Crystal form of pyrimido azacyclo-compound and preparation method and application of crystal form of pyrimido azacyclo-compound

Abstract

The invention relates to a crystal form of a pyrimido azacyclo compound as well as a preparation method and application of the crystal form. Specifically, the invention discloses crystal forms of a series of compounds as shown in the formula I. The crystal forms have excellent stability. The invention also discloses a preparation method of the crystal form and application of the crystal form in prevention and / or treatment of tumors.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a crystal form of a pyrimidine-azo-heterocyclic compound, its preparation method, and its application. Background Technology

[0002] The cell cycle refers to the entire process a cell undergoes from the completion of one division to the end of the next, divided into two phases: interphase and mitotic phase. Interphase is further divided into three phases: pre-DNA synthesis (G1 phase), DNA synthesis (S phase), and post-DNA synthesis (G2 phase); mitotic phase is the M phase. Cyclins and cyclin-dependent kinases (CDKs) are core molecules in the entire cell cycle regulation mechanism. At least 16 mammalian cyclins have been identified, among which cyclin B / CDK1 (Cyclin B / CDK1), cyclin A / CDK2 (Cyclin A / CDK2), cyclin E / CDK2 (Cyclin E / CDK2), cyclin D / CDK4 (Cyclin D / CDK4), cyclin D / CDK6 (Cyclin D / CDK6), and possibly other heterodynes are important regulators of cell cycle progression. Other functions of cyclin / CDK heterodynes include transcriptional regulation, DNA repair, differentiation, and apoptosis.

[0003] Cell cycle dysregulation has been confirmed as a common feature of human cancers, and cyclin-dependent kinase (CDK) inhibitors play a crucial role in cell cycle regulation, showing broad application prospects in cancer treatment. For example, CDK4 / 6 inhibitors palbociclib, ribociclib, and abemaciclib are used to treat breast cancer and other cancers; however, like other kinase inhibitors, their effectiveness may be limited over time by the development of primary or acquired resistance. Therefore, Pfizer's PF-06873600, as a novel multi-target CDK kinase inhibitor, holds promise for overcoming drug resistance under current conditions. PF-06873600 has also entered Phase I clinical trials for the treatment of metastatic breast cancer.

[0004] As one of the most promising areas of cancer treatment, cyclin-dependent kinase (CDK) inhibitors have made the development of new compounds with CDK kinase inhibitory activity and better pharmacodynamic and pharmacokinetic properties an important research project for the development of novel anti-tumor drugs, which will ultimately be used to treat human tumors and other diseases.

[0005] Patent publication number (WO 2022258023) discloses a series of compounds used as CDK kinase inhibitors, which exhibit very good CDK inhibitory activity. This patent application discloses a compound with the structural formula shown in Formula I.

[0006]

[0007] Currently, no reports have been found regarding the crystal form of this compound. Screening and developing crystal forms for the compound represented by Formula I to find stable and reliable crystal forms will ensure the quality stability of the compound, enabling better application in pharmaceuticals, drug formulations, and future clinical use. This will achieve stable and controllable drug quality, better dissolution, and higher bioavailability, thereby avoiding safety issues such as side effects caused by toxic impurities due to drug instability. This will be of great significance for future advancements. Summary of the Invention

[0008] The purpose of this invention is to provide a crystal form of a pyrimidine-azo-heterocyclic compound, its preparation method, and its application.

[0009] In a first aspect, the present invention provides a crystal form of a compound of formula I, said crystal form being a free base crystal form A.

[0010]

[0011] The X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.5±0.2°, and 24.3±0.2°.

[0012] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 9.6±0.2°, 14.4±0.2°, 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.2±0.2°, 20.5±0.2°, 24.3±0.2°, 25.1±0.2°, and 27.3±0.2°.

[0013] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 9.6±0.2°, 14.4±0.2°, 14.5±0.2°, 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.2±0.2°, 20.5±0.2°, 20.9±0.2°, 21.6±0.2°, 22.0±0.2°, 24.3±0.2°, 24.6±0.2°, 25.1±0.2°, and 27.3±0.2°.

[0014] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 8.2±0.2°, 9.6±0.2°, 10.0±0.2°, 11.0±0.2°, 11.4±0.2°, 12.5±0.2°, 12.9±0.2°, 13.5±0.2°, 14.4±0.2°, 14.5±0.2°, 15.2±0.2°, 16.1±0.2°, 17.0±0.2°, 17.5±0.2°, 18.4±0.2°, 19.0±0.2°, 20.2±0.2°, 20.5±0.2°, 20.9±0.2°, 21.6±0.2°, 2 2.0±0.2°, 23.0±0.2°, 24.3±0.2°, 24.6±0.2°, 25.1±0.2°, 25.9±0.2°, 26.9±0.2°, 27.3±0.2°, 27.7±0.2°, 28.2±0.2°, 28.6±0.2°, 29.0±0.2°, 29.5±0.2°, 30.5±0.2°, 31.3±0.2°, 32.3±0.2°, 32.8±0.2°, 33.5±0.2°, 34.0±0.2°, 34.6±0.2°, 35.6±0.2°, 36.1±0.2°, 36.7±0.2°, 38.6±0.2°.

[0015] In another preferred embodiment, the 2θ value of the X-ray powder diffraction pattern of the free alkali crystal form A has a deviation of ±0.3, more preferably ±0.2, and even more preferably ±0.1.

[0016] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form A is essentially as follows: Figure 2 As shown.

[0017] In another preferred embodiment, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the free alkali crystal form A are essentially as follows: Figure 3 As shown.

[0018] In another preferred embodiment, the free alkali crystal form A is a monoclinic crystal system, space group P21.

[0019] In another preferred embodiment, the unit cell parameters of the free alkali crystal form A are:

[0020] In another preferred embodiment, the free alkali crystal form A is an anhydrous compound.

[0021] In another preferred embodiment, the free alkali crystal form A has one or more features selected from the group consisting of:

[0022] 1) The free alkali crystal form A has a moisture absorption weight gain of ≤0.1% at 25℃ / 80%RH, preferably ≤0.09%, and more preferably ≤0.08%;

[0023] 2) At 37℃, the dynamic solubility of the free alkali crystal form A in H2O is 0.08-0.1 mg / mL;

[0024] 3) At 37℃, the dynamic solubility of the free alkali crystal form A in SGF is 0.28-0.30 mg / mL;

[0025] 4) At 37℃, the dynamic solubility of the free alkali crystal form A in FaSSIF is 0.09-0.1 mg / mL;

[0026] 5) At 37℃, the dynamic solubility of the free alkali crystal form A in FeSSIF is 0.13-0.14 mg / mL.

[0027] A second aspect of the present invention provides a method for preparing the crystal form described in the first aspect of the present invention, the method comprising the following steps:

[0028] The compound shown in Formula I is mixed with solvent m and stirred at room temperature for 1-10 days (preferably 3-7 days, more preferably 4-5 days), filtered, and dried to obtain the crystal form described in the first aspect of the present invention.

[0029] The solvent m is selected from the group consisting of chloroform, 2-methyltetrahydrofuran, petroleum ether, ethyl acetate, toluene, water, N,N-dimethylformamide, ethanol, or combinations thereof.

[0030] In another preferred embodiment, the compound represented by Formula I is amorphous.

[0031] In another preferred embodiment, the solvent m is a mixture of petroleum ether and ethyl acetate.

[0032] In another preferred embodiment, the solvent m is a mixture of petroleum ether and ethyl acetate, wherein the volume ratio of petroleum ether to ethyl acetate is 10-20, more preferably 14-16, and even more preferably 15.

[0033] In another preferred embodiment, the room temperature is 10-40°C, more preferably 15-35°C, and even more preferably 20-30°C.

[0034] In another preferred embodiment, the crystal form described in the first aspect of the present invention is prepared as follows:

[0035] The compound shown in Formula I is mixed with solvent n and stirred until completely dissolved. Solvent O is gradually added dropwise while stirring until a solid precipitates out, thus obtaining the crystal form described in the first aspect of the present invention.

[0036] The solvent n is selected from the group consisting of: acetone, tetrahydrofuran, acetonitrile, 1,4-dioxane, and ethyl acetate;

[0037] The solvent O is selected from the group consisting of: water, n-heptane, n-pentane, methyl tert-butyl ether, and petroleum ether.

[0038] In another preferred embodiment, the crystal form described in the first aspect of the present invention is prepared as follows:

[0039] Provide a first container and a second container;

[0040] The first container is open and contains the compound of Formula I and solvent n', wherein the compound of Formula I is dissolved in solvent n';

[0041] The second container contains solvent O';

[0042] The first container is placed inside the second container, and the second container is sealed. The solid in the first container precipitates out, thus obtaining the crystal form described in the first aspect of the present invention.

[0043] The solvent n' is selected from the group consisting of: acetone, tetrahydrofuran, acetonitrile, 1,4-dioxane, and ethyl acetate;

[0044] The solvent O' is selected from the group consisting of: water, n-heptane, n-pentane, methyl tert-butyl ether, and petroleum ether.

[0045] In another preferred embodiment, the compound of formula I in the first container is the crystal form described in the first aspect of the present invention.

[0046] A third aspect of the present invention provides a pharmaceutical composition comprising a safe and effective amount of the crystal form and a pharmaceutically acceptable carrier described in the first aspect of the present invention.

[0047] A fourth aspect of the present invention provides the use of the crystal form described in the first aspect of the present invention for the preparation of a pharmaceutical remedy, the pharmaceutical remedy being used for purposes selected from the group consisting of:

[0048] 1) Treat diseases or conditions related to CDK regulation;

[0049] 2) Treatment of tumors;

[0050] 3) Inhibits cell proliferation.

[0051] In another preferred embodiment, the tumor is a CDK-related tumor.

[0052] In another preferred embodiment, the tumor is a tumor that highly expresses CDK2, CDK4 and / or CDK6.

[0053] In another preferred embodiment, the tumor is selected from the group consisting of: breast cancer, ovarian cancer, bladder cancer, uterine cancer, lung cancer, colorectal cancer, prostate cancer, pancreatic cancer, gastric cancer, thyroid cancer, esophageal cancer, kidney cancer, liver cancer, head and neck glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia (CML), and acute myeloid leukemia (AML).

[0054] In another preferred embodiment, the lung cancer is non-small cell lung cancer.

[0055] In another preferred embodiment, the breast cancer is selected from the group consisting of: ductal carcinoma of the breast and squamous cell carcinoma of the breast.

[0056] In another preferred embodiment, the cells are cancer cells selected from the group consisting of: breast cancer, ovarian cancer, bladder cancer, uterine cancer, lung cancer, colorectal cancer, prostate cancer, pancreatic cancer, gastric cancer, thyroid cancer, esophageal cancer, kidney cancer, liver cancer, head and neck glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia (CML), and acute myeloid leukemia (AML).

[0057] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0058] Figure 1 This is the amorphous X-ray powder diffraction pattern of the present invention;

[0059] Figure 2 This is the X-ray powder diffraction pattern of the free alkali crystal form A of this invention;

[0060] Figure 3 This is the DSC-TGA image of the free alkali crystal form A of the present invention;

[0061] Figure 4 This is an X-ray powder diffraction overlay of the free alkali crystal form A of the present invention;

[0062] Figure 5 This is a single-crystal structure diagram of the free alkali crystal form A of the present invention;

[0063] Figure 6 This is a polarimetric micrograph (PLM) of the free alkali crystal form A of the present invention;

[0064] Figure 7 This is the X-ray powder diffraction pattern of the free alkali crystal form B of this invention;

[0065] Figure 8 This is the DSC-TGA image of the free alkali crystal form B of this invention;

[0066] Figure 9 This is a temperature-controlled X-ray powder diffraction overlay of the free alkali crystal form B of this invention;

[0067] Figure 10 This is the X-ray powder diffraction pattern of crystal form A of 1,5-naphthalene disulfonate of the present invention;

[0068] Figure 11 This is the DSC-TGA image of crystal form A of 1,5-naphthalene disulfonate of the present invention;

[0069] Figure 12 This is the X-ray powder diffraction pattern of methanesulfonate crystal form A of the present invention;

[0070] Figure 13 This is the DSC-TGA diagram of the methanesulfonate crystal form A of this invention;

[0071] Figure 14 This is the X-ray powder diffraction pattern of ethanesulfonate crystal form A of the present invention;

[0072] Figure 15 This is the DSC-TGA diagram of ethanesulfonate crystal form A of the present invention;

[0073] Figure 16 This is an X-ray powder diffraction pattern of the amorphous sample of the present invention after stirring in 2-methyltetrahydrofuran for 7 days;

[0074] Figure 17 This is an X-ray powder diffraction pattern of the amorphous sample of this invention after stirring in ethanol for 7 days;

[0075] Figure 18 These are X-ray powder diffraction patterns of the free alkali crystal form A of this invention after being placed under different conditions;

[0076] Figure 19 This is the DVS diagram of the free alkali crystal form A of the present invention;

[0077] Figure 20 These are X-ray powder diffraction patterns before and after DVS of the free alkali crystal form A of this invention;

[0078] Figure 21 These are X-ray powder diffraction patterns of the free alkali crystal type A before and after compression in this invention;

[0079] Figure 22 This is a solubility curve of the free alkali crystal form A of the present invention;

[0080] Figure 23 This is an X-ray powder diffraction overlay of a sample showing the dynamic solubility of free alkali crystal form A in water according to the present invention;

[0081] Figure 24 This is an X-ray powder diffraction overlay of a sample showing the dynamic solubility of free alkali crystal form A in SGF according to the present invention;

[0082] Figure 25 This is an X-ray powder diffraction overlay of a sample showing the dynamic solubility of free alkali crystal form A in FaSSIF according to the present invention;

[0083] Figure 26 This is an X-ray powder diffraction overlay of a sample showing the dynamic solubility of free alkali crystal form A in FeSSIF according to the present invention. Detailed Implementation

[0084] Through long-term and in-depth research, the inventors unexpectedly prepared a free base crystal form A with high purity, excellent solubility, excellent stability (crystal form stability, solid-state stability, pressure stability, etc.), excellent hygroscopicity, and excellent pharmacokinetic properties. This crystal form exhibits excellent drug-like properties, which is of great significance for the commercial application of propulsion I compounds and for their early accessibility to patients. Based on this, the inventors completed this invention.

[0085] the term

[0086] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0087] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0088] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0089] As used in this article, the term "n or more 2θ values ​​selected from the following group" refers to any positive integer including n and greater than n (e.g., n, n+1, ...), where the upper limit Nup is the number of all 2θ peaks in the group. For example, "3 or more" includes not only the positive integers of the upper limit Nup (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, ...), but also ranges such as "4 or more", "5 or more", and "6 or more".

[0090] Crystal form

[0091] Crystal form refers to the solid state of a drug. Drug crystal form research is essentially the study of the fundamental state of a drug. Only with a sufficient and comprehensive understanding of the crystal forms of chemical drugs can we find more suitable solid crystal forms for treating diseases. Drug crystal form can affect the physicochemical properties of drugs, directly influencing the basis for their clinical therapeutic effects. Different crystal forms of the same drug may exhibit significant differences in appearance, solubility, melting point, dissolution rate, and bioavailability, thus affecting the drug's stability, bioavailability, and efficacy. Therefore, studying the stable crystal form of a compound is of great significance.

[0092] The active ingredient of a drug generally exists in two or more crystalline forms, known as drug polymorphs. Different polymorphs have different solubilities and dissolution rates, affecting the drug's clinical therapeutic effect by causing changes in bioavailability in the body. Differences in drug polymorphs may affect its dissolution and absorption in the body, thus impacting bioavailability, clinical efficacy, and safety. Simultaneously, the stability of drug polymorphs is also crucial. To improve drug bioavailability, reduce toxicity, and enhance therapeutic efficacy, greater emphasis must be placed on drug polymorph stability. Stable polymorphs ensure the physicochemical stability of the drug dosage form during preparation and storage, maintaining good solubility and bioavailability, and ensuring equivalence between batches of the drug. The same drug often has multiple polymorphs; currently, the polymorph with better therapeutic effects and most suitable for clinical use is called the dominant drug polymorph.

[0093] This invention screened the crystal forms of the compound represented by Formula I, identifying as many different crystal forms of the active pharmaceutical ingredient as possible. The screened crystal forms were identified using powder X-ray diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), 1H NMR, and high-performance liquid chromatography (HPLC). Further thermodynamic stability and hygroscopicity studies determined the dominant free base crystal form A, providing a reference for crystal form selection in subsequent pharmacokinetic and animal experiments.

[0094] Further pharmacokinetic studies have shown that the free base crystal form A, represented by Formula I, provided by this invention is easily absorbed and highly stable in vivo, exhibiting excellent blood drug concentration distribution and high bioavailability. Furthermore, its good physical stability is highly beneficial for quality control in subsequent formulation development and allows for a longer shelf life, thereby enabling the drug to achieve better therapeutic effects.

[0095] In another preferred embodiment, the compound of formula I further has a crystal form selected from the group consisting of: free base crystal form B, 1,5-naphthalene disulfonate crystal form A, methanesulfonate crystal form A, and ethanesulfonate crystal form A;

[0096] The X-ray powder diffraction pattern of the free alkali crystal form B has characteristic peaks at the following 2θ values: 5.8±0.2°, 9.9±0.2°, 17.1±0.2°, 19.8±0.2°, and 20.4±0.2°.

[0097] The X-ray powder diffraction pattern of the 1,5-naphthalene disulfonate crystal form A has characteristic peaks at the following 2θ values: 6.0±0.2°, 17.3±0.2°, 17.9±0.2°, 20.8±0.2°, and 25.3±0.2°.

[0098] The X-ray powder diffraction pattern of the methanesulfonate crystal form A has characteristic peaks at the following 2θ values: 14.4±0.2°, 18.3±0.2°, 19.7±0.2°, 20.2±0.2°, and 20.7±0.2°.

[0099] The X-ray powder diffraction pattern of the ethanesulfonate crystal form A has characteristic peaks at the following 2θ values: 11.9±0.2°, 15.2±0.2°, 18.2±0.2°, 18.9±0.2°, and 21.4±0.2°.

[0100] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form B has characteristic peaks at the following 2θ values: 5.8±0.2°, 9.9±0.2°, 11.4±0.2°, 15.4±0.2°, 17.1±0.2°, 18.9±0.2°, 19.2±0.2°, 19.8±0.2°, 20.4±0.2°, and 25.7±0.2°.

[0102] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form B has characteristic peaks at the following 2θ values: 5.8±0.2°, 9.9±0.2°, 11.4±0.2°, 12.8±0.2°, 13.1±0.2°, 15.4±0.2°, 17.1±0.2°, 18.9±0.2°, 19.2±0.2°, 19.8±0.2°, 20.4±0.2°, 22.2±0.2°, 24.2±0.2°, 25.7±0.2°, 26.2±0.2°, 28.0±0.2°, and 30.0±0.2°.

[0103] In another preferred embodiment, the X-ray powder diffraction pattern of the free alkali crystal form B is substantially as follows: Figure 7 As shown.

[0104] In another preferred embodiment, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the free alkali crystal form B are essentially as follows: Figure 8 As shown.

[0105] In another preferred embodiment, the free alkali crystal form B is a hydrate.

[0106] In another preferred embodiment, the X-ray powder diffraction pattern of the 1,5-naphthalene disulfonate crystal form A has characteristic peaks at the following 2θ values: 6.0±0.2°, 17.0±0.2°, 17.3±0.2°, 17.9±0.2°, 18.1±0.2°, 20.8±0.2°, 21.0±0.2°, 22.5±0.2°, 25.3±0.2°, and 25.5±0.2°.

[0107] In another preferred embodiment, the X-ray powder diffraction pattern of the 1,5-naphthalene disulfonate crystal form A has characteristic peaks at the following 2θ values: 6.0±0.2°, 6.9±0.2°, 8.9±0.2°, 12.0±0.2°, 13.2±0.2°, 14.1±0.2°, 17.0±0.2°, 17.3±0.2°, 17.9±0.2°, 18.1±0.2°, 18.9±0.2°, 20.8±0.2°, 21.0±0.2°, 22.5±0.2°, 25.3±0.2°, 25.5±0.2°, 26.7±0.2°, and 27.5±0.2°.

[0108] In another preferred embodiment, the X-ray powder diffraction pattern of the 1,5-naphthalenedisulfonate crystal form A has characteristic peaks at the following 2θ values: 6.0±0.2°, 6.9±0.2°, 8.9±0.2°, 9.4±0.2°, 12.0±0.2°, 12.4±0.2°, 13.2±0.2°, 13.5±0.2°, 14.1±0.2°, 15.1±0.2°, 15.5±0.2°, 17.0±0.2°, 17.3±0.2°, 17.9±0.2°. ±0.2°, 18.1±0.2°, 18.9±0.2°, 20.8±0.2°, 21.0±0.2°, 22.1±0.2°, 22.5±0.2°, 24.4±0.2°, 25.3±0.2°, 25.5±0.2°, 26.2±0.2°, 26.7±0.2°, 27.5±0.2°, 28.6±0.2°, 29.2±0.2°, 29.9±0.2°, 34.4±0.2°, 38.8±0.2°.

[0109] In another preferred embodiment, the X-ray powder diffraction pattern of the 1,5-naphthalene disulfonate crystal form A is substantially as follows: Figure 10 As shown.

[0110] In another preferred embodiment, the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) spectra of the 1,5-naphthalene disulfonate crystal form A are substantially as follows: Figure 11 As shown.

[0111] In another preferred embodiment, in the 1,5-naphthalenedisulfonate crystal form A, the molar ratio of the compound of formula I to 1,5-naphthalenedisulfonic acid is 0.5-4, preferably 1-3.

[0112] In another preferred embodiment, the X-ray powder diffraction pattern of the methanesulfonate crystal form A has characteristic peaks at the following 2θ values: 14.4±0.2°, 14.7±0.2°, 17.1±0.2°, 17.9±0.2°, 18.3±0.2°, 19.7±0.2°, 20.2±0.2°, 20.7±0.2°, 21.2±0.2°, and 24.0±0.2°.

[0113] In another preferred embodiment, the X-ray powder diffraction pattern of the methanesulfonate crystal form A has characteristic peaks at the following 2θ values: 13.2±0.2°, 14.0±0.2°, 14.4±0.2°, 14.7±0.2°, 17.1±0.2°, 17.9±0.2°, 18.3±0.2°, 19.7±0.2°, 20.2±0.2°, 20.7±0.2°, 21.2±0.2°, 22.8±0.2°, 24.0±0.2°, 26.6±0.2°, 28.6±0.2°, and 29.3±0.2°.

[0114] In another preferred embodiment, the X-ray powder diffraction pattern of the methanesulfonate crystal form A has characteristic peaks at the following 2θ values: 7.2±0.2°, 9.6±0.2°, 13.2±0.2°, 13.5±0.2°, 14.0±0.2°, 14.4±0.2°, 14.7±0.2°, 17.1±0.2°, 17.9±0.2°, 18.3±0.2°, 19.0±0.2°, 19.7±0.2°, 20.2±0.2°, 20.7±0.2°, 21.2±0.2°, 21.8±0.2°. 0.2°, 22.8±0.2°, 23.2±0.2°, 23.8±0.2°, 24.0±0.2°, 24.7±0.2°, 25.9±0.2°, 26.6±0.2°, 27.6±0.2°, 28.6±0.2°, 29.3±0.2°, 30.3±0.2°, 31.0±0.2°, 31.6±0.2°, 32.1±0.2°, 32.8±0.2°, 33.4±0.2°, 35.0±0.2°, 35.8±0.2°, 37.3±0.2°.

[0115] In another preferred embodiment, the X-ray powder diffraction pattern of the methanesulfonate crystal form A is substantially as follows: Figure 12 As shown.

[0116] In another preferred embodiment, the thermogravimetric analysis (TGA) spectrum and differential scanning calorimetry (DSC) spectrum of the methanesulfonate crystal form A are substantially as follows: Figure 13 As shown.

[0117] In another preferred embodiment, in the methanesulfonate crystal form A, the molar ratio of the compound of formula I to methanesulfonic acid is 0.3-3, preferably 0.5-2.

[0118] In another preferred embodiment, the X-ray powder diffraction pattern of the ethanesulfonate crystal form A has characteristic peaks at the following 2θ values: 11.9±0.2°, 14.8±0.2°, 15.2±0.2°, 17.4±0.2°, 17.8±0.2°, 18.0±0.2°, 18.2±0.2°, 18.9±0.2°, 21.4±0.2°, and 23.8±0.2°.

[0119] In another preferred embodiment, the X-ray powder diffraction pattern of the ethanesulfonate crystal form A has characteristic peaks at the following 2θ values: 11.9±0.2°, 14.8±0.2°, 15.2±0.2°, 15.6±0.2°, 16.9±0.2°, 17.4±0.2°, 17.8±0.2°, 18.0±0.2°, 18.2±0.2°, 18.9±0.2°, 20.7±0.2°, 21.4±0.2°, 23.8±0.2°, 26.3±0.2°, and 27.8±0.2°.

[0120] In another preferred embodiment, the X-ray powder diffraction pattern of the ethanesulfonate crystal form A has characteristic peaks at the following 2θ values: 7.8±0.2°, 8.6±0.2°, 9.5±0.2°, 11.9±0.2°, 13.5±0.2°, 14.0±0.2°, 14.8±0.2°, 15.2±0.2°, 15.6±0.2°, 16.9±0.2°, 17.4±0.2°, 17.8±0.2°, 18.0±0.2°, 18.2±0.2°, 18.9±0.2°, 19.2±0.2°, 19.7±0.2°, 20.7±0.2°. 0.2°, 21.0±0.2°, 21.4±0.2°, 21.8±0.2°, 22.4±0.2°, 23.0±0.2°, 23.3±0.2°, 23.8±0.2°, 24.5±0.2°, 25.2±0.2°, 26.0±0.2°, 26.3±0.2°, 26.6±0.2°, 27.8±0.2°, 28.2±0.2°, 29.0±0.2°, 29.4±0.2°, 30.2±0.2°, 31.5±0.2°, 32.2±0.2°, 33.7±0.2°, 36.0±0.2°.

[0121] In another preferred embodiment, the X-ray powder diffraction pattern of the ethanesulfonate crystal form A is substantially as follows: Figure 14 As shown.

[0122] In another preferred embodiment, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the ethanesulfonate crystal form A are essentially as follows: Figure 15 As shown.

[0123] In another preferred embodiment, in the ethanesulfonate crystal form A, the molar ratio of the compound of formula I to ethanesulfonic acid is 0.3-3, preferably 0.5-2.

[0124] The embodiments of this invention describe in more detail the method for preparing the crystal form of the compound of formula I, but these specific methods do not constitute any limitation on this invention. The crystal form of the compound of this invention can also be conveniently obtained by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art.

[0125] It should be understood that, unless otherwise specified, the raw materials and reagents used in the preparation process of the crystal form of the compounds of the present invention can be purchased commercially.

[0126] Pharmaceutical composition

[0127] The present invention also provides a pharmaceutical composition comprising a safe and effective amount of the crystal form and a pharmaceutically acceptable carrier.

[0128] "Safe and effective dose" means that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects.

[0129] Typically, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, more preferably 10-200 mg of active ingredient per dose. Preferably, "one dose" refers to one tablet or one syringe.

[0130] "Pharmaceutical acceptable carriers" refer to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0131] "Compatibility" here refers to the ability of each component in the composition to interact with and incorporate with the active ingredient of the present invention without significantly reducing the efficacy of the active ingredient.

[0132] Pharmaceutically acceptable examples of carrier components include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0133] There are no particular limitations on the administration of the active ingredients or pharmaceutical compositions of the present invention. Representative administration methods include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous) administration.

[0134] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0135] In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or mixed with one or more of the following components:

[0136] (a) Fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol and silica;

[0137] (b) Adhesives, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0138] (c) Moisturizers, such as glycerin;

[0139] (d) Disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginate, certain complex silicates, and sodium carbonate;

[0140] (e) Slow solvents, such as paraffin;

[0141] (f) Absorption accelerators, such as quaternary ammonium compounds;

[0142] (g) Wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0143] (h) Adsorbent, for example, kaolin; and / or

[0144] (i) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof.

[0145] Buffers may also be included in the dosage forms of capsules, tablets, and pills.

[0146] The solid dosage form can also be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active ingredient from this composition can be delayed in a portion of the digestive tract. Examples of suitable encapsulating components are polymers and waxes.

[0147] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0148] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0149] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0150] When using the pharmaceutical composition, a safe and effective amount of the crystal form of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 20–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0151] The crystal form of the compound of the present invention can be administered alone or in combination with other therapeutic agents (such as antitumor drugs).

[0152] The crystalline form of compound (I) can also be used in combination with other known drugs for treating or improving similar symptoms. When used in combination, the original drug's administration method and dosage remain unchanged, while the crystalline form of compound (I) is taken simultaneously or subsequently. When the crystalline form of compound (I) is taken concurrently with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the crystalline form of compound (I) is preferred. Drug combination also includes taking the crystalline form of compound (I) with one or more other known drugs during overlapping time periods. When the crystalline form of compound (I) is used in combination with one or more other drugs, the dosage of the crystalline form of compound (I) or the known drug may be lower than the dosage when they are used alone.

[0153] Compared with the prior art, the present invention has the following main advantages:

[0154] (1) Compared with the amorphous form, the free alkali crystal form A has excellent chemical stability, solid-state stability and solution stability;

[0155] (2) The free alkali crystal form A of the present invention has excellent pharmacokinetic properties and good in vivo exposure in animals;

[0156] (3) The free alkali crystal form A of the present invention has the advantages of high purity, excellent solubility, excellent stability (crystal form stability, solid stability, pressure stability, etc.), and excellent hygroscopicity.

[0157] (4) The free alkali crystal form A of the present invention has better dissolution and higher bioavailability;

[0158] (5) The free alkali crystal form A of the present invention has the advantages of good safety and good effectiveness;

[0159] (6) The free alkali crystal form A of the present invention has excellent drug-forming properties;

[0160] (7) Compared with the acid salt crystal form, the free base crystal form A has stable chemical properties during the preparation process.

[0161] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0162] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0163] The technical solution of the present invention will be described in detail below.

[0164] The solvents and their corresponding Chinese names are shown in Table 1.

[0165] Table 1. Comparison of Chinese and English names of solvents used in the experiment

[0166]

[0167]

[0168] Crystal form analysis methods

[0169] 1.1 X-ray Powder Diffraction (XRPD)

[0170] XRPD images were acquired using an X-ray powder diffractometer manufactured by PANalytacal, and the scanning parameters are shown in Table 2.

[0171] Table 2 XRPD Test Parameters

[0172]

[0173] 1.2 Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

[0174] The TGA and DSC plots were acquired using a TA 5500 thermogravimetric analyzer and a TA 2500 differential scanning calorimeter, respectively. Table 3 lists the test parameters.

[0175] Table 3 DSC and TGA Test Parameters

[0176]

[0177]

[0178] 1.3 Tableting Machine

[0179] The tablet press used for the pressure stability assessment was a Xinno SYP-SBS manual tablet press.

[0180] 1.4 Dynamic Moisture Adsorption (DVS)

[0181] Dynamic moisture adsorption (DVS) curves were acquired using the DVSIntrinsic Plus of SMS (Surface Measurement Systems). Relative humidity at 25°C was corrected for the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS test parameters are listed in Table 4.

[0182] Table 4 DVS Test Parameters

[0183]

[0184] 1.5 High Performance Liquid Chromatography (HPLC)

[0185] Purity, dynamic solubility, and stability tests were performed using an Agilent 1260 high-performance liquid chromatograph, and the analytical conditions are shown in Table 5.

[0186] Table 5 HPLC test conditions

[0187]

[0188]

[0189] Regarding crystal form examples: Crystal form example 1: Preparation of amorphous form

[0190] The preparation method of compound I is the same as that in patent WO 2022258023, and the synthetic route is as follows:

[0191]

[0192] The specific steps are as follows:

[0193] Synthesis of Formula I

[0194] Compound int3 (80 mg) was dissolved in DMSO, and int7 (89 mg, 2.0 eq) and triethylamine (0.11 mL, 3.0 eq) were added. The mixture was reacted overnight at 60 °C. After the reaction was monitored to complete, water was added to the system, followed by extraction with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by preparative blotting to obtain the product (50 mg). 1H NMR (400MHz, CDCl3) δ8.45(s,1H),7.36(s,1H),6.77(t,J=75.2Hz,1H),5.82(s,1H),5.57(d,J=55.6Hz,1H),3.89–3.73(m,2H),2.91(m,J=12.0, 2.8Hz,2H),2.83(s,3H),2.74(s,1H),2.34–2.11(m,4H),2.10–1.98(m, 2H),1.99–1.78(m,2H),1.78–1.58(m,2H),1.16(s,3H).MS[M+1]:488.5.

[0195] The solid of formula I prepared by the above method was characterized by XRPD, and the results are as follows: Figure 1 As shown, the obtained solid is amorphous.

[0196] Crystal Form Example 2: Preparation of Free Base Crystal Form A

[0197] The above amorphous solid was pulped with a petroleum ether / ethyl acetate (15:1) mixture, filtered, and dried. The resulting solid was characterized by XRPD, and the results are as follows: Figure 2 As shown, the obtained solid is free alkali crystal type A, and its X-ray powder diffraction peak data are shown in Table 6.

[0198] Table 6

[0199]

[0200]

[0201] Thermogravimetric analysis and differential scanning calorimetry (TGA-DSC) of free alkali crystal form A are as follows: Figure 3 As shown.

[0202] The obtained free alkali crystal form A was used as a raw material in the crystal form preparation and salt form preparation experiments described below. The solubility of free alkali crystal form A in common solvents at room temperature was evaluated by visual inspection, and the results are shown in Table 7.

[0203] Table 7. Preliminary solubility results of free alkali crystal form A

[0204] Serial Number solvent Solubility (mg / mL) 1 MeOH 19.0<S<38.0 2 EtOH 1.9<S<4.75 3 IPA S<2.0 4 Acetone S>40.0 5 MIBK 5.75<S<11.5 6 EtOAc 5.0<S<10.0 7 IPAC 2.2<S<5.5 8 MTBE S<2.1 9 THF S>44.0 10 2-MeTHF 10.5<S<21.0 11 1,4-dioxane 22.0<S<44.0 12 ACN 23.0<S<46.0 13 <![CDATA[CHCl3]]> 10.0<S<20.0 14 DCM 20.0<S<40.0 15 n-heptane S<2.0 16 Toluene S<2.0 17 DMAc S>38.0 18 DMSO S>42.0 19 NMP S>40.0 20 <![CDATA[H2O]]> S<1.9

[0205] S: Solubility.

[0206] 20.4 mg of free alkali crystal form A was weighed and placed in a 3 mL glass bottle. 2.0 mL of ethyl acetate was added, and the sample was sonicated appropriately to promote dissolution. The solution was then filtered, and the filtrate was transferred to a clean 3 mL glass bottle. This bottle was then placed open into a 20 mL glass bottle pre-filled with 4 mL of n-pentane, and the 20 mL bottle was sealed. Finally, the system was placed in a fume hood for gas-liquid diffusion experiments to obtain rod-shaped crystals. The XRPD characterization results for this sample are shown in the overlay image below. Figure 4 As shown.

[0207] from Figure 4 It can be seen that the solid is a free alkali crystal type A.

[0208] The single crystal structure of the above samples is as follows: Figure 5 As shown, characterization reveals that this single crystal belongs to the monoclinic crystal system, space group P21, and its unit cell parameters are: The crystal lattice contains no water of crystallization molecules or organic solvent molecules, indicating that the free alkali crystal form A is an anhydrous hydrate. The polarimetric micrograph (PLM) of the obtained crystal is shown below. Figure 6 As shown.

[0209] Crystal Form Example 3: Preparation of Free Base Crystal Form B

[0210] Free alkali crystal form B was obtained by volatilization of free alkali crystal form A in EtOH / H2O (v:v, 4:1) at room temperature, as shown in the XRPD results. Figure 7 As shown, its X-ray powder diffraction peak data are shown in Table 8.

[0211] Table 8

[0212] 2θ(°)±0.2° Relative strength (%) 2θ(°)±0.2° Relative strength (%) 5.8 63.85 19.8 79.82 9.9 58.87 20.4 100.00 11.4 50.72 22.2 25.26 12.8 29.99 24.2 11.08 13.1 20.62 25.7 30.17 15.4 46.88 26.2 19.34 17.1 75.64 28.0 9.07 18.9 45.01 30.0 7.73 19.2 49.73

[0213] The TGA and DSC results of free alkali crystal form B are as follows: Figure 8 As shown, the TGA results indicate that the sample lost 6.78% of its weight when heated to 120℃; the DSC curves showed endothermic peaks (peak temperatures) at 73.7, 98.9, 196.7, and 283.7℃, and exothermic peaks (peak temperatures) at 166.4 and 183.9℃.

[0214] Free alkali crystal form B 1 No residual solvent EtOH was observed by 1H NMR.

[0215] Free alkali crystalline form B becomes amorphous after heating to 120℃ under nitrogen purging. Upon further heating to 180℃, it transforms into free alkali crystalline form A. No crystalline form change is observed after cooling to 30℃ or exposure to room humidity (the difference in XRPD diffraction peak positions at high and low temperatures is due to lattice thermal expansion and contraction). The temperature-dependent XRPD test results are as follows... Figure 9 As shown. Based on the temperature-dependent experimental data and NMR results of free alkali crystal form B, it is inferred that free alkali crystal form B is a hydrate.

[0216] "Room humidity" refers to the humidity of the laboratory environment, such as 10-50%.

[0217] Crystal Form Example 4: Preparation of Crystal Form A of 1,5-Naphthalenedisulfonate

[0218] 1,5-Naphthalenedisulfonate crystal form A is obtained by suspending and stirring free alkali crystal form A and 1,5-naphthalenedisulfonic acid in ethanol at a molar ratio of 1:1 at room temperature.

[0219] The XRPD results of 1,5-naphthalenedisulfonate crystal form A are as follows: Figure 10 As shown, its X-ray powder diffraction peak data are shown in Table 9.

[0220] Table 9

[0221]

[0222]

[0223] The TGA and DSC results of 1,5-naphthalenedisulfonate crystal form A are as follows: Figure 11 As shown, the TGA results indicate that the sample weight loss was 1.32% when the temperature was raised to 120℃; endothermic peaks were observed in the DSC curve at 69.8℃ and 213.8℃ (peak temperature).

[0224] 1 ¹H NMR showed a signal of 1,5-naphthalenedisulfonic acid in DMSO-d6 at a molar ratio of 0.5 to the compound, and no signal of residual solvent EtOH was observed.

[0225] Crystal Form Example 5: Preparation of Methanesulfonate Crystal Form A

[0226] Methanesulfonate crystal form A is obtained by suspending and stirring free base crystal form A and methanesulfonic acid in IPAc at a molar ratio of 1:1 overnight at room temperature to form a gel. The gel is then subjected to temperature cycling (50℃~5℃) for about 3 days.

[0227] The XRPD results of methanesulfonate crystal form A are as follows: Figure 12As shown, its X-ray powder diffraction peak data are shown in Table 10.

[0228] Table 10

[0229]

[0230]

[0231] The TGA and DSC results of methanesulfonate crystal form A are as follows: Figure 13 As shown, the TGA results indicate that the sample weight loss was 1.03% when heated to 100℃; the DSC curve showed an endothermic peak at 158.1℃ (peak temperature).

[0232] 1 The methanesulfonic acid signal was observed in DMSO-d6 by 1.0 molar ratio of the compound to 1.0 by 1H NMR, and no residual solvent IPAC signal was observed.

[0233] Crystal Form Example 6: Preparation of Ethylene Sulfonate Crystal Form A

[0234] Ethylene sulfonate crystal form A is obtained by suspending and stirring free alkali crystal form A and ethanesulfonic acid in IPAC at a molar ratio of 1:1 overnight at room temperature to form a gel. The gel is then subjected to temperature cycling (50℃~5℃) for about 3 days.

[0235] The XRPD results of ethanesulfonate crystal form A are as follows: Figure 14 As shown, its X-ray powder diffraction peak data are shown in Table 11.

[0236] Table 11

[0237] 2θ(°)±0.2° Relative strength (%) 2θ(°)±0.2° Relative strength (%) 7.8 3.53 21.4 81.37 8.6 2.69 21.8 8.55 9.5 4.81 22.4 9.13 11.9 70.18 23.0 6.85 13.5 4.10 23.3 6.87 14.0 5.32 23.8 17.33 14.8 17.22 24.5 8.97 15.2 43.39 25.2 3.86 15.6 10.36 26.0 6.51 16.9 12.51 26.3 11.72 17.4 19.33 26.6 2.97 17.8 15.46 27.8 11.01 18.0 17.96 28.2 3.39 18.2 22.74 29.0 2.15 18.9 100.00 29.4 3.12 19.2 9.85 30.2 3.90 19.7 6.96 31.5 2.62 20.7 11.97 32.2 3.27 21.0 7.17 33.7 2.49 36.0 1.71

[0238] The TGA and DSC results of ethanesulfonate crystal form A are as follows: Figure 15 As shown, the TGA results indicate that the sample weight loss was 1.61% when the temperature was raised to 120℃; the DSC curve showed an endothermic peak at 177.8℃ (peak temperature).

[0239] The 1H NMR of ethanesulfonate crystal form A showed a signal of ethanesulfonic acid in DMSO-d6 at a molar ratio of 1.0 to the compound, and no signal of residual solvent IPAC was observed.

[0240] Crystal form test example 1: Transformation relationship of free base crystal forms A / B

[0241] To investigate the interconversion relationship between amorphous form A and hydrated form B, a suspension competition experiment was conducted using the following steps:

[0242] 1. Prepare a suspension in the corresponding solvent for the sample of compound crystal form A, and equilibrate at room temperature for 4 hours;

[0243] 2. Weigh compound samples of crystal form A and crystal form B in a mass ratio of approximately 1:1 into an HPLC vial;

[0244] 3. Filter the suspension from step 1 through a PTFE membrane and add the filtrate to the corresponding HPLC vial from step 2;

[0245] 4. After suspending and stirring at room temperature for 4 days, take a wet sample, cover it with a membrane, and test XRPD.

[0246] The results are shown in Table 12. Under room temperature conditions and at all water activities, crystal form B will transform into crystal form A. Therefore, it can be seen that free base crystal form A is the stable crystal form at room temperature.

[0247] Table 12 Results of Hybrid Suspension Competition Test

[0248]

[0249] Crystal form test example 2: Transformation relationship between free base crystal form A and amorphous form

[0250] Weigh 20 mg of the amorphous sample and add it to a glass vial. Add 0.5 mL of 2-methyltetrahydrofuran and ethanol respectively. Stir the resulting suspension at room temperature for 7 days. Filter to collect the solid and perform XRPD testing. See [link to XRPD test]. Figure 16 and Figure 17 The test results are shown in the table below.

[0251] Table 13 Results of Hybrid Suspension Competition Test

[0252]

[0253] The results above show that amorphous samples are unstable and easily transform into free alkali crystal form A.

[0254] Crystal form test example 3: Solid-state stability assessment of free alkali crystal form A

[0255] As can be seen from Test Example 1 and Test Example 2, the free alkali crystal form A is a stable crystal form, and it will be further studied now.

[0256] The free alkali crystal form A sample was placed under the following conditions: 60℃ / closed / 1 day, 25℃ / 60%RH / open / 1 week, and 40℃ / 75%RH / open / 1 week. The physical and chemical stability of the sample was determined by XRPD and HPLC. The data are shown in Table 14, and the XRPD overlay is shown in [Table 14]. Figure 18 .

[0257] The results showed that the free alkali crystal form A sample did not experience a significant decrease in HPLC purity under any of the test conditions, and no crystal form change was observed.

[0258] Table 14 Summary of Solid State Stability Assessment

[0259]

[0260] Example 4: Hygroscopicity assessment of free alkali crystal form A

[0261] The hygroscopicity of the free alkali crystal form A sample was evaluated using DVS. The DVS test results and the XRPD results of the sample after DVS testing are shown in [Figure number missing]. Figure 19 and Figure 20 .

[0262] The hygroscopicity assessment results showed that the moisture absorption weight gain at 25℃ / 80%RH was 0.0799%, the sample had almost no hygroscopicity, and no obvious crystal form change was observed after the test.

[0263] Crystal form test example 5: Pressure stability of free alkali crystal form A

[0264] The free alkali crystal form A sample did not change its crystal form after being pressed at 350 MPa. XRPD results are shown below. Figure 21 .

[0265] Crystal form test example 6: Dynamic solubility of free base crystal form A

[0266] The dynamic solubility of free alkali crystalline form A in H2O, SGF, FaSSIF, and FeSSIF was tested at 37℃. Solid feed concentration of 10 mg / mL was used for rotary mixing at 37℃, and the solubility of each sample was measured at different time points (1, 2, 4, and 24 hours). After each time point, samples were centrifuged and filtered; the HPLC concentration and pH of the filtrate were measured, and the XRPD of the centrifuged solid samples was tested. The results of the dynamic solubility test are summarized in Table 15, and the solubility curves are shown in [Figure number missing]. Figure 22 .

[0267] The results showed that the solubility of free alkali crystal form A in SGF was 0.3 mg / mL, which was higher than that of other solvents. Its solubility in FaSSIF and FeSSIF was close to that in H2O, ranging from 0.1 to 0.2 mg / mL. The XRPD results for water, SGF, FaSSIF, and FeSSIF are shown in the table below. Figures 23 to 26 The solid did not undergo any crystal form change in any of the system tests.

[0268] Table 15 Summary of Dynamic Solubility Test Results for Free Alkali Crystal Form A

[0269]

[0270] S: Free state concentration (mg / mL); FC: Crystal form transformation; No: No crystal form transformation occurred.

[0271] Crystal form test example 7: Determination of the stability of acid salts

[0272] In the salt form screening test, potential salt form samples were obtained in the 1,5-naphthalenedisulfonic acid, methanesulfonic acid, and ethanesulfonic acid systems, respectively. HPLC tests were performed on these samples, and the results are shown in Table 16. Compared to the free alkali crystal form A, the acidic salt samples all showed significant impurities at RRT = 0.81 in HPLC.

[0273] To investigate the possible source of this impurity, LC-MS analysis was performed on the above samples. The results showed that the m / z value of the impurity was 471.1, suggesting that the impurity was generated by the dehydration of the free base sample under acidic conditions. The above experiments indicate that there is a risk of chemical instability during the preparation of salt-type samples.

[0274] Table 16. Percentage of Impurities in Acid Salt Degradation

[0275] Crystal form RRT 0.81 (%) Free alkali crystal form A - 1,5-Naphthalenedisulfonate crystal form A 2.65 Methanesulfonate crystal form A 0.5 Ethyl sulfonate crystal form A 0.09

[0276] "-" indicates that there is no peak area here.

[0277] Example 8: Pharmacokinetic determination of free base crystal form A

[0278] The free basal crystal form A of compound I was subjected to in vivo drug metabolism experiments in rats.

[0279] SD rats were administered free alkali crystal form A via single gavage. Blood samples were collected at different time points to determine the drug concentration in rat plasma after administration and to calculate relevant pharmacokinetic parameters.

[0280] Preparation of test sample solution

[0281] Accurately weigh an appropriate amount of the test sample, transfer it to a suitable container, add 5% DMSO and 10% Kolliphor HS15 (melted by heating with a magnetic stirrer at 50℃±5℃ or in a water bath at 50℃±5℃), and mix thoroughly. Then add an appropriate amount of sodium chloride injection, mix thoroughly, and prepare the solution to the required volume with sodium chloride injection. Mix thoroughly to obtain test sample solutions with concentrations of 0.1, 0.2, and 0.4 mg / mL, respectively.

[0282] Analysis of test solution

[0283] The prepared test solutions were analyzed by the analytical department using LC-MS / MS.

[0284] Animal reception and adaptation

[0285] Healthy male SD rats were used in this study and were fasted overnight.

[0286] Sample collection and processing

[0287] The blood collection time points are:

[0288] Oral administration: 5 min, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h after administration.

[0289] Pharmacokinetic Analysis

[0290] Plasma concentration data were analyzed using the pharmacokinetic data analysis software WinNonlin. The t-values ​​were calculated using the non-compartmental model (NCA). 1 / 2 C max 、T max And AUC, etc.

[0291] Experimental results

[0292] SD rats were given free alkali crystal form A via a single gavage at a dose of 2 mg / kg. The main pharmacokinetic parameters are shown in Table 17.

[0293] As can be seen from Table 17, the free alkali crystal form A has good exposure levels in animals.

[0294] Table 17

[0295] dose <![CDATA[C max ]]> <![CDATA[T max ]]> <![CDATA[t 1 / 2 ]]> <![CDATA[AUC last ]]> <![CDATA[AUC inf ]]> Vd / F Cl / F <![CDATA[MRT last ]]> (mg / kg) (ng / mL) (h) (h) (h·ng / mL) (h·ng / mL) (L / kg) (L / h / kg) (h) 2 155.88 0.25 4.70 624.20 680.22 19.79 2.95 4.92

[0296] In summary, the inventors conducted relevant crystal form evaluations on free alkali crystal form A, free alkali crystal form B, amorphous and acidic salt samples. The evaluation items included dynamic solubility, solid-state stability, hygroscopicity, grinding stability, and pressure stability. Based on the combined results of salt form screening, polymorphic form screening tests, sample characterization, and crystal form evaluation, free alkali crystal form A is a stable crystal form at room temperature and possesses good physicochemical properties, making it suitable for further development.

[0297] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A crystal form of a compound of formula I, characterized in that, The crystal form is free alkali crystal form A. The X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.5±0.2°, and 24.3±0.2°.

2. The crystal form as described in claim 1, characterized in that, The X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 9.6±0.2°, 14.4±0.2°, 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.2±0.2°, 20.5±0.2°, 24.3±0.2°, 25.1±0.2°, and 27.3±0.2°.

3. The crystal form as described in claim 1, characterized in that, The X-ray powder diffraction pattern of the free alkali crystal form A has characteristic peaks at the following 2θ values: 9.6±0.2°, 14.4±0.2°, 14.5±0.2°, 15.2±0.2°, 18.4±0.2°, 19.0±0.2°, 20.2±0.2°, 20.5±0.2°, 20.9±0.2°, 21.6±0.2°, 22.0±0.2°, 24.3±0.2°, 24.6±0.2°, 25.1±0.2°, and 27.3±0.2°.

4. The crystal form as described in claim 1, characterized in that, The X-ray powder diffraction pattern of the free alkali crystal form A is basically shown in Figure 2.

5. The crystal form as described in claim 1, characterized in that, The thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of the free alkali crystal form A are basically shown in Figure 3.

6. The crystal form as described in claim 1, characterized in that, The free alkali crystal form A is an anhydrous compound.

7. A method for preparing the crystal form according to claim 1, characterized in that, The method includes the following steps: The compound shown in Formula I is mixed with solvent m and stirred at room temperature for 1-10 days (preferably 3-7 days, more preferably 4-5 days), filtered, and dried to obtain the crystal form described in claim 1. The solvent m is selected from the group consisting of chloroform, 2-methyltetrahydrofuran, petroleum ether, ethyl acetate, toluene, water, N,N-dimethylformamide, ethanol, or combinations thereof.

8. A pharmaceutical composition, characterized in that, The crystal form and pharmaceutically acceptable carrier of claim 1 are contained in a safe and effective amount.

9. Use of the crystal form according to claim 1, characterized in that, Used to prepare a medicine, said medicine being used for purposes selected from the group consisting of: 1) Treat diseases or conditions related to CDK regulation; 2) Treatment of tumors; 3) Inhibits cell proliferation.

10. The use as described in claim 9, characterized in that, The tumors are selected from the following groups: breast cancer, ovarian cancer, bladder cancer, uterine cancer, lung cancer, colorectal cancer, prostate cancer, pancreatic cancer, gastric cancer, thyroid cancer, esophageal cancer, kidney cancer, liver cancer, head and neck cancer, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia (CML), and acute myeloid leukemia (AML).

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