Preparation of compounds and crystalline forms thereof

Abstract

Process for the preparation of the compound (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidin-1-yl)prop-2-en-1-one and crystalline forms thereof.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical chemistry, specifically providing a method for preparing a therapeutic compound and its crystalline form. Background Technology

[0002] TEA domain transcription factors (TEAD proteins) are effectors of oncogenic YAP / TAZ signaling in cancers with dysregulated Hippo tumor suppressor pathway signaling. Interactions between Yesl-associated transcriptional regulators (YAP1, also known as YAP), transcriptional regulator 1 containing the WW domain (WWTR1, also known as TAZ) (a paralog of YAP's transcriptional coactivator), and TEAD proteins have been confirmed both in vitro and in vivo. In both cases, these protein-protein interactions lead to increased TEAD transcriptional activity and increased expression of genes involved in proliferation, survival, angiogenesis, and other cancer markers. In normal tissues, intact Hippo signaling leads to YAP / TAZ phosphorylation, cytoplasmic sequestration, and proteasome degradation via MST1 / 2 and LATS1 / 2 kinases. Somatic mutations in NF2, MST, or LATS inactivate Hippo signaling, stabilizing YAP / TAZ and leading to nuclear translocation, TEAD binding, and activation of TEAD target genes. YAP / TAZ genomic amplification, or gene fusion leading to constitutive activation of YAP / TAZ, is an alternative mechanism for Hippo pathway dysregulation. Furthermore, activation of YAP / TAZ signaling has been described as a mechanism for developing resistance to other targeted cancer therapies, such as EGFR and MEK inhibitors.

[0003] Therefore, inhibitors of TEAD1, 2, 3 and / or 4 have great potential in cancer treatment, both as monotherapy for cancers with dysregulated Hippo pathway signaling and as combination therapy with other targeted drugs.

[0004] The compound (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidone-1-yl)prop-2-en-1-one is described as a covalent inhibitor of YAP / TAZ-TEAD oncoprotein in PCT / US23 / 23957. Summary of the Invention

[0005] It should be understood that the embodiments disclosed herein are not limited to the specific compositions, methods, and experimental conditions described, as such compositions, methods, and conditions can vary. It should also be understood that the terminology used herein is for describing specific embodiments only and is not intended to be limiting.

[0006] This invention provides a method for preparing the compound (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one and its crystalline form.

[0007] Implementation Scheme 1. A method for preparing compound of formula I, Formula I It includes formula 7 The step of reacting the compound with propylene chloride or acrylic anhydride in an organic solvent containing a base reagent.

[0008] Implementation Scheme 2. The method described in Implementation Scheme 1, wherein the reaction conditions for preparing compound I are: 1) The organic solvent is selected from: CH2Cl2, THF, 2-methylTHF, 1,4-dioxane, or acetonitrile; 2) The alkaline reagent is selected from NaHCO3, Na2CO3, K2CO3, KHCO3, Et3N, iPr2NEt, pyridine, or N-methylmorpholine; 3) Reaction time: 1 to 24 hours; 4) Reaction temperature: 0 to 25℃.

[0009] Implementation Scheme 3. The method of Implementation Scheme 1 or 2, wherein the compound of Formula 7 is obtained by deamination protecting group.

[0010] Implementation Scheme 4. The method of Implementation Scheme 3, wherein the amino protecting group is selected from 2-(trimethylsilyl)ethoxycarbonyl (Teoc), 1-methyl-1-(4-biphenyl)ethoxycarbonyl (Bpoc), tert-butoxycarbonyl (BOC), allyloxycarbonyl (Alloc), and benzyloxycarbonyl (Cbz).

[0011] Implementation Scheme 5. The method of Implementation Scheme 3, wherein the compound of Formula 7 is obtained by deamination and dehydration of the compound of Formula 6. Formula 6.

[0012] Implementation Scheme 6. The method of Implementation Scheme 5, wherein the reaction conditions for preparing compound 7 are: 1) The solvent is selected from 2-methyl-THF or 1,4-dioxane; 2) Reaction time: 1 to 10 hours; 3) Reaction temperature: 80 to 100℃; 4) H2SO4 equivalent: 1 to 10 eq.

[0013] Implementation Scheme 7. The method of Implementation Scheme 5, wherein the compound of Formula 6 is obtained by reducing the compound of Formula 5. Formula 5.

[0014] Implementation Scheme 8. The method of Implementation Scheme 7, wherein the reaction conditions for preparing compound of formula 6 are: 1) The solvent is selected from MeOH, EtOH, or THF; 2) The reducing agent is selected from: NaBH4, NaB(CN)H3 or DIBAL-H; 3) Reducing agent equivalent: 0.3 to 1.2 eq.

[0015] Implementation Scheme 9. The method of Implementation Scheme 7, wherein the compound of Formula 5 is obtained by reacting the compound of Formula 3 with a lithium reagent generated from n-BuLi and the compound of Formula 4. Formula 3 Equation 4, In compound formula 4, X is Br or I, and The reaction solvent is selected from: THF, 2-methyl-THF, diethyl ether or 1,4-dioxane.

[0016] Implementation Scheme 10. The method of Implementation Scheme 9, wherein the compound of Formula 3 is obtained by reacting the compound of Formula 1 with the compound of Formula 2. Formula 1 Formula 2 The reaction conditions are as follows: 1) The solvent is selected from: THF, 2-methyl-THF, 1,4-dioxane, DMF or CH2Cl2; 2) Reaction temperature: 0 to 25℃; 3) Reaction time: 1 to 24 hours; 4) The coupling agent is selected from: CDI, HATU, HBTU / HOBT, or EDCI; 5) The base is selected from: Et3N, iPr2NEt, or N-methylmorpholine; 6) Coupling agent equivalent: 1 to 2 eq.

[0017] Implementation Scheme 11. Crystalline form of the compound represented by Formula I: Formula I, The crystalline form is characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 20.4°, 21.1° and 21.7°.

[0018] Implementation Scheme 12. The crystalline form according to Implementation Scheme 11, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 15.2° and 23.0°.

[0019] Implementation Scheme 13. The crystalline form according to Implementation Scheme 12, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 13.6°, 14.0°, 16.5° and 17.0°.

[0020] Implementation Scheme 14. The crystalline form according to any one of Implementation Schemes 11-13, wherein the crystalline form is characterized by being substantially similar to Figure 1 X-ray powder diffraction pattern.

[0021] Implementation Scheme 15. The crystalline form according to any one of Implementation Schemes 11-14, wherein the crystalline form comprises at least 60% monocrystalline form, at least 70% monocrystalline form, at least 80% monocrystalline form, at least 90% monocrystalline form, at least 95% monocrystalline form, or at least 99% monocrystalline form by weight.

[0022] Implementation Scheme 16. The crystalline form according to any one of Implementation Schemes 11-15, wherein the crystalline form has a chemical purity of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% by weight.

[0023] Implementation Scheme 17. The crystalline form according to any one of Implementation Schemes 11-16, wherein the crystalline form is characterized in that the differential scanning calorimetry (DSC) curve includes an endothermic peak of about 87±2°C, preferably about 86.9-88.6°C.

[0024] Implementation Scheme 18. The crystalline form according to Implementation Scheme 17, wherein the crystalline form is characterized by being substantially similar to Figure 2 Differential scanning calorimetry curves.

[0025] Implementation Scheme 19. The crystalline form according to any one of Implementation Schemes 11-18, wherein the crystalline form is anhydrous.

[0026] Implementation Scheme 20. Crystalline form of the compound represented by Formula I: Formula I, The crystalline form is characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 17.3° and 18.3°.

[0027] Implementation Scheme 21. The crystalline form according to Implementation Scheme 20, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) located at 20.4°, 21.1° and 21.7°.

[0028] Implementation Scheme 22. The crystalline form according to Implementation Scheme 20 or 21, wherein the crystalline form is characterized by being substantially similar to Figure 3 The X-ray powder diffraction pattern of the curve.

[0029] Implementation Scheme 23. A method for forming the crystalline form described in any one of Implementation Schemes 11-22, wherein the crystalline form is formed by evaporation or evaporative crystallization, wherein the solvent used in any one of Implementation Schemes 11-19 is selected from THF, ACN, MTBE; and the solvent used in any one of Implementation Schemes 20-22 is selected from acetone, ethyl acetate (EA), methyl ethyl ketone (MEK), and ethanol.

[0030] Implementation Scheme 24. A pharmaceutical composition comprising a pharmaceutically effective amount of any one of the crystalline forms of Implementation Schemes 11-22 and a pharmaceutically acceptable carrier.

[0031] Implementation Scheme 25. A method of treating cancer in a desired subject, comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of Implementation Schemes 11-22 or the pharmaceutical composition of Implementation Scheme 24; or The crystalline form of any one of embodiments 11-22 or the pharmaceutical composition of embodiment 24, for use in treating cancer in a desired subject; or Use of the crystalline form of any one of embodiments 11-22 or the pharmaceutical composition of embodiment 24 in the preparation of a medicament for treating cancer in a subject in need.

[0032] Implementation Scheme 26. According to the method or use of Implementation Scheme 25, wherein the cancer is selected from the group consisting of: bladder cancer, breast cancer, ovarian cancer, pancreatic ductal adenocarcinoma (PDAC), glioblastoma, gastric cancer, cervical cancer, colon cancer, endometrial cancer, head and neck cancer, lung cancer, melanoma, multiple myeloma, leukemia, non-Hodgkin lymphoma, prostate cancer, rectal cancer, malignant melanoma, gastrointestinal cancer, liver cancer, skin cancer, lymphoma, malignant pleural mesothelioma (MPM), kidney cancer, myoma, bone cancer, brain cancer, eye cancer or ocular cancer, rectal cancer, colorectal cancer, cervical cancer, oral cancer. Benign and malignant tumors, gastric cancer, endometrial cancer, testicular cancer, renal cancer, laryngeal cancer, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, neurofibromatosis, tuberous sclerosis, hemangioma and lymphangiogenesis; preferably, the cancers are selected from glioblastoma, gastric cancer, colorectal cancer, pancreatic ductal adenocarcinoma (PDAC) and malignant pleural mesothelioma (MPM).

[0033] Implementation Scheme 27. A method for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample, comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of Implementation Schemes 11-22 or the pharmaceutical composition of Implementation Scheme 24; or The crystalline form of any one of embodiments 11-22 or the pharmaceutical composition of embodiment 24, as a drug for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample; or Use of the crystalline form of any one of embodiments 11-22 or the pharmaceutical composition of embodiment 24 in the preparation of a medicament for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample; Preferably, TEAD1 and 3 are selectively inhibited compared to TEAD2 and 4.

[0034] Implementation scheme 28. Compounds represented by formula 6 or 7, Formula 6, Formula 7.

[0035] definition In this specification and the appended claims, unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” all have plural meanings. Thus, for example, reference to “the method” includes not only one method but also one or more methods and / or steps of the type described herein (steps that will be apparent to those skilled in the art upon reading this disclosure).

[0036] Those skilled in the art will understand the term "about". Whether or not the term "about" is explicitly used, each quantity given herein refers both to the actual value given and to an approximation of that given value that can be reasonably inferred based on the knowledge of those skilled in the art. When "about" is mentioned herein, it includes (and describes) variations with respect to that value or parameter itself. For example, a description of "about X" includes a description of "X". As used herein, the terms "about" and "approximately", unless otherwise stated, when used to refer to temperature, dosage, amount, or weight percentage of an ingredient in a composition / dosage form, refer to a dosage, amount, or weight percentage that is acceptable to those skilled in the art and provides a pharmacological effect equivalent to that produced by the specified dosage, amount, or weight percentage. Specifically, in this context, the terms "about" and "approximately" cover dosage, amount, or weight percentages within the range of 15%, 10%, 5%, 4%, 3%, 2%, 1%, or 0.5% of the specified dosage, amount, or weight percentage.

[0037] As used herein, the term "crystalline" refers to a solid form of Compound I in which the atomic positions exhibit long-range atomic order. The crystalline nature of a solid can be confirmed, for example, by examining an X-ray powder diffraction (XRPD) pattern. If the XRPD pattern shows sharp intensity peaks, the compound is crystalline. The "crystalline" solid form of Compound I refers to a fully crystalline or partially crystalline solid, encompassing solids with at least 80%, 85%, 90%, 95%, and 99% crystalline content by weight. A crystalline solid is a molecularly ordered arrangement with a distinguishable lattice or unit cell, and therefore possesses definable long-range order. In some embodiments, "substantially crystalline" refers to a solid form of Compound I with at least 80% crystalline content by weight. In other embodiments, "substantially crystalline" refers to a solid form with at least 90% crystalline content by weight.

[0038] As used herein, "chemical purity" refers to the degree to which the disclosed form does not contain substances with different chemical structures. The chemical purity of a compound in the disclosed crystalline form is defined as the weight of the compound divided by the sum of the weight of the compound and the weight of substances / impurities with different chemical structures, multiplied by 100%, i.e., weight percentage. In one embodiment, the compound in the disclosed crystalline form has a weight chemical purity of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%.

[0039] The term "hydrate" refers to a crystalline compound I in which stoichiometric or non-stoichiometric amounts of water are bound into the crystal structure. A hydrate is a solvate in which water is the solvent bound into the crystal structure. When used to describe a compound, the term "anhydrous" means that the crystal structure contains essentially no bound solvent, for example, less than 0.1% by weight according to Karl Fischer analysis. Anhydrous compounds are referred to herein as "anhydrous compounds".

[0040] The term "amorphous" refers to a solid existing in a non-crystalline state or form. Amorphous solids are disordered arrangements of molecules and therefore lack a distinguishable lattice or unit cell, as well as definable long-range order. The solid-state order of a solid can be determined using standard techniques known in the art, such as X-ray powder diffraction (XRPD) or differential scanning calorimetry (DSC). Amorphous solids can also be distinguished from crystalline solids, for example, by observing birefringence using a polarized light microscope.

[0041] A single crystalline form of compound I refers to (R, E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one existing in a single crystalline form or multiple crystalline forms, wherein each crystal has the same crystalline form (i.e., crystalline form I or II). When a crystalline form is defined as a specific percentage of a particular single crystalline form of the compound, the remainder consists of an amorphous form and / or other crystalline forms besides the specified specific crystalline form. In one embodiment, the crystalline form is at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% by weight of the single crystalline form. The weight percentage of a particular crystalline form is determined by dividing the weight of the particular crystalline form by the sum of (the weight of the particular crystalline form + the weight of other crystalline forms present + the weight of the amorphous form present), and then multiplying by 100%.

[0042] The 2-theta values ​​of the X-ray powder diffraction patterns for the crystal types described herein may vary slightly due to differences in instrumentation and sample preparation and batch-to-batch variations. Therefore, unless otherwise defined, the XRPD patterns and / or 2-theta peaks described herein should not be considered absolute values ​​and may have a deviation of ±0.2 degrees. The 2-theta values ​​provided herein were obtained using Cu K-alpha radiation.

[0043] Temperature values ​​(e.g., DSC peak temperature and DSC onset temperature) in this document may vary slightly due to differences in instruments, sample preparation, heating rates during experiments, batch-to-batch variations in materials, and other environmental factors. Therefore, unless otherwise defined, temperature values ​​described herein should not be considered absolute values ​​and may have a deviation of ±5 degrees. “Substantially identical XRPD patterns” or “X-ray powder diffraction patterns substantially similar to…” (for a particular figure) means that, for comparative purposes, at least 90% of the peaks shown in the figure are present. Furthermore, it should be understood that, for comparative purposes, the 2-theta (2θ) peak positions are allowed to deviate from the positions shown, for example, ±0.2°. It should be understood that when “characterized by X-ray powder diffraction peaks at 2θ angles (±0.2°)” is followed by a series of 2θ peak positions, this ±0.2° deviation applies to each of the listed peak positions.

[0044] Pharmaceutical preparations and methods of treatment and administration In this article, the terms "treatment" and "therapeutic method" are used interchangeably, referring to both 1) therapeutic treatments or measures aimed at curing, alleviating, reducing symptoms, and / or stopping the progression of a diagnosed pathological condition, disease, or disorder, and 2) preventive measures. Those requiring treatment include both individuals who already have a specific disease or disorder and individuals who may eventually develop that disorder (i.e., the population requiring preventive measures).

[0045] In some or any embodiment, “treating” for any condition or disorder means improving the condition or disorder present in the subject, including preventative improvement. In another embodiment, “treatment” includes improving at least one physical parameter that may not be perceptible to the subject. In yet another embodiment, “treatment” includes modulating the condition or disorder, whether through physical modulation (e.g., stabilizing perceptible symptoms), physiological modulation (e.g., stabilizing physical parameters), or both. In yet another embodiment, “treatment” includes delaying the onset of the condition or disorder. In yet another embodiment, “treatment” includes reducing or eliminating the condition (e.g., cancer) or one or more parameters of the condition (e.g., cancer) (e.g., tumor size or tumor growth rate), or delaying the progression of the condition (e.g., cancer) or one or more parameters of the condition (e.g., cancer) (e.g., tumor size or tumor growth rate), or reducing the severity of the condition (e.g., cancer) or one or more parameters of the condition (e.g., cancer) (e.g., tumor size or tumor growth rate). In yet another embodiment, “treatment” includes the preventative administration of the compounds described herein. In some implementations, “treatment” of a disease includes: (1) suppressing the disease, i.e. preventing (i.e. stabilizing) or reducing the development of the disease or its clinical symptoms; or (2) alleviating the disease, even if the disease or its clinical symptoms subside.

[0046] As used herein, the term "subject" refers to any individual or patient who receives the methods described herein. Typically, a subject is a human being, although those skilled in the art will understand that a subject can also be an animal.

[0047] The terms “therapeutic effective amount,” “effective dose,” “therapeutic effective dose,” “effective amount,” and similar terms refer to an amount of a test compound or composition sufficient to elicit the desired biological or medical response in a tissue, system, animal, individual, or human by administration of the compound.

[0048] This document also discloses pharmaceutical compositions comprising the crystal forms of the compounds described herein. The term "pharmaceuticalally acceptable carrier" refers to a non-toxic carrier that can be administered to a patient in conjunction with the compounds disclosed herein without impairing the pharmacological activity of the compounds.

[0049] The crystal forms of the compounds disclosed herein can be used in a conventional manner to control the diseases described herein (including, but not limited to, cancer). Such treatment methods, along with their dosage levels and requirements, can be selected by those skilled in the art based on existing methods and techniques. For example, the compounds of this disclosure can be combined with pharmaceutically acceptable adjuvants and administered to patients with cancer in a pharmaceutically acceptable manner and at doses effective in treating cancer.

[0050] Alternatively, the polymorphs of the compounds disclosed herein may be used in compositions and methods to treat the diseases described herein (including, but not limited to, cancer) or protect individuals from such diseases for a prolonged period of time. These polymorphs may be used alone or in combination with other compounds of this disclosure, consistent with the conventional use of such compounds in pharmaceutical compositions. For example, the polymorphs of the compounds disclosed herein may be combined with pharmaceutically acceptable adjuvants commonly used in vaccines and administered in a preventatively effective amount to protect individuals from the diseases described herein (including, but not limited to, cancer) for a prolonged period of time.

[0051] As used herein, the terms “combination,” “in combination,” and related terms refer to the simultaneous or sequential administration of therapeutic agents according to this disclosure. For example, the crystalline form of the compound may be administered simultaneously or sequentially with another therapeutic agent, either individually in unit dosage forms or in combination in a single unit dosage form. Therefore, this disclosure provides a unit dosage form comprising the crystalline form of the compound, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or excipient. When a patient or individual is simultaneously exposed to two or more drugs, these drugs are generally considered to be administered in “combination.” In many embodiments, two or more drugs are considered to be administered in “combination” when a patient or individual simultaneously presents therapeutically significant drug levels in a specific target tissue or sample (e.g., brain, serum, etc.).

[0052] When the crystalline forms of the compounds disclosed herein are used in combination with other drugs, they may be administered sequentially or simultaneously. Alternatively, the pharmaceutical or preventative compositions of this disclosure comprise a combination of the crystalline forms of the compounds described herein with another therapeutic or preventative drug. Other therapeutic drugs commonly used to treat a particular disease or condition may be referred to as "drugs for the treated disease or condition".

[0053] The compounds used in the compositions and methods of this disclosure can also be modified by adding appropriate functional groups to enhance their selective biological properties. Such modifications are known in the art and include increasing their biopermeability in specific biological systems (e.g., blood, lymphatic system, or central nervous system), improving oral bioavailability, improving solubility to allow for injection administration, altering metabolism, and / or changing excretion rates.

[0054] According to one embodiment, the compositions of this disclosure are formulated for drug administration to a subject or patient (e.g., a mammal, preferably a human). Such pharmaceutical compositions are intended to improve, treat, or prevent any of the diseases described herein, including but not limited to cancer.

[0055] The formulations disclosed herein are typically administered as pharmaceutical compositions comprising an active therapeutic agent, i.e., and various other pharmaceutically acceptable ingredients. See Remington Pharmaceutical Sciences (15th ed., Mack Publishers, Easton, PA, 1980). Preferred dosage forms depend on the intended route of administration and therapeutic use. Depending on the desired formulation, these compositions may also contain pharmaceutically acceptable nontoxic carriers or diluents commonly used in the formulation of pharmaceutical compositions for animal or human use. When selecting a diluent, it should be ensured that it does not affect the biological activity of the composition. Examples of such diluents include distilled water, physiological phosphate-buffered saline, Ringer's solution, glucose solution, and Hanks' solution. Furthermore, the pharmaceutical compositions or formulations may also contain other carriers, adjuvants, or nontoxic, non-therapeutic, non-immunogenic stabilizers, etc.

[0056] In some embodiments, this disclosure provides pharmaceutically acceptable compositions comprising a therapeutically effective amount of one or more of the said compound crystal forms, formulated together with one or more pharmaceutically acceptable carriers (additives) and / or diluents, for the treatment of the diseases described herein, including but not limited to cancer. While the said compound crystal forms may be administered alone, it is preferred to administer the compound crystal forms as pharmaceutical formulations (compositions) described herein. The compound crystal forms may be formulated in any convenient manner, analogous to other pharmaceuticals, for human or veterinary medicine.

[0057] The compounds described herein, when used as pharmaceuticals in humans and animals in crystalline form, can be administered alone or as pharmaceutical compositions, for example, in combination with a pharmaceutically acceptable carrier at a concentration of 0.1% to 99.5% of the active ingredient. In some embodiments, the concentration of the active ingredient may be 0.5% to 90%.

[0058] The formulations described herein can be administered orally, nasally (e.g., spray), parenterally, vaginally, intracerebrospinally, rectally, or topically (e.g., powder, ointment, or drops, including buccal and sublingual administration). Naturally, the dosage form should be suitable for the corresponding route of administration. For example, they can be formulated as tablets or capsules for administration via injection, inhalation, eye wash, ointment, suppository, etc.; topically (wash or ointment); and rectally (suppository). Oral administration is the preferred route of administration.

[0059] Regardless of the route of administration chosen, the crystalline form of the compounds described herein (which may be in a suitable hydrated form) and / or the pharmaceutical compositions disclosed herein can be formulated into pharmaceutically acceptable dosage forms using conventional methods known to those skilled in the art.

[0060] The actual dosage level of the active ingredient in the pharmaceutical composition disclosed herein can be adjusted to obtain an effective dosage of the active ingredient that can achieve the desired therapeutic effect for a specific patient, composition, and route of administration.

[0061] The crystalline form of the compound described in this disclosure can be administered in combination with one or more other therapeutic agents. The terms "combination therapy," "in combination with," and similar expressions refer to the simultaneous use of more than one drug or treatment method to enhance efficacy. For example, the TEAD1, 2, 3, and / or 4 inhibitors of this disclosure can be used in combination with other drugs or therapies used to treat cancer. In various respects, the crystalline form of the compound can be administered before, simultaneously with, or after chemotherapy.

[0062] In another embodiment, this disclosure relates to the use of the crystal form of a compound of formula I and / or a pharmaceutically acceptable salt thereof in the treatment of any cancer indication in which YAP is localized within the nucleus of tumor cells, including but not limited to lung cancer, thyroid cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, esophageal cancer, liver cancer, breast cancer, and skin cancer.

[0063] In another embodiment, the disclosed compound crystal form or pharmaceutical composition thereof can disrupt the interaction between YAP and TEAD1, 2, 3, and / or 4. In some embodiments, the disclosed compound crystal form or pharmaceutical composition thereof can prevent YAP from binding to TEAD1, 2, 3, and / or 4. In some embodiments, the disclosed compound crystal form or pharmaceutical composition thereof can competitively bind to TEAD1, 2, 3, and / or 4 with YAP. In some embodiments, the disclosed compound crystal form or pharmaceutical composition thereof can bind to TEAD1, 2, 3, and / or 4. In some embodiments, the disclosed compound or pharmaceutical composition thereof can bind to TEAD1 and 3. In some embodiments, the disclosed compound or pharmaceutical composition thereof can selectively bind to TEAD1 and 3 without binding to other TEAD isoforms.

[0064] The term "pharmaceutically acceptable" means that the carrier, diluent, or excipient must be compatible with other components in the formulation and harmless to the recipient. For example, the carrier, diluent, excipient, or a combination thereof can be administered to a subject together with the TEAD1, 2, 3, and / or 4 inhibitors of this disclosure without causing any adverse biological effects or adverse interactions with the TEAD1, 2, 3, and / or 4 inhibitors contained in the pharmaceutical composition. Attached Figure Description

[0065] Figure 1 XRPD diffraction pattern of crystalline form I Figure 2 DSC curve of crystalline form I Figure 3XRPD diffraction pattern of acetone in crystalline form II Example Although other methods and materials similar to or equivalent to those described herein may be used in carrying out or testing the present invention, it should be understood that such modifications and variations are within the scope and spirit of this patent. Preferred methods and materials will be described in detail below.

[0066] Example 1 Preparation method of compound I ((R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolid-1-yl)prop-2-en-1-one) Step 1: Synthesis of (R)-3-(2-(methoxy(methyl)amino)-2-oxoethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3 THF (21.0 L, 7.00 v / w) was added to the reactor (50.0 L) at 15–25 °C, followed by (R)-2-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)acetic acid 1 (3.0 kg, 13.1 mol, 1.00 equivalent), then TEA (1.72 kg, 17.0 mol, 1.30 equivalent) and CDI (2.33 kg, 14.4 mol, 1.10 equivalent) at 15–25 °C. The mixture was stirred at 15–25 °C for 2 hours. Then, N,O-dimethylhydroxylamine hydrochloride 2 (1.40 kg, 14.4 mol, 1.10 equivalent) was added to the reactor at 15–25 °C. The resulting solution was stirred at 15–25 °C for 15 hours, and HPLC showed that the reaction was complete. The reaction solution was filtered, and the wet filter cake was washed with EtOAc (6.00 L, 2.00 v / w). The filtrate was concentrated to 2.00 to 3.00 V, and EtOAc (15.0 L, 5.00 v / w) was added at 15 to 25 °C. The organic phase was washed with aqueous HCl solution (1 N, 1.50 L, 5.00 v / w) (pH 3–4). The organic phase was separated and washed with NaCl solution (1.50 L, 15.0% w / w, 5.00 v / w). The organic phase was concentrated to give (R)-3-(2-(methoxy(methyl)amino)-2-oxoethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3 (3.03 kg, 85% yield), as a brown oil with a purity of 94.5%.

[0067] MS (ESI, m / z), 217.1 [M- t Bu+1] + . 1H NMR (400 MHz, CDCl3) δ (ppm) 3.68(d, 3H), 3.64 – 3.58 (m, 1H), 3.49 – 3.41 (m, 1H), 3.34 – 3.25 (m, 1H), 3.19(d, 3H), 2.98 – 2.92 (m, 1H), 2.67 – 2.58 (m, 1H), 2.51 (m, 2H), 2.14 – 2.04(m, 1H), 1.62 – 1.55 (m, 1H), 1.49 (d, 9H).

[0068] Synthesis of step 2(R)-3-(2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 5 THF (4.5 L, 3.00 v / w) was added to a reactor (50.0 L) at 15–25 °C, and the solution was cooled to -60–70 °C. n-BuLi (5.51 L, 2.50 M, 13.8 mol, 2.50 equivalents) was added dropwise to the reactor at -60–70 °C, followed by a THF solution (1.50 L, 1.00 v / w) of 1-bromo-4-(trifluoromethyl)benzene (3.10 kg, 13.8 mol, 2.50 equivalents)⁴. After the additions were complete, the reaction mixture was stirred at -60–70 °C for 1 hour. A solution of (R)-3-(2-(methoxy(methyl)amino)-2-oxoethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 3 (1.50 kg, 5.51 mol, 1.00 equivalent) in THF (1.50 L, 1.00 v / w) was added dropwise to the reactor at -60 to -70 °C. The mixture was stirred at -60 to -70 °C for 2 h. HPLC showed the reaction was complete. An aqueous solution of HCl (2.00 N, 7.50 L, 5.00 v / w) was added dropwise to the reactor at -60 to -70 °C. The reaction was then gradually heated to 0 to 10 °C. EtOAc (10.5 L, 7.00 v / w) was added to the mixture. The organic phase was separated, washed with an aqueous solution of NaHCO3 (7.50 L, 5.00 v / w), and then washed with an aqueous solution of NaCl (7.50 L, 5.00 v / w). The organic phase was concentrated under reduced pressure at 40–50 °C. Heptane (10.5 L, 7.00 v / w) was added to the reactor, and the suspension was concentrated under reduced pressure at 40–50 °C until the volume was 2.00–3.00 v. Then, heptane (7.50 L, 5.00 v / w) was added back to the reactor. The suspension was cooled to 0–10 °C and stirred for 2 hours. The suspension was filtered, and the wet filter cake was washed with heptane (1.50 L, 1.00 v / w). The filter cake was dried in an oven at 40–50 °C. 5 (1.68 kg, 82.0% yield) of (R)-3-(2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester was given as a grayish-white solid with 100% purity.

[0069] MS (ESI, m / z), 302.1[m-tBu+1]+. 1H NMR (400 MHz, CDCl3) δ (ppm) 8.08–8.04 (m, 2H), 7.76–7.73 (m, 2H), 3.72–3.66 (m, 1H), 3.52–3.45 (m, 1H), 3.38–3.29 (m, 1H) , 3.05 (d, J=3.2Hz, 2H), 2.99–2.85 (m, 1H), 2.82–2.70 (m, 1H), 2.21–2.11 (m, 1H), 1.67–1.52 (m, 1H), 1.46 (s, 9H).

[0070] Step 3: Synthesis of (3R)-3-(2-hydroxy-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 6 EtOH (17.5 L, 7.00 v / w) was added to the reactor (50.0 L) at 15–25 °C, followed by (R)-3-(2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 5 (2.50 kg, 7.00 mol, 1.00 equivalent). The suspension was cooled to 0–10 °C, and NaBH4 (79.5 g, 2.10 mol, 0.30 equivalent) was added to the reactor at 0–10 °C. The resulting mixture was stirred at 0–10 °C for 1 hour, and the reaction was complete as indicated by HPLC. An aqueous solution of HCl (1.50 L, 0.60 v / w) was added dropwise to the reactor at 0–10 °C. The suspension was filtered, and the wet filter cake was washed with EtOAc (5.00 L, 2.00 v / w). Water (12.5 L, 5.00 v / w) was added to the above mixture, and the solution was concentrated to 5.00 to 6.00 v. EtOAc (17.5 L, 7.00 v / w) was added. The organic phase was separated and washed with an aqueous NaCl solution (12.5 L, 5.00 v / w). The organic phase was concentrated under reduced pressure at 40 to 50 °C. Heptane (12.5 L, 5.00 v / w) was added to the reactor. The suspension was stirred at 15 to 25 °C for 2 hours and then filtered. The wet filter cake was washed with heptane (2.50 L, 1.00 v / w) and dried in an oven at 40 to 50 °C. (3R)-3-(2-hydroxy-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 6 (2.25 kg, 90% yield) was given as an orange solid with a purity of 100%.

[0071] MS (ESI, m / z), 382.1[m+Na]+. 1H NMR (400 MHz, CDCl3) δ (ppm) 7.62-7.60 (m, 2H), 7.49-7.47 (m, 2H), 4.78-4.77 (m, 1H), 3.64- 3.40 (m, 2H), 3.22-3.20 (m, 1H), 2.97-2.59 (m, 2H), 2.02-1.50 (m, 5H), 1.43 (s, 9H).

[0072] Step 4: Synthesis of (R,E)-3-(4-(trifluoromethyl)styryl)pyrrolidine 7 1,4-Dioxane (17.5 L, 7.00 v / w) was added to the reactor at 15–25 °C, followed by concentrated H₂SO₄ (4.09 kg, 41.8 mol, 6.00 equivalent) at the same temperature. Then, (3R)-3-(2-hydroxy-2-(4-(trifluoromethyl)phenyl)ethyl)pyrrolidine-1-carboxylic acid tert-butyl ester 6 (2.50 kg, 6.96 mol, 1.00 equivalent) was added to the reactor at 15–25 °C. The resulting suspension was stirred at 15–25 °C for 2 hours, then heated to 95–105 °C. The reaction solution was stirred at 100 °C for another 3 hours, and HPLC indicated completion of the reaction. The solution was cooled to 15–25 °C and then poured into water (12.5 L, 5.00 v / w) at the same temperature. The solution was concentrated to 5.00 to 7.00 V and cooled to 0 to 10 °C. At 0 to 10 °C, the mixture was adjusted to pH approximately 11 to 12 with an aqueous NaOH solution (40% w / w). The suspension was filtered, and the solid was washed with CH₂Cl₂ / MeOH (7.50 L, 3.00 v / w, 6 / 1). CH₂Cl₂ / MeOH (17.50 L, 3.00 v / w, 6 / 1) was added to the filtrate. The organic layer was separated. The aqueous layer was extracted with CH₂Cl₂ / MeOH (17.50 L, 3.00 v / w, 6 / 1). The organic phases were combined and washed with water (12.5 L, 5.00 v / w). The organic phase was concentrated under reduced pressure to give (R,E)-3-(4-(trifluoromethyl)styryl)pyrrolidine 7 (1.53 kg, 91.5% yield), as a brown solid with a purity of 91.5%.

[0073] MS (ESI, m / z), 242.1[m+1]+. 1H NMR (400 MHz, CDCl3) δ (ppm) 7.47-7.45 (m, 2H), 7.36-7.34 (m, 2H), 6.39-6.32 (m, 1H), 6.23-6.17 (m, 1H), 3.15-3.10 (m, 1H), 2.98-2.96 (m, 2H), 2.83-2.65 (m, 3H), 2.02-1.97 (m, 1H), 1.61-1.55 (m, 1H).

[0074] Step 5 (R, E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one compound of formula I Acetonitrile (17.5 L, 7.00 v / w) was added to the reactor at 10–20 °C, followed by (R,E)-3-(4-(trifluoromethyl)styryl)pyrrolidine 7 (2.50 kg, 10.4 mol, 1.00 equivalent) and NaHCO3 (0.81 kg, 10.4 mol, 1.00 equivalent) at 10–20 °C. The suspension was cooled to 0–10 °C, and acrylonitrile-2-enoyl chloride (0.94 kg, 11.2 mol, 1.08 equivalent) was added dropwise to the reactor at 0–10 °C. The mixture was stirred at 0–10 °C for 3 hours, and the reaction was complete as indicated by HPLC. The solution was filtered, and the wet filter cake was washed with EtOAc (5.00 L, 2.00 v / w). The filtrate was concentrated to 1.00–2.00 V, and then EtOAc (12.5 L, 5.00 V / w) was added. The organic solution was washed with an aqueous NaHCO3 solution (7.50 L, 3.00 v / w). The organic phase was concentrated to 1.00 to 2.00 V, and then MTBE (12.5 L, 5.00 v / w) was added. The solution was concentrated to 1.00 to 2.00 V, and another portion of MTBE (12.5 L, 5.00 v / w) was added. The solution was concentrated to 2.00 to 3.00 V and cooled to -10 to 0 °C. The mixture was stirred at -10 to 0 °C for 4 hours. The suspension was filtered, and the wet filter cake was washed with MTBE (2.50 L, 1.00 v / w). The filter cake was dried in a vacuum oven at 30 to 40 °C for 8 hours. (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one (3.50 kg, yield 64.9%) was obtained as a grayish-white crystalline solid with a purity of 98.3% and 100% ee.

[0075] MS (ESI, m / z), 296.10 [M+1] + . 1H NMR (400 MHz, CDCl3) δ (ppm) 7.58 -7.55 (m, 2H), 7.45 - 7.43 (m, 2H), 6.55 - 6.41 (m, 3H), 6.27 (m, 1H), 5.71 -5.68 (m, 1H), 3.89 - 3. 35 (m, 4H), 3.12 -3.00 (m, 1H), 2.23 - 1.81 (m, 2H); 19 F NMR (376 MHz, CDCl3) δ (ppm) -62.477, -62.492. Melting point, 87.8-88.6℃.

[0076] The product of Example 1 was characterized by XRPD (Cu, k-Alpha λ = 1.54184 Å). The obtained XRPD spectrum is shown below. Figure 1 Its DSC curve is shown in Figure 2 The crystalline form prepared in Example 1 was named Crystal Form I, characterized by X-ray powder diffraction peaks at 2θ angles (±0.2°) of 20.4°, 21.1°, 21.7°, 15.2°, 23.0°, 13.6°, 14.0°, 16.5°, and 17.0°. Furthermore, the melting point onset and termination temperatures of the product were measured three times, and the results are shown in Table 1. The peak positions of the XRPD spectra are shown in Table 2.

[0077] Table 1. Starting and ending temperatures of the melting point of compound I in Example 1

[0078] Table 2. XRPD peaks of compound I in Example 1

[0079] Example 2 Another method for preparing compound I ((R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolid-1-yl)prop-2-en-1-one) Prepare the racemic product of 1-{3-[(E)-2-[4-((trifluoromethyl)styryl)pyrrolid-1-yl)prop-2-en-1-one} according to PCT / US23 / 23957, and estimate its MS (ESI) mass. C 16 H 16F3NO, 295.10 m / z, measured value, 296.10 [M+H]+; 1H NMR (400 MHz, DMSO-d6) δ 7.74 - 7.62 (m, 4H), 6.69 - 6.45 (m, 3H), 6.18 - 6.09 (m, 1H), 5.74 - 5.62 (m, 1H), 3.89 - 3.51 (m, 3H), 3.23 -2.96 (m, 2H), 2.18 - 2.01 (m, 1H), 1.95 - 1.69 (m, 1H); 19F NMR (376 MHz, DMSO-d6) δ (ppm): -60.83.

[0080] According to patent application number PCT / US23 / 23957, (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one was obtained by chiral separation of the above racemic compound.

[0081] Compound I The racemic products were separated by SFC (column: CHIRAL ART Cellulose-SB, 2). 25 cm, 5 μm. Mobile phase A: Hex (0.5% 2M NH3-MeOH)-HPLC, mobile phase B: IPA-HPLC; flow rate: 20 mL / min; gradient: 15% B to 15% B 21 min; wavelength: 220 / 254 nm; RT1 (min): 14.491 (first isomer); sample solvent: EtOH-HPLC; injection volume: 0.4 mL; number of runs: 2) A stereoisomer compound I was obtained, designated as the R configuration (second eluting isomer), (R,E)-1-(3-(4-(trifluoromethyl)styryl)pyrrolidine-1-yl)prop-2-en-1-one, as a white solid (3.8 mg). C 16 H 16 The calculated MS (ESI) mass of F3NO is 295.10 m / z, while the measured value is 296.10 [M+H]. + ). 1 H NMR (400 MHz, DMSO- d6) δ 7.74 - 7.62 (m, 4H), 6.69- 6.45 (m, 3H), 6.18 - 6.09 (m, 1H), 5.74 - 5.62 (m, 1H), 3.89 - 3.51 (m,3H), 3.23 - 2.96 (m, 2H), 2.18 - 2.01 (m, 1H), 1.95 - 1.69 (m, 1H). 19 F NMR (376 MHz, DMSO- d 6 δ (ppm): -60.82.

[0082] Example 3 Biological activity assay of compound I The ability of compound I to inhibit TEAD-dependent transcription was evaluated in the MCF7-TEAD-Luc reporter assay (BPS #60618). For data analysis, dose-dependent curves were plotted as “% inhibition” (% of DMSO-treated control wells), and IC50 values ​​were obtained by fitting a four-parameter model in GraphPad Prism. TEAD1-Luc MCF7: mean IC50 value ≤10 nM; TEAD1-Luc MCF7: maximum inhibition rate ≥75%.

[0083] The potency and efficacy of compound I in inhibiting cell proliferation and viability were evaluated in multiple cell lines. The mean IC50 was measured in NCI-226 cells. 50 Values ​​less than 100 nM, with maximum average inhibition rates ranging from 50% to 75%. The average IC50 value measured in H2052 cells... 50 The value is less than 100 nM, and the maximum average inhibition rate is 50%-75%. In the selectivity test of TEAD, the 3 μm compound I inhibited TEAD1, TEAD2, TEAD3 and TEAD4 by 94%, 57%, 98% and 52%, respectively.

[0084] Example 4 Example 2: Preparation and characterization of the crystalline form of compound I Referring to Table 3, 5 mg of compound I prepared in Example 2 was dissolved in a specific solvent, and the solid obtained by evaporation and recrystallization was characterized by XRPD (Cu, k-Alpha λ = 1.54184 Å). The product prepared from THF (tetrahydrofuran), ACN (acetonitrile), and MTBE (methyl tert-butyl ether) was characterized as crystalline form I. The product prepared from acetone, EA (ethyl acetate), MEK (methyl ethyl ketone), and ethanol had a different XRPD powder diffraction pattern than crystalline form I and was named crystalline form II (see Table 3). Figure 3 (See Table 4). Crystal form II is characterized by X-ray powder diffraction peaks at 2θ angles (±0.2°) of 17.3°, 18.3°, 20.4°, 21.1°, and 21.7°. Compared with compound I prepared in Example 2, crystal form I and crystal form II have different XRPD patterns.

[0085] Table 3 Characterization of substances prepared by evaporation crystallization method

[0086] Table 4. XRPD peaks of crystal form II prepared with acetone in Example 2.

Claims

1. A method for preparing compound of formula I, Equation I It includes formula 7 The step of reacting the compound with propylene chloride or acrylic anhydride in an organic solvent containing a base reagent.

2. The method of claim 1, wherein the reaction conditions for preparing compound I are: 1) The organic solvent is selected from: CH2Cl2, THF, 2-methylTHF, 1,4-dioxane, or acetonitrile; 2) The alkaline reagent is selected from NaHCO3, Na2CO3, K2CO3, KHCO3, Et3N, iPr2NEt, pyridine, or N-methylmorpholine; 3) Reaction time: 1 to 24 hours; 4) Reaction temperature: 0 to 25℃.

3. The method of claim 1 or 2, wherein the compound of formula 7 is obtained by deamination protecting group.

4. The method of claim 3, wherein the amino protecting group is selected from 2-(trimethylsilyl)ethoxycarbonyl (Teoc), 1-methyl-1-(4-biphenyl)ethoxycarbonyl (Bpoc), tert-butoxycarbonyl (BOC), allyloxycarbonyl (Alloc), and benzyloxycarbonyl (Cbz).

5. The method of claim 3, wherein the compound of formula 7 is obtained by deamination and dehydration of the compound of formula 6. Formula 6.

6. The method of claim 5, wherein the reaction conditions for preparing the compound of formula 7 are: 1) The solvent is selected from 2-methyl-THF or 1,4-dioxane; 2) Reaction time: 1 to 10 hours; 3) Reaction temperature: 80 to 100℃; 4) H2SO4 equivalent: 1 to 10 eq.

7. The method of claim 5, wherein the compound of formula 6 is obtained by reducing the compound of formula 5. Formula 5.

8. The method of claim 7, wherein the reaction conditions for preparing the compound of formula 6 are: 1) The solvent is selected from MeOH, EtOH, or THF; 2) The reducing agent is selected from: NaBH4, NaB(CN)H3 or DIBAL-H; 3) Reducing agent equivalent: 0.3 to 1.2 eq.

9. The method of claim 7, wherein the compound of formula 5 is obtained by reacting the compound of formula 3 with a lithium reagent generated from n-BuLi and the compound of formula 4. Formula 3 Equation 4, In compound formula 4, X is Br or I, and The reaction solvent is selected from: THF, 2-methyl-THF, diethyl ether or 1,4-dioxane.

10. The method of claim 9, wherein the compound of formula 3 is obtained by reacting the compound of formula 1 with the compound of formula 2. Formula 1 Formula 2 The reaction conditions are as follows: 1) The solvent is selected from: THF, 2-methyl-THF, 1,4-dioxane, DMF or CH2Cl2; 2) Reaction temperature: 0 to 25℃; 3) Reaction time: 1 to 24 hours; 4) The coupling agent is selected from: CDI, HATU, HBTU / HOBT, or EDCI; 5) The base is selected from: Et3N, iPr2NEt, or N-methylmorpholine; 6) Coupling agent equivalent: 1 to 2 eq.

11. Crystalline form of the compound represented by Formula I: Equation I, The crystalline form is characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 20.4°, 21.1° and 21.7°.

12. The crystalline form according to claim 11, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 15.2° and 23.0°.

13. The crystalline form according to claim 12, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 13.6°, 14.0°, 16.5° and 17.0°.

14. The crystalline form according to any one of claims 11-13, wherein the crystalline form is characterized by being substantially similar to the X-ray powder diffraction pattern of FIG1.

15. The crystalline form according to any one of claims 11-14, wherein the crystalline form comprises at least 60% monocrystalline form, at least 70% monocrystalline form, at least 80% monocrystalline form, at least 90% monocrystalline form, at least 95% monocrystalline form, or at least 99% monocrystalline form by weight.

16. The crystalline form according to any one of claims 11-15, wherein the crystalline form has a chemical purity of at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% by weight.

17. The crystalline form according to any one of claims 11-16, wherein the crystalline form is characterized in that the differential scanning calorimetry (DSC) curve includes an endothermic peak at about 87±2°C, preferably about 86.9-88.6°C.

18. The crystalline form according to claim 17, wherein the crystalline form is characterized by being substantially similar to the differential scanning calorimetry curve of Figure 2.

19. The crystalline form according to any one of claims 11-18, wherein the crystalline form is anhydrous.

20. Crystalline form of the compound represented by Formula I: Equation I, The crystalline form is characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) at 17.3° and 18.3°.

21. The crystalline form according to claim 20, wherein the crystalline form is further characterized by having X-ray powder diffraction peaks at 2θ angles (±0.2°) located at 20.4°, 21.1° and 21.7°.

22. The crystalline form according to claim 20 or 21, wherein the crystalline form is characterized by an X-ray powder diffraction pattern substantially similar to the curve in Figure 3.

23. A method for forming the crystalline form according to any one of claims 11-22, wherein the crystalline form is formed by evaporation or evaporative crystallization, wherein the solvent used in any one of claims 11-19 is selected from THF, ACN, and MTBE; and the solvent used in any one of claims 20-22 is selected from acetone, ethyl acetate (EA), methyl ethyl ketone (MEK), and ethanol.

24. A pharmaceutical composition comprising a pharmaceutically effective amount of the crystalline form of any one of claims 11-22 and a pharmaceutically acceptable carrier.

25. A method of treating cancer in a desired subject, comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24; or The crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24, for use in treating cancer in a desired subject; or Use of the crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating cancer in a subject in need.

26. The method or use according to claim 25, wherein the cancer is selected from the group consisting of: bladder cancer, breast cancer, ovarian cancer, pancreatic ductal adenocarcinoma (PDAC), glioblastoma, gastric cancer, cervical cancer, colon cancer, endometrial cancer, head and neck cancer, lung cancer, melanoma, multiple myeloma, leukemia, non-Hodgkin's lymphoma, prostate cancer, rectal cancer, malignant melanoma, gastrointestinal cancer, liver cancer, skin cancer, lymphoma, malignant pleural mesothelioma (MPM), kidney cancer, myoma, bone cancer, brain cancer, eye cancer or ocular cancer, rectal cancer, colorectal cancer, cervical cancer, oral cancer. Benign and malignant tumors, gastric cancer, endometrial cancer, testicular cancer, renal cancer, laryngeal cancer, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, neurofibromatosis, tuberous sclerosis, hemangioma and lymphangiogenesis; preferably, the cancers are selected from glioblastoma, gastric cancer, colorectal cancer, pancreatic ductal adenocarcinoma (PDAC) and malignant pleural mesothelioma (MPM).

27. A method for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample, comprising administering to the subject a therapeutically effective amount of the crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24; or The crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24, as a drug for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample; or Use of the crystalline form of any one of claims 11-22 or the pharmaceutical composition of claim 24 in the preparation of a medicament for inhibiting TEAD1, 2, 3 and / or 4 in a subject or sample; Preferably, TEAD1 and 3 are selectively inhibited compared to TEAD2 and 4.

28. Compounds represented by formula 6 or 7, Formula 6, Formula 7.

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