Crystalline polymorphs as protein kinase Mek inhibitors, and methods for their preparation and use - Patents.com

Abstract

The present invention relates to crystalline polymorphs of benzothiazole compounds that are protein kinase Mek inhibitors, their preparation methods, and their pharmaceutical uses.

Description

[Technical Field]

[0001] The present invention relates to crystalline polymorphs of benzothiazole compounds that are protein kinase Mek inhibitors, their preparation methods, and their pharmaceutical uses. [Background technology]

[0002] Overactivation of the Ras / Raf / Mek / Erk signaling pathway plays an important role in the proliferation and differentiation of cancer cells. Persistent or overactivation of the Ras / Raf / Mek / Erk signaling pathway has been observed in many cancers, including pancreatic, colon, lung, bladder, kidney, skin, and breast cancer. Inhibition of the Ras / Raf / Mek / Erk signaling pathway is useful for treating such hyperproliferative diseases. Mek, a downstream target of Ras and Raf, plays a key role in this pathway, and the substrate for Mek phosphorylation is the MAP kinase Erk. Inhibition of Mek blocks the Ras / Raf / Mek / Erk signaling pathway, thereby inhibiting cancer cell proliferation. Therefore, Mek inhibitors can inhibit cancer cell proliferation, particularly in cancers caused by overactivation of Ras or Raf. Mek is also involved in inflammatory diseases and conditions, including acute and chronic inflammation.

[0003] Chinese Patent Application No. 201210190520.4 discloses a number of benzothiazole compounds, which exhibit protein kinase Mek inhibitory activity, including the compound 4-fluoro-5-(2-fluoro-4-iodophenylamino)-1H-benzo[d]thiazole-6-carboxylic acid (2-hydroxyethoxy)-amide (hereinafter referred to as Compound 1). However, according to the preparation method of Chinese Patent Application No. 201210190520.4, the solid form of Compound 1 is amorphous. However, the purity of the amorphous form is usually difficult to control, and the physical and chemical stability is usually poor, with low hygroscopicity. Furthermore, Compound 1 is unstable to light irradiation and decomposes to produce impurities (A). [ka] This tends to produce hydroxybenzoates, which affects the efficacy and storage stability of Compound 1 itself.

[0004] Therefore, it is necessary to develop a new crystalline form of the compound that is advantageous in improving purity control in process production, and in improving the stability and storage durability of the compound. Summary of the Invention

[0005] The present invention provides crystalline polymorphs of Compound 1, which have advantages such as high purity, good solid-state stability, good powder properties, and mechanical stability. More specifically, the present invention provides seven crystalline forms of Compound 1, namely, Form I, Form II, Form IIIA, Form IIIB, Form IV, Form V, and Form VI, in which Form I and Form IIIA exhibit superior properties such as solid-state stability, mechanical stability, hygroscopicity, and light stability compared to other crystalline forms or amorphous forms, and are therefore advantageous for pharmaceutical manufacturing.

[0006] Specifically, the Type I crystal and Type IIIA crystal of Compound 1 exhibit one or more of the following advantages over the amorphous form: 1. The results of the suspension competition showed that type I crystals are thermodynamically more stable than type IIIA crystals in the range of room temperature to 50°C. 2. The DVS results showed that the Type I crystal and Type IIIA crystal were almost non-hygroscopic and no crystal form conversion occurred after the DVS test, while the amorphous form was converted to Type I crystal after the DVS test. 3. The results of solid state stability showed that the I-type crystal did not undergo crystal form conversion or purity loss after being left sealed at 60°C for 1 day and then left open at 25°C / 60%RH and 40°C / 75%RH for 1 week, indicating that the I-type crystal has good physical and chemical stability under the evaluation conditions. The IIIA-type crystal showed no change in purity after 1 day at 60°C, but the I-type crystal diffraction peak was observed, indicating that the I-type crystal did not undergo crystal form conversion or purity loss after 1 week at 25°C / 60%RH and 40°C / 75%RH. The amorphous material showed no obvious change in purity under the three evaluation conditions, but all of the crystal forms were converted to the I-type crystal. 4. The results of the light irradiation stability showed that the I-type crystal was stable under light irradiation (white light 5890 Lux + ultraviolet light 8.7 W / m 2 ) conditions for 24 hours, the purity was clearly higher than that of the IIIA type crystals, the V type crystals and the amorphous form, and the photostability was superior. 5. The results of the dynamic solubility test of the Form I crystal and the amorphous substance in 1M HCl and pH 1.0 / 2.0 / 4.5 / 7.4 buffers at room temperature showed that the solubility of the amorphous substance in all media within 10 minutes was higher than that of the Form I crystal, and all showed a process of first increasing and then decreasing, with the highest solubility measured in 1M HCl being higher than in other pH buffers. Compared with the amorphous substance, the Form I crystal dissolves more slowly in each medium (different pH values), and its solubility in 1M HCl is slightly higher than in other pH buffers, which ensures more stable dissolution of the Form I crystal in vivo and makes it easier to achieve stable in vivo pharmacokinetics with stable blood drug concentration, which is advantageous for avoiding the risk of medication safety due to excessive fluctuations in blood drug concentration. 6. The powder property test results show that the Type I crystalline and amorphous samples have similar fluidity, and the mechanical stability results show that after tableting (350 MPa) and manual crushing (about 3 minutes), no crystal form transformation occurred in the Type I crystalline and the crystallinity did not decrease significantly, and the amorphous was transformed into the Type I crystalline, and no crystal form transformation occurred in the Type IIIA crystalline, but the crystallinity decreased; and / or 7. In pharmacokinetic experiments, the type I crystals showed higher exposure and blood drug concentrations than the type IIIA crystals, suggesting the possibility of reducing the dosage.

[0007] Based on the characterization data and evaluation results, crystalline form I exhibited superior photostability and no crystal form conversion occurred under all evaluation conditions, whereas the amorphous form converted to crystalline form I after DVS, solid state stability, solubility, and mechanical stability tests.

[0008] In a first aspect, the present invention provides a crystalline polymorph of formula (I): [ka] Here, n is 0 or 1, and X is acetonitrile, water, 1,4-dioxane, ethanol, methanol, dimethylformamide, acetone, or a mixture thereof.

[0009] In some embodiments, n is 0. In some embodiments, n is 1 and X is acetonitrile, water, 1,4-dioxane, ethanol, or dimethylformamide.

[0010] In some embodiments, n is 0, the crystalline polymorph is a Form I crystal, and the X-ray powder diffraction pattern of the Form I crystal has characteristic diffraction peaks at 2θ positions of 16.71°±0.2°, 21.82°±0.2°, and 23.75°±0.2°, using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at 2θ positions of 7.48°±0.2° and 22.36°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at 2θ positions of 5.3°±0.2°, 24.57°±0.2°, and 27.08°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at the 2θ positions of 11.81°±0.2°, 15.83°±0.2°, 17.92°±0.2°, 18.95°±0.2°, and 19.17°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at the 2θ positions of 5.30°±0.2°, 7.48°±0.2°, 11.81°±0.2°, 14.85°±0.2°, 15.83°±0.2°, 16.71°±0.2°, 17.92°±0.2°, 18.95°±0.2°, 19.17°±0.2°, 19.43°±0.2°, 21.14°±0.2°, 21.82°±0.2°, 22. The Form I crystal has characteristic diffraction peaks at the following 2θ positions: 23.75°±0.2°, 24.57°±0.2°, 27.08°±0.2°, 27.83°±0.2°, 28.88°±0.2°, 31.20°±0.2°, 31.92°±0.2°, 32.40°±0.2°, 33.91°±0.2°, 35.83°±0.2°, 37.51°±0.2°, and 39.04°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form I crystal is essentially as shown in FIG. 3. In some embodiments, the Form I crystal has a TGA pattern and / or a DSC pattern as shown in FIG. 4. In some embodiments, the Form I crystal is an anhydrate.

[0011] In some embodiments, n is 1, X is acetonitrile, the crystalline polymorph is Form II crystal, and the X-ray powder diffraction pattern of the Form II crystal has characteristic diffraction peaks at 2θ positions of 24.99°±0.2°, 26.05°±0.2°, and 22.6°±0.2° using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Form II crystal also has characteristic diffraction peaks at 2θ positions of 6.35°±0.2°, 20.34°±0.2°, 22.41°±0.2°, and 28.71°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form II crystal also has characteristic diffraction peaks at the following 2θ positions: 9.18°±0.2°, 16.03°±0.2°, 18.25°±0.2°, 27.07°±0.2°, 29.08°±0.2°, and 33.93°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form II crystal also has characteristic diffraction peaks at the following 2θ positions: 14.44°±0.2°, 24.64°±0.2°, 26.41°±0.2°, 32.27°±0.2°, 32.68°±0.2°, 37.07°±0.2°, and 39.51°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form II crystal is 6.35°±0.2°, 9.18°±0.2°, 9.91°±0.2°, 14.44°±0.2°, 16.03°±0.2°, 18.25°±0.2°, 19.77°±0.2°, 20.34°±0.2°, 21.81°±0.2°, 22.41°±0.2°, 22.60°±0.2°, 23.84°±0.2°, 24.64°±0.2°, 24.99°±0.2°, 25.43°±0.2°, 26.05°±0.2° 0.2°, 26.41°±0.2°, 27.07°±0.2°, 28.71°±0.2°, 29.08°±0.2°, 29.81°±0.2°, 31.16°±0.2°, 31.57°±0.2°, 32.27°±0.2°, 32.68°±0.2°, 33.93°±0.2°, 34.19°±0.2°, 35.42°±0.2°, 37.07°±0.2°, 37.56°±0.2°, 38.69°±0.2°, and 39.51°±0.2° 2θ positions. In some embodiments, the X-ray powder diffraction pattern of the Form II crystal is essentially as shown in FIG.In some embodiments, the Form II crystal has a TGA pattern and / or a DSC pattern shown in FIG.

[0012] In some embodiments, n is 0, the crystalline polymorph is a Form IIIA crystal, and the X-ray powder diffraction pattern of the Form IIIA crystal has characteristic diffraction peaks at 2θ positions of 6.59°±0.2°, 22.69°±0.2°, 20.32°±0.2°, 23.62°±0.2°, 23.91°±0.2°, and 24.15°±0.2°, using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Form IIIA crystal also has characteristic diffraction peaks at 2θ positions of 10.8°±0.2°, 17.14°±0.2°, 13.75°±0.2°, 21.59°±0.2°, and 26.01°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IIIA crystal also has characteristic diffraction peaks at the following 2θ positions: 18.71°±0.2°, 21.97°±0.2°, 25.54°±0.2°, 27.13°±0.2°, 27.59°±0.2°, and 30.51°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IIIA crystal is 6.59°±0.2°, 9.9°±0.2°, 10.8°±0.2°, 13.09°±0.2°, 13.75°±0.2°, 17.14°±0.2°, 17.87°±0.2°, 18.71°±0.2°, 19.19°±0.2°, 20.32°±0.2°, 21.59°±0.2°, 21.97°±0.2°, 22.69°±0.2°, 23.62°±0.2°, 23.91°± The Form IIIA crystal has characteristic diffraction peaks at the following 2θ positions: 24.15°±0.2°, 25.54°±0.2°, 26.01°±0.2°, 27.13°±0.2°, 27.59°±0.2°, 28.83°±0.2°, 29.24°±0.2°, 30.51°±0.2°, 31.13°±0.2°, 31.79°±0.2°, 33.6°±0.2°, 34.14°±0.2°, 36.08°±0.2°, 36.67°±0.2°, and 37.26°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IIIA crystal is essentially as shown in FIG. 9. In some embodiments, the Form IIIA crystal has a TGA pattern and / or a DSC pattern as shown in FIG. 10. In some embodiments, the Form IIIA crystal is an anhydrate.

[0013] In some embodiments, n is 0, the crystalline polymorph is a Type IIIB crystal, and the X-ray powder diffraction pattern of the Type IIIB crystal has characteristic diffraction peaks at 2θ positions of 6.53°±0.2°, 13.69°±0.2°, 18.6°±0.2°, 20.19°±0.2°, 21.52°±0.2°, and 22.64°±0.2°, using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Type IIIB crystal also has characteristic diffraction peaks at 2θ positions of 10.75°±0.2°, 17.07°±0.2°, 21.93°±0.2°, 26.13°±0.2°, 23.57°±0.2°, and 30.46°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IIIB crystal also has characteristic diffraction peaks at the following 2θ positions: 13.05°±0.2°, 16.63°±0.2°, 20.82°±0.2°, 24.01°±0.2°, 27.55°±0.2°, and 31.79°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IIIB crystals is 6.53°±0.2°, 10.75°±0.2°, 12.62°±0.2°, 13.05°±0.2°, 13.69°±0.2°, 16.63°±0.2°, 17.07°±0.2°, 18.60°±0.2°, 19.59°±0.2°, 20.19°±0.2°, 20.82°±0.2°, 21.52°±0.2°, 21.93°±0.2° 13.57°±0.2°, 24.01°±0.2°, 25.46°±0.2°, 26.13°±0.2°, 27.55°±0.2°, 30.46°±0.2°, 31.04°±0.2°, 31.79°±0.2°, 32.81°±0.2°, 33.54°±0.2°, 34.06°±0.2°, and 34.46°±0.2° 2θ positions. In some embodiments, the X-ray powder diffraction pattern of the Form IIIB crystal is essentially as shown in FIG. 13. In some embodiments, the Form IIIB crystal has a TGA pattern and / or a DSC pattern as shown in FIG. 14.

[0014] In some embodiments, n is 1, X is 1,4-dioxane, the crystalline polymorph is Form IV crystal, and the X-ray powder diffraction pattern of the Form IV crystal has characteristic diffraction peaks at 8.56°±0.2°, 13.29°±0.2°, 17.69°±0.2°, 19.75°±0.2°, and 22.45°±0.2° 2θ positions, characterized by using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Form IV crystal also has characteristic diffraction peaks at 5.26°±0.2°, 18.29°±0.2°, 31.83°±0.2°, 25.68°±0.2°, 22.86°±0.2°, 32.81°±0.2°, and 23.44°±0.2° 2θ positions. In some embodiments, the X-ray powder diffraction pattern of the Form IV crystal also has characteristic diffraction peaks at the following 2θ positions: 126.57°±0.2°, 27.52°±0.2°, 35.69°±0.2°, 21.09°±0.2°, 20.35°±0.2°, and 31.43°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form IV crystal is 5.26°±0.2°, 8.56°±0.2°, 9.85°±0.2°, 13.29°±0.2°, 17.69°±0.2°, 18.29°±0.2°, 19.75°±0.2°, 20.35°±0.2°, 21.09°±0.2°, 22.45°±0.2°, 22.86°±0.2°, 23.44°±0.2° 17.2°±0.2°, 24.44°±0.2°, 25.68°±0.2°, 26.57°±0.2°, 27.52°±0.2°, 28.40°±0.2°, 29.78°±0.2°, 31.43°±0.2°, 31.83°±0.2°, 32.81°±0.2°, 34.29°±0.2°, 35.69°±0.2°, and 37.72°±0.2° 2θ positions. In some embodiments, the Form IV crystal has an X-ray powder diffraction pattern essentially as shown in FIG. 17. In some embodiments, the Form IV crystal has a TGA pattern and / or a DSC pattern as shown in FIG. 18.

[0015] In some embodiments, n is 1, X is ethanol, the crystalline polymorph is Form V crystals, and the X-ray powder diffraction pattern of the Form V crystals has characteristic diffraction peaks at 2θ positions of 6.21°±0.2°, 8.47°±0.2°, 15.62°±0.2°, 21.73°±0.2°, 25.53°±0.2°, 25.94°±0.2°, and 28.05°±0.2°, using Cu-Kα radiation. In some embodiments, the X-ray powder diffraction pattern of the Form V crystals also has characteristic diffraction peaks at 2θ positions of 9.61°±0.2°, 17.55°±0.2°, 19.25°±0.2°, 22.22°±0.2°, 23.12°±0.2°, 32.92°±0.2°, and 34.22°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form V crystals also has characteristic diffraction peaks at the following 2θ positions: 9.06°±0.2°, 20.07°±0.2°, 28.49°±0.2°, 30.21°±0.2°, 31.25°±0.2°, 35.47°±0.2°, and 38.94°±0.2°. In some embodiments, the X-ray powder diffraction pattern of the Form V crystals is 6.21°±0.2°, 8.47°±0.2°, 25.94°±0.2°, 15.62°±0.2°, 25.53°±0.2°, 28.05°±0.2°, 21.73°±0.2°, 17.55°±0.2°, 32.92°±0.2°, 23.12°±0.2°, 22.22°±0.2°, 19.25°±0.2°, 34.22°±0.2°, 9.61°±0.2°, 9.06°±0.2°. 20°±0.2°, 38.94°±0.2°, 31.25°±0.2°, 35.47°±0.2°, 28.49°±0.2°, 30.21°±0.2°, 20.07°±0.2°, 39.78°±0.2°, 14.26°±0.2°, 24.32°±0.2°, 16.95°±0.2°, 32.33°±0.2°, 36.43°±0.2°, 24.94°±0.2°, 12.42°±0.2°, and 37.86°±0.2° 2θ positions. In some embodiments, the X-ray powder diffraction pattern of the Form V crystal is essentially as shown in FIG. 20. In some embodiments, the Form V crystal has a TGA pattern and / or a DSC pattern as shown in FIG. 21.

[0016] In some embodiments, n is 1, X is dimethylformamide, and the crystalline polymorph is Form VI, and in some embodiments, the X-ray powder diffraction pattern of the Form VI crystal is essentially as shown in Figure 23. In some embodiments, the Form VI crystal has a TGA pattern and / or a DSC pattern as shown in Figure 24.

[0017] In some embodiments, the Form I crystals, Form II crystals, Form IIIA crystals, Form IIIB crystals, Form IV crystals, Form V crystals, and Form VI crystals have a purity of about 85% or more, for example, about 90% or more, for example, about 95% or more, for example, about 97% or more, for example, about 99% or more, and about 99.9% or more, based on the weight of Compound 1, as determined by HPLC (high performance liquid chromatography). Other materials may include crystalline forms of Compound 1 and / or reaction impurities and / or processing impurities prepared therefrom, such as the photolytic impurity Compound (A). Mixtures of Form I crystals of Compound 1 with other solid forms (e.g., other crystalline forms and amorphous forms) are also within the scope of the present disclosure.

[0018] In some embodiments, the crystalline polymorph of Compound 1 is essentially free of impurity (A). In some embodiments, the crystalline polymorph of Compound 1 contains less than 0.15% by weight of impurity (A) relative to the crystalline polymorph, e.g., 0.10%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, or 0.01% by weight of impurity (A).

[0019] In some embodiments, the Form I crystals of Compound 1 are essentially free of impurity (A). In some embodiments, the Form I crystals of Compound 1 contain less than 0.15% by weight of impurity (A) relative to the Form I crystals, for example, 0.10%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, or 0.01% by weight of impurity (A).

[0020] In some embodiments, the Form I crystal, the Form II crystal, the Form IIIA crystal, the Form IIIB crystal, the Form IV crystal, the Form V crystal, and the Form VI crystal each have a crystallinity of about 85% or more, such as about 90% or more, for example about 95% or more, such as about 97% or more, for example about 99% or more, and including about 99.9% or more.

[0021] In a second aspect, the present invention provides a pharmaceutical composition comprising any one of the crystalline polymorphs of the Form I crystals to Form VI crystals of the present invention and a pharmaceutically acceptable carrier and / or excipient. In some embodiments, the pharmaceutical composition comprises the Form I crystals or Form IIIA crystals of the present invention and a pharmaceutically acceptable carrier and / or excipient. In some embodiments, the pharmaceutical composition comprises the Form I crystals of the present invention and a pharmaceutically acceptable carrier and / or excipient.

[0022] In a third aspect, the present invention provides a method for treating a mammalian tumor, a chronic inflammatory disease, an inflammatory bowel disease, a skin disease, diabetes, an eye disease, a disease associated with angiogenesis or revascularization in a mammal, a disease associated with chronic pain, and other diseases regulated by the Mek cascade, comprising administering to the mammal a crystalline polymorph of any one of the Forms I to VI of the present invention. The present invention also provides a crystalline polymorph of any one of the Forms I to VI of the present invention for treating a mammalian tumor, a chronic inflammatory disease, an inflammatory bowel disease, a skin disease, diabetes, an eye disease, a disease associated with angiogenesis or revascularization in a mammal, a disease associated with chronic pain, and other diseases regulated by the Mek cascade. The present invention also provides a use of a crystalline polymorph of any one of the Forms I to VI of the present invention in a preparation for treating a mammalian tumor, a chronic inflammatory disease, an inflammatory bowel disease, a skin disease, diabetes, an eye disease, a disease associated with angiogenesis or revascularization in a mammal, a disease associated with chronic pain, and other diseases regulated by the Mek cascade. In some embodiments, the mammal is a human.

[0023] In a fourth aspect, the present invention provides a method for treating a mammalian RAS or RAF mutant cancer, the method comprising administering 4-fluoro-5-(2-fluoro-4-iodophenylamino)-1H-benzo[d]thiazole-6-carboxylic acid (2-hydroxyethoxy)-amide (Compound 1) or a pharmaceutically acceptable salt thereof to the mammal. In some embodiments, Compound 1 is in any one of crystalline polymorphs selected from Forms I to VI. In some embodiments, the RAS or RAF mutant cancer is, for example, a KRAS mutant cancer, an NRAS mutant cancer, an HRAS mutant cancer, or a BRAF mutant cancer. In some embodiments, the RAS mutant cancer is pancreatic cancer, colorectal cancer, lung cancer, melanoma, acute myeloid leukemia, bladder cancer, or head and neck cancer. In a preferred embodiment, the cancer is an NRAS mutant cancer. In some embodiments, the NRAS mutant cancer is an NRAS mutant melanoma.

[0024] In one embodiment, KRAS comprises a mutation at one or more positions selected from codons 12, 13, 59, and 61. In one embodiment, the KRAS mutant form has a mutation at one or more amino acid positions selected from G12, G13, S17, P34, A59, and Q61. In one embodiment, the KRAS mutant form has one or more amino acid substitutions selected from G12C, G12S, G12R, G12F, G12L, G12N, G12A, G12D, G12V, G13C, G13S, G13D, G13V, G13P, S17G, P34S, A59E, A59G, A59T, Q61K, Q61L, Q61R, and Q61H. In one embodiment, the KRAS mutant form has a mutation at one or more amino acid positions selected from G12, G13, A59, Q61, K117, and A146. In one embodiment, the KRAS mutant form has one or more amino acid substitutions selected from G12C, G12R, G12S, G12A, G12D, G12V, G13C, G13R, G13S, G13A, G13D, G13V, A59E, A59G, A59T, Q61K, Q61L, Q61R, Q61H, K117N, K117R, K117E, A146P, A146T, and A146V. In one embodiment, the BRAF mutation is a BRAF V600E mutation.

[0025] In one embodiment, NRAS comprises a mutation at one or more positions selected from codons 12, 13, 59, 61, and 146. In some embodiments, the NRAS mutant form has a mutation at one or more amino acid positions selected from G12, G13, A59, Q61, K117, and A146. In some embodiments, the NRAS mutant form has one or more amino acid substitutions selected from G12C, G12R, G12S, G12A, G12D, G12V, G13C, G13R, G13S, G13A, G13D, G13V, A59D, A59T, Q61K, Q61L, Q61R, Q61H, K117N, K117R, K117E, A146P, A146T, and A146V.

[0026] In some embodiments, the cancer is early-stage, intermediate-stage, or advanced-stage cancer. The cancer may be locally advanced or metastatic. In some embodiments, the mammal has previously undergone immunotherapy. In some embodiments, the mammal has previously undergone immunotherapy and is afflicted with NRAS-mutated advanced melanoma. In some embodiments, the melanoma is selected from advanced melanoma, unresectable melanoma, metastatic melanoma, BRAF-mutated melanoma, NRAS-mutated melanoma, cutaneous melanoma, or intraocular melanoma.

[0027] In some embodiments, Compound 1 is in the form of a tablet, powder, granule, patch, inhalant, or capsule. In some embodiments, Compound 1 is in the form of a capsule. In some embodiments, Compound 1 is administered once or twice daily at a dose of 5 to 50 mg. In some embodiments, Compound 1 is administered twice daily at a dose of 12 mg.

[0028] In a fifth aspect, the present invention provides a method for preparing a Form I crystal of the present invention, said method comprising: a) adding an amorphous sample of Compound 1 to a solvent, then heating to a temperature of about 70° C. or higher, allowing the resulting clear solution to cool to room temperature, keeping it at room temperature and stirring continuously, allowing the solid to precipitate, filter and dry; or b) dissolving an amorphous sample of Compound 1 in a good solvent, filtering to obtain a clear solution, and adding a poor solvent while stirring the clear solution until a solid precipitates; or c) dissolving an amorphous sample of Compound 1 in a solvent, stirring at about 50°C, filtering to obtain a filtrate, cooling the resulting filtrate to about 5°C, and collecting the precipitated solid.

[0029] In some embodiments, the heating temperature in method 1) is about 75°C to about 100°C, about 80°C to about 90°C, or about 75°C or about 85°C. In some embodiments, the room temperature in method 1) is about 20 to about 25°C. In some embodiments, the continuous stirring time in method 1) is about 0.5 to 12 hours, about 1 to 12 hours, about 1 to 8 hours, about 1 to 5 hours or more, or the continuous stirring time is about 24 to about 96 hours. In some embodiments, the clear solution in method 1) is cooled to room temperature within about 2 to about 5 hours or within about 2.5 to about 3 hours. In some embodiments, after maintaining the temperature and stirring at room temperature in method 1), the temperature can be optionally further lowered to about 0 to about 10°C and the mixture can be stirred while maintaining the temperature. In some embodiments, the mixture is incubated at about 0 to about 10°C and stirred for about 1 to about 12 hours, about 1 to about 8 hours, or about 1 to about 5 hours, or optionally incubated at about 0 to about 10°C and stirred for longer periods. In some embodiments, the solvent in method 1) is water, methanol, ethanol, isopropyl alcohol, acetone, methyl isobutyl ketone, 2-butanone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, tetrahydrofuran, anisole, 2-methyltetrahydrofuran, cyclopentyl methyl ether, 1,4-dioxane, acetonitrile, dichloromethane, toluene, meta-xylene, n-heptane, n-hexane, n-pentane, dimethyl sulfoxide, dimethylacetamide, N-methylpyrrolidone, or a mixture thereof. In some embodiments, the solvent in method 1) is ethanol.

[0030] In some embodiments, the good solvent in method 2) is a solvent in which compound 1 can be dissolved, and the poor solvent in method 2) is a solvent in which compound 1 cannot be dissolved. In some embodiments, the good solvent in method 2) is MEK, 1,4-dioxane, or DMSO. In some embodiments, the poor solvent in method 2) is MTBE, EtOAc, CHCl, n-heptane, Anisole, EtOAc, HO, IPAc, CPME, DCM, or toluene.

[0031] In some embodiments, the cooling rate in method 3) is about 0.1° C. / min. In some embodiments, the solvent in method 3) is MIBK, methyl acetate, 2-MeTHF, or acetone / EtOH (1:1).

[0032] The present invention provides a method for preparing the Type V crystal of the present invention, the method comprising adding an amorphous sample of Compound 1 to a solvent, heating at a temperature of about 70°C or less, cooling the resulting clear solution to about 0 to about 10°C, and allowing it to stand at about 5°C until a solid precipitates, followed by drying. In some embodiments, the method includes heating at about 70°C, about 60°C, or about 50°C. In some embodiments, the cooling rate is about 0.1 to about 0.5°C / min. In some embodiments, the cooling rate is about 0.1°C / min. In some embodiments, the cooling rate is about 0.1°C / min. In some embodiments, the resulting clear solution is cooled to about 5°C.

[0033] The present invention provides a method for preparing a Form IIIA crystal of the present invention, the method comprising heating a Form II crystal to a first temperature, incubating it for about 3 to about 10 minutes, and then cooling it to obtain a Form IIIA crystal. In some embodiments, the first temperature is about 100 to about 140°C, e.g., about 110 to about 130°C, e.g., about 120°C. In some embodiments, the Form II crystal is incubate at the first temperature for about 5 minutes. In some embodiments, after incubating the Form II crystal, it is cooled to room temperature to about 50°C, e.g., cooled to room temperature.

[0034] Differential scanning calorimetry (DSC) is well known in the art, and while the height of the melting peak in a DSC curve depends on many factors related to sample preparation and instrument geometry, the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystalline compounds of the present invention have a DSC pattern with characteristic peak positions and substantially the same properties as the DSC patterns shown in the drawings of the present invention, and the measurement error tolerance is required to be within ±5°C, typically within ±3°C.

[0035] The numerical values ​​described and claimed in this invention are approximate values. Variations within the numerical values ​​may be due to instrument calibration, instrument error, crystal purity, crystal size, sample size, and other factors.

[0036] The crystalline forms of the present invention are not limited to those having characteristic spectra, such as XRPD, DSC, TGA, DVS, and isothermal adsorption curves, that are exactly the same as those depicted in the drawings disclosed in the present invention, and any crystalline form having characteristic spectra that are basically the same or essentially the same as those depicted in the drawings is included within the scope of the present invention.

[0037] term Unless specifically defined otherwise herein, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art.

[0038] Unless the context clearly dictates otherwise, this application includes singular terms such as "a," "an," and "said" as used in the claims, and includes their corresponding plural forms. So, for example, reference to "a crystalline form" includes one or more of such different crystalline forms, and reference to "the process" includes reference to equivalent steps and methods known to those skilled in the art that may improve upon or substitute for the method described herein.

[0039] Throughout the following specification and claims, unless the context requires otherwise, the term "comprising," and variations such as "containing" and "including," will be understood to implicitly include the integer or step or set of integers or steps, but not to exclude any other integer or step or set of integers or steps. When used in this application, the term "comprising" can be replaced with the term "containing," or when used, with the term "having."

[0040] The term "about" means plus or minus 10%, 5%, or 2% of the value of the specified item.

[0041] A "therapeutically effective amount" refers to an amount of a compound that elicits a physiological or medical response in a tissue, system, or subject, and includes an amount of compound sufficient to prevent or alleviate to some extent one or more symptoms of the disease or condition being treated when administered to a subject, as desired. A "therapeutically effective amount" can vary depending on the compound, the disease, condition, and / or symptoms of the disease or condition, the severity of the disease or condition, and / or symptoms of the disease or condition, the age of the subject being treated, and / or the weight of the subject being treated. The appropriate amount in any particular case will be apparent to one of ordinary skill in the art or can be determined by routine experimentation. In the case of combination therapy, a "therapeutically effective amount" refers to the total amount of the combined compounds effective to treat the disease, condition, or condition.

[0042] An "excipient" refers to something that is not itself a therapeutic agent, but is used as a diluent, adjuvant, adhesive and / or vehicle, added to a pharmaceutical composition to improve its handling or storage properties, or to enable or facilitate the compound or pharmaceutical composition to form into a unit dosage form for administration.

[0043] "Crystalline form" or "crystal" or "crystalline polymorph" refers to any solid material that exhibits three-dimensional arrangement and, in contrast to amorphous solid materials, displays a characteristic XRPD pattern of clearly defined peaks.

[0044] "Amorphous" refers to a solid form of non-crystalline molecules and / or ions. Amorphous solids do not exhibit X-ray diffraction patterns with sharp maxima.

[0045] "Hydrate" refers to a crystalline form of a molecule that further contains water incorporated into the crystalline structure. The water molecules in a hydrate can be in an ordered and / or disordered arrangement. Hydrates may contain stoichiometric or non-stoichiometric amounts of water molecules.

[0046] "Anhydrate" refers to a crystalline form that is essentially free of water molecules in any form in the crystal, e.g., the crystal lattice or unit cell of the crystal is essentially free of water molecules.

[0047] The term "solvate" refers to a crystalline form of a molecule, which further comprises one or more solvent molecules incorporated into the crystalline structure. The solvent molecules in a solvate can be in an ordered and / or disordered arrangement. A solvate may contain stoichiometric or non-stoichiometric amounts of solvent molecules. Exemplary solvates include, but are not limited to, hydrates, ethanol salts, methanol salts, and isopropyl alcohol salts, acetic acid salts. Solvation methods are generally known in the art. It should be noted that in solvates, the substance bound to the host molecule (e.g., active drug ingredient) is liquid at room temperature, whereas in cocrystals, the substance is solid at room temperature.

[0048] The crystalline forms disclosed herein are essentially pure crystals. As used herein, the term "essentially pure" refers to at least 85% by weight, preferably at least 95% by weight, more preferably at least 99% by weight, of the crystalline forms disclosed herein, and further includes about 100% by weight of a particular crystalline form. Other materials include one or more other forms of the compound and / or reaction and / or processing impurities prepared therefrom. For example, a crystalline form of Compound 1 may be considered essentially pure because it has a purity of greater than 90% by weight, as measured by means then known and generally accepted in the art, where less than 10% by weight of other materials include amorphous Compound 1 and / or one or more other forms and / or reaction and / or processing impurities.

[0049] "X-ray powder diffraction pattern (XRPD pattern)" refers to an experimentally observed diffraction pattern or parameters, data, or values ​​derived therefrom. XRPD patterns are typically characterized by peak positions (abscissa) and / or peak intensities (ordinate). For the crystalline forms disclosed herein, only the main peaks (i.e., the most characteristic, prominent, particular, and / or reproducible peaks) are summarized; other peaks can be obtained from the diffraction pattern by conventional methods. The main peaks can be reproduced within a margin of error (the last given decimal point ±2, or the given value ±0.2).

[0050] "2θ" refers to the peak position, expressed in degrees (°), established based on an X-ray diffraction experiment, and is usually the abscissa unit in a diffraction pattern. If a reflection is diffracted when the incident beam forms an angle θ with a lattice plane, the experimental setup requires recording the reflected beam at an angle 2θ. It should be understood that the specific 2θ values ​​of specific crystalline forms referred to herein are intended to refer to the 2θ values ​​(expressed in degrees) measured using the X-ray diffraction experimental conditions described herein.

[0051] The terms "essentially the same" or "essentially as shown in Figure XX" with respect to X-ray diffraction peaks are intended to take into account variations in typical peak positions and intensities. For example, those skilled in the art will understand that peak positions (2θ) will exhibit some degree of variation, typically amounting to 0.1-0.2°, and that some variation will occur depending on the instrument used to measure the diffraction. Those skilled in the art will also understand that relative peak intensities will vary depending on instrumental differences and on the degree of crystallinity, preferred orientation, sample surface preparation, and other factors known to those skilled in the art, and should be considered only as qualitative measurements.

[0052] Pharmaceutical compositions containing the compounds disclosed herein can be administered orally, inhaled, rectally, parenterally, or topically to a subject in need thereof. For oral administration, the pharmaceutical composition may be in the form of a regular solid formulation such as a tablet, powder, granules, or capsule, or a liquid formulation such as a water or oil suspension, or other liquid formulation such as a syrup, solution, or suspension. For parenteral administration, the pharmaceutical composition may be in the form of a solution, aqueous solution, oil suspension concentrate, lyophilized powder, or the like. Preferably, the pharmaceutical composition is selected from the group consisting of a tablet, coated tablet, capsule, suppository, nasal spray, or injection, and more preferably a tablet or capsule. The pharmaceutical composition can be administered as a single unit at a precise dose. The pharmaceutical composition may also contain additional active ingredients.

[0053] All formulations of the pharmaceutical compositions disclosed herein can be prepared by conventional methods in the pharmaceutical field. For example, the desired formulation can be prepared by mixing the active ingredient with one or more excipients. "Pharmaceutically acceptable excipients" refer to conventional drug carriers used in the desired drug formulation, such as diluents, vehicles such as water and various organic solvents, fillers such as starch and sucrose, adhesives such as cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone (PVP), humectants such as glycerol, disintegrants such as agar, calcium carbonate, and sodium bicarbonate, absorption enhancers such as quaternary ammonium compounds, surfactants such as cetyl alcohol, absorbent carriers such as kaolin and bentonite, and lubricants such as talc, calcium stearate, magnesium stearate, and polyethylene glycol. Pharmaceutical compositions may also contain other pharmaceutically acceptable excipients, such as decentralized agents, stabilizers, thickeners, complexing agents, buffers, penetration enhancers, polymers, aromatic compounds, sweeteners, and dyes.

[0054] A "pharmaceutical composition" refers to a composition comprising a crystalline form of a compound of the present invention in combination with at least one additional pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. A "pharmaceutically acceptable carrier" refers to a vehicle generally accepted in the art for delivering a bioactive agent to an animal, particularly a mammal, and includes, depending on the mode of administration and the nature of the dosage form, adjuvants, excipients, or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, and dispensing agents.

[0055] Pharmaceutically acceptable carriers are prepared based on many factors within the knowledge of those skilled in the art. These include, but are not limited to, the type and nature of the active agent being prepared, the subject to whom the composition containing the agent will be administered, the intended route of administration of the composition, and the targeted therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as a variety of solid and semi-solid dosage forms. Such carriers may further contain many different components and additives other than the active agent, and such additional components are included in the formulation for a variety of reasons well known to those skilled in the art (e.g., stabilization of the active agent, adhesives, etc.). Descriptions of suitable pharmaceutically acceptable carriers and factors related to their selection can be found in a variety of readily available sources, such as Allen, Jr., LV et al., Remington: The Science and Practice of Pharmacy (Vol. 2), 22nd Edition, Pharmaceutical Press (2012).

[0056] Naturally, the dosage regimen for the solid forms of the present application will vary depending on known factors, such as the pharmacological properties of the particular drug and its mode and route of administration, the recipient's species, age, sex, health status, treatment status, and weight, the nature and severity of symptoms, the type of concurrent treatment, the frequency of treatment, the route of administration, the patient's renal and hepatic function, and the desired effect. Typically, the daily oral dose range for each active ingredient is about 0.001 to about 5000 mg / day, preferably about 0.01 to about 1000 mg / day, and most preferably 0.1 to about 250 mg / day. For intravenous administration, the most preferred dose range is about 0.01 to about 10 mg / kg / minute during a constant rate infusion. The compounds of the present invention may be administered in a single daily dose, or the total daily dose may be administered in divided doses two, three, or four times daily.

[0057] Dosage forms (pharmaceutical compositions) for administration may contain from about 1 mg to about 2000 mg of the active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient is typically present in an amount of from about 0.1% to 95% by weight, based on the total weight of the composition. [Brief explanation of the drawings]

[0058] [Figure 1] 1 is an XRPD pattern of amorphous Compound 1. [Figure 2] Amorphous dynamic water sorption (DVS) of Compound 1. [Figure 3] 1 is an XRPD pattern of the type I crystal of Example 2a. [Figure 4] 1 is a TGA / DSC pattern of the I-type crystal of Example 2a. [Figure 5] 1 is a 1H NMR spectrum of the type I crystal of Example 2a. [Figure 6] 1 shows XRPD patterns of the Type II crystals of Examples 3a and 3b. [Figure 7] 1 is a TGA / DSC pattern of the type II crystal of Example 3a. [Figure 8] 1 is a 1H NMR spectrum of the type II crystal of Example 3a. [Figure 9]1 is an XRPD pattern of the IIIA type crystal of Example 4a. [Figure 10] 1 is a TGA / DSC pattern of the IIIA type crystal of Example 4a. [Figure 11] 1 is a 1H NMR spectrum of the IIIA type crystal of Example 4a. [Figure 12] 1 shows XRPD overlay patterns of the IIIA crystal of Example 4a before and after heating. [Figure 13] 1 is an XRPD pattern of the IIIB type crystal of Example 4b. [Figure 14] 1 is a TGA / DSC pattern of the IIIB type crystal of Example 4b. [Figure 15] 1 is a 1H NMR spectrum of the IIIB type crystal of Example 4b. [Figure 16] 1 shows an XRPD pattern of the type IIIB crystal of Example 4b as a function of temperature. [Figure 17] 1 shows XRPD patterns of Type IV crystals of Examples 5a and 5b. [Figure 18] 1 is a TGA / DSC pattern of the IV type crystal of Example 5a. [Figure 19] 1 is a 1H NMR spectrum of the IV type crystal of Example 5a. [Figure 20] 1 is an XRPD pattern of the V-type crystals of Example 6a. [Figure 21] 1 shows the TGA / DSC pattern of the V-type crystals of Example 6a. [Figure 22] 1 is a 1H NMR spectrum of the V-type crystals of Example 6a. [Figure 23] 1 is an XRPD pattern of the VI type crystal of Example 7a. [Figure 24] 1 is a DSC / TGA pattern of the VI type crystal of Example 7a. [Figure 25] 1 shows XRPD patterns of Type IV crystals of Example 5a before and after heating. [Figure 26] 1 shows XRPD overlay patterns of the type II crystal of Example 3a before and after heating. [Figure 27] 1 shows XRPD patterns of the type II crystal of Example 3b before and after heating. [Figure 28] 1 shows XRPD patterns of type V crystals of Example 6a before and after heating. [Figure 29] 1 is an XRPD overlay pattern of the suspension competition between Form I crystals and Form IIIA crystals in EtOAc. [Figure 30] 1 is an XRPD overlay pattern of suspension competition between Form I crystals and Form IIIA crystals in MIBK. [Figure 31] XRPD superposition pattern (I / II) of the suspension competition between Form I crystals and Form IIIA crystals in acetone / H2O. [Figure 32] XRPD superposition pattern (II / II) of the suspension competition between Form I crystals and Form IIIA crystals in acetone / H2O. [Figure 33] This is the DVS pattern of type I crystals. [Figure 34] 1 shows XRPD overlay patterns of type I crystal before and after DVS test. [Figure 35] 1 shows XRPD overlay patterns of an amorphous sample before and after DVS testing. [Figure 36] This is a DVS pattern in a moisture absorption experiment of type IIIA crystal. [Figure 37] 1 shows XRPD overlay patterns of type IIIA crystal before and after DVS test. [Figure 38] 1 shows XRPD overlay patterns of the type I crystal of Example 2a before and after evaluation of the solid stability. [Figure 39] 1 shows XRPD overlay patterns before and after evaluation of amorphous solid stability. [Figure 40] 1 shows XRPD overlay patterns of the IIIA crystal before and after evaluation of its solid-state stability. [Figure 41] 1 is a solubility diagram of the amorphous material at different pH values ​​(feed concentration 0.25 mg / mL). [Figure 42] 1 is a solubility diagram of Form I crystals at different pH values ​​(feed concentration 0.25 mg / mL). [Figure 43] 1 is a solubility diagram of amorphous and crystalline Form I at different pH values ​​(feed concentration 0.05 mg / mL). [Figure 44]This is an XRPD overlay pattern of type I crystal before and after crushing. [Figure 45] 1 shows XRPD overlay patterns of type I crystals before and after tableting. [Figure 46] 1 shows XRPD overlay patterns of type IIIA crystal before and after grinding. [Figure 47] 1 shows XRPD overlay patterns of type IIIA crystals before and after tableting. [Figure 48] 1 shows XRPD overlay patterns before and after grinding of the amorphous material. [Figure 49] 1 is an XRPD overlay pattern of amorphous before and after tableting. [Figure 50] 1 shows XRPD superimposed patterns for evaluating the stability of type I crystals against light irradiation. [Figure 51] 1 shows XRPD superimposed patterns for evaluating the light irradiation stability of the IIIA type crystal. [Figure 52] 1 shows XRPD overlay patterns for evaluating the light irradiation stability of amorphous samples. [Figure 53] 1 shows XRPD superimposed patterns for evaluating the light irradiation stability of type V crystals. [Figure 54] FIG. 1 is an HPLC diagram for evaluating the stability of type I crystals against light irradiation. [Figure 55] FIG. 1 is an HPLC diagram for evaluating the stability of type IIIA crystals under light irradiation. [Figure 56] HPLC diagram for evaluating the stability of amorphous samples against light irradiation. [Figure 57] FIG. 1 is an HPLC diagram for evaluating the stability of V-type crystals against light irradiation. DETAILED DESCRIPTION OF THE INVENTION

[0059] Example The following examples are merely illustrative of the present invention and are not intended to limit the invention in any manner.

[0060] Solvent name correspondence table [Table 1]

[0061] In the present invention, the following instruments and methods are used: Apparatus and method X-ray powder diffraction (XRPD): XRPD results are reported in X'Pert 3 and collected on an Empyrean X-ray powder diffraction analyzer, and the scanning parameters are shown in Table 1. [Table 2]

[0062] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC): TGA and DSC were collected on a TA Discovery 5500 thermogravimetric analyzer and a TA Discovery 2500 differential scanning calorimetry, respectively, and Table 2 shows the TGA and DSC test parameters. [Table 3]

[0063] Hydrogen Spectra Liquid Nuclear Magnetic (1H Solution NMR): Hydrogen Spectra Liquid nuclear magnetic spectra were collected on a Bruker 400M nuclear magnetic resonance instrument using DMSO-d6 as the solvent.

[0064] Dynamic moisture sorption (DVS): Dynamic moisture sorption (DVS) curves were collected with a DVS Intrinsic from SMS (Surface Measurement Systems). Relative humidity at 25°C was calibrated with the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS test parameters are shown in Table 3. [Table 4]

[0065] High-performance liquid chromatography (UPLC): The purity and solubility during the experiment were tested on an Agilent 1290 ultra-high performance liquid chromatograph, and the analytical conditions were shown in Tables 4 and 5. [Table 5] [Table 6]

[0066] For instruments and methods not explicitly mentioned in the "instruments and methods" section, instruments and methods known in the art can be used.

[0067] Example 1a (amorphous compound 1, repeating Example 1 of Chinese Patent Application No. 201210190520.4): According to the method of Example 1 of Chinese Patent Application No. 201210190520.4, i.e., using dichloromethane / methanol as an eluent, the crude product obtained in step 13 of Example 1 was separated and purified by silica gel column chromatography, and then rotary evaporated to obtain a white solid, which was amorphous according to XRPD analysis, and its XRPD pattern is shown in Figure 1, and its dynamic moisture sorption (DVS) is shown in Figure 2.

[0068] Example 2a: 4 mL of absolute ethanol was added to approximately 21 mg of the amorphous sample of Example 1a, and the mixture was clarified by heating and stirring in an oil bath at 85°C for 5 minutes. The resulting clarified solution was then cooled to room temperature (25°C) with stirring within 2.5 hours, and continued to be stirred at room temperature for 24 hours. The resulting solid sample was filtered and subjected to XRPD measurement.

[0069] The XRPD results are shown in Figure 3. As shown in the TGA / DSC results (Figure 4), when the sample was heated to 150°C, the weight loss was 0.77% and one sharp endothermic peak appeared at 221.3°C (onset temperature). 1 H NMR data was collected using DMSO-d6 as the solvent, and the results are shown in Figure 5. No obvious residual solvent was detected. The characterization of the type I crystals showed that they exhibited a small and moderate weight loss (less than the theoretical water content of a hemihydrate, 1.80%) and a single melting endothermic signal, suggesting that the type I crystals are an anhydrate or anhydrous crystalline form. The XRPD diffraction peak data for the type I crystals are shown in Table 6. [Table 7]

[0070] Example 2b: Approximately 20 mg of each amorphous sample was weighed out and placed in a 20-mL vial. 0.4 to 1.0 mL of a good solvent (see Table 7) was added and dissolved. The solution was filtered (through a 0.45 μm pore size PTFE filter membrane) to obtain a clear solution. While stirring the clear solution, a poor solvent (see Table 7) was added until a solid precipitated. The precipitated solid was separated by centrifugation and subjected to XRPD testing. The results are shown in Table 7. The solid obtained by adding the poor solvent was confirmed to be Type I crystals by XRPD. [Table 8] [1] After adding the poor solvent, the mixture was clarified, transferred to 5°C, and stirred to precipitate a solid. [2] : After addition of antisolvent, the mixture became clear, and after stirring at 5°C, the mixture remained clear, and after stirring at -20°C, the mixture precipitated a solid. [3]: It clarified after the addition of the antisolvent, remained clarified after stirring at 5°C and -20°C, and was then transferred to room temperature to evaporate. [4] After adding the anti-solvent, the mixture became oily, and after circulating the suspension and stirring at a temperature of 5 to 50°C, the mixture remained oily, and was then transferred to room temperature to evaporate.

[0071] Example 2c: Approximately 20 mg of each amorphous sample from Example 1a was weighed and placed in a 5-mL or HPLC vial. 0.7 to 4.0 mL of the solvent listed in Table 8 was added, and the mixture was stirred at 50°C for approximately 2 hours. The mixture was then filtered (using a PTFE membrane with a pore size of 0.45 μm) to collect the filtrate. The resulting filtrate was placed in a biochemical incubator and cooled to 5°C at a rate of 0.1°C / min. The precipitated solid was collected and subjected to XRPD analysis. The test results are shown in Table 8. A solid was obtained by slow cooling, and XRPD confirmed that it was Type I crystal. [Table 9] [1]After gradually lowering the temperature to 5°C, the mixture was clarified, transferred to -20°C and allowed to stand, yielding a solid. [2] After gradually lowering the temperature to 5°C, the mixture became clear, and was then transferred to -20°C and allowed to stand. However, no solid precipitated, and the mixture was then transferred to room temperature and allowed to evaporate.

[0072] Example 3 Type II crystal Example 3a: 71.6 mg of the Form I crystal sample of Example 2a was weighed and placed in a 3 mL vial, and 1.5 mL of ACN was added and magnetically stirred at room temperature for about 1 day. The solid was separated by centrifugation and left to dry under ambient conditions (~21 °C / 45% RH) for about 4 hours to obtain the final product, which was subjected to XRPD and TGA / DSC tests.

[0073] The XRPD results are shown in Figure 6. The TGA results are shown in Figure 7. TGA showed that a gradual weight loss of 5.90% occurred when the sample was heated to 120°C, and the DSC results showed that there were two endothermic peaks at 94.1°C and 220.9°C (onset temperature) and one exothermic peak at 191.6°C (peak value temperature). 1 The H NMR results are shown in Figure 8, and the molar ratio of ACN to API in the sample was 0.8 (5.9 wt%, consistent with the weight loss measured by TGA). Based on the characterization and heating experiment results, it was suggested that Form II crystals were ACN solvates, and that desolvation and crystal transformation occurred after heating. The XRPD diffraction peak data for Form II crystals are shown in Table 9. [Table 10]

[0074] Example 3b: 20.4 mg of the Form I crystal sample of Example 2a was weighed and placed in a 3 mL vial. 2 mL of ACN / acetone (1:1, v / v) was added. After ultrasonic treatment, the solution was filtered (0.45 μm PTFE filter membrane) to obtain a clear solution. After sealing with a sealing membrane, four small holes were drilled and the solution was placed in a fume hood to allow for slow evaporation for 5 days. When solid precipitation was observed, the solution was left under ambient conditions overnight, then transferred to room temperature and vacuum dried for approximately 1 day. The ACN / acetone (1:1, v / v) solution of the Form I crystal of Example 2a was slowly evaporated, and after solid precipitation, the solution was transferred to room temperature and vacuum dried to obtain the final product, which was then subjected to XRPD testing. The XRPD results are shown in Figure 6.

[0075] Example 3c: Approximately 19.7 mg of the amorphous sample obtained in Example 1a was weighed and placed in an HPLC vial, and 0.5 mL of ACN / HO (19:1) solvent was added to each vial. The resulting suspension was placed at 5°C and magnetically stirred (~1000 rpm) for approximately 6 days. The solid was then separated by centrifugation and subjected to XRPD testing. The XRPD result indicated that the crystal was Type II.

[0076] Example 3d: Approximately 19.9 mg of the amorphous sample obtained in Example 1a was weighed and placed in an HPLC vial, and 0.5 mL of ACN / acetone (1:1) solvent was added. The resulting suspension was magnetically stirred (about 1000 rpm) at 5°C for about 6 days, and then the solid was separated by centrifugation and subjected to XRPD testing. The XRPD result was Form II crystals.

[0077] Example 4 (Type IIIA crystals and Type IIIB crystals) Example 4a: The Type II crystal sample of Example 3a was heated to 120°C in a DSC, incubated for 5 minutes, and then cooled to room temperature to obtain Type IIIA crystal. The XRPD results are shown in Figure 9. The TGA / DSC results (Figure 10) showed that when the sample was heated to 200°C, the weight loss was 0.26%, which is lower than the theoretical water content of hemihydrate (1.80%). The DSC results showed one exothermic signal at 200.3°C (peak temperature) and one sharp endothermic signal at 219.6°C (onset temperature). 1The H NMR results (FIG. 11) showed that no obvious solvent residue was detected. Crystalline form IIIA was heated to 205°C, cooled to room temperature, and exposed to environmental conditions before and after conversion to crystalline form I. The XRPD results before and after heating are shown in FIG. 12. Characterization of crystalline form IIIA suggested that crystalline form IIIA was an anhydrate, and the exothermic signal at 198.8°C was the thermal signal of conversion to crystalline form I. The XRPD diffraction peak data for the crystalline form IIIA was shown in Table 10. [Table 11]

[0078] Example 4b: 100.0 mg of the Form I crystal sample of Example 2a was weighed and placed in a 20 mL vial. 10 mL of THF was added to dissolve the sample, and the solution was filtered (0.45 μm PTFE filter membrane) to obtain a clear solution. The filtrate was rotary evaporated at 50°C and collected to obtain the final product. The XRPD results are shown in Figure 13. The TGA / DSC results (Figure 14) showed that the sample had a weight loss of 3.49% when heated to 200°C. The DSC results showed a faint endothermic signal and an exothermic signal at 121.3°C and 144.6°C (peak temperature), respectively, and a relatively strong endothermic signal at 218.3°C (onset temperature). 1 The H NMR results (FIG. 15) showed that the molar ratio of the residual solvent THF to the API was 0.09 (1.3 wt %), which was presumed to be the solvent adsorbed on the surface. The thermal signals at 121.3°C and 144.6°C of the Form IIIB crystals are the thermal signals of the conversion to the Form I crystals. The XRPD diffraction peak data of the Form IIIB crystals was referenced in Table 11. [Table 12]

[0079] Example 5 (Type IV crystal) Example 5a: Approximately 20 mg of the amorphous sample obtained in Example 1a was weighed and placed in an HPLC vial. 0.5 mL of 1,4-dioxane solvent was added. The resulting suspension was magnetically stirred (∼1000 rpm) at room temperature for approximately 6 days. The solid was then separated by centrifugation and vacuum dried overnight at room temperature to obtain the final product, which was subjected to XRPD and TGA / DSC tests. The XRPD and TGA / DSC results are shown in Figures 17 and 18. The TGA results indicated that a gradual weight loss of 16.80% occurred when the sample was heated to 120°C. The DSC results indicated two endothermic peaks at 95.6°C and 220.5°C (onset temperature). 1 The H NMR results are shown in Figure 19, and the molar ratio of 1,4-dioxane to API in the sample was 1.2 (17.8 wt%, consistent with the weight loss observed in TGA). Based on the above characterization results and the XRPD results over temperature, the IV crystals were presumed to be 1,4-dioxane solvates. The XRPD diffraction peak data for the IV crystals were shown in Table 12. [Table 13]

[0080] Example 5b: The Type I crystal sample (500 mg) of Example 2a was stirred in 1,4-dioxane (12.5 mL) at room temperature for 2 days, and the resulting solid was dried under vacuum at room temperature for 1 day to obtain the XRPD result shown in Figure 17.

[0081] Example 6 (V-type crystal) Example 6a: 20.8 mg of the Form I crystal sample from Example 2a was weighed and placed in a 5 mL vial, followed by the addition of 4 mL of EtOH. The sample was stirred at 70°C, clarified, and cooled to 5°C within 650 minutes (at a rate of 0.1°C / min) and allowed to stand at 5°C. After removing the solution, the solid was left open to dry under ambient conditions (temperature: ∼21°C, humidity: ∼36% RH). The XRPD results are shown in Figure 20. The TGA / DSC results (Figure 21) indicated that heating the sample to 120°C resulted in a gradual weight loss of 7.64%, with two endothermic peaks at 103.1°C and 223.3°C (onset temperature).1 The H NMR results are shown in Figure 22, and the molar ratio of EtOH to API in the sample was 0.7 (6.2 wt%, which was relatively consistent with the weight loss from TGA). According to the above characterization results and the XRPD results of temperature change, it was inferred that the type V crystals were an EtOH solvate, which was desolvated after heating and transformed into the anhydrous type I crystals. The XRPD diffraction peak data for the type V crystals were referenced in Table 13. [Table 14]

[0082] Example 7 (VI type crystal) The amorphous sample (1 g) of Example 1a was dissolved in DMF (3 mL). After complete dissolution, IPA (6 mL) was added and the mixture was stirred at room temperature overnight to precipitate a solid. The solid was filtered, the filter cake was washed with isopropyl alcohol, and dried under vacuum to obtain a crystalline form of the DMF solvate, which is Type VI crystals.

[0083] The XRPD results of the VI crystal are shown in FIG. 23, the DSC results show two endothermic peaks at 147.39°C and 208.56°C, and the TGA results in FIG. 24 show that the weight loss was 0.67% when heated to 180°C.

[0084] Example 8 (Crystalline Transition) Example 8a: The Form IV crystals of Example 5a were converted to Form I crystals after heating to 120° C., cooling to room temperature and exposure to ambient conditions, and the XRPD results are shown in FIG.

[0085] Example 8b: The type II crystals of Example 3a were heated to 120°C and 210°C, respectively, cooled to room temperature, and exposed to ambient conditions. The XRPD results are shown in Figure 26. When heated to 120°C, they transformed into type IIIB crystals, and when heated to 210°C, they transformed into type I crystals.

[0086] Example 8c: The Form II crystals of Example 3b were heated to 120°C, cooled to room temperature, and exposed to ambient conditions before converting to Form I crystals, with the XRPD results shown in Figure 27. Similarly, after heating the sample, it was expected to first convert to Form IIIB crystals, but convert relatively quickly to Form I crystals under ambient conditions, and upon XRPD testing showed that it converted to Form I crystals.

[0087] Example 8d: The type IIIB crystal of Example 4a was identified by temperature-dependent XRPD, and the results are shown in Figure 16. After purging with N2 at 30°C for 20 minutes, the crystal form did not change. When heated to 120°C under N2 protection, diffraction peaks of type I crystal were observed. When heated to 170°C, mainly type I crystals were observed, with slight diffraction peaks of type IIIB crystals. When cooled to 30°C under N2 protection, no further transformation occurred.

[0088] Example 8e: The type V crystals of Example 6a were converted to type I crystals after heating to 120° C., cooling to room temperature and exposure to ambient conditions, and the XRPD results were shown in FIG.

[0089] Example 8f (Suspension competition experiment between type I crystals and type IIIA crystals) The examples relate to suspension competition experiments between Form I crystals and Form IIIA crystals in EtOAc and MIBK at room temperature and 50° C., and in acetone / H 2 O with different water activities at room temperature.

[0090] Approximately 15 mg of the Type I crystal sample of Example 2a was weighed and placed in an HPLC bottle, and 1 mL of the corresponding solvent was added. The mixture was stirred at the corresponding temperature for 4 hours or overnight. The mixture was filtered (using a 0.45 μm PTFE filter membrane) to obtain a saturated solution. Equal masses (approximately 5 mg each) of the Type I crystal sample of Example 2 and the Type IIIA crystal sample of Example 4a were weighed into new HPLC bottles, and the saturated solution obtained in Step 1 was added. The mixture was stirred at the corresponding temperature, and the solid wet sample was subjected to XRPD testing (covered with a Kapton film to avoid potential crystal transitions caused by solvent evaporation during the testing process).

[0091] The results of the suspension competition are summarized in Table 14. The XRPD results are shown in Figures 29, 30, 31 and 32. [Table 15] where a w :Theoretical water activity.

[0092] The XRPD comparison results showed that under all conditions, physical mixtures of Form I crystals and Form IIIA crystals were converted to Form I crystals after suspension competition, and that the anhydrous Form I crystals were thermodynamically more stable than Form IIIA crystals at room temperature to 50°C and under anhydrous conditions, as well as at room temperature and water activity of 0 to 1.

[0093] Example 9 (Hygroscopicity of Form I Crystals) The hygroscopicity of the Form I crystal of Example 2a, the amorphous material of Example 1a, and the Form IIIA crystal of Example 4a was evaluated by a dynamic moisture sorption (DVS) test at 25°C.

[0094] The DVS results for the type I crystal are shown in Figure 33. The weight gain upon water absorption at 25°C / 80%RH was 0.047%, indicating almost no hygroscopicity. The XRPD comparison results (Figure 34) indicated that the type I crystal did not undergo crystal form transformation after the DVS test.

[0095] The DVS results for the amorphous sample are shown in Figure 2. As the humidity increased from 50% RH to 95% RH, the weight gain during water absorption decreased continuously. This suggests that the amorphous sample underwent a crystal form transformation during the humidity increase, and that the water or organic solvent adsorbed or encapsulated in the original sample was removed under the action of N2 purging. The XRPD comparison results (Figure 35) showed that the amorphous sample transformed into type I crystal after the DVS test.

[0096] The DVS results for the IIIA crystal are shown in Figure 36. The weight gain upon water absorption at 25°C / 80%RH was 0.060%, indicating that the IIIB crystal is almost non-hygroscopic. The XRPD comparison results (Figure 37) indicated that the IIIB crystal did not undergo crystal form transformation after the DVS test.

[0097] Example 10 (Solid state stability of type I crystal) To evaluate the solid-state stability of the Form I crystals, Form IIIA crystals, and amorphous materials, appropriate amounts of the Form I crystals of Example 2a, the amorphous sample of Example 1a, and the Form IIIA crystals of Example 4a were weighed and stability experiments were conducted under the following conditions: 60°C / sealed / 1 day, 25°C / 60% RH / open / 1 week, and 40°C / 75% RH / open / 1 week. The samples under different conditions were examined for crystalline form by XRPD to evaluate their physical stability, and their purity and chemical stability were evaluated by HPLC. The evaluation results are summarized in Table 15, and the XRPD results are shown in Figures 38, 39, and 40 (a faint diffraction peak of Form I crystals was observed in the area marked with an asterisk in Figure 40). The stability results showed that the type I crystal did not undergo crystal form conversion or purity loss under any of the three evaluation conditions, indicating that the type I crystal has good physical and chemical stability under the evaluation conditions. The purity of the amorphous material did not change significantly under any of the three evaluation conditions, but all of them converted to type I crystal. The type IIIA crystal showed a faint diffraction peak of type I crystal after being stored in a sealed container at 60°C for 1 day, and did not change in crystal form after being stored in the open container for 1 week at 25°C / 60%RH and 40°C / 75%RH. [Table 16] *: All amorphous samples were converted to type I crystals after stability evaluation. #: A faint diffraction peak of type I crystal was observed.

[0098] Example 11 (Solubility) The dynamic solubility of the crystalline form I of Example 2a and the amorphous form of Example 1a was tested at room temperature and under different pH conditions (1M HCl and pH 2.0 / 4.5 / 6.8 / 7.4 buffer solution). The specific steps are as follows: (1) Approximately 2.5 mg of a Form I crystal or amorphous sample was weighed and placed in a 20 mL glass bottle, and 10 mL of buffer solutions of different pH values ​​was added. Alternatively, approximately 2.0 mg of a Form I crystal or amorphous sample was weighed and placed in a 20 mL glass bottle, and 8 mL of hydrochloric acid (1 M) was added. (2) After shaking (~500 rpm) for 3 minutes at room temperature, 0.8-1 mL of sample was drawn up with a syringe and filtered (0.45 μm PTFE filter membrane) before HPLC testing. (3) 100 μL of the solution obtained from the amorphous material was taken and diluted 10 times with the corresponding buffer solution for use. If solid precipitation occurred in the undiluted solution, the diluted sample was tested; if no solid precipitation occurred, the undiluted sample was tested. Since the solution obtained from the type I crystal was estimated to have relatively low solubility, the clear solution after filtration was directly tested by HPLC without dilution.

[0099] The solubilities of Form I crystals and amorphous form under different pH conditions are shown in Table 16. The results showed that the solubility of Form I crystals remained almost unchanged at different pH levels, while the solubility of amorphous form changed significantly with pH (Figure 41). On the other hand, the in vivo dissolution of Form I crystals was more stable (Figure 42), making it easier to obtain in vivo pharmacokinetic results with stable blood drug concentrations, which is advantageous for avoiding drug safety risks caused by excessive fluctuations in blood drug concentrations. Similar results were also observed when the supply concentration was 0.05 mg / mL (Figure 43). [Table 17] LOQ=0.28 μg / mL.

[0100] Example 12 (Powder Properties) To understand the powder flow properties of the crystalline Form I and amorphous materials, powder properties including angle of repose, bulk density, and tapped density were evaluated for the crystalline Form I of Example 2a and the amorphous sample of Example 1a.

[0101] Bulk density and tapped density: A certain mass of the sample to be evaluated was added to a 5 mL measuring cylinder and the volume was recorded. The bulk density was calculated by dividing the mass of the sample by the volume. The measuring cylinder was tapped 200 times, the final volume was recorded, and the tapped density was calculated by dividing the mass of the sample by the final volume. Each parameter was tested three times in parallel.

[0102] Angle of repose: The funnel was fixed perpendicular to the base, and the material was slowly added into the funnel. The base formed a uniform vertebra of material. The height of the vertebra and the diameter of the base were measured. Three parallel measurements were taken.

[0103] The bulk density / tap density results are summarized in Table 17. According to the results, the average bulk density and tap density of the I-type crystals were 0.34 g / cm, respectively. 3 and 0.46 g / cm 3 The calculated Kerr index is 26%, and the average amorphous bulk density and tap density are 0.31 g / cm, respectively. 3 and 0.46 g / cm 3 and the calculated Carr index is 33%.

[0104] The results of the angle of repose are summarized in Table 18. The results showed that the angles of repose of the crystalline form I and amorphous samples were 27.7° and 26.7°, respectively. The comprehensive evaluation results showed that the crystalline form I and amorphous samples had relatively similar Kerr indices and angles of repose, and the crystalline form I and amorphous samples had similar fluidity. [Table 18] Carr index = (tap density - bulk density) / tap density. [Table 19] Formula for calculating angle of repose α: α=tan -1 (h / D), where h is the vertebral body height and D is the vertebral body diameter.

[0105] Example 13 (Mechanical Stability) The Type I crystals of Example 2a, the Type IIIA crystals of Example 4a, and the amorphous material of Example 1a were manually crushed and tableted using a press (pressure 350 MPa). XRPD tests were performed on the crushed and tableted samples to evaluate their mechanical stability. The XRPD results are shown in Figures 44, 45, 46, 47, 48, and 49. The evaluation results showed that after crushing and tableting, the Type I crystals did not undergo crystal form conversion and the crystallinity did not decrease significantly. The Type III crystals did not change crystal form after crushing, but the crystallinity decreased slightly. After tableting, the crystal form did not change, but the crystallinity decreased significantly. The amorphous material converted to Type I crystals after crushing and tableting.

[0106] Based on the characterization and identification of the crystal form, Form I crystal was selected for evaluation of hygroscopicity, solid-state stability, solubility, powder properties, and mechanical stability. The amorphous form was also evaluated for comparison with the properties of Form I crystal. The DVS results indicated that Form I crystal was nearly non-hygroscopic, and no crystal form conversion occurred after the DVS test. The amorphous form converted to Form I crystal after the DVS test. The solid-state stability results indicated that Form I crystal underwent no crystal form conversion or purity loss after being stored in a sealed container at 60°C for one day, and then left open at 25°C / 60% RH and 40°C / 75% RH for one week. Form I crystal possessed good physical and chemical stability under the evaluation conditions. The purity of the amorphous form did not change significantly under any of the three evaluation conditions, but all converted to Form I crystal. The powder property test results for Form I crystal indicated that Form I crystal and the amorphous sample had similar flow properties. According to the mechanical stability results, after tableting (350 MPa) and manual crushing (about 3 minutes), the type I crystal did not undergo crystal form transformation, and the crystallinity did not decrease significantly, and the amorphous material transformed into the type I crystal.

[0107] According to the characterization data and evaluation results, the Form I crystal did not undergo any crystal form transformation under any of the evaluation conditions, whereas the amorphous form transformed into the Form I crystal after the DVS, solid state stability, solubility, and mechanical stability tests, indicating that the Form I crystal has superior physical stability compared to the amorphous form.

[0108] Example 14 (Stability under light irradiation) The I-type crystal of Example 2a, the IIIA-type crystal of Example 4a, the V-type crystal of Example 6a, and the amorphous of Example 1a were each irradiated with light (white light 5800 to 5890 Lux + ultraviolet light 7.9 to 8.7 W / m 2 ), and some samples were taken at 6 hours and 24 hours, respectively, and used for XRPD and HPLC purity tests to evaluate stability. The XRPD results are shown in Figures 50, 51, 52, and 53. None of the Form I crystals, Form IIIA crystals, Form V crystals, and amorphous crystals underwent crystal form conversion after 24 hours under light irradiation conditions. Under the reference condition, which was protected from light, the Form I crystals, Form IIIA crystals, and Form V crystals underwent crystal form conversion after 24 hours, and a faint diffraction peak of Form I crystals was observed in the amorphous crystal after 6 hours and 24 hours. The HPLC results are shown in Figure 54, Figure 55, Figure 56, Figure 57 and Table 19. After 24 hours of light irradiation, the purity of Type I crystal under light irradiation conditions slightly decreased from 100.00 area% to 99.14 area% (content of impurity A: 0.79%), while the purity of Type IIIA crystal, Type V crystal and amorphous under light irradiation conditions decreased from 100.00 area% to 98.46 area% (content of impurity A: 1.34%), 92.05 area% (content of impurity A: 7.95%) and 90.42 area% (content of impurity A: 8.83%), respectively. The purity of the four samples under the reference conditions in the dark did not change significantly (in Figure 54, Figure 55 and Figure 56, the 6-hour light-irradiated sample and the test for photolysis impurities were performed using two HPLC test sequences, so the retention times of the impurities may be slightly different). According to the results of the light irradiation stability, the type I crystal exhibited superior light irradiation stability compared to the type IIIA crystal, the type V crystal and the amorphous material. [Table 20]

[0109] Example 15 (Pharmacokinetics experiment in SD rats) 2.39 mg of Form I crystals from Example 2a, 2.38 mg of Form IIIA crystals from Example 4a, and 2.46 mg of amorphous material from Example 1 were used as test substances. 7.967 mL, 7.906 mL, and 8.193 mL of 0.5% CMC-Na aqueous solution were added, respectively, to prepare gavage formulations with a final concentration of 0.3 mg / mL. Three male SD rats were gavaged with the three test substance solutions at a dose of 3 mg / kg body weight. 0.15 mL of blood was collected from the jugular sinus at time points 0, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 10 h, and 24 h. The collected whole blood was placed in an EDTA-K2 anticoagulant tube, thoroughly mixed, and centrifuged (1500-1600 g) for 10 min. Plasma was separated and used for biological analysis. The concentrations of the test article in the plasma samples were measured using an LC-MS / MS analytical method (instrument model: Triple Quad 5500, chromatography column: Agilent ZORBAX XDB-C18, flow rate: 0.50 mL / min, sample input: 2 μL, mobile phase A: water [0.1% formic acid + 5 mM ammonium acetate], mobile phase B: acetonitrile [0.1% formic acid]). The corresponding pharmacokinetic parameters (Table 20) were calculated using a non-compartmental model in PharsightPhoenix 8.3. As can be seen from the data, the AUC 0-t were 9820ng·h / mL and 8890ng·h / mL, respectively, and C max were 1160 ng / mL and 1030 ng / mL, respectively, indicating sufficient plasma exposure of the drug in vivo, favorable pharmacokinetic properties, and that the type I crystal has higher exposure and blood drug concentration than the type IIIA crystal, potentially allowing for a lower dosage. [Table 21]

[0110] Example 16: Inhibition of proliferation of NRAS mutant melanoma cell lines All cells were provided by Beijing Cancer Hospital. Human melanoma cell line SK-MEL-2 (NRAS Q61R) was cultured in MEM + 10% FBS + 1% penicillin and streptomycin at 37°C in an incubator with 5% carbon dioxide. Human melanoma cell line HMVII (NRAS Q61K) was cultured in F12K + 10% FBS + 1% penicillin and streptomycin at 37°C in an incubator with 5% carbon dioxide.

[0111] When the cultured cells reached 80% or greater confluence, the adherent cells were digested with trypsin, centrifuged to collect and count the cell pellet, and 90 μL of the cell suspension was seeded into a 96-well plate at the appropriate density. After 24 hours, a gradient dilution series of compound (form I crystals) (concentrations ranging from 0.15 nM to 10 μM, diluted 4-fold) was added in a volume of 10 μL per well. Three parallel wells were used for each concentration, for a total of nine concentration points. An equal volume of 5% DMSO was added as a control, with the final DMSO concentration at 0.5%. Three days after drug treatment, cell activity was detected using MTT. 10 μL of MTT was added to each well and the wells were incubated in an incubator for 4 hours. The supernatant was then discarded, and 150 μL of DMSO was added to dissolve the formazan crystals. The absorbance at 490 nM was detected using a plate reader. The above experiment was repeated three times, and the dose-effect curve was constructed using GraphPad Prism8 software, and IC50 was calculated. The results are shown as the mean ± SD value of IC50 from three experiments (Tables 21 and 22). [Table 22] [Table 23]

[0112] Example 17: Inhibition of RAS or RAF mutants The tetrazolium salt (MTS) method was used to detect the in vitro antiproliferative activity of compound 1 against RAS-mutated or RAF-mutated tumor cell lines and RAS / RAF wild-type tumor cell lines, as well as normal human cell lines.

[0113] Adherent cells in the logarithmic growth phase were digested with trypsin or centrifuged to collect and count suspended cells. 150 μL of cells were seeded into a 96-well plate. After 24 hours, 50 μL of compound (form I crystals) diluted in medium at 4x the final concentration was added per well (concentration range: 0.15 nM to 1000 nM, 3-fold gradient dilution). Control wells were treated with 2% DMSO at an equal volume, with a final DMSO concentration of 0.5%. After 72 hours of incubation, cell activity was detected using MTS. The specific method was as follows: for adherent cells, the medium was discarded, and 20 μL of MTS and 100 μL of cell culture medium were added to each well. For suspended cells, 20 μL of MTS was added directly. After 1 to 4 hours of incubation in an incubator, OD490 was measured using OD650 as a reference. Dose-effect curves were generated using GraphPad Prism software, and IC 50 was calculated, and the results are shown in Table 23. [Table 24]

[0114] Example 18: A multicenter, single-arm, phase II study evaluated the efficacy and safety of Form I crystalline Compound 1 in patients with NRAS-mutant advanced melanoma. Patients received 12 mg of the product orally twice daily until intolerable toxicity, disease progression (as assessed by investigators using RECIST 1.1), consent to withdrawal, death, or investigator-determined risk outweighed benefit. A total of 100 subjects had been enrolled through February 19, 2023, with 95.0% (95 / 100) of subjects included in the full analysis set (FAS).

[0115] Main treatment effect measures: In the FAS population, the ORR (objective response rate) assessed by an independent radiological review committee (IRRC) was 35.8% (34 / 95 cases) (95% CI: 26.2%, 46.3%).

[0116] In the FAS population, according to the IRRC evaluation results, the median PFS (progression-free survival) was 4.2 months (95% CI: 3.5, 5.6), the DCR was 72.6% (69 / 95 cases) (95% CI: 62.5%, 81.3%), and the median DoR was 6.1 months (95% CI: 3.9, 8.9).

[0117] Compound 1 demonstrated favorable antitumor therapeutic effects in patients with NRAS-mutated advanced melanoma, with an ORR of 35.8% as assessed by IRRC, demonstrating significantly better efficacy than clinical studies of similar drugs.

Claims

1. A crystalline polymorph of formula (I), 【Chemistry 1】 Here, n is 0 or 1, and X is acetonitrile, water, 1,4-dioxane, ethanol, methanol, dimethylformamide, acetone, or a mixture thereof.

2. 2. The crystalline polymorph of claim 1, wherein n is 0.

3. 2. The crystalline polymorph of claim 1, wherein n is 1 and X is acetonitrile, water, 1,4-dioxane, or ethanol.

4. 2. The crystalline polymorph according to claim 1, wherein n is 0, the crystalline polymorph is a type I crystal, and the X-ray powder diffraction pattern of the type I crystal has characteristic diffraction peaks at 2θ positions of 16.71°±0.2°, 21.82°±0.2°, and 23.75°±0.2°, using Cu-Kα radiation.

5. 5. The crystalline polymorph of claim 4, wherein the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at 2θ positions of 7.48°±0.2° and 22.36°±0.2°.

6. 6. The crystalline polymorph of claim 5, wherein the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at the 2θ positions of 5.3°±0.2°, 24.57°±0.2°, and 27.08°±0.2°.

7. 7. The crystalline polymorph of claim 6, wherein the X-ray powder diffraction pattern of the Form I crystal also has characteristic diffraction peaks at the following 2θ positions: 11.81°±0.2°, 15.83°±0.2°, 17.92°±0.2°, 18.95°±0.2°, and 19.17°±0.2°.

8. The X-ray powder diffraction pattern of the Form I crystal was 5.30°±0.2°, 7.48°±0.2°, 11.81°±0.2°, 14.85°±0.2°, 15.83°±0.2°, 16.71°±0.2°, 17.92°±0.2°, 18.95°±0.2°, 19.17°±0.2°, 19.43°±0.2°, 21.14°±0.2°, 21.82°±0.2°, 22.36°±0.2°, 23.

5. The crystalline polymorph of claim 4, having characteristic diffraction peaks at 2θ positions of 75°±0.2°, 24.57°±0.2°, 27.08°±0.2°, 27.83°±0.2°, 28.88°±0.2°, 31.20°±0.2°, 31.92°±0.2°, 32.40°±0.2°, 33.91°±0.2°, 35.83°±0.2°, 37.51°±0.2°, and 39.04°±0.2°.

9. 5. The crystalline polymorph of claim 4, wherein the X-ray powder diffraction pattern of the Form I crystal is essentially as shown in FIG.

10. The crystalline polymorph of claim 4, wherein the Form I crystal has the TGA pattern and / or DSC pattern shown in Figure 4.

11. 2. The crystalline polymorph according to claim 1, wherein n is 1, X is acetonitrile, the crystalline polymorph is a Type II crystal, and the X-ray powder diffraction pattern of the Type II crystal has characteristic diffraction peaks at 2θ positions of 6.35°±0.2°, 20.34°±0.2°, 22.41°±0.2°, 22.6°±0.2°, 24.99°±0.2°, 26.05°±0.2°, and 28.71°±0.2°, using Cu-Kα radiation.

12. 12. The crystalline polymorph of claim 11, wherein the X-ray powder diffraction pattern of the Form II crystal also has characteristic diffraction peaks at the following 2θ positions: 9.18°±0.2°, 16.03°±0.2°, 18.25°±0.2°, 27.07°±0.2°, 29.08°±0.2°, and 33.93°±0.2°.

13. 13. The crystalline polymorph of claim 12, wherein the X-ray powder diffraction pattern of the Form II crystal also has characteristic diffraction peaks at the following 2θ positions: 14.44°±0.2°, 24.64°±0.2°, 26.41°±0.2°, 32.27°±0.2°, 32.68°±0.2°, 37.07°±0.2°, and 39.51°±0.2°.

14. The X-ray powder diffraction pattern of the Form II crystal is 6.35°±0.2°, 9.18°±0.2°, 9.91°±0.2°, 14.44°±0.2°, 16.03°±0.2°, 18.25°±0.2°, 19.77°±0.2°, 20.34°±0.2°, 21.81°±0.2°, 22.41°±0.2°, 22.60°±0.2°, 23.84°±0.2°, 24.64°±0.2°, 24.99°±0.2°, 25.43°±0.2°, 26.05°±0.2°, 26.41°±0.2° 12. The crystalline polymorph of claim 11, having characteristic diffraction peaks at the following 2θ positions: 27.07°±0.2°, 28.71°±0.2°, 29.08°±0.2°, 29.81°±0.2°, 31.16°±0.2°, 31.57°±0.2°, 32.27°±0.2°, 32.68°±0.2°, 33.93°±0.2°, 34.19°±0.2°, 35.42°±0.2°, 37.07°±0.2°, 37.56°±0.2°, 38.69°±0.2°, and 39.51°±0.2°.

15. 12. The crystalline polymorph of claim 11, wherein the X-ray powder diffraction pattern of said Form II crystal is essentially as shown in Figure 6.

16. 12. The crystalline polymorph of claim 11, wherein the Form II crystal has the TGA pattern and / or DSC pattern shown in Figure 7.

17. 2. The crystalline polymorph according to claim 1, wherein n is 0, the crystalline polymorph is a Type IIIA crystal, and the X-ray powder diffraction pattern of the Type IIIA crystal has characteristic diffraction peaks at 2θ positions of 6.59°±0.2°, 22.69°±0.2°, 20.32°±0.2°, 23.62°±0.2°, 23.91°±0.2°, and 24.15°±0.2°, using Cu-Kα radiation.

18. 18. The crystalline polymorph of claim 17, wherein the X-ray powder diffraction pattern of Form IIIA also has characteristic diffraction peaks at the following 2θ positions: 10.8°±0.2°, 17.14°±0.2°, 13.75°±0.2°, 21.59°±0.2°, and 26.01°±0.2°.

19. 20. The crystalline polymorph of claim 18, wherein the X-ray powder diffraction pattern of Form IIIA also has characteristic diffraction peaks at the following 2θ positions: 18.71°±0.2°, 21.97°±0.2°, 25.54°±0.2°, 27.13°±0.2°, 27.59°±0.2°, and 30.51°±0.2°.

20. The X-ray powder diffraction pattern of the Form IIIA crystal is 6.59°±0.2°, 9.9°±0.2°, 10.8°±0.2°, 13.09°±0.2°, 13.75°±0.2°, 17.14°±0.2°, 17.87°±0.2°, 18.71°±0.2°, 19.19°±0.2°, 20.32°±0.2°, 21.59°±0.2°, 21.97°±0.2°, 22.69°±0.2°, 23.62°±0.2°, 23.91°±0.2°, 24.15°±0.2°.

18. The crystalline polymorph of claim 17, having characteristic diffraction peaks at the following 2θ positions: 25.54°±0.2°, 26.01°±0.2°, 27.13°±0.2°, 27.59°±0.2°, 28.83°±0.2°, 29.24°±0.2°, 30.51°±0.2°, 31.13°±0.2°, 31.79°±0.2°, 33.6°±0.2°, 34.14°±0.2°, 36.08°±0.2°, 36.67°±0.2°, and 37.26°±0.2°.

21. 18. The crystalline polymorph of claim 17, wherein the X-ray powder diffraction pattern of Form IIIA is essentially as shown in Figure 9.

22. 18. The crystalline polymorph of claim 17, wherein the Form IIIA crystal has the TGA and / or DSC pattern shown in Figure 10.

23. 2. The crystalline polymorph according to claim 1, wherein n is 0, the crystalline polymorph is a IIIB type crystal, and the X-ray powder diffraction pattern of the IIIB type crystal has characteristic diffraction peaks at 2θ positions of 6.53°±0.2°, 13.69°±0.2°, 18.6°±0.2°, 20.19°±0.2°, 21.52°±0.2°, and 22.64°±0.2°, using Cu-Kα radiation.

24. 24. The crystalline polymorph of claim 23, wherein the X-ray powder diffraction pattern of the Form IIIB crystal also has characteristic diffraction peaks at the following 2θ positions: 10.75°±0.2°, 17.07°±0.2°, 21.93°±0.2°, 26.13°±0.2°, 23.57°±0.2°, and 30.46°±0.2°.

25. 25. The crystalline polymorph of claim 24, wherein the X-ray powder diffraction pattern of the Form IIIB crystal also has characteristic diffraction peaks at the following 2θ positions: 13.05°±0.2°, 16.63°±0.2°, 20.82°±0.2°, 24.01°±0.2°, 27.55°±0.2°, and 31.79°±0.2°.

26. The X-ray powder diffraction pattern of the Type IIIB crystal is 6.53°±0.2°, 10.75°±0.2°, 12.62°±0.2°, 13.05°±0.2°, 13.69°±0.2°, 16.63°±0.2°, 17.07°±0.2°, 18.60°±0.2°, 19.59°±0.2°, 20.19°±0.2°, 20.82°±0.2°, 21.52°±0.2°, 21.93°±0.2°, 22.64°±0.2°.

24. The crystalline polymorph of claim 23, having characteristic diffraction peaks at the following 2θ positions: 23.57°±0.2°, 24.01°±0.2°, 25.46°±0.2°, 26.13°±0.2°, 27.55°±0.2°, 30.46°±0.2°, 31.04°±0.2°, 31.79°±0.2°, 32.81°±0.2°, 33.54°±0.2°, 34.06°±0.2°, and 34.46°±0.2°.

27. 24. The crystalline polymorph of claim 23, wherein the X-ray powder diffraction pattern of said Form IIIB crystal is essentially as shown in Figure 13.

28. 24. The crystalline polymorph of claim 23, wherein the Form IIIB crystal has the TGA and / or DSC pattern shown in Figure 14.

29. 2. The crystalline polymorph according to claim 1, wherein n is 1, X is 1,4-dioxane, the crystalline polymorph is a Type IV crystal, and the X-ray powder diffraction pattern of the Type IV crystal has characteristic diffraction peaks at 2θ positions of 8.56°±0.2°, 13.29°±0.2°, 17.69°±0.2°, 19.75°±0.2°, and 22.45°±0.2°, using Cu-Kα radiation.

30. 30. The crystalline polymorph of claim 29, wherein the X-ray powder diffraction pattern of the Form IV crystal also has characteristic diffraction peaks at the following 2θ positions: 5.26°±0.2°, 18.29°±0.2°, 31.83°±0.2°, 25.68°±0.2°, 22.86°±0.2°, 32.81°±0.2°, and 23.44°±0.2°.

31. 31. The crystalline polymorph of claim 30, wherein the X-ray powder diffraction pattern of the Form IV crystal also has characteristic diffraction peaks at the following 2θ positions: 126.57°±0.2°, 27.52°±0.2°, 35.69°±0.2°, 21.09°±0.2°, 20.35°±0.2°, and 31.43°±0.2°.

32. The X-ray powder diffraction pattern of the Type IV crystal is 5.26°±0.2°, 8.56°±0.2°, 9.85°±0.2°, 13.29°±0.2°, 17.69°±0.2°, 18.29°±0.2°, 19.75°±0.2°, 20.35°±0.2°, 21.09°±0.2°, 22.45°±0.2°, 22.86°±0.2°, 23.44°±0.2°, 24.44°±0.2°.

30. The crystalline polymorph of claim 29, having characteristic diffraction peaks at the following 2θ positions: 25.68°±0.2°, 26.57°±0.2°, 27.52°±0.2°, 28.40°±0.2°, 29.78°±0.2°, 31.43°±0.2°, 31.83°±0.2°, 32.81°±0.2°, 34.29°±0.2°, 35.69°±0.2°, and 37.72°±0.2°.

33. 30. The crystalline polymorph of claim 29, wherein the X-ray powder diffraction pattern of said Form IV crystal is essentially as shown in Figure 17.

34. 30. The crystalline polymorph of claim 29, wherein the Form IV crystal has the TGA pattern and / or DSC pattern shown in Figure 18.

35. 2. The crystalline polymorph according to claim 1, wherein n is 1, X is ethanol, the crystalline polymorph is a V-type crystal, and the X-ray powder diffraction pattern of the V-type crystal has characteristic diffraction peaks at 2θ positions of 6.21°±0.2°, 8.47°±0.2°, 15.62°±0.2°, 21.73°±0.2°, 25.53°±0.2°, 25.94°±0.2°, and 28.05°±0.2°, using Cu-Kα radiation.

36. 36. The crystalline polymorph of claim 35, wherein the X-ray powder diffraction pattern of the Form V crystals also has characteristic diffraction peaks at the following 2θ positions: 9.61°±0.2°, 17.55°±0.2°, 19.25°±0.2°, 22.22°±0.2°, 23.12°±0.2°, 32.92°±0.2°, and 34.22°±0.2°.

37. 37. The crystalline polymorph of claim 36, wherein the X-ray powder diffraction pattern of the Form V crystal also has characteristic diffraction peaks at the following 2θ positions: 9.06°±0.2°, 20.07°±0.2°, 28.49°±0.2°, 30.21°±0.2°, 31.25°±0.2°, 35.47°±0.2°, and 38.94°±0.2°.

38. The X-ray powder diffraction pattern of the Type V crystal was 6.21°±0.2°, 8.47°±0.2°, 25.94°±0.2°, 15.62°±0.2°, 25.53°±0.2°, 28.05°±0.2°, 21.73°±0.2°, 17.55°±0.2°, 32.92°±0.2°, 23.12°±0.2°, 22.22°±0.2°, 19.25°±0.2°, 34.22°±0.2°, 9.61°±0.2°, 9.06°±0.2°, 38.94°±0.2° 36. The crystalline polymorph of claim 35, having characteristic diffraction peaks at the following 2θ positions: 31.25°±0.2°, 35.47°±0.2°, 28.49°±0.2°, 30.21°±0.2°, 20.07°±0.2°, 39.78°±0.2°, 14.26°±0.2°, 24.32°±0.2°, 16.95°±0.2°, 32.33°±0.2°, 36.43°±0.2°, 24.94°±0.2°, 12.42°±0.2°, and 37.86°±0.2°.

39. 36. The crystalline polymorph of claim 35, wherein the X-ray powder diffraction pattern of said Form V crystals is essentially as shown in Figure 20.

40. 36. The crystalline polymorph of claim 35, wherein the Form V crystal has the TGA pattern and / or DSC pattern shown in Figure 21.

41. The crystalline polymorph according to any one of claims 1 to 40, wherein the crystalline polymorph is essentially free of impurity (A).

42. 41. The crystalline polymorph of claim 40, wherein the crystalline polymorph of Compound 1 contains less than 0.15% by weight of impurity (A) relative to the crystalline polymorph.

43. The crystalline polymorph of claim 4, wherein the Form I crystal is essentially free of impurity (A).

44. 44. The crystalline polymorph of claim 43, wherein the Form I crystal contains less than 0.15% by weight of impurity (A) relative to the Form I crystal.

45. 1. A pharmaceutical composition comprising:

45. A pharmaceutical composition comprising the crystalline polymorph of any one of claims 1 to 44 and a pharmaceutically acceptable carrier and / or excipient.

46. The pharmaceutical composition of claim 45, wherein the crystalline polymorph is Form I crystal.

47. 45. A method for treating tumors, chronic inflammatory diseases, inflammatory bowel diseases, skin diseases, diabetes, eye diseases, diseases associated with angiogenesis or revascularization in a mammal, diseases associated with chronic pain and other diseases regulated by the Mek cascade in a mammal, comprising administering to said mammal the crystalline polymorph of any one of claims 1 to 44.

48. 45. The crystalline polymorph of any one of claims 1 to 44 for use in treating tumors, chronic inflammatory diseases, inflammatory bowel diseases, skin diseases, diabetes, eye diseases, diseases associated with angiogenesis or revascularization in mammals, diseases associated with chronic pain and other diseases regulated by the Mek cascade in mammals.

49. 45. Use of the crystalline polymorph of any one of claims 1 to 44 in a preparation for treating tumors, chronic inflammatory diseases, inflammatory bowel diseases, skin diseases, diabetes, eye diseases, diseases associated with angiogenesis or revascularization in mammals, diseases associated with chronic pain and other diseases regulated by the Mek cascade in mammals.

50. A method for preparing the Form I crystal of claim 4, comprising: a) adding an amorphous sample of Compound 1 to a solvent, then heating to a temperature of about 70° C. or higher, allowing the resulting clear solution to cool to room temperature, keeping it at room temperature and stirring continuously, allowing the solid to precipitate, filter and dry; or b) dissolving an amorphous sample of Compound 1 in a good solvent, filtering to obtain a clear solution, and adding a poor solvent while stirring the clear solution until a solid precipitates; or c) dissolving an amorphous sample of Compound 1 in a solvent, stirring at about 50°C, then filtering to obtain a filtrate, cooling the resulting filtrate to about 5°C, and collecting the precipitated solid.

51. 51. The method of claim 50, wherein the heating temperature in method 1) is about 75°C to about 100°C, about 80°C to about 90°C, or about 75°C or about 85°C.

52. 51. The method of claim 50, wherein the room temperature in method 1) is about 20 to about 25°C.

53. 51. The method of claim 50, wherein the sustained stirring time in method 1) is from about 1 to about 12 hours, from about 1 to about 8 hours, from about 1 to about 5 hours or more, or from about 24 to about 96 hours or more.

54. 51. The method of claim 50, wherein the clarified solution in method 1) is cooled to room temperature within about 2 to about 5 hours, or within about 2.5 to about 3 hours.

55. The method according to claim 50, wherein in method 1), after continuing to incubate and stir at room temperature, the temperature can be further lowered to about 0 to about 10°C, and the mixture can be incubate and stirred.

56. 51. The method of claim 50, wherein the mixture is incubated at about 0 to about 10° C. and stirred for about 1 to about 12 hours, about 1 to about 8 hours, about 1 to about 5 hours, or more.

57. 51. The method of claim 50, wherein the solvent in method 1) is water, methanol, ethanol, isopropyl alcohol, acetone, methyl isobutyl ketone, 2-butanone, ethyl acetate, isopropyl acetate, methyl tert-butyl ether, tetrahydrofuran, anisole, 2-methyltetrahydrofuran, cyclopentyl methyl ether, 1,4-dioxane, acetonitrile, dichloromethane, toluene, meta-xylene, n-heptane, n-hexane, n-pentane, dimethyl sulfoxide, dimethylacetamide, N-methylpyrrolidone, or a mixture thereof.

58. 51. The method of claim 50, wherein the solvent in method 1) is ethanol.

59. The method of claim 50, wherein the good solvent in method 2) is a solvent in which compound 1 can be dissolved, and the poor solvent in method 2) is a solvent in which compound 1 cannot be dissolved.

60. The good solvent in method 2) is MEK, 1,4-dioxane or DMSO, and the poor solvent in method 2) is MTBE, EtOAc, CHCl3, n-heptane, Anisole, EtOAc, H ２ 51. The method of claim 50, wherein the solvent is 0, IPAc, CPME, DCM or toluene.

61. 51. The method of claim 50, wherein the cooling rate in method 3) is 0.1°C / min.

62. 51. The method of claim 50, wherein the solvent in method 3) is MIBK, methyl acetate, 2-MeTHF, or acetone / EtOH (1:1).

63. 36. A method for preparing the Type V crystals of claim 35, comprising:

1. A method comprising adding an amorphous sample of Compound 1 to a solvent, followed by heating at a temperature of about 70°C or less, and cooling the resulting clear solution to about 0 to about 10°C, allowing it to stand at that temperature until a solid precipitates, and drying.

64. 64. The method of claim 63, wherein the method comprises heating at about 70°C, about 60°C, or about 50°C.

65. 64. The method of claim 63, wherein the ramp-down rate is about 0.1 to about 0.5° C. / minute.

66. 64. The method of claim 63, wherein the ramp-down rate is about 0.1°C / minute.

67. 64. The method of claim 63, wherein the resulting clear solution is cooled to about 5°C.

68. 1. A method of treating a mammalian RAS or RAF mutant cancer, comprising: 4-fluoro-5-(2-fluoro-4-iodophenylamino)-1H-benzo[d]thiazole-6-carboxylic acid (2-hydroxy-ethoxy)-amide (Compound 1) or a pharmaceutically acceptable salt thereof is administered to said mammal.

69. 69. The method of claim 68, wherein the RAS or RAF mutant cancer is, for example, a KRAS mutant cancer, an NRAS mutant cancer, an HRAS mutant cancer, or a BRAF mutant cancer.

70. 70. The method of claim 68 or 69, wherein the RAS mutant cancer is pancreatic cancer, colorectal cancer, lung cancer, melanoma, acute myeloid leukemia, bladder cancer, or head and neck cancer.

71. The method of claims 68 to 70, wherein the cancer is an NRAS mutant cancer.

72. 72. The method of claim 71, wherein the NRAS mutant cancer is NRAS mutant melanoma.

73. 73. The method of claim 72, wherein KRAS comprises a mutation at one or more positions selected from codons 12, 13, 59, and 61.

74. The method of claims 68 to 70, wherein the NRAS comprises a mutation at one or more positions selected from codons 12, 13, 59, 61 and 146.

75. 71. The method of claims 68 to 70, wherein the NRAS mutant form has a mutation at one or more amino acid positions selected from G12, G13, A59, Q61, K117 and A146.

76. 76. The method of claim 75, wherein the NRAS mutant form has one or more amino acid substitutions selected from G12C, G12R, G12S, G12A, G12D, G12V, G13C, G13R, G13S, G13A, G13D, G13V, A59D, A59T, Q61K, Q61L, Q61R, Q61H, K117N, K117R, K117E, A146P, A146T, and A146V.

77. The method of claims 68 to 70, wherein the cancer is an early, intermediate or advanced stage cancer, and the cancer may be locally advanced or metastatic.

78. The method of claims 68-70, wherein the mammal has previously undergone immunotherapy.

79. 79. The method of claim 78, wherein the mammal has previously undergone immunotherapy and is afflicted with NRAS mutant advanced melanoma.

80. 71. The method of claims 68 to 70, wherein the melanoma is selected from advanced melanoma, unresectable melanoma, metastatic melanoma, melanoma with a BRAF mutation, melanoma with an NRAS mutation, cutaneous melanoma, or intraocular melanoma.

81. 81. The method of claims 68 to 80, wherein compound 1 is in capsule form.

82. 82. The method of claim 81, wherein Compound 1 is administered at a dose of 5 to 50 mg once or twice daily.

83. 83. The method of claim 82, wherein Compound 1 is administered at a dose of 12 mg twice daily.

84. The method according to claims 68 to 83, wherein Compound 1 is any one of crystalline polymorphs of Form I to Form VI.

85. The method of claims 68 to 83, wherein compound 1 is a type I crystal.

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