Complex ring compound, production method thereof, and use

Abstract

Disclosed herein are compounds, crystalline forms, and pharmaceutical compositions of Compound 1 and / or Compound 2. Also disclosed are methods of treating a disease or disorder (e.g., cancer or an infectious disease) comprising administering to a subject in need thereof one or more compounds or compositions of the disclosure.

Description

Technical Field

[0001] In various embodiments, the present disclosure generally relates to novel heterocyclic compounds, compositions thereof, methods for their preparation, and methods for their use, for example, inhibiting RAS and / or treating multiple types of diseases or disorders, such as pancreatic cancer, colorectal cancer, and lung cancer.

Background Art

[0002] Background RAS protein regulation is a pathway that transmits signals received from cell membrane receptors to important cells of downstream molecules (e.g., Raf, MEK, ERK, and PI3K) important for cell growth and survival. RAS circulates between an inactive state bound to GDP and an active state bound to GTP. The RAS protein has three gene subtypes, KRAS, NRAS, and HRAS, and also has high homology (>90%) in the N-terminal domain (amino acids 1-165). RAS is a frequently mutated cancer, among which KRAS accounts for 80% of all RAS mutations. KRAS mutations occur in about 60% of pancreatic cancers, 40% of colorectal cancers, 30% of lung cancers, and 20% of endometrial cancers (F. McCormick, 2017, Clin Cancer Res 21:1797-1801 (Non-Patent Document 1)). RAS hotspot mutations occur at codons 12, 13, and 61, and 75% of KRAS mutations occur at codon 12 (glycine) (D. K. Simanshu, D. V. Nissley, and F. McCormick, 2017, Cell, 170:17-33 (Non-Patent Document 2)).

[0003] There is a medical need for therapeutic treatment of cancer patients with RAS mutations (e.g., KRAS G12C mutations).

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

[0005] Brief Summary All the contents of the international application PCT / CN2019 / 087772 filed on May 21, 2019 and the international application PCT / CN2019 / 095947 filed on July 15, 2019 are incorporated herein by reference, both of which describe, as RAS inhibitors, for example, Compounds 1 and 2 as KRAS G12C inhibitors, and are described to be usable for treating various diseases or conditions, such as cancers associated with KRAS G12C mutations. TIFF0007710449000001.tif61128

[0006] In various embodiments, the present disclosure relates to Compound 1 or Compound 2, which may be, for example, in isolated form, substantially pure form and / or solid form. Further, pharmaceutical compositions containing Compound 1 or Compound 2, and methods for their manufacture and methods for their use are provided.

[0007] Some embodiments of the present disclosure relate to Compound 1, which may be, for example, in substantially pure form and / or solid form. In some embodiments, Compound 1 may be in amorphous form. In some embodiments, Compound 1 may be in crystalline form, for example, Form I, Form II, Form III or Form IV described herein. In some embodiments, Compound 1 may be substantially pure.

[0008] Some embodiments of the present disclosure relate to Compound 2, which may be, for example, in a substantially pure form and / or in a solid form. In some embodiments, Compound 2 may be in an amorphous form. In some embodiments, Compound 2 may be in a crystalline form, such as Form A, Form B, Form C, or Form D described herein. In some embodiments, Compound 2 may be substantially pure.

[0009] The compounds of the present disclosure may be used in the manufacture of pharmaceutical compositions. In some embodiments, the pharmaceutical composition may comprise one or more compounds of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof).

[0010] The pharmaceutical compositions described herein may be prepared for any suitable route of administration. In some embodiments, the pharmaceutical composition may be prepared for oral administration. For example, in some embodiments, the pharmaceutical composition may be a tablet or a capsule.

[0011] Some embodiments of the present disclosure relate to methods of using the compounds or compositions of the present disclosure. For example, in some embodiments, the present disclosure provides a method of inhibiting the KRAS G12C mutant protein in a cell, the method comprising contacting the cell with Compound 1 or 2. In some embodiments, the present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof), or a pharmaceutical composition described herein. In some embodiments, the cancer is a hematological malignancy, lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, endometrial cancer, gallbladder cancer, thyroid cancer, cholangiocarcinoma, and / or rectal cancer.

[0012] The compounds of the present disclosure may be used as monotherapy or combination therapy. For example, in some embodiments, the methods herein are used to treat cancer in a subject in need thereof, and the methods include administering to the subject an effective amount (e.g., a therapeutically effective amount) of a compound of the present disclosure. In some embodiments, the methods may further include treating the subject with other anti-cancer therapies. In some embodiments, the other anti-cancer therapies are chemotherapeutic agents, therapeutic antibodies, radiation therapy, cell therapy, or immunotherapy.

[0013] [Invention 1001] The following formula; TIFF0007710449000002.tif58128 A compound having [Invention 1002] The compound of Invention 1001 in solid form, such as amorphous form, crystalline form, or a combination thereof. [Invention 1003] Crystalline Form I, wherein (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 2, 4, 6, 8, or 10) of the following peaks: 6.9, 11.4, 11.5, 13.0, 14.8, 17.4, 17.8, 18.1, 19.4, and 22.8° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 6 or more, 8 or more, 12 or more, or all) of the following peaks: 6.9, 11.4, 11.5, 13.0, 13.9, 14.8, 16.8, 17.1, 17.4, 17.8, 18.1, 19.4, 21.0, 22.8, 23.7, 24.7, and 26.2° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 1A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 1B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of Invention 1001, characterized by [Invention 1004] Crystalline Form II, wherein (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, or 3) of the following peaks: 8.1, 15.6, and 18.8° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 6 or more, 8 or more, or all) of the following peaks: 6.2, 8.1, 9.4, 10.2, 15.6, 18.8, 19.1, 20.1, 21.0, 25.0, and 26.0° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 2A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 2B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of Invention 1001, characterized by [Invention 1005] Crystalline Form III, wherein (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, 5, or 6) of the following peaks: 6.6, 8.3, 11.5, 19.1, 19.9, and 22.0° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, or all) of the following peaks: 6.6, 8.3, 11.5, 12.1, 16.7, 19.1, 19.9, 22.0, 25.7, and 26.8° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 3A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 3B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of the present invention 1001, characterized by the above. [The present invention 1006] In crystalline form IV, (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, or 5) of the following peaks: 6.7, 8.6, 16.6, 18.9, and 19.2° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, 12 or more, or all) of the following peaks: 6.3, 6.7, 8.6, 14.6, 14.8, 15.7, 16.6, 17.6, 18.9, 19.2, 20.8, 21.2, and 23.2° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 4A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 4B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of the present invention 1001, characterized by the above. [The present invention 1007] The compound of the present invention 1001 in amorphous form. [The present invention 1008] The compound of any one of the present inventions 1001 to 1007, which is substantially pure. [The present invention 1009] A pharmaceutical composition comprising the compound of any one of the present inventions 1001 to 1008 and any pharmaceutically acceptable excipient. [The present invention 1010] The pharmaceutical composition of the present invention 1009, which comprises the compound of the present invention 1006 and does not contain or substantially does not contain the compound in a solid form other than form IV. [The present invention 1011] The following formula: TIFF0007710449000003.tif61128 A compound having the following. [The present invention 1012] The compound of the present invention 1011 in solid form, such as amorphous form, crystalline form, or a combination thereof. [The present invention 1013] In crystalline form A, (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, or 4) of the following peaks: 6.2, 12.6, 14.8, and 19.9° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 6 or more, or all) of the following peaks: 6.2, 12.6, 13.8, 14.8, 15.1, 18.0, 19.6, and 19.9° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 5A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 5B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of the present invention 1011, characterized by the above. [The present invention 1014] In crystalline form B, (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, or 5) of the following peaks: 6.2, 12.6, 14.8, 19.0, and 19.8° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 6 or more, 8 or more, or all) of the following peaks: 6.2, 12.6, 13.6, 14.5, 14.8, 17.8, 19.0, 19.8, 21.4, 26.3, 31.9, and 38.6° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 6A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 6B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) The compound of the present invention 1011, characterized by the above. [The present invention 1015] In crystalline form C, (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, or 3) of the following peaks: 6.2, 12.5, and 19.9° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, or all) of the following peaks: 6.2, 6.8, 7.3, 12.5, 14.2, 14.7, 15.7, 16.3, 19.9, 21.2, 22.9, and 26.1° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 7A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 7B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) A compound of the present invention 1011, characterized by the above [The present invention 1016] In crystalline form D, (1) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, or 4) of the following peaks: 5.6, 11.2, 16.9, and 22.6° 2θ ± 0.2°; (2) An X-ray powder diffraction (XRPD) pattern having one or more (e.g., 2 or more, 4 or more, 6 or more, or all) of the following peaks: 5.6, 11.2, 15.8, 16.1, 16.9, 21.4, 22.6, and 34.3° 2θ ± 0.2°; (3) An XRPD pattern substantially identical to that shown in Figure 8A; (4) A differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 8B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)) A compound of the present invention 1011, characterized by the above [The present invention 1017] The compound of the present invention 1011 in amorphous form. [The present invention 1018] A compound of any of the present inventions 1011 to 1017 that is substantially pure. [The present invention 1019] A pharmaceutical composition comprising a compound of any of the present inventions 1011 to 1018 and any pharmaceutically acceptable excipient. [The present invention 1020] The pharmaceutical composition of the present invention 1019, which comprises the compound of the present invention 1014 and does not contain or substantially does not contain the compound in solid forms other than form B. [The present invention 1021] A method for inhibiting the KRAS G12C mutant protein in cells, comprising A method comprising contacting the cell with a compound of any one of the present inventions 1001-1008 and 1011-1018. [Present Invention 1022] A method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound of any one of the present inventions 1001-1008 and 1011-1018 or a pharmaceutical composition of any one of the present inventions 1009-1010 and 1019-1020. [Present Invention 1023] The method of Present Invention 1022, wherein the cancer is a hematological malignancy, lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, endometrial cancer, gallbladder cancer, thyroid cancer, cholangiocarcinoma, and / or rectal cancer. [Present Invention 1024] The method of Present Invention 1022 or 1023, further comprising treating the subject by an additional therapy. [Present Invention 1025] The method of Present Invention 1024, wherein the additional therapy is a chemotherapeutic agent, a therapeutic antibody, radiation, cell therapy, or immunotherapy. [Present Invention 1026] The method of any one of Present Inventions 1023-1025, wherein the subject has a G12C mutation in KRAS, HRAS, and / or NRAS. It should be understood that the above summary and the following detailed description are both for the purpose of exemplifying and explaining the present invention and are not limiting.

Brief Description of the Drawings

[0014]

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DETAILED DESCRIPTION OF THE INVENTION

[0015] Detailed Description In various embodiments, the present disclosure relates to RAS inhibitors, such as KRAS inhibitors, and more specifically, Compounds 1 and 2, which may be, for example, in isolated form, substantially pure form and / or solid form. Compounds 1 and 2 have a pKa of 3 and generally exist in the free base form. Unless otherwise specified, it should be understood that Compounds 1 or 2 referred to herein are not salts with acids or bases and exist in their free base form. As described by way of example in the Examples section, various polymorphic forms of Compounds 1 and 2 were discovered. Among these polymorphs, Form IV of Compound 1 and Form B of Compound 2 are stable and have been found to be more suitable for various pharmaceutical uses compared to other forms of their respective compounds. Compounds 1 and 2 have the following structural formulas, respectively. TIFF0007710449000004.tif61128

[0016] As described in International Application PCT / CN2019 / 087772 filed on May 21, 2019 and PCT / CN2019 / 095947 filed on July 15, 2019 (the contents of each are incorporated herein by reference), Compounds 1 and 2 are RAS inhibitors, such as KRAS G12C inhibitors, and can be used to treat various diseases or medical conditions, such as cancers associated with the KRAS G12C mutation. Accordingly, some embodiments of the present disclosure further relate to pharmaceutical compositions comprising Compound 1 and / or Compound 2 described herein. In some embodiments, further provided is a method of treating or preventing a disease or medical condition associated with RAS (e.g., KRAS G12C mutation), comprising administering to a subject in need thereof a therapeutically effective amount of Compound 1 described herein or a pharmaceutical composition described herein. In some embodiments, further provided is a method of treating or preventing a disease or medical condition associated with RAS (e.g., KRAS G12C mutation), comprising administering to a subject in need thereof a therapeutically effective amount of Compound 2 described herein or a pharmaceutical composition described herein.

[0017] Compound 1 In some embodiments, the present disclosure relates to Compound 1. Compound 1 and its synthesis are described in International Application PCT / CN2019 / 087772 filed on May 21, 2019 and / or PCT / CN2019 / 095947 filed on July 15, 2019, and all the contents of each are incorporated herein by reference in their entirety. Compound 1 should be understood in its free base form, distinct from salts with external acids or bases. Unless otherwise apparent from the context, as described herein, Compound 1 should be understood in its free base form.

[0018] In some embodiments, Compound 1 may be in solid form, for example, in amorphous form, crystalline form, or a combination thereof. In some embodiments, Compound 1 may be in amorphous form. In some embodiments, Compound 1 may be in crystalline form (e.g., any one or more of crystalline forms I, II, III, and IV described herein). As used herein, when a compound (e.g., Compound 1) is described as being present as or being a particular solid form (e.g., crystalline form), in some embodiments, it should be understood that the compound is predominantly present as the particular form. However, in some embodiments, the compound can also be present as the particular form in a mixture having one or more other solid forms (including amorphous form). For example, when Compound 1 is described as being present as or being Form IV, Compound 1 is predominantly present as Form IV, e.g., more than 80 wt%, more than 90 wt%, or more than 95 wt% of Compound 1 is in Form IV, or, for example, it may contain no other solid forms recognizable by XRPD; or, in some embodiments, Compound 1 can be present as Form IV in a mixture having one or more solid forms (e.g., amorphous form).

[0019] Compound 1 as referred to herein is generally in a substantially pure form. For example, in some embodiments, by weight, by HPLC area, or both, Compound 1 can have a purity of greater than 70%, preferably greater than 90% (e.g., greater than 95%, greater than 97%, greater than 98%, greater than 98.5%). In some embodiments, Compound 1 is characterized in that the purity by weight and / or by HPLC area is about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 99%, or any range between the specified values. For example, in some embodiments, Compound 1 can be characterized in that the purity by HPLC area is about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 99%, or any range between the specified values. The substantially pure Compound 1 can be in solid form (e.g., the crystalline form, amorphous form, or a combination thereof described herein) or in solution, suspension, or another form. In some embodiments, the substantially pure Compound 1 can be in crystalline form IV. To avoid doubt, a composition containing the substantially pure Compound 1 and one or more other components herein should be understood as a mixture of the substantially pure Compound 1 herein and one or more other components, for example, the substantially pure Compound 1 can be mixed with one or more other components (e.g., solvents, pharmaceutically acceptable excipients, etc.) directly or indirectly to obtain the composition.

[0020] In some embodiments, Compound 1 is in crystalline form. In some embodiments, Compound 1 is in crystalline Form I. The characteristics of Form I include any of the characteristics described herein. In some embodiments, crystalline Form I has an X-ray powder diffraction (XRPD) pattern with one or more (e.g., 2, 4, 6, 8, or 10) of the following peaks: 6.9, 11.4, 11.5, 13.0, 14.8, 17.4, 17.8, 18.1, 19.4, and 22.8° 2θ ± 0.2°; has an X-ray powder diffraction (XRPD) pattern with one or more (e.g., 6 or more, 8 or more, 12 or more, or all) of the following peaks: 6.9, 11.4, 11.5, 13.0, 13.9, 14.8, 16.8, 17.1, 17.4, 17.8, 18.1, 19.4, 21.0, 22.8, 23.7, 24.7, and 26.2° 2θ ± 0.2°; has an XRPD pattern substantially identical to that shown in Figure 1A; has a differential scanning calorimetry (DSC) pattern substantially identical to that shown in Figure 1B; or is characterized by any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form I can be characterized by an XRPD pattern having main peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) as shown in Figure 1A or Table 1 (° 2θ ± 0.2°). More clearly, when the XRPD pattern of Form I is described as having the main peaks of Figure 1A or Table 1, or being substantially identical to Figure 1A, in some cases, the XRPD pattern does not necessarily have the same relative intensity as the corresponding peaks shown in Figure 1A or Table 1. In some cases, regardless of their relative intensities, the XRPD pattern may include peaks at each diffraction angle (° 2θ ± 0.2°) corresponding to the peaks as shown in Figure 1A or Table 1. Similar expressions for other crystalline forms herein should be understood in the same way.In some embodiments, crystalline Form I can be characterized by an XRPD pattern having all of the following peaks: 6.9, 11.4, 11.5, 13.0, 14.8, 17.4, 17.8, 18.1, 19.4 and 22.8° 2θ ± 0.2°. In some embodiments, crystalline Form I can also be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 238.7 °C and / or a peak top temperature of about 240.9 °C. As shown in the Examples section, crystalline Form I is identified as the anhydrate. In some embodiments, crystalline Form I is substantially the same as Form I obtained in Example 3 of this application.

[0021] Compound 1 in crystalline Form I can be prepared by the methods described herein. For example, in some embodiments, 1) suspending amorphous Compound 1 in a solvent such as, for example, water, isopropanol, MTBE, THF / heptane, and EA / heptane, preferably isopropanol, to form a suspension; and 2) stirring the suspension at room temperature (RT) or with heating (e.g., 50 °C) for a defined period such as, for example, 1 day, 3 days, etc., to form Compound 1 in crystalline Form I. In some embodiments, Compound 1 in crystalline Form I can be prepared by a method comprising 1) dissolving Compound 1 in a first solvent such as, for example, ethyl acetate (EA) or tetrahydrofuran (THF) to form a solution; and then 2) adding an antisolvent such as, for example, MTBE (methyl-t-butyl ether) to the solution to precipitate Compound 1. Example 3 of this application shows an exemplary process for preparing Compound 1 in Form I.

[0022] In some embodiments, Compound 1 is in crystalline Form II. The characteristics of Form II include any of the characteristics described herein. In some embodiments, crystalline Form II has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, or 3) of the following peaks: 8.1, 15.6, and 18.8° 2θ ± 0.2°; an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 6 or more, 8 or more, or all) of the following peaks: 6.2, 8.1, 9.4, 10.2, 15.6, 18.8, 19.1, 20.1, 21.0, 25.0, and 26.0° 2θ ± 0.2°; an XRPD pattern substantially the same as that shown in Figure 2A; a differential scanning calorimetry (DSC) pattern substantially the same as the pattern shown in Figure 2B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form II can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) as shown in Figure 2A or Table 2 (° 2θ ± 0.2°). In some embodiments, crystalline Form II can be characterized by an XRPD pattern having all of the following peaks: 8.1, 15.6, and 18.8° 2θ ± 0.2°. In some embodiments, crystalline Form II can also be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 171.2°C and / or a peak top temperature of about 176.8°C; and an endothermic peak with an onset temperature of about 111.2°C and / or a peak top temperature of about 148.8°C. As shown in the Examples section, Form II is identified as a monohydrate. In some embodiments, crystalline Form II is substantially the same as the crystalline Form II obtained in Example 3 of this application.

[0023] Compound 1 in crystalline form II can be produced by the methods described herein. For example, in some embodiments, Compound 1 in crystalline form II can be produced by a method comprising: 1) dissolving Compound 1 in a first solvent (e.g., acetone) to form a solution, for example, at room temperature; and then 2) adding an antisolvent, such as water, to the solution to precipitate Compound 1. In some embodiments, the method further comprises, for example, stirring a mixture of Compound 1 in the first solvent and the antisolvent at room temperature for a period of time (e.g., 1 to 5 days) to form a suspension; and optionally filtering and drying the precipitated Compound 1. Example 3 of this application shows an exemplary process for producing Compound 1 in form II.

[0024] In some embodiments, Compound 1 is in crystalline Form III. The characteristics of Form III include any of the characteristics described herein. In some embodiments, crystalline Form III has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, 5, or 6) of the following peaks: 6.6, 8.3, 11.5, 19.1, 19.9, and 22.0° 2θ ± 0.2°; an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, or all) of the following peaks: 6.6, 8.3, 11.5, 12.1, 16.7, 19.1, 19.9, 22.0, 25.7, and 26.8° 2θ ± 0.2°; an XRPD pattern substantially the same as that shown in Figure 3A; a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 3B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form III can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (° 2θ ± 0.2°) as shown in Figure 3A or Table 3. In some embodiments, crystalline Form III can be characterized by an XRPD pattern having all of the following peaks: 6.6, 8.3, 11.5, 19.1, 19.9, and 22.0° 2θ ± 0.2°. In some embodiments, crystalline Form III can further be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 95.1 °C and / or a peak top temperature of about 114.2 °C. As shown in the Examples section, Form III is identified as a dihydrate. In some embodiments, crystalline Form III is substantially the same as the crystalline Form III obtained in Example 3 of this application.

[0025] Compound 1 in crystalline form III can be prepared by the methods described herein. For example, in some embodiments, 1) Compound 1 is dissolved in a first solvent (e.g., methanol) to form a solution, e.g., at room temperature; and then 2) an anti-solvent, e.g., water, is added to the solution to precipitate Compound 1. In some embodiments, the method further includes, e.g., stirring a mixture of Compound 1 in the first solvent and the anti-solvent at room temperature for a period of time (e.g., from 1 hour to 5 days) to form a suspension; and optionally filtering and drying the precipitated Compound 1. Example 3 of the present application shows exemplary steps for preparing Compound 1 in form III.

[0026] In some embodiments, Compound 1 is in crystalline Form IV. The characteristics of Form IV include any of the characteristics described herein. In some embodiments, crystalline Form IV has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, or 5) of the following peaks: 6.7, 8.6, 16.6, 18.9, and 19.2° 2θ ± 0.2°; an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, 12 or more, or all) of the following peaks: 6.3, 6.7, 8.6, 14.6, 14.8, 15.7, 16.6, 17.6, 18.9, 19.2, 20.8, 21.2, and 23.2° 2θ ± 0.2°; an XRPD pattern substantially the same as that shown in Figure 4A; a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 4B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form IV can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (° 2θ ± 0.2°) as shown in Figure 4A or Table 4. In some embodiments, crystalline Form IV can be characterized by an XRPD pattern having all of the following peaks: 6.7, 8.6, 16.6, 18.9, and 19.2° 2θ ± 0.2°. In some embodiments, crystalline Form IV can also be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 273.0 °C and / or a peak top temperature of about 276.0 °C. As shown in the Examples section, Form IV is identified as the anhydrate. In some embodiments, crystalline Form IV is substantially the same as the crystalline Form IV obtained in Example 3 of this application.

[0027] Compound 1 in crystalline form IV can be produced by the methods described herein. For example, in some embodiments, Compound 1 in crystalline form IV can be prepared by a method comprising: 1) dissolving Compound 1 in a first solvent (e.g., methanol) to form a solution, for example, at 50 °C; and then 2) adding an anti-solvent, e.g., heptane, to the solution to precipitate Compound 1. In some embodiments, the method further comprises stirring the mixture of Compound 1 in the first solvent and the anti-solvent for a period of time (e.g., from 0.2 hours to 2 hours); and optionally filtering and drying the precipitated Compound 1. In some embodiments, Compound 1 in crystalline form IV can be prepared by a method comprising: 1) dissolving Compound 1 in a suitable solvent, e.g., ethyl acetate, under heating conditions (e.g., up to the boiling point of the solvent) to form a solution; and then 2) cooling the solution, e.g., to room temperature or below, e.g., to 0 - 10 °C, to precipitate Compound 1. Example 3 of the present application shows an exemplary process for producing Compound 1 in form IV.

[0028] As will be described in detail in the Examples section, compared to various other forms, Compound 1 in form IV can be more suitable for various pharmaceutical applications. For example, based on interconversion studies, it was shown that all forms I, II, and III convert to form IV at 50 °C or room temperature in an anhydrous solvent system, indicating that form IV is the stable form. When evaluating the form stability of form IV in an aqueous solution, it was shown that form IV is stable at room temperature or 50 °C for 3 days and does not form a hydrate.

[0029] In some embodiments, Compound 1 may be in an amorphous form. The amorphous form of Compound 1 can be produced by various methods described herein. As shown in the present text, compared to various other forms, amorphous Compound 1 can be more suitable for some pharmaceutical applications, because amorphous Compound 1 has better solubility in bio-related liquids compared to the tested crystalline forms.

[0030] The method for producing various crystal forms of Compound 1 according to the present invention generally uses one or more solvents. Suitable solvents are generally known and include, but are not limited to, THF, toluene, MeOH, ethanol, n-propanol, isopropanol, isobutanol, methyl-t-butyl ether, ethyl ether, isoamyl alcohol, butyl acetate, ethyl formate, 1,4-dioxane, n-butanol, t-butanol, n-heptane, cyclohexane, methyl isobutyl ketone, xylene, isobutyl acetate, 2-butanone, acetonitrile, acetone, ethyl acetate, isopropyl acetate, and water. The solvents may be used alone or in combination of multiple types. Crystal technologies are well-known in the art. For example, Compound 1 may be slurried in one or more solvents at room temperature or with heating; Compound 1 may be heated in one or more solvents and then cooled; Compound 1 may be dissolved in a solvent and then an anti-solvent may be added; other technologies, such as solid / liquid diffusion or liquid / liquid diffusion, may also be used. The starting Compound 1 may be, but is not limited to, an amorphous solid. In some embodiments, the starting Compound 1 may also be in a crystal form, such as Form I. In some embodiments, the starting Compound 1 may also be a combination of an amorphous solid and a crystal form.

[0031] In some embodiments, the present disclosure further provides a solid form of Compound 1 that can be produced by any applicable method described in the Examples section.

[0032] Compound 2 In some embodiments, the present disclosure relates to Compound 2. Compound 2 and its synthesis are described in International Application PCT / CN2019 / 087772 filed on May 21, 2019 and / or PCT / CN2019 / 095947 filed on July 15, 2019, and all the contents of each are incorporated herein by reference. Compound 2 should be understood in its free base form, distinct from salts with external acids or bases. Unless otherwise apparent from the context, as described herein, Compound 2 should be understood in its free base form.

[0033] In some embodiments, Compound 2 may be in a solid form, for example, an amorphous form, a crystalline form, or a combination thereof. In some embodiments, Compound 1 may be in an amorphous form. In some embodiments, Compound 2 may be in a crystalline form (e.g., any one or more of the crystalline forms A, B, C, and D described herein). As used herein, when Compound 2 is described as being present as or being a particular solid form (e.g., a crystalline form), in some embodiments, it should be understood that Compound 2 is predominantly present as the particular form. However, in some embodiments, Compound 2 can also be present as the particular form in a mixture having one or more other solid forms (including the amorphous form). For example, when Compound 2 is described as being present as or being Form B, Compound 2 is predominantly present as Form B, for example, more than 80% by weight, more than 90% by weight, or more than 95% by weight of Compound 2 is in Form B, or, for example, it may contain no other solid form recognizable by XRPD; or, in some embodiments, Compound 2 can be present as Form B in a mixture having one or more solid forms (e.g., an amorphous form).

[0034] Compound 2 as referred to herein is generally in a substantially pure form. For example, in some embodiments, by weight, by HPLC area or both, Compound 2 can have a purity of more than 70%, preferably more than 90% (e.g., more than 95%, more than 97%, more than 98%, more than 98.5%). In some embodiments, Compound 2 can be characterized in that its purity by weight and / or by HPLC area is about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 99%, or any range between the specified values. For example, in some embodiments, Compound 2 can be characterized in that its purity by HPLC area is about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 99% or any range between the specified values. The substantially pure Compound 2 can be in solid form (e.g., the crystalline form, amorphous form or a combination thereof described herein) or in solution, suspension or another form. In some embodiments, the substantially pure Compound 2 can be in crystalline form B. To avoid doubt, the composition containing the substantially pure Compound 2 and one or more other components herein should be understood as a mixture of the substantially pure Compound 2 herein and one or more other components herein. For example, the substantially pure Compound 2 can be mixed with one or more other components (e.g., solvents, pharmaceutically acceptable excipients, etc.) directly or indirectly to obtain the composition.

[0035] In some embodiments, Compound 2 is in a crystalline form. In some embodiments, Compound 2 is in crystalline Form A. The characteristics of Form A include any of the characteristics described herein. In some embodiments, crystalline Form A has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, or 4) of the following peaks: 6.2, 12.6, 14.8, and 19.9° 2θ ± 0.2°; (2) an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 6 or more, or all) of the following peaks: 6.2, 12.6, 13.8, 14.8, 15.1, 18.0, 19.6, and 19.9° 2θ ± 0.2°; (3) an XRPD pattern substantially the same as that shown in Figure 5A; (4) a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 5B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form A can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (° 2θ ± 0.2°) as shown in Figure 5A or Table 9. In some embodiments, crystalline Form A can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.6, 14.8, and 19.9° 2θ ± 0.2°. In some embodiments, crystalline Form A can be characterized by an XRPD pattern not having the following peaks: 7.3, 14.2, 15.7, and 16.3° 2θ ± 0.2°. In some embodiments, crystalline Form A can be characterized by an XRPD pattern not having two or more, three or more, or all of the following peaks: 7.3, 14.2, 15.7, and 16.3° 2θ ± 0.2°. In some embodiments, crystalline Form A can be characterized by an XRPD pattern not having the following peaks: 14.5, 15.6, 20.2, and 38.6° 2θ ± 0.2°.In some embodiments, crystalline Form A can be characterized by an XRPD pattern that does not have two or more, three or more, or all of the following peaks: 14.5, 15.6, 20.2 and 38.6° 2θ ± 0.2°. In some embodiments, crystalline Form A can further be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 286.4 °C and / or a peak top temperature of about 289.2 °C. In some embodiments, crystalline Form A is substantially the same as crystalline Form A obtained in Example 5 or 6 of the present application.

[0036] Compound 2 in crystalline Form A can be produced by the methods described herein. For example, in some embodiments, a method comprising slurrying Compound 2 in a solvent such as ethyl acetate (EA or EtOAc); or 1) dissolving Compound 2 in a suitable solvent (e.g., acetone, THF, 2-butanone and / or dichloromethane (DCM)), and then 2) adding an anti-solvent (e.g., n-heptane) to precipitate Compound 2 can be used to produce Compound 2 in crystalline Form A. Examples 5 and 6 of the present application show exemplary steps for producing Compound 2 in Form A.

[0037] In some embodiments, Compound 2 is in crystalline Form B. The characteristics of Form B include any of the characteristics described herein. In some embodiments, crystalline Form B has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, 3, 4, or 5) of the following peaks: 6.2, 12.6, 14.8, 19.0, and 19.8° 2θ ± 0.2°; has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 6 or more, 8 or more, or all) of the following peaks: 6.2, 12.6, 13.6, 14.5, 14.8, 17.8, 19.0, 19.8, 21.4, 26.3, 31.9, and 38.6° 2θ ± 0.2°; has an XRPD pattern substantially the same as that shown in Figure 6A; has a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 6B; or can be characterized by any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form B can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (2θ ± 0.2°) as shown in Figure 6A or Table 10. In some embodiments, crystalline Form B can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.6, 14.8, 19.0, and 19.8° 2θ ± 0.2°. In some embodiments, crystalline Form B can be characterized by an XRPD pattern having at least one (e.g., 1, 2, 3, 4, 5, or 6) of the following peaks: 14.5, 17.8, 21.4, 26.3, 31.9, and 38.6° 2θ ± 0.2°. In some embodiments, crystalline Form B can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.6, 14.8, 19.0, and 19.8° 2θ ± 0.2° and having at least one (e.g., 1, 2, 3, 4, 5, or 6) of the following peaks: 14.5, 17.8, 21.4, 26.3, 31.9, and 38.6° 2θ ± 0.2°.In some embodiments, crystalline form B has all of the following peaks: 6.2, 12.6, 14.8, 19.0 and 19.8° 2θ ± 0.2°, has two or more, three or more, or all of the following peaks: 14.5, 15.6, 20.2 and 38.6° 2θ ± 0.2°; and can be characterized by an XRPD pattern that does not have both or either of the following peaks: 7.3 and 14.2° 2θ ± 0.2°. In some embodiments, crystalline form B can further be characterized by a DSC pattern having an endothermic peak at an onset temperature of about 289.0 °C and / or a peak top temperature of about 290.1 °C. In some embodiments, crystalline form B is substantially identical to crystalline form B obtained in Example 6 of this application.

[0038] Compound 2 in crystalline form B can be produced by the methods described herein. For example, in some embodiments, compound 2 in crystalline form B is prepared by: 1) dissolving compound 2 in a first solvent (e.g., methanol) to form a solution, e.g., at room temperature; and then 2) adding an anti-solvent, such as water, to the solution to precipitate compound 2. In some embodiments, the method further includes, for example, stirring a mixture of compound 2 in the first solvent and the anti-solvent at room temperature for a certain period (e.g., 1 to 24 hours) to form a suspension; and optionally filtering and drying the precipitated compound 2. In some embodiments, compound 2 in crystalline form B can be produced in various crystalline forms. For example, in some embodiments, compound 2 in crystalline form B is prepared by: 1) suspending compound 2 (e.g., form A) in a solvent (e.g., methanol) to form a suspension; and 2) stirring the suspension at room temperature (RT) or with heating, e.g., at 50 °C, for a certain period, such as 1 day, 3 days, etc., to form compound 2 in crystalline form B. The concentration range of the suspension with the solvent may be 15 to 100 mg / ml, e.g., about 100 mg / mL. Example 6 of this application shows an exemplary process for producing compound 2 in form B.

[0039] In some embodiments, Compound 2 is in crystalline Form C. The characteristics of Form C include any of the characteristics described herein. In some embodiments, crystalline Form C has an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 1, 2, or 3) of the following peaks: 6.2, 12.5, and 19.9° 2θ ± 0.2°; an X-ray powder diffraction (XRPD) pattern having one or more (e.g., 4 or more, 8 or more, or all) of the following peaks: 6.2, 6.8, 7.3, 12.5, 14.2, 14.7, 15.7, 16.3, 19.9, 21.2, 22.9, and 26.1° 2θ ± 0.2°; an XRPD pattern substantially the same as that shown in Figure 7A; a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 7B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form C can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (° 2θ ± 0.2°) as shown in Figure 7A or Table 11. In some embodiments, crystalline Form C can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.5, and 19.9° 2θ ± 0.2°. In some embodiments, crystalline Form C can be characterized by an XRPD pattern having at least one (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or all) of the following peaks: 7.3, 14.2, 14.7, 15.7, 16.3, 19.9, 21.2, 22.9, and 26.1° 2θ ± 0.2°. In some embodiments, crystalline Form C can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.5, and 19.9° 2θ ± 0.2° and having at least one (e.g., 1, 2, 3, or 4) of the following peaks: 7.3, 14.2, 20.8, and 26.1° 2θ ± 0.2°.In some embodiments, crystalline Form C can be characterized by an XRPD pattern having all of the following peaks: 6.2, 12.5 and 19.9° 2θ ± 0.2°; and having one or both of the following peaks: 7.3 and 14.2° 2θ ± 0.2°. In some embodiments, crystalline Form C can further be characterized by a DSC pattern having an endothermic peak at an onset temperature of about 288.7 °C and / or a peak top temperature of about 289.4 °C. In some embodiments, crystalline Form C is substantially identical to crystalline Form C obtained in Example 6 of the present application.

[0040] Compound 2 in crystalline Form C can be produced by the methods described herein. Example 6 of the present application shows exemplary steps for producing Compound 2 in Form C.

[0041] In some embodiments, Compound 2 is in crystalline Form D. The characteristics of Form D include any of the characteristics described herein. In some embodiments, crystalline Form D has an X-ray powder diffraction (XRPD) pattern with one or more (e.g., 1, 2, 3, or 4) of the following peaks: 5.6, 11.2, 16.9, and 22.6° 2θ ± 0.2°; an X-ray powder diffraction (XRPD) pattern with one or more (e.g., 2 or more, 4 or more, 6 or more, or all) of the following peaks: 5.6, 11.2, 15.8, 16.1, 16.9, 21.4, 22.6, and 34.3° 2θ ± 0.2°; an XRPD pattern substantially the same as that shown in Figure 8A; a differential scanning calorimetry (DSC) pattern substantially the same as that shown in Figure 8B; or any combination thereof (e.g., (1) and (4), (2) and (4), (1) and (2) and (4), or (3) and (4)). In some embodiments, crystalline Form D can be characterized by an XRPD pattern having major peaks (e.g., peaks with a relative intensity of 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more) (° 2θ ± 0.2°) as shown in Figure 8A or Table 12. In some embodiments, crystalline Form D can be characterized by an XRPD pattern having all of the following peaks: 5.6, 11.2, 16.9, and 22.6° 2θ ± 0.2°. In some embodiments, crystalline Form D can be characterized by an XRPD pattern having at least one (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the following peaks: 5.6, 11.2, 15.8, 16.1, 16.9, 21.4, 22.6, and 34.3° 2θ ± 0.2°. In some embodiments, crystalline Form D can also be characterized by a DSC pattern having an endothermic peak with an onset temperature of about 286.9°C and / or a peak top temperature of about 289.0°C, and an endothermic peak with an onset temperature of about 133.7°C and / or a peak top temperature of about 140.8°C. In some embodiments, crystalline Form D is substantially the same as the crystalline Form D obtained in Example 6 of this application.

[0042] Compound 2 in crystalline form D can be prepared by the methods described herein. For example, in some embodiments, 1) dissolving Compound 2 in a first solvent such as isopropanol or isobutanol to form a first solution, for example, a saturated solution; then, 2) dissolving Compound 2 in a second solvent such as 2-butanone, acetone or THF to form a second solution, for example, a saturated solution; 3) mixing the first solution and the second solution; and 4) precipitating Compound 2 by slowly evaporating the solvent. In some embodiments, the first and second solvents may be isopropanol and 2-butanone, isopropanol and THF, isopropanol and acetone, or isobutanol and THF. Example 6 of the present application shows an exemplary process for preparing Compound 2 in form D.

[0043] As will be described in detail in the Examples section, compared to various other forms, Compound 2 in form B can be more suitable for various pharmaceutical applications. As can be seen from the results of solid stability, form B is stable both physically and chemically for 7 days under the conditions of 40 °C / 75% RH (relative humidity), and this crystalline form does not change at 92.5% RH for 10 days and at 60 °C for 7 days. Also, based on the interconversion studies, form B is more stable than forms A and C.

[0044] In some embodiments, Compound 2 may be in an amorphous form. The amorphous form of Compound 2 can be prepared by various methods described herein.

[0045] In the methods for producing various crystalline forms of Compound 2 in this specification, generally, one or more solvents are used. Suitable solvents are generally known and include, but are not limited to, THF, toluene, MeOH, ethanol, n-propanol, isopropanol, isobutanol, methyl-t-butyl ether, ethyl ether, isoamyl alcohol, butyl acetate, ethyl formate, 1,4-dioxane, n-butanol, t-butanol, n-heptane, cyclohexane, methyl isobutyl ketone, xylene, isobutyl acetate, 2-butanone, acetonitrile, acetone, ethyl acetate, isopropyl acetate, and water. The solvents may be used alone or in combinations of multiple types. Crystallization techniques are well-known in the art. For example, Compound 2 may be slurried in one or more solvents at room temperature or with heating; Compound 2 may be heated in one or more solvents and then cooled; Compound 2 may be dissolved in a solvent and then an anti-solvent may be added; other techniques, such as solid / liquid diffusion or liquid / liquid diffusion, may also be used. The starting Compound 2 may be, but is not limited to, an amorphous solid or may be in a crystalline form, such as Form A. In some embodiments, the starting Compound 2 may be a combination of an amorphous solid and a crystalline form.

[0046] In some embodiments, the present disclosure further provides a solid form of Compound 2 that can be produced by any of the application methods described in the Examples section.

[0047] In some embodiments, the present disclosure further relates to any product produced by any of the methods described herein and methods of using these products.

[0048] Pharmaceutical composition In various embodiments, the present disclosure further provides a pharmaceutical composition comprising a compound of the present disclosure, such as Compound 1 (e.g., Form IV) or Compound 2 (e.g., Form B) described herein, and optionally a pharmaceutically acceptable excipient. Non-limiting suitable excipients include, for example, absorption enhancers, antioxidants, binders, buffers, carriers, coating agents, colorants, diluents, disintegrants, emulsifiers, extenders, fillers, flavoring agents, humectants, lubricants, fragrances, preservatives, propellants, release agents, sterilizing agents, sweetening agents, solubilizing agents, wetting agents, and mixtures thereof, such as encapsulating agents or additives. Reference can also be made to Remington′s The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2005; incorporated herein by reference), which discloses various excipients for preparing pharmaceutical compositions and known techniques for their manufacture.

[0049] In some embodiments, the present disclosure provides a pharmaceutical composition comprising one or more compounds of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof). Generally, the pharmaceutical composition comprises a therapeutically effective amount of one or more compounds of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof) and any pharmaceutically acceptable excipient or carrier. In some embodiments, the pharmaceutical composition comprises one or more substantially pure compounds (e.g., Compound 1 and / or 2) described herein. In some embodiments, the pharmaceutical composition comprises one or more solid forms selected from the group consisting of Compound 1 in Form I, Compound 1 in Form II, Compound 1 in Form III, Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form A, Compound 2 in Form B, Compound 2 in Form C, Compound 2 in Form D, and amorphous Compound 2.

[0050] In some specific embodiments, the pharmaceutical composition comprises Form IV of Compound 1. In some specific embodiments, the active ingredient in the pharmaceutical composition may comprise, consist essentially of, or consist of Form IV of Compound 1. In some embodiments, Compound 1 is present in the pharmaceutical composition essentially as Form IV. For example, at least 80% (e.g., at least 85%, at least 90%, at least 95% of the total weight of Compound 1) of Compound 1 is present in the pharmaceutical composition as Form IV. In some embodiments, the pharmaceutical composition substantially does not contain any other solid form (e.g., other crystalline forms) of Compound 1. In some embodiments, the pharmaceutical composition does not contain or substantially does not contain crystalline forms of Compound 1 other than Form IV. For example, in some embodiments, the pharmaceutical composition may contain less than 10%, less than 5%, less than 2%, less than 1% or an undetectable amount of crystalline forms of Compound 1 other than Form IV, based on the total weight of Compound 1.

[0051] In some specific embodiments, the active ingredient in the pharmaceutical composition may comprise, consist essentially of, or consist of Compound 1 in Form IV, amorphous form or a mixture thereof. In some embodiments, Compound 1 may be present in the pharmaceutical composition as a mixture of Form IV and amorphous form of Compound 1. For example, at least 80% (e.g., at least 85%, at least 90%, at least 95% by the total weight of Compound 1) of Compound 1 may be present in the pharmaceutical composition as Form IV or amorphous form.

[0052] In some specific embodiments, the pharmaceutical composition comprises Form B of Compound 2. In some specific embodiments, the active ingredient in the pharmaceutical composition may comprise, consist essentially of, or consist of Form B of Compound 2. In some embodiments, Compound 2 is present in the pharmaceutical composition essentially as Form B, for example, at least 80% (e.g., at least 85%, at least 90%, at least 95% based on the total weight of Compound 2) of Compound 2 is present in the pharmaceutical composition as Form B. In some embodiments, the pharmaceutical composition substantially does not contain any other solid form of Compound 2, for example, other salts or other crystalline forms. In some embodiments, the pharmaceutical composition does not contain or substantially does not contain crystalline forms of Compound 2 other than Form B, for example, in some embodiments, the pharmaceutical composition may contain less than 10%, less than 5%, less than 2%, less than 1% or an undetectable amount of crystalline forms of Compound 2 other than Form B based on the total weight of Compound 2.

[0053] In some specific embodiments, the active ingredient in the pharmaceutical composition may comprise, consist essentially of, or consist of Compound 2 in Form B, amorphous form or a mixture thereof. In some embodiments, Compound 2 may be present in the pharmaceutical composition as a mixture of Form B and amorphous form of Compound 2, for example, at least 80% (e.g., at least 85%, at least 90%, at least 95% by the total weight of Compound 2) of Compound 2 may be present in the pharmaceutical composition as Form B or amorphous form.

[0054] Generally, a pharmaceutical composition provides a compound of the present disclosure in an effective amount. In some embodiments, the effective amount is a therapeutically effective amount (e.g., an amount effective to treat cancer comprising a G12C mutation of KRAS, HRAS, and / or NRAS (e.g., a KRAS G12C mutation) in a subject in need thereof). As used herein, a therapeutically effective amount of a compound of the invention is an amount effective to treat a disease or condition described herein, and may depend on the subject being treated, the disease or condition being treated and its severity, the composition containing the compound, the time of administration, the route of administration, the duration of treatment, the efficacy of the compound (e.g., the efficacy to inhibit KRAS G12C), its clearance rate, and whether another medicament is taken concomitantly.

[0055] The pharmaceutical compositions described herein can be manufactured by any method known in the pharmacological arts. Generally, such manufacturing methods include mixing an active ingredient, such as a salt of the invention, with a carrier or excipient and / or one or more other accessory ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into the required single-dose or multi-dose units.

[0056] The pharmaceutical compositions may be manufactured, packaged, and / or sold as a single unit dose and / or as a plurality of single unit doses. A "unit dose" is a pharmaceutical composition containing an isolated amount of a predetermined quantity of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to the subject and / or a convenient fraction of the dose, such as half or one-third of the dose.

[0057] The relative amounts of the active ingredient, pharmaceutically acceptable excipient, and / or any other ingredient in the pharmaceutical compositions described herein can vary depending on the identity, size, and / or condition of the subject to be treated, and further depending on the route by which the composition is to be administered. The composition may contain from 0.1% to 100% (w / w) of the active ingredient.

[0058] Pharmaceutically acceptable excipients that can be used in the manufacture of the pharmaceutical composition herein include, for example, inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. The composition may contain excipients such as, for example, cocoa butter, suppository wax, colorants, coating agents, sweeteners, flavoring agents, and fragrances.

[0059] The pharmaceutical composition can be prepared so as to be used for any route of administration such as oral administration. Generally, the pharmaceutical composition is in a solid dosage form. However, in some embodiments, other dosage forms such as liquids, suspensions, semi-solid dosage forms can also be used.

[0060] Solid dosage forms for oral administration include, for example, capsules, tablets, pills, powders, and granules. Among such solid dosage forms, there are an active ingredient and at least one inert pharmaceutically acceptable excipient or carrier (e.g., sodium citrate, dicalcium phosphate) and / or (a) fillers or extenders (e.g., starch, lactose, sucrose, glucose, mannitol, silicic acid), (b) binders (e.g., carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, acacia), (c) humectants (e.g., glycerin), (d) disintegrants (e.g., agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates, and sodium carbonate), (e) dissolution inhibitors (e.g., paraffin), (f) absorption promoters (e.g., quaternary ammonium compounds), (g) wetting agents (e.g., cetyl alcohol, glyceryl monostearate), (h) absorbents (e.g., kaolin, bentonite), (i) lubricants (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof) are mixed. In the case of capsules, tablets, and pills, the dosage form may contain a buffer.

[0061] Solid compositions of the same type can be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycol. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be manufactured with coatings and outer shells (e.g., enteric coatings and other coatings well known in the pharmaceutical field). These may optionally contain light protectants and may be compositions that release the active ingredient only in a part of the intestinal tract or, preferably, optionally with a delay. Examples of encapsulating compositions that can be used include, for example, polymeric substances and waxes. Solid compositions of the same type can be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar, high molecular weight polyethylene glycol.

[0062] The active ingredient (e.g., the compounds of the present disclosure) may be in a microencapsulated form having one or more of the above excipients. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be manufactured with coatings and outer shells (e.g., enteric coatings and other coatings well known in the pharmaceutical field). In such solid dosage forms, the active ingredient can be mixed with at least one inert diluent (e.g., sucrose, lactose, starch). Such dosage forms can generally contain substances other than inert diluents, such as tableting lubricants and other tableting aids such as magnesium stearate, microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage form may contain a buffering agent. These may optionally contain light protectants and may be compositions that release the active ingredient only in a part of the intestinal tract or, preferably, optionally with a delay. Examples of encapsulating compositions that can be used include, for example, polymeric substances and waxes.

[0063] The description of the pharmaceutical compositions provided herein is mainly for pharmaceutical compositions suitable for administration to humans, although these compositions are generally suitable for administration to various animals. It is well known to modify pharmaceutical compositions suitable for administration to humans to those suitable for administration to various animals, and a veterinarian with general skills can design and / or perform this modification by general experiments. For veterinary use, based generally on veterinary practice, the compounds of the present disclosure can be administered as a formulation that is suitably accepted. A veterinarian can readily determine the optimal dosing plan and route of administration for a particular animal.

[0064] The compounds of the present disclosure are generally formulated in dosage units so as to be easy to administer and to have a uniform dosage. However, it should be understood that the total daily usage of the compositions described herein will be determined by a physician within the scope of reasonable medical judgment. For a particular subject or organism, the specific effective dosage level of treatment depends on various factors including the severity of the disease and condition being treated, the activity of the specific active ingredient being used, the specific composition being used, the age, weight, general health, sex and diet of the subject, the time of administration, the route of administration and the excretion rate of the specific active ingredient being used, the duration of the treatment, the medicaments used in combination with or concurrently with the specific active ingredient being used, and similar factors well known in the medical arts.

[0065] In some embodiments, all necessary components for treating KRAS-related diseases using the compounds of the present invention alone or in combination with another agent or intervention that has been conventionally used for treating such diseases may be packaged in a kit. Specifically, in some embodiments, the present invention provides a kit for treating an intervention of a disease, comprising a buffered solution and other components for producing a deliverable form of the medicament, and / or a device for delivering the medicament; and / or any agent used for combination therapy with the compounds of the present disclosure, and / or a treatment instruction for the disease packaged together with the medicament. The instructions may be fixed to any tangible medium such as printed paper, may be a computer-readable magnetic or optical medium, or may direct to refer to a data source of a remote computer such as a web page accessible via the Internet.

[0066] Treatment method The compounds of the present disclosure and the pharmaceutical compositions described herein may be used to treat and / or prevent diseases or conditions associated with RAS (e.g., KRAS G12C).

[0067] In some embodiments, the present disclosure provides a method for inhibiting cell signaling via RAS, comprising contacting a cell with an effective amount of one or more compounds of the present disclosure (e.g., Compound 1 and / or 2). Inhibition of signaling via RAS can be evaluated and demonstrated by multiple types of methods known in the art. Non-limiting examples are: (a) a decrease in the GTPase activity of RAS; (b) a decrease in the GTP-binding affinity or an improvement in the GDP-binding affinity; (c) an improvement in the K of GTP or the K of GDP off of an improvement or GDP offDecrease; (d) Decrease in the level of a signaling molecule downstream of the RAS pathway, for example, decrease in the level of pMEK, pERK or pAKT; and / or (e) Decrease in the binding of the RAS complex to downstream signaling molecules (including but not limited to Raf). Kits and commercially available assays may be used for one or more of the above determinations.

[0068] In some embodiments, the present disclosure provides a method for inhibiting KRAS, HRAS, and / or NRAS G12C in a cell, the method comprising contacting the cell with an effective amount of one or more compounds of the present disclosure (e.g., Compound 1 and / or 2).

[0069] In some embodiments, the present disclosure provides a method for treating a disease or disorder in a subject in need thereof, such as cancer associated with a G12C mutation in KRAS, HRAS, and / or NRAS, for example, cancer associated with KRAS G12C. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof) or a therapeutically effective amount of a pharmaceutical composition described herein.

[0070] In some embodiments, provided is a method of treating cancer, the method comprising administering to a subject in need thereof an effective amount of any of the compounds of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof) or a pharmaceutical composition comprising a compound of the present disclosure. In some embodiments, the cancer comprises a G12C mutation of KRAS, HRAS, and / or NRAS, e.g., a KRAS G12C mutation. Determining whether a tumor or cancer comprises a G12C mutation of KRAS, HRAS, and / or NRAS is known in the art and is described, for example, in US2018 / 0334454. In various embodiments, the cancer may be pancreatic cancer, endometrial cancer, colorectal cancer, or lung cancer (e.g., non-small cell lung cancer). In some embodiments, the cancer is a hematological cancer (e.g., as described herein). In some embodiments, the cancer is a polyposis associated with MYH. In some embodiments, the cancer is gallbladder cancer, thyroid cancer, or cholangiocarcinoma.Non-limiting examples of cancer further include acute myeloid leukemia, adolescent cancer, pediatric adrenocortical carcinoma, AIDS-related cancers (such as lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical teratoid tumor, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, heart tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ductal carcinoma in situ (DCIS), embryonal tumor, central nervous system cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrosarcoma of bone, gallbladder cancer, gastric cancer, gastrointestinal cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic cell, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell carcinoma and occult primary central line cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell tumor, mucormycosis, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of bone and osteosarcoma, nasal and paranasal cavity cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, gastric cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, renal pelvis and ureteral transitional cell carcinoma, trophoblastic tumor, pediatric atypical cancer, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer or virus-induced cancer.

[0071] In some embodiments, the present disclosure provides a method of treating a disease or disorder (e.g., cancer as described herein) in a subject in need thereof, the method comprising determining whether the subject has a G12C mutation of KRAS, HRAS, and / or NRAS (e.g., a KRAS G12C mutation), and, if it is determined that the subject has a G12C mutation of KRAS, HRAS, and / or NRAS (e.g., a KRAS G12C mutation), administering to the subject a therapeutically effective amount of at least one compound of the present disclosure (e.g., Compound 1 in Form IV, amorphous Compound 1, Compound 2 in Form B, amorphous Compound 2, or any combination thereof) or a pharmaceutical composition comprising at least one compound of the present disclosure.

[0072] G12C mutations of KRAS, HRAS, and / or NRAS have also been identified in hematologic malignancies (e.g., cancers affecting the blood, bone marrow, and / or lymph nodes). Thus, some embodiments relate to a method of treating a hematologic malignancy in a subject in need thereof, generally comprising administering to the subject a compound of the present disclosure (e.g., as a pharmaceutical composition). Such malignancies include, but are not limited to, leukemias and lymphomas, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myeloid leukemia (CML), acute monocytic leukemia (AMoL), and / or other leukemias. In some embodiments, the hematologic malignancy can further include lymphomas (e.g., Hodgkin lymphoma or non-Hodgkin lymphoma), plasma cell malignancies (e.g., multiple myeloma, mantle cell lymphoma, and Waldenström macroglobulinemia).

[0073] The compounds of the present disclosure can be used as monotherapy or combination therapy. In some embodiments, the combination therapy includes treating a subject with a chemotherapeutic agent, a therapeutic antibody, radiation therapy, cell therapy, or immunotherapy. In some embodiments, the compounds of the present disclosure can also be co-administered, simultaneously or sequentially in any order, with other pharmaceutically active compounds to a subject in need thereof (e.g., a subject suffering from cancer associated with the KRAS G12C mutation described herein). In some embodiments, the other pharmaceutically active compounds can be chemotherapeutic agents, therapeutic antibodies, etc. Any known chemotherapeutic agent can be used in combination with the compounds of the present disclosure. In some embodiments, the compounds of the present disclosure can also be used in combination with radiation therapy, hormone therapy, cell therapy, surgery, and immunotherapy well-known to those skilled in the art.

[0074] As used herein, administration is not limited to any particular route of administration. For example, in some embodiments, administration can be oral, nasal, transdermal, pulmonary, inhalational, buccal, sublingual, intraperitoneal, subcutaneous, intramuscular, intravenous, rectal, intrapleural, intrathecal, or parenteral. In some embodiments, oral administration is performed.

[0075] The dosing regimen, including the dose, can vary and be adjusted according to the subject being treated, the disease or condition being treated and its severity, the composition containing the compound, the time of administration, the route of administration, the duration of treatment, the efficacy of the compound, the clearance rate, and whether another medicament is co-administered simultaneously.

[0076] Definitions As used herein, "(one or more) compounds of the present disclosure" means Compound 1, Compound 2, its isolated form, its substantially pure form, its solid form (including crystalline form, amorphous form, hydrate, and / or solvate).

[0077] As used herein, the term "about," when used to limit amounts related to the present invention, means, for example, normal testing and processing; unintentional errors during such testing and processing; differences in the manufacture, origin, or purity of the components used in the present invention, etc., which can result in variations in the number of numerical values that can occur. As used herein, an "about" specific value further includes the said specific value. For example, "about 10%" includes 10%. Whether or not limited by the term "about," the claims include equivalent forms of the recited numbers. In one embodiment, the term "about" refers to within 20% of the reported numerical value.

[0078] As used herein, the term "treat" (including "treat," "treating," "treatment," etc.) means to eliminate, alleviate, or relieve a disease or medical condition and / or the symptoms associated therewith. Although not excluded, the treatment of a disease or medical condition does not necessarily require complete elimination of the said disease, medical condition, or the symptoms associated therewith. As used herein, the term "treat" (including "treat," "treating," "treatment," etc.) may include "preventive treatment," which means reducing the risk of, or the likelihood of recurrence or progression of, a disease or medical condition in a subject who is not yet affected but is at risk, or has had a previous recurrence or progression of the disease or medical condition. The synonyms of the term "treat" are understood to mean administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure.

[0079] As used herein, the term "therapeutically effective amount" means an amount of a therapeutic agent (e.g., any one or more of the compounds of the present disclosure) sufficient to relieve one or more symptoms of a disease or medical condition (e.g., cancer associated with the KRAS G12C mutation), prevent the appearance or progression of a disease or medical condition, or cause regression or cure of a disease or medical condition.

[0080] As used herein, the term "subject" (herein also sometimes referred to as "patient") means an animal that is the subject of treatment, observation, or experiment, preferably a mammal, and most preferably a human. In any of the embodiments described herein, the subject may be a human.

Example

[0081] Example 1. General Method Materials: Starting materials, reagents, solvents, etc. are generally commercially available. 1 1H NMR was performed on a Bruker Advance 300 equipped with an autosampler (B-ACS 120).

[0082] Powder X-ray diffraction (XRPD): Solid samples were examined with an X-ray diffractometer (Bruker D8 advance). The system was equipped with a LynxEye detector. The wavelength of the X-ray was 1.5418 Å. The samples were scanned from 3 to 40° 2θ with a step size of 0.02° 2θ. The tube voltage and current were 40 KV and 40 mA, respectively.

[0083] Polarizing microscopy analysis (PLM): Optical microscopy was performed with a polarizing microscope ECLIPSE LV100POL (Nikon, JPN).

[0084] TGA analysis: TGA was performed on a TGA Q500 or Discovery TGA 55 (TA Instruments, US). The sample was placed in an open aluminum weighing dish, automatically weighed, and then inserted into the TGA furnace. The sample was heated from room temperature (RT) to the final temperature at a rate of 10 °C / min.

[0085] DSC analysis: DSC was performed on a DSC Q200 or Discovery DSC 250 (TA Instruments, US). The sample was placed in an aluminum pinhole-sealed pan and the weight was accurately recorded. The sample was heated from 25 °C to the final temperature at a rate of 10 °C / min.

[0086] Dynamic moisture sorption analysis (DVS): Method 1. It was used for the study of the solid of Compound 1. Moisture adsorption / desorption data were collected with a DVS Intrinsic (SMS, UK). The sample was placed in a gravimetric sample chamber and weighed automatically. The sample was dried at 40 °C until dm / dt was less than 0.002%, and then cooled to 25 °C. The instrument parameters were set as follows. Step time (minutes): 60 min Sample temperature: 25 °C Cycle: Full cycle Adsorption: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 Desorption: 80, 70, 60, 50, 40, 30, 20, 10, 0 Save data rate: 5 s Total flow rate: 200 sccm Total flow rate after experiment: 200 sccm

[0087] Method 2. It was used for the study of the solid of Compound 2. Moisture adsorption / desorption data were collected with an IGAsorp dynamic moisture sorption analyzer. The sample was placed in a gravimetric sample chamber and weighed automatically. The sample was dried at 50 °C until the humidity was less than 0.3%, and then cooled to 25 °C. The instrument parameters were set as follows. Sample temperature: 25 °C Temperature stability: 0.1 °C / min Flow rate: 250 mL / min Scan: 2 Mode: F1 Minimum time: 30 min Timeout: 120 min End: 98% Start: Adsorption scan Adsorption: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 Desorption: 80, 70, 60, 50, 40, 30, 20, 10, 0

[0088] HPLC analysis: Method 1. The following is a representative HPLC method, which can be used, for example, to analyze the purity, solubility, and stability of Compound 1 as referred to herein. TIFF0007710449000005.tif63170

[0089] Method 2. HPLC analysis was performed using an Agilent HPLC 1260 series instrument. The representative HPLC method can be used to analyze the purity, solubility, and stability studies of Compound 2. TIFF0007710449000006.tif60170

[0090] Example 2. Preparation and Solid Characterization of Compound 1 TIFF0007710449000007.tif156165

[0091] Compound 1-1 was prepared according to the synthesis process of Compound 1-1 in Example 1 of WO2020233592A1.

[0092] Step 1: A mixture of 3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (10 g, 42.18 mmol) and di-t-butyl dicarbonate (12.6 g, 57.73 mmol) in toluene (10 mL) was heated at 80 °C for 5 hours. The mixture was concentrated and the residue was purified by silica gel flash column chromatography (from ethyl acetate / petroleum ether = 0 / 1 to 1 / 3) to obtain 1-a.

[0093] Step 2: At room temperature, oxalyl chloride (11 g, 88 mmol) and 15 drops of dry DMF were added to a suspension of 2,5,6-trichloronicotinic acid (10 g, 44 mmol) in dichloromethane (100 mL). After 30 minutes, the resulting solution was concentrated to obtain a residue, which was dissolved in 1,4-dioxane (40 mL). 100 mL of aqueous ammonia (28% NH3 aqueous solution) was added dropwise at 0 °C, and the reaction mixture was stirred again for 10 minutes, filtered, and washed with water. The filter cake was recovered and lyophilized to obtain 1-2.

[0094] Step 3: A solution of 1-2 (30 g, 133.07 mmol) in DCE (300 mL) was treated with oxalyl chloride (33.7 g, 265.52 mmol). The mixture was stirred at 80 °C for 45 minutes and then concentrated. The residue was dissolved in dichloromethane (60 mL) and concentrated. The residue was dissolved in THF (120 mL), cooled to -10 °C, and a solution of 1-1 (23.3 g, 139.96 mmol) in THF (120 mL) was added. The resulting mixture was stirred at room temperature for 2 hours. The mixture was extracted between EtOAc and water. The organic layer was washed with brine and concentrated. The residue was slurried in EtOAc / PE (1 / 10) and filtered. The filter cake was dried to obtain 1-3.

[0095] Step 4: K3PO4 (6.5 g, 30.62 mmol) was added to a stirred solution of 1-3 (10 g, 25.63 mmol) in DMF (60 mL). The resulting mixture was then stirred at room temperature (RT) for 2 hours. The reaction was quenched with HCl (1 N) and filtered. The filter cake was washed with water and then slurried in MeCN. It was filtered and the filter cake was dried to obtain 1-4.

[0096] Step 5: POCl 3( 7.9 g, 51.53 mmol) was added dropwise to a solution of 1-4 (10 g, 25.63 mmol) and DIEA (8.3 g, 64.22 mmol) in MeCN (50 mL) at room temperature. The resulting solution was stirred at 80 °C for 45 minutes, then at -10 °C a solution of DIEA (8.3 g, 64.22 mmol) and (2R,5S)-t-butyl-2,5-dimethylpiperazine-1-carboxylate (6.6 g, 30.80 mmol) in MeCN (50 mL) was added dropwise. After stirring at room temperature for 1 hour, the reaction was quenched with ice water and the mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel flash column chromatography (ethyl acetate / petroleum ether = 0 / 1 to 3 / 1) to obtain 1-5.

[0097] Step 6: At 80 °C, under a nitrogen atmosphere, a mixture of 1-5 (2 g, 3.41 mmol), 1-a (1.38 g, 4.09 mmol), Pd(dppf)Cl2 (250 mg, 0.34 mmol) and K3PO4 (1.45 g, 6.83 mmol) in toluene (20 mL) was stirred for 2 hours. The mixture was diluted with water and then extracted with ethyl acetate. The combined organic layers were dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel flash column chromatography (ethyl acetate / petroleum ether = 0 / 1 to 3 / 1) to obtain 1-6.

[0098] Step 7: A solution of 1-6 (5 g, 6.57 mmol) in DCM (25 mL) was treated with TFA (9 g), and the mixture was stirred at 25 °C for 3 hours. The reaction was quenched with aqueous Na2CO3 solution, and the mixture was separated. The organic layer was washed twice with water and then concentrated to obtain 1-7.

[0099] Step 8: At -10 °C, acryloyl chloride (322 mg, 3.56 mmol) was added to a mixture of 1-7 (2 g, 3.56 mmol) and DIPEA (550 mg, 4.26 mmol) in THF (40 mL). The mixture was stirred for 1 hour and then quenched with aqueous citric acid solution. The mixture was extracted with EtOAc, and the organic layer was washed with water. The organic layer was concentrated, and the residue was purified by silica gel flash column chromatography (ethyl acetate / petroleum ether = 0 / 1 to 3 / 1) to obtain 1. Compound 1 was lyophilized from MeCN / H2O to obtain an amorphous form. LCMS (ESI, m / z): [M+H] +=615.3; HNMR(400 MHz, DMSO-d6, ppm): δ 8.74(s, 1H), 8.52 - 8.35(m, 1H), 7.16 - 7.07(m, 1H), 6.94 - 6.76(m, 1H), 6.51(d, J = 6.0 Hz, 1H), 6.38(t, J = 6.6 Hz, 1H), 6.20(dd, J = 12.6, 1.8 Hz, 1H), 5.76(dd, J = 12.6, 1.6 Hz, 1H), 4.91 - 4.80(m, 2H), 4.51 - 3.50(m, 6H), 1.91 - 1.65(m, 1H), 1.45 - 1.15(m, 6H), 1.10 - 0.70(m, 8H). FNMR(376 MHz, DMSO-d6, ppm): δ -114.30(1F).

[0100] It was found that the compound 1 thus obtained was an amorphous solid. By TGA, two weight loss stages of 1.4% before 200 °C and 0.6% thereafter were shown, and by DSC, one broad endothermic peak was observed at 35 °C. Also, the Tg of such an amorphous solid observed by DSC was about 154 °C. TIFF0007710449000008.tif33139

[0101] Example 3. Polymorph Screening of Compound 1 In this example, the polymorphs of compound 1 were screened.

[0102] Evaporative Crystallization: An appropriate amount of compound 1 was added to 2.5 mL of 13 solvents (MeOH, EtOH, IPA (isopropanol), IBA (isobutanol), MEK (methyl ethyl ketone or 2-butanone), THF (tetrahydrofuran), ACN (acetonitrile), MTBE (methyl-t-butyl ether), acetone, water, toluene, EA (ethyl acetate), IPAC (isopropyl acetate)) to prepare suspensions respectively. After stirring, 2 mL of 13 drug suspensions were each filtered. Then, the filtrates were used for binary solvent screening or single solvent evaporation studies.

[0103] Binary evaporation study: The saturated drug solution (filtered solution) was dispersed in a 96-well plate. Two different filtered solutions were placed in each well, and the volume of each filtered solution was 100 μL. The plate was covered with a sealing film with pinholes and evaporated in a laboratory ventilation case during operation under environmental conditions. All the obtained samples were in a glassy state or amorphous, and no crystalline samples were produced.

[0104] Single-solvent evaporation study: The saturated drug solution (filtered solution) was used for slow evaporation study. After drying, the solid samples were examined by XRPD. Only two crystalline samples were obtained. Sample 1 was obtained in IPA, and Form II was obtained in isobutanol, and the other samples were all in a glassy state or amorphous.

[0105] Slurrying study: Suspensions prepared with IPA, water, MTBE, and heptane were slurried at room temperature for 3 days and then at 50 °C for 1 day. Solid samples were collected by filtration and analyzed by XRPD at specific times. When new XRPD patterns were identified, the samples were further analyzed by DSC and TGA.

[0106] Alternatively, amorphous Compound 1 (about 20 mg) was weighed into a vial, and then 0.5 mL of the selected mixed solvent was added. The suspension was stirred at room temperature for several days or at 50 °C for 1 day. Solid samples were collected by filtration and analyzed by XRPD at specific times. When new XRPD patterns were identified, the samples were further analyzed by DSC and TGA.

[0107] Anti-solvent precipitation: Amorphous Compound 1 (about 20 mg) was dissolved in 0.1 or 0.2 mL of a solvent at room temperature, and then the anti-solvent was slowly added until precipitation appeared or up to 1 mL of the anti-solvent. When precipitation occurred, corresponding identifications were made for the product. Amorphous Compound 1 has high solubility in most solvents, while having low solubility in MTBE, water, and heptane.

[0108] In polymorph screening, five XRPD patterns (Figure 11) were generated and four forms were identified, including two anhydrates and two hydrates. Sample 1 was the XRPD pattern of the wet cake and was unstable. Samples 2 to 5 were identified as Forms I, II, III, and IV, respectively. The details are as follows.

[0109] Sample 1 was the wet cake of a sample prepared by evaporation or slurrying in IPA. Pattern 1 could be converted to Form I during air drying, and pure Sample 1 was not produced. Therefore, Sample 1 was an unstable solvate / hydrate and was not designated as a form.

[0110] Form I was obtained by slurrying in water, IPA, MTBE, THF / heptane, and EA / heptane, or by antisolvent precipitation from EA / MTBE and THF / MTBE. Only Form I produced in IPA has a single sharp melting peak, which is thought to be due to residual solvent or crystal quality.

[0111] Scale-up test of Form I: At room temperature, an amorphous compound (about 250 mg) was slurried in 1.5 mL of IPA for 3 days. The sample was collected by filtration, dried at 50 °C for 5 hours, and then identified. Form I (about 180 mg) was successfully produced with a yield of 72%. The thermal curve showed that there was a single melting peak, the onset temperature was 239 °C, and the weight loss from room temperature to 200 °C was 0.17% (Figure 1B). NMR showed that there was no residual solvent and Form I was an anhydrous form.

[0112] Representative XRPD and DSC patterns of Form I are shown in Figures 1A - 1B. The list of XRPD peaks is shown in Table 1 below.

[0113] (Table 1) Table of XRPD peak values for Form I. TIFF0007710449000009.tif108166

[0114] Form II was produced in a mixed solvent of acetone - heptane or acetone - water, and acetone - water was selected as the solvent to produce Form II.

[0115] Scale - up test of Form II: At room temperature, amorphous Compound 1 (about 100 mg) was dissolved in 0.2 mL of acetone. After adding 0.2 mL of water, a sticky sample appeared immediately. However, after stirring at room temperature for 3 days, it became a suspension. The sample was collected by filtration and dried at 50 °C for 5 hours and then identified. Form II (about 65 mg) was successfully produced with a yield of 63%.

[0116] From the DSC results, it was found that there were two endothermic peaks with onset temperatures of 111 °C and 171 °C respectively. By TGA, a weight loss of about 3.1% before 150 °C (Figure 2B) was observed, but 0.1% of residual acetone was detected by NMR. Therefore, the weight loss was due to water, and Form II was a monohydrate (the theoretical water content of the monohydrate is 2.8%). However, it was not a stable hydrate, and dehydration occurred at low temperature.

[0117] Representative XRPD and DSC patterns of Form II are shown in Figures 2A - 2B. The list of XRPD peaks is shown in Table 2 below.

[0118] (Table 2) Table of XRPD peak values of Form II. TIFF0007710449000010.tif97166

[0119] Form III was obtained by precipitation from MeOH / water. According to the following procedure, Form III (115 mg) was successfully produced with a yield of 73%.

[0120] Scale-up test of Form III: At room temperature, amorphous Compound 1 (about 100 mg) was dissolved in 0.3 mL of MeOH, and then 0.2 mL of water was slowly added. Immediately, a sticky sample appeared, and after stirring for 2 hours, a solid appeared. After stirring for another 2 hours, the solid sample was collected by filtration and then dried overnight at room temperature.

[0121] From the thermal curve, it was found that there was one endothermic peak with a starting temperature of 95 °C and a 5% weight loss before 150 °C (Figure 3B). No residual solvent was detected by NMR. Therefore, Form III is a dihydrate (the theoretical water content of the dihydrate is 5.5%), still unstable, and dehydration occurred at low temperature, converting to the amorphous form after dehydration.

[0122] Representative XRPD and DSC patterns of Form III are shown in Figures 3A - 3B. The list of XRPD peaks is shown in Table 3 below.

[0123] (Table 3) Table of XRPD peak values of Form III. TIFF0007710449000011.tif129166

[0124] Form IV was obtained by cooling crystallization of Form I in EA as the starting material, or by anti-solvent precipitation of amorphous Compound 1 in acetone / heptane at 50 °C as the starting material. Finally, Form IV was produced in acetone - heptane.

[0125] Scale-up test of Form IV: At 50 °C, amorphous Compound 1 (about 100 mg) was dissolved in 0.2 mL of acetone, and then 0.4 mL of heptane was added. After stirring at 50 °C for 0.5 hours, a solid appeared. After stirring for 1 hour, the sample was collected by filtration and then identified after vacuum drying at 50 °C for 3 hours. Form IV (about 80 mg) was successfully produced with a yield of 80%.

[0126] By DSC and TGA, one melting peak at an onset temperature of 273 °C and a 0.33% weight loss before 200 °C were observed respectively (Figure 4B). By NMR, about 0.22% residual acetone was detected. Form IV should be the anhydrous form but contained residual solvent. The DVS results (Figure 4C) showed that Form IV was slightly hygroscopic, absorbed about 1.37% water at 80% RH, and the crystal form did not change after the DVS test.

[0127] Representative XRPD and DSC patterns of Form IV are shown in Figures 4A - 4B. The list of XRPD peaks is shown in Table 4 below.

[0128] (Table 4) Table of XRPD peak values of Form IV. TIFF0007710449000012.tif118163

[0129] First, as summarized in Table 5, four forms were obtained and identified. Forms I and IV were anhydrates. Two hydrates, Form II (monohydrate) and Form III (dihydrate), were both unstable and dehydration occurred at low temperature.

[0130] (Table 5) Characterization results of each form TIFF0007710449000013.tif83170

[0131] Example 4. Interconversion and solubility studies of solid forms of Compound 1 Interconversion study: Equal amounts (about 7 mg) of Forms I, II, and III were mixed together and then suspended in 0.5 mL of various solvents at room temperature or 50 °C respectively. The residual solid was collected by filtration and identified at appropriate times. Also, Form IV (about 12 mg) was slurried in 0.5 mL of water or a MeOH / water (1 / 4) solution at room temperature or 50 °C for 3 days. The residual solid was collected by filtration and analyzed by XRPD.

[0132] From the results, it was found that, except for IPA and MeOH / water (1 / 4), a mixture of Forms I, II and III converted to Form IV at room temperature (Table 6). In IPA, the wet cake was still Pattern 1, highly likely to be an IPA solvate, and Form II was obtained in a water-containing solvent system at room temperature. Therefore, Form IV was a stable form in a solvent system at high temperature and an anhydrous solvent system at room temperature.

[0133] (Table 6) Interconversion study TIFF0007710449000014.tif32135

[0134] Form IV was slurried in a solvent system containing water to confirm whether it was a stable form in water. As can be seen from the results, Form IV was slurried in water or methanol / water at room temperature and 50 °C for 3 days without change, and no hydrate was formed. Therefore, Form IV was stable for 3 days in a water-containing system without hydrate crystal seeds.

[0135] Solubility study: The solubility of Form IV was measured in three biorelevant media (simulated gastric fluid (SGF), fasted-state simulated intestinal fluid (FaSSIF) and fed-state simulated intestinal fluid (FeSSIF)) and water. The solubility of amorphous Compound 1 was measured in FaSSIF. Form IV (about 10 mg) was weighed and placed in 2 mL of the four media, and suspensions were prepared respectively. Also, about 10 mg of amorphous raw material was added to 2 mL of FaSSIF. Then, all suspensions were shaken at 37 °C at a speed of 200 rpm for 24 hours. At 0.5, 2 and 24 hours, about 0.7 mL of each suspension was filtered. The filtrate was analyzed by HPLC and pH meter, and the remaining solid was examined by XRPD at 24 hours.

[0136] As shown in Table 7, the solubility of Form IV in FaSSIF was lower than that of the amorphous raw material, and the solubility decreased from 0.3 mg / mL to 0.02 mg / mL. In other media, Form IV also had a relatively low solubility, <0.07 mg / mL.

[0137] (Table 7) Solubility Results TIFF0007710449000015.tif53170

[0138] Stability study: An appropriate amount of Form IV was left standing for 14 days under the conditions of 40 °C / 75% RH, 60 °C and 25 ± 2 °C / 92.5% RH. Then, the samples were measured by HPLC and XRPD to confirm chemical and physical stability. The results are shown in Table 8.

[0139] (Table 8) Results of Stability Evaluation TIFF0007710449000016.tif19161

[0140] After storing Form IV at 60 °C for 2 weeks, new impurities were detected and the impurities increased with time. On the other hand, the purity of the raw material Form IV was only about 98.3%. Regarding the physical stability test, after 14 days under the test conditions, XRPD did not change, and it was found that Form IV had physical stability under all three test conditions during the test period.

[0141] Furthermore, the mechanical stability of Form IV was measured. In such a case, an appropriate amount of Form IV was ground for 2 and 5 minutes, and then measured by XRPD to examine its crystal form. As can be seen from the results, no new form was produced and the crystallinity decreased during grinding.

[0142] Example 5. Preparation and Solid Characterization of Compound 2 TIFF0007710449000017.tif119165

[0143] Step 1: To a mixture of 2,6-dichloro-5-fluoronicotinic acid (23 g, 0.11 mol) in dichloromethane (300 mL) was added dimethylformamide (0.2 mL). Oxalyl chloride (33 g, 0.26 mol) was slowly added thereto within 30 minutes at room temperature. The mixture was stirred at room temperature for 1 hour and then concentrated to obtain an oily substance, which was dissolved in 1,4-dioxane (50 mL). Ammonia water (150 mL) was added to the solution within 30 minutes at 0 °C. The resulting mixture was stirred at 0 °C for 30 minutes and then filtered. The filter cake was washed with cold water (50 mL) and dried to obtain 2-1.

[0144] Step 2: The solution of 2-1 (11 g, 52.6 mmol) in 1,2-dichloroethane (80 mL) was treated with oxalyl chloride (8.68 g, 68.4 mmol). The mixture was stirred at 80 °C for 45 minutes and the reaction product was concentrated. The residue was dissolved in acetonitrile (100 mL), cooled to -10 °C, and a solution of 1-1 (9.6 g, 55.2 mmol) in THF (30 mL) was added. The resulting mixture was stirred at room temperature for 2 hours. The solution was diluted with saturated aqueous NaHCO3 and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (from petroleum ether to petroleum ether / ethyl acetate = 4 / 1) to obtain 2-2.

[0145] Step 3: To a solution of 2-2 (14 g, 34.13 mmol) in DMF (84 mL) stirred at 20 - 30 °C was added K3PO4 (8.7 g, 40.99 mmol). The resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with HCl (1 N) and filtered. The solid was washed with water and then slurried in MeCN. The mixture was filtered and the filter cake was dried to obtain 2-3.

[0146] Step 4: At room temperature, POCl3 (8.2 g, 53.48 mmol) was added dropwise to a solution of 2-3 (16.6 g, 44.41 mmol) and DIEA (8.6 g, 66.54 mmol) in MeCN (83 mL). The resulting mixture was stirred at 80 °C for 45 minutes, and then at -10 °C, a solution of DIEA (8.6 g, 66.54 mmol) and (2R,5S)-tert-butyl-2,5-dimethylpiperazine-1-carboxylate (9.5 g, approximately 44.33 mmol) in MeCN (33 mL) was added dropwise. After stirring at room temperature for 1 hour, the reaction was quenched with an aqueous Na2CO3 solution, and the mixture was filtered. The solid was slurried in water and then filtered. The solid was dried to obtain 2-4.

[0147] Step 5: A mixture of 2-4 (50 g, 87.71 mmol), 3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (25 g, 105.45 mmol), Pd(dppf)Cl2 (1.3 g, 0.18 mmol), potassium acetate (KOAc) (17.2 g, 175.26 mmol), and water (1.6 g) in 1,4-dioxane (500 mL) was stirred at 80 °C for 2 hours under a nitrogen atmosphere. The mixture was cooled and then filtered. The filtrate was added to water, precipitated, and then filtered. The solid was dissolved in DMC and washed with an aqueous Na2CO3 solution. The organic phase was solvent-exchanged to toluene by distilling it multiple times. The mixture was heated to 80 °C in toluene and then cooled to 20 °C to crystallize. It was filtered, and the filter cake was dried to obtain 2-5.

[0148] Step 6: Treat the mixture of 2-5 (10 g, 15.51 mmol) in DCM (50 mL) with TFA (21.2 g) and stir the mixture at 25 °C for 3 h. Quench the reaction with aqueous Na2CO3 solution and separate the mixture. Wash the organic layer twice with water and then concentrate it. Dissolve the residue in EtOH and add an EtOH solution of fumaric acid. Filter the mixture and wash the solid with EtOH. Add the solid to aqueous Na2CO3 solution and extract the mixture with DCM. Wash the organic layer with water and then concentrate it. Exchange the solvent of the residue by distillation multiple times to toluene and then filter it. Wash the filter cake with heptane and dry it to obtain 2-6.

[0149] Step 7: Add a solution of 3-chloropropionyl chloride (2.28 g, 17.96 mmol) in THF (100 mL) to the mixture of 2-6 (10 g, 18.36 mmol) and DIPEA (2.6 g, 20.12 mmol) in THF (100 mL) at -10 °C. Stir the mixture for 1 h and then quench it with aqueous citric acid solution. Extract the mixture with EtOAc and wash the organic layer with water. Exchange the solvent of the organic layer by distillation multiple times to toluene and then filter it. Wash the filter cake with heptane and dry it to obtain 2-7.

[0150] Step 8: Stir the mixture of 2-7 (10 g, 15.75 mmol) and DBU (4.79 g, 31.26 mmol) in THF (70 mL) and DMSO (30 mL) at 20 °C for 1 h. Add EtOAc and aqueous citric acid solution. Separate the mixture and wash the organic layer with water. Exchange the solvent of the organic layer by distillation multiple times to isopropanol and then filter it. Wash the filter cake with water and dry it to obtain 2. LCMS (ESI, m / z): [M+H] += 599.1. HNMR (400 MHz, methanol-d4, ppm): δ 8.73 (s, 1H), 8.26 - 8.22 (m, 1H), 7.15 - 7.09 (m, 1H), 6.84 - 6.74 (m, 1H), 6.53 (d, J = 8.4 Hz, 1H), 6.42 - 6.38 (m, 1H), 6.30 - 6.24 (m, 1H), 5.83 - 5.78 (m, 1H), 5.01 (brs, 1H), 4.91 - 4.83 (m, 1H), 4.53 - 4.29 (m, 2H), 3.96 - 3.89 (m, 1.5H), 3.54 - 3.50 (m, 0.5H), 1.82 - 1.75 (m, 1H), 1.73 - 1.66 (m, 1H), 1.47 (d, J = 6.8 Hz, 3H), 1.37 - 1.27 (m, 3H), 1.16 - 1.05 (m, 4H), 1.03 - 0.97 (m, 2H), 0.88 - 0.83 (m, 2H). FNMR (376 MHz, methanol-d4, ppm): δ -114.9 (1F), -125.6 (1F).

[0151] The compound 2 produced by the above process was slurried in EtOAc and then filtered to obtain the compound 2 in crystalline form A. 1 By H-NMR, about 1.1% of residual EtOAc was detected, which corresponded to the weight loss at 120 - 290 °C in TGA (Figure 5B). By DSC (Figure 5B), two overlapping endothermic peaks were observed. The compound 2 in form A was heated up to 250 °C, and there was no change in the DSC curve of the residual solid. It was found that the overlapping peaks were due to the melting associated with the crystal form conversion. Therefore, the starting material was an anhydride.

[0152] Form A was very soluble in DCM (>92 mg / mL) and soluble in methanol, butanone, THF, ACN, and acetone (20 - 33 mg / mL). In other solvents, form A was substantially insoluble.

[0153] Representative XRPD and DSC patterns of form A are shown in Figures 5A - 5B. A list of XRPD peaks is shown in Table 9 below.

[0154] (Table 9) Table of XRPD peak values of Form A. TIFF0007710449000018.tif92169

[0155] Example 6. Polymorph screening of Compound 2 In this example, the polymorphs of Compound 2 were screened. Starting from Form A of Compound 2, following the same procedure as in Example 3, it was slurried in commonly used solvents and solvent mixtures, cooled, evaporated to crystallize, and polymorph screening was carried out by anti-solvent precipitation and mechanical treatment methods.

[0156] Slurrying in single solvents: 15 to 20 mg of Form A of Compound 2 was added to different solvents (MeOH, EtOH, IPA (isopropanol), IBA (isobutanol), MEK (methyl ethyl ketone or 2-butanone), THF (tetrahydrofuran), ACN (acetonitrile), MTBE (methyl-t-butyl ether), acetone, water, toluene, EA (ethyl acetate) or IPAC (isopropyl acetate)), and suspensions with a concentration of 15 to 100 mg / mL were prepared. The suspensions were stirred at 50 °C for 1 day or at room temperature for 3 days respectively. Solid samples were collected by filtration and analyzed by XRPD. When a new XRPD pattern was identified, the sample was dried under vacuum at 50 °C overnight, and then the sample was further analyzed by DSC and TGA.

[0157] Five new patterns were identified by XRPD and designated as Patterns 2, 3, 4, 5, and 6, and Pattern 2 was obtained from most of the solvents. The wet cake obtained by slurrying in EtOH at 50 °C for 1 day was identified as Pattern 3 and converted to Pattern 4 after drying. The wet cake obtained by slurrying in IPA at room temperature or 50 °C was identified as Pattern 5 and converted to Pattern 6 after drying.

[0158] Slurrying in mixed solvents (organic solvent / water) at room temperature and 50 °C: Approximately 20 mg of Compound 2 in Form A was added to 1 mL of each of the MeOH / H2O (1 / 9) and ACN / H2O (1 / 9) mixed solvents to prepare suspensions with a concentration of 20 mg / mL. The suspensions were stirred at 50 °C or room temperature respectively. The solid samples were collected by filtration and analyzed by XRPD. If a new XRPD pattern was identified, the sample was dried under vacuum at 50 °C overnight and then further analyzed by DSC and TGA. In all experiments, Pattern 1 (Form A) was obtained in every case and no new pattern was obtained.

[0159] Evaporative crystallization in binary solvents: Thirteen saturated drug solutions (filtrates) in MeOH, EtOH, IPA (isopropanol), IBA (isobutanol), MEK (methyl ethyl ketone or 2-butanone), THF (tetrahydrofuran), ACN (acetonitrile), MTBE (methyl-t-butyl ether), acetone, water, toluene, EA (ethyl acetate) or IPAC (isopropyl acetate) were dispensed into a 96-well plate. Each well contained two different filtrates, and the volume of each filtrate was 100 μL. The plate was covered with a sealing film with pinholes and evaporated in an operating laboratory ventilation case under environmental conditions. All solids in sufficient amounts were analyzed by XRPD.

[0160] Three new patterns were identified and designated as Pattern 7, Pattern 9, and Pattern 10 respectively. Pattern 7 was obtained by evaporation from EtOH and is similar to Pattern 4. Pattern 9 was obtained from 96-well plates in IPA / 2-butanone, IPA / THF, IPA / acetone, and isobutanol / THF. Pattern 10 was obtained by evaporation from acetone / iPrOAc and converted to Pattern 2 (Form B) after drying. All other samples obtained on the 96-well plate were in a glassy state.

[0161] Evaporation crystallization in a single solvent: Approximately 0.5 mL of saturated drug solutions (filtrates) of 13 types, namely MeOH, EtOH, IPA (isopropanol), IBA (isobutanol), MEK (methyl ethyl ketone or 2 - butanone), THF (tetrahydrofuran), ACN (acetonitrile), MTBE (methyl - t - butyl ether), acetone, water, toluene, EA (ethyl acetate) or IPAC (isopropyl acetate), were evaporated with the lid open under environmental conditions in an operating laboratory ventilation case. All solids in sufficient amounts were analyzed by XRPD.

[0162] Cooling crystallization: 15 - 20 mg of Compound 2 in Form A was added to a vial. Different solvents (MeOH, THF, acetone, ACN, 2 - butanone, ethyl acetate or EtOH) were added in 100 - 500 μL aliquots while stirring at 50 °C until the solution became clear. After filtration at 50 °C, the filtrate was cooled to room temperature or 5 °C. The solid samples were collected by filtration and analyzed by XRPD. Patterns 2 and 6 were discovered from the experiments conducted in MeOH and EtOH respectively.

[0163] Antisolvent precipitation: Based on the expected solubility results, antisolvent precipitation was carried out using six selected good solvents (acetone, MeOH, THF, ACN, 2 - butanone and DCM) and five antisolvents (toluene, iPrOAc, MEBE, water and n - heptane) at room temperature. Saturated drug solutions were prepared in the good solvents. Then, the antisolvent was added gradually at room temperature until turbidity or up to 15V. Pattern 1 (Form A) was produced from most of the precipitation experiments, except for MeOH / MTBE, MeOH / water and 2 - butanone / water (in which Pattern 2 (Form B) was obtained). New patterns were identified from acetone / water and THF / water. It was designated as Pattern 8 and converted to Pattern 1 (Form A) after drying.

[0164] A total of 10 XRPD patterns were discovered. All the XRPD patterns were similar, having a main peak at about 6° 2θ, but pattern 9 showed a peak shift. Interestingly, the crystalline samples showed very similar XRPD patterns, indicating that they had similar structures. Based on the identification data, three anhydrous forms including the initial form were determined and designated as form A (pattern 1), B (pattern 2) and form C (pattern 6); furthermore, a solvate was discovered and designated as form D (pattern 9). The other patterns were not designated as forms because they were impurity phases or unstable.

[0165] Identification was performed using form B (pattern 2) obtained by slurrying (about 100 mg / mL) in MeOH at 50 °C. By TGA, no weight loss was observed from 60 to 280 °C, the melting endothermic peak of DSC was at 289 - 290 °C, and the enthalpy was 95 J / g (Figure 6B). The sample was an anhydrate and was designated as form B.

[0166] Form B was the most stable form, and a small-scale scale-up test was performed on it and used for DVS analysis, solubility and stability studies. 103.02 mg of compound 2 in form A was dissolved in 5 mL of MeOH (48 V) at room temperature. After filtration, 10 mL of water was added to the filtrate. Immediately, precipitation occurred. The suspension was stirred at room temperature for about 2 hours. The solid sample was collected by filtration and vacuum dried at 50 °C overnight. 94.16 mg of solid was obtained, and the yield was 91%.

[0167] Form B was slightly hygroscopic, absorbing about 0.84% water at 0 - 80% RH (Figure 6C), and there was no change in the crystal form after the DVS test (Figure 6D).

[0168] Typical XRPD and DSC patterns of form B are shown in Figures 6A - 6B. The list of XRPD peaks is shown in Table 10 below.

[0169] (Table 10) Table of XRPD peak values of form B. TIFF0007710449000019.tif147162

[0170] From the slurrying experiment in EtOH at 50 °C, Pattern 3 was identified. After vacuum drying overnight at 50 °C, it was converted to Pattern 4 and finally to Form B (Pattern 2) after desolvation or dehydration.

[0171] Pattern 4 was plate-like crystals. By TGA, a 1.76% weight loss was observed at 110 - 240 °C, corresponding to a small broad endothermic peak due to desolvation or dehydration observed by DSC. After desolvation, it was converted to Form B, with a melting point of 288 - 290 °C and an enthalpy of 85 J / g. Pattern 4 may be a solvate or hydrate, but after comparison with Pattern 6, it was found to be a mixture of Pattern 6 and another pattern.

[0172] From the slurrying experiment in IPA at 50 °C, Pattern 5 was obtained. After vacuum drying overnight at 50 °C, it was converted to Pattern 6 and finally to Form B during the heating period.

[0173] Form C (Pattern 6): By TGA, a 0.9% weight loss was observed at 112 - 232 °C, presumed to be due to solvent loss. In DSC, an exothermic peak was observed at 206 - 221 °C, followed by a melting peak at 288.7 - 289.4 °C (Figure 7B), and it was found that there was a potential crystal transformation during the heating process. Pattern 6 may be an anhydrate and was designated as Form C (Figure 7A). The XRPD pattern of Form C was similar to that of Pattern 4, but other peaks in Pattern 4 were observed by XRPD, and it was found that Pattern 4 was a mixture of Form C and a solvate or hydrate.

[0174] Representative XRPD and DSC patterns of Form C are shown in Figures 7A - 7B. A list of XRPD peaks is shown in Table 11 below.

[0175] (Table 11) Table of XRPD peak values for Form C. TIFF0007710449000020.tif92166

[0176] Pattern 7 was obtained by evaporation from EtOH and it was similar to Pattern 4. By DSC, after heating to 250 °C, it was converted to Form B (Pattern 2). 1 By 1H-NMR, about 1.3% residual EtOH was detected, corresponding to the weight loss at 102 - 210 °C in TGA. By DSC, two endothermic peaks due to desolvation and melting were observed. Pattern 7 may be a mixed form and is similar to Pattern 4.

[0177] Pattern 8 was obtained by anti-solvent precipitation from acetone and water. After vacuum drying overnight at 50 °C, it was converted to Form A. Pattern 8 may have been a mixed form of Form A and an unstable solvate. Further analysis was not performed.

[0178] Form D (Pattern 9) was obtained by evaporation from IPA / 2-butanone, IPA / THF, IPA / acetone and isobutanol / THF in a 96-well plate. After vacuum drying overnight at 50 °C, there was no change in the crystal form but the crystallinity decreased. By DSC, after heating to 250 °C, it was converted to Form B. About 4% weight loss was observed at 107 - 223 °C in TGA (Figure 8B). By NMR, 4.8% (about 0.5 mol) of solvent was observed and the weight loss in TGA was estimated to be due to the solvent. By DSC, two endothermic peaks due to desolvation and melting were observed (Figure 8B). Pattern 9 is a solvate and was designated as Form D.

[0179] Representative XRPD and DSC patterns of Form D are shown in Figures 8A - 8B. A list of XRPD peaks is shown in Table 12 below.

[0180] (Table 12) Table of XRPD peak values of Form D. TIFF0007710449000021.tif124166

[0181] Pattern 10 was obtained by evaporation from 96-well plates with acetone / iPrOAc and acetone / water. After drying under vacuum at 50 °C overnight, it was converted to Form B. Pattern 10 is an unstable form and no further analysis was performed.

[0182] A total of 10 XPRD patterns were identified, including 3 anhydrates, 1 solvate, 2 mixed forms and 4 unstable patterns. The XRPD patterns of Form A and Form B are very similar and clearly different from Form C. The characteristic peaks of Form C were at the positions of 7.3°, 14.2°, 15.7°, 16.3°, 20.8° and 26.1° 2θ. The characteristic peaks of Form B different from Form A were mainly at the positions of 14.5°, 15.6°, 17.5°, 20.2°, 31.9° and 38.6° 2θ. Table 13 summarizes the anhydrous forms of Compound 2.

[0183] (Table 13) Summary of the anhydrous forms of Compound 2 TIFF0007710449000022.tif46128

[0184] Example 7. Interconversion and solubility studies of the solid forms of Compound 2 Interconversion studies: Competitive slurries of Form A, Form B and Form C were carried out in acetone and acetone / water (1 / 4) at room temperature or 50 °C, and the details are shown below. TIFF0007710449000023.tif42140

[0185] Form A, Form B and Form C were competitively slurried in acetone and acetone / water (1 / 4). The mixtures of different forms showed a tendency to completely convert to Form B in non-aqueous solvents after 1 day and to Form B in solvent-aqueous solutions after 7 days, with the peaks at 14.5° and 17.5° 2θ gradually increasing. The DSC of the remaining solid showed that the endothermic peak of Form A became very weak, indicating that the mixture tended to convert to Form B. From these results, Form B was a more stable form than Form A and Form C.

[0186] Solubility in biologically relevant media: Forms A and B were each added to SGF, FeSSIF, FaSSIF, and water. The suspensions were continuously shaken at 37 °C and 200 rpm for 24 hours. At 0.5 h, 2 h, and 24 h, the suspensions were filtered, and the drug concentration in the filtrate was analyzed by HPLC. The pH of the filtrate was measured at each time point. The solid remaining after 24 hours was identified by XRPD.

[0187] Both forms showed pH-dependent solubility curves, with slightly higher solubility at low pH. At 24 hours, Form A had a higher solubility in SGF than Form B. In the other media, the solubilities of the two forms were similar. There was no change in the XRPD pattern of the solid remaining after 24 hours. The results are shown in Table 14.

[0188] (Table 14) Solubility results of Form A (Pattern 1) and Form B (Pattern 2) in biologically relevant media TIFF0007710449000024.tif57170

[0189] Solid stability of Form B: Approximately 10 mg of Form B was left at 40 °C / 75% RH and 60 °C open for 7 days, and at 25 °C / 92.5% RH open for 10 days. Duplicate samples were prepared under each condition (n = 2). The solid was analyzed by XRPD and HPLC (only for the 40 °C / 75% RH and 60 °C conditions).

[0190] The solid stability tests were performed at 40 °C / 75% RH and 60 °C for 7 days, and at 25 °C / 92.5% RH for 10 days, respectively. The results are summarized in Table 15. Form B was physically stable under the test conditions, with no change in crystal form. Form B was chemically stable at 40 °C / 75% RH for 7 days, but slight degradation was observed at 60 °C for 7 days, with a 0.16% decrease in purity and two new impurities detected (RRT 0.86 and RRT 0.93).

[0191] (Table 15) Stability evaluation results TIFF0007710449000025.tif22146

[0192] Approximately 10 - 20 mg of Compound 2 in Form A and Form B were each manually triturated for 5 minutes. After trituration, there was no change in Form A, but the crystallinity of Form B became weaker. After 5 minutes of trituration, DSC of Form B showed the appearance of Form A after trituration, indicating that there were potential changes in crystal forms during the mechanical treatment process.

[0193] As described above, the solubilities of both Form A and Form B showed pH - dependence. The highest solubility of Form B was 0.056 mg / mL in SGF and the lowest was 0.018 mg / mL in FaSSIF. In SGF, the solubility of Form A was slightly higher than that of Form B, and it was similar in other media. From the solid - state stability results, Form B was physically and chemically stable for 7 days under the conditions of 40 °C / 75% RH, and there were no changes even under the conditions of 92.5% RH for 10 days and 60 °C for 7 days for this crystal form.

[0194] It should be understood that the "Mode for Carrying Out the Invention" section, rather than the "Summary of the Invention" and "Abstract" sections, is used to explain the scope of the claims. The summary and abstract sections can show one or more exemplary embodiments of the invention considered by the inventor, but not all exemplary embodiments, and are not intended to limit the present invention and the appended claims in any way.

[0195] As described above, the present invention has been described using functional configuration blocks for explaining the realization of specific functions and their relationships. For the convenience of explanation, the boundaries of these functional members have been arbitrarily defined in this manuscript. Other boundaries can be defined if the specific functions and their relationships are appropriately implemented.

[0196] Regarding the aspects of the present invention described as genera, each individual type is considered an individual aspect of the present invention. When each aspect of the present invention is described as "comprising features", embodiments "consisting of features" or "essentially consisting of features" are also conceivable.

[0197] The description of the specific embodiments described above sufficiently reveals the general nature of the present invention. Thus, without departing from the general concept of the present invention, others can easily modify and / or adapt various applications such as these specific embodiments by applying the knowledge in the art without undue experimentation. Therefore, based on the teachings and guidance proposed herein, such adaptations and modifications are also included within the meaning and scope of equivalent forms of the disclosed embodiments. It should be understood that the phrases or terms in this specification are for explanatory purposes and not for purposes of limitation. Thus, the terms or phrases in this specification are interpreted by those skilled in the art under the teachings and guidance.

[0198] The breadth and scope of the present invention should not be limited by the above exemplary embodiments.

[0199] The various aspects, embodiments, and options described in this specification can be combined in any variation and all variations.

[0200] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference as if each individual publication, patent, or patent application was specifically and individually incorporated by reference. If the meaning or definition of a term in this specification conflicts with the meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to the term in this specification shall govern.

Claims

1. An amorphous of a compound having the following formula:

2. A crystal of a compound having the following formula: characterized by an X-ray powder diffraction (XRPD) pattern having six or more of the following peaks: 6.2, 12.6, 13.8, 14.8, 15.1, 18.0, 19.6, and 19.9° 2θ ± 0.2°.

3. A crystal of a compound having the following formula: characterized by an X-ray powder diffraction (XRPD) pattern having six or more of the following peaks: 6.2, 12.6, 13.6, 14.5, 14.8, 17.8, 19.0, 19.8, 21.4, 26.3, 31.9 and 38.6° 2θ ± 0.2°.

4. A crystal of a compound having the following formula: characterized by an X-ray powder diffraction (XRPD) pattern having eight or more of the following peaks: 6.2, 6.8, 7.3, 12.5, 14.2, 14.7, 15.7, 16.3, 19.9, 21.2, 22.9 and 26.1° 2θ ± 0.2°.

5. A crystal of a compound having the following formula: characterized by an X-ray powder diffraction (XRPD) pattern having six or more of the following peaks: 5.6, 11.2, 15.8, 16.1, 16.9, 21.4, 22.6 and 34.3° 2θ ± 0.2°.

6. A pharmaceutical composition comprising the amorphous according to Claim 1 or the crystal according to any one of Claims 2 to 5, and optionally a pharmaceutically acceptable excipient.

7. A pharmaceutical composition for treating cancer in a subject, comprising a therapeutically effective amount of the amorphous according to Claim 1 or the crystal according to any one of Claims 2 to 5.

8. The pharmaceutical composition according to Claim 7, wherein the cancer is a hematological malignancy, lung cancer, pancreatic cancer, endometrial cancer, gallbladder cancer, thyroid cancer, cholangiocarcinoma, and / or rectal cancer, and optionally the subject has a G12C mutation in KRAS, HRAS, and / or NRAS.

9. The pharmaceutical composition according to Claim 8, wherein the lung cancer is non-small cell lung cancer. ​

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