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

Abstract

The application discloses a crystal form of a thiophene pyrimidine derivative and a preparation method and application thereof. The crystal form of the thiophene pyrimidine derivative comprises crystal form A, crystal form B, crystal form C, crystal form D, crystal form E, crystal form F, crystal form G and crystal form H. The application further discloses a medicine / medicine composition. The application further discloses application of the crystal form of the thiophene pyrimidine derivative 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 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, 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, EMBO J, 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, Sci Transl Med, 2011, 3, 75ra26; S. Kobayashi, N Engl J Med, 2005, 352, 786-792; W. Pao, PLoS 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, Proc Natl Acad Sci USA, 2008, 105, 2070-2075; Cancer Cell, 2007, 11, 217-227; M. Azam, Nat Struct Mol Biol, 2008, 15, 1109-1118; TA. Carter, Proc Natl Acad Sci USA, 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 derivatives are effective and selective inhibitors of mutant epidermal growth factor receptor (EGFR) tyrosine kinases without affecting wild-type EGFR, thus reducing side effects.

[0009] Pyrimidine derivatives, represented by osimertinib, are third-generation EGFR-TKIs that are effective and selective inhibitors of mutant epidermal growth factor receptor (EGFR) tyrosine kinases without affecting wild-type EGFR, thus reducing side effects.

[0010] 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.

[0011] 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

[0012] To address the shortcomings of existing technologies, the present invention aims to provide a crystal form of a thiophene pyrimidine derivative, 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, resulting in a product with a unique and stable crystal form, which has been successfully used in preclinical and clinical studies.

[0013] This invention provides a crystal form of a thiophene pyrimidine derivative, the structural formula of which is shown in formula (I):

[0014]

[0015] The crystal forms of the thiophene pyrimidine derivatives are selected from the following group: crystal form A, crystal form B, crystal form C, crystal form D, crystal form E, crystal form F, crystal form G, and crystal form H, wherein crystal form H is the dominant crystal form.

[0016] In some embodiments, the X-ray powder diffraction pattern of crystal form H of the thiophene pyrimidine derivative has characteristic peaks at 2θ angles of 8.626, 10.658, 12.532, 19.409, 21.402, and 27.008.

[0017] In some embodiments, the crystal form is crystal form A.

[0018] In some embodiments, the crystal form A of the thiophene pyrimidine derivative has essentially the following properties: Figure 24 The X-ray powder diffraction pattern shown.

[0019] In some embodiments, the crystal form A of the thiophene pyrimidine derivative has essentially the following properties: Figure 25 The DSC and TGA spectra shown are shown.

[0020] In some embodiments, the crystal form A of the thiophene pyrimidine derivative has essentially the following properties: Figure 26 The XRPD images shown are before and after heating.

[0021] In some embodiments, the crystal form A of the thiophene pyrimidine derivative has essentially the following properties: Figure 27 shown 1 HNMR image.

[0022] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form A of the thiophene pyrimidine derivative includes an endothermic peak at 172.5 °C.

[0023] In some embodiments, the crystal form is crystal form B.

[0024] In some embodiments, the crystal form B of the thiophene pyrimidine derivative has essentially the following properties: Figure 28 The X-ray powder diffraction pattern shown.

[0025] In some embodiments, the crystal form B of the thiophene pyrimidine derivative has essentially the following properties: Figure 29 The DSC and TGA spectra shown are shown.

[0026] In some embodiments, the crystal form B of the thiophene pyrimidine derivative has essentially the following properties: Figure 30 shown 1 HNMR image.

[0027] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form B of the thiophene pyrimidine derivative includes an endothermic peak at 170.9 °C.

[0028] In some embodiments, the crystal form is crystal form C.

[0029] In some embodiments, the crystal form C of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 31 The X-ray powder diffraction pattern shown.

[0030] In some embodiments, the crystal form C of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 32 The DSC and TGA spectra shown are shown.

[0031] In some embodiments, the crystal form C of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 33 The XRPD images shown are before and after heating.

[0032] In some embodiments, the crystal form C of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 34 shown 1 HNMR image.

[0033] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form C of the thiophene pyrimidine derivative includes an endothermic peak at 172.6 °C.

[0034] In some embodiments, the crystal form is crystal form D.

[0035] In some embodiments, the crystal form D of the thiophene pyrimidine derivative has essentially the following properties: Figure 35 The X-ray powder diffraction pattern shown.

[0036] In some embodiments, the crystal form D of the thiophene pyrimidine derivative has essentially the following properties: Figure 36 The DSC and TGA spectra shown are shown.

[0037] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form D of the thiophene pyrimidine derivative includes an endothermic peak at 175.0 °C.

[0038] In some embodiments, the crystal form is crystal form E.

[0039] In some embodiments, the crystal form E of the thiophene pyrimidine derivative has essentially the following properties: Figure 37 The X-ray powder diffraction pattern shown.

[0040] In some embodiments, the crystal form E of the thiophene pyrimidine derivative has essentially the following properties: Figure 38 The DSC and TGA spectra shown are shown.

[0041] In some embodiments, the crystal form E of the thiophene pyrimidine derivative has essentially the following properties: Figure 39 shown 1 HNMR image.

[0042] In some embodiments, the crystal form E of the thiophene pyrimidine derivative has essentially the following properties: Figure 40 The XRPD images shown are before and after heating.

[0043] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form E of the thiophene pyrimidine derivative includes an endothermic peak at 79.6 °C.

[0044] In some embodiments, the crystal form is crystal form F.

[0045] In some embodiments, the crystal form F of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 41 The X-ray powder diffraction pattern shown.

[0046] In some embodiments, the crystal form F of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 42 The DSC and TGA spectra shown are shown.

[0047] In some embodiments, the crystal form F of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 43 The XRPD images shown are before and after heating.

[0048] In some embodiments, the crystal form F of the thiophene pyrimidine derivative has essentially the following characteristics: Figure 44 shown 1 HNMR image.

[0049] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form F of the thiophene pyrimidine derivative includes an endothermic peak at 173.4 °C.

[0050] In some embodiments, the crystal form is crystal form G.

[0051] In some embodiments, the crystal form G of the thiophene pyrimidine derivative has essentially the following properties: Figure 45 The X-ray powder diffraction pattern shown.

[0052] In some embodiments, the crystal form G of the thiophene pyrimidine derivative has essentially the following properties: Figure 46 The DSC and TGA spectra shown are shown.

[0053] In some embodiments, the crystal form G of the thiophene pyrimidine derivative has essentially the following properties: Figure 47 shown 1 HNMR image.

[0054] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form G of the thiophene pyrimidine derivative includes an endothermic peak at 149.8 °C.

[0055] In some embodiments, the crystal form is crystal form H.

[0056] In some embodiments, the crystal form H of the thiophene pyrimidine derivative has essentially the following properties: Figure 49 The X-ray powder diffraction pattern shown.

[0057] In some embodiments, the crystal form H of the thiophene pyrimidine derivative has essentially the following properties: Figure 50 The XRPD peak finding report shown.

[0058] In some embodiments, the crystal form H of the thiophene pyrimidine derivative has essentially the following properties: Figure 51 The DSC and TGA spectra shown are shown.

[0059] In some embodiments, the differential scanning calorimetry (DSC) curve of crystal form H of the thiophene pyrimidine derivative includes an endothermic peak at 171.0 °C.

[0060] Specifically, the thiophene pyrimidine derivative has a crystal form H that is an anhydrous crystal with a single melting point and characteristic XRPD diffraction peaks; it exhibits relatively good physicochemical stability at 25℃ / 60%RH and 40℃ / 75%RH; it has good thermal stability at 80℃; and it has low hygroscopicity at a constant temperature of 25℃ when the humidity increases from 0%RH to 80%RH.

[0061] The present invention also provides a method for preparing crystal form H of the thiophene pyrimidine derivative as described above. The preparation method includes the following steps: using the received amorphous thiophene pyrimidine derivative as the starting material, preparing a suspension in a solvent, stirring at 20-55°C for 1 to 5 days, and then separating the solid to obtain crystal form H of the thiophene pyrimidine derivative.

[0062] Specifically, the solvent includes one or more of isopropanol, dichloromethane, ethyl acetate, etc.; preferably, it is isopropanol and ethyl acetate.

[0063] Specifically, the stirring temperature is 20-55℃; preferably, it is 25℃.

[0064] Specifically, the stirring time is 1 to 5 days; preferably, it is 2 days.

[0065] Specifically, the ratio of the thiophene pyrimidine derivative to the solvent is (15-100) mg:(0.5-2) mL; preferably, it is 100 mg:2 mL.

[0066] The preparation method of the crystal form A includes the following steps: crystal form A is obtained by adding n-heptane dropwise to an ethanol (EtOH) solution of a thiophene pyrimidine derivative of the starting sample and crystallizing.

[0067] The preparation method of the crystal form B includes one or more of the following methods: antisolvent addition, slow evaporation, slow cooling, gas-solid permeation, gas-liquid diffusion, and suspension stirring. Preferably, the preparation method includes the following steps: preparing the crystal form B by suspending and stirring the starting sample thiophene pyrimidine derivative in isopropanol (IPA) at room temperature.

[0068] The preparation method of the crystal form C includes one or more of slow volatilization, gas-solid infiltration, and polymer-induced methods. Preferably, the preparation method includes the following steps: obtaining the crystal form C by slowly volatilizing a methanol (MeOH) solution of a thiophene pyrimidine derivative of the starting sample at room temperature.

[0069] The method for preparing the crystal form D is a room temperature suspension and stirring method, which includes the following steps: obtaining the crystal form D by suspending and stirring the starting sample thiophene pyrimidine derivative in IPA / H2O (aw~0.8) at room temperature;

[0070] The method for preparing the crystal form E is a room temperature suspension and stirring method, which includes the following steps: obtaining the crystal form E by suspending and stirring the starting sample thiophene pyrimidine derivative in MeOH / isopropyl ester (IPAc) (1:2, v / v) at room temperature;

[0071] The method for preparing the crystal form F is a room temperature suspension and stirring method, which includes the following steps: obtaining the crystal form F by suspending and stirring the starting sample thiophene pyrimidine derivative in EtOH / H2O (1:1, v / v) at room temperature;

[0072] The preparation method of the crystal form G is a suspension stirring method at room temperature or 50°C. Preferably, the preparation method includes the following steps: obtaining the crystal form G by suspending and stirring the starting sample thiophene pyrimidine derivative in acetonitrile (ACN) / ethyl acetate (EtOAc) (1:1, v / v) at room temperature.

[0073] Specifically, the preparation method, preparation steps, and reaction conditions of the thiophene pyrimidine derivative crystal form AG of the present invention are illustrated in Example 1.

[0074] The crystal form AH of the thiophene pyrimidine derivative described in this invention is proposed for the first time.

[0075] The present invention provides a drug / drug composition comprising the crystal form of a thiophene pyrimidine derivative as described above.

[0076] Furthermore, the drug / drug composition comprises a pharmaceutically acceptable carrier.

[0077] 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.

[0078] 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.

[0079] Specifically, the drug / drug composition is administered orally, by injection, nasal administration, transdermal administration, or mucosal administration.

[0080] 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.

[0081] Specifically, the drug / drug composition includes liquid dosage forms, gaseous dosage forms, solid dosage forms, and semi-solid dosage forms.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] 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)).

[0091] 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.

[0092] 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.

[0093] 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.

[0094] 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.

[0095] The pharmaceutical compositions described in this invention may be included in a container, package, or dispenser along with instructions for administration.

[0096] 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.

[0097] The present invention also provides the use of the crystal form of the thiophene pyrimidine derivative as described above, or the preparation method as described above, or 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.

[0098] The beneficial effects of this invention include: through the research and development of patented thiophene pyrimidine derivatives, this invention has successfully screened out the crystal forms of stable thiophene pyrimidine derivatives for the first time, and these derivatives have been successfully used in preclinical and clinical studies. Attached Figure Description

[0099] 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.

[0100] Figure 1 This is a diagram showing the interconversion relationships between polymorphs of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0101] Figure 2 This is a summary of the characterization results of the polymorphisms of the thiophene pyrimidine derivatives in Example 1 of the present invention;

[0102] Figure 3 This is the XRPD image of the starting sample thiophene pyrimidine derivative (806131-05-A) in Example 1 of this invention;

[0103] Figure 4 This is a TGA / DSC chromatogram of the starting sample thiophene pyrimidine derivative (806131-05-A) in Example 1 of this invention;

[0104] Figure 5 This is the XRPD image of the starting sample thiophene pyrimidine derivative (806131-05-A) in Example 1 of this invention after heating to 60°C;

[0105] Figure 6This is a summary graph of the pKa results calculated using the Yasuda-Shedlovsky extrapolation method for the thiophene pyrimidine derivatives of the starting sample in Example 1 of this invention;

[0106] Figure 7 The pKa diagram of the thiophene pyrimidine derivative of the starting sample in Example 1 of this invention was calculated using the Yasuda-Shedlovsky extrapolation method.

[0107] Figure 8 This is the approximate solubility of the thiophene pyrimidine derivative (806131-05-A) in the starting sample of Example 1 of this invention at room temperature;

[0108] Figure 9 This is a summary of the results of the repeated preparation of crystal form D of thiophene pyrimidine derivatives in Example 1 of this invention;

[0109] Figure 10 This is an XRPD overlay of the solid D crystal form of the thiophene pyrimidine derivative attempted to be prepared repeatedly in Example 1 of this invention;

[0110] Figure 11 This is a suspension competition stirring test between thiophene pyrimidine derivatives and their crystal-free B / H components in Example 1 of this invention;

[0111] Figure 12 This is an XRPD overlay (I / II) of the crystalline B / H suspension competitive separation solids of the thiophene pyrimidine derivative in Example 1 of this invention;

[0112] Figure 13 This is an XRPD overlay (II / II) of the crystalline B / H suspension competitive separation solids of the thiophene pyrimidine derivative in Example 1 of this invention;

[0113] Figure 14 This is a summary of the polymorph screening test of thiophene pyrimidine derivatives in Example 1 of the present invention;

[0114] Figure 15 This is a summary of the antisolvent addition experiment in Example 1 of the present invention;

[0115] Figure 16 This is a summary of the slow volatilization test in Example 1 of the present invention;

[0116] Figure 17 This is a summary of the slow cooling experiment in Embodiment 1 of the present invention;

[0117] Figure 18 This is a summary of the room temperature suspension stirring test in Example 1 of the present invention;

[0118] Figure 19 This is a summary of the 50°C suspension stirring test in Example 1 of the present invention;

[0119] Figure 20 This is a summary of the gas-solid diffusion experiment in Example 1 of the present invention;

[0120] Figure 21 This is a summary of the gas-liquid diffusion experiment in Example 1 of the present invention;

[0121] Figure 22 This is a summary of the polymer-induced experiment in Example 1 of the present invention;

[0122] Figure 23 These are XRPD comparison images of the polymorphs (polymorphs A to H) of the thiophene pyrimidine derivatives in Example 1 of this invention;

[0123] Figure 24 This is the XRPD image of crystal form A (806131-16-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0124] Figure 25 This is the TGA / DSC image of crystal form A (806131-16-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0125] Figure 26 These are XRPD images of crystal form A (806131-16-Al) of the thiophene pyrimidine derivative in Example 1 of this invention before and after heating;

[0126] Figure 27 It is the crystal form A (806131-16-Al) of the thiophene pyrimidine derivative in Example 1 of this invention. 1 HNMR spectrum;

[0127] Figure 28 This is the XRPD image of crystal form B (806131-17-A1) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0128] Figure 29 This is the TGA / DSC image of crystal form B (806131-17-A1) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0129] Figure 30 It is the crystal form B (806131-17-A1) of the thiophene pyrimidine derivative in Example 1 of this invention. 1 HNMR spectrum;

[0130] Figure 31 This is the XRPD image of crystal form C (806131-18-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0131] Figure 32 This is the TGA / DSC image of crystal form C (806131-18-A1) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0132] Figure 33 These are XRPD images of the thiophene pyrimidine derivative C (806131-18-Al) before and after heating in Example 1 of this invention;

[0133] Figure 34 It is the crystal form C (806131-18-A1) of the thiophene pyrimidine derivative in Example 1 of this invention. 1 HNMR spectrum;

[0134] Figure 35 This is the XRPD image of crystal form D (806131-10-A18) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0135] Figure 36 This is the TGA / DSC image of crystal form D (806131-10-A18) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0136] Figure 37 This is the XRPD image of crystal form E (806131-17-A3) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0137] Figure 38 This is the TGA / DSC image of crystal form E (806131-17-A3) of the thiophene pyrimidine derivative in Example 1 of this invention;

[0138] Figure 39 It is the crystal form E (806131-17-A3) of the thiophene pyrimidine derivative in Example 1 of this invention. 1 HNMR spectrum;

[0139] Figure 40 These are XRPD images of the thiophene pyrimidine derivative E (806131-17-A3) before and after heating in Example 1 of this invention;

[0140] Figure 41 This is the XRPD image of the crystal form F (806131-17-A4) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0141] Figure 42 This is the TGA / DSC image of crystal form F (806131-17-A4) of the thiophene pyrimidine derivative in Example 1 of this invention;

[0142] Figure 43 These are XRPD images of the thiophene pyrimidine derivative F(806131-17-A4) before and after heating in Example 1 of this invention;

[0143] Figure 44 It is the crystal form F(806131-17-A4) of the thiophene pyrimidine derivative in Example 1 of this invention. 1HNMR spectrum;

[0144] Figure 45 This is the XRPD image of the crystal form G (806131-17-A5) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0145] Figure 46 This is the TGA / DSC image of crystal form G (806131-17-A5) of the thiophene pyrimidine derivative in Example 1 of this invention;

[0146] Figure 47 It is the crystal form G (806131-17-A5) of the thiophene pyrimidine derivative in Example 1 of this invention. 1 HNMR spectrum;

[0147] Figure 48 This is the XRPD image of the crystal form H (806131-21-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0148] Figure 49 This is the XRPD image (with numbers) of the crystal form H (806131-21-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0149] Figure 50 This is the XRPD peak finding report of crystal form H of the thiophene pyrimidine derivative in Example 1 of this invention;

[0150] Figure 51 This is the TGA / DSC image of the crystal form H (806131-21-A1) of the thiophene pyrimidine derivative in Example 1 of this invention;

[0151] Figure 52 This is a competitive stirring test of the suspension of thiophene pyrimidine derivatives A / B / C / G in Example 1 of this invention;

[0152] Figure 53 This is an XRPD overlay of the crystalline A / B / C / G suspension competitively separated solids of the thiophene pyrimidine derivative in Example 1 of the present invention, wherein, as indicated by the black spikes, the solids separated from IPA / room temperature conditions contain a small amount of peaks of type G;

[0153] Figure 54 This is a summary of the physicochemical stability results of the thiophene pyrimidine derivative in Example 1 of this invention, crystal form H (806131-21-Al).

[0154] Figure 55 This is a comparison table of impurities regarding the physicochemical stability of the thiophene pyrimidine derivative in crystal form H (806131-21-Al) in Example 1 of this invention;

[0155] Figure 56Comparison of HPLC chromatograms of the physicochemical stability of crystal form H(806131-21-A1) of the thiophenepyrimidine derivative in Example 1 of the present invention;

[0156] Figure 57 This is an XRPD overlay image of the crystal form H (806131-21-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention before and after stable placement;

[0157] Figure 58 This is the DVS diagram of the crystal form H (806131-21-Al) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0158] Figure 59 This is a comparison diagram of the crystal form H (806131-21-A1) of the thiophene pyrimidine derivative in Example 1 of the present invention before and after DVS testing;

[0159] Figure 60 This is a PLM diagram of the crystal form H (806131-21-A1) of the thiophene pyrimidine derivative in Example 1 of the present invention;

[0160] Figure 61 This is an evaluation of the equilibrium solubility of the thiophene pyrimidine derivatives A / B / C / G / H in pure water over 24 hours in Example 1 of this invention.

[0161] Figure 62 This is a comparison of XRPD values ​​of crystal forms A / B / C of the thiophene pyrimidine derivatives in Example 1 of the present invention after equilibration in pure water for 24 hours;

[0162] Figure 63 This is a comparison of XRPD values ​​of the thiophene pyrimidine derivative G / H in Example 1 of this invention after equilibration in pure water for 24 hours;

[0163] Figure 64 This is a summary of the results of repeated preparation of the dominant crystal form H of the thiophene pyrimidine derivative in Example 1 of this invention;

[0164] Figure 65 This is an XRPD image of the dominant crystalline form H solid obtained by repeated preparation of the thiophene pyrimidine derivative in Example 1 of this invention;

[0165] Figure 66 This is a summary of the results of repeated preparation of the dominant crystal form H (100 mg) in Example 1 of the present invention;

[0166] Figure 67 This is the XRPD diagram of the repeated preparation of the dominant crystalline form H solid (100 mg) in Example 1 of this invention;

[0167] Figure 68This is the XRD pattern of the crystalline compound JFAN-1001 (crystalline form H) of the thiophene pyrimidine derivative in Example 1 of this invention.

[0168] In the accompanying drawings of this invention, intensity represents strength; counts represent counts; Heat represents heat. Flow indicates heat flux; Temperature indicates temperature; Solvent indicates solvent; Extrapolation indicates extrapolation method; Dielectric constant indicates dielectric constant; MeOH indicates methanol; EtOH indicates ethanol; IPA indicates isopropanol; Acetone indicates acetone; MIBK indicates methyl isobutyl ketone; EtOAc indicates ethyl acetate; IPAc indicates isopropyl acetate; MTBE indicates methyl tert-butyl ether; THF indicates tetrahydrofuran; 2-MeTHF indicates 2-methyltetrahydrofuran; n-Heptane indicates n-heptane; water indicates water; Type indicates crystal form; ACN indicates acetonitrile; DMSO indicates dimethyl sulfoxide; 1,4-dioxane indicates 1,4-dioxane; DCM indicates dichloromethane; Anisole indicates anisole; NMP indicates N-methylpyrrolidone; CHCl3 indicates chloroform / trichloromethane; toluene indicates toluene; n-heptane indicates n-heptane; Weight indicates weight; Change in Mass indicates mass change; Target RH represents the target humidity; Isotherm Plot represents an isotherm plot. Detailed Implementation

[0169] 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.

[0170] 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.

[0171] 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.

[0172] This invention discloses a crystalline form of a thiophene pyrimidine derivative, its preparation method, and its application. The crystalline forms of the thiophene pyrimidine derivative include crystalline forms A, B, C, D, E, F, G, and H. This invention also discloses a drug / drug composition. Furthermore, this invention discloses the application of the crystalline forms of the thiophene pyrimidine derivative 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.

[0173] Unless otherwise specified, the experimental materials used in the examples are all conventional biochemical reagents.

[0174] Example 1: Crystal form screening experiment of thiophene pyrimidine derivatives

[0175] This invention uses thiophene pyrimidine derivatives as starting samples and employs various screening methods (including: antisolvent addition, slow evaporation, slow cooling, room temperature / 50°C suspension stirring, gas-solid permeation, gas-liquid permeation, and polymer induction) to conduct a total of 101 polymorph screening experiments. Based on X-ray powder diffraction (XRPD) characterization of the separated solids and subsequent identification and evaluation, a total of 8 different polymorphs were discovered, including 5 amorphous forms A / B / C / G / H, 2 solvate forms E / F, and 1 hydrate form D (which could not be replicated). All discovered polymorphs were characterized by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), and the results are summarized as follows: Figure 2 As shown in the figure. Heating experiments under nitrogen protection showed that crystalline forms A / C / F all transformed into crystalline form B upon heating; crystalline form E transformed into an amorphous form upon heating. Suspension and stirring results in different temperatures and solvent systems indicated that amorphous form H was more stable than amorphous forms A / B / C / G at room temperature (25±3℃) and 50℃. The transformation relationships between polymorphs are shown in the figure. Figure 1 As shown.

[0176] In subsequent property evaluations, the physicochemical stability, hygroscopicity, and particle size of the dominant crystalline form H were tested under different temperatures and humidity levels. Physicochemical stability tests showed that crystalline form H exhibited relatively good physicochemical stability at 25℃ / 60%RH and 40℃ / 75%RH, with a purity / initial purity (area %) of 99.9–100.1% after one week of storage. After 36 hours of storage at 80℃, the purity was 99.0% of the initial purity, indicating slight degradation. Dynamic moisture adsorption (DVS) tests showed that at a constant temperature of 25℃, the crystalline form H sample gained 1.0% weight due to moisture absorption when the humidity increased from 0%RH to 80%RH, indicating that crystalline form H has slight hygroscopicity. Polarizing microscopy (PLM) testing revealed that the particles of this batch of crystalline form H were small (0–10 μm) and exhibited agglomeration.

[0177] Based on the above screening and evaluation, crystal form H is recommended as the crystal form of the superior thiophene pyrimidine derivative, that is, the crystal form of the thiophene pyrimidine derivative described in this invention, for further development.

[0178] The crystal forms A to H of the thiophene pyrimidine derivatives prepared in the crystal form screening experiment of the present invention were characterized by the following detection methods:

[0179] i) 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] ii) Powder X-ray diffraction analysis (XRPD)

[0182] The light source was Cu K, the X-ray intensity was 40 kV / 40 mA, 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] iii) 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] iv) 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] v) One-dimensional proton nuclear magnetic resonance spectrum ( 1 H NMR)

[0188] Weigh approximately 5 mg of sample into an NMR tube, add 0.6 mL of deuterated dimethyl sulfoxide (DMSO d6) 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] 1. Starting sample

[0190] 1.1 Physical Characterization

[0191] In this embodiment of the invention, a thiophene pyrimidine derivative was used as the starting sample. The sample was characterized in solid state using XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry), and TGA (thermogravimetric analysis). The XRPD results of the sample are as follows: Figure 3 As shown, this batch of samples is amorphous. Figure 4 As shown, TGA results indicate that the sample lost 4.3% of its weight before reaching 150℃. DSC data show two overlapping endothermic peaks at 68.7℃ and 97.7℃ (peak temperatures). After heating the sample to 60℃ and then cooling it to room temperature under nitrogen protection, the resulting solid remained amorphous, and its XRPD is as follows: Figure 5 As shown. When heated to 150°C and then cooled to room temperature, a molten solid is obtained.

[0192] The pKa of the thiophene pyrimidine derivatives in the samples was tested using a physicochemical constant analyzer (Pion Inc., Sirius T3). Figure 6 , 7 The results show that two pKa values ​​were measured in this invention (pKa1 = 4.15, pKa2 = 7.41). During the test, when the pH was greater than 8, solid precipitation was observed, which affected the pKa measurement; therefore, the pKa2 result is for reference only.

[0193] 1.2 Approximate solubility (to provide available solvents for screening)

[0194] This invention used 20 solvents to roughly determine the solubility of the starting sample, a thiophene pyrimidine derivative, at room temperature. In the experiment, approximately 2 mg of solid was weighed into a 3 mL vial, and 100 μL of the corresponding solvent was added dropwise each time until dissolved. Addition was stopped when the total solvent volume reached 2 mL and the solid had not completely dissolved. Solubility data are shown below. Figure 8 As shown.

[0195] 2. Attempt to repeatedly prepare crystal form D of the thiophene pyrimidine derivative.

[0196] In the batch screening tests of this invention, crystal form D was obtained by stirring the amorphous thiophene pyrimidine derivatives of the starting sample at room temperature in isopropanol (IPA) / H2O (aw~0.8), acetone (acetone) / H2O (1:1), and pure water systems. The three experimental conditions were repeated, and attempts were made to replicate the preparation of crystal form D by changing the temperature range, as follows: Figure 9 , 10 As shown, the solid obtained were crystalline forms B, H, or amorphous thiophene pyrimidine derivatives, but crystalline form D of the thiophene pyrimidine derivative was not successfully obtained.

[0197] 3. Comparison of thermodynamic stability of B / H crystal forms

[0198] This invention utilizes heating under nitrogen protection to transform the crystal forms A / C / F of thiophenepyrimidine derivatives into crystal form B. To further confirm the thermodynamic stability relationship between crystal form B and crystal form H, samples of both crystal forms (crystal form B + crystal form H) were suspended and stirred in different solvent systems at room temperature (25℃±3℃) and 50℃. The experimental results are as follows. Figure 11 And XRPD results (such as Figure 12 , 13 As shown in the figure, the amorphous form B transformed into crystalline form H after stirring in n-heptane and IPA / water (aw ~ 0.4 / 0.8), indicating that crystalline form H is more stable than crystalline form B between room temperature and 50°C.

[0199] 4. Polymorph screening test

[0200] This invention employed various crystallization methods and set up a total of 101 polymorph screening tests. The specific test methods and results are summarized as follows: Figure 14 As shown.

[0201] 4.1 Antisolvent addition test

[0202] This invention includes 16 antisolvent addition tests. Approximately 20 mg of each starting sample of the thiophene pyrimidine derivative is weighed into a 20 mL vial and dissolved in 0.5-2.0 mL of a good solvent (if incompletely dissolved, it is filtered through a 45 μm PTFE filter). The solvent is then added dropwise to the clear solution as follows: Figure 15 Add the antisolvent dropwise while stirring until solid precipitates. If no solid precipitates after adding approximately 10.0 mL of antisolvent, stop the experiment, collect the solid, and perform XRPD testing. The experimental results are as follows: Figure 15 As shown, the antisolvent addition experiment yielded crystal forms A / B and amorphous forms.

[0203] 4.2 Slow evaporation

[0204] This invention includes 10 slow evaporation tests. Approximately 15 mg of each thiophene pyrimidine derivative sample was weighed into a 3 mL vial, and 1.0 mL of solvent was added. The sample was sonicated or shaken to dissolve it completely as much as possible. After filtration, the supernatant was collected. The vial was sealed with sealing film and 3-5 pinholes were pricked in the film. It was then allowed to slowly evaporate at room temperature. The resulting solid was collected and subjected to XRPD testing. The test results are as follows: Figure 16 As shown, crystal forms B / C and amorphous forms were obtained in the slow volatilization crystallization experiment.

[0205] 4.3 Slow cooling

[0206] This invention includes 10 slow cooling experiments. Approximately 15 mg of each initial sample of the thiophene pyrimidine derivative was weighed into a 1.5 mL vial, and 0.75 mL of [amount missing] was added. Figure 17 The solvent shown was stirred at 50°C for approximately 2 hours, then filtered to obtain the supernatant. The supernatant was cooled from 50°C to 5°C at a rate of 0.1°C / min and held at 5°C. The precipitated solid was collected and subjected to XRPD testing. Samples without precipitated solids were transferred to room temperature for evaporation. The experimental results are shown below. Figure 17 The slow cooling experiment yielded crystalline form B and amorphous form.

[0207] 4.4 Room temperature suspension stirring

[0208] This invention included 18 room temperature suspension stirring tests. Approximately 15 mg of each initial sample of the thiophene pyrimidine derivative was weighed into a 1.5 mL glass vial, and 0.5 mL of [amount missing] was added to each vial. Figure 18 The solvents listed are used to prepare suspensions. These suspensions are then stirred at room temperature for approximately 6 days. The solids are collected by centrifugation and subjected to XRPD testing. The experimental results are shown below. Figure 18 As shown, the suspension stirring test yielded crystal forms B / D / E / F / G and amorphous forms.

[0209] 4.550℃ Suspension stirring

[0210] This invention included 14 suspension stirring tests at 50°C. Approximately 15 mg of each initial sample of the thiophene pyrimidine derivative was weighed into a 1.5 mL glass vial, and 0.5 mL of [amount missing] was added to each vial. Figure 19 The solvents listed in the figure were used to obtain a suspension, which was stirred at 50°C for about 6 days. The solid was then collected by centrifugation and subjected to XRPD testing. The experimental results are as follows: Figure 19 As shown, crystalline B / G and amorphous forms were obtained in the suspension stirring test.

[0211] 4.6 Gas-solid permeation

[0212] This invention comprises 11 gas-solid diffusion experiments. Approximately 10 mg of each thiophene pyrimidine derivative of the starting sample was weighed into a 3 mL vial. A separate 20 mL vial was taken, and approximately 4 mL of solvent was added to it. The 3 mL vial was placed open inside the 20 mL vial, which was then sealed and allowed to stand at room temperature for 9 days. The solid was collected and XRPD analysis was performed. The experimental results are as follows: Figure 20 As shown, crystalline B / C and amorphous forms were obtained in the gas-solid permeation test.

[0213] 4.7 Gas-liquid diffusion

[0214] This invention comprises 12 gas-liquid diffusion experiments. Approximately 15 mg of each thiophene pyrimidine derivative of the initial sample is weighed and dissolved in 0.5 mL of a good solvent. The supernatant is filtered and transferred to a 3 mL vial. A separate 20 mL vial is prepared, and approximately 4 mL of the antisolvent is added. The 3 mL vial is placed open inside the 20 mL vial, which is then sealed and allowed to stand at room temperature. When solid precipitation is observed, the solid is removed and XRPD is measured. If no solid precipitation occurs, the solid is removed and allowed to evaporate at room temperature until the solvent is completely evaporated. The solid is then collected and XRPD is measured. The experimental results are as follows: Figure 21 As shown, gas-liquid diffusion experiments yielded crystalline form B and amorphous forms.

[0215] 4.8 Polymer-induced

[0216] This invention includes 10 polymer-induced assays. Approximately 10 mg of each starting sample of the thiophene pyrimidine derivative was dissolved in 1.0 mL of a good solvent and filtered to obtain a clear solution. ~2 mg of the corresponding polymer mixture was added to each solution, and the vial was sealed with sealing film, with 3-5 pinholes punched in the film. The vial was then allowed to slowly evaporate at room temperature. The resulting solid was collected and subjected to XRPD testing. The experimental results are as follows: Figure 22 As shown, crystalline form C and amorphous form were obtained in the polymer-induced crystallization experiment.

[0217] 5. Screening and characterization of the crystal forms of thiophene pyrimidine derivatives prepared in the screening experiments of this invention.

[0218] Based on the XRPD characterization and subsequent identification and evaluation of the crystal forms of solid thiophene pyrimidine derivatives obtained in the screening experiments of this invention, a total of 8 polymorphs of thiophene pyrimidine derivatives were discovered, named crystal forms A to H respectively. The XRPD comparison results are as follows: Figure 23 As shown.

[0219] 5.1 Crystal form A

[0220] The crystal form A sample of this invention was obtained by crystallization of n-heptane by adding n-heptane dropwise to an ethanol (EtOH) solution of a thiophene pyrimidine derivative of the starting sample. Its XRPD results are as follows: Figure 24 As shown. The TGA / DSC results for this batch of samples ( Figure 25 The XRPD results show a weight loss of 1.6% when heated to 150℃; there is an endothermic peak at 94.4℃ (peak temperature), an exothermic peak at 119.7℃ (peak temperature), and a sharp endothermic peak at 168.2℃ (initial temperature). Under nitrogen protection, heating the sample to 95℃ and then cooling it to room temperature caused crystal form A to transform into crystal form B, as shown in the XRPD results. Figure 26 As shown. Meanwhile, the 1H NMR results for crystal form A ( Figure 27 EtOH was not detected in the sample. Based on the small weight loss of TGA before decomposition upon heating, crystal form A is presumed to be the amorphous form.

[0221] 5.2 Crystal form B

[0222] The crystal form B of this invention can be obtained through various methods, including antisolvent addition, slow evaporation, slow cooling, gas-solid permeation, gas-liquid diffusion, and suspension stirring. The crystal form B sample was prepared by suspending and stirring a starting sample of thiophene pyrimidine derivative in isopropanol (IPA) at room temperature. XRPD results are shown below. Figure 28 As shown. The TGA / DSC results for this batch of samples ( Figure 29 The results show a weight loss of 1.3% when heated to 150°C; there is a sharp endothermic peak at 164.3°C (initial temperature). 1 H NMR results ( Figure 30 The results showed that the crystal form B sample contained a small amount of residual IPA (API-active drug ingredient / IPA molar ratio 1:0.06, corresponding to a mass ratio of 0.63 wt%). Based on the small TGA weight loss before decomposition upon heating and the absence of thermal changes observed before melting, crystal form B is presumed to be an amorphous form.

[0223] 5.3 Crystal form C

[0224] The crystalline form C of this invention can be obtained through various methods, including slow evaporation, gas-solid infiltration, and polymer induction. The crystalline form C sample was obtained by slowly evaporating a methanol (MeOH) solution of a thiophene pyrimidine derivative from the starting sample at room temperature. Its XRPD results are as follows: Figure 31 As shown. The TGA / DSC results for this batch of samples ( Figure 32 The reading shows a weight loss of 1.3% when heated to 150°C; Figure 33 As shown, a sharp endothermic peak appears at 167.7℃ (initial temperature), preceded by multiple endothermic / exothermic peaks. Under nitrogen protection, heating the crystalline C sample to 110℃ and cooling to room temperature resulted in a characteristic peak indicating the transformation of crystalline C into an amorphous form mixed with a small amount of crystalline C. When heated to 155℃ and cooled to room temperature, the resulting amorphous form exhibited characteristic peaks mixed with a small amount of crystalline B, suggesting that crystalline C can be transformed into crystalline B upon heating. Simultaneously, the crystalline C sample... 1 H NMR results ( Figure 34 MeOH was not detected in the sample. Based on the small weight loss of TGA before decomposition upon heating, crystal form C is presumed to be an amorphous form.

[0225] 5.4 Crystal form D

[0226] Crystal form D of this invention can be obtained by room temperature suspension stirring. Crystal form D is obtained by suspending and stirring the starting sample thiophene pyrimidine derivative in IPA / H2O (aw~0.8) at room temperature. The XRPD results are as follows. Figure 35 As shown. The TGA / DSC results for this batch of crystal form D samples ( Figure 36The results showed a weight loss of 5.8% upon heating to 150°C; a sharp endothermic peak was observed at 172.9°C (initial temperature), following several previous endothermic / exothermic peaks. Attempts to repeatedly prepare crystal form D at room temperature and 50°C using IPA / H₂O (water activity (aw) ~0.8), acetone / H₂O (1:1, v / v), and pure water with suspension and stirring were unsuccessful (the resulting solids were crystal form B, crystal form H, or amorphous). Please refer to Section 2 of the Examples of this Invention for experimental conditions and results. Since crystal form D was obtained in solvent systems with high water activity, it is presumed to be a hydrate (based on the thermal signal in the DSC, it is inferred that crystal form D will transform into crystal form B during heating).

[0227] 5.5 Crystal Form E

[0228] Crystal form E of this invention can be obtained by room temperature suspension and stirring. Crystal form E is obtained by suspending and stirring the starting sample thiophene pyrimidine derivative in MeOH / isopropyl ester (IPAc) (1:2, v / v) at room temperature, and its XRPD results are as follows. Figure 37 As shown. The TGA / DSC of this batch of samples ( Figure 38 The sample showed an 8.5% weight loss upon heating to 100℃; an endothermic peak was observed at 59.9℃ (initial temperature). Under nitrogen protection, heating the sample to 100℃ and cooling to room temperature caused the crystal form E to transform into an amorphous form, as indicated by XRPD characterization. Figure 39 As shown. Meanwhile, the NMR results for crystal form E ( Figure 40 The results showed that 7.2 wt% IPAc was detected (API / IPAc molar ratio: 1:0.42). Based on the step-like weight loss observed in the TGA and the detection of IPAc solvent residue, crystal form E is presumed to be an IPAc solvate that becomes amorphous after desolventizing upon heating.

[0229] 5.6 Crystal form F

[0230] Crystal form F can be obtained by room temperature suspension and stirring. Crystal form F is obtained by suspending the starting sample thiophenepyrimidine derivative in EtOH / H2O (1:1, v / v) at room temperature and stirring. Its XRPD results are as follows: Figure 41 As shown. The TGA / DSC of this batch of samples ( Figure 42 The XRPD analysis showed that the sample lost 8.0% of its weight when heated to 80℃; endothermic peaks were observed at 79.0℃, 93.5℃, and 173.4℃ (peak temperatures). Under nitrogen protection, the sample of crystalline form F was heated to 75℃ and 98℃ and then cooled to room temperature, respectively. Crystalline form F transformed into amorphous (the sample at 75℃ contained a small amount of crystalline form F); after heating to 150℃ and cooling to room temperature, it transformed into crystalline form B, as shown in the XRPD characterization. Figure 43 As shown. 1 H NMR results ( Figure 44The results showed that the F-type sample contained 4.7 wt% residual EtOH (API / EtOH molar ratio: 1:0.60). Based on the step-like weight loss observed in the TGA and the detection of residual EtOH solvent, the F-type sample is presumed to be an EtOH solvate.

[0231] 5.7 Crystal form G

[0232] The crystal form G of this invention can be obtained by suspension stirring at room temperature or 50°C. Crystal form G is obtained by suspending and stirring the starting sample thiophene pyrimidine derivative in acetonitrile (ACN) / ethyl acetate (EtOAc) (1:1, v / v) at room temperature, and its XRPD results are as follows. Figure 45 As shown. The TGA / DSC data for this batch of crystal form G samples ( Figure 46 The results showed that the sample lost 1.7% of its weight when heated to 150°C; a sharp endothermic peak was observed at 144.4°C (initial temperature). 1 H NMR results ( Figure 47 The results showed that no ACN or EtOAc was detected in the G crystal form sample. Based on the small TGA weight loss and the absence of thermal changes observed before melting, crystal form G is presumed to be an amorphous form.

[0233] 5.8 Crystal form H

[0234] Crystal form H of this invention is obtained by suspending and stirring a mixture of non-crystalline A / B / C / G in EtOAc at room temperature. Its XRPD results are as follows: Figures 48-50 As shown. The TGA / DSC of this batch of H-type samples (…). Figure 51 The results showed a weight loss of 0.9% upon heating to 150°C; a sharp endothermic peak was observed at 167.5°C (initial temperature). Based on the relatively small TGA weight loss and the absence of thermal changes observed before melting, crystal form H is presumed to be an amorphous form.

[0235] 6. Study on thermodynamic stability relationship

[0236] To compare the thermodynamic stability relationships among different amorphous forms, a suspension competition test was conducted at room temperature and 50°C on four amorphous forms (crystal forms A / B / C / G). Four amorphous forms (crystal forms A / B / C / G) were found in the screening test; crystal form H was a new crystal form discovered in subsequent tests, therefore crystal form H was not added to the suspension stirring. ~2.5 mg of samples of each of the four different crystal forms (crystal forms A / B / C / G) were weighed into 1.5-mL vials, and pre-saturated solvents (EtOAc and IPA) were added to form a turbid solution. After approximately 3 days of magnetic stirring (~1000 rpm) at room temperature and 50°C, the solids were separated and XRPD was measured. The experimental results are summarized as follows: Figure 52 As shown, its XRPD characterization results ( Figure 53The results showed that crystal forms A / B / C / G transformed into a new crystal form, named crystal form H, after stirring in the EtOAc and IPA system (crystal form H was mixed with a small amount of crystal form G under IPA / room temperature conditions). The characterization results of crystal form H are shown in section 2.8, and it is presumed to be an amorphous form. These experimental results indicate that crystal form H is more stable than crystal forms A / B / C / G in the range of room temperature to 50°C.

[0237] 7. Polymorphism Evaluation

[0238] 7.1 Assessment of the physicochemical stability of the dominant crystal form (crystal form H)

[0239] In this invention, solid samples of crystal form H were placed openly at 25℃ / 60%RH for 1 week and at 40℃ / 75%RH for 1 week, respectively, and then sealed and placed at 80℃ for 36 hours. The changes in chemical purity and solid crystal form were evaluated using HPLC area purity and XRPD characterization results. The experimental results are summarized as follows: Figure 54 As shown, the HPLC area percentage purity / initial purity of thiophene pyrimidine derivative H after one week of storage under three conditions ranged from 99.0% to 100.1%, and the solid crystal form remained unchanged. Crystal form H exhibited good physicochemical stability at 25℃ and 40℃, but slight degradation was observed at 80℃. A summary of impurities detected by HPLC is shown below. Figure 55 As shown, the HPLC chromatogram is as follows: Figure 56 As shown, the XRPD overlay of solid crystal form H is as follows: Figure 57 As shown.

[0240] 7.2 Hygroscopicity assessment of the dominant crystal form (crystal form H)

[0241] The hygroscopicity of the dominant crystal form H was evaluated using dynamic moisture adsorption (DVS) testing. At a constant temperature of 25°C, the DVS test results ( Figure 58 The results showed that when the humidity increased from 0% to 80% RH, the sample of crystal form H gained 1.0 wt% due to moisture absorption, indicating that crystal form H has slight hygroscopicity. XRPD results ( Figure 59 The results show that the solid crystal form H is basically the same before and after DVS.

[0242] 7.3 Characterization of the morphology and size of the dominant crystal form (crystal form H)

[0243] This invention uses polarizing microscopy (PLM) to characterize and test crystal form H, such as... Figure 60 As shown, the H-type samples in this batch have small particles (0-10 μm) and agglomeration was observed.

[0244] 7.4 Equilibrium solubility of amorphous A / B / C / G / H in pure water

[0245] In this invention, five crystalline forms were magnetically stirred in pure water for 24 hours at room temperature, followed by centrifugation to separate the supernatant and the solid layer. The resulting supernatant was filtered through a 0.22 μm filter, and the solubility of the five crystalline forms A / B / C / G / H of the thiophene pyrimidine derivative was tested. The separated solids were then tested for changes in crystalline form using XRPD. The experimental results are summarized below. Figure 61 As shown, the mother liquor from the separated crystal forms B and H remained slightly turbid after filtration, presumably due to the small size of the solid particles, preventing the formation of a clear solution. XRPD characterization results are as follows. Figure 62 and Figure 63 As shown, after stirring and equilibrating the five crystal forms in pure water for 24 hours, the crystal forms remained unchanged.

[0246] 8. Preparation of the dominant crystal form (crystal form H)

[0247] 8.1 Repeat the preparation of crystal form H up to 15 mg

[0248] Starting with the receipt of amorphous thiophenepyrimidine derivatives, suspension and stirring were repeated in different solvents to prepare crystalline form H. ~15 mg of the starting sample was added to 0.5 mL of solvent to prepare a suspension. After magnetic stirring at room temperature and 50 °C for ~4 days, the solid was separated for XRPD testing. The experimental results are summarized as follows: Figure 64 As shown, starting with amorphous free base, crystalline form H was obtained after stirring in both IPA and EtOAc. A black colloidal solid was then obtained in dichloromethane (DCM). The XRPD results are shown below. Figure 65 As shown.

[0249] 8.2 Repeat the preparation of crystal form H up to 100 mg

[0250] ~100 mg of amorphous thiophenepyrimidine derivatives were added to 2 mL of solvent (IPA, EtOAc) to prepare a suspension. After magnetic stirring at room temperature for ~2 days, the solid was separated for XRPD testing, and crystal form H was successfully obtained in all cases. Experimental results and XRPD characterization are as follows: Figure 66 and Figure 67 As shown.

[0251] 9. Conclusion

[0252] This invention uses AN-002 thiophenepyrimidine derivatives as starting materials and employs various screening methods (including antisolvent addition, slow evaporation, slow cooling, room temperature / 50°C suspension stirring, gas-solid permeation, gas-liquid permeation, and polymer induction) to conduct a total of 101 polymorph screening experiments. Based on the characterization of the obtained solid samples and subsequent research evaluation, a total of 8 different crystal forms were discovered, including 5 amorphous forms A / B / C / G / H, 2 solvate forms E and F, and 1 hydrate form D (which could not be replicated). Heating experiments under nitrogen protection showed that crystal forms A / C / F all transformed into crystal form B upon heating; crystal form E transformed into an amorphous form upon heating. Suspension stirring results in different temperatures and solvent systems showed that amorphous form H was more stable than amorphous forms A / B / C / G at room temperature and 50°C.

[0253] In subsequent property evaluations, the physicochemical stability, hygroscopicity, and particle morphology of the dominant crystal form H were tested under different temperatures and humidity levels. Physicochemical stability tests showed that crystal form H exhibited relatively good physicochemical stability at 25℃ / 60%RH and 40℃ / 75%RH, with purity fluctuating between 99.9% and 100.1% after one week of storage. After 36 hours of storage at 80℃, the purity was 99.0% of the initial purity, indicating slight degradation. Dynamic moisture adsorption (DVS) tests showed that at a constant temperature of 25℃, the crystal form H sample gained 1.0% weight due to moisture absorption when humidity increased from 0%RH to 80%RH, indicating that crystal form H has slight hygroscopicity. Polarizing microscopy (PLM) tests revealed that the particles of this batch of crystal form H (806131-21-A1) were small (0–10 μm) and exhibited agglomeration.

[0254] Based on the above screening and evaluation, crystal form H is recommended as the crystal form of the advantageous thiophene pyrimidine derivative of this invention, that is, the crystal form of the thiophene pyrimidine derivative described in this invention for further development.

[0255] 10. Comparison of product data obtained by manufacturing enterprises (crystal form H obtained according to the preparation method of this invention):

[0256] The present invention commissioned a testing unit to test the crystal form (JFAN-1001) of the thiophene pyrimidine derivative prepared in this invention [corresponding to item 5.8 in the above examples]. The specific information is as follows:

[0257] Testing Unit: Kaihui Pharmaceutical (Shanghai) Co., Ltd.

[0258] Instrument model: Bruker D8 Advance;

[0259] Test conditions: Cu K

[0260] Operating voltage: 40kV;

[0261] Operating current: 40mA;

[0262] Starting angle: 4° (2-theta);

[0263] Termination angle: 40° (2-theta);

[0264] Interval: 0.05° / step;

[0265] Scanning speed: 0.5 seconds / step;

[0266] Test spectra and results: see Figure 68 .

[0267] The powder X-ray diffraction pattern of JFAN-1001 indicates that it is a crystalline powder. Upon comparison, its powder X-ray diffraction pattern is consistent with the stable crystal form results in the crystal form screening report; therefore, this product is in the H crystal form.

[0268] 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.

[0269] 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.

[0270] As used in this invention, the term "and / or" includes any one or more of the related listed items and all combinations thereof.

[0271] 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, characterized in that, The structural formula of the thiophene pyrimidine derivative is shown in formula (I):

2. The crystal form of the thiophene pyrimidine derivative as described in claim 1, characterized in that, The crystal forms of the thiophene pyrimidine derivatives are selected from the following group: crystal form A, crystal form B, crystal form C, crystal form D, crystal form E, crystal form F, crystal form G, and crystal form H.

3. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form H of the thiophenepyrimidine derivative has characteristic peaks at 2θ angles of 8.626, 10.658, 12.532, 19.409, 21.402, and 27.

008.

4. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The crystal form A has one or more characteristics selected from the group consisting of: 1) The crystal form A has a basic X-ray powder diffraction pattern as shown in Figure 24; 2) The crystal form A has a DSC spectrum as shown in Figure 25; 3) The crystal form A has a TGA spectrum as shown in Figure 25; 4) The crystal form A has XRPD images before and after heating, as shown in Figure 26; 5) The crystal form A has the basic characteristics shown in Figure 27. 1 H NMR spectrum; The DSC spectrum includes an endothermic peak at 172.5 °C.

5. The crystal form of the thiophenepyrimidine derivative as described in claim 2, characterized in that, The crystal form B has one or more characteristics selected from the group consisting of: 1) The crystal form B has a basic X-ray powder diffraction pattern as shown in Figure 28; 2) The crystal form B has a DSC spectrum as shown in Figure 29; 3) The crystal form B has a TGA spectrum as shown in Figure 29; 4) The crystal form B has the basic characteristics shown in Figure 30. 1 H NMR spectrum; The DSC spectrum includes an endothermic peak at 170.9 °C.

6. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The crystal form C has one or more characteristics selected from the group consisting of: 1) The crystal form C has a basic X-ray powder diffraction pattern as shown in Figure 31; 2) The crystal form C has a DSC spectrum as shown in Figure 32; 3) The crystal form C has a TGA spectrum as shown in Figure 32; 4) The crystal form C has XRPD images before and after heating, as shown in Figure 33; 5) The crystal form C has the basic characteristics shown in Figure 34. 1 H NMR spectrum; The DSC spectrum includes an endothermic peak at 172.6 °C.

7. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The crystal form D has one or more features selected from the group consisting of: 1) The crystal form D has a basic X-ray powder diffraction pattern as shown in Figure 35; 2) The crystal form D has a DSC spectrum as shown in Figure 36; 3) The crystal form D has a TGA spectrum as shown in Figure 36; The DSC spectrum includes an endothermic peak at 175.0 °C.

8. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The crystal form E has one or more characteristics selected from the group consisting of: 1) The crystal form E has a basic X-ray powder diffraction pattern as shown in Figure 37; 2) The crystal form E has a DSC spectrum as shown in Figure 38; 3) The crystal form E has a TGA spectrum as shown in Figure 38; 4) The crystal form E has the basic characteristics shown in Figure 39. 1 H NMR spectrum; 5) The crystal form E has XRPD images before and after heating, as shown in Figure 40; The DSC spectrum includes an endothermic peak at 79.6 °C.

9. The crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The crystal form F has one or more characteristics selected from the group consisting of: 1) The crystal form F has a basic X-ray powder diffraction pattern as shown in Figure 41; 2) The crystal form F has a DSC spectrum as shown in Figure 42; 3) The crystal form F has a TGA spectrum as shown in Figure 42; 4) The crystal form F has X-ray powder diffraction patterns before and after heating, as shown in Figure 43. 5) The crystal form F has the basic characteristics shown in Figure 44. 1 H NMR spectrum; The DSC spectrum includes an endothermic peak at 173.4 °C.

10. The crystal form of the thiophenepyrimidine derivative as described in claim 2, characterized in that, The crystal form G has one or more characteristics selected from the group consisting of: 1) The crystal form G has a basic X-ray powder diffraction pattern as shown in Figure 45; 2) The crystal form G has a DSC spectrum as shown in Figure 46; 3) The crystal form G has a TGA spectrum as shown in Figure 46; 4) The crystal form G has the basic characteristics shown in Figure 47. 1 H NMR spectrum; The DSC spectrum includes an endothermic peak at 149.8 °C.

11. The crystal form of the thiophenepyrimidine derivative as described in claim 2, characterized in that, The crystal form H has one or more characteristics selected from the group consisting of: 1) The crystal form H has a basic X-ray powder diffraction pattern as shown in Figure 49; 2) The crystal form H has a basic XRPD peak finding report as shown in Figure 50; 3) The crystal form H has a DSC spectrum as shown in Figure 51; 4) The crystal form H has a TGA spectrum as shown in Figure 51; The DSC spectrum includes an endothermic peak at 171.0 °C.

12. The crystal form of the thiophenepyrimidine derivative as described in claim 2, characterized in that, The crystal form H is an anhydrous crystal form with a single melting point and characteristic XRPD diffraction peaks; it has relatively good physicochemical stability at 25℃ / 60%RH and 40℃ / 75%RH; it has good thermal stability at 80℃; and it has low hygroscopicity at a constant temperature of 25℃ when the humidity increases from 0%RH to 80%RH.

13. A method for preparing the crystal form of the thiophene pyrimidine derivative as described in claim 2, characterized in that, The preparation method of the crystalline form H includes the following steps: using the received amorphous thiophene pyrimidine derivative as the starting material, preparing a suspension in a solvent, stirring at 20-55°C for 1-5 days, and then separating the solid to obtain the crystalline form H; and / or, The preparation method of the crystal form A includes the following steps: obtaining the crystal form A by adding n-heptane dropwise to an ethanol (EtOH) solution of a thiophene pyrimidine derivative of the starting sample for crystallization; and / or, The preparation method of the crystal form B includes one or more of the following methods: antisolvent addition, slow evaporation, slow cooling, gas-solid permeation, gas-liquid diffusion, and suspension stirring. Preferably, the preparation method includes the following steps: preparing the crystal form B by suspending and stirring the starting sample thiophene pyrimidine derivative in isopropanol (IPA) at room temperature; and / or, The preparation method of the crystal form C includes one or more of slow evaporation, gas-solid infiltration, and polymer-induced methods. Preferably, the preparation method includes the following steps: obtaining the crystal form C by slowly evaporating a methanol (MeOH) solution of a thiophene pyrimidine derivative of the starting sample at room temperature; and / or, The method for preparing crystal form D is a room-temperature suspension stirring method, comprising the following steps: obtaining crystal form D by suspending and stirring the starting sample thiophene pyrimidine derivative in IPA / H2O (aw~0.8) at room temperature; and / or, The method for preparing the crystal form E is a room-temperature suspension and stirring method, comprising the following steps: obtaining the crystal form E by suspending and stirring the starting sample thiophene pyrimidine derivative in MeOH / isopropyl ester (IPAc) (1:2, v / v) at room temperature; and / or, The method for preparing the crystal form F is a room-temperature suspension and stirring method, comprising the following steps: obtaining the crystal form F by suspending and stirring the starting sample thiophene pyrimidine derivative in EtOH / H2O (1:1, v / v) at room temperature; and / or, The preparation method of the crystal form G is a suspension stirring method at room temperature or 50°C. Preferably, the preparation method includes the following steps: obtaining the crystal form G by suspending and stirring the starting sample thiophene pyrimidine derivative in acetonitrile (ACN) / ethyl acetate (EtOAc) (1:1, v / v) at room temperature.

14. The preparation method according to claim 13, characterized in that, The solvent is selected from one or more of propanol, dichloromethane, and ethyl acetate; and / or, the stirring temperature is 20-55°C; and / or, the stirring time is 1-5 days; and / or, the ratio of the thiophene pyrimidine derivative to the solvent is (15-100) mg: (0.5-2) mL.

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

16. The use of the crystal form of the thiophene pyrimidine derivative as described in any one of claims 1-12, or the preparation method as described in claim 13 or 14, or the drug / drug composition as described in claim 15 in the preparation of a medicament for treating a disease caused by abnormal cell proliferation due to overexpression of EGFR.

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