DMSO-d6 water activity measuring device Equipment:Aqualab TDL-2 Titrator Equipment: Mettler Toledo G20S Compact Potentiometric Titrator Dynamic Vapor Sorption (DVS) Equipment: DVS Intrinsic, manufactured by Surface Measurement Systems. Parameters: 25°C, 30-90-0-90-0%RH for 2 cycles

[0291] [Table 41]

[0292] The examples and embodiments described herein are for illustrative purposes only, and in some embodiments, various modifications or changes should be included within the scope of the disclosure and the appended claims.

Claims

1. 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile: 【Chemistry 1】 or the crystalline form of its pharmaceutically acceptable salt or solvate.

2. Free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile: 【Chemistry 2】 or the crystalline form of the pharmaceutically acceptable solvate thereof.

3. Anhydrous free base 5-((5-(4-(((1R,3S)-3-aminocyclopentyl)oxy)-2-methoxy-6-methylpyridine-3-yl)-1H-pyrazole-3-yl)amino)pyrazine-2-carbonitrile: 【Transformation 3】 The crystalline form.

4. The crystalline compound 1 has the following properties: (a) A substantially the same X-ray powder diffraction (XRPD) pattern as shown in Figure 1, (b) X-ray powder diffraction (XRPD) pattern with characteristic peaks at 11.96±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, and 26.75±0.1°2θ. (c) X-ray powder diffraction (XRPD) pattern with characteristic peaks at 13.87±0.1°2θ and 17.06±0.1°2θ. (d) Differential scanning calorimetry (DSC) thermogram with endothermic properties and a peak temperature at approximately 216.5°C (start). (e) A thermogravimetric (TGA) thermogram that is substantially the same as the one shown in Figure 2. (f) A thermogravimetric analysis (TGA) thermogram showing a mass loss of approximately 0.10% from the start of heating up to approximately 100.0°C, or (g) combinations of those The crystal form according to claim 1, which is a form of free base FB-1 characterized by having at least one of the above.

5. The crystal morphology according to any one of claims 1 to 4, wherein the crystal morphology has substantially the same X-ray powder diffraction (XRPD) pattern as that shown in Figure 1.

6. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks as shown in Table 1.

7. The crystalline morphology according to any one of claims 1 to 6, wherein compound 1 of the crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 11.96±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, and 26.75±0.1°2θ.

8. The crystalline form according to any one of claims 1 to 6, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 13.87 ± 0.1°²θ and 17.06 ± 0.1°²θ.

9. The crystalline form according to any one of claims 1 to 6, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.87 ± 0.1°²θ.

10. The crystalline form according to any one of claims 1 to 6, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 17.06 ± 0.1°²θ.

11. The crystalline form according to any one of claims 1 to 10, wherein the X-ray powder diffraction (XRPD) pattern further includes a peak at 5.27 ± 0.1° 2θ.

12. The crystalline form according to any one of claims 1 to 11, wherein the X-ray powder diffraction (XRPD) pattern further includes a peak at 13.34 ± 0.1° 2θ.

13. The crystalline form according to any one of claims 1 to 12, wherein the X-ray powder diffraction (XRPD) pattern further includes a peak at 15.66 ± 0.1° 2θ.

14. The crystalline form according to any one of claims 1 to 13, wherein the X-ray powder diffraction (XRPD) pattern further includes a peak at 22.24 ± 0.1° 2θ.

15. The crystal morphology according to any one of claims 1 to 14, wherein the X-ray powder diffraction (XRPD) pattern further includes a peak at 27.84 ± 0.1° 2θ.

16. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 5.27 ± 0.1° 2θ.

17. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 11.96 ± 0.1°²θ.

18. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.34 ± 0.1°²θ.

19. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 13.87 ± 0.1°²θ.

20. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 15.66 ± 0.1° 2θ.

21. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 17.06 ± 0.1°²θ.

22. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 18.81 ± 0.1°²θ.

23. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 19.91 ± 0.1°²θ.

24. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 21.67 ± 0.1° 2θ.

25. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 22.24 ± 0.1° 2θ.

26. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 26.75 ± 0.1° 2θ.

27. The crystalline form according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., form FB-1, has an X-ray powder diffraction (XRPD) pattern with a characteristic peak at 27.84 ± 0.1°²θ.

28. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of the crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with characteristic peaks at 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 15.66±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

29. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least two characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

30. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least three characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

31. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least four characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

32. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least five characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

33. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least six characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

34. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least seven characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

35. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least eight characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

36. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least nine characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

37. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least 10 characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

38. The crystalline morphology according to any one of claims 1 to 5, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has an X-ray powder diffraction (XRPD) pattern with at least 11 characteristic peaks selected from 5.27±0.1°2θ, 11.96±0.1°2θ, 13.34±0.1°2θ, 13.87±0.1°2θ, 15.66±0.1°2θ, 17.06±0.1°2θ, 18.81±0.1°2θ, 19.91±0.1°2θ, 21.67±0.1°2θ, 22.24±0.1°2θ, 26.75±0.1°2θ, and 27.84±0.1°2θ.

39. The crystalline morphology according to any one of claims 1 to 38, wherein compound 1 of a crystalline free base, i.e., morphology FB-1, has a differential scanning calorimetry (DSC) thermogram with endothermic properties having a peak temperature at approximately 216.5°C (start).

40. The crystalline form according to any one of claims 1 to 39, wherein compound 1 of the crystalline free base, i.e., form FB-1, has substantially the same thermogravimetric (TGA) thermogram as shown in Figure 2.

41. The crystalline form according to any one of claims 1 to 40, wherein compound 1 of the crystalline free base, i.e., form FB-1, has a thermogravimetric analysis (TGA) thermogram showing a mass loss of about 0.10% from the start of heating up to about 100.0°C.

42. The crystalline form according to any one of claims 1 to 41, wherein compound 1 of the crystalline free base, i.e., form FB-1, is physically and chemically stable.

43. The crystalline form according to any one of claims 1 to 42, wherein compound 1 of the crystalline free base, i.e., form FB-1, is chemically stable.

44. A pharmaceutical composition comprising the crystalline form described in any one of claims 1 to 43 and a pharmaceutically acceptable excipient.

45. A method for treating cancer in a subject requiring treatment, comprising administering the crystalline form described in any one of claims 1 to 43 to the subject.

46. The method according to claim 45, wherein the cancer includes a solid tumor.

47. The method according to claim 45, wherein the cancer includes a locally advanced or metastatic, unresectable solid tumor.

48. The method according to any one of claims 45 to 47, wherein the cancer comprises a tumor or tumor cells having oncogene amplification.

49. The method according to claim 48, wherein the oncogene amplification includes amplification of ABL, AKT1, AKT2, ALK, androgen receptor, BRAF, CCND1, CCND2, CCND3, CCNE1, CDK12, CDK4, CDK6, EGFR, ERBB2, EZH2, FGFR1, FGFR2, FGFR3, FLT3, IDH1 / 2, JAK2, JAK3, KIT, KRAS, MDM2, MDM4, MET, MYC, MYCL, MYCN, NRAS, PDGFRA, TERT, VEGFRA, or any combination thereof.

50. The method according to claim 48 or claim 49, wherein the oncogene amplification is present on ecDNA.

51. The method according to claim 48 or claim 49, wherein the oncogene amplification is located on one or more chromosomal loci.

52. The method according to claim 48 or claim 49, wherein the oncogene amplification is derived from ecDNA.

53. The method according to any one of claims 45 to 52, wherein the cancer is ovarian cancer.

54. The method according to claim 53, wherein the ovarian cancer is platinum-resistant high-grade serous ovarian cancer, primary peritoneal cancer, or fallopian tube cancer.

55. The method according to any one of claims 45 to 52, wherein the cancer is uterine cancer.

56. The method according to claim 55, wherein the uterine cancer is high-grade endometrial cancer, serous uterine carcinoma, or uterine carcinosarcoma.

57. The method according to any one of claims 45 to 52, wherein the cancer is colorectal cancer, esophageal cancer, gastric cancer, gastroesophageal junction (GEJ) cancer, head and neck squamous cell carcinoma, liposarcoma, non-small cell lung cancer, or subtype squamous cell carcinoma.

58. The method according to any one of claims 45 to 52, wherein the cancer is neuroblastoma.

59. The method according to any one of claims 45 to 52, wherein the cancer is breast cancer, bile duct cancer, esophageal cancer, cervical squamous cell carcinoma, non-small cell lung cancer, gastric cancer, or subtype squamous cell carcinoma.

60. The method according to any one of claims 45 to 52, wherein the cancer is esophageal cancer, non-small cell lung cancer, sarcoma, or gastric cancer.

61. The method according to any one of claims 45 to 60, wherein the procedure further comprises administering an additional therapeutic agent.

62. The method according to claim 61, wherein the oncogene amplification includes CDK4, CDK6, EGFR, FGFR1, FGFR2, or FGFR3.

63. The method according to claim 62, further comprising administering a CDK4 / 6 inhibitor, an EGFR inhibitor, or an FGFR inhibitor as part of the treatment.

64. The method according to claim 63, wherein the EGFR inhibitor is erlotinib.

65. The method according to claim 63, wherein the FGFR inhibitor is pemigatinib.

66. The method according to claim 63, wherein the FGFR inhibitor is futivacinib.

67. The method according to claim 63, wherein the CDK4 / 6 inhibitor is abemaciclib.

Images