A quinazoline compound, composition and use thereof

Abstract

The application provides a quinazoline compound, a composition and application thereof, and particularly relates to a compound shown in formula (I), a stereoisomer of the compound, and a pharmaceutically acceptable salt of the compound, a composition of the compound and application of the compound in preparation of a medicine as a tyrosine kinase inhibitor. The compound of the application has good inhibitory activity on EGFR and HER2 kinase.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a quinazoline compound, a composition, and its application. Background Technology

[0002] Epidermal growth factor receptor (ErbB) tyrosine kinases regulate cell proliferation, migration, differentiation, apoptosis, and cell movement through multiple pathways. In various forms of malignant tumors, ErbB family members and some of their ligands are often overexpressed, amplified, or mutated, making them important targets for cancer therapy. This family of protein kinases includes ErbB1 / EGFR / HER1, ErbB2 / HER2, ErbB3 / HER3, and ErbB4 / HER4. Among these, several kinase inhibitors based on EGFR and HER2 have been successfully developed for the treatment of non-small cell lung cancer and breast cancer. (Dienstmann R.,et.al.,(2001)PersonalizingTherapy with Targeted Agents in Non-Small Cell Lung Cancer.ONCOTARGET.2(3),165.;Mitri Z.,et.al.(2012)The HER2 Receptor in Breast Cancer: Pathophysiology,Clinical Use,and New Advances in Therapy.,Chemotherapy Research&Practice.,Volum 2012(23),743193.).

[0003] EGFR is widely expressed and plays a vital role in growth, development, and normal physiological functions. Furthermore, EGFR and its mediated signaling pathways also play a crucial role in tumorigenesis and development. However, EGFR expression is highly unstable, frequently exhibiting gene amplification and rearrangement, leading to alterations in the antigenic phenotype of tumor cells. The most common of these alterations is the epidermal growth factor receptor variant III (EGFRvIII).

[0004] EGFRvIII is a class of mutants of the epidermal growth factor receptor (EGFR) discovered in recent years that are expressed only on the surface of tumor cells, not normal tissue cells. Aberrant expression of EGFR is associated with the development of many malignant tumors, including glioma, small cell lung cancer, breast cancer, bladder cancer, and ovarian cancer.

[0005] Compared to the intact EGFR structure, exons 2-7 encoding the extracellular ligand-binding region of EGFRvIII are deleted, resulting in an 801-base pair loss. This causes exons 1 and 8 to connect, and a new glycine residue is created at this binding site, leading to the deletion of amino acids 6-273, thus losing the ability to bind to the ligand EGF. Without ligand binding, EGFRvIII induces unregulated structural activation of tyrosine kinases via dimerization and autophosphorylation, triggering downstream signaling and stimulating tumor cell proliferation.

[0006] Previous studies have shown that EGFRvIII can influence tumorigenesis and development by regulating multiple signaling pathways, including Ras / Raf / MEK / ERK, PI3 / AKT / mTOR, JAK / STAT, and PLC / PKC. EGFRvIII-positive tumor cells exhibit significantly increased tumorigenicity, primarily through inhibiting apoptosis, promoting tumor angiogenesis, and increasing invasiveness and migration, leading to uncontrolled spontaneous proliferation and metastasis of tumor cells. Furthermore, EGFRvIII plays a similar escape-like role during radiotherapy and chemotherapy for tumors.

[0007] Gliomas are common and highly aggressive malignant tumors, with glioblastoma (GBM) being the most malignant type. The effects of radiotherapy and chemotherapy are not ideal, and recurrence is common after surgery. Domestic and international studies have found that 40%–60% of GBMs significantly express EGFR, with the EGFRvIII mutant form being the predominant one. EGFRvIII, through receptor-independent autophosphorylation and tyrosine kinase activity, establishes a signaling pathway regulatory network, playing a crucial role in regulating GBM growth, metastasis, and angiogenesis.

[0008] Recent studies have shown that molecularly targeted therapies against EGFRvIII have demonstrated promising anti-tumor effects in both in vitro cell culture and in vivo animal models. Therefore, developing new molecularly targeted therapies against EGFRvIII will provide cancer patients, especially those with gliomas, with more effective and cost-effective treatment options, representing a significant unmet clinical need.

[0009] Drugs targeting EGFRvIII for the treatment of gliomas need to not only effectively cross the blood-brain barrier but also effectively inhibit EGFRvIII. Currently, there are no reports of compounds that can both cross the blood-brain barrier and inhibit EGFRvIII; therefore, research on EGFRvIII-driven gliomas has significant clinical value. Furthermore, most marketed EGFR and HER2 kinase inhibitors cannot cross the blood-brain barrier, and patients with EGFR-driven lung cancer and HER2-driven breast cancer generally have poor prognoses and a high risk of brain metastases. Currently, there are no approved effective drugs for the treatment of brain metastases; therefore, there is an urgent need to develop an EGFR inhibitor and / or HER2 inhibitor that can cross the blood-brain barrier. Summary of the Invention

[0010] In one aspect, this invention provides a compound of formula (I), a stereoisomer, and a pharmaceutically acceptable salt thereof, wherein,

[0011]

[0012] In equation (I), m is 0, 1, or 2;

[0013] R1 is hydrogen, a 4-7 membered heterocyclic group, or -NR. a R b ,

[0014] R a R b Each of these can be independently hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkyl substituted with hydroxyl, or C3-C6 alkyl substituted with C4. 1- C1-C6 alkyl groups substituted with C3 alkoxy groups, or C3-substituted alkyl groups... 3- C1-C6 alkyl substituted with C6 cycloalkyl,

[0015] The 4-7 membered heterocyclic group is a heterocyclic group containing 1-2 heteroatoms selected from N, O, or S. The heterocyclic group is either unsubstituted or substituted with C1-C3 alkyl, C1-C4 acyl, hydroxyl, cyano, aminoacyl, mono- or bis-C1-C3 alkyl-substituted aminoacyl, C 1- C3 alkyl sulfone group, C 1- One or both of the following substitutions are selected: C3 alkyl sulfoxide group, oxo (=O);

[0016] R2 is composed of 1 to 3 radicals selected from halogen, cyano, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 alkylthio, hydroxyl, C3-C4 cycloalkyl, or C 1- C1-C6 alkyl groups substituted or unsubstituted by substituents in C3 alkyl sulfone groups;

[0017] R3, R4, and R5 are each independently hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxy, or C3-C4 cycloalkyl, and at least one of them is halogen.

[0018] According to a preferred embodiment, m is 0 or 1.

[0019] R1 is a 4-7 member heterocyclic group or -NR a R b ,

[0020] R a R b Each of these can be independently hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkyl substituted with hydroxyl, or C6 alkyl substituted with C6 hydroxyl. 1- C3 alkoxy-substituted C1-C3 alkyl;

[0021] The 4-7 membered heterocyclic group is pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or thiomorpholinyl, and the above groups are not substituted or are substituted by one or two of methyl, ethyl, propyl, isopropyl, aldehyde, acetyl, propionyl, hydroxyl, cyano, aminoacyl, methyl sulfone, ethyl sulfone, propyl sulfone, isopropyl sulfone, methyl sulfoxide, ethyl sulfoxide, propyl sulfoxide, isopropyl sulfoxide, or oxo (=O).

[0022] R1 is 1-methylpyrrolidin-2-yl, 1-ethylpyrrolidin-2-yl, 1-propylpyrrolidin-2-yl, 1-isopropylpyrrolidin-2-yl, pyrrolidin-1-yl, piperidin-1-yl, 1-methylpiperazin-4-yl, 1-ethylpiperazin-4-yl, morpholino, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, thiomorpholino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, methylethylamino, methylpropylamino, methylamino, ethylamino, propylamino, isopropylamino, cyclopropylamino, cyclobutylamino, methylisopropylamino, N-methyl-N-cyclopropylamino, N-methyl-N-cyclobutylamino, or ethylpropylamino.

[0023] According to a preferred embodiment, R2 is a C1-C4 alkyl group substituted or unsubstituted by one to three substituents selected from fluorine, chlorine, cyano, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, methylthio, ethylthio, propylthio, isopropylthio, hydroxyl, cyclopropyl, cyclobutyl, methylsulfone, ethylsulfone, propylsulfone, or isopropylsulfone.

[0024] More preferably, R2 is methyl, ethyl, propyl, isopropyl, hydroxyethyl, hydroxypropyl, trifluoromethyl, fluoroethyl, fluoropropyl, 2,2,2-trifluoroethyl, 2,2-difluoroethyl, 3,3,3-trifluoropropyl, methoxyethyl, methoxypropyl, ethoxyethyl, ethoxypropyl, methylthioethyl, methylthiopropyl, ethylthioethyl, ethylthiopropyl, 2-hydroxy-2-methylpropyl, 3-hydroxy-3-methylbutyl, methylsulfonylpropyl, methylsulfonylethyl, ethylsulfonylethyl, ethylsulfonylpropyl, isopropylsulfonylethyl, isopropylsulfonylpropyl.

[0025] According to a preferred embodiment, R3, R4, and R5 are each independently hydrogen, fluorine, chlorine, and bromine, and at least one of them is fluorine, chlorine, or bromine.

[0026] More preferably, R3 and R5 are each independently hydrogen, fluorine, and chlorine, and R4 is chlorine.

[0027] Preferably, R3 is hydrogen, fluorine, or chlorine, R4 is chlorine, and R5 is fluorine.

[0028] The typical compounds involved in this application are as follows:

[0029]

[0030]

[0031] Another aspect of the present invention provides a pharmaceutical composition comprising the compound described in this application, a pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0032] The pharmaceutical composition of this application may also contain one or more other therapeutic agents.

[0033] The present invention also relates to a method for treating diseases or conditions mediated by kinases such as EGFR and HER2, comprising administering a therapeutically effective amount of the compound or a salt thereof described in this application to a patient (human or other mammal, especially human) in need, wherein the diseases or conditions mediated by kinases such as EGFR and HER2 include those mentioned above. Invention Details

[0035] Unless otherwise stated, the following terms used in this application (including the specification and claims) have the definitions given below. In this application, unless otherwise stated, the use of “or” or “and” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “having” is not restrictive. Section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter.

[0036] Unless otherwise specified, alkyl refers to a saturated straight-chain or branched hydrocarbon group having a specified number of carbon atoms. The terms C1-C6 alkyl refer to alkyl moieties containing 1 to 6 carbon atoms, and similarly, C1-C3 alkyl refer to alkyl moieties containing 1 to 3 carbon atoms. For example, C1-C6 alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, and 2-methylpentyl, etc.

[0037] When a substituent term such as "alkyl" is used in combination with other substituent terms, such as in the terms "C1-C3 alkoxy-C1-C6 alkylthio" or "hydroxy-substituted C1-C6 alkyl," the linking substituent term (e.g., alkyl or alkylthio) is intended to include a divalent portion, wherein the linking point is through the linking substituent. Examples of "C1-C3 alkoxy-C1-C6 alkylthio" include, but are not limited to, methoxymethylthio, methoxyethylthio, and ethoxypropylthio. Examples of "hydroxy-substituted C1-C6 alkyl" include, but are not limited to, hydroxymethyl, hydroxyethyl, and hydroxyisopropyl.

[0038] Alkoxy groups are alkyl-O- groups formed by a straight-chain or branched alkyl group and -O-, as previously described, such as methoxy, ethoxy, etc. Similarly, alkylthio groups are alkyl-S- groups formed by a straight-chain or branched alkyl group and -S-, as previously described, such as methylthio, ethylthio, etc.

[0039] Alkenyl and alkynyl groups include straight-chain, branched alkenyl or alkynyl groups. The terms C2-C6 alkenyl or C2-C6 alkynyl refer to a straight-chain or branched hydrocarbon group having at least one alkenyl or alkynyl group.

[0040] The term "halogenated alkyl," such as "halogenated C1-C6 alkyl," refers to a group having one or more halogen atoms, which may be the same or different, on one or more carbon atoms of an alkyl moiety comprising 1 to 6 carbon atoms. Examples of "halogenated C1-C6 alkyl" may include, but are not limited to, -CF3 (trifluoromethyl), -CCl3 (trichloromethyl), 1,1-difluoroethyl, 2,2,2-trifluoroethyl, and hexafluoroisopropyl. Similarly, the term "halogenated C1-C6 alkoxy" refers to a haloalkyl-O- group formed by the said haloated C1-C6 alkyl and -O-, which may be, for example, trifluoromethoxy, trichloromethoxy, etc.

[0041] The term "C1-C4 acyl" includes formyl (aldehyde) (-CHO), acetyl (CH3CO-), propionyl (C2H5CO-), etc. The term "amino acyl" refers to NH2CO-.

[0042] "Cycloalkyl" refers to a non-aromatic, saturated, cyclic hydrocarbon group containing a specified number of carbon atoms. For example, the term "(C3-C6)cycloalkyl" refers to a non-aromatic cyclic hydrocarbon ring having 3 to 6 carbon atoms. Exemplary "(C3-C6)cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0043] The term "aryl" refers to a group or portion comprising an aromatic monocyclic or bicyclic hydrocarbon group containing 6 to 12 carbon ring atoms and having at least one aromatic ring. Examples of "aryl" are phenyl, naphthyl, indenyl, and dihydroindenyl (indenyl). Typically, in the compounds of this invention, the aryl group is phenyl.

[0044] The term “heterocyclic group” as used herein, unless otherwise specified, refers to a stable, non-aromatic monocyclic saturated ring system consisting of a carbon atom and one to three heteroatoms selected from N, O, and S, wherein the N and S heteroatoms can be arbitrarily oxidized and the N heteroatoms can be arbitrarily quaternized. Examples of such heterocycles include, but are not limited to, azazolinyl, oxazolinyl, thiohexazolinyl, pyrrolyl, pyrrololinyl, pyrazolyl, pyrazolinyl, imidazolinyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, 1,3-dioxacyclopentyl, piperidinyl, piperazinyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiaranyl, 1,3-dioxyl, 1,4-dioxyl, 1,3-oxothiohexacyclopentyl, 1,3-oxothiohexacyclohexyl, 1,3-dithiaalkyl, 1,4-oxothiohexacyclopentyl, 1,4-oxothiohexacyclohexyl, 1,4-dithiaalkyl, morpholinyl, and thiomorpholinyl.

[0045] The term “carbonyl” refers to the -C(O)- group. The terms “halogen” and “halogen” refer to chlorine, fluorine, bromine, or iodine substituents. “Oxygen” refers to the oxygen moiety of a double bond; for example, if directly attached to a carbon atom to form a carbonyl moiety (C=O). “Hydroxy” is intended to represent the -OH group. The term “cyano” as used herein refers to the -CN group.

[0046] The term "independently" means that when more than one substituent is selected from many possible substituents, those substituents may be the same or different.

[0047] It is clear that compounds, isomers, crystal forms, or prodrugs of Formula I, and their pharmaceutically acceptable salts, can exist in solvated and unsolvated forms. For example, the solvated form can be a water-soluble form. This invention includes all such solvated and unsolvated forms.

[0048] In this application, the term "isomer" refers to different compounds having the same molecular formula, and may include various isomeric forms such as stereoisomers and tautomers. "Stereoisomers" are isomers that differ only in the spatial arrangement of atoms. Some compounds described herein contain one or more asymmetric centers, and therefore can produce enantiomers, diastereomers, and other stereoisomers that, according to absolute stereochemistry, can be defined as (R)- or (S)-. The chemical entities, pharmaceutical compositions, and methods of this invention are intended to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of the compounds can be analyzed by any suitable method, including but not limited to chiral chromatography and optical rotation determination, and the extent to which one stereoisomer outperforms the others can be determined.

[0049] Individual isomers (or mixtures enriched with isomers) of the present invention can be separated using methods known to those skilled in the art. For example, the separation can be performed as follows: (1) by forming diastereomer salts, complexes, or other derivatives; (2) by selective reaction with stereoisomer-specific reagents, such as by enzymatic oxidation or reduction; or (3) by gas-liquid chromatography or liquid chromatography in a chiral environment, such as on a chiral support (e.g., silica gel with chiral ligands) or in the presence of a chiral solvent. Those skilled in the art will understand that when a desired stereoisomer is converted into another chemical entity using one of the above separation methods, additional steps are required to release the desired form. Alternatively, specific stereoisomers can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by asymmetric conversion of one enantiomer into another.

[0050] When the compounds described herein contain olefin double bonds, unless otherwise stated, it means that the compounds include various cis-trans isomers.

[0051] "Tautomers" are structurally different isomers that can interconvert through tautomerization. Tautomerization is a form of isomerization and includes proton shift or proton transfer tautomerization, which can be considered a subset of acid-base chemistry. Proton shift or proton transfer tautomerization involves proton migration accompanied by bond order changes, often involving the exchange of a single bond with an adjacent double bond. When tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. An example of tautomerization is keto-enol tautomerization.

[0052] The compounds of this invention, which are active ingredients, and the methods for preparing these compounds are also part of this invention. Furthermore, some compounds can exist in polycrystalline form, and this form is also included in the present invention. Additionally, some compounds can form solvates with water (i.e., hydrates) or common organic solvents, and these solvates are also included within the scope of this invention.

[0053] The compounds of the present invention can be used for treatment in their free form or, where appropriate, in the form of pharmaceutically acceptable salts or other derivatives. As used herein, the term "pharmaceutically acceptable salt" refers to organic and inorganic salts of the compounds of the present invention that are suitable for human and lower animal use, without excessive toxicity, irritation, allergic reactions, etc., and have a reasonable benefit / risk ratio. Pharmaceutically acceptable salts of amines, carboxylic acids, phosphonates, and other types of compounds are well known in the art. These salts can be formed by reacting the compounds of the present invention with a suitable free base or acid. This includes, but is not limited to, salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, or by using methods well known in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucohepanoate, glycerophosphate, gluconate, hemisulfate, hexanoate, hydroiodate, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, dihydroxynaphthalate, pectate, persulfate, per-3-phenylpropionate, phosphate, picrate, propionate, stearate, sulfate, thiocyanate, p-toluenesulfonate, undecanoate, etc. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include suitable non-toxic ammonium, quaternary ammonium, and amino cations formed using halogen ions, hydroxide ions, carboxyl ions, sulfate ions, phosphate ions, nitrate ions, lower alkyl sulfonates, and aryl sulfonates.

[0054] Furthermore, the term "prodrug" as used herein refers to a compound that can be converted into the compound described in this invention in vivo. This conversion is influenced by the hydrolysis of the prodrug in the blood or its enzymatic conversion into the parent compound in the blood or tissues.

[0055] The pharmaceutical compositions of the present invention comprise additional active agents of the compounds described herein or pharmaceutically acceptable salts thereof, kinase inhibitors (small molecules, peptides, antibodies, etc.), immunosuppressants, anticancer drugs, antiviral agents, anti-inflammatory agents, antifungal agents, antibiotics, or antiangiogenic compounds; and any pharmaceutically acceptable carrier, adjuvant, or excipient.

[0056] The compounds of the present invention can be used alone or in combination with one or more other compounds of the present invention or with one or more other pharmaceutical agents. When administered in combination, the therapeutic agents can be formulated for simultaneous or sequential administration at different times, or the therapeutic agents can be administered as a single composition. The term "combination therapy" refers to the use of the compounds of the present invention in conjunction with another pharmaceutical agent, administered either simultaneously or sequentially, with the aim of achieving optimal drug efficacy. Co-administration includes simultaneous delivery formulations as well as separate formulations for each compound. Therefore, the administration of the compounds of the present invention can be used concurrently with other known therapies in the art, such as in cancer treatment using radiotherapy or adjunctive therapies such as cell growth inhibitors, cytotoxic agents, or other anticancer agents to improve cancer symptoms. The present invention is not limited to the order of administration; the compounds of the present invention can be administered prior to, concurrently with, or after other anticancer agents or cytotoxic agents.

[0057] To prepare the pharmaceutical ingredients of this invention, one or more compounds or salts of molecular formula (I) that are the active ingredients can be tightly mixed with a pharmaceutical carrier, as per conventional pharmaceutical formulation techniques, wherein the carrier can take a variety of forms depending on the preparation designed for different routes of administration (e.g., oral or parenteral). Suitable pharmaceutically acceptable carriers are well known in the art. Descriptions of some such pharmaceutically acceptable carriers can be found in the Handbook of Pharmaceutical Excipients, jointly published by the American Pharmaceutical Association and the British Pharmaceutical Society.

[0058] The pharmaceutical compositions of the present invention may be in the following forms, for example, suitable for oral administration, such as tablets, capsules, pills, powders, sustained-release forms, solutions, or suspensions; for parenteral injection, such as clear solutions, suspensions, or emulsions; or for topical application, such as ointments or creams; or as suppositories for rectal administration. The pharmaceutical ingredients may also be in unit dose form suitable for single-dose administration of precise dosage. The pharmaceutical ingredients will include a conventional pharmaceutical carrier or excipient and a compound prepared according to the present invention as the active ingredient; additionally, other medical or pharmaceutical preparations, carriers, excipients, etc., may also be included.

[0059] Therapeutic compounds can also be administered to mammals other than humans. The dosage given to a mammal will depend on the animal's species and its disease condition or disorder. Therapeutic compounds can be given to animals in the form of capsules, pills, tablets, or liquids. They can also be administered by injection or enema. We prepare these drug forms according to traditional methods that conform to veterinary practice standards. As an alternative, pharmaceutically synthesized drugs can be mixed with animal feed; therefore, concentrated feed additives or premixes can be prepared and mixed with ordinary animal feed.

[0060] Another object of the present invention is to provide a method for treating cancer in a subject in need, comprising administering to the subject a therapeutically effective amount of a composition containing the compound of the present invention.

[0061] This invention also includes the use of the compounds of this invention or pharmaceutically acceptable derivatives thereof in the preparation of medicaments for treating cancers and autoimmune diseases associated with tyrosine kinases EGFR and HER2. The cancers mentioned herein (including non-solid tumors, solid tumors, primary or metastatic cancers, as otherwise noted herein, and including cancers that are resistant or refractory to one or more other treatments) and other diseases (including, but not limited to, fundus diseases, psoriasis, atherosclerosis, pulmonary fibrosis, liver fibrosis, myelofibrosis, etc.) are also included in this invention. The cancers mentioned include, but are not limited to: non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, endometrial cancer, prostate cancer, bladder cancer, leukemia, gastric cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, chronic myeloid leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, nasopharyngeal carcinoma, esophageal cancer, brain tumor, B-cell and T-cell lymphoma, lymphoma, multiple myeloma, biliary carcinosarcoma, and bile duct carcinoma. Detailed Implementation

[0062] This invention also provides methods for preparing the corresponding compounds. The compounds described herein can be prepared using a variety of synthetic methods, including the methods described below. The compounds of this invention, or their pharmaceutically acceptable salts, isomers, or hydrates, can be synthesized using the methods described below, combined with synthetic methods known in the field of organic chemical synthesis, or by variations of these methods as understood by those skilled in the art. Preferred methods include, but are not limited to, the methods described below.

[0063] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all commercially available conventional products. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items. The embodiments provided below better illustrate the invention; unless otherwise specified, all temperatures are in °C. The naming of some compounds in this application is translated from ChemDraw nomenclature.

[0064] Synthesis of intermediates

[0065] Preparation of (R,E)-3-(1-methylpyrrolidone-2-yl)acryloyl chloride

[0066]

[0067] (R,E)-3-(1-methylpyrrolidone-2-yl)acrylic acid (160 mg, 1 mmol) was added to dry dichloromethane (3 ml), followed by oxalyl chloride (130 mg, 1 mmol) and DMF (1 drop, catalytic amount). The mixture was stirred at room temperature for 3 hours. The reaction system changed from turbid to clear. After concentration, an off-white solid was obtained.

[0068] Example 1. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methoxyquinazoline-6-yl)-4-(dimethylamino)but-2-eneamide

[0069]

[0070] Step 1): Synthesis of 4,5-dichloro-6-nitroquinazoline

[0071] Add 4.5 g (20 mmol) of 5-chloro-6-nitroquinazolin-4(3H)-one to 45 mL of thionyl chloride and 2 mL of DMF. Heat to 80 °C and reflux until the product is completely dissolved. Then react for another 2 hours, concentrate, add toluene and concentrate again to obtain 4.9 g of white solid product.

[0072] Step 2): 5-Chloro-N-(3-Chloro-2-fluorophenyl)-6-nitroquinazolin-4-amine

[0073] 4,5-Dichloro-6-nitroquinazoline (4.9 g, 20 mmol) was added to dry acetonitrile, and 3-chloro-2-fluoroaniline (4.35 g, 30 mmol) and triethylamine (3 g, 30 mmol) were added separately at 0°C. The mixture was heated to 50°C and reacted for 5 hours. After cooling and concentration, the product was washed with methanol to give 5.3 g of a white solid product, yield 75%; LC-MS: 353 [M+H] + ;

[0074] Step 3): N-(3-chloro-2-fluorophenyl)-5-methoxy-6-nitroquinazolin-4-amine

[0075] 5-Chloro-N-(3-chloro-2-fluorophenyl)-6-nitroquinazolin-4-amine (3.5 g, 10 mmol) was added to a mixture of DMF (15 mL) and sodium methoxide solution (15 mL of 30% sodium methoxide in methanol) at 0°C. The mixture was stirred for 2 hours, quenched with ice, filtered, and dried to give 4.1 g of a yellow solid product, yield 94%; MS: 349 [M+H]. + ;

[0076] Step 4): N 4 -(3-Chloro-2-fluorophenyl)-5-methoxyquinazolin-4,6-diamine

[0077] N-(3-chloro-2-fluorophenyl)-5-methoxy-6-nitroquinazolin-4-amine (1.75 g, 5 mmol) was added to ethanol, followed by the addition of iron powder and an aqueous solution of ammonium chloride. The mixture was heated to 50°C and reacted for 2 hours. After cooling and filtration, the product was washed with a large amount of dichloromethane. The filtrate was washed with brine, dried, and concentrated to give 1.6 g of a light purple solid product (98% yield). MS: 319 [M+H] + ;

[0078] Step 5): N 4 3-(3-chloro-2-fluorophenyl)-5-methoxyquinazoline-4,6-diamine (32 mg, 0.1 mmol) was added to NMP (1 mL) solution, and (E)-4-(dimethylamino)but-2-enoyl chloride (24 mg, 0.15 mmol) in dichloromethane (1 mL) was added at 0°C. The mixture was stirred for half an hour, quenched with water, and the pH was adjusted to 9 with sodium bicarbonate. The mixture was then extracted with dichloromethane, washed with saturated brine, dried and concentrated. The resulting oily substance was purified by column chromatography to give 16 mg of a white solid product. 1H NMR (400MHz, DMSO-d6) δ10.26(s,1H),10.03(s,1H),8.61–8.48(m,2H),8.36(d,J=9.0Hz,1H),7.61(d,J=9.1Hz,1H),7.39–7 .25(m,2H),6.82(dt,J=15.5,5.9Hz,1H),6.59(d,J=15.5Hz,1H),3.94(s,3H),3.12–3.05(m,2H),2.20(s,6H).MS:430[M+H] + .

[0079] Example 2. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methoxyquinazoline-6-yl)-4-(cyclopropyl(methyl)amino)but-2-enamide

[0080] The synthesis was carried out in the same manner as in Example 1, except that in step 5, (E)-4-(dimethylamino)but-2-enoyl chloride was used instead of (E)-4-(dimethylamino)but-2-enoyl chloride. 1 H NMR(400MHz,DMSO-d6)δ10.26(s,1H),10.02(s,1H),8.61–8.50(m,2H),8.37(d ,J=9.1Hz,1H),7.61(d,J=9.0Hz,1H),7.40–7.25(m,2H),6.86(dt,J=15.4,6.2 Hz,1H),6.56(d,J=15.4Hz,1H),3.94(s,3H),3.37–3.29(m,2H),2.29(s,3H),1 .76(tt,J=6.7,3.5Hz,1H),0.48–0.44(m,2H),0.39–0.31(m,2H).MS:456[M+H] + .

[0081]

[0082] Example 3. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2-methoxyethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-eneamide

[0083] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 2-methoxyethyl-1-ol in step 3. 1H NMR (400MHz, DMSO-d6) δ10.10(s,1H),9.84(s,1H),8.53(s,1H),8.43(d,J=9. 1Hz,1H),8.25–8.16(m,1H),7.63(d,J=9.0Hz,1H),7.42(s,1H),7.35–7.26(m ,1H),6.82(dt,J=15.4,5.8Hz,1H),6.45(d,J=15.5Hz,1H),4.23–4.16(m,2H) ,3.78–3.71(m,2H),3.20(s,3H),3.12–3.06(m,2H),2.20(s,6H).MS:474[M+H] + .

[0084] Example 4. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2-fluoroethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0085] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 2-fluoroethane-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ9.97 (d, J=20.0Hz, 2H), 8.58 (s, 1H), 8.39–8.26 (m, 2H), 7.65 (d, J= 9.1Hz,1H),7.39(ddd,J=8.4,6.8,1.7Hz,1H),7.30(td,J=8.1,1.4Hz,1H),6.81(dt,J=15.4 ,5.9Hz,1H),6.48(dt,J=15.4,1.6Hz,1H),4.91–4.84(m,1H),4.79–4.73(m,1H),4.38(dd,J =4.7,2.7Hz,1H),4.34–4.27(m,1H),3.09–3.07(d,J=5.9Hz,2H),2.19(s,6H).MS:462[M+H] + .

[0086]

[0087] Example 5. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2-hydroxyethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0088] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 2-hydroxyethyl-1-ol in step 3. 1H NMR(400MHz,DMSO-d6)δ10.22(s,1H),10.05(s,1H),8.54-8.51(m,2H),8.17(dd ,J=8.4,7.1Hz,1H),7.62(d,J=9.1Hz,1H),7.41(dd,J=8.3,6.8Hz,1H),7.29(d,J =8.2Hz,1H),6.82(dt,J=15.4,6.0Hz,1H),6.43(d,J=15.4Hz,1H),5.75(s,1H), 4.15-4.13(m,2H),3.85(s,2H),3.08(d,J=6.0Hz,2H),2.19(s,6H).MS:460[M+H] + .

[0089] Example 6. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3-methoxypropoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0090] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 3-methoxypropyl-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.12(s,1H),9.90(s,1H),8.59(s,1H),8.43(dd,J=8.7,7.3 Hz,1H),8.24(d,J=9.0Hz,1H),7.63(d,J=9.0Hz,1H),7.42-7.29(m,2H),6.82(dt,J=1 5.4,5.9Hz,1H),6.48(d,J=15.5Hz,1H),4.11(t,J=6.4Hz,2H),3.50(t,J=6.1Hz,2H) ,3.14(s,3H),3.09(d,J=6.0Hz,2H),2.20(s,6H),2.09(q,J=6.3Hz,2H).MS:488[M+H] + .

[0091]

[0092] Example 7. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3-(methylsulfonyl)propoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0093] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 3-methylsulfonylpropyl-1-ol in step 3. 1H NMR (400MHz, DMSO-d6) δ10.00(s,1H),9.95(s,1H),8.60(s,1H),8.45(dd,J=8.6,7. 3Hz,1H),8.24(d,J=9.0Hz,1H),7.65(d,J=9.0Hz,1H),7.42-7.29(m,2H),6.82(dt, J=15.4,5.9Hz,1H),6.51(d,J=15.7Hz,1H),4.13(t,J=6.7Hz,2H),3.34–3.26(m,2H ),3.09(d,J=5.9Hz,2H),2.96(s,3H),2.34–2.26(m,2H),2.20(s,6H).MS:536[M+H] + .

[0094] Example 8. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methoxyquinazoline-6-yl)-4-(isopropyl(methyl)amino)but-2-enoyl amide was synthesized by the same method as in Example 1, except that in step 5, (E)-4-(dimethylamino)but-2-enoyl chloride was used instead of (E)-4-(dimethylamino)but-2-enoyl chloride. 1 H NMR (400MHz, DMSO-d6) δ10.27(s,1H),10.04(s,1H),8.62–8.49(m,2H),8.36(d,J=9.1Hz,1H),7.62(d,J=9.1Hz,1H),7.39-7.29(m,2H),6.82(dt,J =15.4,5.7Hz,1H),6.60(d,J=15.4Hz,1H),3.94(s,3H),3.20(d,J=5.7Hz ,2H),2.86-2.80(m,1H),2.15(s,3H),0.99(d,J=6.5Hz,6H).MS:458[M+H] + .

[0095]

[0096] Example 9. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methoxyquinazoline-6-yl)-4-(isopropylamino)but-2-enamide

[0097] The synthesis was carried out in the same manner as in Example 1, except that in step 5, (E)-4-(dimethylamino)but-2-enoyl chloride was used instead of (E)-4-(isopropylamino)but-2-enoyl chloride for the reaction. 1H NMR (400MHz, DMSO-d6) δ10.28(s,1H),10.04(s,1H),8.60(s,1H),8.57–8. 51(m,1H),8.35(d,J=9.1Hz,1H),7.63(d,J=9.1Hz,1H),7.42–7.26(m,2H), 6.91(dt,J=15.4,5.4Hz,1H),6.60(dt,J=15.3,1.8Hz,1H),3.94(s,3H),3. 41(d,J=5.5Hz,2H),2.84-2.78(m,1H),1.04(d,J=6.2Hz,6H).MS:444[M+H] + .

[0098] Example 10. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-isopropoxyquinazoline-6-yl)-4-(dimethylamino)but-2-eneamide

[0099] The synthesis was carried out using the same method as in Example 1, except that sodium isopropoxide was used instead of sodium methoxide in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.36 (s, 1H), 10.04 (s, 1H), 8.69 (t, J = 7.6Hz, 1H), 8.63(s,1H),8.14(d,J=9.0Hz,1H),7.62(d,J=9.0Hz,1H),7.39–7.25(m,2H ),6.82(dt,J=15.5,5.9Hz,1H),6.49(d,J=15.5Hz,1H),4.45(p,J=6.3Hz,1 H),3.08(d,J=6.0Hz,2H),2.19(s,6H),1.32(d,J=6.2Hz,6H).MS:458[M+H] + .

[0100]

[0101] Example 11. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3-hydroxypropoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0102] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 3-hydroxypropyl-1-ol in step 3. 1H NMR(400MHz,DMSO-d6)δ10.19(s,1H),9.98(s,1H),8.58(s,1H),8.54–8.39 (m,2H),7.63(d,J=9.1Hz,1H),7.42–7.25(m,2H),6.82(dt,J=15.4,6.0Hz,1 H),6.55–6.46(m,1H),4.98(s,1H),4.15(t,J=6.4Hz,2H),3.65(t,J=6.0Hz, 2H),3.10(d,J=6.1Hz,2H),2.20(s,6H),2.01(q,J=6.1Hz,2H).MS:474[M+H] + .

[0103] Example 12. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2-(methylthio)ethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide was synthesized in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 2-methylthioethyl-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.05(s,1H),9.94(s,1H),8.55(s,1H),8.30(d,J=9.1Hz,1H ),8.22(t,J=7.7Hz,1H),7.63(d,J=9.0Hz,1H),7.42(t,J=7.5Hz,1H),7.30(t,J=8.2 Hz,1H),6.83(dt,J=15.5,5.9Hz,1H),6.50(d,J=15.4Hz,1H),4.19(t,J=6.4Hz,2H), 3.09(d,J=5.8Hz,2H),2.98(t,J=6.3Hz,2H),2.19(s,6H),1.97(s,3H).MS:490[M+H] + .

[0104]

[0105] Example 13. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2-hydroxy-2-methylpropoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0106] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 2-hydroxy-2-methylprop-1-ol in step 3. 1H NMR(400MHz,DMSO-d6)δ10.12(s,2H),8.50(s,1H),8.35(d,J=9.0Hz,1H),8.0 1(t,J=7.5Hz,1H),7.62(d,J=9.0Hz,1H),7.44(dd,J=8.3,6.8Hz,1H),7.29(t, J=8.1Hz,1H),6.81(dt,J=15.4,6.1Hz,1H),6.33(d,J=15.4Hz,1H),5.66(s,1H ),3.87(s,2H),3.08(dd,J=6.1Hz,2H),2.18(s,6H),1.26(s,6H).MS:488[M+H] + .

[0107] Example 14. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3-hydroxy-3-methylbutoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0108] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 3-hydroxy-3-methylbut-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.25(s,2H),8.69–8.14(m,3H),7.58(s,1H),7.35–7.25(m,2H),6.81(dt,J=15.4,6.0Hz,1H),6.54( d,J=15.4Hz,1H),4.18(t,J=6.7Hz,2H),3.08(d,J=6.1Hz,2H),2.19(s,6H),2.00(t,J=6.8Hz,2H),1.13(s,6H).MS:502[M+H] + .

[0109]

[0110] Example 15. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3-fluoropropoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0111] The synthesis was carried out using the same method as in Example 1, except that sodium methoxide was replaced with sodium 3-fluoropropyl-1-ol in step 3. 1H NMR(400MHz,DMSO-d6)δ10.07(s,1H),9.95(s,1H),8.60(s,1H),8.54–8.37(m,1H),8.19(d, J=9.0Hz,1H),7.64(d,J=9.0Hz,1H),7.39(dd,J=8.4,6.8Hz,1H),7.31(t,J=8.2Hz,1H),6.81 (dt,J=15.5,5.9Hz,1H),6.47(d,J=15.5Hz,1H),4.72(t,J=5.8Hz,1H),4.60(t,J=5.8Hz,1H) ,4.14(t,J=6.5Hz,2H),3.08(dd,J=5.9Hz,2H),2.31–2.15(m,2H),2.19(s,6H).MS:476[M+H] + .

[0112] Example 16. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2,2,2-trifluoroethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0113] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 2,2,2-trifluoroethyl-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.04(s,1H),9.59(s,1H),8.61(d,J=8.0Hz,1H), 8.39(s,1H),8.14(d,J=9.0Hz,1H),7.69(d,J=9.0Hz,1H),7.43–7.35(m,1H ),7.31(t,J=8.2Hz,1H),6.81(dt,J=15.5,5.9Hz,1H),6.43(d,J=15.5Hz,1 H),4.77(q,J=9.0Hz,2H),3.09(d,J=6.0Hz,2H),2.20(s,6H).MS:498[M+H] + .

[0114]

[0115] Example 17. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(2,2-difluoroethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0116] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 2,2-difluoroethyl-1-ol in step 3.1 H NMR(400MHz,DMSO-d6)δ10.01(s,1H),9.74(s,1H),8.58(s,1H),8.33–8.2 4(m,1H),8.19(d,J=9.0Hz,1H),7.66(d,J=9.0Hz,1H),7.42(dd,J=8.4,6.7 Hz,1H),7.31(t,J=8.2Hz,1H),6.82(dt,J=15.4,5.9Hz,1H),6.63–6.30(m ,2H),4.38(t,J=15.4Hz,2H),3.12–3.06(m,2H),2.20(s,6H).MS:480[M+H] + .

[0117] Example 18. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-(3,3,3-trifluoropropoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-eneamide

[0118] The synthesis was carried out in the same manner as in Example 1, except that sodium methoxide was replaced with sodium 3,3,3-trifluoropropyl-1-ol in step 3. 1 H NMR (400MHz, DMSO-d6) δ9.93(s,1H),9.73(s,1H),8.55(s,1H),8.17(s,2H),7.64(d,J=9.2Hz,1H),7.43(t,J=7.5Hz,1H),7.30(t,J=8.4Hz, 1H),6.81(d,J=15.3Hz,1H),6.48(d,J=15.4Hz,1H),4.24(t,J=6.4Hz,2H),3.11–3.05(m,2H),3.04–2.96(m,2H),2.19(s,6H).MS:512[M+H] + .

[0119]

[0120] Example 19. (E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-methoxyquinazolin-6-yl)-4-(dimethylamino)but-2-eneamide

[0121] The synthesis was carried out using the same method as in Example 1, except that in step 2, 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline for the reaction. 1H NMR (400MHz, DMSO-d6) δ10.27(s,1H),10.06(s,1H),8.63–8.53(m,2H),8.38(d,J=9.1Hz,1H),7.62(t,J=8.7Hz,2H) ,6.82(dt,J=15.5,5.8Hz,1H),6.59(d,J=15.4Hz,1H),3.93(s,3H),3.10(d,J=5.7Hz,2H),2.21(s,6H).MS:464[M+H] + .

[0122] Example 20. (R,E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-methoxyquinazoline-6-yl)-3-(1-methylpyrrolidone-2-yl)acrylamide

[0123] The synthesis was carried out in the same manner as in Example 1, except that 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline in step 2, and (R,E)-3-(1-methylpyrrolidin-2-yl)acryloyl chloride was used instead of (E)-4-(dimethylamino)but-2-enoyl chloride in step 5. 1 H NMR (400MHz, DMSO-d6) δ10.27(s,1H),10.04(s,1H),8.63–8.53(m,2H),8.39(d,J=9.1H z,1H),7.67–7.58(m,2H),6.71(dd,J=15.3,7.5Hz,1H),6.57(d,J=15.3Hz,1H),3.94(s, 3H),3.04(dd,J=9.8,7.1Hz,1H),2.76(q,J=7.9Hz,1H),2.22(s,3H),2.23–2.13(m,1H) ,2.02(dtd,J=12.3,8.4,5.9Hz,1H),1.82–1.68(m,2H),1.66–1.52(m,1H).MS:490[M+H] + .

[0124]

[0125]

[0126] Example 21. (E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-(2-hydroxyethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0127] The synthesis was carried out in the same manner as in Example 1, except that 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline in step 2, and sodium 2-hydroxyethyl-1-ol was used instead of sodium methoxide in step 3. 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),10.05(s,1H),8.58–8.49(m,2H),8.23(t,J=8.5Hz,1H),7.67–7.56(m,2H),6.82(dt,J=15.4,6.0Hz,1 H),6.43(dt,J=15.4Hz,1H),5.75–5.68(m,1H),4.15-4.13(m,2H),3.84(q,J=4.3Hz,2H),3.10(dd,J=6.0Hz,2H),2.20(s,6H).MS:494[M+H] + .

[0128] Example 22. (R,E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-(2-hydroxyethoxy)quinazolin-6-yl)-3-(1-methylpyrrolidone-2-yl)acrylamide

[0129] The synthesis was carried out using the same method as in Example 1, except that 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline in step 2, sodium 2-hydroxyethyl-1-ol was used instead of sodium methoxide in step 3, and (R,E)-3-(1-methylpyrrolidone-2-yl)acryloyl chloride was used instead of (E)-4-(dimethylamino)but-2-enoyl chloride in step 5. 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),10.04(s,1H),8.54(d,J=9.6Hz,2H),8.24(t,J=8.5Hz,1H),7. 67–7.56(m,2H),6.71(dd,J=15.3,7.8Hz,1H),6.41(d,J=15.3Hz,1H),5.73(t,J=4.3Hz,1H),4.14(d d,J=5.2,3.3Hz,2H),3.85(q,J=4.3Hz,2H),3.04(dd,J=9.8,7.3Hz,1H),2.76(q,J=8.0Hz,1H),2.21 (s,3H),2.18(t,J=8.8Hz,1H),2.08–1.94(m,1H),1.82–1.68(m,2H),1.66–1.52(m,1H).MS:520[M+H] + .

[0130]

[0131] Example 23. (E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-(2-methoxyethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide

[0132] The synthesis was carried out in the same manner as in Example 1, except that 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline in step 2, and sodium 2-methoxyethyl-1-ol was used instead of sodium methoxide in step 3. 1 H NMR (400MHz, DMSO-d6) δ9.85(br,2H),8.43(d,J=9.1Hz,1H),8.37(s,1H),8.08(s,1H),7.51(d,J=9.0Hz,2H),6.80(dt,J=15.4,5.8Hz, 1H),6.39(d,J=15.4Hz,1H),4.26–4.19(m,2H),3.75–3.68(m,2H),3.25(s,3H),3.08(dd,J=5.8,1.7Hz,2H),2.19(s,6H).MS:508[M+H] + .

[0133] Example 24. (R,E)-N-(4-((3,4-dichloro-2-fluorophenyl)amino)-5-(2-methoxyethoxy)quinazolin-6-yl)-3-(1-methylpyrrolidone-2-yl)acrylamide

[0134]

[0135] The synthesis was carried out using the same method as in Example 1, except that 3,4-dichloro-2-fluoroaniline was used instead of 3-chloro-2-fluoroaniline in step 2, sodium 2-methoxyethyl-1-ol was used instead of sodium methoxide in step 3, and (R,E)-3-(1-methylpyrrolidone-2-yl)acryloyl chloride was used instead of (E)-4-(dimethylamino)but-2-enoyl chloride in step 5. 1HNMR (400MHz, DMSO-d6) δ10.11(s,1H),9.84(s,1H),8.54(s,1H),8.45(d,J=9.1Hz,1H),8.29(t,J= 8.5Hz,1H),7.67–7.56(m,2H),6.71(dd,J=15.3,7.6Hz,1H),6.43(d,J=15.3Hz,1H),4.23–4.16(m, 2H),3.74(d,J=6.0Hz,2H),3.20(s,3H),3.04(dd,J=9.7,7.0Hz,1H),2.78(q,J=7.9Hz,1H),2.22(s ,3H),2.19(q,J=8.8Hz,1H),2.08–1.95(m,1H),1.81–1.68(m,2H),1.65–1.51(m,1H).MS:534[M+H] + .

[0136] Experimental Example 1. Small molecule compounds inhibit EGFR WT and HER2 kinase activity test

[0137] Reagents and consumables: ULight™-labeled Poly GT Peptide (Perkin Elmer, catalog number TRF-0100-M); ULight™-labeled JAK-1 (Try1023) Peptide (Perkin Elmer, catalog number TRF-0121-M); Eu-W1024-labeled Anti-Phosphotyrosine Antibody (PT66) (Perkin Elmer, catalog number AD0068); 10× Detection Buffer (Perkin Elmer, catalog number CR97-100); HER2 kinase (CarnaBiosciences, catalog number 08-016); EGFR kinase (CarnaBiosciences). Biosciences (catalog number 08-115); HEPES (GIBCO, catalog number 15630-080); EGTA (Sigma, catalog number 03777-10G); EDTA (Sigma, catalog number EDS-100G); MgCl2 (Sigma, catalog number 63069-100ML); DTT (Sigma, catalog number 43816-10ML); Tween-20 (Sigma, catalog number P7949-100ML); DMSO (Life Science, catalog number 0231-500ML); 384-well plate (PerkinElmer, catalog number 607290); Multifunctional plate reader (PerkinElmer, catalog number Envision)

[0138] Compound solution preparation: Dissolve the test compound in DMSO to prepare a 10 mM stock solution. Before use, dilute the compound in DMSO to 0.25 mM (100-fold final concentration dilution), and perform 3-fold serial dilutions, for a total of 11 serial dilutions. When adding the drug, dilute with buffer to a 4-fold final concentration.

[0139] HER2 kinase assay: Prepare buffer solutions, including 40 nM 4X HER2 kinase solution, 40 μM 4X ATP solution, and 400 nM 4× ULight solution. TM -Labeled Ploy GT Peptide substrate solution. After preparation, mix the enzyme with pre-diluted compounds of different concentrations, incubate at room temperature for 5 minutes, and set up replicates for each concentration. Add the corresponding substrate and ATP, and react at room temperature for 120 minutes (including positive and negative controls). After the reaction, add PT66 detection antibody, incubate at room temperature for 60 minutes, and then detect with Envision.

[0140] EGFR WTKinase assay: Prepare buffer solutions, including 3.48 nM 4X EGFR kinase solution, 600 μM 4X ATP solution, and 400 nM 4× ULight solution. TM -labeled JAK-1(Try1023) Peptide substrate solution. After preparation, the enzyme was mixed with pre-diluted compounds of different concentrations and incubated at room temperature for 5 minutes, with replicates for each concentration. The corresponding substrate and ATP were added, and the reaction was carried out at room temperature for 120 minutes (including positive and negative controls). After the reaction, PT66 detection antibody was added, and the mixture was incubated at room temperature for 60 minutes before detection using Envision.

[0141] Data calculation: Well readings and inhibition rates were calculated using an Excel spreadsheet. Well reading = 10000 * (well EU665 value) / (well EU615 value), inhibition rate = [(positive control well reading - experimental well reading) / (positive control well reading - negative control well reading)] * 100%. The compound concentration and corresponding inhibition rate were then input into GraphPad Prism for IC50 calculation. 50 value.

[0142] Table 1 shows that the compounds in this application can inhibit EGFR. WT The activity of HER2 tyrosine kinase was measured, with some compounds exhibiting particularly strong inhibitory effects. The test results are summarized in Table 1 below.

[0143] Table 1 lists the effects of some compounds from this application on EGFR. WT The results of the assay for HER2 tyrosine kinase inhibitory activity, where A represents IC50. 50 Less than or equal to 1 nM, B represents IC 50 For values ​​greater than 1 nM but less than or equal to 10 nM, C represents IC. 50 Greater than 10 nM but less than or equal to 100 nM, D represents IC 50 For values ​​greater than 100 nM but less than or equal to 1000 nM, NT indicates no relevant results.

[0144] Table 1. Results of the assay of inhibitory activity of the compounds of the present invention against EGFR and HER2 kinases.

[0145]

[0146] As can be seen from the results in Table 1 above, the compounds of this application exhibit good to excellent inhibitory activity against both HER2 and EGFR kinases. Among them, they exhibit very strong inhibitory activity against EGFR, thus showing a certain degree of selectivity relative to HER2 kinase.

[0147] Experimental Example 2. Test of the Inhibition of Cell Proliferation by Small Molecule Compounds

[0148] This application used the CTG method to detect the in vitro antiproliferative activity of the compounds of the present invention against HCC-827, Ba / F3-EGFR-VIII and Ba / F3 EGFR D770_N771insSVD cell lines cultured in vitro.

[0149] Reagents and consumables: RPMI 1640 (ThermoFisher, catalog number C11875500BT); DMEM (ThermoFisher, C11995500BT); Fetal bovine serum (Hyclone, catalog number SV30087.03); 0.25% trypsin-EDTA (ThermoFisher, catalog number 25200072); Penicillin-streptomycin (Hyclone, catalog number SV30010); DSMO (Life Science, catalog number 0231-500ML); CTG test kit (Promega, catalog number G9243); 96-well plate (Corning, catalog number 3599); Multifunctional plate reader (Perkin Elmer, catalog number Envision).

[0150] Cell lines: HCC-827 (from ATCC), Ba / F3-EGFR-VIII and Ba / F3 EGFR D770_N771insSVD (all from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd.); all the above cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. HCC-827 was cultured in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin.

[0151] Specific experimental methods:

[0152] 1. Dissolve the test compound in DSMO to form a stock solution and perform serial dilutions, then dilute with the appropriate culture medium to obtain a working concentration solution at 5 times.

[0153] 2. Dilute cells in the logarithmic growth phase with culture medium to a specific cell density, and add 80 μL of cell suspension to each well of a 96-well plate, ensuring a cell seeding density of 3000 cells / well for HCC-827, Ba / F3-EGFR-VIII, and Ba / F3 EGFR D770_N771insSVD. Ba / F3-EGFR-VIII and Ba / F3 EGFR D770_N771insSVD cells proceed directly to the next step of compound treatment, while HCC-827 cells require overnight incubation at 37°C with 5% CO2 gas for cell adhesion before compound treatment.

[0154] 3. Add 20 μL of the compound solution to each well of a 96-well plate containing pre-seeded cells. The highest concentration of the compound to be tested was 10 μM, with a total of 9 concentrations, serially diluted 4-fold, in duplicate. A control group without the compound was also included.

[0155] 4. After culturing cells for another 72 hours, cell viability was assessed using a CTG assay kit. Signal values ​​were read using a Perkin Elmer multi-function plate reader, and dose-response curves were generated and IC50 calculated using GraphPad Prism software. 50 .

[0156] Table 2 lists the results of antiproliferative activity assays of representative compounds of this invention against HCC-827, Ba / F3-EGFR-VIII, and Ba / F3 EGFRD770_N771insSVD cells. Where A represents IC50. 50 Less than or equal to 5nM, B represents IC 50 Greater than 5 nM but less than or equal to 50 nM, C represents IC 50 Greater than 50 nM but less than or equal to 500 nM, D represents IC 50 For values ​​greater than 500 nM but less than or equal to 5000 nM, NT indicates no relevant results.

[0157] Table 2. Results of antiproliferative activity assays of representative compounds of the present invention against HCC-827, Ba / F3-EGFR-VIII, and Ba / F3 EGFR D770_N771insSVD cells.

[0158]

[0159] Table 2 shows that the compounds of this application exhibit certain antitumor proliferative activity against all the cell lines tested above. In particular, the compounds of this application showed excellent activity against the HCC-827 and Ba / F3-EGFR-VIII cell lines.

[0160] Experimental Example 3. Pharmacokinetic Study of Small Molecule Compounds

[0161] This study investigated the pharmacokinetic characteristics of the compound and its ability to penetrate the blood-brain barrier by administering a portion of the compound to SD rats via single oral and intravenous injection.

[0162] (a) Reagents, instruments and animals used

[0163] Table 3. Test Reagents

[0164]

[0165] Table 4. Test Instruments

[0166]

[0167] Table 5. Experimental Rats

[0168]

[0169] (II) Sample Preparation

[0170] 1. Intravenous (IV) group: Weigh an appropriate amount of the test compound and completely dissolve it in an appropriate volume of solvent (DMSO / Solutol / H2O = 5 / 10 / 85V / V, with 2 moles of HCl added). Stir, vortex, and / or sonicate the solution. After obtaining the solution, gradually increase the solvent to the final volume to achieve the target concentration, vortex, and sonicate to obtain a homogeneous solution, which is then filtered through a 0.22 μm PVDF membrane.

[0171] 2. Oral (PO) group: Weigh an appropriate amount of the test compound and completely dissolve it in an appropriate volume of solvent (DMSO / Solutol / H2O = 5 / 10 / 85V / V, with 2 moles of HCl added). Stir, vortex, and / or sonicate the solution. After obtaining the solution, gradually increase the solvent to the final volume to achieve the target concentration, then vortex and sonicate to obtain a homogeneous solution.

[0172] (III) Administration and Sampling of Rats

[0173] Animals were randomly assigned to groups based on their body weight, with each group having a similar body weight (not exceeding ±20% of the average body weight). Group IV was fasted, while group PO was fasted overnight (>12 hours) and given food 2 hours after administration. All animals had free access to water. Tables 6 and 7 below provide the administration and pharmacokinetic sampling protocols, respectively.

[0174] Table 6. Dosing Regimen

[0175]

[0176] Table 7. Pharmacokinetic Sampling Protocol

[0177]

[0178] Rats were administered the drug according to the above protocol, and blood and brain tissue samples were collected and processed at predetermined time points (collection and processing were performed according to conventional methods in the art).

[0179] (iv) Sample Analysis

[0180] The brain was weighed and homogenized with 4 times the volume of homogenate (acetonitrile / water = 1:1). The whole blood sample and brain homogenate were each added with 6 times the volume of acetonitrile, vortexed for 1 min, and then centrifuged at 4500 rpm for 15 min at 4°C. The supernatant was diluted 2 times with ultrapure water, and the samples were analyzed by LC / MS.

[0181] (V) Data Analysis:

[0182] Pharmacokinetic parameters will be calculated using WinNonlin software. If applicable plasma drug concentration-time data are available, the following pharmacokinetic parameters will be calculated: CL (clearance); Vp d (Apparent volume of distribution); T 1 / 2 (Eliminating half-life); C max (Peak concentration); T max (Time to peak concentration); AUC (Area under the plasma concentration-time curve); MRT (Mean residence time); F% (Bioavailability).

[0183] The test results are shown in Tables 8-10 below, which present the rat blood drug concentrations of Compound 1 from the embodiments of this application at various time points, as well as the values ​​of various pharmacokinetic parameters. The concentrations and ratios of Compound 1 in the rat brain and blood are also given. The results show that Compound 1 of this application exhibits excellent ability to penetrate the blood-brain barrier. This indicates that the compound not only possesses excellent EGFR kinase inhibitory activity and can inhibit cell proliferation at the cellular level, but also has excellent ability to penetrate the blood-brain barrier, making it a promising candidate for applications in EGFR kinase-mediated diseases, especially those related to brain metastases.

[0184] Table 8. Rat blood drug concentrations of compound 1 from the embodiments of this application.

[0185]

[0186] Table 9. Rat pharmacokinetic parameters of the compounds of Example 1 of this application

[0187]

[0188] Table 10. Concentrations and ratios of compound 1 in brain and whole blood in the embodiments of this application (PO 10 mg / kg, sampling time, 2 h after administration)

[0189] Example Blood drug concentration (ng / mL) Brain concentration (ng / g) Brain / blood ratio 1 480 1468 3.06

[0190] The above description represents the preferred embodiments of the present invention. It should be noted that, for those skilled in the art, various improvements and modifications can be made to the embodiments of the present invention without departing from the principles described herein, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound represented by Formula (I), stereoisomers, and pharmaceutically acceptable salts thereof, 2. A pharmaceutical composition comprising the compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, and one or more pharmaceutically acceptable carriers or excipients.

3. The pharmaceutical composition of claim 2, wherein, The pharmaceutical composition further comprises one or more other therapeutic agents.

4. Use of the compound, pharmaceutically acceptable salt, stereoisomer according to claim 1 in the manufacture of a medicament for treating a cancer and an autoimmune disease associated with tyrosine kinase EGFR, wherein the cancer and the autoimmune disease comprise: ocular fundus diseases, dry eye, psoriasis, vitiligo, dermatitis, alopecia areata, rheumatoid arthritis, colitis, multiple sclerosis, systemic lupus erythematosus, Crohn's disease, atherosclerosis, pulmonary fibrosis, liver fibrosis, myelofibrosis, non-small cell lung cancer, small cell lung cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, ovarian cancer, cervical cancer, colorectal cancer, melanoma, endometrial cancer, prostate cancer, bladder cancer, leukemia, gastric cancer, liver cancer, gastrointestinal stromal tumor, thyroid cancer, chronic myelogenous leukemia, acute myelogenous leukemia, non-Hodgkin's lymphoma, nasopharyngeal carcinoma, esophageal cancer, brain tumor, B-cell and T-cell lymphomas, lymphoma, multiple myeloma, biliary carcinosarcoma, cholangiocarcinoma.

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