Crystalline form of a RAS inhibitor, a composition containing the same, and a method of using the same.

Abstract

This disclosure features crystalline forms of Ras inhibitors, their pharmaceutical compositions, and their use in the treatment of cancer.

Description

Background Art

[0001] The overwhelming majority of small molecule drugs act by binding to functionally important pockets on target proteins, thereby controlling the activity of the protein. For example, cholesterol-lowering agents known as statins bind to the enzyme active site of HMG-CoA reductase, thereby preventing the enzyme from engaging its substrate. The fact that many such drug / target interaction pairs are known has misled some people into believing that, given a reasonable amount of time, effort, and resources, small molecule modulators can be discovered for most, if not all, proteins. This is far from the truth. Current estimates suggest that only about 10% of all human proteins are targetable by small molecules. The other 90% are currently considered refractory or intractable to the above-described small molecule drug discovery. Such targets are generally referred to as "undruggable." These undruggable targets include a vast and largely untapped reservoir of pharmaceutically important human proteins. Therefore, there is much interest in discovering new molecular modalities that can control the function of such undruggable targets.

[0002] Ras proteins (K-Ras, H-Ras, and N-Ras) play an essential role in various human cancers and, therefore, have been well established in the literature as suitable targets for anticancer therapy. In fact, mutations in Ras proteins account for approximately 30% of all human cancers in the United States, and many of these are lethal. Dysregulation of Ras proteins due to mutant activation, overexpression, or upstream activation is common in human tumors, and mutant activation of Ras is frequently found in human cancers. For example, mutant activation of codon 12 in the Ras protein inhibits both the GAP-dependent and intrinsic hydrolysis rates of GTP, significantly skewing the population of Ras mutant proteins into the "on" (GTP-bound) state (Ras(ON)), and functions by leading to oncogenic MAPK signaling. In particular, Ras exhibits picomolar affinity for GTP and allows Ras to be activated even in the presence of low concentrations of this nucleotide. Mutations at codon 13 of Ras (e.g., G13C) and at codon 61 (e.g., Q61K) also confer oncogenic activity in some cancers.

[0003] In the past few decades, despite extensive drug discovery efforts against Ras, only two drugs targeting Ras have been approved in the United States, both of which target K-Ras G12C sotorasib and adagrasib. Further efforts are needed to identify additional pharmaceuticals for cancers driven by various Ras mutations. SUMMARY OF THE INVENTION

[0004] The present invention features crystalline forms of compounds useful in the treatment of diseases or medical conditions (e.g., cancer, Ras protein-related disorders). In one aspect, the present disclosure describes a crystalline form of Compound A:

[0005]

Chemical formula

[0006] In some embodiments, the crystalline form of compound A or its solvate is selected from Form 1, Form 2, Form 3, Form 4, Form 5, Form 6, Form 7, or Form 8. In some embodiments, the crystalline form of compound A or its solvate is Form 1. In some embodiments, the crystalline form of compound A or its solvate is Form 5.

[0007] In one embodiment, the present invention is characterized by a crystalline hydrate of compound A:

[0008] [ka]

[0009] In some embodiments, crystalline form 1 of compound A, or its solvate, has at least one peak at diffraction angles 2θ(°) at 5.2±0.5, 8.4±0.5, or 10.0±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) at 5.2±0.5, 8.4±0.5, and 10.0±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) at 10.7±0.5, 14.4±0.5, and 16.7±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) of 5.2±0.5, 8.4±0.5, 10.0±0.5, 10.7±0.5, 14.4±0.5, and 16.7±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) of 17.0±0.5, 18.4±0.5, and 20.5±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) at 5.2±0.5, 8.4±0.5, 10.0±0.5, 10.7±0.5, 14.4±0.5, 16.7±0.5, 17.0±0.5, 18.4±0.5, and 20.5±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, crystalline form 1 of compound A, or its solvate, has the X-ray powder diffraction pattern shown in Figure 1.

[0010] In some embodiments, crystalline form 1 of compound A is a hydrated solvate. In some embodiments, crystalline form 1 of compound A is a trihydrate. In some embodiments, crystalline form 1 of compound A is a mixed solvate of methanol and water.

[0011] In some embodiments, the crystalline form 1 of compound A is a mixed solvate of methanol and water, and furthermore, the unit cell has parameters a=10.8182A, b=13.3787A, c=17.6881A, α=88.4427°, β=79.5367°, γ=82.0377°, and V=2493.21A 3 It is characterized by having the following parameters.

[0012] In some embodiments, crystalline form 1 of compound A, or its solvate, exhibits endothermic onset at 66.8°C ± 0.5 and 95.8°C ± 0.5 in differential scanning calorimetry (DSC) profiles. In some embodiments, crystalline form 1 of compound A, or its solvate, has the DSC thermogram shown in Figure 3. In some embodiments, crystalline form 1 of compound A, or its solvate, exhibits a weight loss of 6.0% ± 0.5 (w / w) between ambient temperature and 100.0°C ± 0.5 in thermogravimetric analysis (TGA) profiles. In some embodiments, crystalline form 1 of compound A, or its solvate, has the TGA graph shown in Figure 4.

[0013] In some embodiments, the present invention is characterized by a crystalline form 5 of compound A, or its solvate, having at least one peak at 14.3±0.5, 16.8±0.5, or 20.5±0.5 at a diffraction angle of 2θ(°) when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, the crystalline form 5 of compound A, or its solvate, has peaks at 14.3±0.5, 16.8±0.5, and 20.5±0.5 at a diffraction angle of 2θ(°) when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, the crystalline form 5 of compound A, or its solvate, has peaks at diffraction angles 2θ(°) of 5.2±0.5, 17.0±0.5, and 20.3±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or when calculated from X-ray diffraction. In some embodiments, the crystalline form 5 of compound A, or its solvate, has peaks at 16.3±0.5, 17.1±0.5, and 18.2±0.5 at diffraction angles 2θ(°) when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, the crystalline form 5 of compound A, or its solvate, has peaks at 5.2±0.5, 14.3±0.5, 16.3±0.5, 16.8±0.5, 17.0±0.5, 17.1±0.5, 18.2±0.5, 20.3±0.5, and 20.5±0.5 at diffraction angles 2θ(°) when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, the crystalline form 5 of compound A, or its solvate, has the X-ray powder diffraction pattern shown in Figure 18. In some embodiments, the solvate is a hydrate.

[0014] In some embodiments, crystalline form 5 of compound A, or its solvate, exhibits endothermic onset at 32.1°C ± 0.5 and 81.2°C ± 0.5 in differential scanning calorimetry (DSC) profiles. In some embodiments, crystalline form 5 of compound A, or its solvate, has the DSC thermogram shown in Figure 19. In some embodiments, crystalline form 5 of compound A, or its solvate, exhibits a weight loss of 4.8% ± 0.5 (w / w) between ambient temperature and 150.0°C ± 0.5 in thermogravimetric analysis (TGA) profiles. In some embodiments, crystalline form 5 of compound A, or its solvate, has the TGA graph shown in Figure 20.

[0015] In one aspect, the present invention is characterized by a crystalline form of the tosylate of compound A, or a solvate thereof:

[0016] [ka]

[0017] In some embodiments, the crystalline form, when measured by X-ray diffraction after irradiation with Cu Kα X-rays or calculated from X-ray diffraction, has at least one peak at diffraction angles 2θ(°) of 5.9±0.5, 9.4±0.5, or 9.8±0.5. In some embodiments, the crystalline salt form of compound A, or its solvate, when measured by X-ray diffraction after irradiation with Cu Kα X-rays or calculated from X-ray diffraction, has peaks at diffraction angles 2θ(°) of 5.9±0.5, 9.4±0.5, and 9.8±0.5. In some embodiments, the crystalline salt form of compound A, or its solvate, when measured by X-ray diffraction after irradiation with Cu Kα X-rays or calculated from X-ray diffraction, has peaks at diffraction angles 2θ(°) of 10.2±0.5, 10.6±0.5, 11.0±0.5, and 16.1±0.5. In some embodiments, the crystalline salt form of compound A, or its solvate, when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction, has peaks at diffraction angles 2θ(°) of 5.9±0.5, 9.4±0.5, 9.8±0.5, 10.2±0.5, 10.6±0.5, 11.0±0.5, and 16.1±0.5. In some embodiments, the crystalline salt form of compound A, or its solvate, when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction, has peaks at diffraction angles 2θ(°) of 12.6±0.5, 12.8±0.5, 14.2±0.5, and 17.2±0.5. In some embodiments, the crystalline salt form of compound A, or its solvate, has peaks at diffraction angles 2θ(°) at 5.9±0.5, 9.4±0.5, 9.8±0.5, 10.2±0.5, 10.6±0.5, 11.0±0.5, 12.6±0.5, 12.8±0.5, 14.2±0.5, 16.1±0.5, and 17.2±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. In some embodiments, the crystalline salt form of compound A, or its solvate, has the X-ray powder diffraction pattern shown in Figure 6. In some embodiments, the crystalline salt form of compound A is a hydrate.In some embodiments, the crystalline salt form of compound A is a hemihydrate. In some embodiments, the crystalline salt form of compound A, or its solvate, exhibits endothermic onset at 99.5°C±0.5, 230.7°C±0.5, and 262.4°C±0.5 in differential scanning calorimetry (DSC) profiles.

[0018] In some embodiments, the crystalline salt form of compound A, or its solvate, has a DSC thermogram as shown in Figure 7. In some embodiments, the crystalline salt form of compound A, or its solvate, shows a weight loss of 2.7% ± 0.5 (w / w) between ambient temperature and 50.0°C ± 0.5°C, or a weight loss of 3.2% ± 0.5 (w / w) between ambient temperature and 200.0°C ± 0.5°C, in the thermogravimetric analysis (TGA) profile. In some embodiments, the crystalline salt form of compound A, or its solvate, has a TGA graph as shown in Figure 8.

[0019] In some embodiments, the pharmaceutical composition comprises a crystalline form of compound A, or a solvate thereof, and a pharmaceutically acceptable carrier or excipient. In one embodiment, the present invention relates to a crystalline form of compound A,

[0020] [ka]

[0021] The present invention relates to a method for producing a solvate thereof, the method comprising preparing a slurry of compound A in a suitable solvent or buffer to precipitate the crystalline solid of form 1, isolating the crystalline solid of form 1 from the slurry, and drying the crystalline solid of form 1. In some embodiments, the suitable solvent is a mixture of methanol and water. In some embodiments, the suitable buffer is acetate buffer (pH 4).

[0022] In one embodiment, the present invention relates to the crystalline form of compound A,

[0023] [ka]

[0024] The present invention is characterized by a method for producing a solvate thereof, the method comprising preparing a slurry of crystalline form 1 of compound A in a suitable solvent to precipitate the crystalline solid of form 5, isolating the crystalline solid of form 5 from the slurry, and drying the crystalline solid of form 5. In some embodiments, the suitable solvent is selected from the group consisting of isopropyl alcohol / water mixtures and acetone / water mixtures.

[0025] In one embodiment, the present invention relates to the crystalline form of compound A,

[0026] [ka]

[0027] The present invention relates to a method for producing a solvate thereof, the method comprising preparing a mixture of crystalline form 1 of compound A in isopropyl alcohol, precipitating a crystalline solid of form 5 by adding water, isolating the crystalline solid of form 5 from the slurry, and drying the crystalline solid of form 5.

[0028] In one aspect, the present invention relates to the crystalline form of the tosylate of compound A,

[0029] [ka]

[0030] The method is characterized by producing a solvate thereof, comprising: preparing a mixture of compound A and 4-toluenesulfonic acid in a suitable solvent or buffer to precipitate a crystalline solid of compound A tosylate; isolating the crystalline solid of compound A tosylate from the mixture; and drying the crystalline solid of compound A tosylate. In some embodiments, the method comprises heating the mixture before isolating the solid from the slurry. In some embodiments, suitable solvents are acetonitrile, water, or a mixture of acetonitrile and water.

[0031] In some embodiments, the present invention features a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of compound A of the present invention in crystalline form, or a solvate thereof, or a pharmaceutical composition. In some embodiments, the cancer comprises a Ras mutation. In some embodiments, the Ras mutation is located at position 12, position 13, or position 61, or a combination thereof. In some embodiments, the Ras protein mutation is located at a position selected from the group consisting of G12C, G12D, G12V, G12R, G13C, G13D, and Q61K, or a combination thereof. In some embodiments, the Ras mutation is located at a position selected from the group consisting of G12D, G12V, and G12R, or a combination thereof. In some embodiments, the Ras mutation is located at a position selected from the group consisting of G12D and G12V, or a combination thereof. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the present invention features a method for treating a subject requiring treatment for a Ras protein-related disorder, the method comprising administering to the subject a therapeutically effective amount of compound A in crystalline form, or a solvate thereof, or a pharmaceutical composition thereof.

[0032] In some embodiments, the present invention is characterized by a method for inhibiting intracellular Ras proteins, the method comprising contacting cells with an effective amount of compound A in crystalline form, a cultured hydrate thereof, or a pharmaceutical composition. In some embodiments, multiple Ras proteins are inhibited intracellularly. In some embodiments, the cells are cancer cells. In some embodiments, the cancer cells are pancreatic cancer cells. In some embodiments, the cancer cells are lung cancer cells. In some embodiments, the cancer cells are non-small cell lung cancer cells. In some embodiments, the cancer cells are colorectal cancer cells. In some embodiments, the Ras proteins include KRAS.

[0033] In some embodiments, the method further comprises administering additional anticancer therapy. In some embodiments, the additional anticancer therapy is an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, a mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4 / 6 inhibitor, a HER2 inhibitor, or a combination thereof. In some embodiments, the second Ras inhibitor is KRAS G12C It is an (OFF) inhibitor.

[0034] Any limitations discussed in relation to one embodiment of the present invention can be specifically conceived to apply to any other embodiment of the present invention. Furthermore, any compound or composition of the present invention can be used in any manner of the present invention, and any compound or composition of the present invention can be produced or utilized using any manner of the present invention.

[0035] Definitions and Chemical Terms In this application, unless otherwise clearly indicated by the context, (i) the term “one (a)” means “one or more”; (ii) the term “or” is used to mean “and / or” unless it is explicitly indicated that it means only alternative expressions or that such alternative expressions are mutually exclusive, however this disclosure supports the definitions that refer only to alternative expressions and to “and / or”; (iii) the terms “comprising” and “including” are understood to encompass itemized components or processes, whether presented by themselves or together with one or more additional components or processes; and (iv) where a scope is indicated, it includes endpoints.

[0036] As used herein, the term “approximately” is used to indicate that a value includes the standard deviation of the error of the device or method used to determine the value. In certain embodiments, unless otherwise specified or otherwise evident from the context (for example, if such a number may exceed 100% of the possible values), the term “approximately” refers to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1% or less in either direction (above or below) the stated value.

[0037] As used herein, the term “adjacent” in the context of describing adjacent atoms refers to divalent atoms directly bonded by a covalent bond. As used herein, “crystalline form of compound” and similar terms refer to the Ras inhibitors described herein, including the crystalline forms of the compound of formula I, its solvates, hydrates, and tautomers, whether or not they are expressly indicated.

[0038] The term "wild-type" refers to an entity with a structure or activity that is found in a "normal" state or context in nature (as opposed to mutants, diseases, modified organisms, etc.). Those skilled in the art will understand that wild-type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0039] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, including enantiomers and diastereomers, are intended. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active forms or in racemic forms. Methods for preparing optically active forms from optically active starting materials are known in the art, for example, by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers, such as olefins and C=N double bonds, may also exist among the compounds described herein, and all such stable isomers are intended in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described and may be isolated as mixtures of isomers or as separated isomers.

[0040] In some embodiments, one or more compounds described herein may exist in different tautomerized forms. As will be apparent from the context, unless explicitly excluded, references to such compounds encompass all such tautomerized forms. In some embodiments, the tautomerized form arises from the exchange of a single bond with an adjacent double bond and the accompanying transfer of protons. In certain embodiments, the tautomerized form may be a prototropic tautomer, which is a protonated state of an isomer having the same empirical formula and total charge as the reference form. Examples of moieties having a prototropic tautomerized form include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, amide-imoid acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in a heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H-, and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. In some embodiments, the tautomer morphs can be sterically fixed to one morph either in equilibrium or by appropriate substitution. In certain embodiments, the tautomer morphs arise from acetal interconversion.

[0041] Details of one or more embodiments of the present invention are described below. Other characteristics, purposes, and advantages of the present invention will become apparent from the description and claims. [Brief explanation of the drawing]

[0042] [Figure 1] A is an exemplary X-ray powder diffraction pattern of crystalline form 1 of free base compound A as a mixed solvate of methanol and water. B is an exemplary X-ray powder diffraction pattern of crystalline form 1 of free base compound A as a trihydrate solvate. [Figure 2] This is an exemplary X-ray crystal structure (asymmetrical unit shown) of crystalline form 1 of compound A free base. Hydrogen bonds are indicated by black dotted lines. For clarity, hydrogen atoms have been omitted. [Figure 3]This is an exemplary differential scanning calorimetry (DSC) thermogram of crystalline form 1 of compound A free base. [Figure 4] This is an exemplary thermogravimetric analysis (TGA) of crystalline form 1 of the free base of compound A. [Figure 5] This is an exemplary 1H NMR spectrum of crystalline form 1 of compound A free base. [Figure 6] This is an exemplary X-ray powder diffraction pattern of crystalline compound A tosylate. [Figure 7] This is an exemplary DSC thermogram of crystalline compound A tosylate. [Figure 8] This is an exemplary TGA of crystalline compound A tosylate. [Figure 9] This is an exemplary 1H NMR spectrum of the crystalline compound A tosylate. [Figure 10] This is dynamic vapor adsorption (DVS) of crystalline compound A tosylate. [Figure 11] This is an exemplary X-ray powder diffraction pattern of crystalline form 2 of compound A. [Figure 12] This is an exemplary DSC thermogram of crystalline form 2 of compound A. [Figure 13] This is an exemplary TGA of crystalline form 2 of compound A. [Figure 14] This is an exemplary X-ray powder diffraction pattern of crystalline form 3 of compound A. [Figure 15] This is an exemplary X-ray powder diffraction pattern of crystalline form 4 of compound A. [Figure 16] This is an exemplary DSC thermogram of crystalline form 4 of compound A. [Figure 17] This is an exemplary TGA of crystalline form 4 of compound A. [Figure 18] This is an exemplary X-ray powder diffraction pattern of crystalline form 5 of compound A. [Figure 19] This is an exemplary DSC thermogram of crystalline form 5 of compound A. [Figure 20] This is an exemplary TGA of crystalline form 5 of compound A. [Figure 21]This is an exemplary X-ray powder diffraction pattern of crystalline form 6 of compound A. [Figure 22] This is an exemplary DSC thermogram of crystalline form 6 of compound A. [Figure 23] This is an exemplary TGA of crystalline form 6 of compound A. [Figure 24] This is an exemplary X-ray powder diffraction pattern of crystalline form 7 of compound A. [Figure 25] This is an exemplary X-ray powder diffraction pattern of crystalline form 8 of compound A. [Figure 26] This is an overlay of exemplary X-ray powder diffraction patterns of crystalline forms 1-8 of compound A. [Modes for carrying out the invention]

[0043] compound In general, the present invention provides a crystalline form of formula I. The compound of formula I (hereinafter referred to as compound A) has the following structure:

[0044] [ka]

[0045] The crystalline form of compound A may be, for example, a free base or a tosylate. As described in the examples, crystalline form 1 of compound A, or its solvate, may have one or more peaks at diffraction angles 2θ(°) at 5.2±0.5, 8.4±0.5, 10.0±0.5, 10.7±0.5, 14.4±0.5, 16.7±0.5, 17.0±0.5, 18.4±0.5, and 20.5±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays or calculated from X-ray diffraction. Form 1 or its solvate may have the X-ray powder diffraction diagrams shown in Figures 1A and 1B. Form 1 may have the crystalline structure shown in Figure 2. The crystalline structure of Form 1 is the free base of compound A as a mixed solvate of methanol and water. Form 1, or its solvate, may exhibit endothermic onset at 66.8°C±0.5 and 95.8°C±0.5 as determined by differential scanning calorimetry (see Figure 3). Form 1, or its solvate, may exhibit a weight loss of 6.0%±0.5 (w / w) between ambient temperature and 100.0°C±0.5 in the thermogravimetric analysis profile (see Figure 4).

[0046] Crystalline form 5 of compound A, or its solvate, may have one or more peaks at diffraction angles 2θ(°) at 5.2±0.5, 14.3±0.5, 16.3±0.5, 16.8±0.5, 17.0±0.5, 17.1±0.5, 18.2±0.5, 20.3±0.5, and 20.5±0.5 when measured by X-ray diffraction after irradiation with Cu Kα X-rays, or calculated from X-ray diffraction. Form 5, or its solvate, may have the X-ray powder diffraction pattern shown in Figure 18. Form 5, or its solvate, may exhibit endothermic onset at 32.1℃±0.5 and 81.2℃±0.5 as determined by differential scanning calorimetry (DSC) (see Figure 19). Form 5, or its solvate, may exhibit a weight loss of 4.8% ± 0.5 (w / w) between ambient temperature and 150.0°C ± 0.5°C in the thermogravimetric analysis profile (see Figure 20).

[0047] The crystalline form of the tosylate of compound A, or its solvate, may have one or more peaks at diffraction angles 2θ(°) at 5.9±0.5, 9.4±0.5, 9.8±0.5, 10.2±0.5, 10.6±0.5, 11.0±0.5, 12.6±0.5, 12.8±0.5, 14.2±0.5, 16.1±0.5, and 17.2±0.5. This form, or its solvate, may have the X-ray powder diffraction pattern shown in Figure 6. This form, or its solvate, may exhibit endothermic onset at 99.5℃±0.5, 230.7℃±0.5, and 262.4℃±0.5 by DSC (see Figure 7). This form, or its solvate, may exhibit a weight loss of 2.7% ± 0.5 (w / w) between ambient temperature and 50.0°C ± 0.5°C, and a weight loss of 3.2% ± 0.5 (w / w) between ambient temperature and 200.0°C ± 0.5°C in the thermogravimetric analysis profile (see Figure 8).

[0048] A method for treating cancer in subjects requiring cancer treatment is also provided, the method comprising administering a therapeutically effective amount of the crystalline compound of the present invention to the subject. The cancer may be, for example, pancreatic cancer, colorectal cancer, non-small cell lung cancer, acute myeloid leukemia, multiple myeloma, thyroid cancer, myelodysplastic syndrome, or squamous cell carcinoma of the lung. In some embodiments, the cancer includes Ras mutations such as G12C, G12D, G12V, G12S, G13C, G13D, and Q61K. Other Ras mutations are described herein.

[0049] A method for treating Ras protein-related disorders in subjects requiring treatment of such disorders is also provided, the method comprising administering a therapeutically effective amount of the crystalline compound of the present invention to the subject.

[0050] A method for inhibiting Ras proteins within cells is also provided, which comprises contacting cells with an effective amount of the crystalline compound of the present invention. For example, the Ras proteins are Ras G12C, Ras G12D, Ras G12V, Ras G12R, Ras G13C, Ras G13D, or Ras Q61K. Other Ras proteins are described herein. The cells may be cancer cells such as pancreatic cancer cells, colorectal cancer cells, non-small cell lung cancer cells, acute myeloid leukemia cells, multiple myeloma cells, thyroid cancer cells, myelodysplastic syndrome cells, or lung squamous cell carcinoma cells. Other types of cancer are described herein. The cells may be in vivo or in vitro.

[0051] In some embodiments, the methods or uses described herein further include administering additional anticancer therapies. In some embodiments, the additional anticancer therapy is a HER2 inhibitor, an EGFR inhibitor, a second Ras inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, a PD-L1 inhibitor, a PD-1 inhibitor, a CDK4 / 6 inhibitor, or a combination thereof. In some embodiments, the additional anticancer therapy is an SHP2 inhibitor. Other additional anticancer therapies are described herein.

[0052] Synthesis method The compounds described herein may be prepared from commercially available starting materials or synthesized using known organic, inorganic, or enzymatic processes.

[0053] The compounds of the present invention can be prepared in several ways well known to those skilled in the art of organic synthesis. Exemplary synthesis of the compounds of the present invention is disclosed in WO2022 / 060836, which is incorporated herein by reference.

[0054] Pharmaceutical composition and method of use The crystalline form of the compound related to the present invention is a Ras inhibitor, which is useful in cancer treatment. Accordingly, one embodiment of the present invention provides a pharmaceutical composition comprising the crystalline form of the present invention and a pharmaceutically acceptable excipient, as well as a method for preparing such a composition using the compound of the present invention.

[0055] As used herein, the term “pharmaceutical composition” refers to a compound or crystalline form, such as the crystalline compound of the present invention, formulated with pharmaceutically acceptable excipients. In some embodiments, the crystalline form (plural) of the compound is present in the pharmaceutical composition in a unit dose appropriate for administration in a therapeutic regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to the relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, and is suitable for: oral administration, e.g., oral tablets (aqueous or nonaqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for intracellular absorption, boluses, powders, granules, and pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as sterile solutions or suspensions, or as sustained-release formulations; topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; vaginal or rectal administration, e.g., as pessaries, creams, or foams; sublingual administration; intraocular administration; transdermal administration; or suitable for nasal administration, intrapulmonary administration, and administration to other mucosal surfaces.

[0056] As used herein, “pharmaceutically acceptable excipients” refers to any inert component (e.g., a vehicle capable of suspending or dissolving an active compound) that is non-toxic and non-inflammatory in the subject matter. Typical excipients include, for example, antifouling agents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavoring agents, fragrances, lubricants (flow enhancers), preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, or hydrating water. Examples of excipients include, but are not limited to, optionally substituted butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropylcellulose, optionally substituted hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolic acid, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those skilled in the art are familiar with a wide variety of drugs and materials useful as excipients.See, for example, Ansel, et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. In some embodiments, the composition comprises at least two different pharmaceutically acceptable excipients.

[0057] As used herein, the term “subject” refers to any member of the animal kingdom. In some embodiments, “subject” refers to a human at any stage of development. In some embodiments, “subject” refers to a human patient. In some embodiments, “subject” refers to a non-human animal. In some embodiments, the non-human animal is a mammal (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, or pigs). In some embodiments, subjects may be mammals, birds, reptiles, amphibians, fish, or insects, but are not limited to these. In some embodiments, subjects may be transgenic animals, genetically modified animals, or clones.

[0058] As used herein, the term “dosage form” refers to a physically distinct unit of a compound (e.g., a crystalline compound of the present invention) for administration to a subject. Each unit contains a predetermined amount of the compound. In some embodiments, such an amount is a unit dose (or its entire fraction) appropriate for administration to a relevant population according to an administration regimen (i.e., using a therapeutic administration regimen) that has been determined to correlate with a desired or beneficial outcome. Those skilled in the art will understand that the total amount of a therapeutic composition or compound administered to a particular subject may be determined by one or more attending physicians and may involve administration of multiple dosage forms.

[0059] As used herein, the term “dosage regimen” refers to a set of unit doses (usually two or more) administered individually to a subject, usually separated by a period of time. In some embodiments, a given therapeutic compound (e.g., a crystalline compound of the present invention) has a recommended dosage regimen, which may comprise one or more doses. In some embodiments, the dosage regimen comprises multiple doses, each separated from the others by time intervals of the same length; in some embodiments, the dosage regimen comprises multiple doses, comprising at least two different time intervals separating the individual doses. In some embodiments, all doses within the dosage regimen are identical unit doses. In some embodiments, different doses within the dosage regimen are different amounts. In some embodiments, the dosage regimen comprises a first dose at a first dose, followed by one or more further doses at a second dose different from the first dose. In some embodiments, the dosage regimen comprises a first dose at a first dose, followed by one or more further doses at a second dose that is the same as the first dose. In some embodiments, the administration regimen correlates with a desired or beneficial outcome when administered across the relevant population (i.e., it is a therapeutic administration regimen).

[0060] A "treatment regimen" refers to a dosing regimen across relevant populations that correlates with a desired or beneficial therapeutic outcome. The term “treatment” (and additionally, “to treat” or “to treat”), in its broadest sense, refers to any administration of a substance (e.g., a crystalline compound of the present invention) that partially or completely alleviates, improves, reduces, inhibits, delays the onset, reduces the severity, or reduces the occurrence of one or more symptoms, characteristics, or causes of a particular disease, disorder, or condition. In some embodiments, such treatment may be administered to subjects who show no signs of the relevant disease, disorder, or condition, or to subjects who show only the initial signs of the disease, disorder, or condition. Alternatively, or in addition, in some embodiments, such treatment may be administered to subjects who show established signs of one or more of the relevant diseases, disorders, or conditions. In some embodiments, treatment may be administered to subjects diagnosed with suffering from the relevant disease, disorder, or condition. In some embodiments, treatment may be administered to subjects known to have one or more susceptibility factors that are statistically correlated with an increased risk of developing the relevant disease, disorder, or condition.

[0061] The term “therapeutic dose” means a quantity sufficient to treat a disease, disorder, or condition when administered to a population suffering from or suspected of having a disease, disorder, or condition, according to a therapeutic dosing regimen. In some embodiments, a therapeutic dose is a quantity that reduces the onset or severity of one or more symptoms of a disease, disorder, or condition, or delays the onset of one or more symptoms of a disease, disorder, or condition. Those skilled in the art will understand that the term “therapeutic dose” does not actually require that successful treatment be achieved in a particular individual. Rather, a therapeutic dose can be a quantity that, when administered to patients requiring such treatment, produces a specific desired pharmacological response in a significant number of subjects. It is specifically understood that a particular subject may actually be “refractory” to the “therapeutic dose.” In some embodiments, a reference to a therapeutic dose may refer to a quantity measured in one or more specific tissues (e.g., tissues affected by the disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). Those skilled in the art will understand that in some embodiments, the therapeutically effective dose may be formulated or administered as a single dose. In some embodiments, the therapeutically effective dose may be formulated or administered in multiple doses, for example, as part of an administration regimen.

[0062] For use as a therapeutic agent, the crystalline forms of the compounds disclosed herein can be formulated as pharmaceutical or veterinary compositions. Depending on the target being treated, the method of administration, and the type of treatment desired, e.g., prevention, prophylaxis, or therapeutic agent, the compounds are formulated in a manner consistent with these parameters. A summary of such techniques can be found in Remington: The Science and Practice of Pharmacy, 21 stThis information can be found in Edition, Lippincott Williams & Wilkins, (2005); and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JCBoylan, 1988–1999, Marcel Dekker, New York, each of which is incorporated herein by reference.

[0063] Each composition may be prepared according to conventional mixing, granulation, or coating methods, and the pharmaceutical composition may contain 0.1% to 99%, 5% to 90%, or 1% to 20% (by weight or volume) of the crystalline form of the compound of the present invention. In some embodiments, the crystalline forms of the compounds described herein may be present in total at 1 to 95% by weight of the total weight of a composition such as a pharmaceutical composition.

[0064] The composition can be provided in dosage forms suitable for intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, cutaneous, subcutaneous, topical, transdermal, sublingual, transnasal, intravaginal, intrabladderal, intraurethral, ​​intrathecal, epidural, transaural, or intraocular administration, or for injection, inhalation, or direct contact with the nasal, genitourinary, reproductive, or oral mucosa. Accordingly, the pharmaceutical composition may be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, oral medications, infiltration delivery devices, suppositories, enemas, injections, implants, sprays, preparations suitable for iontophoresis delivery, or aerosols. The composition may be formulated according to conventional pharmaceutical regulations.

[0065] As used herein, the term “administration” means the administration of a composition (e.g., a crystalline form of compound A, or a preparation containing a crystalline form of compound A as described herein) to a subject or system. Administration to an animal subject (e.g., human) may be by any suitable route. For example, in some embodiments, administration may be bronchial (including bronchial infusion), cheek, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, transnasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, intratracheal (including intratracheal infusion), transdermal, vaginal, or intravitreous.

[0066] The formulations can be prepared in a manner suitable for systemic administration or local or local administration. Systemic formulations may be designed for injection (e.g., intramuscular, intravenous, or subcutaneous injection), or they may be prepared for transdermal, transmucosal, or oral administration. The formulations generally contain diluents, and optionally adjuvants, buffers, and preservatives. The crystalline form of the compound can also be administered in liposome compositions or as microemulsions.

[0067] For injection, formulations can be prepared in conventional forms, such as a solution or suspension, or as a solid suitable for solution or suspension in liquid before injection, or as an emulsion. Suitable excipients include, for example, water, physiological saline, dextrose, and glycerol. Such compositions may also contain certain amounts of non-toxic auxiliary substances (e.g., wetting agents or emulsifiers), pH buffers, such as sodium acetate and sorbitan monolaurate.

[0068] Various sustained-release systems for drugs have also been devised. See, for example, U.S. Patent No. 5,624,677. Systemic administration may also involve relatively non-invasive methods, such as suppositories, transdermal patches, transmucosal delivery, and intranasal administration. Oral administration is also suitable for the compounds of the present invention. Preferred forms, as understood in the art, include syrups, capsules, and tablets.

[0069] The crystalline forms of each compound described herein can be formulated in various ways known in the art. For example, the first and second agents of a combination therapy can be formulated together or separately. Other modalities of combination therapy are described herein.

[0070] Individually formulated drugs can be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing two pills, pills and powder, suppositories and liquids in vials, or two topical creams. A kit may include optional components that facilitate the administration of a unit dose to a subject, such as vials for reconstituting powder forms, syringes for injection, customized intravenous delivery systems, or inhalers. In addition, a unit dose kit may include instructions for preparation or administration of the composition. A kit may be manufactured as a single-use unit dose for one subject, or as a multi-use unit dose for a specific subject (where the potency of individual compounds may change at a constant dose or as treatment progresses); or a kit may contain multi-use doses suitable for administration to multiple subjects ("bulk packaging"). The components of the kit can be assembled into cartons, blister packs, bottles, tubes, etc.

[0071] Preparations for oral use include tablets containing the active ingredient(s) in a mixture with pharmaceutically acceptable non-toxic excipients. These excipients may include, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starch containing potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulators and disintegrants (e.g., cellulose derivatives containing microcrystalline cellulose, starch containing potato starch, croscarmellose sodium, alginate, or arginine); binders (e.g., sucrose, glucose, sorbitol, acacia, arginine, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, aluminum magnesium silicate, sodium carboxymethylcellulose, methylcellulose, optionally substituted hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); as well as smoothers, lubricants, and anti-adhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients may include colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[0072] Two or more compounds can be mixed or dispensed in a tablet, capsule, or other vehicle. In one example, the first compound is contained inside the tablet, the second compound is on the outside, and a substantial portion of the second compound is released before the release of the first crystalline compound.

[0073] Formulations for oral use may be provided as chewable tablets, or as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with water or an oily medium (e.g., peanut oil, liquid paraffin, or olive oil). Powders, granules, and pellets can be prepared using the above-mentioned components below tablets and capsules by conventional methods, for example, using a mixer, fluidized bed apparatus, or spray dryer.

[0074] Dissolution or diffusion-controlled release can be achieved by appropriate coatings for tablet, capsule, pellet, or granule formation of crystalline compounds, or by incorporating crystalline compounds into an appropriate matrix. Controlled release coatings may include one or more of the above-mentioned coating materials, or shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate / butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-(optionally substituted) hydroxyl methacrylate, methacrylate hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate, or polyethylene glycol. In sustained-release matrix formulations, examples of matrix materials include hydrated methylcellulose, carnauba wax and stearyl alcohol, Carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, or halogenated fluorocarbons.

[0075] Examples of crystalline forms of the compounds of the present invention and liquid forms that can be incorporated for oral administration of compositions include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0076] Generally, when administered to humans, the oral dose of any of the crystalline compounds of the present invention depends on the properties of the crystalline compound and can be easily determined by those skilled in the art. The dose may be, for example, 0.001 mg to 2000 mg / day, 1 mg to 1000 mg / day, 5 mg to 500 mg / day, 100 mg to 1500 mg / day, 500 mg to 1500 mg / day, 500 mg to 2000 mg / day, or any range variable therefrom. In some embodiments, the dose may be 10 mg per day. In some embodiments, the dose may be 20 mg per day. In some embodiments, the dose may be 40 mg per day. In some embodiments, the dose may be 80 mg per day. In some embodiments, the dose may be 120 mg per day.

[0077] In some embodiments, the pharmaceutical composition may further contain additional compounds having antiproliferative activity. Depending on the method of administration, the compounds, or pharmaceutically acceptable salts thereof, are formulated into a suitable composition that allows for easy delivery. Each compound of the combination therapy, or a pharmaceutically acceptable salt thereof, may be formulated in various ways known in the art. For example, the first and second agents of the combination therapy may be formulated together or individually. Preferably, the first and second agents are formulated together for simultaneous or near-simultaneous administration.

[0078] It will be understood that the compounds and pharmaceutical compositions of the present invention can be formulated and used in combination therapy, that is, the compounds and pharmaceutical compositions can be formulated or administered simultaneously with, before, or after one or more other desired therapeutic agents or medical procedures. In specific combinations of treatments (therapeutic agents or procedures) using combination regimens, the suitability of the desired therapeutic agent or procedure and the desired therapeutic effect to be achieved will be taken into consideration. Furthermore, it will be understood that the treatments used may achieve the desired effect for the same disorder, or they may achieve different effects (e.g., control of any adverse effects).

[0079] As described herein, the administration of each drug in combination therapy may be independent, once to four times daily, over a period of one to one year, or even over the lifetime of the patient. Chronic, long-term administration may be indicated.

[0080] How to use In some embodiments, the present invention discloses methods for treating a disease or disorder characterized by abnormal Ras activity due to Ras variants. In some embodiments, the disease or disorder is cancer.

[0081] Accordingly, a method for treating cancer in a subject requiring treatment for cancer is also provided, the method comprising administering to the subject a therapeutically effective amount of a crystalline form of the compound of the present invention, or a pharmaceutical composition comprising such a crystalline form of the compound or a salt thereof. In some embodiments, the cancer is colorectal cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, appendiceal cancer, melanoma, acute myeloid leukemia, small intestine cancer, ampulla cancer, germ cell carcinoma, cervical cancer, cancer of unknown primary origin, endometrial cancer, esophageal cancer, GI neuroendocrine cancer, ovarian cancer, sex cord-stromal tumor cancer, hepatobiliary cancer, or bladder cancer. In some embodiments, the cancer is appendiceal cancer, endometrial cancer, or melanoma. A method for treating Ras protein-related disorders in a subject requiring treatment for Ras protein-related disorders is also provided, the method comprising administering to the subject a therapeutically effective amount of a crystalline compound of the present invention, or a pharmaceutical composition comprising such a crystalline compound or a salt thereof.

[0082] In some embodiments, the crystalline forms of the compounds of the present invention, pharmaceutical compositions containing such crystalline forms, and methods provided herein can be used to treat a wide variety of cancers, including, for example, tumors such as lung cancer, prostate cancer, breast cancer, brain cancer, skin cancer, cervical cancer, and testicular cancer. More specifically, cancers treatable by the compounds or their salts, pharmaceutical compositions containing such compounds or salts, and methods of the present invention include, but are not limited to, astrocytic cell carcinoma, breast cancer, cervical cancer, colorectal cancer, uterine cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, pharyngeal cancer, ovarian cancer, prostate cancer, and tumor types such as thyroid cancer and sarcoma. Other cancers include, for example: Cardiac malignancies, for example, non-epithelial malignancies (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyomas, fibromas, lipomas, and teratomas; Lung cancers, for example, bronchogenic carcinomas (squamous cell carcinomas, anaplastic small cell carcinomas, anaplastic large cell carcinomas, adenocarcinomas), alveolar (bronchial) carcinomas, bronchial adenomas, non-epithelial malignancies, lymphomas, chondrotoxic hamartomas, mesotheliomas; Gastrointestinal cancers, for example, esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (epithelial malignant tumor, lymphoma, leiomyosarcoma), pancreatic cancer (ductal adenocarcinoma, islet cell tumor, glucagon-producing tumor, gastrin-producing tumor, carcinoid tumor, VIP-producing tumor), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), and colorectal cancer (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma); Cancers of the urogenital organs, such as kidney cancer (adenocarcinoma, Wilms' tumor, nephroblastoma, lymphoma, leukemia), bladder and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, non-epithelial malignant tumor), and testicular cancer (seminal cell carcinoma, teratoma, embryonic carcinoma, teratocarcinoma, choriocarcinoma, non-epithelial malignant tumor, stromal cell carcinoma, fibroma, fibroadenoma, adenoid tumor, lipoma); Liver cancer, for example, hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract cancer, for example, gallbladder cancer, ampulla cancer, bile duct cancer; Bone, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulosarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteochondroma), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor; Nervous system, for example, skull (osteoma, hemangioma, granuloma, xanthomas, degenerative osteitis), meninges (meningioma, meningiosarcoma, gliomas), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal glandoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal neurofibroma, neurofibromatosis type 1, meningioma, glioma, non-epithelial malignant tumors); Gynecology, for example, uterus (endometrial cancer, uterine cancer, endometrial cancer of the uterine body), cervix (cervical cancer, pre-tumor cervical dysplasia), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified cancer), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, undifferentiated embryonal cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, sarcoma botryoides (fetal rhabdomyosarcoma)), fallopian tube (epithelial malignant tumor); Hematological, for example, blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia); myeloproliferative disorders (for example, myelofibrosis and myeloproliferative neoplasia); multiple myeloma; myelodysplastic syndrome, Hodgkin's disease, non-Hodgkin lymphoma (malignant lymphoma); Skin, for example, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and adrenal cancer, for example, neuroblastoma.

[0083] In some embodiments, the Ras protein is wild-type (Ras WT ). Thus, in some embodiments, the crystalline compound of the present invention is used in a method for treating a patient having a cancer comprising Ras WT (for example, K-Ras WT , H-Ras WT or N-Ras WT ). In some embodiments, the Ras protein is Ras amplification (for example, K-Ras amp ). Thus, in some embodiments, the crystalline compound of the present invention is used in a method for treating a patient having a cancer comprising Ras amp (K-Ras amp , H-Ras amp or N-Ras amp ). In some embodiments, the cancer comprises a Ras mutation such as a Ras mutation described herein. In some embodiments, the mutation is selected from: (a) The following K-Ras variants: G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V, and combinations thereof; (b) The following H-Ras variants: Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R, and combinations thereof; and (c) The following N-Ras variants: Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T, and combinations thereof; Or any combination of the above. In some embodiments, the cancer includes a K-Ras mutation selected from the group consisting of G12C, G12D, G13C, G12V, G13D, G12R, G12S, Q61H, Q61K, and Q61L. In some embodiments, the cancer includes an N-Ras mutation selected from the group consisting of G12C, Q61H, Q61K, Q61L, Q61P, and Q61R. In some embodiments, the cancer includes an H-Ras mutation selected from the group consisting of Q61H and Q61L. In some embodiments, the cancer includes a Ras mutation selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V. In some embodiments, the cancer contains at least two Ras mutations selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V. In some embodiments, the crystalline compound of the present invention inhibits two or more Ras mutants. For example, the compound may inhibit both K-Ras G12C and K-Ras G13C. The compound may inhibit both N-Ras G12C and K-Ras G12C. In some embodiments, the crystalline compound may inhibit both K-Ras G12C and K-Ras G12D. In some embodiments, the crystalline compound may inhibit both K-Ras G12V and K-Ras G12C. In some embodiments, the crystalline compound may inhibit both K-Ras G12V and K-Ras G12S. In some embodiments, the compound may inhibit both K-Ras G12D and K-Ras G12V. In some embodiments, the crystalline compounds of the present invention, in addition to one or more additional Ras mutations, WT Inhibits (for example, K, H, or N-Ras) WT And, K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V;K, H, or N-Ras WTH-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H, or N-Ras WT (and N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T). In some embodiments, the crystalline compounds of the present invention are Ras in addition to one or more additional Ras mutations. amp Inhibits (for example, K, H or N-Ras amp And, K-Ras G12D, G12V, G12C, G13D, G12R, G12A, Q61H, G12S, A146T, G13C, Q61L, Q61R, K117N, A146V, G12F, Q61K, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V;K, H, or N-Ras amp H-Ras Q61R, G13R, Q61K, G12S, Q61L, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, Q61H, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K, H, or N-Ras amp (and N-Ras Q61R, Q61K, G12D, Q61L, Q61H, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, Q61P, A59D, E132K, E49K, T50I, A146V, or A59T).

[0084] Methods for detecting Ras mutations are known in the art. Such methods include, but are not limited to, direct sequencing and the use of high-sensitivity diagnostic assays (CE-IVD marked). High-sensitivity diagnostic assays are described, for example, Domagala, et al., Pol J Pathol 3:145-164 (2012) (the contents of which are incorporated herein by reference in their entirety), and include TheraScreen PCR; AmoyDx; PNAClamp; RealQuality; EntroGen; LightMix; StripAssay; Hybcell plexA; Devyser; Surveyor; Cobas; and TheraScreen Pyro. See also, for example, WO2020 / 106640.

[0085] In some embodiments, the cancer is non-small cell lung cancer, and the Ras mutation includes K-Ras mutations such as K-Ras G12C, K-Ras G12V, or K-Ras G12D. In some embodiments, the cancer is colorectal cancer, and the Ras mutation includes K-Ras mutations such as K-Ras G12C, K-Ras G12V, or K-Ras G12D. In some embodiments, the cancer is pancreatic cancer, and the Ras mutation includes K-Ras mutations such as K-Ras G12D or K-Ras G12V. In some embodiments, the cancer is pancreatic cancer, and the Ras mutation includes N-Ras mutations such as N-Ras G12D. In some embodiments, the cancer is melanoma, and the Ras mutation includes N-Ras mutations such as N-Ras Q61R or N-Ras Q61K. In some embodiments, the cancer is non-small cell lung cancer, and the Ras protein is K-Ras amp In any of the above, if not already specified, the compound is also Ras WT (For example, K, H or N-Ras WT ), or Ras amp (For example, K, H or N-Ras amp ) may be inhibited.

[0086] In some embodiments, cancer is caused by Ras mutations and STK11 LOF , including KEAP1, EPHA5, or NF1 mutations. In some embodiments, the cancer is non-small cell lung cancer and includes the K-Ras G12C mutation. In some embodiments, the cancer is non-small cell lung cancer and includes the K-Ras G12C mutation and STK11 LOF The mutation is included. In some embodiments, the cancer is non-small cell lung cancer, and the K-Ras G12C mutation and STK11 LOF Includes mutations. In some embodiments, the cancer is a K-Ras G13C Ras mutation and STK11 LOF , including KEAP1, EPHA5, or NF1 mutations. In some embodiments, the cancer is non-small cell lung cancer and includes the K-Ras G12D mutation. In some embodiments, the cancer is non-small cell lung cancer and includes the K-Ras G12V mutation. In some embodiments, the cancer is colorectal cancer and includes the K-Ras G12D or G12V mutation. In some embodiments, the cancer is pancreatic cancer and includes the K-Ras G12D or K-Ras G12V mutation. In some embodiments, the cancer is pancreatic cancer and includes the K-Ras G12D mutation. In some embodiments, the cancer is pancreatic cancer and includes the K-Ras G12V mutation. In some embodiments, the cancer is endometrial cancer, ovarian cancer, cholangiocarcinoma, or appendiceal mucinous carcinoma and includes the K-Ras G12D or G12V mutation. In some embodiments, the cancer is endometrial cancer, ovarian cancer, cholangiocarcinoma, or appendiceal mucinous carcinoma and includes the K-Ras G12D mutation. In some embodiments, the cancer is endometrial cancer, ovarian cancer, cholangiocarcinoma, or appendiceal mucinous carcinoma and contains the K-Ras G12V mutation. In some embodiments, the cancer is gastric cancer and contains the K-Ras G12D or G12V mutation. In some embodiments, the cancer is gastric cancer and contains the K-Ras G12D mutation. In some embodiments, the cancer is gastric cancer and contains the K-Ras G12V mutation. In any of the foregoing, the compound is similarly Ras WT (For example, K, H or N-Ras WT ) or Ras amp (For example, K, H or N-Rasamp ) may be inhibited.

[0087] A method for inhibiting the Ras protein within a cell is also provided, the method comprising contacting a cell with an effective amount of the crystalline compound of the present invention. A method for inhibiting RAF-Ras binding is also provided, the method comprising contacting a cell with an effective amount of the crystalline compound of the present invention. The cells may be cancer cells. The cancer cells may be any type of cancer described herein. The cells may be in vivo or in vitro.

[0088] Combination therapy The methods of the present invention may include crystalline or salt forms of the compounds of the present invention used alone or in combination with one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents). When administered alone, one or more doses of the additional therapies (e.g., non-pharmacological treatments or therapeutic agents) may be reduced from the standard dose. For example, the dose may be determined empirically from the combination and order of drugs, or estimated by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[0089] The crystalline form of the compound of the present invention may be administered before, after, or concurrently with one or more additional therapies. When combined, the dosage of the crystalline compound of the present invention and the dosage of one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents) provide a therapeutic effect (e.g., a synergistic or additive therapeutic effect). The crystalline compound of the present invention and additional therapies (e.g., anticancer agents) may be administered together (e.g., in a single pharmaceutical composition) or separately, and if administered separately, they may be administered simultaneously or sequentially. Such sequential administrations may be close in time or far apart.

[0090] In some embodiments, additional therapy involves the administration of side effect limiting agents (e.g., agents intended to reduce the occurrence or severity of side effects of treatment). For example, in some embodiments, the crystalline compounds of the present invention may also be used in combination with therapeutic agents for treating nausea. Examples of agents that may be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0091] In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy). In some embodiments, one or more additional therapies include therapeutic agents (e.g., compounds or biological agents that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy) and therapeutic agents (e.g., compounds or biological agents that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In other embodiments, one or more additional therapies include two therapeutic agents. In yet another embodiment, one or more additional therapies include three therapeutic agents. In some embodiments, one or more additional therapies include four or more therapeutic agents.

[0092] In this section on combination therapies, all references to the listed medications, whether explicitly stated or not, are incorporated by reference. Non-pharmacological therapy Examples of non-pharmacological treatments include, but are not limited to, radiation therapy, cryotherapy, hyperthermia, surgery (e.g., surgical excision of tumor tissue), and T-cell adoptive transfer (ACT) therapy.

[0093] In some embodiments, the compounds of the present invention may be used as adjuvant therapy after surgery. In some embodiments, the compounds of the present invention may be used as preoperative adjuvant therapy before surgery.

[0094] Radiotherapy may be used in subjects (e.g., mammals (e.g., humans)) to inhibit abnormal cell proliferation or to treat hyperproliferative disorders such as cancer. Techniques for administering radiotherapy are known in the art. Radiotherapy can be administered via one or a combination of several methods, including, but not limited to, external beam therapy, internal radiation therapy, implanted radiation, stereotactic radiotherapy, total body radiation therapy, radiotherapy, and permanent or temporary intratissue brachytherapy. As used herein, the term “brachytherapy” refers to radiotherapy delivered by spatially restricted radioactive material inserted into the body at or near a tumor or other site of proliferative tissue disease. The term is intended to include, non-limitingly, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioisotopes of Lu). Suitable radiation sources for use as cell modifiers of the present invention include both solid and liquid forms. For non-limiting illustrative purposes, the radiation source may be a radionuclide (such as I-125, I-131, Yb-169, Ir-192 as a solid source, or I-125 as a solid source), or other radionuclides that emit photons, beta particles, gamma rays, or other therapeutic radiation. The radioactive material may also be a fluid prepared from any solution of the radionuclide(s), e.g., a solution of I-125 or I-131, or the radioactive fluid may be produced using a suitable fluid slurry containing small particles of a solid radionuclide such as Au-198 or Y-90. Furthermore, the radionuclide(s) may be embodied in a gel or radioactive microspheres.

[0095] In some embodiments, the compounds of the present invention can make abnormal cells more sensitive to radiotherapy aimed at killing or inhibiting the proliferation of such cells. Therefore, the present invention further relates to a method for increasing the sensitivity of abnormal cells in mammals to radiotherapy, the method comprising administering to a mammal an amount of the crystalline compound of the present invention effective in increasing the sensitivity of abnormal cells to radiotherapy. The amount of the compound in this method can be determined according to the means for determining an effective amount of such compound described herein. In some embodiments, the compounds of the present invention may be used as adjuvant therapy after radiotherapy or as neoadjuvant therapy before radiotherapy.

[0096] In some embodiments, the non-pharmacological treatment is T cell adoptive transfer (ACT) therapy. In some embodiments, the T cells are activated T cells. The T cells may be modified to express a chimeric antigen receptor (CAR). CAR-modified T (CAR-T) cells can be generated by any method known in the art. For example, CAR-T cells can be generated by introducing a suitable expression vector encoding a CAR into T cells. The source of T cells is obtained from a subject before proliferation and genetic modification of the T cells. T cells can be obtained from multiple sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymic tissue, tissue from infection sites, ascites, pleural fluid, spleen tissue, and tumors. In certain embodiments of the present invention, any number of T cell lines available in the art may be used. In some embodiments, the T cells are autologous T cells. Whether before or after genetic modification of T cells to express a desired protein (e.g., CAR), T cells are typically, for example, U.S. Patent Nos. 6,352,694, 6,534,055, 6,905,680, 6,692,964, 5,858,358, 6,887,466, 6,905,681, and 7 It can be activated and propagated using the methods described in Nos. 144,575, 7,067,318, 7,172,869, 7,232,566, 7,175,843, 7,572,631, 5,883,223, 6,905,874, 6,797,514, and 6,867,041.

[0097] Therapeutic drugs The therapeutic agent may be a compound used to treat cancer or related conditions. The crystalline compound of the present invention may be used in combination with a second, third, or fourth therapeutic agent, or more therapeutic agents. The crystalline compound of the present invention may be used in combination with one or more therapeutic agents, along with one or more nonpharmacological therapies.

[0098] For example, the therapeutic agent may be a steroid. Steroids are known in the art. Therefore, in some embodiments, one or more additional therapies include steroids. Preferred steroids include 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, crocortol, cloprednol, corticosterone, cortisone, cortivazole, deflazacort, desonide, desoxymethasone, dexamethasone, diflorasone, diflucortol, difprednate, enoxolone, fluazacort, fluchloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, flucortin butyl, flucortolone, fluorometholone, fluperolone acetate, flupredniden acetate, fluprednisolone, and flulandrenolide. This may include, but is not limited to, fluticasone propionate, formocortal, halcinonide, halobetazole propionate, halomethasone, hydrocortisone, loteprednol etavonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, sodium prednisolone phosphate, prednisone, prednival, prednylidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexaacetonide, and salts or derivatives thereof.

[0099] Further examples of therapeutic agents that may be used in combination therapy with the crystalline compounds of the present invention include compounds described in the following patents: U.S. Patent Nos. 6,258,812, 6,630,500, 6,515,004, 6,713,485, 5,521,184, 5,770,599, 5,747,498, 5,990,141, 6,235,764, and 8,623,885, as well as international patent applications. WO01 / 37820, WO01 / 32651, WO02 / 68406, WO02 / 66470, WO02 / 55501, WO04 / 05279, WO04 / 07481, WO04 / 07458, WO04 / 09784, WO02 / 59110, WO99 / 45009, WO00 / 59509, WO99 / 61422, WO00 / 12089, and WO00 / 02871.

[0100] The therapeutic agent may be a biological agent used in the treatment of cancer or related conditions (e.g., cytokines (e.g., interferon or interleukin, e.g., IL-2)). Biological agents are known in the art. In some embodiments, the biological agent is a biological agent of the immunoglobulin system, e.g., monoclonal antibodies (e.g., humanized antibodies, fully human antibodies, Fc fusion proteins, or functional fragments thereof) that inflict pain on a target to stimulate an anti-cancer response or antagonize antigens important to cancer. Antibody-drug conjugates are also included.

[0101] The therapeutic agent may be a T-cell checkpoint inhibitor. Such checkpoint inhibitors are known in the art. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, for example, a humanized or fully human antibody. In some embodiments, the checkpoint inhibitor is a fusion protein, e.g., an Fc receptor fusion protein. In some embodiments, the checkpoint inhibitor is a drug such as an antibody that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is a drug such as an antibody that interacts with a ligand of a checkpoint protein. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA-4 antibody or fusion protein). In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PD-L2 (e.g., PD-L2 / Ig fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligands, or combinations thereof.In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi / Regeneron), PD-L1 antibody, e.g., avelumab, durvalumab, atezolizumab, pizilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or Preusser, M. et al. The checkpoint inhibitors disclosed in al. (2015) Nat. Rev. Neurol. (including, but not limited to, ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514 / MEDI0680, BMS936559, MEDl4736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirirumab, IPH2101, 1-7F9, and KW-6002) are disclosed.

[0102] The therapeutic agent may be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB154, MTIG7192A, or OMP-313M32 (ethigirimab). Other anti-TIGIT antibodies are known in the art.

[0103] The therapeutic agent may be a drug that treats cancer or related conditions (e.g., cytotoxic agents, non-peptide small molecules, or other compounds useful for treating cancer or related conditions, collectively referred to as "anticancer agents"). Anticancer agents may be, for example, chemotherapeutic agents or targeted therapy agents. Such agents are known in the art.

[0104] Anticancer agents include mitotic inhibitors, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological reaction modifiers, alkylating agents, antimetabolites, folate analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxin, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracendione-substituted ureas, methylhydrazine derivatives, corticosteroids, progestins, estrogens, anti-estrogens, androgens, anti-androgens, and gonadotropin-releasing hormone analogs. Further anticancer agents include leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, one or more additional therapies comprise two or more anticancer agents. Two or more anticancer agents can be used in a mixture and administered in combination or separately. Suitable administration regimens for combination anticancer agents are known in the art and are described, for example, in Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999) and Douillard et al., Lancet 355 (9209):1041-1047 (2000).

[0105] Other non-exclusive examples of anticancer drugs include Gleevec® (imatinib mesylate); Kyprolis® (carfilzomib); Velcade® (bortezomib); Casodex (bicalutamide); Iressa® (gefitinib); alkylating agents, e.g., thiotepa and cyclophosphamide; alkyl sulfonates, e.g., busulfan, improsulfan, and pigosulfan; aziridines, e.g., benzodopa, carbocon, metsuredopa, and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomellamine; acetogenins (especially bratacin and bratacinone); camptothecin (including its synthetic analog topotecan); bryostatin; calistatin; CC-1065 (its adzeresin, carzeresin, and bi (Including zelesin synthetic analogs); cryptophycin (specifically cryptophycin 1 and cryptophycin 8); drastatin; duocalmycin (including synthetic analogs, KW-2189 and CB1-TM1); eryuterobin; pancratistatin; sarcodictiin A; spongistatin; nitrogen mustard, e.g., chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechlor Retamine oxide hydrochloride, melphalan, nobenbitin, fenestrine, prednimustine, trophosphamide, uracil mustard; nitrosourea, e.g., camulstine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, e.g., engine antibiotics (e.g., calicheamicin, e.g., calicheamicin gamma II and calicheamicin omega II (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); dynemicins such as dynemicin A; bisphosphonates such as clodronate; esperamicin;Neocardinostatin chromophore and related pigment protein enediin antibiotic chromophore, acrasinomycin, actinomycin, autoramycin, azaserin, bleomycin, kactinomycin, calicheamicin, carabicin, kaminomycin, carminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detrubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino - Doxorubicin, deoxydoxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin C and other mitomycins, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potophyllomycin, puromycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, pteropterin, and trimethrexate; f Purine analogs such as rudarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and phloxuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; anti-adrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid infusions such as fluphosphate; acegraton; aldofamide glycoside; aminolevulinic acid Acids; Enyluracil; Amsacrine; Bestrabusil; Bisanthren; Edatrexate; Defofamine; Demecolsin; Diadiquan; Elfomithine; Erliptinium acetate; Epotilon B and other Epotilons; Etoglucid; Gallium nitrate; Hydroxyurea; Lentinan; Mytansinoids such as Ronidynin, Mytansin and Ansamitosin; Mitoguazone; Mitoxantrone; Mopidamole; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Rosoxantrone; Podophyllic acid;2-Ethylhydrazide; Procarbazine; PSK® Polysaccharide Complex (JHS Natural Products, Eugene, OR); Lazoxane; Rhizoxin; Schizophyllan; Spirogermanium; Tenuazonic Acid; Triadicone; 2,2',2''-Trichlorotriethylamine; Trichothecenes such as T-2 toxin, Veraculine A, Loridine A, and Anguidine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Cyclophosphine Amids; thiotepas; taxoids, e.g., Taxol® (paclitaxel), Abraxane® (chromophore-free, albumin-modified nanoparticle formulation of paclitaxel), and Taxotere® (doxetaxel); chlorambucil; tamoxifen (Nolvadex®); raloxifene; aromatase-inhibiting 4(5)-imidazole; 4-hydroxytamoxifen; trioxyfen; keoxyfen; LY 117018; Onapristone; Toremifene (Fareston®); Flutamide, Niltamide, Bicalutamide, Leuprolide, Goserelin; Chlorambucil; Gemzar® Gemcitabine; 6-Thiogunine; Mercaptopurine; Platinum-coordinated complexes such as cisplatin, oxaliplatin, and carboplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitoxantrone; Vincristine; Navelbine® (Vinorelbine); Novantrone; Teniposide; Edatrexate; Daunomycin; Aminopterin; Ibandronate; Irinotecan (e.g., CPT-11); Topoisomerase inhibitor RFS Examples include difluoromethylornithine (DMFO); retinoids such as retinoic acid; esperamicin; capecitabine (e.g., Xeloda®); and any pharmaceutically acceptable salt of any of the above.

[0106] Non-exclusive examples of additional anticancer drugs include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, Avisin, avagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, and alafara. Zin, Arbocidib, 3-aminopyridine-2-carboxyaldehyde thiosemicarbazone, Amonafide, Anthracendione, Anti-CD22 immunotoxin, Antineoplastic agents (e.g., cell cycle nonspecific antineoplastic agents, and other antineoplastic agents described herein), Antitumor herbs, Apadicone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW2992, Bilicodal, Brostalicin, Briostatin, Butionine sulfoximine, CBV (chemotherapy) Kalikrin, dichloroacetic acid, discodermolide, erusamitrusine, enocitabine, eribulin, exatecan, exislind, ferginol, forodesine, phosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indocarbazole, irofluben, lanikidal, larotaxel, lenalidomide, lucanton, lulutotecan, maphosphamide, mitozolomid, napoxidine, nedaplatin, olaparib, ortata Examples include Xicel, PAC-1, Pawpaw, Pixantrone, proteasome inhibitors, rebeccamycin, reximod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swinesonin, talaporfin, talikidal, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, bajimezan, vinflunin, ZD6126, and zoskidal.

[0107] Further non-limiting examples of anticancer drugs include natural products, e.g., vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mitramycin), mitomycin, and enzymes (e.g., L-asparagine). L-asparaginase (which does not give cells that do not metabolize and synthesize asparagine of their own), antiplatelet agents, antiproliferative / antimitotic alkylating agents, e.g., nitrogen mustard (e.g., mechloretamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., CDK4 / 6 inhibitors, e.g., abemaciclib, ribociclib, palbociclib;Cericiclib (UCN-01, P1446A-05, PD-0332991, Dinacyclib (P27-00, AT-7519, RGB286638, and SCH727965)), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and its analogs, and streptozocin), trazeneth-dacarbazine (DTIC), antiproliferative / antimitotic antimetabolites (e.g., folate analogs, pyrimidine analogs (e.g., fluorouracil, phloxuridine, and cytarabine)), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine) , aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), platinum-coordinate complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apisidan, suberoylanilide hydroamic acid, vorinostat, bellinostat, LBH589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., bistusertib, temsirolimus, everolimus, ridafololimus, and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis®), PI3K inhibitors, e.g., PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), copanlisib, alpelisib and idelalisib;Multiple kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormone agonists, e.g., luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNT0328), telomerase inhibitors (e.g., GRN 163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CSl (e.g., elotuzumab), HSP90 inhibitors (e.g., 17AAG and KOS) Examples include 953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zanestra®), anti-CD138 (e.g., BT062), Torcl / 2-specific kinase inhibitors (e.g., INK128), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2 antagonists.

[0108] In some embodiments, the anticancer agent is selected from mechloretamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any analogue or derivative variant thereof.

[0109] In some embodiments, the anticancer agent is a HER2 inhibitor. HER2 inhibitors are known in the art. Non-limiting examples of HER2 inhibitors include monoclonal antibodies, e.g., trastuzumab (Herceptin®) and pertuzumab (Perjeta®); small molecule tyrosine kinase inhibitors, e.g., gefitinib (Iressa®), erlotinib (Tarceva®), pyritinib, CP-654577, CP-724714, canertinib (CI 1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788, BMS-599626, HKI-357, BIBW2992, ARRY-334543, and JNJ-26483327.

[0110] In some embodiments, the anticancer agent is an ALK inhibitor. ALK inhibitors are known in the art. Non-limiting examples of ALK inhibitors include ceritinib, TAE-684 (NVP-TAE694), PF02341066 (crizotinib or 1066), alectinib; brigatinib; entrectinib; ensartinib (X-396); lorlatinib; ASP3026; CEP-37440; 4SC-203; TL-398; PLB1003; TSR-011; CT-707; TPX-0005; and AP26113. Additional examples of ALK kinase inhibitors are described in Examples 3-39 of WO05016894.

[0111] In some embodiments, the anticancer agent is an inhibitor of a downstream member of the receptor tyrosine kinase (RTK) / growth factor receptor (e.g., SHP2 inhibitors (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SHP3809, PF-07284892, or BBP-398)), or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, These include pharmacokinetics (or tautomers), SOS1 inhibitors (e.g., BI-1701963, BI-3406, SDR5, BAY-293, MRTX-0902, or RMC-5845, or their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers), Raf inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, AKT inhibitors, or mTOR inhibitors (e.g., mTORC1 inhibitors or mTORC2 inhibitors). In some embodiments, the anticancer agent is JAB-3312.

[0112] In some embodiments, the anticancer agent is an SOS1 inhibitor. SOS1 inhibitors are known in the art. In some embodiments, the SOS1 inhibitor is WO2022219035, WO2022214594, WO2022199670, WO2022146698, WO2022081912, WO2022058344, WO2022026465, WO2022017519, WO2021173524, WO2021130731, WO2021127429, WO2021092115, WO202110596 The compounds disclosed in WO2021074227, WO2020180768, WO2020180770, WO2020173935, WO2020146470, WO2019201848, WO2019122129, WO2018172250, and WO2018115380, or selected from their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers. In some embodiments, the crystalline compounds of the present invention are used in combination with SOS1 inhibitors to treat K-Ras G13C cancer.

[0113] In some embodiments, the anticancer agent is an additional Ras inhibitor or Ras vaccine, or another therapeutic modality designed to directly or indirectly reduce the oncogenic activity of Ras. Such agents are known in the art. In some embodiments, the anticancer agent is a further Ras inhibitor. In some embodiments, the Ras inhibitor targets Ras in its active or GTP-bound state. In some embodiments, the Ras inhibitor targets Ras in its inactive or GDP-bound state. In some embodiments, the Ras inhibitor is a K-Ras inhibitor such as AMG 510, MRTX1257, MRTX849, JNJ-74699157, LY3499446, ARS-1620, ARS-853, BPI-421286, LY3537982, JDQ443, JAB-3312, JAB-21822, JAB-21000, IBI351, ERAS-3490, RMC-6291, BI 1823911, D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293, BBO-8520 (ON / OFF inhibitor), FMC-376 (ON / OFF inhibitor), GEC255, or GDC-6036. It is a G12C inhibitor. In some embodiments, the Ras inhibitor is a K-Ras G12D inhibitor such as MRTX1133, JAB-22000, MRTX282, ERAS-4, ERAS-5024, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-7835, RMC-9805, GFH375 (VS-7375), INCB161734, and KD-8. In some embodiments, the Ras inhibitor is a K-Ras G12V inhibitor such as JAB-23000. In some embodiments, the KRAS(OFF) inhibitor is a pan-RAS(OFF) inhibitor. In certain embodiments, the pan-KRAS(OFF) inhibitor is JAB-23400, JAB-23425, BI-2493, BI-2865, QTX-3034 (G12D preferred), QTX3544 (G12V preferred), ZG2001, BBO-a, BBO-B, or Pan KRas-IN-1. In some embodiments, the Ras inhibitor isIn some embodiments, the Ras inhibitor is JAB-23400. In some embodiments, the Ras inhibitor is LUNA18. In some embodiments, the Ras inhibitor is BI-2493. In some embodiments, the Ras inhibitor is a K-Ras G12V inhibitor such as JAB-23000. In some embodiments, the Ras inhibitor is selected from the Ras(ON) inhibitors (i.e., Ras in the GTP-bound state) disclosed below, which are all incorporated herein by reference, or pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers thereof: WO2022235870, WO2022235864, WO2021091982, WO2021091967, WO2021091956, and WO2020132597. Other examples of Ras inhibitors are known in the art, for example, listed below, all of which are incorporated herein by reference: WO2023287896, WO2023287730, WO2023284881, WO2023284730, WO2023284537, WO2023283933, WO2023283213, WO20232 80960, WO2023280280, WO2023280136, WO2023280026, WO2023278600, WO2023274383, WO20232743 24, WO2023034290, WO2023020523, WO2023020521, WO2023020519, WO2023020518, WO2023018812, WO2023018810, WO2023018809, WO2023018699, WO2023015559, WO2023014979, WO2023014006, WO2 023010121, WO2023009716, WO2023009572, WO2023004102, WO2023003417, WO2023001141, WO2023 001123, WO2022271923, WO2022271823, WO2022271810, WO2022271658, WO2022269508, WO2022266 167, WO2022266069, WO2022266015, WO2022265974, WO2022261154, WO2022261154, WO2022251576,WO2022251296、WO2022237815、WO2022232332、WO2022232331、WO2022232320、WO2022232318、WO2022223037、WO2022221739、WO2022221528、WO2022221386、WO2022216762、WO2022192794、WO2022192790、WO2022188729、WO2022187411、WO2022184178、WO2022173870、WO2022173678、WO2022135346、WO2022133731、WO2022133038、WO2022133345、WO2022132200、WO2022119748、WO2022109485、WO2022109487、WO2022066805、WO2022002102、WO2022002018、WO2021259331、WO2021257828、WO2021252339、WO2021248095、WO2021248090、WO2021248083、WO2021248082、WO2021248079、WO2021248055、WO2021245051、WO2021244603、WO2021239058、WO2021231526、WO2021228161、WO2021219090、WO2021219090、WO2021219072、WO2021218939、WO2021217019、WO2021216770、WO2021215545、WO2021215544、WO2021211864、WO2021190467、WO2021185233、WO2021180181、WO2021175199、2021173923、WO2021169990、WO2021169963、WO2021168193、WO2021158071、WO2021155716、WO2021152149、WO2021150613、WO2021147967、WO2021147965、WO2021143693、WO2021142252、WO2021141628、WO2021139748、WO2021139678、WO2021129824、WO2021129820、WO2021127404、WO2021126816、WO2021126799、WO2021124222、WO2021121371, WO2021121367, WO2021121330, WO2020050890, WO2020047192, WO2020035031, WO202002870 6, WO2019241157, WO2019232419, WO2019217691, WO2019217307, WO2019215203, WO2019213526, WO20192135 16, WO2019155399, WO2019150305, WO2019110751, WO2019099524, WO2019051291, WO2018218070, WO201821 7651, WO2018218071, WO2018218069, WO2018206539, WO2018143315, WO2018140600, WO2018140599, WO20181 40598, WO2018140514, WO2018140513, WO2018140512, WO2018119183, WO2018112420, WO2018068017, WO201 8064510, WO2017201161, WO2017172979, WO2017100546, WO2017087528, WO2017058807, WO2017058805, WO20 17058728, WO2017058902, WO2017058792, WO2017058768, WO2017058915, WO2017015562, WO2016168540, WO2016164675, WO2016049568, WO2016049524, WO2015054572, WO2014152588, WO2014143659 and WO2013155223.

[0114] In some embodiments, therapeutic agents that can be combined with the crystalline compounds of the present invention are inhibitors of the MAP kinase (MAPK) pathway (or "MAPK inhibitors"). Such agents are known in the art. Examples of MAPK inhibitors include, but are not limited to, one or more MAPK inhibitors described in Cancers (Basel) 2015 Sep;7(3):1758-1784. For example, MAPK inhibitors include trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimacertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA 119 / BAY 86-9766); GDC-0973 / XL581; AZD8330 (ARRY-424704 / ARRY-704); RO5126766 (Roche, PLoS One. 2014 Nov). One or more of the following may be selected: (as described in 25;9(11)) and GSK1120212 (or JTP-74057, as described in Clin Cancer Res. 2011 Mar 1;17(5):989-1000). The MAPK inhibitor may be PLX8394, LXH254, GDC-5573, or LY3009120.

[0115] In some embodiments, the anticancer agent is a disruptor or inhibitor of the RAS-RAF-ERK, PI3K-AKT-TOR, or PI3K-AKT signaling pathway. Such agents are known in the art. The PI3K / AKT inhibitor may include, but is not limited to, one or more PI3K / AKT inhibitors listed in Cancers (Basel) 2015 Sep;7(3):1758-1784. For example, the PI3K / AKT inhibitor may be selected from one or more of NVP-BEZ235;BGT226;XL765 / SAR245409;SF1126;GDC-0980;PI-103;PF-04691502;PKI-587;GSK2126458.

[0116] In some embodiments, the anticancer agent is a PD-1 or PD-L1 antagonist. Such agents are known in the art. In some embodiments, additional therapeutic agents include ALK inhibitors, HER2 inhibitors, EGFR inhibitors, IGF-1R inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, and immunotherapies. In some embodiments, additional therapeutic agents include FGFR inhibitors, PARP inhibitors, BET inhibitors, PRMT5i inhibitors, MAT2A inhibitors, VEGF inhibitors, and HDAC inhibitors. In some embodiments, the therapeutic agent may be a pan-RTK inhibitor (such as afatinib).

[0117] IGF-1R inhibitors are known in the art and include lincitinib or its pharmaceutically acceptable salts. EGFR inhibitors are known in the art and include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®), panitumumab (Vectibix®), zaltumumab, nimotuzumab, and matuzumab. Further antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi et al., Br. J. Cancer 1993, 67:247-253; Teramoto et al., Cancer 1996, 77:639-645; Goldstein et al., Clin. Cancer Res. 1995, 1:1311-1318; Huang et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang et al., Cancer Res. 1999, 59:1236-1243. The EGFR inhibitor may be a monoclonal antibody Mab E7.6.3 (Yang, 1999 (see above)), or Mab C225 (ATCC accession number HB-8508), or an antibody or antibody fragment having binding specificity thereto.

[0118] Examples of small molecule antagonists of EGFR include gefitinib (Iressa®), erlotinib (Tarceva®), and lapatinib (TykerB®). See, for example, Yan et al., Pharmacogenetics and Pharmacogenomics in Oncology Therapeutic Antibody Development, BioTechniques 2005, 39(4):565-8; and Paez et al., EGFR Mutations in Lung Cancer Correlation with Clinical Response to Gefitinib Therapy, Science 2004, 304(5676):1497-500. In some embodiments, the EGFR inhibitor is osimertinib (Tagrisso®). Further non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and any pharmaceutically acceptable salts of such EGFR inhibitors: EP0520722;EP0566226;WO96 / 33980;U.S. Patent No. 5,747,498;WO96 / 30347;EP0787772;WO97 / 30034;WO97 / 30044;WO97 / 38994;WO97 / 49688;EP837063;WO98 / 02434;WO97 / 38983;WO95 / 19774;WO95 / 19970;WO97 / 13771;WO98 / 02437;WO98 / 0243 8;WO97 / 32881;DE19629652;WO98 / 33798;WO97 / 32880;WO97 / 32880;EP682027;WO97 / 02266;WO97 / 27199;WO98 / 07726;WO97 / 34895;WO96 / 31510;WO98 / 14449;WO98 / 144 50; WO98 / 14451; WO95 / 09847; WO97 / 19065; WO98 / 17662; US Patent No. 5,789,427; US Patent No. 5,650,415; US Patent No. 5,656,643; WO99 / 35146; WO99 / 35132; WO99 / 07701; and WO92 / 20642.Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler et al., Exp. Opin. Ther. Patents 1998, 8(12):1599-1625. In some embodiments, EGFR inhibitors are ERBB inhibitors. In humans, the ERBB family includes HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3), and HER (ERBB4).

[0119] MEK inhibitors are known in the art and include, but are not limited to, pimacertib, selumetinib, cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib (Mektovi®). In some embodiments, the MEK inhibitor targets a MEK mutation that is a class I MEK1 mutation selected from D67N;P124L;P124S; and L177V. In some embodiments, the MEK mutation is a class II MEK1 mutation selected from ΔE51-Q58;ΔF53-Q58;E203K;L177M;C121S;F53L;K57E, Q56P; and K57N.

[0120] PI3K inhibitors are known in the art, including wortmannin; 17-hydroxywortmannin analog described in WO06 / 044453; 4-[2-(1H-indazole-4-yl)-6-[[4-(methylsulfonyl)piperazine-1-yl]methyl]thieno[3,2-d]pyrimidine-4-yl]morpholine (also known as pictilisib or GDC-0941, described in WO09 / 036082 and WO09 / 055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinoline-3-yl)-2,3-dihydroimidazo[4,5-c]quinoline-1-yl]phenyl]propionitrile (BEZ 235 or NVP-BEZ Also known as 235 and described in WO06 / 122806); (S)-l-(4-((2-(2-aminopyrimidine-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in WO08 / 070740); LY294002 (2-(4-morpholinyl)-8-phenyl-4H-l-benzopyran-4-one (available from Axon Medchem); PI 103 hydrochloride (3-[4-(4-morpholinylpyrido[3',2':4,5]fl[3,2-d]pyrimidine-2-yl]phenol hydrochloride (available from Axon Medchem); PIK 75(2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridine-3-yl)methylene]-1-methylhydrazide-benzenesulfonic acid, monohydrochloride) (available from Axon Medchem); PIK 90(N-(7,8-dimethoxy-2,3-dihydroimidazo[l,2-c]quinazolin-5-yl)nicotinamide (available from Axon Medchem); AS-252424(5-[l-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidined-2,4-dione (Axon Available from Medchem); TGX-221(7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrinidine-4-one (available from Axon Medchem); XL-765;Other PI3K inhibitors include, but are not limited to, XL-147 and CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI 00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0121] AKT inhibitors are known in the art, including Akt-1-1 (inhibits Aktl) (Barnett et al., Biochem. J.2005,385(Pt.2):399-408); Akt-1-1,2 (inhibits Akl and 2) (Barnett et al., Biochem. J.2005,385(Pt.2):399-408); API-59CJ-Ome (e.g., Jin et al., Br. J. Cancer 2004,91:1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05 / 011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963; Sarkar and Li J Nutr.2004,134(12)). Examples include, but are not limited to, Suppl):3493S-3498S); Perifosine (e.g., interfering with Akt membrane localization; Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15):5242-52); Phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis Expert. Opin. Investig. Drugs 2004, 13:787-97); and trisirivine (TCN or API-2 or NCI discriminant: NSC 154020; Yang et al., Cancer Res. 2004, 64:4394-9).

[0122] mTOR inhibitors are known in the art, including ATP-competitive mTORC1 / mTORC2 inhibitors, such as PI-103, PP242, PP30; and Torin. 1; FKBP12 enhancers; 4H-1-benzopyran-4-one derivatives; and temsirolimus (Torisel®); everolimus (Afinitor®; WO94 / 09010); ridafololimus (also known as deforolimus or AP23573); rapalogs, e.g., those disclosed in WO98 / 02441 and WO01 / 14387, e.g., AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779); 40-epi-(tetrazolite)-rapamycin (also known as ABT578); 32-deoxorapamycin; 16-pentinyloxy-32(S)-dihydrorapamycin Herring; derivatives disclosed in WO05 / 005434; U.S. Patents 5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, 5,120,842, and 5,256,790, as well as WO94 / 090101, WO92 / 05179, WO93 / 111130, WO94 / 02136, WO Examples of mTOR inhibitors include, but are not limited to, derivatives disclosed in 94 / 02485, WO95 / 14023, WO94 / 02136, WO95 / 16691, WO96 / 41807, WO96 / 41807, and WO2018204416, as well as rapamycin (also known as sirolimus) and its derivatives, including phosphorus-containing rapamycin derivatives (e.g., WO05 / 016252). In some embodiments, the mTOR inhibitor is a bisteric inhibitor having the following structure, such as RMC-5552 (see, for example, WO2018204416, WO2019212990, and WO2019212991).

[0123] [ka]

[0124] BRAF inhibitors that can be used in combination with the compounds of the present invention are known in the art and include, for example, vemurafenib, dabrafenib, and encorafenib. BRAF may include class 3 BRAF mutations. In some embodiments, class 3 BRAF mutations are selected from one or more of the following amino acid substitutions in human BRAF: D287H;P367R;V459L;G466V;G466E;G466A;S467L;G469E;N581S;N581I;D594N;D594G;D594A;D594H;F595L;G596D;G596R, and A762E.

[0125] MCL-1 inhibitors are known in the art and include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the major anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression and resistance not only to conventional chemotherapy but also to targeted therapies, including BCL-2 inhibitors such as ABT-263.

[0126] In some embodiments, additional therapeutic agents are SHP2 inhibitors. SHP2 inhibitors are known in the art. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene, which contributes to several cellular functions, including proliferation, differentiation, maintenance of the cell cycle, and migration. SHP2 has two N-terminal Src homology domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. The two SH2 domains regulate the intracellular localization and functional regulation of SHP2. The molecule exists in an inactive, self-inhibitory conformation stabilized by a binding network involving residues from both the N-SH2 and PTP domains. For example, stimulation by cytokines or growth factors acting via receptor tyrosine kinases (RTKs) results in exposure of the catalytic site, leading to enzymatic activation of SHP2.

[0127] SHP2 is involved in signaling via the RAS mitogen-activated protein kinase (MAPK), JAK-STAT, or phosphoinositol 3-kinase-AKT pathways. Mutations in the PTPN11 gene and subsequent SHP2 mutations have been identified in several human developmental disorders, including Noonan syndrome and Leopard syndrome, as well as in human cancers such as juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung, and colon cancer. Some of these mutations destabilize the autoinhibitory conformation of SHP2, promoting autoactivation or enhanced growth factor-driven activation of SHP2. Therefore, SHP2 is a very attractive target for the development of novel therapies for the treatment of various diseases, including cancer. SHP2 inhibitors (e.g., RMC-4550 or SHP099) combined with RAS pathway inhibitors (e.g., MEK inhibitors) have been shown to inhibit the growth of several cancer cell lines (e.g., pancreatic, lung, ovarian, and breast cancer) in vitro. Therefore, combination therapy using SHP2 inhibitors and RAS pathway inhibitors can be a common strategy for preventing tumor resistance in a wide range of malignancies.

[0128] Such non-limiting examples of SHP2 inhibitors are known in the art, including: Chen et al. Mol Pharmacol. 2006, 70, 562; Sarver et al., J.Med.Chem. 2017, 62, 1793; Xie et al., J.Med.Chem. 2017, 60, 113734; and Igbe et al. al., Oncotarget, 2017, 8, 113734; and patent applications: WO2023282702, WO2023280283, WO2023280237, WO2023018155, WO2023011513, WO2022271966, WO2022271964, WO2022271911, WO2022259157, WO2022242767, WO2022241975, WO2022237676, WO2022237367, WO2022237178, WO2022235822, WO20222084008, WO2022135568, WO2021176072, WO2021171261, WO2021149817, WO2021148010 , WO2021147879, WO2021143823, WO2021143701, WO2021143680, WO2021121397, WO2021119525, WO2021115286, WO2021110796 , WO2021088945, WO2021073439, WO2021061706, WO2021061515, WO2021043077, WO2021033153, WO2021028362, WO2021033153 , WO2021028362, WO2021018287, WO2020259679, WO2020249079, WO2020210384, WO2020201991, WO2020181283, WO2020177653 , WO2020165734, WO2020165733, WO2020165732, WO2020156243, WO2020156242, WO2020108590, WO2020104635, WO2020094104 , WO2020094018, WO2020081848, WO2020073949, WO2020073945, WO2020072656, WO2020065453, WO2020065452, WO2020063760,WO2020061103、WO2020061101、WO2020033828、WO2020033286、WO2020022323、WO2019233810、WO2019213318、WO2019183367、WO2019183364、WO2019182960、WO2019167000、WO2019165073、WO2019158019、WO2019152454、WO2019051469、WO2019051084、WO2018218133、WO2018172984、WO2018160731、WO2018136265、WO2018136264、WO2018130928、WO2018129402、WO2018081091、WO2018057884、WO2018013597、WO2017216706、WO2017211303、WO2017210134、WO2017156397、WO2017100279、WO2017079723、WO2017078499、WO2016203406、WO2016203405、WO2016203404、WO2016196591、WO2016191328、WO2015107495、WO2015107494、WO2015107493、WO2014176488、WO2014113584、CN115677661、CN115677660、CN115611869、CN115521305、CN115490697、CN115466273、CN115394612、CN115304613、CN115304612、CN115300513、CN115197225、CN114957162、CN114920759、CN114716448、CN114671879、CN114539223、CN114524772、CN114213417、CN114195799、CN114163457、CN113896710、CN113248521、CN113248449、CN113135924、CN113024508、CN112920131、CN112823796、CN112409334、CN112402385、CN112174935、111848599、CN111704611、CN111393459、CN111265529、CN110143949、CN108113848、US11179397、Examples include US11044675, US11034705, US11033547, US11001561, US10988466, US10954243, US10934302, or US10858359, or their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers, each of which is incorporated herein by reference.

[0129] In some embodiments, the SHP2 inhibitor binds to the active site. In some embodiments, the SHP2 inhibitor is a mixed-type irreversible inhibitor. In some embodiments, the SHP2 inhibitor binds to the allosteric site, for example, a non-covalent allosteric inhibitor. In some embodiments, the SHP2 inhibitor is a covalent SHP2 inhibitor, such as an inhibitor that targets a cysteine ​​residue (C333) outside the active site of the phosphatase. In some embodiments, the SHP2 inhibitor is a reversible inhibitor. In some embodiments, the SHP2 inhibitor is an irreversible inhibitor. In some embodiments, the SHP2 inhibitor is SHP099. In some embodiments, the SHP2 inhibitor is TNO155 having the following structure:

[0130] [ka]

[0131] Alternatively, it may be a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RMC-4550. In some embodiments, the SHP2 inhibitor is RMC-4630 having the following structure:

[0132] [ka]

[0133] Alternatively, it may be a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3068 having the following structure.

[0134] [ka]

[0135] Alternatively, it may be a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3312. In some embodiments, the SHP2 inhibitor is the following compound,

[0136] [ka]

[0137] Alternatively, it may be a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RLY-1971 having the following structure.

[0138] [ka]

[0139] Alternatively, it may be a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is ERAS-601. In some embodiments, the SHP2 inhibitor is BBP-398.

[0140] In some embodiments, additional therapeutic agents are selected from the group consisting of MEK inhibitors, HER2 inhibitors, SHP2 inhibitors, CDK4 / 6 inhibitors, mTOR inhibitors, SOS1 inhibitors, and PD-L1 inhibitors. See, for example, Hallin et al., Cancer Discovery, DOI:10.1158 / 2159-8290 (October 28, 2019) and Canon et al., Nature, 575:217 (2019). In some embodiments, the Ras inhibitor of the present invention is used in combination with a MEK inhibitor and an SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and an SOS1 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a PD-L1 inhibitor and an SHP2 inhibitor. In some embodiments, the Ras inhibitor of the present invention is used in combination with a MEK inhibitor and an SHP2 inhibitor. In some embodiments, the Ras inhibitors of the present invention are used in combination with SHP2 inhibitors and Ras inhibitors that inhibit multiple Ras isoforms and / or variants. In some embodiments, the cancer is lung cancer, and the treatment comprises administering the Ras inhibitors of the present invention in combination with a second or third therapeutic agent, such as an SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or variants. In some embodiments, the cancer is colorectal cancer, and the treatment comprises administering the Ras inhibitors of the present invention in combination with a second or third therapeutic agent (e.g., an SHP2 inhibitor and a Ras inhibitor that inhibits multiple Ras isoforms and / or variants). In some embodiments, the Ras inhibitors of the present invention are used in combination with immunotherapy and optionally with chemotherapeutic agents.

[0141] Proteasome inhibitors are known in the art and include, but are not limited to, carfilzomib (Kyprolis®), bortezomib (Velcade®), and oprozomib.

[0142] Immunotherapy includes, but is not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically modified T cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTE), and anti-PD-1, anti-PD-L1, anti-CTLA4, anti-LAGl, and anti-OX40 agents. Other immunotherapies are known in the art.

[0143] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that modulate the immune response) that contain an imide group. Examples of IMiDs include thalidomide and its analogues (lenalidomide, pomalidomide, and apremilast).

[0144] Exemplary anti-PD-1 antibodies and their uses are described in Goldberg et al., Blood 2007, 110(1):186-192; Thompson et al., Clin. Cancer Res. 2007, 13(6):1757-1761; and WO06 / 121168 A1), and further described elsewhere in this specification.

[0145] FGFR inhibitors are known in the art and include pemigatinib and erdafitinib, which are FGFR2 and FGFR4 inhibitors. See, for example, Cancers (Basel), 2021 Jun;13(12)2968.

[0146] BET inhibitors, such as romidepsin, panobinostat, and bellinostat, are known in the art. See, for example, British J. Cancer 124:1478 (2021).

[0147] PRMT5i inhibitors, such as PF-0693999, PJ-68, and MRTX1719, are known in the art. See, for example, Biomed. Pharmacotherapy 144:112252 (2021).

[0148] MAT2A inhibitors, such as AG-270 and IDE397, are known in the art. See, for example, Exp Opin Ther Patents (2022) DOI:10.1080 / 13543776.2022.2119127.

[0149] GITR agonists include GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), e.g., GITR fusion proteins described in U.S. Patent No. 6,111,090, No. 8,586,023, WO2010 / 003118, and WO2011 / 090754; or e.g., U.S. Patent No. 7,025,962, EP1947183, U.S. Patent No. 7,812,135, U.S. Patent No. 8,388,967, U.S. Patent No. 8,59 Examples of anti-GITR antibodies include, but are not limited to, those described in U.S. Patent No. 1,886, U.S. Patent No. 7,618,632, EP1866339, and WO2011 / 028683, WO2013 / 039954, WO05 / 007190, WO07 / 133822, WO05 / 055808, WO99 / 40196, WO01 / 03720, WO99 / 20758, WO06 / 083289, WO05 / 115451, and WO2011 / 051726.

[0150] Another example of a therapeutic agent that may be used in combination with the crystalline compounds of the present invention is an anti-angiogenic agent. Anti-angiogenic agents are known in the art and include, but are not limited to, chemical compositions, antibodies, antigen-binding domains, radionuclides, and combinations thereof and conjugates, which are synthetically prepared in vitro. Anti-angiogenic agents may be agonists, antagonists, allosteric modulators, toxins, or, more generally, may act to inhibit or stimulate their targets (e.g., by activating or inhibiting receptors or enzymes), thereby promoting cell death or halting cell proliferation. In some embodiments, one or more additional therapies include an anti-angiogenic agent.

[0151] Anti-angiogenic agents may include MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase 11) inhibitors. Non-exclusive examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include arecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors include WO96 / 33172, WO96 / 27583, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, WO90 / 05719, WO99 / 52910, and WO99 / 52 As described in 889, WO99 / 29667, WO99007675, EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578, and US20090012085, and U.S. Patents 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors have little to no activity in inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 or AMP-9 compared to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors are AG-3340, RO 32-3555, and RS 13-0830.

[0152] Further exemplary anti-angiogenic agents include KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., VEGF (e.g., bevacizumab), or antibodies or antigen-binding regions that specifically bind to their soluble VEGF receptors or ligand-binding regions), e.g., VEGF-TRAP®, and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind to them), VEGF inhibitors, EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to them), e.g., Vectibix® (panitumumab), erlotinib (Tarceva®), anti-Angl and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind to them or their receptors, e.g., Tie2 / Tek), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to them). Other anti-angiogenic agents include Canasamin, IL-8, B-FGF, Tek antagonists (US2003 / 0162712; US6,413,932), anti-Tweak agents (e.g., antibodies or antigen-binding domains that specifically bind, or soluble Tweak receptor antagonists; see US6,727,225), ADAM distointegrin domains that antagonize the binding of integrins to their ligands (US2002 / 0042368), and anti-eph receptors or anti-ephrin antibodies or antigens that specifically bind. Examples include binding regions (U.S. Patents No. 5,981,245; No. 5,728,813; No. 5,969,110; No. 6,596,852; No. 6,232,447; No. 6,057,124 and their patent family members), anti-PDGF-BB antagonists (e.g., antibody or antigen binding regions that specifically bind), antibody or antigen binding regions that specifically bind to PDGF-BB ligands, and PDGFR kinase inhibitors (e.g., antibody or antigen binding regions that specifically bind to them). Additional anti-angiogenic agents include SD-7784 (Pfizer, USA); sirengitide (Merck KGaA, Germany, EPO0770622); pegaptanib octasodium (Gilead Sciences, USA);Alpha-statin (BioActa, UK); M-PGA (Celgene, USA, US5712291); Ilostat (Arriva, USA, US5892112); Emaxanib (Pfizer, USA, US5792783); Batalanib (Novartis, Switzerland); 2-Methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); Anecol acetate (Alcon, USA); Alpha-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands), DAC anti-angioplastic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP0970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen-E fragment (BioActa, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); Maspin (Sosei, Japan); 2-Methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IV AX, USA); BeneFin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP02233610); Platelet factor 4 (RepliGen, USA, EP407122); Vascular endothelial growth factor antagonist (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis inhibitor (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA);MAb, alpha-5 beta-3 integrin, second generation (Applied Molecular Evolution, USA and Medlmmune, USA); Enzastaurin hydrochloride (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); rBPI 21 and BPI-derived anti-angiogenic agents (XOMA, USA); PI 88 (Progen, Australia); Silengitide (Merck KGaA, Germany; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); Endostatin (Boston Children's Hospital, USA); ATN 161 (Attenuon, USA); 2-Methoxyestradiol (Boston Children's Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); Batalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); Tissue Factor Pathway Inhibitor (EntreMed, USA); Pegaptanib (Pinn) (Gilead Sciences, USA); Xantollyzole (Yonsei University, South Korea); Vaccine, Gene-Based, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV 5.2 (Supratek, Canada); SDX 103 (University of California at San Diego, USA); PX 478(ProlX,USA);Metastatin (EntreMed, USA); Troponin I (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); o-Guanidine (Dimensional Pharmaceuticals, USA); Motupolamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); Atiprimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); Vaccine, Angiogenesis (EntreMed, USA); Urokinase Plasminogen Activator Inhibitor (Dendreon, USA); Ogluphanide (pINN) (Melmotte, USA); HIF-Ralfa Inhibitor (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); Angiocidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Research Institute of Chemical Technology, South Korea); GW 2286 (GlaxoSmithKline, UK); EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); Drug delivery system, intraocular, 2-methoxyestradiol; Anguinex (Maastricht University, Netherlands, and Minnesota University, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); Tumor necrosis factor-alpha inhibitor; SU 11248 (Pfizer, USA and SUGEN USA); ABT518 (Abbott, USA);YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, Alpha 5 Beta (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson & Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan); Comblestatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); Ilsogladine (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); Squalamine (Genaera, USA); RPI 4610 (Sirna, USA); Heparanase inhibitor (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VE-cadherin-2 antagonist (ImClone Systems, USA);Vasostatin (National Institutes of Health, USA); Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); Sclerated soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand; Examples include Regeneron (USA) and thrombospondin 1 inhibitors (Allegheny Health, Education and Research Foundation, USA).

[0153] Further examples of therapeutic agents that may be used in combination with the compounds of the present invention include agents (e.g., antibodies, antigen-binding domains, or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor), as well as antibodies or antigen-binding domains that specifically bind to the receptor c-Met. Such agents are known in the art.

[0154] Another example of a therapeutic agent that may be used in combination with the compounds of the present invention is an autophagy inhibitor. Autophagy inhibitors are known in the art and include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil®), bafilomycin A1, 5-amino-4-imidazole carboxamidriboside (AICAR), okadaic acid, autophagy-suppressing algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Furthermore, antisense or siRNAs that inhibit the expression of proteins including (but not limited to) ATG5 (which is involved in autophagy) may be used. In some embodiments, one or more additional therapies include an autophagy inhibitor.

[0155] Another example of therapeutic agents that can be used in combination with the crystalline compounds of the present invention is antineoplastic agents, which are known in the art. In some embodiments, one or more additional therapies include antineoplastic agents. Non-limiting examples of antineoplastic agents include acemannan, acralubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amiphostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, algravin, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, bromodeoxyuridine, capecitabine, cermoloukin, cetrorelix, cladribine, clotrimazole, cytarabine ocphosphate, and DA3030 (Dong-A), daclizumab, denileukin difutitox, deslorerin, dexrazoxane, dilazep, docetaxel, docosanol, doxelcalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, ederfosine, edrecolomab, eflornithine, emiteflu, epirubicin, epoetin beta, etoposide phosphate, exemestane, exislind, fadrozol, filgrastim, finasteride, fludarabine phosphate, formestan, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil / oteracil / tegafur combination, Glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin (imiquimod), interferon alpha, interferon alpha (natural type), interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-NI, interferon alpha-n3, interferon alpha-con-1, interferon alpha (natural type), interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, natural type interferon gamma-la, interferon gamma-lb, interleukin-1 beta, iobenguan, irinotecan, ilsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprorelin, levamizole + fluorouracil, rialozol, lovaplatin, lonidamin, lovastatin, masopropyl, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, mispaired double-stranded RNA, mitogluzone, mitractol, mitoxantrone, morglamostim, nafarelin, naloxone + pentazocine, naltograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-promoting protein, NSC631570 Octreotide, Oprelbequine, Osateron, Oxaliplatin, Paclitaxel, Pamidronic acid, Pegaspargase, Peginterferon alpha-2b, Pentosan, Sodium polysulfate, Pentostatin, Picibanil, Pirarubicin, Rabbit anti-thymocyte polyclonal antibody, Polyethylene glycol interferon alpha-2a, Porfimer sodium, Raloxifene, Larcitrexed, Rasburiembodiment, Rhenium etidronate Re 186, RII retinamide, Rituximab, Romultide, Samarium (153 Sm) Lexidrona, Salglamostim, Schizophyllan, Sobuzoxane, Sonelmin, Strontium-89 Chloride, Suramin, Tasonelmin, Tazarotene, Tegafur, Temoporfin, Temozolomide, Teniposide, Tetrachlorodecaoxide, Thalidomide, Thimalfacin, Thyroid-stimulating hormone alpha, Topotecan, Toremifene, Tositumomab-iodine-131, Trastuzumab, Treosulfan, Tretinoin, Trilostane, Trimethrexate, Triptorelin, Tumor necrosis factor alpha, Natural type, Ubenimex, Bladder cancer vaccine, Maruyama vaccine, Melanoma solubilin vaccine, Barrubicin, Verteporfin, Vinorelbine, Birulizine, Dinostatin stimalamer, or Zoledronic acid; Abarelix; AE 941 (Aeterna), Ambamustine, Antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), Decitabine, Dexaaminoglutethimide, Diadicon, EL 532 (Elan), EM 800 (Endorecherche), Enyluracil, Etanidazole, Fenretinide, Filgrastim SD01 (Amgen), Fulvestrant, Gallocitabine, Gastrin 17 Immunogen, HLA-B7 gene therapy (Vical), Granulocyte-macrophage colony-stimulating factor, Histamine dihydrochloride, Ibritumomab tiuxetan, Ilostat, IM 862 (Cytran), Interleukin-2, Iproxyfen, LDI 200 (Milkhaus), Religistim, Lintuzumab, CA 125 MAb (Biomira), Cancer MAb (Japan PharmaceuticalDevelopment), HER-2 and Fc MAb (Medarex), Idiotype 105AD7 MAb (CRC Technology), Idiotype CEA MAb (Trilex), LYM-1-iodine-131 MAb (Techniclone), Polymorphoemic mucin-yttrium-90 MAb (Antisoma), Marimast, Menogalil, Mitsumomab, Motexafingadolinium, MX 6 (Galderma), Nelarabine, Noratexed, P30 protein, Pegvisomant, Pemetrexed, Porphyromycin, Prinomast, RL 0903 (Shire), Rubitecan, Satraplatin, Sodium phenylacetate, Sparphosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA Examples include 077 (Tanabe), tetrathiomolybdate, saliblastin, thrombopoietin, tin-ethylethiopurine, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysis product vaccine (New York Medical College), viral melanoma cell solubilized vaccine (Royal Newcastle Hospital), or valspodar.

[0156] Further examples of therapeutic agents that can be used in combination with the crystalline compounds of the present invention include ipilimumab (Yervoy®); tremelimumab; galiximab; nivolumab, also known as BMS-936558 (Opdivo®); pembrolizumab (Keytruda®); avelumab (Bavencio®); AMP224; BMS-936559; MPDL3280A (also known as RG7 446);MEDI-570;AMG557;MGA271;IMP321;BMS-663513;PF-05082566;CDX-1127;Anti-OX40 (Providence Health Services);huMAbOX40L;Atasicept;CP-870893;Lucatumumab;Dasetuzumab;Muromonab-CD3;Ipilumumab;MEDI4736 (Imfinzi (registered trademark));MSB0010718C;AMP 224; adalimumab (Humira®); ado-trastuzumab emtansine (Kadcyla®); aflibercept (Eylea®); alemtuzumab (Campath®); basiliximab (Simulect®); belimumab (Benlysta®); basiliximab (Simulect®); belimumab (Benlysta®); brentuximab vedotin (Adcetris®); canakinumab (Ilaris®); certolizumab pegol (Cimzia®); daclizumab (Zenapax®); daratumumab (Darzalex®) (Registered Trademark)); Denosumab (Prolia(Registered Trademark)); Eculizumab (Soliris(Registered Trademark)); Efalizumab (Raptiva(Registered Trademark)); Gemtuzumab Ozogamicin (Mylotarg(Registered Trademark)); Golimumab (Simponi(Registered Trademark)); Ibritumomab Chiuxetan (Zevalin(Registered Trademark)); Infliximab (Remicade(Registered Trademark)); Motavizumab (Numax(Registered Trademark)); Natalizumab (Tysabri(Registered Trademark)); Obinutuzumab (Gazyva(Registered Trademark)); Ofatumumab (Arzerra(Registered Trademark)); Omalizumab (Xolair(Registered Trademark)); Palivizumab (Synagis(Registered Trademark));Examples include pertuzumab (Perjeta®); pertuzumab (Perjeta®); ranibizumab (Lucentis®); laxibamumab (Abthrax®); tocilizumab (Actemra®); tositumomab; tositumomab-i-131; tositumomab and tositumomab-i-131 (Bexxar®); ustekinumab (Stelara®); AMG 102; AMG 386; AMG 479; AMG 655; AMG 706; AMG 745; and AMG 951.

[0157] The crystalline compounds described herein can be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Therefore, in some embodiments, one or more compounds of this disclosure are co-administered with other therapies as described herein. When used in combination therapy, the compounds described herein may be administered simultaneously with or separately from a second agent. This combined administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the crystalline compounds described herein and any agent described herein may be formulated together in the same dosage form and administered simultaneously. Alternatively, the crystalline compounds of the present invention and any therapy described herein may be administered simultaneously, and both agents may exist in separate formulations. In another alternative, the crystalline compounds of this disclosure may be administered, followed by any therapy described herein, or vice versa. In some embodiments of separate administration protocols, the crystalline compounds of the present invention and any therapy described herein are administered at intervals of minutes, hours, or days.

[0158] In some embodiments of any of the methods described herein, the first therapy (e.g., the compounds of the present invention) and one or more additional therapies are administered simultaneously or sequentially in any order. The first therapeutic agent may be administered immediately before or after one or more additional therapies, or up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours before or after, or up to 1-7, 1-14, 1-21, or 1-30 days before or after.

[0159] The present invention also features a kit comprising (a) a pharmaceutical composition comprising an agent described herein (e.g., a crystalline compound of the present invention), and (b) a package insert containing instructions for carrying out any of the methods described herein. In some embodiments, the kit comprises (a) a pharmaceutical composition comprising an agent described herein (e.g., a crystalline compound of the present invention), (b) one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents), and (c) a package insert containing instructions for carrying out any of the methods described herein.

[0160] One aspect of the present invention, intending to treat a disease or associated symptoms by a combination of pharmaceutically active compounds that can be administered separately, further relates to combining separate pharmaceutical compositions in the form of a kit. The kit may comprise two separate pharmaceutical compositions, namely the crystalline compounds of the present invention, and one or more additional therapies. The kit may comprise containers for housing the separate compositions, such as divided bottles or divided foil packets. Additional examples of containers include syringes, boxes, and bags. In some embodiments, the kit may comprise instructions for using the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), administered at different dosing intervals, or when titration of the individual components of the combination is desired by the prescribing healthcare professional.

[0161] The following embodiments illustrate the present invention further, but should not be construed as limiting its scope. [Examples]

[0162] This disclosure is further illustrated by the following examples and synthesis examples, which should not be considered to limit the scope or spirit of this disclosure to the specific procedures described herein. It should be understood that the examples are provided to illustrate specific embodiments and are not intended to imply any limitation on the scope of this disclosure. It should also be understood that various other embodiments, modifications, and means to which equivalents thereof may be utilized, without departing from the spirit of this disclosure or the appended claims, which themselves may be suggested to those skilled in the art.

[0163] Example 1 This example illustrates an exemplary method for preparing a free base of compound A in crystalline form 1, according to one embodiment of the present invention.

[0164] In one method, approximately 50 mg of the free base of amorphous compound A was dissolved in a minimal amount of MeOH at ambient temperature (approximately 20-25°C). Four times the volume of water was quickly added to the resulting clear solution to form a precipitate. The precipitate was isolated by centrifugation at 14,000 rpm using a 0.45 μm nylon membrane filter to obtain crystalline form 1 of the free base of compound A.

[0165] Alternatively, 1 mL of a saturated aqueous solution of compound A at 50°C was added to 1 mL of methanol. The turbid solution was cooled to ambient temperature to obtain crystalline form 1 of the free base of compound A. Crystals deemed to be of sufficient size and quality for single-crystal X-ray diffraction analysis were taken and characterized by a crystallographer.

[0166] Alternatively, a mixture of 201.3 mg of amorphous compound A and 2.5 mL of aqueous acetate buffer (pH 4) was stirred at ambient temperature for approximately 7 days. The mixture was centrifuged, the liquid was decanted, and the remaining solid was dried in air to obtain crystalline form 1 of the free base of compound A.

[0167] Alternatively, approximately 50 mg of amorphous compound A was dissolved in a minimal amount of acetone at ambient temperature (e.g., approximately 20-25°C) to form a clear solution. Water was added to this solution until a large amount of solid precipitated (approximately 1:1 (v / v) acetone / water). The solid was recovered by centrifugation at 14,000 rpm through a 0.45 μm nylon membrane filter to obtain crystalline form 1.

[0168] Alternatively, 9.5 g of amorphous compound A was added to 38 mL of MeOH. The mixture was filtered to obtain a clear solution. The clear solution was quickly added to 152 mL of water to form a suspension. Form 1 seeds were added to the suspension, and the mixture was stirred at 25°C for 24 hours, then at 5°C for 24 hours. The solid was recovered by vacuum filtration. After drying the solid at 25°C for about 6 days, 8.2 g of Form 1 was obtained as a white powder (78% yield).

[0169] Alternatively, crystalline form 5 of compound A (see Example 13) spontaneously converts to crystalline form 1 of compound A when crystalline form 5 is exposed to a water activity of 0.4 or higher at 25°C.

[0170] Example 2 This embodiment demonstrates the characterization of crystalline form 1 of compound A free base by X-ray powder diffraction (XRPD) according to one embodiment of the present invention.

[0171] XRPD measurements were performed using one of the two RigakuSmart-Lab X-ray diffraction systems. Each was set to a Bragg-Brentano reflection configuration using a line-source X-ray beam. The Bragg-Brentano configuration was controlled by passive divergent and accepting slits, with the sample itself acting as a focusing element of the optical system. Powder samples were prepared in a low-background Si holder using light manual pressure to keep the sample surface flat and at the same height as the reference plane of the sample holder. The specifications of the diffractometer and data acquisition parameters for diffraction system 1 are shown in Table 1. The specifications of the diffractometer and data acquisition parameters for diffraction system 2 are shown in Table 2. The XRPD of crystalline form 1 of compound A as a mixed methanol is shown in Figure 1A. In the essentially pure material of crystalline form 1 as a mixed solvate of methanol and water, a peak can be observed at a refraction angle of 2θ, as shown in Tables 3 and 4 (runs 1 and 2, respectively). The XRPD of crystalline form 1 as a trihydrate solvate is shown in Figure 1B. In the essentially pure material of crystalline form 1 as a trihydrate solvate, a peak can be observed at a refraction angle of 2θ, as shown in Table 5.

[0172] [Table 1]

[0173] [Table 2]

[0174] [Table 3-1]

[0175] [Table 3-2]

[0176] [Table 4-1]

[0177] [Table 4-2]

[0178] [Table 4-3]

[0179] [Table 5-1]

[0180] [Table 5-2]

[0181] Example 3 This embodiment demonstrates the characterization of crystalline form 1 of the free base of compound A by single-crystal X-ray crystal structure analysis according to one embodiment of the present invention. The X-ray crystal structure of crystalline form 1 (asymmetric unit) of the free base of compound A is shown in Figure 2. This structure was determined to be the free base of compound A as a mixed solvate of methanol and water.

[0182] Select a portion of the free base of compound A and use formula 2(C 44 H 58A pale brown, plate-like crystal with approximately dimensions of 0.42 × 0.19 × 0.04 mm, containing N8O5S)·5.55(H2O)·2.19(CH4O), was mounted randomly oriented on a Mitegen micromesh mount. Preliminary inspection and data acquisition were performed using Cu Kα radiation (λ=1.54178A) on a Bruker AXS D8 Quest CMOS diffractometer equipped with a 4-axis copper stage, I-μ-S microsource X-ray tube, transversely tilted multilayer optics, PhotonIII-C14 single-photon count detector, and Oxford Cryosystems cryogenic apparatus. The initial unit cell was subjected to Apex3 v2019.11-0 at a temperature of 150K. 3 We used [tool name] to make decisions and collect data. We used SAINT V8.40B to integrate the frames. 3 A total of 59,167 reflections were collected, of which 19,822 were unique. Cell constants for data collection were obtained from least-squares refinement using 9,955 reflections in the range of 4.1903 to 78.2402°. The parameters and calculated volumes of the triclinic cell were a=10.8182(3)A, b=13.3787(4)A, c=17.6881(5)A, α=88.4427(15)°, β=79.5367(16)°, γ=82.0377(16) and °V=2493.21(12)A 3 Therefore, when Z=1 and the formula weight is 1792.50, the calculated density is 1.194 g / cm³. 3 The linear absorption coefficient is 1.055 / mm for Cu Kα rays. Scaling and SADABS 2016-2 4 Multi-scan absorption correction was applied using [a specific method / tool]. The transmission coefficient was in the range of 0.6221 to 0.7543. The intensity of equivalent reflection was not averaged during data processing.

[0183] The space group was determined by the XPREP program, which is integrated into SHELXTL. 5 The intensity statistics showed the space group P1(#1). SHELXS 6a The structure was analyzed using a direct method employing SHELXL-2018. 6 and the graphical user interface ShelXle 7Using all reflections, F 2 The structure was refined using the complete matrix least squares method. Additional atoms were placed in the subsequent difference Fourier synthesis. The structure was refined using the complete matrix least squares method, and in the equation, the minimized function is Σw(|F o | 2 - |F c | 2 ) 2 And the weight w is w = 1 / [σ²(F o 2 ) + (0.0831P) 2 It is defined as + 0.3910P], where P=(F o 2 + 2F c 2 The result is ) / 3). The scattering factors were obtained from the International Tables on Crystallography (Vol C Tables 4.2.6.8 and 6.1.1.4). A total of 19,822 independent reflections were used for refinement. A total of 7,187 reflections with F2 > 2σ(F2) were used for the calculation of R1.

[0184] The hydrogen atoms bonded to carbon were geometrically arranged and constrained to rest on the parent atom. The carbon-hydrogen bond distance was constrained to 0.95A in aromatic and alkene CH moieties, and to 1.00, 0.99, and 0.98A in aliphatic CH, CH2, and CH3 moieties, respectively. 2For hybrid nitrogen atoms, the nitrogen-hydrogen bond distance was constrained to 0.88A. The oxygen-hydrogen distance of alcohols was constrained to 0.84A. Hydrogen atoms on the methyl and alcohol moieties were initially allowed to rotate to best fit the experimental electron density. Where necessary, the positions of the H atoms in alcohols were further constrained based on hydrogen bonding considerations. Some H atoms in disordered methyl groups were set to be in alternating positions in the final refinement cycle. The hydrogen atoms of pyramidal R2NH groups were refined, and the NH distance was constrained to 0.88(2)A. The positions of the H atoms in water were initially refined, and the OH and H…H distances were constrained to 0.84(2) and 1.36(2)A, respectively. Where necessary, the positions of the H atoms in water were further constrained based on hydrogen bonding considerations. In the final refinement cycle, the positions of the hydrogen atoms in water and alcohols were set to rest on the positions of their carrier O atoms. iso (H) value U eq The values ​​were set to multiples of (C), with 1.5 for OH and CH3 units and 1.2 for CH and CH2 units, respectively.

[0185] Disorder was observed in some parts of both crystallographically independent molecules. Further and more widespread disorder was observed in the intervening solvate molecules (methanol and water). Disorder was observed in the methylcyclopropane-carboxamide group in molecules A and B. In molecule B, this disorder extends to the main molecule containing the thiazole and hexahydropyridazine rings. In molecule B, the piperazine ring is also disordered.

[0186] For all disordered segments, equivalent portions of both molecules were constrained to have similar arrangements. The Uij components of the atomic displacement parameter (ADP) for disordered atoms closer to each other than 2.0A were constrained to be similar. Under these conditions, the refined occupancy ratios were 0.593(17) to 0.407(17) for the methylcyclopropane-carboxamide group in molecule A, 0.612(2) to 0.388(2) for the methylcyclopropane-carboxamide / thiazole / hexahydropyridazine disorder in molecule B, and 0.681(11) to 0.319(11) for the piperazine disorder in molecule B.

[0187] The oxygen atom of the methanol molecule (O13, C47) was hydrogen-bonded to either methoxy O4B or pyridyl N5B and refined as disordered. Carbon atoms were excluded from disorder. The distance of the CO bonds was constrained to be similar to that of each other and similar to the distance of two other partially occupied methanol molecules. The Uij components of ADP of disordered atoms closer to each other than 2.0A were constrained to be similar. The positions of the hydroxyl H atoms in small portions were constrained based on the consideration of hydrogen bonding. Under these conditions, the occupancy ratio was refined to 0.789(13) to 0.211(13).

[0188] The encapsulated site by the major molecule was refined into two partially occupied water molecules (O12B, O12D). The Uij components of the ADP of the two oxygen atoms were constrained to be similar. The occupancy was refined to 0.171(11) and 0.292(13).

[0189] The solvate molecule was refined as disorder between methanol (O2, C1) and water, with a water molecule and one H atom shared between the two molecules. Adjacent water molecules were included in the disorder, sharing a methanol / water occupancy ratio. The disorder ratio was refined to 0.550(4)–0.450(4) in favor of methanol.

[0190] Adjacent to this disorder, and related to the disorder of methylcyclopropane-carboxamide in both molecules, is further disorder of water and methanol molecules. The disorder is widespread and partially unexplained. Extensive overlap between organic fragments and solvate molecules, unexplained disorder within the disorder, and variations in the occupancy rates of adjacent solvate molecules prevented the construction of a clear and consistent disorder model. The disorder was approximated using partial occupancy. Where possible, the occupancy rates of overlapping portions were constrained to sum to 1, but some unexplained density differences remained that could not be modeled.

[0191] Two disordered methanol molecules were refined as disordered and partially occupied (O8B, C45B; O8D C45D). The distance of the CO bond was constrained to be similar to that of the other disordered methanol molecules (see above). The Uij components of ADP for disordered atoms closer to each other than 2.0A were constrained to be similar. The occupancy was refined to 0.251(11)(O8B) and 0.388(2)(O8D). The occupancy of disordered water molecules refined to the following values: • O7B: 0.612(2) (Same as the major disordered part of methylcyclopropane-carboxamide in molecule B) • O9B1: 0.450 (4) (Same as the water content of the undisordered methanol / water in O2) • O9B2: 0.162(4) (Shares the same location as O9D) • O10B: 0.489 (17) (partially occupied area) • O10C: 0.388(19) (Same as the small, disordered portion of methylcyclopropane-carboxamide in molecule B) • O7D: 0.169 (13) (Partially occupied area) • O9D: 0.388(2) (shares the same location as O9B2).

[0192] The final refinement cycle, involving 1,598 variable parameters and 1,445 constraints, converged with the following unweighted and weighted coincidence coefficients (the maximum parameter shift was 0.007 times its standard uncertainty): R1 = Σ|F o |-|F c | / Σ|F o |=0.0483 wR2={Σ[w(F o 2 -F c 2 )2] / Σ[w(F o 2 ) 2 ]} 0.5 = 0.1381.

[0193] The goodness-of-fit parameter was 1.031. The highest peak in the final difference Fourier map had a height of 0.693 e / A3. The lowest negative peak had a height of -0.271 e / A3.

[0194] The crystal data and data acquisition parameters are shown in Table 6.

[0195] [Table 6-1]

[0196] [Table 6-2]

[0197] Example 4 This embodiment demonstrates the characterization of the crystalline form of compound A by differential scanning calorimetry (DSC) according to one embodiment of the present invention.

[0198] The DSC analysis was performed using a TA Instruments Q2500 Discovery series instrument. The temperature calibration of the instrument was performed using indium. During each analysis, the DSC cell was maintained under a nitrogen purge of approximately 50 mL per minute. The sample was placed in a standard crimped aluminum pan and heated from approximately 25 °C to 350 °C at a rate of 10 °C per minute. The DSC thermogram of crystalline form 1 of compound A free base is shown in Figure 3. The DSC thermogram of crystalline compound A tosylate is shown in Figure 7. The DSC thermogram of crystalline form 2 of compound A is shown in Figure 12. The DSC thermogram of crystalline form 4 of compound A is shown in Figure 16. The DSC thermogram of crystalline form 5 of compound A is shown in Figure 19. The DSC thermogram of crystalline form 6 of compound A is shown in Figure 22.

[0199] Example 5 This example shows the characterization of crystalline forms of compound A by thermogravimetric measurement (TG) according to an embodiment of the present invention.

[0200] TG analysis was performed using a TA Instruments Discovery TGA 5500 instrument cooled using a TA Instruments low-temperature cooling system (RCS) 90 chiller. The balance of the instrument was calibrated using class M weights, and the temperature calibration was performed using alumel. Each sample was placed in a platinum sample pan, and the pan was loaded into the TG instrument. The pan was heated from ambient temperature to 350 °C at a rate of 10 °C per minute. It was controlled using TA Trios software v5.0.0.44608. The graph of thermogravimetric analysis (TGA) of crystalline form 1 of compound A free base is shown in Figure 4. The graph of thermogravimetric analysis (TGA) of crystalline compound A tosylate is shown in Figure 8. The graph of thermogravimetric analysis (TGA) of crystalline form 2 of compound A is shown in Figure 13. The graph of thermogravimetric analysis (TGA) of crystalline form 4 of compound A is shown in Figure 17. The graph of thermogravimetric analysis (TGA) of crystalline form 5 of compound A is shown in Figure 20. The graph of thermogravimetric analysis (TGA) of crystalline form 6 of compound A is shown in Figure 23.

[0201] Example 6 This example shows the crystalline forms of compound A according to an embodiment of the present invention 1Characterization by ¹H NMR and thermogravimetric measurement is shown.

[0202] 1 ¹H NMR spectra were acquired using TopSpin v3.2 software on a Bruker Avance 400 MHz spectrometer. Each sample was dissolved in DMSO-d6, and the resulting solution was transferred into a 5 mm NMR tube for subsequent data acquisition. The data collection parameters are shown in Table 7. The spectra were processed using the program MNova and referenced to the chemical shift of the residual protons in DMSO-d6. The 1 ¹H NMR spectrum of crystalline compound A free base is shown in Figure 5. The 1 ¹H NMR spectrum of crystalline compound A tosylate is shown in Figure 9.

[0203] [Table 7]

[0204] Example 7 This example shows the preparation of a crystalline form of compound A tosylate according to an embodiment of the present invention.

[0205] A heterogeneous mixture consisting of 19.7 mg (24.3 mmol) of compound A, 4.0 mg (21.0 mmol) of 4-toluenesulfonic acid monohydrate, and 1 mL of acetonitrile was stirred overnight at ambient temperature, then placed on a hot plate set at 40 °C and stirred for about 2 days. The mixture was cooled to ambient temperature and centrifuged. The liquid layer was decanted, and the remaining solid was dried in air to obtain a crystalline form of compound A tosylate.

[0206] Example 8 This example shows the characterization by X-ray powder diffraction (XRPD) of crystalline form 1 of compound A tosylate according to an embodiment of the present invention.

[0207] XRPD measurements were performed using one of three Rigaku Smart-Lab X-ray diffraction systems. Each was set to a Bragg-Brentano reflection configuration using a line-source X-ray beam. The Bragg-Brentano configuration was controlled by passive divergent and accepting slits, with the sample itself acting as a focusing element of the optical system. Powder samples were prepared in a low-background Si holder using light manual pressure to keep the sample surface flat and at the same height as the reference plane of the sample holder. The specifications of the diffractometer and data acquisition parameters for diffraction system 1 (file prefix RX1) are shown in Table 8. The specifications of the diffractometer and data acquisition parameters for diffraction system 2 (file prefix RX3) are shown in Table 9. The specifications of the diffractometer and data acquisition parameters for diffraction system 3 (file prefix RX4) are shown in Table 10. The X-ray powder diffraction pattern of crystalline compound A tosylate is shown in Figure 6. In the essentially pure material of crystalline compound A tosylate, a peak can be observed at a refraction angle of 2θ, as shown in Table 9.

[0208] [Table 8]

[0209] [Table 9]

[0210] [Table 10]

[0211] [Table 11-1]

[0212] [Table 11-2]

[0213] Example 9 This example shows the characterization of the crystal form of Compound A tosylate by dynamic vapor sorption (DVS) according to one embodiment of the present invention.

[0214] The DVS analysis was performed using a Q5000 sorption analyzer from TA Instruments. The instrument was calibrated using a standard brass weight and a sodium bromide standard for humidity. A portion of the sample was weighed into a metal-coated quartz pan. The analysis was carried out at 25 °C with a maximum equilibration time of 60 minutes in steps of 10% relative humidity (RH), from 5 to 95% RH (adsorption cycle) and 95 to 5% RH (desorption cycle). Data collection was performed using Advantage for Q Series version 5.5.23. The DVS graph of Compound A tosylate is shown in FIG. 10.

[0215] Example 10 This example shows an exemplary method for preparing and characterizing Crystal Form 2 of Compound A. Exemplary methods for preparing Crystal Form 2 of Compound A include: (1) equilibration of amorphous Compound A in isopropyl acetate or isopropyl alcohol; (2) reverse addition of a poor solvent using acetone and methyl tert-butyl ether (MTBE); and (3) equilibration of amorphous Compound A under a temperature cycle.

[0216] In the first method, about 30 mg of amorphous Compound A was dissolved in either isopropyl acetate or isopropyl alcohol and stirred at 25 °C for 1 week. The suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain Crystal Form 2.

[0217] In the second method, about 50 mg of amorphous Compound A was dissolved in a minimal amount of acetone at ambient temperature (e.g., about 20 - 25 °C) to form a clear solution. MTBE was added to this solution until a large amount of solid precipitated (about 1:4 (v / v) acetone / MTBE). The solid was recovered by centrifugation at 14,000 rpm through a 0.45 μm nylon membrane filter to obtain Crystal Form 2.

[0218] In the third method, approximately 30 mg of amorphous compound A was equilibrated in 0.1–0.3 mL of isopropyl acetate under a temperature cycle of 5°C–50°C at a heating / cooling rate of 0.1°C / min for 10 cycles. Equilibration was performed on a magnetic stirring plate using a stirring rod at a speed of 300–400 rpm. The resulting suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystalline form 2.

[0219] Crystal morphology 2 was characterized by XRPD, DSC, and TGA. According to XRPD, morphology 2 had a low degree of crystallinity (Figure 11). In the essentially pure material of crystal morphology 2, a peak can be observed at a refraction angle of 2θ, as shown in Table 12. By DSC, morphology 2 was characterized at 43.5°C TGA. onset At approximately 58 J / g enthalpy and 131.5°C T onset An enthalpy of approximately 6 J / g at 180.7°C, and a T of 180.7°C. onset It shows three desolvation / dehydration peaks with an enthalpy of approximately 3 J / g, followed by T in combination with decomposition. onset The melting peak persists at 205.7°C (Figure 12). TGA showed that morphology 2 exhibited a weight loss of approximately 5.1% at approximately 80°C and a weight loss of 2.3% between 80°C and 160°C (Figure 13).

[0220] [Table 12]

[0221] Example 11 This example illustrates a method for preparing and characterizing crystalline form 3 of compound A. Approximately 30 mg of amorphous compound A was dissolved in 0.1-0.3 mL of isopropyl alcohol and stirred at 25°C for one week. The suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystal morphology 3. Crystal morphology 3 was characterized by XRPD. According to XRPD, morphology 3 had a low degree of crystallinity (Figure 14). In the essentially pure material of crystal morphology 3, a peak can be observed at a refraction angle of 2θ, as shown in Table 13.

[0222] [Table 13]

[0223] Example 12 This example illustrates a method for preparing and characterizing crystalline form 4 of compound A. Exemplary methods for preparing crystalline form 4 of compound A include (1) equilibration of amorphous compound A in toluene or ethyl acetate / heptane; (2) equilibration of amorphous compound A under temperature cycling; and (3) equilibration of form 1 in ethyl acetate / heptane.

[0224] In the first method, approximately 30 mg of amorphous compound A was dissolved in 0.1-0.3 mL of toluene and stirred at 25°C for one week. The suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystalline form 4.

[0225] In the second method, approximately 30 mg of amorphous compound A was equilibrated in 0.1–0.3 mL of toluene under a temperature cycle of 5°C–50°C at a heating / cooling rate of 0.1°C / min for 10 cycles. Equilibration was performed on a magnetic stirring plate using a stirring rod at a speed of 300–400 rpm. The resulting suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystalline form 4.

[0226] In the third method, approximately 30 mg of compound A in crystalline form 1 was dissolved in 0.1-0.3 mL of ethyl acetate / heptane (1:1, v:v) and stirred at 25°C for one week. The suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystalline form 4.

[0227] Crystal morphology 4 was characterized by XRPD, DSC, and TGA. According to XRPD, morphology 4 had a low degree of crystallinity (Figure 15). In the essentially pure material of crystal morphology 4, a peak can be observed at a refraction angle of 2θ, as shown in Table 14. By DSC, morphology 4 was characterized at 30.9°C TGA. onset It showed a dehydration peak with an enthalpy of 63 J / g (Figure 16). There was no apparent melting after dehydration. By TGA, morphology 4 showed a weight loss of 4.6% at 80°C and a weight loss of 2.5% from 80°C to 160°C (Figure 17).

[0228] [Table 14]

[0229] Example 13 This example illustrates a method for preparing and characterizing crystalline form 5 of compound A, which has been identified as a hydrate.

[0230] Exemplary methods for preparing crystalline form 5 of compound A include (1) equilibration experiments using form 1, and (2) precipitation experiments by adding a poor solvent using form 1. In the first method, approximately 25 mg of compound A in crystalline form 1 was equilibrated on a magnetic stirring plate using a stirring rod at a speed of 300-400 rpm for 4 days at 50°C with either 0.1-0.3 mL of 1:2 (v:v) isopropyl alcohol / water or 1:2 (v:v) acetone / water. The resulting suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm to obtain crystalline form 5.

[0231] In the second method, approximately 50 mg of crystalline form 1 of compound A was dissolved in a minimal amount of isopropyl alcohol at ambient temperature (approximately 20-25°C). Water was slowly added to this solution until a large amount of solid precipitated (1:2 (v:v) isopropyl alcohol / water). The precipitate was collected by centrifugation at 14,000 rpm through a 0.45 μm nylon membrane filter to obtain crystalline form 5.

[0232] Crystal morphology 5 was characterized by XRPD, DSC, and TGA. According to XRPD, morphology 5 had a high degree of crystallinity (Figure 18). In the essentially pure material of crystal morphology 5, a peak can be observed at a refraction angle of 2θ, as shown in Table 15. By DSC, morphology 5 was characterized at 32.1°C TGA. onset A dehydration peak with an enthalpy of 48 J / g was observed at 81.2°C. onset The recrystallization peak at 172.5°C, and the T onset The melting peaks were observed with respect to recrystallization (Figure 19). TGA showed a weight loss of 4.8% at 150°C (Figure 20).

[0233] [Table 15-1]

[0234] [Table 15-2]

[0235] [Table 15-3]

[0236] Example 14 This example illustrates a method for preparing and characterizing the crystalline form 6 of compound A. Approximately 100 mg of compound A in crystalline form 1 was equilibrated in 0.5 mL of 1:1 (v:v) ethyl acetate / heptane at 25°C. Approximately 2 mg of seed crystals in form 4 were added to the mixture. The mixture was stirred at 25°C for 2 days, and the resulting solid was collected by vacuum filtration to obtain crystalline form 5.

[0237] Crystal morphology 6 was characterized by XRPD, DSC, and TGA. According to XRPD, morphology 6 had a low degree of crystallinity (Figure 21). In the essentially pure material of crystal morphology 6, a peak can be observed at a refraction angle of 2θ, as shown in Table 16. By DSC, morphology 6 was characterized at 53.3°C TGA. onsetA desolvation peak with an enthalpy of 44 J / g was observed, and at 166.4°C, T onset It showed an endothermic peak with an enthalpy of 4 J / g (Figure 22). By TGA, form 6 showed a weight loss of 6.7% at 150°C (Figure 23).

[0238] [Table 16]

[0239] Example 15 This example illustrates a method for preparing and characterizing the crystalline form 7 of compound A. Approximately 8 mg of compound A in crystalline form 1 was weighed and placed in an 8 mL glass vial. 4 mL of either FeSSIF-V1 (pH 5.0) or FaSSIF-V1 (pH 6.5) was added to this vial. The solution was stirred at 37°C and 400 rpm for 0.5 hours, 2 hours, and 24 hours, and then centrifuged at 37°C and 14,000 rpm for 5 minutes to obtain crystalline form 7. According to XRPD, form 7 had a low degree of crystallinity (Figure 24). In the essentially pure material of crystalline form 7, a peak can be observed at a refraction angle of 2θ, as shown in Table 17.

[0240] [Table 17]

[0241] Example 16 This embodiment illustrates an exemplary method for preparing and characterizing crystalline form 8 of compound A. Crystalline form 8 was obtained by heating crystalline form 1 of compound A to 120°C at 0% RH. When RH was increased to 40%, crystalline form 8 reverted to crystalline form 1. According to XRPD, form 8 had a low degree of crystallinity (Figure 25). In the essentially pure material of crystalline form 8, a peak can be observed at a refraction angle of 2θ, as shown in Table 18. Figure 26 shows an overlay of XRPDs of crystalline forms 1-8 of compound A.

[0242] [Table 18]

[0243] Other Embodiments While the present invention is described in relation to its specific embodiments, it should be understood that the invention is subject to further modifications, and this application is intended to encompass any variations, uses, or adaptations of the invention, including any deviations from this disclosure that, generally in accordance with the principles of the invention, are included in known or customary practices of the relevant art and can be applied to the essential features described herein.

[0244] All publications, patents, and patent applications are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated as being incorporated herein by reference in whole.

Claims

1. Crystalline solid form of compound A: 【Chemistry 1】 or its solvate.

2. The crystalline solid form according to claim 1, wherein compound A or its solvate is selected from form 1, form 2, form 3, form 4, form 5, form 6, form 7, or form 8.

3. The crystalline solid form according to claim 1 or 2, wherein compound A or the solvate thereof is form 1.

4. The crystalline solid form according to claim 1 or 2, wherein compound A or the solvate thereof has form 5.

5. The crystalline hydrate of compound A. 【Chemistry 2】

6. The crystalline solid form or solvate thereof according to claim 3, having at least one peak at diffraction angles 2θ (°) of 5.2 ± 0.5, 8.4 ± 0.5, or 10.0 ± 0.5 when measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction.

7. Crystallographic form of compound A: 5 【Transformation 3】 or a solvate thereof, which, when measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction, has at least one peak at diffraction angles 2θ (°) of 14.3 ± 0.5, 16.8 ± 0.5, or 20.5 ± 0.5, is compound A or the solvate thereof.

8. The crystalline solid form of the tosylate salt of compound A, or its solvate. 【Chemistry 4】

9. The crystalline solid form or solvate thereof according to claim 8, having at least one peak at diffraction angles 2θ (°) of 5.9 ± 0.5, 9.4 ± 0.5, or 9.8 ± 0.5 when measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction.

10. A pharmaceutical composition comprising a crystalline form of compound A according to any one of claims 1 to 9, or a solvate thereof, and a pharmaceutically acceptable carrier or excipient.

11. Crystal form of compound A, 1. 【Transformation 5】 or a method for producing a solvate thereof, the method comprising: preparing a slurry of compound A in a suitable solvent or buffer to precipitate the crystalline solid of form 1; isolating the crystalline solid of form 1 from the slurry; and drying the crystalline solid of form 1.

12. Crystal form of compound A, 5 【Transformation 6】 or a method for producing a solvate thereof, the method comprising: preparing a slurry of crystalline form 1 of compound A in a suitable solvent to precipitate the crystalline solid of form 5; isolating the crystalline solid of form 5 from the slurry; and drying the crystalline solid of form 5.

13. Crystal form of compound A, 5 【Transformation 7】 or a method for producing a solvate thereof, the method comprising: preparing a mixture of crystalline form 1 of compound A in isopropyl alcohol; precipitating a crystalline solid of form 5 by adding water; isolating the crystalline solid of form 5 from the slurry; and drying the crystalline solid of form 5.

14. Crystalline form of the tosylate salt of compound A, 【Transformation 8】 or a method for producing a solvate thereof, the method comprising: preparing a mixture of compound A and 4-toluenesulfonic acid in a suitable solvent or buffer in order to precipitate a crystalline solid of the tosylate of compound A; isolating the crystalline solid of the tosylate of compound A from the mixture; and drying the crystalline solid of the tosylate of compound A.

15. A method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of compound A in crystalline form or a solvate thereof, or the pharmaceutical composition described in claim 10, according to any one of claims 1 to 9.

16. A method for treating a Ras protein-related disease in a subject requiring treatment for the Ras protein-related disease, the method comprising administering to the subject a therapeutically effective amount of compound A in crystalline form or a solvate thereof, or the pharmaceutical composition described in claim 10, according to any one of claims 1 to 9.

17. A method for inhibiting intracellular Ras protein, wherein the method comprises contacting the cells with an effective amount of compound A in crystalline form or cultured form thereof as described in any one of claims 1 to 9, or with the pharmaceutical composition as described in claim 10.

Images