Crystal morphology of RAS inhibitors

Abstract

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

Description

[Technical Field]

[0001] This invention relates to the crystalline form of RAS inhibitors. [Background technology]

[0002] The vast majority of small molecule drugs act by binding to functionally important pockets on target proteins, thereby modulating the activity of those proteins. For example, cholesterol-lowering drugs known as statins bind to the enzymatic active site of HMG-CoA reductase, thus preventing the enzyme from engaging with its substrate. The fact that many such drug / target interaction pairs are known may lead some to believe that, given a reasonable amount of time, effort, and resources, small molecule modifiers 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. Non-patent document 1. The other 90% are currently considered refractory or refractory to the small molecule drug discovery described above. Such targets are commonly referred to as "un-drug-worthy." These un-drug-worthy targets represent a vast and large untapped stock of clinically important human proteins. Therefore, there is considerable interest in discovering novel molecular modalities capable of modulating the function of such un-drug-worthy targets.

[0003] The Ras proteins (K-Ras, H-Ras, and N-Ras) are well-established in the literature as playing essential roles in various human cancers and therefore being suitable targets for anticancer therapy. In fact, mutations in the Ras protein account for approximately 30% of all human cancers in the United States, many of which are fatal. Dysregulation of the Ras protein due to activating mutations, overexpression, or upstream activation is common in human tumors, and activating mutations in Ras are frequently found in human cancers. For example, an activating mutation at codon 12 in the Ras protein functions by inhibiting both the GTPase-activated protein (GAP)-dependent and intrinsic hydrolysis rates of GTP, significantly distorting the population of Ras mutant proteins into an "on" (GTP-bound) state (Ras(ON)), leading to oncogenic MAPK signaling. In particular, Ras exhibits picomolar affinity for GTP, allowing Ras to be activated even in the presence of low concentrations of this nucleotide. Mutations in codons 13 (e.g., G13C) and 61 (e.g., Q61K) of Ras are also involved in oncogenic activity in some cancers.

[0004] Despite extensive drug discovery efforts against Ras over the past several decades, only two drugs targeting Ras, namely K-Ras, have emerged. G12C Sotrasib and adagrav, which target Ras mutations, are approved in the United States. Further efforts are needed to identify additional drugs for cancers caused by various Ras mutations. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Bojadzic and Buchwald, Curr Top Med Chem 18:674-699 (2019) [Overview of the project] [Means for solving the problem]

[0006] The present invention features crystalline forms of compounds useful for treating diseases or conditions (e.g., cancer, Ras protein-related disorders). In some embodiments, this disclosure describes the crystalline form of compound A.

[0007] [ka]

[0008] In some embodiments, the crystalline form of compound A or its solvate is selected from Form 1, Form 2, Form 3, or Form 4. In some embodiments, the crystalline form of compound A or its solvate is Form 1.

[0009] In some embodiments, crystalline form 1 of compound A or its solvate has at least one peak at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, or 5.1±0.5, as measured by or calculated from X-ray diffraction by irradiation with Cu Kα X-rays. In some embodiments, crystalline form 1 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, and 5.1±0.5, as measured by or calculated from X-ray diffraction by irradiation with Cu Kα X-rays. In some embodiments, crystalline form 1 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 7.5±0.5, 9.4±0.5, and 9.8±0.5, as measured by or calculated from X-ray diffraction by irradiation with Cu Kα X-rays. In some embodiments, crystalline form 1 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, 5.1±0.5, 7.5±0.5, 9.4±0.5, and 9.8±0.5, as measured by X-ray diffraction induced by Cu Kα X-ray irradiation 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 10.3±0.5, 10.7±0.5, and 11.2±0.5, as measured by X-ray diffraction induced by Cu Kα X-ray irradiation 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 4.4±0.5, 4.6±0.5, 5.1±0.5, 7.5±0.5, 9.4±0.5, 9.8±0.5, 10.3±0.5, 10.7±0.5, and 11.2±0.5, as measured by or calculated from X-ray diffraction by Cu Kα X-ray irradiation. In some embodiments, crystalline form 1 of compound A or its solvate has an X-ray powder diffractogram as shown in Figure 1.

[0010] In some embodiments, the crystalline form 1 of compound A is a hydrate. In some embodiments, the crystalline form 1 of compound A is a mixed isopropyl ether, ethanol, and aqueous solvate. In some embodiments, the crystalline form 1 of compound A is a mixed diethyl ether and aqueous solvate. In some embodiments, the crystalline form 1 of compound A has parameters a=40.5965Å, b=16.0423Å, c=19.4198Å, and V=12,647.4Å. 3 A mixed isopropyl ether, ethanol, and aqueous solvate, further characterized by a unit cell having the following parameters: a=40.813 Å, b=16.079 Å, c=19.093 Å, and V=12,529 Å. 3 A mixed diethyl ether and aqueous solvate, further characterized by a unit cell having [a specific characteristic].

[0011] In some embodiments, crystalline form 1 of compound A or its solvate exhibits an endothermic onset at 163.4°C ± 0.5°C in differential scanning calorimetry (DSC) profiles. In some embodiments, crystalline form 1 of compound A or its solvate has a DSC thermogram as shown in Figure 7. In some embodiments, crystalline form 1 of compound A or its solvate exhibits a weight loss of 0.4% ± 0.5 (w / w) at ambient ~150.0°C ± 0.5°C or 0.5% ± 0.5 (w / w) at ambient ~200.0°C ± 0.5°C in thermogravimetric analysis (TGA) profiles. In some embodiments, crystalline form 1 of compound A or its solvate has a TGA graph as shown in Figure 7.

[0012] In one embodiment, the present invention relates to a mixture of crystalline forms 1 and 2 of compound A:

[0013] [ka]

[0014] The mixture or its solvate is characterized by having at least one peak at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, or 4.8±0.5, as measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, the mixture of crystalline forms 1 and 2 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, and 4.8±0.5, as measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, the mixture of crystalline forms 1 and 2 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 5.1±0.5, 6.1±0.5, and 7.4±0.5, as measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, 4.8±0.5, 5.1±0.5, 6.1±0.5, and 7.4±0.5, as measured by X-ray diffraction irradiated with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 8.0±0.5, 9.4±0.5, and 10.3±0.5, as measured by X-ray diffraction irradiated with Cu Kα X-rays or calculated from X-ray diffraction. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, 4.8±0.5, 5.1±0.5, 6.1±0.5, 7.4±0.5, 8.0±0.5, 9.4±0.5, and 10.3±0.5, as measured by or calculated from X-ray diffraction by Cu Kα X-ray irradiation. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has an X-ray powder diffractogram as shown in Figure 2.

[0015] In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate exhibits endothermic peaks at 69.1°C ± 0.5 and 171.4°C ± 0.5 in differential scanning calorimetry (DSC) profiles. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has a DSC thermogram as shown in Figure 8. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate exhibits a weight loss of 0.71% ± 0.5 (w / w) at ambient ~150.0°C ± 0.5 or 0.74% ± 0.5 (w / w) in thermogravimetric analysis (TGA) profiles. In some embodiments, a mixture of crystalline forms 1 and 2 of compound A or its solvate has a TGA graph as shown in Figure 8.

[0016] In some embodiments, the present invention is characterized by a pharmaceutical composition comprising 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 crystalline form 1 of compound A or a mixture of crystalline forms 1 and 2:

[0017] [ka]

[0018] or a method for preparing a solvate thereof, characterized by the above method comprising: dissolving compound A in a suitable solvent; precipitating the crystalline form(s) of compound A by adding a suitable antisolvent; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A. In some embodiments, the suitable solvent is isopropyl ether and the suitable antisolvent is ethanol. In some embodiments, the suitable solvent is a mixture of an organic acid and diethyl ether. In some embodiments, the suitable solvent is ethyl acetate and the suitable antisolvent is hexane.

[0019] In one embodiment, the present invention relates to crystalline form 1 of compound A or a mixture of crystalline forms 1 and 2:

[0020] [ka]

[0021] or a method for preparing a solvate thereof, characterized by the method comprising: dissolving compound A in a suitable solvent; precipitating the crystalline form(s) of compound A by evaporation of the suitable solvent; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A. In some embodiments, the suitable solvent is a mixture of diethyl ether and hexane.

[0022] In one embodiment, the present invention relates to crystalline form 1 of compound A or a mixture of crystalline forms 1 and 2:

[0023] [ka]

[0024] or a method for preparing a solvate thereof, characterized by the above method comprising: dissolving compound A in a suitable solvent; precipitating the crystalline form(s) of compound A under ambient conditions; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A. In some embodiments, the suitable solvent is diethyl ether, or a mixture of ethyl acetate and isopropyl ether.

[0025] In some embodiments, the present invention is characterized by a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of crystalline form 1 or a mixture of crystalline forms 1 and 2, or a solvate thereof, or a pharmaceutical composition thereof. In some embodiments, the cancer comprises a Ras mutation. In some embodiments, the Ras mutation is G12C. 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.

[0026] In some embodiments, the present invention is characterized by a method for treating Ras protein-related disorders in subjects requiring treatment for Ras protein-related disorders, the method comprising administering to the subject a therapeutically effective amount of crystalline form 1 of compound A, a mixture of crystalline forms 1 and 2 of compound A, a solvate thereof, or a pharmaceutical composition.

[0027] In some embodiments, the present invention is characterized by a method for inhibiting intracellular Ras proteins, the method comprising administering to a subject an effective amount of crystalline form 1 or a mixture of crystalline forms 1 and 2 of compound A, or a solvate thereof, or a pharmaceutical composition thereof. In some embodiments, more than one Ras protein is 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 colon cancer cells. In some embodiments, the Ras protein is KRAS.

[0028] In some embodiments, the method further includes 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 RAS MULTI It is an inhibitor. In some embodiments, the second Ras inhibitor is RAS MULTI It is an (ON) inhibitor. In some embodiments, RAS MULTI (ON) inhibitors include:

[0029] [ka]

[0030] or a pharmaceutically acceptable salt thereof. Any limitations discussed in relation to one embodiment of the present invention are specifically intended to be applicable to any other embodiments of the present invention. Furthermore, any compound or composition of the present invention may be used in any method of the present invention, and any method of the present invention may be used to produce or utilize any compound or composition of the present invention.

[0031] Definitions and Chemical Terms In this application, unless otherwise made clear from the context, (i) the term “a” means “one or more”; (ii) the term “or” is used to mean “and / or” unless it is explicitly indicated that it refers only to substitutes or that the substitutes are mutually exclusive, however this disclosure supports the definitions of substitutes only and “and / or”; (iii) the terms “comprising” and “including” are understood to encompass itemized components or steps, whether presented alone or together with one or more additional components or steps; and (iv) if a scope is provided, it includes endpoints.

[0032] 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.

[0033] As used herein, the term “adjacent” in the context of describing adjacent atoms refers to divalent atoms that are directly bonded by a covalent bond. "Crystal form of compound" and similar terms as used herein 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 explicitly stated herein.

[0034] The term "wild-type" refers to an entity possessing a structure or activity found in nature in a "normal" state or context (as opposed to mutants, pathological, or degenerated forms). Those skilled in the art will understand that wild-type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0035] 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 can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials, such as by resolution of racemic mixtures or stereoselective synthesis, are known in the art. Many geometric isomers, including olefins and C=N double bonds, may also exist among the compounds described herein, and all such stable isomers are intended in this disclosure. The cis and trans geometric isomers of the compounds of this disclosure are described and can be isolated as mixtures of isomers or as separated isomers.

[0036] 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 a proton. In certain embodiments, the tautomerized form may be a prototropic tautomer, which is the 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 a proton 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-isoindole, and 1H- and 2H-pyrazole. In some embodiments, the tautomer morphs are in equilibrium or can be sterically fixed to one morph by appropriate substitution. In certain embodiments, the tautomer morphs arise from acetal interconversion.

[0037] Details of one or more embodiments of the present invention are described below. Other features, purposes, and advantages of the present invention will become apparent from the description and claims. [Brief explanation of the drawing]

[0038] [Figure 1] This is an exemplary X-ray powder diffractogram of crystalline form 1 of compound A as a mixed ethanol and isopropyl ether solvate. [Figure 2] This is an exemplary X-ray powder diffractogram of a mixture of crystalline forms 1 and 2 of compound A. [Figure 3] This is an overlay of exemplary X-ray powder diffractograms of pure crystalline form 1 of compound A, and a mixture of crystalline forms 1 and 2 of compound A. [Figure 4]This is an exemplary X-ray powder diffractogram overlay showing the formation of a mixture of crystalline forms 1 and 2 of compound A, starting from pure crystalline form 1 of compound A over time. [Figure 5] This is an exemplary X-ray crystal structure of crystalline form 1 of compound A as a mixed ethanol, isopropyl ether, and aqueous solvate (with chiral units shown). [Figure 6] This is an exemplary X-ray crystal structure of crystalline form 1 of compound A as a mixed diethyl ether and aqueous solvate (with chiral units shown). [Figure 7] This is an exemplary differential scanning calorimetry (DSC) thermogram and an exemplary thermogravimetric analysis (TGA) overlay of crystalline form 1 of compound A. [Figure 8] Exemplary DSC thermogram and exemplary TGA overlay of a mixture of crystalline forms 1 and 2 of compound A. [Figure 9] This is an exemplary X-ray powder diffractogram of crystalline form 3 of compound A. [Figure 10] This is an exemplary DSC thermogram of crystalline form 3 of compound A. [Figure 11] This is an exemplary TGA of crystalline form 3 of compound A. [Figure 12] This is an exemplary X-ray powder diffractogram of crystalline form 4 of compound A. [Figure 13] This is an exemplary X-ray powder diffractogram of crystalline form 4 of compound A after two weeks of storage at room temperature. [Figure 14] This is an exemplary DSC thermogram and exemplary TGA overlay of crystalline form 4 of compound A. [Modes for carrying out the invention]

[0039] compound In general, the present invention provides a crystalline form of formula I. The compound of formula I, hereafter referred to as compound A, has the following structure.

[0040] [ka]

[0041] The crystalline form of compound A may be, for example, crystalline form 1, crystalline form 2, or a mixture of forms 1 and 2. Hereafter, the crystalline forms of compound A will be identified by their unique XRPD patterns; that is, hereafter, crystalline form 1 of compound A will be referred to synonymously with form 1.

[0042] As described in the examples, Form 1 or its solvate may have one or more peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, 5.1±0.5, 7.5±0.5, 9.4±0.5, 9.8±0.5, 10.3±0.5, 10.7±0.5, and 11.2±0.5, as measured by X-ray diffraction by irradiation with Cu Kα X-rays or calculated from X-ray diffraction. Form 1 as a mixed ethanol and isopropyl ether solvate may have the X-ray powder diffractogram shown in Figure 1.

[0043] A mixture of forms 1 and 2 of compound A, or its solvate, may have one or more peaks at diffraction angles 2θ(°) of 4.4±0.5, 4.6±0.5, 4.8±0.5, 5.1±0.5, 6.1±0.5, 7.4±0.5, 8.0±0.5, 9.4±0.5, and 10.3±0.5, as measured by X-ray diffraction irradiated with Cu Kα X-rays or calculated from X-ray diffraction. A mixture of crystalline forms 1 and 2 of compound A, or its solvate, may have the X-ray powder diffractogram shown in Figure 2.

[0044] Form 1 may have a crystalline structure as a mixed ethanol, isopropyl ether, and aqueous solvate as shown in Figure 5. Form 1 may have a crystalline structure as a mixed diethyl ether and aqueous solvate as shown in Figure 6.

[0045] Form 1 or its solvate may exhibit an endothermic onset at 163.4°C ± 0.5°C as determined by differential scanning calorimetry (see Figure 7). Form 1 or its solvate may exhibit a weight loss of 0.37% ± 0.5 (w / w) at ambient ~150.0°C ± 0.5°C and 0.49% ± 0.5 (w / w) at ambient ~200.0°C ± 0.5°C in the thermogravimetric analysis profile (see Figure 7). A mixture of Forms 1 and 2, or its solvate, may exhibit endothermic peaks at 69.1°C ± 0.5°C and 171.38°C ± 0.5°C as determined by differential scanning calorimetry (see Figure 8). Mixtures of Forms 1 and 2 or their solvates may exhibit a weight loss of 0.71% ± 0.5 (w / w) at ambient temperature ~150.0°C ± 0.5 and 0.74% ± 0.5 (w / w) at ambient temperature ~200.0°C ± 0.5 in the thermogravimetric analysis profile (see Figure 8).

[0046] Furthermore, a method for treating cancer in subjects requiring cancer treatment is 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 K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C. Other Ras mutations are described herein.

[0047] A method is provided for treating Ras protein-related disorders in subjects requiring treatment of such disorders, the method comprising administering a therapeutically effective amount of the crystalline compound of the present invention to the subject.

[0048] Furthermore, a method for inhibiting intracellular Ras proteins is provided, which comprises contacting cells with an effective amount of the crystalline compound of the present invention. For example, the Ras proteins are K-Ras G12C, K-Ras G13C, H-Ras G12C, H-Ras G13C, N-Ras G12C, or N-Ras G13C. 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.

[0049] 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.

[0050] Synthesis method The compounds described herein may be prepared from commercially available starting materials or synthesized using known organic, inorganic, or enzymatic processes.

[0051] The compounds of the present invention can be prepared by several methods well known to those skilled in the art of organic synthesis. Exemplary synthesis of the compounds of the present invention is disclosed in WO2021 / 091982, which is incorporated herein by reference.

[0052] Pharmaceutical composition and method of use The crystalline form of the compound relating to the present invention is a Ras inhibitor and is useful in the treatment of cancer. Accordingly, one embodiment of the present invention provides a pharmaceutical composition containing the crystalline form of the compound 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.

[0053] As used herein, the term “pharmaceutical composition” refers to a compound or crystalline form, such as the crystalline compound of the present invention, formulated together with pharmaceutically acceptable excipients. In some embodiments, the crystalline form(s) of the compound is present in the pharmaceutical composition at a unit dose appropriate for administration in a therapeutic regimen, exhibiting a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including oral administration, e.g., oral tablets (aqueous or nonaqueous solutions or suspensions), tablets, e.g., targeted for oral, sublingual, and intracellular absorption, boluses, powders, granules, pastes for application to the tongue, e.g., sterile solutions or suspensions, or parenteral administration as a sustained-release formulation, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., as a cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; e.g., as a pessary, cream, or foam adapted for vaginal or rectal administration, sublingual administration, intraocular administration, transdermal administration, or transnasal administration, intrapulmonary administration, and administration to other mucosal surfaces.

[0054] 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, fluidizers (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, or hydration 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 useful excipients and materials.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.

[0055] 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.

[0056] As used herein, the term “dosage form” refers to a physically distinct unit of a compound (e.g., the 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 according to a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., using a therapeutic dosing regimen). Those skilled in the art will understand that the total amount of a therapeutic composition or compound to be administered to a particular subject may be determined by one or more attending physicians and may involve administration in multiple dosage forms.

[0057] As used herein, the term “dosage regimen” means a set of individual unit doses (typically more than one) administered to a subject, typically separated by periods. In some embodiments, a given therapeutic compound (e.g., the crystalline compounds of the present invention) has a recommended dosage regimen which may consist of one or more doses. In some embodiments, the dosage regimen comprises multiple doses, each separated from the others by periods of equal length, and in some embodiments, the dosage regimen comprises multiple doses and at least two different periods separating the individual doses. In some embodiments, all doses within the dosage regimen are the same unit dose. 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 additional 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 additional doses at a second dose the same as the first dose. In some embodiments, the administration regimen correlates with a desired or beneficial outcome when administered across a relevant population (i.e., it is a therapeutic administration regimen).

[0058] A "treatment regimen" refers to a regimen of administration that, when administered across a relevant population, correlates with a desired or beneficial treatment outcome. The term “treatment” (similarly, “to treat” or “to treat”), in its broadest sense, refers to any administration of a substance (e.g., the crystalline compounds of the present invention) that partially or completely alleviates, improves, mitigates, inhibits, delays the onset of, reduces the severity of, or reduces the incidence 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 early signs of the disease, disorder, or condition. Alternatively, or in addition, in some embodiments, 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 treatment of a subject diagnosed with a relevant disease, disorder, or condition. In some embodiments, treatment may be treatment of a subject known to have one or more susceptibility factors that are statistically correlated with an increased risk of progression of the relevant disease, disorder, or condition.

[0059] The term “therapeutic dose” means an amount sufficient to treat a disease, disorder, or condition when administered to a population suffering from or susceptible to such a disease, disorder, or condition, according to a therapeutic administration regimen. In some embodiments, a therapeutic dose is one that reduces the incidence or severity of one or more symptoms of a disease, disorder, or condition, or delays their onset. Those skilled in the art will understand that the term “therapeutic dose” does not actually require that the success of the treatment be achieved in a particular individual. Rather, a therapeutic dose may be an amount that, when administered to a subject in need of such treatment, provides a particular desired pharmacological response in a significant number of subjects. It is specifically understood that a particular subject may actually be “refractory” to a “therapeutic dose.” In some embodiments, a reference to a therapeutic dose may refer to an amount 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, a therapeutic 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.

[0060] For use as a treatment of a target, the crystalline form of the compounds of the present invention can be formulated as a pharmaceutical or veterinary composition. Depending on the target to be treated, the mode of administration, and the type of treatment desired, e.g., prevention, prophylaxis, or therapeutic approach, the compounds are formulated in a manner consistent with these parameters. Outlines of such techniques are incorporated herein by reference, Remington: The Science and Practice of Pharmacy, 21 st This 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.

[0061] Each composition can be prepared according to conventional mixing, granulation, or coating methods, and the pharmaceutical compositions may contain, by weight or volume, about 0.1% to about 99%, about 5% to about 90%, or about 1% to about 20% of the crystalline compounds of the present invention. In some embodiments, the crystalline forms of the compounds described herein may be present in an amount totaling 1 to 95% by weight of the total weight of a composition such as a pharmaceutical composition.

[0062] The composition may be provided in dosage forms suitable for intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, cutaneous, subcutaneous, topical, transdermal, sublingual, transnasal, intravaginal, intravesical, intraurethral, ​​intrathecal, epidural, transaural, or intraocular administration, or by injection, inhalation, or administration by direct contact with the nasal, genitourinary, genital, 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.

[0063] As used herein, the term “administration” means the administration of a composition (e.g., a crystalline form of compound A, or a preparation comprising a crystalline form of compound A as described herein) to a subject or system. Administration to an animal subject (e.g., a human) may be by any suitable route. For example, in some embodiments, administration may be by trachea (including by intratracheal instillation), oral cavity, intestinal, intradermal, intra-arterial, intradermal, gastric, intramedullary, intramuscular, intranasal, intraperitoneal, intramedullary, intravenous, intraventricular, mucosa, intranasal cavity, oral, rectal, subcutaneous, sublingual, topical, trachea (including intratracheal instillation), transdermal, vaginal, or intravitreal administration.

[0064] Formulations may be prepared in a form suitable for systemic administration or topical or local administration. Systemic formulations may include those designed for injection (e.g., intramuscular, intravenous, or subcutaneous injection) or may be prepared for transdermal, transmucosal, or oral administration. Formulations generally include diluents, and optionally adjuvants, buffers, and preservatives. The crystalline form of the compound may be administered as a liposome composition or as a microemulsion.

[0065] For injection, formulations can be prepared in conventional forms, such as solutions or suspensions, or as solid forms suitable for solutions or suspensions in liquids before injection, or as emulsions. Suitable excipients include, for example, water, physiological saline, dextrose, and glycerol. Such compositions may also contain amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, such as pH buffers such as sodium acetate and sorbitan monolaurate.

[0066] Various sustained-release systems for drugs have also been devised. See, for example, U.S. Patent No. 5,624,677. Systemic administration may also include relatively non-invasive methods such as the use of suppositories, transdermal patches, transmucosal delivery, and intranasal administration. Oral administration is also suitable for the compounds of the present invention. Preferred forms include syrups, capsules, and tablets, as understood in the art.

[0067] Each crystalline form of the compounds described herein can be formulated in various ways known in the art. For example, the first and second agents of combination therapy can be formulated together or separately. Other modalities of combination therapy are described herein.

[0068] Individually or separately formulated drugs can be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing, for example, two pills, pills and powder, suppositories and liquids in vials, two topical creams, etc. Kits may include any 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, inhalers, etc. In addition, unit dose kits may include instructions for the preparation or administration of the composition. Kits may be manufactured as single-use unit doses for one subject, as multiple-use units for a specific subject (at a constant dose, or where the potency of individual compounds may change as the treatment progresses), or kits may contain multiple doses suitable for administration to multiple subjects ("bulk packaged"). The components of a kit may be assembled into cartons, blister packs, bottles, tubes, etc.

[0069] Preparations for oral use include tablets containing the active ingredient(s) in a mixture with non-toxic, pharmaceutically acceptable 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, fluidizers, 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, and buffering agents.

[0070] Two or more compounds may be mixed together or distributed in a tablet, capsule, or other vehicle. In one example, the first compound may be contained inside the tablet, and the second compound may be on the outside such that a substantial portion of the second compound is released before the release of the first crystalline compound.

[0071] Formulations for oral use may be provided as chewable tablets, or as rigid 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 a water or oily medium, such as peanut oil, liquid paraffin, or olive oil. Powders, granules, and pellets may be prepared using the above-mentioned components, for example, in a conventional manner using a mixer, fluidized bed apparatus, or spray dryer, along with tablets and capsules.

[0072] Dissolution or diffusion-controlled release can be achieved by appropriate coating of tablets, capsules, pellets, or granular formulations 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 controlled-release matrix formulations, the matrix material may also include, for example, hydrated methylcellulose, carnauba wax and stearyl alcohol, Carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, or halogenated fluorocarbons.

[0073] Liquid forms in which the crystalline forms of the compounds and compositions of the present invention can be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions containing edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0074] Generally, when administered to humans, the oral dosage 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 dosage may be, for example, about 0.001 mg to about 2000 mg per day, about 1 mg to about 1000 mg per day, about 5 mg to about 500 mg per day, about 100 mg to about 1500 mg per day, about 500 mg to about 1500 mg per day, about 500 mg to about 2000 mg per day, or any range that can be derived within that range.

[0075] In some embodiments, the pharmaceutical composition may further comprise additional compounds having antiproliferative activity. Depending on the mode of administration, the compounds or pharmaceutically acceptable salts thereof are formulated into a composition suitable for easy delivery. Each compound or pharmaceutically acceptable salt thereof in a combination therapy may be formulated in a variety of ways known in the art. For example, the first and second agents of a combination therapy may be formulated together or separately. Preferably, the first and second agents are formulated together for simultaneous or near-simultaneous administration.

[0076] 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. Specific combinations of treatments (therapeutic agents or procedures) for use in combination regimens take into account the suitability of the desired therapeutic agents or procedures and the desired therapeutic effect to be achieved. It will also be understood that the treatments used may achieve the desired effect against the same disorder or different effects (e.g., control of any adverse effects).

[0077] As described herein, the administration of each drug in combination therapy may be independently once to four times daily for a period of one day to one year, or even for the lifetime of the subject. Chronic long-term administration may be indicated.

[0078] How to use In some embodiments, the present invention discloses methods for treating diseases or disorders characterized by abnormal Ras activity resulting from Ras variants. In some embodiments, the disease or disorder is cancer.

[0079] Accordingly, a method for treating cancer in subjects requiring treatment for cancer is also provided, which comprises administering to a subject a therapeutically effective amount of the compound of the present invention in crystalline form, or a pharmaceutical composition containing such a crystalline form of the compound or a salt. 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 subjects requiring treatment for Ras protein-related disorders is also provided, which comprises administering to a subject a therapeutically effective amount of the compound of the present invention in crystalline form, or a pharmaceutical composition containing such a crystalline compound or a salt.

[0080] In some embodiments, the crystalline forms of the compounds of the present invention, pharmaceutical compositions containing such crystalline forms, and methods provided herein may be used to treat a wide variety of cancers, including tumors of the lung, prostate, breast, brain, skin, cervical cancer, and testicular cancer. More specifically, cancers that can be treated by the compounds or salts thereof of the present invention, pharmaceutical compositions containing such compounds or salts, and methods include, but are not limited to, astrocytic, mammary, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocyte, laryngopharyngeal, lung, oral cavity, ovarian, prostate, and thyroid cancer and sarcoma. Other cancers include, for example: The heart, for example, sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyomas, fibromas, lipomas, and teratomas, Lung cancer, for example, bronchogenic carcinoma (squamous cell carcinoma, anaplastic small cell carcinoma, anaplastic large cell carcinoma, adenocarcinoma), alveolar (bronchial) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondrotoxic hamartoma, mesothelioma, The gastrointestinal tract, for example, the esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), the stomach (carcinoma, lymphoma, leiomyosarcoma), the pancreas (ductal adenocarcinoma, islet cell tumor, glucagon-producing tumor, gastrin-producing tumor, carcinoid tumor, VIP-producing tumor), the small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), the large intestine (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma), The urogenital system, for example, the kidneys (adenocarcinoma, Wilms' tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testes (seminocarcinoma, teratoma, embryonic carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenoid tumor, lipoma), Liver, for example, liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, Biliary tract, for example, gallbladder cancer, ampulla cancer, bile duct cancer, Bone, such as osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteocartilaginous exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor, Nervous system, such as skull (osteoma, hemangioma, granuloma, xanthoma, Paget's disease), meninges (meningioma, meningosarcoma, gliosis), brain (astrocytoma, medulloblastoma, glioma, epithelioma, germ cell tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), spinal nerve fibroma, neurofibromatosis type I, meningioma, glioma, sarcoma), Gynecology, such as 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 germ 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 (carcinoma), Hematological, such as blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia), myeloproliferative disorders (e.g., myelofibrosis and myeloproliferative neoplasia), multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin lymphoma (malignant lymphoma), Skin, such as malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis, and Adrenal gland, such as neuroblastoma.

[0081] 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 that includes Ras WT (e.g., K-Ras WT , H-Ras WT , or N-Ras WT ). In some embodiments, the Ras protein is Ras amplification (e.g., K-Ras ampTherefore, in some embodiments, the crystalline compound of the present invention is Ras amp (K-Ras amp H-Ras amp , or N-Ras amp It is used in methods for treating patients with cancer including ). In some embodiments, the cancer includes Ras mutations such as the Ras mutations described herein. In some embodiments, the mutations are selected from the following: (a) The following K-Ras mutants: 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, as well as (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 crystalline compounds of the present invention inhibit one 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 compounds of the present invention inhibit one or more additional Ras mutants (e.g., K-, H-, or N-Ras WT In addition to K-Ras G12C or G13C, or a combination thereof, Ras WTIt inhibits. In some embodiments, the crystalline compounds of the present invention have one or more additional Ras mutations (e.g., K-, H-, or N-Ras). amp In addition to G12C or G13C, or a combination thereof, Ras amp To inhibit.

[0082] Methods for detecting Ras mutations are known in the art. Such means include, but are not limited to, direct sequencing and the use of highly sensitive diagnostic assays (using CE-IVD marks), such as those described in Domagala, et al., Pol J Pathol 3:145-164 (2012), which are incorporated herein by reference in their entirety, including TheraScreen PCR, AmoyDx, PNAClamp, RealQuality, EntroGen, LightMix, StripAssay, Hybcell plexA, Devyser, Surveyor, Cobas, and TheraScreen Pyro. See also, for example, WO2020 / 106640.

[0083] In some embodiments, the cancer is non-small cell lung cancer, and the Ras mutation includes a K-Ras mutation such as K-Ras G12C. In some embodiments, the cancer is colorectal cancer, and the Ras mutation includes a K-Ras mutation such as K-Ras G12C.

[0084] 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 Includes mutations. In some embodiments, the cancer is characterized by the K-Ras G13C mutation and STK11 LOF, comprising KEAP1, EPHA5, or NF1 mutations. In some embodiments, the cancer is colorectal cancer and comprises the K-Ras G12C mutation. In some embodiments, the cancer is endometrial cancer, ovarian cancer, cholangiocarcinoma, or mucinous appendiceal cancer and comprises the K-Ras G12C mutation. In some embodiments, the cancer is gastric cancer and comprises the K-Ras G12C mutation. In any of the above, the compound is Ras WT (For example, K-, H-, or N-Ras) WT ), or Ras amp (For example, K-, H-, or N-Ras) amp ) can also be inhibited in a similar manner.

[0085] Furthermore, a method for inhibiting intracellular Ras protein is also provided, which comprises contacting cells with an effective amount of the crystalline compound of the present invention. A method for inhibiting RAF-Ras binding is also provided, wherein the method comprises contacting cells with an effective amount of the crystalline compound of the present invention. The cells may be cancer cells. The cancer cells may be of any type of cancer described herein. The cells may be in vivo or in vitro.

[0086] 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 treatments (e.g., non-pharmacological treatments or therapeutic agents). The dosage of one or more of the additional treatments (e.g., non-pharmacological treatments or therapeutic agents) may be reduced from the standard dosage when administered alone. For example, the dosage may be determined empirically from the combination and permutation of drugs or estimated by isoborographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[0087] The crystalline form of the compound of the present invention may be administered before, after, or concurrently with one or more of such additional treatments. When combined, the dosage of the crystalline compound of the present invention and the dosage of one or more additional treatments (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 treatments such as anticancer agents may be administered together or separately in a single pharmaceutical composition, and if administered separately, this may occur simultaneously or sequentially. Such sequential administrations may be close together or far apart in time.

[0088] In some embodiments, an additional treatment is the administration of an adverse-limiting agent (e.g., an agent intended to reduce the occurrence or severity of adverse effects of the treatment). For example, in some embodiments, the crystalline compounds of the present invention may also be used in combination with a therapeutic agent for treating nausea. Examples of agents that can be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0089] In some embodiments, one or more additional treatments include non-pharmacological treatments (e.g., surgery or radiotherapy). In some embodiments, one or more additional treatments 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 treatments include non-pharmacological treatments (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 treatments include two therapeutic agents. In yet another embodiment, one or more additional treatments include three therapeutic agents. In some embodiments, one or more additional treatments include four or more therapeutic agents.

[0090] In this section on combination therapies, all references for the listed medications are incorporated by reference, whether explicitly stated or not. 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.

[0091] In some embodiments, the compounds of the present invention can be used as adjuvant therapy after surgery. In some embodiments, the compounds of the present invention can be used as preoperative adjuvant therapy before surgery.

[0092] 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 may be administered through 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. This term is intended to include, but not limited to, 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. In non-limiting examples, 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 gels or radioactive microspheres.

[0093] In some embodiments, the compounds of the present invention can make abnormal cells more sensitive to radiation therapy for the purpose of killing such cells or inhibiting their proliferation. Accordingly, the present invention further relates to a method for sensitizing abnormal cells in a mammal to radiation therapy, comprising administering a certain amount of the crystalline compound of the present invention to a mammal, the amount of which is effective for sensitizing abnormal cells to radiation therapy. The amount of 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 radiation therapy or as neoadjuvant therapy before radiation therapy.

[0094] 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 can 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. Prior to T cell proliferation and recombination, the T cell source is obtained from the subject. T cells can be obtained from several 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. Regardless of whether T cells have undergone genetic recombination to express a desired protein (e.g., CAR), T cells are generally, 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 7,1 It can be activated and propagated using the methods described in Nos. 44,575, 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.

[0095] 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 combined with a second, third, fourth, or more therapeutic agents. The crystalline compound of the present invention may be combined with one or more therapeutic agents in conjunction with one or more non-pharmacological therapies.

[0096] For example, the therapeutic agent may be a steroid. Steroids are known in the art. Therefore, in some embodiments, one or more additional therapeutic agents include steroids. Preferred steroids include 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, crocortol, cloprednol, corticosterone, cortisone, cortibazole, deflazacort, desonide, desoxymethasone, dexamethasone, diflorasone, diflucortol, difprednate, enoxolone, fluazacort, fluchloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, flucortin butyl, flucortolone, fluorometholone, fluperolone acetate, flupredniden acetate, fluprednisolone, flulandrenolide, Examples include, but are 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 their salts or derivatives.

[0097] Further examples of therapeutic agents that may be used in combination therapy with the crystalline compounds of the present invention include the following patents: U.S. Patents 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. Examples of compounds described in 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 include those described in applications WO01 / 37820, WO01 / 32651, WO02 / 68406, WO02 / 66470, WO02 / 55501, WO04 / 05279, WO04 / 07458, WO04 / 09784, WO02 / 59110, WO99 / 45009, WO00 / 59509, WO99 / 61422, WO00 / 12089, and WO00 / 02871.

[0098] The therapeutic agent may be a biological agent used to treat cancer or related conditions (e.g., cytokines (e.g., interferons or interleukins, e.g., IL-2)). Biological agents are known in the art. In some embodiments, the biological agent is an immunoglobulin biological agent, such as a monoclonal antibody (e.g., humanized antibody, fully human antibody, Fc fusion protein, or functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important to cancer. Antibody-drug conjugates are also included.

[0099] 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 of CTLA-4 (e.g., an anti-CTLA-4 antibody or fusion protein) (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist of PD-1 (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist of PD-L1 (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., a 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), a PD-L1 antibody such as avelumab, durvalumab, atezolizumab, pizilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or These are, but are not limited to, checkpoint inhibitors disclosed in Preusser, M. et al. (2015) Nat. Rev. Neurol., including ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514 / MEDI0680, BMS936559, MEDl4736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirirumab, IPH2101, 1-7F9, and KW-6002.

[0100] 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.

[0101] 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 in 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.

[0102] 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 docetaxel. In some embodiments, one or more additional treatments include two or more anticancer agents. Two or more anticancer agents can be used in a mixture and administered in combination or separately. Preferred 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).

[0103] Other non-exclusive examples of anticancer drugs include alkylating agents such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), thiotepa and cyclophosphamide, alkyl sulfonates such as busulfan, improsulfan and piposulfan, aziridines such as benzodopa, carboquan, metsuredopa and uredopa, altretamine and triethylenemelamine. Ethylenemimine and methylamelamamine, including triethylenetoformamide, triethylenetophosphoramide, and trimethylolmelamine, acetogenins (especially bratacin and bratacin), camptothecin (including the synthetic analog topotecan), bryostatin, callistatin, CC-1065 (including its synthetic analogs adzeresin, karzeresin, and bizeresin), cryptophycin (especially cryptophycin 1 and cryptophycin 8), dorastatin, duocalmycin (including synthetic analogs KW-2189 and CB1-TM1), erytherobin, pa Antibiotics such as cincratistatin, sarcodicin A, spongstatin, chlorambucil, chlornafadin, cyclophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine hydrochloride, melphalan, nobembicin, fenestrine, prednimustine, trophosphamide, uracil mustard and other nitrogen mustards, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine, nitrosoureas, engine antibiotics (e.g., Kalichemycin gamma II and Kalichemycin). Kaliceamines such as Amisin Omega II (see, for example, Agnew, Chem. Intl. 33:183-186 (1994)), dynemicins such as Dynemicin A, bisphosphonates such as clodronate, esperamicin, neocardinostatin chromophores and related pigment proteins, enediin antibiotic chromophores, acrasinomycin, actinomycin, autoramycin, azaserin, bleomycin, kakutinomycin, Kaliceamines, 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 such as mitomycin C, mycophenolate, nogaramycin, olibomycin, peplomycin, potophyllomycin, pew Antimetabolites such as romycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolubicin, methotrexate and 5-fluorouracil (5-FU), folate analogs such as denopterin, pteropterin and trimethrexate, purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine and thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, and doxif Pyrimidine analogs such as lurizine, enocitabine, and floxyuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; anti-adrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid solutions such as floric acid; acegraton, aldofamide glycoside, aminolevulinic acid, enyluracil, amsacrin, bestrabusil, bisantren, edatrexate, defofamine, deme Corsine, diaziquan, elfomithine, eriptinium acetate, epotilone B and other epotilones, etoglucide, gallium nitrate, hydroxyurea, lentinan, ronidynin, mytansinoids such as mytansin and anthamitosin, mitoglucone, mitoxantrone, mopidamol, nitracrine, pentostatin, fenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK® polysaccharide complex (JHS Natural Products, Eugene, OR), razoxane, rhizoxin, schizophyllan, spirogermanium, tenuazonic acid, triadicone, 2,2',2''-trichlorotriethylamine, T-2 toxin,Trichothecenes such as Veraculine A, Loridine A, and Anguidin, urethanes, vindesine, dacarbazine, mannomustine, mitobronitol, mitractol, pipobromane, gacytosine, arabinoside ("Ara-C"), cyclophosphamide, thiotepa, taxoids, e.g., Taxol® (paclitaxel), Abraxane® (a chromophore-free albumin-modified nanoparticle formulation of paclitaxel), and Taxotere® (docetaxel), chlorambucil, tamoxifen (Nolvadex®), raloxifen, aromatase inhibitory 4(5)-imidazole, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY 117018, Onapristone, Toremifene (Fareston®), Flutamide, Niltamide, Bicalutamide, Leuprolide, Goserelin, Chlorambucil, Gemzar® Gemcitabine, 6-Thiogunine, Mercaptopurine, Cisplatin, Oxaliplatin, and Platinum analogs such as Carboplatin, Vinblastine, Platinum, Etoposide (VP-16), Ifosfamide, Mitoxantrone, Vincristine, Navel Examples include bine® (vinorelbine), novantrone, teniposide, edatrexate, daunomycin, aminopterin, ibandronate, irinotecan (e.g., CPT-11), the topoisomerase inhibitor RFS2000, difluoromethylornithine (DMFO), retinoids such as retinoic acid, esperamicin, capecitabine (e.g., Xeloda®), and any pharmaceutically acceptable salts of the above.

[0104] Non-exclusive examples of additional anticancer drugs include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, Avisin, avagovomab, acridine carboxamide, adecatumumab, and 17-N-allyluane. Mino-17-demethoxygeldanamycin, alfarazine, arbocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracendione, anti-CD22 immunotoxin, antitumor agents (e.g., cell cycle nonspecific antitumor agents, and other antitumor agents described herein), antitumor herbs, apadiquon, atiprimod, azathioprine, berotecan, bendamustine, BIBW 2992, Bilicodal, Brostarisin, Briostatin, Butionine sulfoximine, CBV (chemotherapy), Calyculine, Dichloroacetate, Discordamorid, Elsamitolu, Enocitabine, Eribulin, Exatecan, Exislind, Ferginol, Forodesine, Phosfestrol, ICE chemotherapy regimen, IT-101, Imexone, Imiquimod, Indocarbazole, Ilofluben, Lanikidal, Lalotaxel, Lenalidomide, Lucanton, Lulutotecan, Maphosfamide, Mitozolomide, Examples include napoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, reximod, rubitecan, SN-38, salinosporamide A, sapacitabine, stanford V, swainsonin, talaporfin, talikidal, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, bajimezan, vinflunin, ZD6126, and zoskidal.

[0105] Further non-exclusive examples of anticancer drugs include vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mitramycin), mitomycin, and enzymes (e.g., those that systemically metabolize L-asparagine and combine it with its own asparagine). L-asparaginase, which eliminates cells that lack the ability to function, antiplatelet agents, antiproliferative / antimitotic alkylating agents such as nitrogen mustard (e.g., mechloretamine, cyclophosphamide and its analogs, melphalan, and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., CDK4 / 6 inhibitors such as abemaciclib, ribociclib, or palbociclib, sericiclib, UCN-01, P1446A-0) Antiproliferative / antimitotic metabolites and related inhibitors (e.g., 5, PD-0332991, Dynacyclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and its analogues, as well as streptozocin), trazeneth-dacarbadinine (DTIC), folic acid analogues, pyrimidine analogues (e.g., fluorouracil, floxuridine, and cytarabine), purine analogues, etc. (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum-coordinated complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroamic acid, vorinostat, LBH)589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., vistocertib, temsirolimus, everolimus, ridafololimus, and sirolimus), KSP(Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis®), PI3K inhibitors such as PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), copanlisib, alpelisib, and idelalisib, multikinase inhibitors (e.g., TG02 and sorafenib), hormone agonists such as hormones (e.g., estrogen) and 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-CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17 Examples include natural products such as AAG and KOS953, 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.

[0106] In some embodiments, the anticancer agent is selected from mechloretamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any of the aforementioned analogues or derivative variants.

[0107] 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 such as trastuzumab (Herceptin®) and pertuzumab (Perjeta®), small molecule tyrosine kinase inhibitors such as 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.

[0108] 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.

[0109] 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, SH3809, PF-07284892, or BBP-398) or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, These include SOS1 inhibitors (e.g., BI-1701963, BI-3406, SDR5, BAY-293, MRTX-0902, or RMC-5845), or pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers thereof), 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.

[0110] 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, WO20211 The compounds disclosed in 05960, WO2021074227, WO2020180768, WO2020180770, WO2020173935, WO2020146470, WO2019201848, WO2019122129, WO2018172250, and WO2018115380, or selected from their pharmaceutically acceptable salts, solvates, isomers, 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.

[0111] 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 an additional 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 ("Ras(OFF)"). As used herein, the term "Ras(OFF) inhibitor" refers to an inhibitor that targets the GDP-bound inactive state of Ras, i.e., an inhibitor that selectively binds to or inhibits it (e.g., selectively compared to the GTP-bound active state of Ras). Inhibition of the GDP-bound inactive state of Ras includes, for example, sequestering the inactive state by inhibiting the exchange of GDP to GTP, thereby preventing RAS from adopting the active conformation. In certain embodiments, the Ras(OFF) inhibitor may also bind to or inhibit the GTP-binding active state of Ras (for example, with a lower affinity or inhibition constant than the GDP-binding inactive state of Ras). In some embodiments, the Ras(OFF) inhibitor has a molecular weight of less than 700 Da. The term “KRas(OFF) inhibitor” refers to any Ras inhibitor that binds to KRas at the GDP-binding “OFF” position. References to the term KRas(OFF) inhibitor include, for example, AMG 510, MRTX849, JDQ443, and MRTX1133. In some embodiments, the KRas(OFF) inhibitor is selected from AMG 510 and MRTX849. In some embodiments, the KRas(OFF) inhibitor is AMG 510. In some embodiments, the KRAS(OFF) inhibitor is MRTX849. In some embodiments, the KRas(OFF) inhibitor is selected from BPI-421286, JNJ-74699157 (ARS-3248), LY3537982, MRTX1257, ARS853, ARS1620, and GDC-6036.

[0112] In some embodiments, the Ras inhibitor is an inhibitor of K-Ras G12C 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, 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. 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-KRAS(OFF) inhibitor. In specific embodiments, the pan-KRAS(OFF) inhibitor is JAB-23400, JAB-23425, BI-2493, BI-2865, QTX-3034 (G12D preference), QTX3544 (G12V preference), ZG2001, BBO-a, BBO-B, or pan-KRas-IN-1. In some embodiments, the Ras inhibitor is JAB-23400. In some embodiments, the Ras inhibitor is RMC-6236. In some embodiments, the Ras inhibitor is LUNA18. In some embodiments, the Ras inhibitor is BI-2493. In some embodiments, the Ras inhibitors are the Ras(ON) inhibitors disclosed in WO2023025832, WO2023015559, WO2022235870, WO2022235864, WO2021091982, WO2021091967, WO2021091956, and WO2020132597, which are incorporated herein by reference in their entirety.Alternatively, it may be selected from a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. The following are incorporated herein by reference in their entirety: WO2023287896, WO2023287730, WO2023284881, WO2023284730, WO2023284537, WO2023283933, WO2023283213, WO2023280960, WO2023280280, WO2023278600, WO2023280136, WO2023280026, WO2023278600, WO2023274383, WO2023274324, WO2023034290 , WO2023020523, WO2023020521, WO2023020519, WO2023020518, WO2023018812, WO2023018810, WO2023018809, WO2023018699, WO2023015559 , WO2023014979, WO2023014006, WO2023010121, WO2023009716, WO2023009572, WO2023004102, WO2023003417, WO2023001141, WO2023001123 , WO2022271923, WO2022271823, WO2022271810, WO2022271658, WO2022269508, WO2022266167, 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、WO2020028706、WO2019241157、WO2019232419、WO2019217691、WO2019217307、WO2019215203、WO2019213526、WO2019213516、WO2019155399、WO2019150305、WO2019110751、WO2019099524、WO2019051291、WO2018218070、WO2018217651、WO2018218071、WO2018218069、WO2018206539、WO2018143315, WO2018140600, WO2018140599, WO2018140598, WO2018140514, WO2018140513, WO2018140512, WO2018119183, WO201 8112420, WO2018068017, WO2018064510, WO2017201161, WO2017172979, WO2017100546, WO2017087528, WO2017058807, WO20170588 Other examples of Ras inhibitors, such as those in 05, WO2017058728, WO2017058902, WO2017058792, WO2017058768, WO2017058915, WO2017015562, WO2016168540, WO2016164675, WO2016049568, WO2016049524, WO2015054572, WO2014152588, WO2014143659, and WO2013155223, are known in the art.

[0113] In some embodiments, the therapeutic agent that can be combined with the crystalline compound of the present invention is RAS MULTI It is an (ON) inhibitor. When used herein, "RAS MULTI The term “(ON) inhibitor” refers to RAS(ON) inhibitors of at least three RAS variants having a missense mutation at one of the following positions: 12, 13, 59, 61, or 146. In some embodiments, RAS MULTI (ON) inhibitors refer to RAS(ON) inhibitors of at least three RAS variants having a missense mutation at one of the following positions: 12, 13, and 61. MULTI (ON) inhibitors are synthetic ligands (Ras MULTI The mechanism of action involves the formation of a high-affinity three-component complex between an (ON) inhibitor and two intracellular proteins that do not interact under normal physiological conditions (the target protein Ras and the cytoplasmic chaperone protein cyclophyllin A, which is widely expressed in cells), and the triple complex Ras MULTI (ON) inhibitor. Ras tricomplex. MULTINon-limiting examples of (ON) inhibitors include those disclosed in WO2021 / 091956 and WO2022 / 060836, or their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers.

[0114] In some embodiments, the therapeutic agent that may be combined with the crystalline compound of the present invention is an inhibitor of the MAP kinase (MAPK) pathway (or "MAPK inhibitor"). 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, rifametinib (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. PI3K / AKT inhibitors may include, but are not limited to, one or more PI3K / AKT inhibitors listed in Cancers (Basel) 2015 Sep;7(3):1758-1784. For example, a 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, and 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 pharmaceutically acceptable salts thereof. 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 native ligand. Non-exclusive 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] 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 the following patent publications: EP0520722, EP0566226, WO96 / 33980, U.S. Patent No. 5,747,498, WO96 / 30347, EP0787772, WO97 / 30034, WO97 / 30044, WO97 / 38994, WO97 / 49 688, EP837063, WO98 / 02434, WO97 / 38983, WO95 / 19774, WO95 / 19970, WO97 / 13771, WO9 8 / 02437, WO98 / 02438, WO97 / 32881, DE19629652, WO98 / 33798, WO97 / 32880, WO97 / 3288 Examples include any of the EGFR inhibitors listed in 0, EP682027, WO97 / 02266, WO97 / 27199, WO98 / 07726, WO97 / 34895, WO96 / 31510, WO98 / 14449, WO98 / 14450, WO98 / 14451, WO95 / 09847, WO97 / 19065, WO98 / 17662, U.S. Patent No. 5,789,427, U.S. Patent No. 5,650,415, U.S. Patent No. 5,656,643, WO99 / 35146, WO99 / 35132, WO99 / 07701, and WO92 / 20642, and all pharmaceutically acceptable salts of such EGFR inhibitors.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, for example, wortmannin, a 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]propionnitrile (BEZ 235 or NVP-BEZ Also known as 235, listed 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 (listed 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 (available from Axon Medchem), TGX-221(7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a] Pyrimidine-4-one (available from Axon Medchem), XL-765, and XL-147 are included, but are not limited to these. Other PI3K inhibitors include demethoxypyridine, perifosine, 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 and include 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)). This includes, but is not limited to, suppl):3493S-3498S), perifosine (e.g., 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 identifier: NSC 154020, Yang et al., Cancer Res. 2004, 64:4394-9).

[0122] mTOR inhibitors are known in the art and include ATP competitive mTORC1 / mTORC2 inhibitors, such as PI-103, PP242, PP30, Torin 1, FKBP12 enhancer, 4H-1-benzopyran-4-one derivatives, and rapamycin (also known as sirolimus) and its derivatives (Temsirolimus (Torisel®), Everolimus (Afinitor®, WO94 / 09010), Ridaforolimus (also known as dehorolimus or AP23573), for example, as disclosed in WO98 / 02441 and WO01 / 14387. For example, AP23464 and AP23841, 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxymethyl(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779), 40-epi(tetrazolito)-rapamycin (also known as ABT578), 32-deoxorapamycin, 16-pentinyloxy-32(S)-dihydrorapamycin, WO05 / 005434 The disclosed derivatives are 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, WO94 / 02485, WO95 / 14023, WO94 / 02136, and WO95 / 16 This includes, but is not limited to, derivatives disclosed in 691, WO96 / 41807, WO96 / 41807, and WO2018204416, as well as phosphorus-containing rapamycin derivatives (e.g., WO05 / 016252). In some embodiments, the mTOR inhibitor is a disteric inhibitor such as RMC-5552 (see, for example, WO2018204416, WO2019212990, and WO2019212991).

[0123] 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.

[0124] 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 to targeted therapies, including BCL-2 inhibitors such as ABT-263, as well as to conventional chemotherapy.

[0125] In some embodiments, additional therapeutic agents are SHP2 inhibitors. SHP2 is known in the art. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that 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, autoinhibitory 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 through receptor tyrosine kinases (RTKs) leads to exposure of the catalytic site, resulting in enzymatic activation of SHP2.

[0126] 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 either SHP2 autoactivation or enhanced growth factor-driven activation. Therefore, SHP2 represents 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 with SHP2 inhibitors along with RAS pathway inhibitors may be a common strategy for preventing tumor resistance in a wide range of malignancies.

[0127] Non-limiting examples of such SHP2 inhibitors known in the art include 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., each incorporated herein by reference. al., Oncotarget, 2017, 8, 113734, and Patent Publications: WO2023282702, WO2023280283, WO2023280237, WO2023018155, WO2023011513, WO2022271966, WO2022271964, WO2022271911, WO2022259157, WO2022242767, WO2022241975, WO2022237676, WO20222373 67, WO2022237178, WO2022235822, WO20222084008, WO2022135568, WO2021176072, WO2021171261, WO2021149817, WO20211 48010, WO2021147879, WO2021143823, WO2021143701, WO2021143680, WO2021121397, WO2021119525, WO2021115286, WO2021 110796, WO2021088945, WO2021073439, WO2021061706, WO2021061515, WO2021043077, WO2021033153, WO2021028362, WO20 21033153, WO2021028362, WO2021018287, WO2020259679, WO2020249079, WO2020210384, WO2020201991, WO2020181283, WO2 020177653, WO2020165734, WO2020165733, WO2020165732, WO2020156243, WO2020156242, WO2020108590, WO2020104635, W O2020094104, 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、Examples include CN108113848, US11179397, US11044675, US11034705, US11033547, US11001561, US10988466, US10954243, US10934302, or US10858359, or their pharmaceutically acceptable salts, solvates, isomers (e.g., stereoisomers), prodrugs, or tautomers.

[0128] 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 an 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.

[0129] In some embodiments, the SHP2 inhibitor has the following structure:

[0130] [ka]

[0131] The SHP2 inhibitor is TNO155, or 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 has the structure:

[0132] [ka]

[0133] RMC-4630 having the structure: or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor has the structure:

[0134] [ka]

[0135] The SHP2 inhibitor is JAB-3068, or 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] or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor has the following structure:

[0138] [ka]

[0139] The inhibitor is RLY-1971, or 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 includes administration of 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 includes administration of 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 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., BiTEs), and anti-PD-1, anti-PD-L1, anti-CTLA4, anti-LAG1, 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. The IMiD class includes 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 / 121168A1, as well as elsewhere in this specification.

[0145] FGFR inhibitors, including FGFR2 and FGFR4 inhibitors, such as pemigatinib and erdafitinib, are known in the art. 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) such as GITR fusion proteins described in U.S. Patent No. 6,111,090, U.S. Patent No. 8,586,023, WO2010 / 003118, and WO2011 / 090754, or, for example, U.S. Patent No. 7,025,962, EP1947183, U.S. Patent No. 7,812,135, U.S. Patent No. 8,388,967, and U.S. Patent No. 8,59 This includes, but is not limited to, the anti-GITR antibodies 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 and conjugates thereof, which are synthetically prepared in vitro. Anti-angiogenic agents may be agonists, antagonists, allosteric modulators, toxins, or more generally, act to inhibit or stimulate their targets (e.g., by activating or inhibiting receptors or enzymes), thereby promoting cell death or inhibiting cell proliferation. In some embodiments, one or more additional therapeutic agents 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 / 528. As described in 89, 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 are those with little or no activity to inhibit MMP-1. More preferred are those that selectively inhibit MMP-2 or AMP-9 in relation 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 RS13-0830.

[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-Ang1 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 Campath, 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 disintegrin 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, 5,728,813, 5,969,110, 6,596,852, 6,232,447, 6,057,124, and their respective 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 (GileadSciences, USA), Alpha-Statin (BioActa, UK), M-PGA (Celgene, USA, US5712291), Ilostat (Arriva, USA, US5892112), Emaxanib (Pfizer, USA, US5792783), Batananib (Novartis, Switzerland), 2-Methoxyestradiol (EntreMed, USA), TLC ELL-12 (Elan, Ireland), Anecoltab Acetate (Alcon, USA), Alpha-D148 Mab (Amgen, USA), CEP-7055 (Cephalon, USA), Anti-Vn Mab (Crucell, Netherlands), DAC Anti-Angiogenic Agent (ConjuChem, Canada), Angiocidin (InKine Pharmaceutical, USA), KM-2550 (Kyowa Hakko (Japan), SU-0879 (Pfizer, USA), CGP-79787 (Novartis, Switzerland, EP 0970070), 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), XL647 (Exelixis, USA), MAb, second-generation alpha-5 beta-3 integrins (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), sirengitide (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 Childrens 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), xanthrizole (Yonsei University, South Korea), gene-based vaccine, VEGF-2 (Scripps Clinic and Research Foundation, USA), SPV5.2 (Supratek, Canada), SDX 103 (University of California at SanDiego, 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), angiogenesis vaccine (EntreMed, USA), urokinase plasminogen activator inhibitor (Dendreon, USA), ogluphanide (pINN) (Melmotte, USA), HIF-1-alpha 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), CP 547632 (Pfizer, USA), 786034 (GlaxoSmithKline, UK), KRN 633 (Kirin Brewery, Japan), Intraocular drug delivery system, 2-methoxyestradiol, Anginex (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), Combretastatin 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), AG 13925 (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), Tumustatin (Beth Israel Hospital, USA), Severable Soluble FLT 1 (Vascular Endothelial Growth Factor Receptor 1) (Merck & Co, USA), Tie- Examples include ligand 2 (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. In addition, antisense or siRNAs that inhibit the expression of proteins, including but not limited to ATG5 (which is involved in autophagy), may also be used. In some embodiments, one or more additional therapeutic agents include an autophagy inhibitor.

[0155] Another example of a therapeutic agent that can be used in combination with the crystalline compounds of the present invention is an antitumor agent known in the art. In some embodiments, one or more additional therapeutic agents include an antitumor agent. Non-limiting examples of antitumor 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 DA 3030 (Dong-A), daclizumab, denileukin difutox, 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 / Gafour combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, 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, LC9018 (Yakult), Leflunomide, Lenograstim, Lentinan sulfate, Letrozole, Leukocyte alpha interferon, Leuprorelin, Levamizole + Fluorouracil, Rialozol, Lovaplatin, Ronidamin, Lovastatin, Masopropyl, Melalsoprole, Metoclopramide, Mifepristone, Miltefosine, Millimostim, Mispaired double-stranded RNA, Mitoguazone, Mitractol, Mitoxantrone, Morglamostim, Nafarelin, Naloxone + Pentazocine, Naltgrastim, Nedaplatin, Niltamide, Noscapine, Novel Erythropoiesis-Promoting Protein, NSC631570 Octreotide, Oprelbequin, Osateron, Oxaliplatin, Paclitaxel, Pamidronic Acid, Pegaspargase, Peginterferon Alpha-2b, Pentosan, Sodium Polysulfate, Pentostatin, Picibanil, Pirarubicin, Rabbit Antithymocyte Polyclonal Antibody, Polyethylene Glycol Interferon Alpha-2a, Porfimer Sodium, Raloxifene, Larcitrexed, Rasbrien Bodymen Rasburiembodiment, rhenium etidronate Re186, RII retinamide, rituximab, romultide, samarium (153Sm) lexidonam, salglamostim, schizophyllan, sobuzoxane, sonelmin, strontium-89 chloride, suramin, tasonelmin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfacin, thyroid-stimulating hormone alpha, topotecan, toremifene, tositumomab-iodine-131, trastuzumab, treosulf Pharma, tretinoin, trilostane, trimethrexate, triptorelin, tumor necrosis factor alpha, natural type, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma solubilizing solution vaccine, barrubicin, verteporfin, vinorelbine, bilirulysine, dinostatin stimulamer or zoledronic acid, abalelix, AE941 (Aeterna), ambamustin, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), decitabine, dexaaminoglutethimide, diazicon, EL532 (Elan),EM800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, gallocitabine, gastrin-17 immunogene, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxyfen, LDI200 (Milkhaus), religistim, lintuzumab, CA125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine-131MAb (Techniclone), polymorphoemic mucin-yttrium-90MAb (Antisoma), marimast, menogalil, mitumomab, motexafine, gadolinium, MX6 (Galderma), nelarabine, noratexed, P30 protein, pegvisomant, pemetrexed, porphyromycin, prinomast, RL0903 (Shire), rubitecan, satoraplatin, sodium phenylacetate, sparphosic acid, SRL172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, saliblastin, thrombopoietin, tin ethylethiopurine, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Examples include the melanoma tumor lysis product vaccine (New York Medical College), the viral melanoma cell solubilization vaccine (Royal Newcastle Hospital), or Valspodar.

[0156] Additional examples of therapeutic agents that may 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 RG7446, MEDI-570, AMG557, MGA271, IMP321, BMS-663513, PF-05082566, CDX-1127, and anti-OX40 (Providence Health). Services), huMAbOX40L, Atacisep, CP-870893, Lucatumumab, Dacetuzumab, Muromonab-CD3, Ipilumumab, MEDI4736 (Imfinzi®), MSB0010718C, AMP224, adalimumab (Humira®), ado-trastuzumab emtansine (Kadcyla®), aflibercept (Eylea®), alemtuzumab (Campath®), basiliximab (Simulect®), belimumab (Benlysta®), basiliximab (Simulect®), belimumab (Benlysta®), brentuximab Buvedotin (Adcetris®), Canakinumab (Ilaris®), Certolizumab pegol (Cimzia®), Daclizumab (Zenapax®), Daratumumab (Darzalex®), Denosumab (Prolia®), Eculizumab (Soliris®), Efalizumab (Raptiva®), Gemtuzumab Ozogamicin (Mylotarg®) (Registered Trademarks)), Golimumab (Simponi®), Ibritumomab tiuxetan (Zevalin®), Infliximab (Remicade®), Motavizumab (Numax®), Natalizumab (Tysabri®), Obinutuzumab (Gazyva®), Ofatumumab (Arzerra®), Omalizumab (Xolair®), Palivizumab (Synagi®) Examples include s(registered trademark), pertuzumab (Perjeta(registered trademark)), pertuzumab (Perjeta(registered trademark)), ranibizumab (Lucentis(registered trademark)), laxibakumab (Abthrax(registered trademark)), tocilizumab (Actemra(registered trademark)), tositumomab, tositumomab-i-131, tositumomab and tositumomab-i-131(Bexxar(registered trademark)), ustekinumab (Stelara(registered trademark)), 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 therapeutic agents as described herein. When used in combination therapy, the compounds described herein may be administered simultaneously with or separately from a second agent. This combination administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration in different dosage forms, and separate administrations. That is, the crystalline compounds described herein and any of the agents described herein can be formulated together in the same dosage form and administered simultaneously. Alternatively, the crystalline compounds of the present invention and any of the therapeutic agents described herein can be administered simultaneously, with both agents present in separate formulations. Another alternative is that the crystalline compounds of this disclosure may be administered, followed by the administration of any of the therapeutic agents described herein, or vice versa. In some embodiments of separate administration protocols, the crystalline compounds of the present invention and any of the therapeutic agents described herein are administered at intervals of minutes, hours, or days.

[0158] In some embodiments of the methods described herein, the first treatment (e.g., the compound of the present invention) and one or more additional treatments are administered simultaneously, sequentially, or in any order. The first treatment agent may be administered immediately before or after one or more additional treatments, 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, 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 features a kit comprising (a) a pharmaceutical composition comprising an agent described herein (e.g., a crystalline compound of the present invention), (b) one or more additional treatments (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 using a combination of pharmaceutically active compounds that can be administered separately, further relates to combining separate pharmaceutical compositions in kit form. The kit may comprise two separate pharmaceutical compositions, namely the crystalline compound of the present invention and one or more additional therapeutic agents. 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 the use of 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 the prescribing healthcare professional desires to increase or decrease the individual components of the combination.

[0161] The following embodiments further illustrate the present invention, 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 construed as limiting 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 impose 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 available, which themselves may be suggested to those skilled in the art, without departing from the spirit of this disclosure or the appended claims.

[0163] Example 1 This example demonstrates an exemplary method for preparing crystalline form 1 of compound A according to embodiments of the present invention. Crystalline form 1 is prepared via precipitation using antisolvent addition, spontaneous precipitation in a solvent or mixture of solvents, evaporation of a solvent or mixture of solvents, and spontaneous crystallization in a solvent or mixture of solvents. Any of the methods described herein may also produce mixtures of crystalline forms 1 and 2 of compound A.

[0164] In one method, compound A was dissolved in isopropyl ether in a vial. To this mixture, a certain amount of ethanol was added so that the mixture yielded a 1:17 ratio of ethanol to isopropyl ether. The vial was loosely capped and stored under ambient conditions, yielding a precipitate of translucent crystals of Form 1. The crystals were isolated and dried. These crystals were used for X-ray crystallography to produce the crystalline structure of Form 1 as a mixed isopropyl ether, ethanol, and aqueous solvate.

[0165] Alternatively, compound A was dissolved in sufficient diethyl ether to produce a saturated slurry in a glass vial. The slurry was heated to 40°C and magnetically stirred to produce a solid. The crystals were isolated and dried. These crystals were used for X-ray crystallography to produce the crystal structure of Form 1 as a mixed diethyl ether and aqueous solvate.

[0166] Alternatively, 20.6 mg of compound A was dissolved in 0.5 mL of 2-butanol in a drum vial. The open vial was placed inside a 20 mL vial containing 2 mL of isopropyl ether. The outer vial was capped to allow vapor diffusion. After combined storage at room temperature and 8°C for approximately 3 weeks, the sample remained as a clear solution. Isopropyl ether (5 mL) was added, and the solution was magnetically stirred at approximately 8°C (refrigerator). After 1-2 days, the precipitate was observed, and the sample was further stirred at approximately 15°C (freezer) for 3 days to maximize the yield. The white solid was separated by centrifugation, and the remaining solvent was removed via pipette. The solid was dried in a vacuum desiccator for 0.5 hours and analyzed by XRPD analysis.

[0167] Alternatively, 23.1 mg of compound A was dissolved in 0.5 mL of 1-pentanol in a drum vial. The open vial was placed inside a 20 mL vial containing 2 mL of isopropyl ether. The outer vial was capped to allow vapor diffusion. After combined storage at room temperature and 8°C for approximately 3 weeks, the sample remained as a clear solution. Isopropyl ether (5 mL) was added, and the solution was magnetically stirred at approximately 8°C (refrigerator). After 1-2 days, the precipitate was observed, and the sample was further stirred at approximately 15°C (freezer) for 3 days to maximize the yield. The white solid was separated by centrifugation, and the remaining solvent was removed via pipette. The solid was dried in a vacuum desiccator for 0.5 hours and analyzed by XRPD analysis.

[0168] Alternatively, 21.0 mg of compound A was dissolved in 0.5 mL of ethyl acetate in a drum vial. The open vial was placed inside a 20 mL vial containing 2 mL of isopropyl ether. The outer vial was capped to allow vapor diffusion. After combined storage at room temperature and at 8°C for approximately 13 days, a viscous oily substance formed in the clear solution. The oil crystallized during additional storage at room temperature (approximately 11 days), producing a white solid. The solid was separated by centrifugation, and the remaining solvent was removed via pipette. The solid was dried in a vacuum desiccator for 0.5 hours and analyzed by XRPD analysis.

[0169] Alternatively, 21.0 mg of compound A was dissolved in 1.4 mg of acetic acid and 2 mL of diethyl ether, yielding a clear solution. The mixture was magnetically stirred overnight at room temperature. A white solid was observed the following day. The sample was centrifuged and the mother liquor was decanted. The isolated solid was dried in a fume hood.

[0170] Alternatively, 20.0 mg of compound A was dissolved in 2.5 mg of benzoic acid and 2 mL of diethyl ether, yielding a clear solution. The mixture was magnetically stirred at room temperature for 3 days. A white solid was observed. The sample was centrifuged and the mother liquor was decanted. The isolated solid was dried in a fume hood.

[0171] Alternatively, 20.0 mg of compound A was dissolved in 1.6 mg of glycolic acid and 2 mL of diethyl ether, yielding a clear solution. The mixture was magnetically stirred at room temperature for 4 days. A white solid was observed. The sample was centrifuged and the mother liquor was decanted. The isolated solid was dried in a fume hood.

[0172] Alternatively, 20.0 mg of compound A was dissolved in 4.1 mg of D,L-lactic acid and 2 mL of diethyl ether, yielding a clear solution. The mixture was magnetically stirred at room temperature for 3 days. A white solid was observed. The sample was centrifuged and the mother liquor was decanted. The isolated solid was dried in a fume hood.

[0173] Alternatively, approximately 20 mg of amorphous compound A was equilibrated in 1:1 v:v MeOH / water at 25°C for one week using a stirring rod on a magnetic stirring plate 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 1.

[0174] Alternatively, approximately 20 mg of amorphous compound A was dissolved in approximately 0.1 mL of 1:1 v / v acetone / water at ambient temperature (20-25°C). To this mixture, 0.22 mL of water was slowly added until a large amount of solid precipitated. The solid was collected by centrifugation filtration through a 0.45 μm nylon membrane filter at 14,000 rpm to obtain crystalline form 1.

[0175] Alternatively, crystal morphology 1 underwent a variable humidity XRPD experiment. In this experiment, two relative humidity (RH) cycles were applied at 25°C. XRPD analysis was performed for each specific relative humidity. Cycle 1: 40%RH (initial) - 40%RH (3 hours) - 60%RH (3 hours) - 80%RH (3 hours) - 95%RH (3 hours) - 80%RH (3 hours) - 60%RH (3 hours) - 40%RH (3 hours) - 20%RH (3 hours) - 0%RH (3 hours), Cycle 2: 20%RH (3 hours) - 40%RH (3 hours). When the relative humidity was higher than 80%RH, morphology 1 was converted or partially converted to morphology 2, and then morphology 2 was converted back to morphology 1 when the relative humidity was lower than 80%.

[0176] Alternatively, approximately 300 mg of amorphous compound A was weighed into an 8 mL glass vial. 2.4 mL of 1:1 v:v MeOH / water was added to the vial while stirring at 300–400 rpm at 25°C for 4 days. After stirring at 25°C for 4 days, the resulting suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid was dried under ambient conditions for approximately 12 hours. Approximately 221.13 mg of crystalline form 1 was obtained as a white powder in a yield of 71.16%.

[0177] Example 2 This example demonstrates an exemplary method for preparing a mixture of crystalline forms 1 and 2 of compound A according to an embodiment of the present invention. Any of the methods described in Example 1 may also produce a mixture of forms 1 and 2.

[0178] In one method, 200 mg of compound A was dissolved in hexane. To this mixture, a certain amount of ethyl acetate was added so that the mixture yielded a 1:2 ratio of ethyl acetate to hexane. The resulting mixture formed a slurry, which was stored at room temperature for 3 days and then placed in a vacuum oven at 40°C for 1.5 hours. The resulting solid was characterized by XRPD and identified as a mixture of forms 1 and 2 of compound A.

[0179] Example 3 This example demonstrates the X-ray powder diffraction (XRPD) characterization of a single crystalline form 1 of compound A and a mixture of crystalline forms 1 and 2 of compound A according to embodiments of the present invention. The X-ray powder diffractogram of form 1 as a mixed ethanol and isopropyl ether solvate is shown in Figure 1. In the essentially pure substance of form 1 as a mixed ethanol and isopropyl ether solvate, peaks can be observed at a refraction angle 2θ, as shown in Table 1.

[0180] [Table 1-1]

[0181] [Table 1-2]

[0182] The X-ray powder diffractograms of the mixtures of forms 1 and 2 are shown in Figure 2. In the crystalline samples of forms 1 and 2, peaks can be observed at a refraction angle of 2θ, as shown in Table 2.

[0183] [Table 2-1]

[0184] [Table 2-2]

[0185] As described in Example 1, a method for producing form 1 of compound A can produce a mixture of form 1 and 2 of compound A, having varying relative peak intensities observed by XRPD analysis, suggesting various ratios of the two forms. The formation of form 2 is indicated by the presence of a strong peak at 4.8°2θ (Figure 2), which is not present in a pure sample of form 1 (Figures 1 and 3). To study the formation of form 2, a saturated slurry of compound A was prepared in diethyl ether, which produced pure form 1 at time = 0 hours, and this pure sample was monitored over time using XRPD analysis. A shoulder peak at 4.8°2θ was detected at approximately 2 hours and increased in intensity compared to the original form 1 peak after 4 and 17 days (Figure 4). Thus, a pure sample of form 1 can produce a mixture of form 1 and 2 over time, as described in Example 1.

[0186] Example 4 This example demonstrates the single-crystal X-ray crystallographic characterization of crystalline form 1 of free base compound A according to an embodiment of the present invention. The X-ray crystal structure of crystalline form 1 of compound A as a mixed isopropyl ether, ethanol, and aqueous solvate (chiral unit) is shown in Figure 5.

[0187] It has approximate dimensions of 0.16 × 0.14 × 0.01 mm, as shown in formula 4(C 55 H 78 FN9O8)·3(C6H 14A colorless crystal of form 1 having O)·2(C2H6O)·2(H2O) was mounted on a Mitegen micromesh mount in a random orientation. Preliminary testing and data acquisition were performed using Cu Kα radiation (λ=1.54178 Å) on a Bruker AXS D8 Quest CMOS diffractometer equipped with a 4-axis copper stage, an I-μ-S microsource X-ray tube transverse gradient multilayer optics system, a PhotonIII-C14 single-photon count detector, and an Oxford Cryosystems cryogenic device. The first unit cell was determined and data was acquired using Apex3 v2019.11-0 at a temperature of 150K. Frames were integrated using SAINT V8.40B. A total of 61,485 reflections were collected, of which 23,916 were unique. Cell constants for data acquisition were obtained from least-squares refinement using 6,855 reflections between 2.2752 and 58.3702°. The orthogonal cell parameters and calculated quantities are a = 40.5965 (16) Å, b = 16.0423 (5) Å, c = 19.4198 (9) Å, and V = 12,647.4 (9) Å. 3 Therefore, for the formula weights Z=2 and 4483.72, the calculated density is 1.177 g / cm³. 3 The linear absorption coefficient was 0.665 / mm for Cu Kα radiation. Scaling and multiscan absorption correction were applied using SADABS 2016-2. The transmission coefficient ranged from 0.6125 to 0.7543. The intensity of equivalent reflection was not averaged during data processing.

[0188] The space group was determined using the XPREP program integrated into SHELXTL. The intensity statistics showed the space group P21212(#18). The structure was decomposed by isomorphic substitution from its diethyl ether solvate, and all reflections were used with SHELXL-2018 and the graphical user interface ShelXle to determine the F 2 The matrix was refined by the total least squares method. Additional atoms were identified in the subsequent difference Fourier synthesis. The minimized function is Σw(|F o | 2 -|F c | 2) 2 And the weight w is w = 1 / [σ 2 (F o 2 )+(0.0866P) 2 ] is defined as, in the formula, P=(F o 2 +2F c 2 The structure was refined using the total matrix least squares method, which is ) / 3. Scattering factors were determined from the International Tables for Crystallography (Vol C Tables 4.2.6.8 and 6.1.1.4). A total of 25,975 independent reflections were used for refinement. F 2 >2σ(F 2 10,446 reflections with the following characteristics were used to calculate R1.

[0189] Two crystallographically independent molecules exist in the structural lattice. A general atomic naming scheme was used, and the suffixes A and B were added to distinguish the molecules. The hydrogen atoms bonded to carbon were geometrically positioned and constrained to rest on their parent atoms. CH bond distances were constrained to 0.95 Å for aromatic and alkene CH moieties, and to 1.00, 0.99, and 0.98 Å for aliphatic CH, CH2, and CH3 moieties, respectively. Methyl hydrogen atoms were initially rotated to best suit the experimental electron density. Some hydrogen atoms in disordered methyl groups were set to alternate positions in the final elaboration cycle. Amine and amide hydrogen atom positions were refined, restricting the NH distance to 0.88(2) Å. Alcohol OH bond distances were initially constrained to 0.84° but rotated to best suit the experimental electron density. Water hydrogen atom positions were initially refined, restricting the OH and H...H distances to 0.84(2) and 1.36(2) Å, respectively. Where necessary, water hydrogen atom positions were further restricted based on hydrogen bonding considerations (see the following sections for details). In the final refinement cycle, the positions of the water and alcohol H atoms were set to align with the positions of their carrier O atoms. isoThe (H) values ​​are 1.5 times for OH and CH3, and 1.2 times for CH, CH2, and NH units. eq It was set to (C).

[0190] For molecule A, the methoxymethyl group was refined as disordered. The major and secondary OC bonds were restricted to have similar lengths. The ADP of the O and C atoms ij The components were restricted to be similar. Under these conditions, the occupancy ratio was refined to 0.649(15) to 0.351(15).

[0191] For molecule B, disruption of the N,N-dimethylpropane-2-amine substituent is observed. The fragment was refined as disordered across three alternative orientations (suffixes B, C, and D). The three disordered parts were restricted to have a geometric shape similar to the non-disordered equivalent fragment of molecule A. The U of ADP for disordered atoms closer to each other than 2.0 Å ij The components were restricted to be similar. Under these conditions, the occupancy ratios were refined to 0.471(4), 0.241(4), and 0.288(4) for N,N-dimethylpropan-2-amine moieties B, C, and D, respectively.

[0192] A single, fully occupied water molecule (related to O1) is located on the double rotation axis, while nearby ethanol molecules are in a 1:1 disorder around the same double rotation axis. The water molecule acts as a hydrogen bond acceptor for two symmetric equivalent NH...O hydrogen bonds (including the amide N4B), as well as a hydrogen bond donor for two disordered solvate ethanol molecular parts (oxygen O3) and either O3B or its symmetric equivalent by double rotation, thus inducing a 1:1 obstruction for the water H atom. The O...H hydrogen bond distance was initially restricted to 2.20(2) Å (from H1O1 to O3B, and from H1O2 to O2), and the distance between H1O1 and H4NB (of the amide N4B) was restricted to at least 2.30(2) Å. In the final refinement cycle, the positions of the water and alcohol H atoms were set to rest on the positions of their carrier O atoms. The ethanol OC and CC bond distances were restricted to the expected target values ​​(1.430(1) and 1.53(2) Å, respectively) and were also restricted to be similar to those of another ethanol solvate molecule. ij The ingredients were restricted to be similar.

[0193] The diisopropyl ether molecule (related to O2) exhibits large vibrations, which may indicate a malfunction, but it is not well-defined to develop a meaningful malfunction model. The extended channels, located near both a single diisopropyl ether molecule and a disordered N,N-dimethylpropane-2-amine fragment and bifurcated by a double axis, were refined so that they were occupied by disordered diisopropyl ether and ethanol molecules, respectively, with half the occupancy (imposed by the double axis). The ethanol molecules consist of O6B, the ethanol molecule, and a half-occupied water molecule hydrogen-bonded to the amine N atom N9B or N9C. The disordered diisopropyl ether molecule was restricted to be geometrically similar to the other fully occupied diisopropyl ether molecule. The ethanol OC and CC bond distances were restricted to expected target values ​​(1.430(1) and 1.53(2) Å, respectively), and also restricted to be similar to those of the other ethanol solvate molecule.

[0194] The final refinement cycle, including 1,688 variable parameters and 628 constraints, converged with the following unweighted and weighted coincidence coefficients (the maximum parameter shift was 0.003 times its standard uncertainty): R1 = Σ|F o |-|F c | / Σ|F o |=0.0727 wR 2 ={Σ[w(F o 2 -F c 2 )2] / Σ[w(F o 2 ) 2 ]} 0.5 =0.2142 The goodness-of-fit parameter was 0.949. The highest peak in the final difference Fourier map was 0.351 e / Å. 3 It had a height of -0.353 e / Å. The smallest negative peak was -0.353 e / Å. 3 It had a height of [value missing]. Crystal data and data acquisition parameters are listed in Table 4.

[0195] [Table 3-1]

[0196] [Table 3-2]

[0197] Example 5 This example demonstrates the single-crystal X-ray crystallographic characterization of crystalline form 1 of compound A according to an embodiment of the present invention. The X-ray crystal structure of form 1 as a mixed diethyl ether and aqueous solvate (chiral unit) is shown in Figure 6.

[0198] Formula C has approximate dimensions of 0.13 × 0.08 × 0.03 mm. 55 H 78 FN9O8·1.086(C4H 10Beige crystals of form 1 having O)·0.35(H2O) were mounted on a Mitegen micromesh mount in random orientation. Preliminary testing and data acquisition were performed using Cu Kα radiation (A=1.54178A) on a Bruker AXS 08 Quest CMOS diffractometer equipped with a 4-axis copper stage, an IpS microsource X-ray tube transverse gradient multilayer optics system, a PhotonIII-C14 single-photon count detector, and an Oxford Cryosystems cryogenic device. The first unit cell was determined and data was acquired using Apex3 v2019.11-0 at a temperature of 150K. Frames were integrated using SAINT V8.40B. A total of 81,435 reflections were collected, of which 25,975 were unique. Cell constants for data acquisition were obtained from least-squares refinement using 9,983 reflections ranging from 2.5549 to 75.91130. The orthogonal cell parameters and calculated quantities are a=40.813(8)A, b=16.079(4)A, c=19.093(4)A, and V=12,529(4)A. 3 The calculated density for the formula weights Z=8 and 1099.06 is 1.165 g / cm³. 3 The linear absorption coefficient is 0.659 / mm for Cu Kα radiation. Scaling and multiscan absorption correction were applied using SADABS 2016-2. The transmission coefficient ranged from 0.6883 to 0.7543. The intensity of equivalent reflection was not averaged during data processing.

[0199] The space group was determined using the XPREP program integrated into SHELXTL. The intensity statistics showed the space group P21212(#18). The structure was decomposed using a direct method with SHELXM (Sheldrick, 2008), and all reflections were used with F using SHELXL-2018 and the graphical user interface ShelXle. 2 The matrix was refined by the total least squares method. Additional atoms were identified in the subsequent difference Fourier synthesis. The minimized function is Σw(|F o | 2 -|F c | 2) 2 where the weight w is w = 1 / [σ 2 (F o 2 )+(0.0637P) 2 +0.782P], where P = (F o 2 +2F c 2 ) / 3. The structure was refined using full-matrix least squares. The scattering factors were obtained from the International Tables for Crystallography (Vol C Tables 4.2.6.8 and 6.1.1.4). A total of 25,975 independent reflections were used in the refinement. 18,986 reflections with F 2 >2σ(F 2 ) were used in the calculation of R1.

[0200] Two crystallographically independent molecules are present in the lattice of the structure. A general atomic naming scheme was used, and suffixes A and B were added to distinguish the molecules. H atoms bonded to carbon were geometrically positioned and constrained to ride on their parent atoms. The C-H bond distances were constrained to 0.95 Å for aromatic and alkene C-H moieties, 1.00, 0.99, and 0.98 Å for aliphatic C-H moieties, CH2 moieties, and CH3 moieties, respectively. The amine and amide H atom positions were refined, and the N-H distance was restricted to 0.88(2) Å. The water H atom positions were refined, and the O-H and H...H distances were restricted to 0.84(2) and 1.36(2) Å, respectively. Where necessary, the water H atom positions were further restricted based on considerations of hydrogen bonding (see the following section for details). U iso (H) values were set to 1.5 times for CH3 and 1.2 times for C-H, CH2, and N-H units of U eq (C / N), respectively.

[0201] For molecule B, interference with the N,N-dimethylpropane-2-amine substituent is observed. The fragment was refined as disordered across three alternative orientations (suffixes B, C, and D). The three disordered parts were restricted to have a geometric shape similar to the non-disordered equivalent fragment of molecule A. Partially occupied water molecules (related to O7) are related to interference and are incompatible with some of the disordered fragments as well as some of their symmetry equivalent counterparts by the crystallographic double axis. Intrinsic assignment of water molecules to only one part is not possible, and therefore, its occupation was refined independently. The position of the water H atom was restricted based on consideration of hydrogen bonding, and the distances from H7O1 to N9B and H7O2 to O3B (of the major N,N-dimethylpropane-2-amine fragments in 2-x, -1-y, +z) were restricted to 2.10(2) and 2.20(2)A, respectively. The U of ADP for disordered atoms closer to each other than 2.0A ij The components were restricted to be similar. Under these conditions, the occupancy rates were refined to 0.583(4), 0.137(4), and 0.280(4) for N,N-dimethylpropan-2-amine moieties B, C, and D, respectively, and to 0.200(10) for water molecules.

[0202] A single, fully occupied water molecule (related to O3) is located on the double rotation axis. It acts as a hydrogen bond acceptor for two symmetric equivalent NH...O hydrogen bonds (including the amide N4B), as well as a hydrogen bond donor for either the solvate ethanol molecular portion (oxygen O2) and its symmetric equivalent by O3B or double rotation, thus inducing a 1:1 obstruction for the water H atom. The O...H hydrogen bond distance is limited to 2.20(2)A (from H1O1 to O3B, and from H1O2 to O2), and the distance between H1O1 and H4NB (of the amide N4B) is limited to at least 2.30(2)A. The ethyl group of the hydrogen of the ether molecule bonded to the water molecule is refined as a 1:1 disorder (the oxygen atom is located on the double axis). The ether OC and CC bond distances, as well as the O...O 1,3 distance (i.e., OCC angle), were limited to expected target values ​​(1.43(2), 1.53(2), and 2.48(2)A, respectively).

[0203] A single ether molecule (related to O3) exhibits large vibrations and is an indication of a disturbance, but it is not well defined, making it impossible to develop a meaningful disturbance model. An extended channel bifurcated by a double axis, located near both a single diethyl ether molecule and a disordered N,N-dimethylpropane-2-amine fragment, was refined as being occupied by the disordered diethyl ether molecule. Three crystallographically distinct molecules were defined (related to O4, O5, and O6). The majority of the three fragments (the O5 fragment) overlap with their symmetry equivalents by double rotation. For both single and disordered diethyl ether molecules, the OC and CC bond distances, as well as the O...C 1,3 distance (i.e., OCC angle), were again restricted to expected target values ​​(1.43(2), 1.53(2), and 2.48(2)A, respectively). Based on these conditions, the occupancy rates were refined to 0.163(4)(04), 2×0.328(2)(05), and 0.181(3)(06).

[0204] The final cycle of refinement included 1721 variable parameters and 781 restraints and converged with the following unweighted and weighted agreement factors (the maximum parameter shift was 0.005 times its standard uncertainty): R1 = Σ|F o |-|F c | / Σ|F o | = 0.0496 wR 2 ={Σ[w(F o 2 -F c 2 )2] / Σ[w(F o 2 ) 2 } 0.5 =​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​DSC analysis was performed using a TA Instruments Q2500 Discovery Series instrument. The instrument was temperature calibrated using indium. The DSC cell was maintained under a nitrogen purge of approximately 50 mL per minute during each analysis. The sample was placed in a standard crimped aluminum pot and heated from approximately 25°C to 350°C at a rate of 10°C per minute. The DSC thermogram of the crystalline form of compound A is shown in Figure 7. The DSC thermogram of a mixture of the two crystalline forms of compound A is shown in Figure 8.

[0209] Example 7 This embodiment demonstrates the thermogravimetric (TG) characterization of crystalline forms 1 and 2 of compound A (both as pure form 1 and as a mixture of forms 1 and 2) according to embodiments of the present invention.

[0210] TG analysis was performed using a TA Instruments Discovery Q5500 instrument. The instrument balance was calibrated using a Class M weight, and temperature calibration was performed using Alumel. Nitrogen purging was performed at approximately 40 mL / min per minute for the balance and approximately 60 mL / min per minute for the furnace. Each sample was placed in a platinum pan with the container weight removed and heated from approximately 25°C to 350°C at a rate of 10°C per minute. The graph of the thermogravimetric analysis (TGA) of a single crystalline form of compound A is shown in Figure 7. The graph of the TGA of a mixture of two crystalline forms of compound A is shown in Figure 8.

[0211] Example 8 This example illustrates an exemplary method for preparing and characterizing crystalline form 3 of compound A. Approximately 20 mg of amorphous compound A was dissolved in approximately 0.2 mL of 1:1 v:v EtOH / water at ambient temperature (20-25°C). To this solution, approximately 0.06 mL of water was slowly added until a large amount of solid precipitated. The solid was collected by centrifugation filtration through a 0.45 μm nylon membrane filter at 14,000 rpm to obtain crystalline form 3.

[0212] Crystal morphology 3 was characterized by XRPD, DSC, and TGA. XRPD revealed that morphology 3 had a low degree of crystallinity (Figure 9). DSC revealed that morphology 3 had an enthalpy of 23 J / g at 30.2°C. onset It showed a dehydration peak and no clear melting peak after dehydration (Figure 10). TGA showed that form 3 had a weight loss of 3.7% at 115°C (Figure 11).

[0213] Example 9 This example demonstrates an exemplary method for preparing and characterizing crystalline form 4 of compound A. Form 4 arose from the spontaneous crystallization of an oily substance formed by adding 3:7 v:v isopropyl alcohol / water to amorphous compound A. By XRPD, form B had a high degree of crystallinity (Figure 12). After two weeks of storage under ambient conditions (e.g., room temperature), form 4 converted to a disordered substance upon XRPD analysis (Figure 13). By DSC, form 4 showed no endothermic fusion, suggesting that a highly disordered or amorphous substance was likely formed during desolvation (Figure 14). By TGA, form 4 showed broad endothermic fusion of approximately 106°C on DSC (similarly Figure 14).

[0214] Other Embodiments While the present invention is described in relation to its specific embodiments, further modifications are possible, and this application is intended to encompass any modifications, uses, or adaptations, including any deviations from this disclosure that are generally in accordance with the principles of the present invention and that are applicable to the essential features described herein.

[0215] All publications, patents, and patent applications are incorporated herein by reference in the same way as each individual publication, patent, or patent application is specifically and individually indicated to be incorporated by reference in the 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, or form 4.

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

4. The crystal morphology according to claim 3, having at least one peak at a diffraction angle 2θ (°) of 4.4 ± 0.5, 4.6 ± 0.5, or 5.1 ± 0.5, as measured by or calculated from X-ray diffraction by irradiation with Cu Kα X-rays.

5. A mixture of crystalline forms 1 and 2 of compound A: 【Chemistry 2】 or a solvate thereof, the mixture or its solvate having at least one peak at a diffraction angle 2θ (°) of 4.4 ± 0.5, 4.6 ± 0.5, or 4.8 ± 0.5, as measured by or calculated from X-ray diffraction by irradiation with Cu Kα X-rays.

6. A pharmaceutical composition comprising a crystalline form of compound A according to any one of claims 1 to 5 or a solvate thereof, and a pharmaceutically acceptable carrier or excipient.

7. Crystallographic form 1 of compound A or a mixture of crystallographic forms 1 and 2: 【Transformation 3】 or a method for preparing a solvate thereof, comprising: dissolving compound A in a suitable solvent; precipitating crystalline form(s) of compound A by adding a suitable antisolvent; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A.

8. Crystallographic form 1 of compound A or a mixture of crystallographic forms 1 and 2: 【Chemistry 4】 or a method for preparing a solvate thereof, comprising: dissolving compound A in a suitable solvent; precipitating a crystalline form(s) of compound A by evaporation of the suitable solvent; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A.

9. Crystallographic form 1 of compound A or a mixture of crystallographic forms 1 and 2: 【Transformation 5】 or a method for preparing a solvate thereof, comprising: dissolving compound A in a suitable solvent; precipitating the crystalline form(s) of compound A under ambient conditions; isolating the crystalline form(s) of compound A; and drying the crystalline form(s) of compound A.

10. A method for treating cancer in a subject requiring cancer treatment, comprising administering to the subject a therapeutically effective amount of crystalline form 1 or a mixture of crystalline forms 1 and 2 of compound A according to any one of claims 1 to 5, or a solvate thereof, or the pharmaceutical composition according to claim 6.

11. A method for treating a Ras protein-related disorder in a subject requiring treatment for the disorder, comprising administering to the subject a therapeutically effective amount of crystalline form 1 or a mixture of crystalline forms 1 and 2 of compound A according to any one of claims 1 to 5, or a solvate thereof, or the pharmaceutical composition according to claim 6.

12. A method for inhibiting intracellular Ras protein, comprising contacting the cells with an effective amount of crystalline form 1 or a mixture of crystalline forms 1 and 2 of compound A according to any one of claims 1 to 5, or a solvate thereof, or the pharmaceutical composition according to claim 6.

13. The method or use according to any one of claims 10 to 12, further comprising administering additional anticancer therapy.

14. The aforementioned additional anti-cancer therapies are as follows: 【Transformation 6】 The method according to claim 13, wherein the salt is pharmaceutically acceptable or a pharmaceutically acceptable salt thereof.

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