A crystal form of a thienopyrimidine derivative methanesulfonate and a preparation method and application thereof

Abstract

The application discloses a crystal form of a thienopyrimidine derivative methanesulfonic acid salt and a preparation method and application thereof. The crystal form of the thienopyrimidine derivative methanesulfonic acid salt comprises a crystal form I, a crystal form II and a crystal form III, wherein the crystal form I is an anhydrous crystal form with a characteristic XRPD diffraction peak and a single melting point; the initial melting point is about 226 DEG C; and the physical and chemical stability is good within 2 weeks under high temperature (60 DEG C) and accelerated conditions (40 DEG C / 75%RH). The application further discloses a medicine / drug composition. The application further discloses application of the crystal form of the thienopyrimidine derivative methanesulfonic acid salt and the preparation method thereof in preparation of a medicine for treating diseases caused by abnormal cell proliferation caused by overexpression of EGFR. The application has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of chemical synthesis technology, specifically relating to the crystal form of a thiophene pyrimidine derivative methanesulfonate, its preparation method, and its application. Background Technology

[0002] Epidermal growth factor receptor (EGFR, Erbb1) is involved in the proliferation of both normal and malignant tumor cells. EGFR overexpression has been found to be associated with cancers such as lung cancer, breast cancer, head and neck cancer, and bladder cancer.

[0003] The EGFR family comprises four receptor tyrosine kinases: EGFR (ErbB1), human epidermal growth factor receptor 2 (HER2, ErbB2), HER3 (ErbB3), and HER4 (ErbB4). Tyrosine kinases play a role in signal transduction. They bind ligands to the receptor's ligand-binding domain extracellularly, forming active homodimers or heterodimers, leading to autophosphorylation of EGFR and activation of stopping proteins (Greulich H. Chen, PLoS Med, 2005, 2, e313; Olayioye MA, EMBOJ, 2000, 19, 3159-3167). These signals promote cellular processes such as proliferation, protein synthesis, angiogenesis, cell growth, and survival.

[0004] EGFR overexpression is present in approximately 70% of cancer patients (Seymour, LK, Curr Drug Targets, 2001, 2, 117-133). EGFR tyrosine kinase has become a therapeutic target, and several drugs have been developed to inhibit kinase activity and block its signal transduction pathway as ATP competitors, such as the FDA-approved Tarceva, Irressa, and Gilotrif (all three are 4-amino-quinazoline inhibitors). These drugs have been widely used in patients with EGFR-overexpressing non-small cell lung cancer (NSCLC), including wild-type and active mutation patients (W. Pao, Nat. Rev. Cancer, 2010, 10, 760-774; R. Rosell, Lancet Oncol, 2012, 13, 239-246; NULin, Breast Cancer Res, 2004, 6, 204-210).

[0005] Two common active mutations were found in patients: L858R and E746-A750 deletion. Mechanistic studies suggest that the clinical activity of Tarceva and Iressa in patients with active mutations may be a result of the combined effects of the enhanced binding affinity of the inhibitor to the mutant kinase and the addiction of mutant cells to proto-oncogenes (JA. Engelman, Science, 2007, 316, 1039-1043).

[0006] However, these first- and second-generation inhibitors with a 4-amino-quinazoline core structure are not effective in approximately 50% of patients with relapsed and acquired resistance diseases such as NSCLC. Acquired resistance is caused by mutations in the T790M gate residue (LV. Sequist, SciTransl Med, 2011, 3, 75ra26; S. Kobayashi, NEngl J Med, 2005, 352, 786-792; W. Pao, PLoS M Med, 2005, 2, 373; JA. Engelman, Semin Respir Crit Care Med, 2005, 26, 314-322). This mutation (the second mutation) increases the binding affinity of ATP for EGFR tyrosine kinase and affects the thermodynamic and kinetic binding properties of these formulations (CH. Yun, ProcNatlAcadSciUSA, 2008, 105, 2070-2075; CancerCell, 2007, 11, 217-227; M. Azam, NatStructMolBiol, 2008, 15, 1109-1118; TA. Carter, ProcNatlAcadSciUSA, 2005, 102, 11011-11016). The larger methionine side chain in the choke region prevents those drug molecules from interacting with the ATP binding pocket at clinically effective concentrations.

[0007] Second-generation covalent EGFR inhibitors, such as the FDA-approved afatinib and the clinical-tested compound HKI-272, are effective in patients with the T790M mutation. However, the use of afatinib in patients with active mutations is limited due to dose-limiting toxicity that causes inhibition of wild-type EGFR.

[0008] Thiophene pyrimidine derivative mesylate is a potent and selective inhibitor of mutant epidermal growth factor receptor (EGFR) tyrosine kinase without affecting wild-type EGFR, thus reducing side effects.

[0009] The thiophene pyrimidine derivative described in this invention, as a third-generation EGFR-TKI, also exhibits strong inhibitory activity against EGFR-19del and L858R, two EGFR-sensitive gene mutations. It also shows strong inhibitory activity against EGFR19del / T790M and EGFRL858R / T790M, two drug-resistant dual-gene mutations resulting from resistance to first- and second-generation EGFR inhibitors. Furthermore, this thiophene pyrimidine derivative demonstrates good selectivity for the aforementioned tumor mutation genes and EGFR wild-type. Therefore, this thiophene pyrimidine derivative exhibits good inhibitory activity against tumors with the aforementioned sensitive / resistant gene mutations, demonstrating a good tumor-suppressive effect; secondly, its low activity against EGFR wild-type reduces the side effects of the compound, alleviating patient suffering and improving patient compliance. Animal pharmacokinetic studies have shown that this thiophene pyrimidine derivative is orally absorbed, exhibiting varying bioavailability across different animal species and sexes, ranging from 40% to 60%. The thiophene pyrimidine derivative is distributed in tissues greater than in plasma, with the highest distribution observed in lung tissue.

[0010] Therefore, this thiophene pyrimidine derivative exhibits good tumor-suppressive activity, low side effects, and ideal pharmacokinetic performance. Based on these factors, there is an urgent need to develop stable, orally effective formulations of thiophene pyrimidine derivatives suitable for preclinical animal studies and human administration during clinical trials. Summary of the Invention

[0011] To address the shortcomings of existing technologies, the present invention aims to provide a crystal form of a thiophene pyrimidine derivative mesylate, its preparation method, and its applications. In drug development, there is a need to develop stable, orally effective formulations suitable for preclinical animal studies and human administration during clinical trials for oral medications. Through research and development of thiophene pyrimidine derivatives protected by patents (US10829495B2 / CN108289895B), the present invention has for the first time successfully synthesized the crystal form of the thiophene pyrimidine derivative mesylate, which is a product with a unique and stable crystal form and has been successfully used in preclinical and clinical studies.

[0012] This invention provides a crystal form of a thiophene pyrimidine derivative methanesulfonate, the structural formula of which is shown in formula (I):

[0013]

[0014] The crystal form of the thiophene pyrimidine derivative methanesulfonate is selected from the following group: crystal form I, crystal form II and crystal form III.

[0015] In some embodiments, the crystal form is crystal form I.

[0016] In some embodiments, the X-ray powder diffraction pattern of crystal form I of the thiophene pyrimidine derivative methanesulfonate has characteristic peaks at 2θ angles of 7.379, 9.535, 10.822, 11.859, 13.377, 15.575, 17.249, 18.664, 19.353, 20.072, 21.592, 21.906, 22.402, 23.644, 25.683, and 26.240.

[0017] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 17 The X-ray powder diffraction pattern shown.

[0018] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 18 The XRPD peak finding report shown.

[0019] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 19 The DSC and TGA spectra shown are shown.

[0020] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form I of the thiophene pyrimidine derivative methanesulfonate includes an endothermic peak at 228.66 °C.

[0021] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 20 shown 1 HNMR image.

[0022] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 21 The DSC graph shown is obtained after heating to 150°C.

[0023] In some embodiments, the crystal form I of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 22 The XRPD overlay images shown are before and after heating.

[0024] Specifically, the thiophene pyrimidine derivative methanesulfonate crystal form I is an anhydrous crystal form with a single melting point and characteristic XRPD diffraction peaks; the initial melting point is around 226°C; and it exhibits good physical and chemical stability within 2 weeks under high temperature (60°C) and accelerated conditions (40°C / 75% RH).

[0025] In some embodiments, the crystal form is crystal form II.

[0026] In some embodiments, the X-ray powder diffraction pattern of crystal form II of the thiophene pyrimidine derivative methanesulfonate has characteristic peaks at 2θ angles of 14.001, 18.076, 18.743, 20.312, 22.602, 24.038, and 25.686.

[0027] In some embodiments, the crystal form II of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 24 The X-ray powder diffraction pattern shown.

[0028] In some embodiments, the crystal form II of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 25 The XRPD peak finding report shown.

[0029] In some embodiments, the crystal form II of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 26 The DSC and TGA spectra shown are shown.

[0030] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form II of the thiophene pyrimidine derivative methanesulfonate includes an endothermic peak at 227.05 °C.

[0031] In some embodiments, the crystal form II of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 27 shown 1 HNMR image.

[0032] In some embodiments, the crystal form II of the thiophene pyrimidine derivative methanesulfonate has essentially the following characteristics: Figure 28 The image shown is an XRPD plot after heating to 150°C.

[0033] In some embodiments, the crystal form is crystal form III.

[0034] In some embodiments, the X-ray powder diffraction pattern of crystal form III of the thiophene pyrimidine derivative methanesulfonate has characteristic peaks at 2θ angles of 6.005, 12.020, 13.977, 18.305, 18.722, 20.602, 22.516, 24.096, and 25.807.

[0035] In some embodiments, the thiophene pyrimidine derivative methanesulfonate crystal form III has essentially the following properties: Figure 30 The X-ray powder diffraction pattern shown.

[0036] In some embodiments, the thiophene pyrimidine derivative methanesulfonate crystal form III has essentially the following properties: Figure 31 The XRPD peak finding report shown.

[0037] In some embodiments, the thiophene pyrimidine derivative methanesulfonate crystal form III has essentially the following properties: Figure 32 The DSC and TGA spectra shown are shown.

[0038] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form III of the thiophene pyrimidine derivative methanesulfonate includes an endothermic peak at 226.91 °C.

[0039] In some embodiments, the thiophene pyrimidine derivative methanesulfonate crystal form III has essentially the following properties: Figure 33 shown 1 HNMR image.

[0040] In some embodiments, the thiophene pyrimidine derivative methanesulfonate crystal form III has essentially the following properties: Figure 34 The image shown is an XRPD plot after heating to 150°C.

[0041] The present invention also provides a method for preparing crystal form I of the thiophene pyrimidine derivative methanesulfonate as described above, the preparation method comprising the following:

[0042] Method 1: Solution evaporation crystallization (EVA): Place the thiophene pyrimidine derivative methanesulfonate sample in a glass bottle, add solvent, sonicate to promote dissolution, filter through a 0.45-0.5 μm filter membrane into a new sample bottle, place the sample bottle open in a fume hood, and allow the solvent to evaporate naturally at room temperature. After a large amount of solid has precipitated, the resulting sample is crystal form I of the thiophene pyrimidine derivative methanesulfonate; preferably, a 0.45 μm filter membrane is used for filtration.

[0043] Specifically, the solvent is selected from one or more of dichloromethane and methanol; preferably, it is dichloromethane.

[0044] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (80-100 mg): (6-70 ml); preferably, it is 80 mg (thiophene pyrimidine derivative methanesulfonate): 70 ml (dichloromethane).

[0045] Method 2: Slurry Method: A certain amount of solvent was added to the thiophene pyrimidine derivative methanesulfonate sample to obtain two suspension samples for each solvent system (the dichloromethane system was only placed at room temperature). The samples were then placed at room temperature and high temperature (50°C) for slurry preparation. The sample vial for the room temperature system (wrapped in aluminum foil to protect it from light) was placed on a Labquaker rotator and rotated 360°. The sample for the high temperature system was placed in a 50°C, 150rpm constant temperature shaker for slurry preparation. Part of the suspension sample was taken out at 1 week, 2 weeks and 3 weeks, centrifuged, and the solid residue was collected. The solvent was evaporated in a 40°C vacuum drying oven to obtain the sample, which is the crystal form I of the thiophene pyrimidine derivative methanesulfonate.

[0046] Specifically, the solvent is selected from one or more of N,N-dimethylacetamide, methyl tert-butyl ether, acetonitrile, ethanol, acetone, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, n-heptane, 1,4-dioxane, 2-butanone, isopropanol, and dichloromethane; preferably, it is acetone, ethyl acetate, and dichloromethane.

[0047] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (80-100 mg): (4-5 ml); preferably, it is 80 mg: 4 ml.

[0048] Method 3: Anti-solvent method: Weigh the thiophene pyrimidine derivative methanesulfonate sample into a glass bottle. Place the sample bottle on a magnetically heated stirrer at a water bath temperature of 35-55℃ and a stirring speed of 200-250 rpm. Add solvent sequentially, heat to promote sample dissolution, and maintain the temperature for 10-20 minutes. While hot, filter each solution through a 0.45-0.5 μm filter membrane. Transfer the filtrate to a new sample bottle. Slowly add the anti-solvent dropwise to the glass bottle while magnetically stirring. Centrifuge the solvent system and collect the solid. Evaporate the solvent at room temperature. The obtained sample is crystal form I of the thiophene pyrimidine derivative methanesulfonate. Allow the solvent system without solid precipitation to cool naturally to room temperature and stir magnetically. If no precipitation occurs, stir openly until solid precipitates. The obtained sample is crystal form I of the thiophene pyrimidine derivative methanesulfonate. Preferably, the water bath temperature is about 50°C, the rotation speed is 200 rpm, an appropriate amount of solvent is added, the sample is heated to promote dissolution, the temperature is maintained for 15 minutes, and the solution is filtered through a 0.45 μm filter membrane while it is still hot.

[0049] Specifically, the solvent is selected from one or more of dichloromethane, pure water, dimethyl sulfoxide, methanol, and N,N-dimethylacetamide;

[0050] The antisolvent is selected from one or more of the following: tetrahydrofuran, isopropanol, 1,4-dioxane, acetone, tetrahydrofuran, dichloromethane, ethyl acetate, isopropanol, acetonitrile, 2-butanone, ethyl acetate, isopropanol, isopropyl ether, ethyl acetate, tetrahydrofuran, etc.

[0051] Preferably, the solvent (and antisolvent) is selected from one or more of dichloromethane (tetrahydrofuran), pure water (isopropanol, 1,4-dioxane), dimethyl sulfoxide (acetone, tetrahydrofuran, dichloromethane, ethyl acetate, isopropanol), methanol (acetonitrile, 2-butanone, ethyl acetate, isopropanol, isopropyl ether), and N,N-dimethylacetamide (ethyl acetate, tetrahydrofuran); more preferably, it is methanol (ethyl acetate) and N,N-dimethylacetamide (ethyl acetate);

[0052] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent and antisolvent is (80-100 mg): (1-20 ml): (5-280 ml); preferably, it is 80 mg (thiophene pyrimidine derivative methanesulfonate): 2 ml (methanol): 14 ml (ethyl acetate) or 80 mg (thiophene pyrimidine derivative methanesulfonate): 1 ml (N,N-dimethylacetamide): 7 ml (ethyl acetate).

[0053] Method 4: Heating-Rapid Cooling (HFC) Method: Weigh an appropriate amount of thiophene pyrimidine derivative methanesulfonate sample into a suitable volume glass bottle. Place the sample bottle on a magnetically heated stirrer at a water bath temperature of approximately 45-55℃ and a stirring speed of 200-250 rpm. Add an appropriate amount of solvent, heat to promote sample dissolution, and maintain the temperature for 10-20 minutes. While hot, filter each solution through a 0.45-0.5 μm filter membrane. Transfer the filtrate to a new sample bottle and immediately place the bottle in a -20℃ freezer. After centrifuging the solvent system that precipitates a large amount of solid, remove the solid and evaporate the solvent at room temperature to obtain the sample, which is crystal form I of the thiophene pyrimidine derivative methanesulfonate. If very little or no solid precipitates, remove the sample, protect it from light at room temperature, and magnetically stir until solid precipitates. Then, centrifuge and collect the precipitated solid to obtain the sample, which is crystal form I of the thiophene pyrimidine derivative methanesulfonate. Preferably, the water bath temperature is about 50°C, the rotation speed is 200 rpm, an appropriate amount of solvent is added, the sample is heated to promote dissolution, the temperature is maintained for 15 minutes, and the solution is filtered through a 0.45 μm filter membrane while it is still hot.

[0054] Specifically, the solvent is selected from one or more of dimethyl sulfoxide, methanol, and acetonitrile; preferably, it is methanol.

[0055] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (50-100 mg): (0.9-30 ml); preferably, it is 100 mg (thiophene pyrimidine derivative methanesulfonate): 2 ml (methanol).

[0056] Method 5: Solution Heating-Slow Cooling Method (HSC): Weigh an appropriate amount of thiophene pyrimidine derivative methanesulfonate sample into a suitable volume glass bottle. Place the sample bottle on a magnetically heated stirrer at a water bath temperature of approximately 45-55℃ and a stirring speed of 200-250 rpm. Add an appropriate amount of solvent, heat to promote sample dissolution, and maintain the temperature for 15 minutes. While still hot, filter each solution through a 0.45-0.5 μm filter membrane. Transfer the filtrate to a new sample bottle and slowly cool it to room temperature at a rate of 6℃ / h. Centrifuge the solvent system from which a large amount of solid precipitates, remove the solid, and evaporate the solvent at room temperature to obtain the sample as crystal form I of the thiophene pyrimidine derivative methanesulfonate. If very little or no solid precipitates, remove the sample, protect it from light at room temperature, and magnetically stir until solid precipitates. Then, centrifuge and collect the precipitated solid to obtain the sample as crystal form I of the thiophene pyrimidine derivative methanesulfonate. Preferably, the water bath temperature is about 50°C, the rotation speed is 200 rpm, an appropriate amount of solvent is added, the sample is heated to promote dissolution, the temperature is maintained for 15 minutes, and the solution is filtered through a 0.45 μm filter membrane while it is still hot.

[0057] Specifically, the solvent is selected from one or more of methanol and ethanol; preferably, it is methanol.

[0058] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (50-60 mg): (1-10 ml); preferably, it is 50 mg (thiophene pyrimidine derivative methanesulfonate): 1 ml (methanol).

[0059] The present invention also provides a method for preparing crystal form II of the thiophene pyrimidine derivative methanesulfonate as described above, the preparation method comprising the following steps:

[0060] Place the thiophene pyrimidine derivative methanesulfonate sample in a glass bottle of appropriate volume. Place the sample bottle on a magnetically heated stirrer at a water bath temperature of approximately 35-55℃ and a stirring speed of 200-250 rpm. Add an appropriate amount of solvent and heat to promote sample dissolution. Maintain the temperature for 10-20 minutes. While still hot, filter the solution through a 0.45-0.5 μm filter membrane. Transfer the filtrate to a new sample bottle. Slowly add the antisolvent dropwise to the glass bottle while magnetically stirring. Centrifuge the solvent system from which solids precipitate and collect the solids. Evaporate the solvent at room temperature. The resulting sample is crystal form II of the thiophene pyrimidine derivative methanesulfonate. Allow the solvent system without solid precipitation to cool naturally to room temperature and then stir magnetically. If no solids precipitate, stir openly until solids precipitate. The resulting sample is crystal form II.

[0061] Preferably, the water bath temperature is about 40°C, the rotation speed is 200 rpm, an appropriate amount of solvent is added, the sample is heated to promote dissolution, the temperature is maintained for 15 minutes, and the solution is filtered through a 0.45 μm filter membrane while it is still hot.

[0062] Specifically, the solvent is selected from one or more of dichloromethane, etc.; the antisolvent is selected from one or more of ethyl acetate, etc.; preferably, the solvent is dichloromethane, and the antisolvent is ethyl acetate;

[0063] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate, the solvent, and the antisolvent is 80 mg: 70 ml: 490 ml; preferably, it is 80 mg: 70 ml: 490 ml.

[0064] Specifically, the stirring time is 10-20 minutes, preferably 15 minutes.

[0065] The present invention also provides a method for preparing crystal form III of the thiophene pyrimidine derivative methanesulfonate as described above, the preparation method comprising the following steps:

[0066] Place the thiophene pyrimidine derivative methanesulfonate sample in a glass vial of appropriate volume. Place the vial on a magnetically heated stirrer at approximately 45-55°C and 200-250 rpm. Add an appropriate amount of solvent and heat to promote sample dissolution. Maintain this temperature for 10-20 minutes. While still hot, filter the solution through a 0.45-0.5 μm filter membrane. Transfer the filtrate to a new vial and immediately place the vial in a -20°C freezer. Centrifuge the solvent system from which a large amount of solid precipitates, and collect the solid. Evaporate the solvent at room temperature to obtain crystal form III of the thiophene pyrimidine derivative methanesulfonate. If very little or no solid precipitates, remove the sample and, in the dark at room temperature, magnetically stir until solid precipitates. Collect the precipitated solid by centrifugation to obtain crystal form III. Preferably, the water bath temperature is about 50°C, the rotation speed is 200 rpm, an appropriate amount of solvent is added, heating promotes sample dissolution, and the solution is kept warm for 15 minutes. While still hot, the solution is filtered through a 0.45 μm filter membrane, and the filtrate is transferred to a new sample vial. The vial is then immediately placed in a -20°C refrigerator.

[0067] Specifically, the solvent is selected from one or more of n-butanol, etc.; preferably, it is n-butanol.

[0068] Specifically, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is 50 mg: 40 ml; preferably, it is 50 mg: 40 ml (n-butanol).

[0069] Specifically, the stirring time is 10-20 minutes, preferably 15 minutes.

[0070] The "crystal form of thiophene pyrimidine derivative methanesulfonate" proposed in this invention is the first of its kind proposed in this invention.

[0071] The present invention also provides a pharmaceutical / pharmaceutical composition comprising the crystal form of the thiophene pyrimidine derivative mesylate as described above.

[0072] Furthermore, the drug / drug composition comprises a pharmaceutically acceptable carrier.

[0073] Preferably, the pharmaceutically acceptable carrier refers to a carrier that, when properly administered to animals or humans, does not produce adverse, allergic, or other adverse reactions. Pharmaceutically acceptable carriers include, but are not limited to: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and methylcellulose; tragacanth gum powder; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers such as Tween; wetting agents such as sodium lauryl sulfate; colorants; flavoring agents; tableting agents; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic salt solutions; and phosphate buffers, etc. These substances are used as needed to help stabilize the formulation or to improve its activity or bioavailability or to produce an acceptable taste or smell when taken orally.

[0074] Preferably, the drug / drug composition may further contain physiologically compatible excipients, including buffers, diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, etc.

[0075] Specifically, the drug / drug composition is administered orally, by injection, nasal administration, transdermal administration, or mucosal administration.

[0076] Preferably, the dosage form of the drug / drug composition includes injections, sterile powders for injection, tablets, pills, capsules, lozenges, liniments, powders, granules, syrups, solutions, tinctures, aerosols, powder inhalers, or suppositories, etc. All of the above dosage forms of the drug / drug composition can be prepared according to conventional methods in the pharmaceutical field.

[0077] Specifically, the drug / drug composition includes liquid dosage forms, gaseous dosage forms, solid dosage forms, and semi-solid dosage forms.

[0078] Preferably, the route of administration of the drug / drug composition is parenteral, injection, or oral. Injection preferably includes intravenous, intramuscular, intraperitoneal, intradermal, or subcutaneous injection. The drug / drug composition is in various dosage forms conventional in the art, preferably in solid, semi-solid, gaseous, or liquid form, i.e., aqueous, non-aqueous, or suspension, more preferably tablets, capsules, granules, injections, or infusions. More preferably, it is administered intravascularly, subcutaneously, intraperitoneally, or intramuscularly. Preferably, the drug / drug composition can also be administered as an aerosol or coarse spray, i.e., nasal administration; or intrathecal, intramedullary, or intraventricular administration. More preferably, the drug / drug composition can also be administered transdermally, percutaneously, topically, intraenterically, intravaginally, sublingually, or rectally. The drug / drug composition of the present invention can be formulated into various dosage forms as needed, and the physician can determine the beneficial dosage for the patient based on factors such as patient type, age, weight, general disease condition, and administration method. Administration methods may include injection or other treatment methods.

[0079] The dosage level of the drug / drug composition of the present invention can be adjusted according to the amount of composition required to achieve the desired diagnostic or therapeutic outcome. The administration regimen can also be a single injection or multiple injections, or adjustments thereof. The selected dosage level and regimen are rationally adjusted based on various factors including the activity and stability (i.e., half-life) of the cellular drug / drug composition, the formulation, the route of administration, combination with other drugs or treatments, the disease or condition to be detected and / or treated, and the health status and prior medical history of the subject to be treated.

[0080] The therapeutically effective dose of the drug / pharmaceutical composition of the present invention can initially be estimated in cell culture experiments or animal models such as rodents, rabbits, dogs, pigs, and / or primates. Animal models can also be used to determine suitable concentration ranges and routes of administration. These can then be used to determine the useful dose and route of administration in humans. Generally, the determination and adjustment of the effective amount or dose, and the assessment of when and how to make such adjustments, are known to those skilled in the art.

[0081] For further guidance on formulations, dosages, administration regimens, and measurable treatment outcomes, see Berkow et al. (2000) The Merck Manual of Medical Information and Merck & Co. Inc., Whitehouse Station, New Jersey; Ebadi (1998) CRC Desk Reference of Clinical Pharmacology, etc.

[0082] Furthermore, the drug / drug composition may be used alone and / or in combination with one or more drugs that regulate protein kinases involved in multiple disease states, antitumor drugs that inhibit one or more biological targets, chemotherapy drugs, radiotherapy, immunotherapy drugs, etc.

[0083] The term "combined use" includes the co-administration of the compounds of the present invention with other biologically active ingredients (e.g., but not limited to, a second different antitumor drug) and non-pharmacological therapies (e.g., but not limited to, surgery or radiation therapy). For example, the compounds of the present invention can be used with other pharmaceutically active compounds, preferably compounds capable of enhancing the effects of the compounds of the present invention. The compounds of the present invention can be administered simultaneously (as a single formulation or separate formulation) or sequentially with other drug therapies. Generally, combination therapy involves the administration of two or more drugs in a single cycle or course of therapy.

[0084] Specifically, the pharmaceutical / pharmaceutical composition described in this invention can be administered co-administered with one or more other pharmaceuticals that regulate protein kinases involved in various disease states. Examples of these kinases may include, but are not limited to, serine / threonine-specific kinases, receptor tyrosine-specific kinases, and non-receptor tyrosine-specific kinases. Serine / threonine kinases include mitosis-activated protein kinase (MAPK), meiosis-specific kinase (Aurora), RAF, and Aurora kinase. Examples of receptor kinase families include epidermal growth factor receptor (EGFR) (e.g., HER2 / neu, HER3, HER4, ErbB, ErbB2, ErbB3, ErbB4, Xmrk, DER, Let23); fibroblast growth factor (FGF) receptors (e.g., FGF-R1, GFF-R2 / BEK / CEK3, FGF-R3 / CEK2, FGF-R4 / TKF, KGF-R); hepatocyte growth / diffusion factor receptor (HGFR) (e.g., MET, RON, SEA, SEX); and insulin receptors. (e.g., IGFI-R); Eph (e.g., CEK5, CEK8, EBK, ECK, EEK, EHK-1, EHK-2, ELK, EPH, ERK, HEK, MDK2, MDK5, SEK); Axl (e.g., Mer / Nyk, Rse); RET; and platelet-derived growth factor receptor (PDGFR) (e.g., PDGFα-R, PDGβ-R, CSF1-R / FMS, SCF-R / C-KIT, VEGF-R / FLT, NEK / FLK1, FLT3 / FLK2 / STK-1). Non-receptor tyrosine kinase families include, but are not limited to, BCR-ABL (e.g., p43abl, ARG); BTK (e.g., ITK / EMT, TEC); CSK, FAK, FPS, JAK, SRC, BMX, FER, CDK, and SYK.

[0085] Specifically, the drug / drug composition described in this invention can be combined with antitumor drugs (e.g., small molecules, monoclonal antibodies, antisense RNA, and fusion proteins) that inhibit one or more biological targets, such as vorinostat (Zolinza), tykerb, Gleevec, Sutent, Sprycel, Nexavar, Sorafinib, CNF2024, RG108, BMS387032, Affinitak, Avastin, Herceptin, Erbitux, AG24322, PD325901, ZD6474, PD184322, Obatadax, ABT737, and AEE788. Such combinations can enhance efficacy compared to the effects achieved by any single drug and can prevent or delay the emergence of drug-resistant mutants.

[0086] Specifically, the drug / pharmaceutical composition described in this invention is administered in combination with a chemotherapy drug. The chemotherapy drug is administered at various stages of the disease to shrink tumors, destroy cancer cells that remain after surgery, and cause, maintain, and / or alleviate symptoms associated with cancer or its treatment. Examples of these drugs include, but are not limited to, alkylated drugs such as mustard gas derivatives (nitrogen mustard, cyclophosphamide, chlorambucil, melphalan, ifosfamide), ethylenimine (thiotepa, hexamethylmelamine), alkyl sulfonates / esters (busulfan), hydrazine and triazine (hexamethylmelamine, procarbazine, dazometamide and temozolomide), nitrosoureas (carmustine, lomustine and streptozotocin), ifosfamide and metal salts (carboplatin, cisplatin, and oxaliplatin); plant alkaloids such as podophyllotoxin (etoposide and tenisopide), taxanes (paclitaxel and docetaxel), vinca alkaloids (vincristine, vincristine, vinorelbine and vinorelbine), and camptothecin analogues (irinotecan and topotecan); and antitumor antibiotics such as chromomycin (actinomycin D and procamycin). Anthracyclines (doxorubicin, daunorubicin, epirubicin, mitoxantrone, pentorubicin, and demethoxydaunorubicin), and other antibiotics such as mitomycin, actinomycin D, and bleomycin; antimetabolites such as folic acid antagonists (methotrexate, pemetrexed, raltitrexed, aminopterin), pyrimidine antagonists (5-fluorouracil, fluorouracil deoxyribonucleoside, cytarabine, capecitabine, and gemcitabine), and purine antagonists (6-mercaptocyanide, thiouracil, thiouracil, thiouracil, and gemcitabine). Purine and 6-thioguanine) and adenosine deaminase inhibitors (cladribine, fludarabine, mercaptopurine, clofarabine, thioguanine, nerabine, and pentostatin); topoisomerase inhibitors such as topoisomerase I inhibitors (irinotecan, topotecan) and topoisomerase II inhibitors (acridine, etoposide, etoposide phosphate, teniposide); monoclonal antibodies (alendumab, gemtuzumab) (ozogamicin), rituximab, trastuzumab, tiemomumab, cetuximab, panitumumab, tosimomumab, bevacizumab); and other antitumor drugs such as ribonucleotide reductase inhibitors (hydroxyurea); corticosteroid inhibitors (mitotan); enzymes (asparaginase and pegaspargase); antimicrotubule drugs (estradiol mustard); and retinoids (bexarotin, isotretinoin, retinoic acid (ATRA)).

[0087] Specifically, the drug / drug composition described in this invention is administered together with a chemotherapy drug. The chemotherapy drug serves to protect the body or minimize the side effects of chemotherapy. Examples of such drugs include, but are not limited to, Amfostine, mesna, and dexrazoxane.

[0088] In one aspect of the invention, the drug / drug composition described herein is administered in conjunction with radiotherapy. The radioactive material is typically delivered inward (to implant radioactive material near the cancer site) or outward from a machine employing proton (X-ray or gamma-ray) or particle radiation. When the combination therapy also includes radiotherapy, the radiotherapy can be administered at any suitable time, as long as the combined effect of the therapeutic drug and the radiotherapy is achieved. For example, in suitable cases, the beneficial effect can still be achieved when radiotherapy is temporarily suspended for several days or even weeks while the therapeutic drug is being administered.

[0089] Specifically, the pharmaceutical / pharmaceutical composition described in this invention can be used in combination with immunotherapeutic drugs. One form of immunotherapy is to induce an active, systemic, tumor-specific immune response in the host by administering a vaccine composition at a site distant from the tumor. Various types of vaccines have been proposed, including isolated tumor antigen vaccines and vaccines against individual genes. Another approach is to use tumor cells from the subject to be treated, or derivatives of such cells (reviewed by Schirrmacher et al. (1995) J. Cancer Res. Clin. Oncol. 121:487). In U.S. Patent 5,484,596, Hanna Jr. et al. claim protection for a method of treating resectable cancer to prevent recurrence or metastasis, comprising surgically removing the tumor, dispersing the cells with collagenase, irradiating the cells, and immunizing the patient with at least three consecutive doses of approximately 10⁷ cells.

[0090] In one embodiment, the compounds of the present invention can be used to induce or inhibit apoptosis, a physiological cell death process crucial for normal development and homeostasis. Alterations in the apoptosis pathway contribute to the pathogenesis of a variety of human diseases. The compounds of the present invention, as regulators of apoptosis, can be used to treat a variety of human diseases associated with apoptosis abnormalities, including cancers (particularly, but not limited to, follicular lymphoma, cancers associated with p53 mutations, hormone-related tumors of the breast, prostate, and ovaries, and precancerous lesions such as familial adenomatous polyposis), viral infections (including, but not limited to, herpesviruses, poxviruses, Epstein-Barr virus, Sindbis virus, and adenoviruses), autoimmune diseases (including, but not limited to, lupus erythematosus, systemic lupus erythematosus, immune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes), blood disorders (chronic anemia and aplastic anemia), and cancer pain.

[0091] The pharmaceutical compositions described in this invention may be included in a container, package, or dispenser along with instructions for administration.

[0092] The preparation of pharmaceutical compositions comprising the active ingredient of this invention is well known in the art, for example, through mixing, granulation, or tableting processes. The active therapeutic ingredient is typically mixed with a pharmaceutically acceptable excipient compatible with the active ingredient. For oral administration, the active agent is mixed with additives commonly used for this purpose, such as carriers, stabilizers, or inert diluents, and converted by common methods into a suitable form of administration, such as tablets, coated tablets, hard or soft gel capsules, aqueous solutions, alcoholic solutions, or oil solutions, as described above.

[0093] The present invention also provides the crystal form of the thiophene pyrimidine derivative mesylate salt as described above, or the preparation method as described above, or the use of the drug / drug composition as described above in the preparation of a drug for treating diseases caused by abnormal cell proliferation due to overexpression of EGFR.

[0094] The beneficial effects of this invention include: Through the research and development of thiophene pyrimidine derivatives protected by patents (US10829495B2 / CN108289895B), the present invention has successfully screened out the crystal forms of stable thiophene pyrimidine derivative mesylate salts for the first time, and these crystals have been successfully used in preclinical and clinical studies. Attached Figure Description

[0095] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0096] Figure 1 This is an XRPD overlay of solid samples precipitated from various solvent systems (water, dichloromethane, methanol) in the solution evaporation method of Example 1 of the present invention; where intensity represents intensity and counts represents counts.

[0097] Figure 2 This is an XRPD overlay image of samples obtained at 1 week, 2 weeks and 3 weeks in the suspension equilibrium method of Example 1 of this invention (n-heptane-room temperature, tetrahydrofuran-room temperature, 1,4-dioxane-room temperature, 1,4-dioxane-50°C); where slurry represents the feed solution; W represents the week; and RT represents the room temperature.

[0098] Figure 3 This is XRPD overlay images of samples obtained at 1 week, 2 weeks, and 3 weeks in the suspension equilibrium method of Example 1 of this invention (tetrahydrofuran-room temperature, tetrahydrofuran-50°C, ethyl acetate-room temperature, ethyl acetate-50°C); where intensity represents intensity and counts represents counts.

[0099] Figure 4 This is XRPD overlay images of samples obtained at 1 week, 2 weeks, and 3 weeks in the suspension equilibrium method of Example 1 of this invention (acetone-room temperature, acetone-50°C, ethanol-room temperature, ethanol-50°C); where intensity represents intensity and counts represents counts.

[0100] Figure 5 This is XRPD overlay image-4 of samples obtained at 1 week, 2 weeks and 3 weeks in the suspension equilibrium method of Example 1 of the present invention for each solvent system (acetonitrile-room temperature, acetonitrile-50℃, 2-butanone-room temperature, 2-butanone-50℃); where intensity represents intensity and counts represents counts.

[0101] Figure 6 This is XRPD overlay images of samples obtained at 1 week, 2 weeks, and 3 weeks in the suspension equilibrium method of Example 1 of this invention (dichloromethane-room temperature, methyl tert-butyl ether-room temperature, N,N-dimethylacetamide-room temperature); where intensity represents intensity and counts represents counts.

[0102] Figure 7 This is an XRPD overlay of the solid sample precipitated by the antisolvent method (good solvent: dichloromethane) in Example 1 of the present invention; where intensity represents intensity; counts represents count; and anti represents antisolvent.

[0103] Figure 8 This is an XRPD overlay image of the solid sample precipitated by the antisolvent method (good solvent: pure water) in Example 1 of the present invention; where intensity represents intensity; counts represents count; and anti represents antisolvent.

[0104] Figure 9 This is an XRPD overlay image of the solid sample precipitated by the antisolvent method (good solvent: dimethyl sulfoxide) in Example 1 of this invention;

[0105] Figure 10 This is an XRPD overlay image of the solid sample precipitated by the antisolvent method (good solvent: methanol) in Example 1 of this invention;

[0106] Figure 11 This is an XRPD overlay image of the solid sample precipitated by the antisolvent method (good solvent: N,N-dimethylacetamide) in Example 1 of the present invention;

[0107] Figure 12 This is an XRPD overlay of solid samples precipitated in each solvent system using the solution heating-rapid cooling method in Example 1 of this invention;

[0108] Figure 13This is an XRPD overlay of solid samples precipitated in each solvent system using the solution heating-slow cooling method in Example 1 of this invention;

[0109] Figure 14 This is an XRPD overlay of the polymorphs (crystal forms I to III) of the thiophene pyrimidine derivative mesylate in Example 1 of this invention;

[0110] Figure 15 This is a polarized light microscope image (×100) of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0111] Figure 16 This is the XRPD image of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0112] Figure 17 This is an XRPD image (with numerical labels) of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0113] Figure 18 This is the XRPD peak finding report of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention;

[0114] Figure 19 This is the DSC & TGA diagram of crystal form I of the thiophene pyrimidine derivative mesylate in Example 1 of this invention;

[0115] Figure 20 It is crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention. 1 H NMR spectrum;

[0116] Figure 21 This is a DSC image of the sample of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention after heating to 150°C;

[0117] Figure 22 This is an XRPD overlay image of crystal form I of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention before and after heating;

[0118] Figure 23 This is the XRPD image of crystal form II of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention;

[0119] Figure 24 This is an XRPD image (with numbers) of crystal form II of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0120] Figure 25 This is the XRPD peak finding report of crystal form II of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention;

[0121] Figure 26This is the DSC & TGA diagram of crystal form II of the thiophene pyrimidine derivative mesylate in Example 1 of this invention;

[0122] Figure 27 It is crystal form II of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention. 1 H NMR spectrum;

[0123] Figure 28 This is the XRPD image of crystal form II of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention after heating to 150°C;

[0124] Figure 29 This is the XRPD image of crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention;

[0125] Figure 30 This is an XRPD image (with numbers) of crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0126] Figure 31 This is the XRPD peak finding report of crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention;

[0127] Figure 32 This is the DSC & TGA diagram of crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention;

[0128] Figure 33 It is the crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of this invention. 1 HNMR spectrum;

[0129] Figure 34 This is the XRPD image of crystal form III of the thiophene pyrimidine derivative methanesulfonate in Example 1 of the present invention after heating to 150°C.

[0130] In the accompanying drawings of this invention, intensity represents strength; counts represents counting; slurry represents liquid; W represents week; RT represents room temperature; Anti represents anti-solvent; HeatFlow represents heat flow; Temperature represents temperature; WeightChange represents weight change; WeightLoss represents weight loss; WeightPercentLoss represents weight loss percentage; and Normalized represents standardization. Detailed Implementation

[0131] The present invention will be further described in detail below with reference to specific embodiments. Except for the contents specifically mentioned below, the processes, conditions, and experimental methods for implementing the present invention are all common knowledge and general knowledge in the art, and the present invention does not have any particular limitations.

[0132] The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0133] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values.

[0134] This invention discloses a crystal form of a thiophene pyrimidine derivative methanesulfonate, its preparation method, and its application. The thiophene pyrimidine derivative methanesulfonate comprises crystal form I, crystal form II, and crystal form III. Crystal form I is a single-melting-point anhydrous crystal form with characteristic XRPD diffraction peaks; its initial melting point is around 226°C; and it exhibits good physical and chemical stability within two weeks under high temperature (60°C) and accelerated conditions (40°C / 75% RH). This invention also discloses the application of the crystal form of the thiophene pyrimidine derivative methanesulfonate and its preparation method in the preparation of a drug for treating diseases caused by abnormal cell proliferation due to EGFR overexpression. This invention has broad application prospects.

[0135] Unless otherwise specified, the experimental materials used in the examples are all conventional biochemical reagents.

[0136] Example 1: Crystal form screening experiment of thiophene pyrimidine derivative methanesulfonate

[0137] 1. Approximate solubility (to provide available solvents for screening)

[0138] In this invention, approximately 4 mg of thiophene pyrimidine derivative methanesulfonate was weighed into a 4 mL glass bottle. Solvent was added gradually in small amounts at room temperature and 50°C, and the sample was sonicated while observing the dissolution. Solvent addition was stopped when the solvent volume reached 4 mL. The approximate solubility values ​​of thiophene pyrimidine derivative methanesulfonate in each solvent were calculated at room temperature and 50°C.

[0139] Solubility results (see Table 1) show that at room temperature, the thiophene pyrimidine derivative methanesulfonate compounds are slightly soluble in methanol and dimethyl sulfoxide, sparingly soluble in pure water, N,N-dimethylacetamide, and dichloromethane, and poorly soluble in the other solvents listed in Table 1. At 50°C, the thiophene pyrimidine derivative methanesulfonate compounds are readily soluble in pure water and dimethyl sulfoxide, soluble in methanol, slightly soluble in N,N-dimethylacetamide, sparingly soluble in ethanol, and poorly soluble in the other solvents listed in Table 1. At 40°C, the thiophene pyrimidine derivative methanesulfonate compounds are slightly soluble in dichloromethane (dichloromethane has a boiling point of 39.8°C).

[0140] Table 1. Approximate solubility of thiophenepyrimidine derivative methanesulfonates in different solvents

[0141]

[0142]

[0143] Note: In Table 1, NA indicates none or blank.

[0144] 2. Preparation of crystal forms of thiophene pyrimidine derivative methanesulfonate in the screening experiment of this invention.

[0145] The experimental methods for preparing the crystal forms of the thiophene pyrimidine derivative methanesulfonate of this invention include: solution evaporation crystallization, suspension equilibrium method, heating-rapid / slow cooling crystallization method, and antisolvent method. Three new crystal forms were discovered, yielding crystal form I (amorphous), crystal form II (1 molecule ethyl acetate solvate), and crystal form III (1 molecule n-butanol solvate), respectively. Crystal form I is the crystal form of the thiophene pyrimidine derivative methanesulfonate described in this invention. The experimental results for preparing the above crystal forms are shown in Table 2.

[0146] Table 2. Summary of experimental results for the preparation of crystal forms of thiophene pyrimidine derivative mesylates.

[0147] Experimental methods Number of experiments result Solution evaporation crystallization (EVA) 3 Crystal form I Room temperature suspension equilibrium method (slurry) 11 Crystal form I 50℃ Suspension Equilibrium Method (Pulping) (Slurry-50℃) 8 Crystal form I Anti-solvent method 23 Crystal form I, Crystal form II Heating-and-cooling crystallization (HFC) method 5 Crystal form I, Crystal form III Solution heating-slow cooling crystallization (HSC) 7 Crystal form I total 57 Crystal form I, Crystal form II, Crystal form III

[0148] 2.1 Crystal form screening - solution evaporation method (EVA)

[0149] In this invention, three portions of thiophene pyrimidine derivative methanesulfonate samples, approximately 80 mg / portion, were weighed and placed in glass bottles. An appropriate amount of the solvent corresponding to Table 2 was added, and the samples were sonicated to promote dissolution. The samples were then filtered through a 0.45 μm filter membrane into new sample bottles. The sample bottles were placed open in a fume hood to allow the solvent to evaporate naturally at room temperature. After a large amount of solid precipitated, the samples were characterized by XRPD.

[0150] Experimental results showed that the sample obtained in the dichloromethane and methanol system was crystalline form I; the sample obtained in the aqueous system was amorphous. The experimental results of the slow evaporation method are summarized in Table 3 and... Figure 1 .

[0151] Table 3 Summary of experimental results using the solution evaporation method

[0152] Serial Number solvent Feeding amount (mg) Volume (ml) Sample appearance Crystal form 1 dichloromethane 80 70 Yellow solid Crystal form I 2 methanol 80 6 Yellow solid Crystal form I 3 water 80 35 Yellow transparent solid amorphous

[0153] 2.2 Crystal form screening—Slurry method

[0154] In this invention, 19 appropriate amounts of thiophene pyrimidine derivative methanesulfonate samples were weighed and added sequentially with certain amounts of solvent (see Tables 4 and 5) to obtain two suspension samples for each solvent system (the dichloromethane system was only placed at room temperature). The samples were then placed at room temperature and at a high temperature (50°C) for slurry preparation. The sample vials of the room temperature system (wrapped in aluminum foil to protect from light) were placed on a Labquaker rotator and rotated 360°. The samples of the high temperature system were placed in a 50°C, 150rpm constant temperature shaker for slurry preparation. At 1, 2, and 3 weeks, a portion of the suspension samples was taken out, centrifuged, and the solid residue was collected. The solvent was evaporated in a 40°C vacuum drying oven, and then characterized by XRPD.

[0155] The screening results of the suspension equilibrium method are summarized in Tables 4 and 5. XRPD results are shown below. Figures 2-6 The results showed that after equilibration for 1, 2, and 3 weeks (at room temperature and high temperature) in N,N-dimethylacetamide, methyl tert-butyl ether, acetonitrile, ethanol, acetone, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, n-heptane, 1,4-dioxane, 2-butanone, isopropanol, and dichloromethane, crystal form I was obtained, and the crystal form remained unchanged.

[0156] Table 4. Summary of experimental results of the suspension equilibrium method (room temperature system)

[0157]

[0158]

[0159] Table 5. Summary of experimental results of the suspension equilibrium method (high temperature 50℃ system)

[0160]

[0161] 2.3 Crystal form screening - Anti-solvent method

[0162] In this invention, approximately 5 portions of thiophene pyrimidine derivative methanesulfonate samples were weighed into glass bottles of suitable volume. The sample bottles were placed on a magnetically heated stirrer at a water bath temperature of approximately 50°C (or a dichloromethane water bath temperature of approximately 40°C) and a stirring speed of 200 rpm. Appropriate amounts of the solvents listed in Table 7 were added sequentially, and the mixture was heated to promote sample dissolution. The mixture was kept at this temperature for 15 minutes. While still hot, each solution was filtered through a 0.45 μm filter membrane. The filtrate was then transferred to a new sample bottle and divided into equal portions. Different antisolvents were slowly added dropwise to each glass bottle under magnetic stirring. The solvent system from which solids precipitated was centrifuged and the solids were collected. The solvent was evaporated at room temperature and characterized by XRPD. The solvent system from which no solids precipitated was allowed to cool naturally to room temperature and then magnetically stirred. If no solids precipitated, the mixture was stirred open until solids precipitated.

[0163] Antisolvent screening results (Table 6 and Figures 7-11The results indicate that in screening experiments using pure water, dimethyl sulfoxide, methanol, and N,N-dimethylacetamide as good solvents, crystal form I or amorphous forms were obtained. In the dichloromethane system, no solid precipitated when using acetone and isopropanol as antisolvents; the sample obtained in the experiment using tetrahydrofuran as an antisolvent was crystal form I of the thiophene pyrimidine derivative methanesulfonate; and the sample obtained in the screening experiment using ethyl acetate as an antisolvent was crystal form II of the thiophene pyrimidine derivative methanesulfonate.

[0164] Table 6 Summary of experimental results of the anti-solvent method

[0165]

[0166]

[0167] 2.4 Crystal form screening-solution heating-rapid cooling method (HFC)

[0168] In this invention, five portions of thiophene pyrimidine derivative methanesulfonate samples, each approximately 50–200 mg, are weighed into glass vials of suitable volume. The vials are placed on a magnetically heated stirrer at approximately 50°C and 200 rpm. Appropriate amounts of the solvents listed in Table 8 are added sequentially, and heating is used to promote sample dissolution. The mixture is kept at this temperature for 15 minutes. While still hot, each solution is filtered through a 0.45 μm filter membrane. The filtrate is then transferred to a new vial, and the vial is immediately placed in a -20°C freezer. For solvent systems that precipitate a large amount of solid, the solid is centrifuged, and the solvent is evaporated at room temperature. The solid is then characterized using XRPD. If very little or no solid precipitates, the sample is removed, kept at room temperature in the dark, and magnetically stirred until solid precipitates. The precipitated solid is then collected by centrifugation and characterized using XRPD.

[0169] Screening results of the solution heating-rapid cooling method (see Table 7 and ) Figure 12 The results showed that the samples obtained in the dimethyl sulfoxide, methanol and acetonitrile system were all of crystal form I; a new crystal form III was obtained in the n-butanol system, and the sample obtained in the water system was amorphous.

[0170] Table 7 Summary of experimental results for the solution heating-rapid cooling method

[0171] Serial Number solvent Feeding amount (mg) Volume (ml) Sample appearance Crystal form 1 dimethyl sulfoxide 100 0.9 Yellow solid Crystal form I 2 methanol 100 2.0 Yellow solid Crystal form I 3 Acetonitrile 50 30.0 Yellow solid Crystal form I 4 n-Butanol 50 40.0 Yellow solid Crystal form III 5 water 200 1.0 Yellow transparent solid amorphous

[0172] 2.5 Crystal form screening - solution heating - slow cooling method (HSC)

[0173] In this invention, seven portions of thiophene pyrimidine derivative methanesulfonate samples, each approximately 20–100 mg, are weighed into glass vials of suitable volume. The vials are placed on a magnetically heated stirrer at approximately 50°C and 200 rpm. Appropriate amounts of the solvents listed in Table 9 are added sequentially, and heating is used to promote sample dissolution. The mixture is kept at this temperature for 15 minutes. While still hot, each solution is filtered through a 0.45 μm filter membrane. The filtrate is transferred to a new vial and slowly cooled to room temperature at a rate of 6°C / h. The solvent system from which a large amount of solid precipitates is centrifuged, and the solid is collected and allowed to evaporate the solvent at room temperature. If only a very small amount or no solid precipitates, the sample is removed and magnetically stirred at room temperature in the dark until solid precipitates. The precipitated solid is then collected by centrifugation and characterized using XRPD.

[0174] Screening results of the solution heating-rapid cooling method (Table 8 and Figure 13 The results showed that no solid precipitated in the acetonitrile, N,N-dimethylacetamide, dichloromethane and acetone system; the sample obtained in the ethanol and methanol system was crystal form I of the thiophene pyrimidine derivative methanesulfonate; and the sample obtained in the aqueous system was amorphous thiophene pyrimidine derivative methanesulfonate.

[0175] Table 8 Summary of experimental results of the solution heating-slow cooling method

[0176]

[0177] 3. A method for characterizing and detecting the crystal form of thiophene pyrimidine derivative methanesulfonates prepared in the screening experiment of this invention.

[0178] The crystal forms I to III of the thiophene pyrimidine derivative methanesulfonates prepared in the crystal form screening experiment of the present invention were characterized by the following detection methods:

[0179] 3.1 Polarizing Microscopy (PLM) Observation

[0180] Take a small amount of sample, place it on a graduated glass slide, add an appropriate amount of liquid paraffin to disperse it, cover it with a coverslip, and observe the particle shape, size and crystal properties under a 10x objective lens of a microscope. Use an orthogonal polarizer to display the birefringence properties and crystal habit of the sample, and take a picture with a digital camera.

[0181] 3.2 Powder X-ray Diffraction Analysis (XRPD)

[0182] The light source was CuK, the X-ray intensity was 40KV / 40mA, the scanning mode was Theta-theta, the scanning angle range was 4° to 40°, the step size was 0.05°, and the scanning speed was 0.5 seconds / step.

[0183] 3.3 Differential Scanning Calorimetry (DSC)

[0184] Weigh an appropriate amount of sample and place it in a non-sealed aluminum dish. In a nitrogen flow environment (50 mL / min), equilibrate the sample at 25°C, and then heat it from 25°C to 300°C at a heating rate of 10°C / min.

[0185] 3.4 Thermogravimetric analysis (TGA)

[0186] Weigh an appropriate amount of sample and place it in a platinum sample pan. In an environment with nitrogen flow of sample (60 mL / min) and nitrogen flow of balance (40 mL / min), heat from room temperature to 300℃ at a heating rate of 10℃ / min.

[0187] 3.5 One-dimensional proton NMR spectrum (NMR) 1 HNMR)

[0188] Weigh approximately 5 mg of sample into an NMR tube, add 0.6 mL of deuterated dimethyl sulfoxide (DMSOd6) to dissolve the sample into a clear solution, and scan the sample solution using a Bruker Advance III 400 MHz NMR spectrometer according to the general method to acquire sample data.

[0189] 4. Analysis and conclusions of the screening experiment of this invention

[0190] The crystal form screening experiments for the thiophene pyrimidine derivative methanesulfonate of this invention were conducted using the suspension equilibrium method, the heating-rapid / slow cooling crystallization method, the antisolvent method, and the solution evaporation crystallization method. Three crystal forms were identified: crystal form I (amorphous), crystal form II (one molecule of ethyl acetate solvate), and crystal form III (one molecule of n-butanol solvate). The physicochemical properties of each crystal form are shown in Table 9.

[0191] Based on the salt form screening study, crystal form I of the thiophene pyrimidine derivative methanesulfonate is an anhydrous crystal form with a single melting point and characteristic XRPD diffraction peaks; the initial melting point is around 226℃; and it exhibits good physical and chemical stability within 2 weeks under high temperature (60℃) and accelerated conditions (40℃ / 75% RH). Therefore, this crystal form screening study recommends crystal form I of the thiophene pyrimidine derivative methanesulfonate as the superior crystal form for subsequent research and development, namely, the crystal form of the thiophene pyrimidine derivative methanesulfonate described in this invention.

[0192] Table 9 Summary of characterization results for polymorphs of thiophene pyrimidine derivative mesylates

[0193]

[0194]

[0195] Note: NA indicates none or blank.

[0196] 4.1 Polymorph screening and characterization of thiophene pyrimidine derivative mesylates

[0197] Based on the XRPD characterization of the solids obtained in the screening experiments and subsequent identification and evaluation, this invention discovered three polymorphs of the thiophene pyrimidine derivative methanesulfonate, named Form I, Form II, and Form III, respectively. The XRPD overlay images of the three polymorphs are shown below. Figure 14 .

[0198] 4.1.1 Crystal form I:

[0199] Crystal form I sample, the starting material for the crystal form screening study, is a thiophene pyrimidine derivative methanesulfonate, which appears as a yellow powder. The PLM, DSC, TGA, XRPD, and NMR characterization results of crystal form I sample are shown below. Figures 15-20 .

[0200] Under polarized light microscopy, the sample exhibits obvious birefringence and granular crystal formation with a particle size ranging from 0 to 30 μm. X-ray powder diffraction analysis shows that the sample possesses multiple distinct characteristic diffraction peaks, indicating that it is a crystalline sample. The DSC & TGA curves for crystal form I show an initial melting point of 226.05 °C. The DSC heat flow curve shows two endothermic peaks before 150 °C, corresponding to a 1.815% weight loss on the TGA curve. 1 Based on the HNMR results, it is speculated that the endothermic peak and weight loss are caused by the volatilization of a small amount of residual acetone solvent in the sample.

[0201] After heating crystal form I to 150°C using TGA, the two endothermic peaks before 150°C disappeared, indicating that the residual acetone solvent had been removed. Figure 21 Furthermore, the crystal form of the sample remained unchanged after heating, still being crystal form I. Figure 22 This indicates that acetone is only a small amount of solvent adsorbed on the surface and has not entered the crystal lattice. Based on the combined results of DSC, TGA, and heating experiments, crystal form I is an anhydrous and solvent-free crystal form (anhydrate).

[0202] 4.1.2 Crystal form II:

[0203] Crystal form II was obtained solely through antisolvent testing of crystal form I in dichloromethane-ethyl acetate; the specific preparation method is detailed in Table 6 under section 2.3. The DSC, TGA, XRPD, and NMR characterization results of the crystal form II sample are shown below. Figures 23-27 .

[0204] The crystal form II sample appears as a yellow powder. X-ray powder diffraction results show that the sample has several distinct characteristic diffraction peaks, indicating it is crystalline. DSC & TGA results show that the initial melting point of crystal form II is 225.83℃. The two endothermic peaks at 150℃ on the DSC heat flow curve correspond to a 10.45% step weight loss on the TGA, which, combined with the characteristics of crystal form II... 1Based on the HNMR results, it is inferred that the crystal form II sample is an ethyl acetate solvate (containing 1 molecule of ethyl acetate, with a theoretical content of 11.90%). Figure 28 As shown, after heating the crystal form II sample to 150°C using TGA to remove the solvent, XRPD results showed that crystal form II could be transformed into crystal form I.

[0205] 4.1.3 Crystal form III:

[0206] Crystal form III can only be obtained by rapidly cooling crystal form I in n-butanol; the specific preparation method is shown in Table 7 under section 2.4. The DSC, TGA, XRPD, and NMR characterization results of the crystal form III sample are shown below. Figures 29-33 .

[0207] The crystal form III sample appears as a yellow powder. X-ray powder diffraction results show that the sample has several distinct characteristic diffraction peaks, indicating that it is crystalline. DSC & TGA results show that the initial melting point of crystal form III is 224.78℃. The two endothermic peaks at 150℃ on the DSC heat flow curve correspond to a 10.08% step weight loss on the TGA, which, combined with the characteristics of crystal form III... 1 Based on the HNMR results, it is inferred that the crystal form III sample is a n-butanol solvate (containing 1 molecule of n-butanol, with a theoretical content of 10.20%). Figure 34 As shown, after heating the crystal form III sample to 150°C using TGA to remove the solvent, XRPD results showed that crystal form III could be transformed into crystal form I.

[0208] 4.2 Conclusion:

[0209] The crystal form screening method for the thiophene pyrimidine derivative methanesulfonate of this invention includes the suspension equilibrium method, the heating-rapid / slow cooling crystallization method, the antisolvent method, and the solution evaporation method. A total of three crystal forms were found: crystal form I (amorphous), crystal form II (1 molecule of ethyl acetate solvate), and crystal form III (1 molecule of n-butanol solvate).

[0210] Based on comprehensive crystal form screening, crystal form I is an anhydrous and solvent-free compound with a single characteristic melting point and XRPD diffraction peaks; its initial melting point is around 226℃; after being placed at high temperature (60℃) and under accelerated conditions (40℃ / 75% RH) for 2 weeks, observation and testing showed no changes in physical properties and chemical data, indicating good stability. Therefore, this crystal form screening study recommends crystal form I of the thiophene pyrimidine derivative methanesulfonate as the superior crystal form for subsequent research and development.

[0211] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0212] As used in this invention, the terms "comprising" and "including" are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0213] As used in this invention, the term "and / or" includes any one or more of the related listed items and all combinations thereof.

[0214] The scope of protection of this invention is not limited to the above embodiments. Any variations and advantages that can be conceived by those skilled in the art without departing from the spirit and scope of this invention are included in this invention and are protected by the appended claims.

Claims

1. A crystal form of a thiophene pyrimidine derivative methanesulfonate, characterized in that, The structural formula of the thiophene pyrimidine derivative methanesulfonate is shown in formula (I):

2. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 1, characterized in that, The crystal form of the thiophene pyrimidine derivative methanesulfonate is selected from the following group: crystal form I, crystal form II and crystal form III.

3. The crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form I of the thiophene pyrimidine derivative methanesulfonate shows characteristic peaks at 2θ angles of 7.379, 9.535, 10.822, 11.859, 13.377, 15.575, 17.249, 18.664, 19.353, 20.072, 21.592, 21.906, 22.402, 23.644, 25.683, and 26.

240.

4. The crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The crystal form I has one or more features selected from the group consisting of: 1) The crystal form I has a basic X-ray powder diffraction pattern as shown in Figure 17; 2) Crystal type I has a basic XRPD peak finding report as shown in Figure 18; 3) The crystal form I has a DSC spectrum as shown in Figure 19; 4) The crystal form I has a TGA spectrum as shown in Figure 19; 5) The crystal form I has the basic characteristics shown in Figure 20. 1 H NMR spectrum; 6) The crystal form I has a DSC diagram after heating to 150°C, as shown in Figure 21; 7) The crystal form I has an XRPD overlay image before and after heating, as shown in Figure 22.

5. The crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 4, characterized in that, The DSC spectrum of crystal form I includes an endothermic peak at 228.66 °C.

6. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The crystal form I is an anhydrous crystal form with a single melting point and characteristic XRPD diffraction peaks; the initial melting point is 226°C; and it exhibits good physical and chemical stability within 2 weeks under high temperature (60°C) and accelerated conditions (40°C / 75% RH).

7. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form II of the thiophene pyrimidine derivative methanesulfonate has characteristic peaks at 2θ angles of 14.001, 18.076, 18.743, 20.312, 22.602, 24.038, and 25.

686.

8. The crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The crystal form II has one or more features selected from the group consisting of: 1) The crystal form II has a basic X-ray powder diffraction pattern as shown in Figure 24; 2) The crystal form II has a basic XRPD peak finding report as shown in Figure 25; 3) The crystal form II has a DSC spectrum as shown in Figure 26; 4) The crystal form II has a TGA spectrum as shown in Figure 26; 5) The crystal form II has the basic characteristics shown in Figure 27. 1 H NMR spectrum; 6) The crystal form II has an XRPD pattern after heating to 150°C, as shown in Figure 28.

9. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 8, characterized in that, The DSC spectrum of crystal form II includes an endothermic peak at 227.05 °C.

10. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form III of the thiophene pyrimidine derivative methanesulfonate has characteristic peaks at 2θ angles of 6.005, 12.020, 13.977, 18.305, 18.722, 20.602, 22.516, 24.096, and 25.

807.

11. The crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The crystal form III has one or more characteristics selected from the group consisting of: 1) The crystal form III has a basic X-ray powder diffraction pattern as shown in Figure 30; 2) The crystal form III has a basic XRPD peak finding report as shown in Figure 31; 3) The crystal form III has a DSC spectrum as shown in Figure 32; 4) The crystal form III has a TGA spectrum as shown in Figure 32; 5) The crystal form III has the basic characteristics shown in Figure 33. 1 H NMR spectrum; 6) The crystal form III has an XRPD pattern after heating to 150°C, as shown in Figure 34.

12. The crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 11, characterized in that, The DSC spectrum of crystal form III includes an endothermic peak at 226.91 °C.

13. A method for preparing the crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The preparation method of crystal form I includes: Method 1: Solution evaporation crystallization method for EVA, comprising the following steps: placing the thiophene pyrimidine derivative methanesulfonate sample in a glass bottle, adding solvent, sonicating to promote dissolution, filtering through a 0.45-0.5 μm filter membrane into a new sample bottle, placing the sample bottle open in a fume hood, allowing the solvent to evaporate naturally at room temperature, and obtaining the sample after a large amount of solid has precipitated is the crystal form I; and / or, Method 2: Slurry suspension equilibrium method, comprising the following steps: A certain amount of solvent is added to the thiophene pyrimidine derivative methanesulfonate sample to obtain two suspension samples for each solvent system. The samples are then placed at room temperature and at a high temperature (50°C) for slurry preparation. The sample vial for the room temperature system is rotated 360° on a Labquaker rotator. The sample for the high temperature system is placed in a 50°C, 150rpm constant-temperature shaker for slurry preparation. Partial suspension samples are taken out at 1 week, 2 weeks, and 3 weeks, centrifuged, and the solid residue is collected. The solvent is evaporated in a 40°C vacuum drying oven to obtain the sample of crystal form I; and / or, Method 3: Anti-solvent method, comprising the following steps: Weigh the thiophene pyrimidine derivative methanesulfonate sample into a glass bottle, place the sample bottle on a magnetically heated stirrer, water bath temperature 35-55℃, rotation speed 200-250 rpm, add solvent sequentially, heat to promote sample dissolution, keep warm for 10-20 min, filter each solution through a 0.45-0.5 μm filter membrane while hot, transfer the filtrate to a new sample bottle, slowly add anti-solvent dropwise to the glass bottle under magnetic stirring, centrifuge the solvent system and collect the solid, evaporate the solvent at room temperature, the obtained sample is the crystal form I; the solvent system without solid precipitation is allowed to cool naturally to room temperature and magnetically stirred. If no precipitation still occurs, stir openly until solid precipitates, the obtained sample is the crystal form I; and / or, Method 4: Solution heating-rapid cooling method for HFC, comprising the following steps: Weigh the thiophene pyrimidine derivative methanesulfonate sample into a glass bottle, place the sample bottle on a magnetically heated stirrer, water bath temperature 45-55℃, rotation speed 200-250 rpm, add solvent, heat to promote sample dissolution, keep warm for 10-20 min, filter each solution through a 0.45-0.5 μm filter membrane while hot, transfer the filtrate to a new sample bottle, immediately place the sample bottle in a -20℃ freezer, centrifuge the solvent system that precipitates a large amount of solid, remove the solid, evaporate the solvent at room temperature to obtain the sample as crystal form I; if very little or no precipitation occurs, remove the sample, protect it from light at room temperature, magnetically stir until solid precipitates, then centrifuge and collect the precipitated solid to obtain the sample as crystal form I; and / or, Method 5: Solution heating-slow cooling HSC method, including the following steps: Weigh the thiophene pyrimidine derivative methanesulfonate sample into a glass bottle, place the sample bottle on a magnetically heated stirrer, water bath temperature 45-55℃, rotation speed 200-250rpm, add solvent, heat to promote sample dissolution, keep warm for 10-20min, filter each solution with a 0.45-0.5μm filter membrane while hot, transfer the filtrate to a new sample bottle, and slowly cool to room temperature at a rate of 6℃ / h. Centrifuge the solvent system that precipitates a large amount of solid, remove the solid, evaporate the solvent at room temperature, and obtain the sample as crystal form I; if very little or no precipitation occurs, remove the sample, protect it from light at room temperature, magnetically stir until solid precipitates, then centrifuge and collect the precipitated solid, and obtain the sample as crystal form I.

14. The preparation method according to claim 13, characterized in that, In Method 1, the solvent is selected from one or both of dichloromethane and methanol; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (80-100 mg): (6-70 ml); And / or, in Method 2, the solvent is selected from one or more of N,N-dimethylacetamide, methyl tert-butyl ether, acetonitrile, ethanol, acetone, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, n-heptane, 1,4-dioxane, 2-butanone, isopropanol, and dichloromethane; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (80-100 mg): (4-5 ml); And / or, in Method 3, the solvent is selected from one or more of dichloromethane, pure water, dimethyl sulfoxide, methanol, and N,N-dimethylacetamide; and / or, the antisolvent is selected from one or more of tetrahydrofuran, isopropanol, 1,4-dioxane, acetone, tetrahydrofuran, dichloromethane, ethyl acetate, isopropanol, acetonitrile, 2-butanone, ethyl acetate, isopropanol, isopropyl ether, ethyl acetate, and tetrahydrofuran; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent and antisolvent is (80-100 mg): (1-20 ml): (5-280 ml); and / or, the stirring time is 10-20 min; And / or, in Method 4, the solvent is selected from one or more of dimethyl sulfoxide, methanol and acetonitrile; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (50-100 mg): (0.9-30 ml); and / or, the stirring time is 10-20 min; And / or, in method five, the solvent is selected from one or both of methanol and ethanol; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is (50-60 mg): (1-10 ml); and / or, the stirring time is 10-20 min.

15. A method for preparing the crystal form of the thiophenepyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The preparation method of the crystal form II includes the following steps: placing the thiophene pyrimidine derivative methanesulfonate sample in a glass bottle, placing the sample bottle on a magnetically heated stirrer, water bath temperature of about 45-55℃, rotation speed of 200-250 rpm, adding solvent, heating to promote sample dissolution, keeping warm for 10-20 min, filtering the solution while hot through a 0.45-0.5 μm filter membrane, transferring the filtrate to a new sample bottle, slowly adding antisolvent dropwise to the glass bottle under magnetic stirring, centrifuging the solvent system with precipitated solid and collecting the solid, evaporating the solvent at room temperature, and obtaining the sample as the crystal form II; the solvent system without precipitated solid is allowed to cool naturally to room temperature and magnetically stirred. If no solid still precipitates, it is stirred open until solid precipitates, and the sample obtained is the crystal form II.

16. The preparation method according to claim 15, characterized in that, The solvent is selected from dichloromethane; and / or the antisolvent is selected from ethyl acetate; and / or the ratio of the thiophene pyrimidine derivative methanesulfonate, the solvent, and the antisolvent is 80 mg: 70 ml: 490 ml; and / or the stirring time is 10-20 min.

17. A method for preparing the crystal form of the thiophene pyrimidine derivative methanesulfonate as described in claim 2, characterized in that, The preparation method of the crystal form III includes the following steps: placing the thiophene pyrimidine derivative methanesulfonate sample in a glass bottle, placing the sample bottle on a magnetically heated stirrer, water bath temperature of about 45-55℃, rotation speed of 200-250 rpm, adding solvent, heating to promote sample dissolution, keeping warm for 10-20 min, filtering the solution while hot through a 0.45-0.5 μm filter membrane, transferring the filtrate to a new sample bottle, immediately placing the sample bottle in a -20℃ refrigerator, centrifuging the solvent system that precipitates a large amount of solid, removing the solid, evaporating the solvent at room temperature to obtain the crystal form III; if very little or no precipitation occurs, the sample is removed, protected from light at room temperature, magnetically stirred until solid precipitates, and then the precipitated solid is collected by centrifugation to obtain the crystal form III.

18. The preparation method according to claim 17, characterized in that, The solvent is selected from n-butanol; and / or, the ratio of the thiophene pyrimidine derivative methanesulfonate to the solvent is 50 mg: 40 ml; and / or, the stirring time is 10-20 min.

19. A drug / drug composition, characterized in that, The drug / drug composition comprises the crystal form of the thiophene pyrimidine derivative mesylate as described in any one of claims 1-12; further, the drug / drug composition also comprises a pharmaceutically acceptable carrier.

20. The crystal form of the thiophene pyrimidine derivative mesylate according to any one of claims 1-12, or the preparation method according to any one of claims 13-18, or the use of the drug / drug composition according to claim 19 in the preparation of a medicament for treating diseases caused by abnormal cell proliferation due to overexpression of EGFR.

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