TWI820414B - Quinazoline compounds, preparation method and use thereof - Google Patents

Abstract

The present invention provides a quinazoline compound, preparation and use thereof, and specifically relates to a compound of formula (I), or isomers, hydrates, solvates, pharmaceutically acceptable salts and prodrugs thereof, and the preparation method thereof and the use thereof in the manufacture of a drug as tyrosine kinase inhibitors. The compound of the present invention has good inhibitory activity on EGFR and HER2 kinase, and at the same time exhibits excellent performance of penetrating the blood-brain barrier.

Description

Quinazoline compounds, preparation methods and applications

The invention belongs to the field of medical technology and relates to a quinazoline compound, a preparation method and its application.

Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein that belongs to the tyrosine kinase receptor family. It is widely expressed and plays an important role in growth and development and normal physiological functions. In addition, EGFR and the signaling pathways it mediates play an important role in the occurrence and development of tumors. However, the expression of EGFR is very unstable, and gene amplification and rearrangement often occur, which changes the antigen phenotype on the surface of tumor cells. Among them, epidermal growth factor receptor variant III (EGFRvIII) Most common.

EGFRvIII is a type of epidermal growth factor receptor (EGFR) mutant discovered in recent years that is only expressed on the surface of tumor cells but not on normal tissue cells. Abnormal expression of EGFR is associated with the occurrence of many malignant tumors, including glioma, lung small cell carcinoma, breast cancer, bladder cancer, ovarian cancer, etc.

Compared with the complete structure of EGFR, exons 2-7 encoding the extracellular ligand binding region of EGFRvIII are deleted, resulting in the deletion of 801 base pairs, allowing exons 1 and 8 to be connected, and binding at this A new glycine is generated at the site, resulting in the deletion of amino acids 6 to 273, thus losing the ability to bind to the ligand EGF. In the absence of ligand binding, EGFRvIII causes unregulated structural activation of tyrosine kinases through dimerization and autophosphorylation, inducing downstream signaling and stimulating tumor cell proliferation.

Existing studies have shown that EGFRvIII can affect the occurrence and development of tumors by regulating multiple signaling pathways, including Ras/Raf/MEK/ERK, PI3/AKT/mTOR, JAK/STAT and PLC/PKC, etc. The tumorigenicity of EGFRvIII-positive tumor cells is significantly increased, mainly by inhibiting cell apoptosis, promoting tumor angiogenesis, increasing invasiveness and migration, etc., leading to uncontrollable spontaneous proliferation and metastasis of tumor cells. In addition, EGFRvIII plays an escape-like function during tumor radiotherapy and chemotherapy.

Glioma is a common malignant tumor with high invasiveness, and glioblastoma (GBM) is the most malignant type. The effects of radiotherapy and chemotherapy are not ideal, and recurrences often occur after surgery. Domestic and foreign studies have found that 40% to 60% of GBM significantly express EGFR, and the mutant form is mainly EGFRvIII. EGFRvIII establishes a signaling pathway regulatory network through receptor-independent autophosphorylation and tyrosine kinase activity, and plays an important role in regulating the growth, metastasis and angiogenesis of GBM.

In recent years, studies have found that EGFRvIII molecularly targeted treatments have shown good anti-tumor effects in both in vitro cell culture and in vivo animal models. Therefore, the development of new molecularly targeted therapeutic drugs targeting EGFRvIII will provide more effective and economical treatment options for tumor patients, especially glioma patients, and there is a huge unmet clinical need.

Drugs targeting EGFRvIII to treat glioma not only need to be able to effectively penetrate the blood-brain barrier, but also need to be able to effectively inhibit EGFRvIII. There are currently no reports of compounds that can both penetrate the blood-brain barrier and inhibit EGFRvIII. Therefore, research on EGFRvIII-driven gliomas has important clinical value. In addition, the vast majority of marketed EGFR and HER2 kinase inhibitors are unable to penetrate the blood-brain barrier, and patients with EGFR-driven lung cancer and HER2-driven breast cancer generally have poor prognosis and a higher risk of brain metastasis. There are currently no effective drugs approved for the treatment of brain metastases, so there is an urgent need to develop an EGFR inhibitor and/or HER2 inhibitor that can penetrate the blood-brain barrier.

In one aspect, the present invention provides a compound represented by formula (I), its isomers, hydrates, solvates, pharmaceutically acceptable salts and prodrugs thereof, Formula (I) In formula (I), m is 0, 1 or 2; A is halogen, C ₁ -C ₃ alkyl; Z is NH or O; R ₁ is hydrogen, hydroxyl, 4-7 membered heteroalicyclic ring group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkyl substituted by hydroxyl, C C ₁ -C ₆ alkyl substituted by ₁ -C ₃ alkoxy, or C ₁ -C ₆ alkyl substituted by C ₃ -C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group contains 1-2 heteroalicyclic groups selected from N, O or S heteroatoms, the heteroalicyclic groups are unsubstituted or substituted by C ₁ -C ₃ alkyl, C ₁ -C ₄ alkyl acyl group, hydroxyl group _{In _ _ _ _ _} One or two substitutions; R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen, halogen, C ₁ -C ₆ alkyl, -O-(CH ₂ )nR ₇ , and R ₇ is Hydrogen, C ₁ -C ₃ alkyl, 1 to 3 selected from halogen, cyano group, hydroxyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, halogenated C ₁ -C ₃ alkyl _{n _ _ _ _ _} is an integer of 0-3, the aryl group is a monocyclic or bicyclic group containing 6 to 12 carbon ring atoms and at least one aromatic ring, and the heteroaryl group is a group containing 1-3 selected from N, O, S The heteroatoms in are used as ring atoms and are monocyclic or bicyclic groups containing 5 to 10 ring atoms, and R' and R'' are each independently H or a C ₁ -C ₃ alkyl group.

Or, when m is 1, the compound of formula (I) has the following general formula (II), Formula (II) In formula (II), A is halogen, C ₁ -C ₃ alkyl; Z is NH or O; R ₁ is hydrogen, hydroxyl, 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkyl substituted with hydroxyl, or C _{1 -} C ₃ alkoxy. C ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl substituted by C _{3 -} C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group contains 1-2 selected from N, Heteroalicyclic group of heteroatom of O or S, which is unsubstituted or substituted by C ₁ -C ₃ alkyl group, C ₁ -C ₄ alkyl group, amino group, hydroxyl group, cyano group, _{R _ _ _ _ _ 2.} R ₃ , R ₄ , R ₅ and R ₆ are each independently hydrogen, halogen, C ₁ -C ₆ alkyl, -O-(CH ₂ )nR ₇ , R ₇ is hydrogen, C ₁ -C ₃ alkyl Group, consisting of 1 to 3 selected from halogen, cyano group, hydroxyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, halogenated C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl aryl or heteroaryl substituted or unsubstituted by the substituents in the base, C ₂ -C ₃ alkynyl, C ₂ -C ₃ alkenyl or -NR'R'', n is an integer from 0 to 3, so The aryl group is a monocyclic or bicyclic group containing 6 to 12 carbon ring atoms and having at least one aromatic ring, and the heteroaryl group contains 1-3 heteroatoms selected from N, O, and S as ring atoms and A monocyclic or bicyclic group containing 5 to 10 ring atoms, R' and R'' are each independently H or a C ₁ -C ₃ alkyl group.

According to a preferred embodiment, A is Cl, F or methyl; Z is NH.

More preferably, A is Cl; Z is NH.

According to a preferred embodiment, R ₁ is a 4-7 membered heterocyclyl group or -NR ^a R ^b , and R ^a and R ^b are each independently hydrogen, C ₁ -C ₃ alkyl, or C ₁ - substituted by hydroxyl. C ₃ alkyl, C ₁ -C ₃ alkyl substituted by C ₁ -C ₃ alkoxy; the 4-7 membered heteroalicyclic group is pyrrolidinyl, pyridinyl, pyridinyl, morpholinyl , tetrahydrofuryl, tetrahydropyranyl, thiomorpholinyl, and the above groups are unsubstituted or replaced by methyl, ethyl, propyl, isopropyl, aldehyde, acetyl, propyl, hydroxyl , cyano, aminocarboxyl, methylprenyl, ethylprenyl, propylprenyl, isopropylprenyl, methylprenyl, ethylprenylene, propylprenyl, isopropyl One or two substitutions of stylene and oxo (=O).

More preferably, R ₁ is pyrrolidin-1-yl, pyridin-1-yl, 1-methylpyridin-4-yl, 1-ethylpyridin-4-yl, morpholinyl, tetrahydrofuran 2-yl base, tetrahydrofuran 3-yl, tetrahydropyran 2-yl, tetrahydropyran 3-yl, thiomorpholinyl, dimethylamino, diethylamino, dipropylamino, diisopropylamino, methylethyl Amino, methylpropylamino or ethylpropylamino.

Most preferably, _R1 is dimethylamino.

According to a preferred embodiment, m is 0 or 1, R ₁ is a 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₃ alkyl, C ₃ -C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group is pyrrolidinyl, piridyl, pyrazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl , and the above groups are unsubstituted or substituted by methyl, ethyl, propyl, isopropyl, aldehyde, acetyl, propyl, hydroxyl, cyano, aminocarboxylic, methylphenol, ethyl One or both of styrene, propylstyrene, isopropylstylene, methylstyrene, ethylstylene, propylstylene, isopropylstylene, oxo (=O) kind of replacement.

More preferably, m is 0 or 1, and R ₁ is 1-methyl-pyrrolidin-2-yl, 1-ethyl-pyrrolidin-2-yl, 1-isopropyl-pyrrolidin-2-yl, Methylamino, ethylamino, propylamino, isopropylamino, cyclopropylamino, cyclobutylamino, methylisopropylamino, N-methyl-N-cyclopropylamino, N-methyl -N-cyclobutylamino, pyrrolidin-1-yl, pyridin-1-yl, 1-methylpyridin-4-yl, 1-ethylpyridin-4-yl, morpholinyl, tetrahydrofuran 2 -yl, tetrahydrofuran 3-yl, tetrahydropyran 2-yl, tetrahydropyran 3-yl, thiomorpholinyl, dimethylamino, diethylamino, dipropylamino, diisopropylamino, methylethyl amino, methylpropylamino or ethylpropylamino.

According to another preferred embodiment, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ are each independently hydrogen, fluorine, chlorine, methyl, ethyl, propyl, isopropyl, -O-(CH ₂ ) nR ₇ , and at least 3 of R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are hydrogen, R ₇ is hydrogen, methyl, ethyl, propyl, isopropyl or 1 to 3 Selected from fluorine, chlorine, cyano, hydroxy, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl, trifluoromethyl, cyclopropyl, ethynyl, vinyl or -NR The substituent in 'R'' is a substituted or unsubstituted aryl or heteroaryl group, n is an integer from 0 to 3, R' and R'' are independently H or methyl, and the aryl group is benzene base, the heteroaryl group is pyridyl, pyrimidinyl, pyrrolyl, thienyl, furyl, imidazolyl.

More preferably, R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen, fluorine, chlorine, -O-(CH ₂ )nR ₇ , and R ₂ , R ₃ , R ₄ , R ₅ , at least 3 of R ₆ are hydrogen, R ₇ is 1 to 3 selected from fluorine, chlorine, cyano, hydroxyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl , an aryl group or a heteroaryl group substituted or unsubstituted by a substituent in the trifluoromethyl group, n is an integer from 0 to 3, the aryl group is a phenyl group, and the heteroaryl group is a pyridyl group.

In other preferred embodiments, R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen, fluorine, chlorine, phenoxy, 2-fluorophenoxy, 3-fluorophenoxy. , 4-fluorophenoxy, pyridin-2-ylmethoxy, pyridin-3-ylmethoxy, pyridin-4-ylmethoxy, 3-fluorobenzyloxy, 2-fluorobenzyloxy , 4-fluorobenzyloxy, 3-chlorobenzyloxy, 2-chlorobenzyloxy, 4-chlorobenzyloxy, and at least one of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ 3 are hydrogen.

According to another preferred embodiment, R ₂ , R ₃ , R ₅ , and R ₆ are each independently hydrogen, fluorine, chlorine, methyl, ethyl, propyl, isopropyl, and R ₄ is hydrogen, fluorine, chlorine , methyl, ethyl, propyl, isopropyl, -O-(CH ₂ )nR ₇ , and at least 2 of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are hydrogen, and R ₇ is hydrogen , methyl, ethyl, propyl, isopropyl or 1 to 3 selected from fluorine, chlorine, cyano, hydroxyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl A substituted or unsubstituted aryl or heteroaryl group, n is an integer from 0 to 3, R' , R'' are each independently H or methyl, the aryl group is phenyl, the heteroaryl group is pyridyl, pyrimidinyl, pyrrolyl, thienyl, furyl, imidazolyl; or, more preferably , R ₂ , R ₃ , R ₅ , and R ₆ are each independently hydrogen, fluorine, and chlorine, and R ₄ is hydrogen, fluorine, chlorine, phenoxy, 2-fluorophenoxy, 3-fluorophenoxy, 4 -Fluorophenoxy, pyridin-2-ylmethoxy, pyridin-3-ylmethoxy, pyridin-4-ylmethoxy, 3-fluorobenzyloxy, 2-fluorobenzyloxy, 4 -Fluorobenzyloxy, 3-chlorobenzyloxy, 2-chlorobenzyloxy, 4-chlorobenzyloxy, and at least 2 of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ is hydrogen.

Typical compounds involved in this application are as follows:

Another aspect of the invention provides a pharmaceutical composition comprising a compound described herein, a pharmaceutically acceptable salt, isomer, solvate, or prodrug thereof, and one or more pharmaceutically acceptable salts, isomers, solvates, or prodrugs thereof. Acceptable carrier or excipient.

Pharmaceutical compositions of the present application may also contain one or more other therapeutic agents.

The present invention also relates to a method for treating EGFR, HER2 and other kinase-mediated diseases or conditions, which includes administering a therapeutically effective amount of a compound described in the present application to a patient (human or other mammal, especially human) in need. Or a salt thereof. The diseases or conditions mediated by kinases such as EGFR and HER2 include those mentioned above.

Unless otherwise stated, the following terms used in this application (including the specification and claims) have the definitions given below. In this application, the use of "or" or "and" means "and/or" unless stated otherwise. Furthermore, the use of the term "includes" and other forms such as "includes," "contains," and "has" is not limiting. The section headings used in this article are for organizational purposes only and should not be construed as limitations on the subject matter described.

Unless otherwise specified, alkyl refers to a saturated straight-chain or branched hydrocarbon group with a specified number of carbon atoms. The term C ₁ -C ₁₀ alkyl refers to an alkyl moiety containing 1 to 10 carbon atoms. Similarly, C ₁ -C ₃ Alkyl represents an alkyl moiety containing 1 to 3 carbon atoms, for example, C ₁ -C ₆ 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-methyl Pentyl etc.

When a substituent term such as "alkyl" is used in combination with other substituent terms, for example in the term "C ₁ -C ₃ alkoxy C ₁ -C ₆ alkylthio" or "hydroxy substituted C ₁ -C ₁₀ alkyl ", the linking substituent term (eg, alkyl or alkylthio) is intended to include the divalent moiety wherein the point of attachment is through the linking substituent. Examples of "C ₁ -C ₃ alkoxy C ₁ -C ₆ alkylthio" include, but are not limited to, methoxymethylthio, methoxyethylthio, ethoxypropylthio, and the like. Examples of "hydroxy-substituted C ₁ -C ₁₀ alkyl" include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxyisopropyl, and the like.

Alkoxy is an alkyl-O- group formed from a previously described linear or branched alkyl group and -O-, for example, methoxy, ethoxy, and the like. Similarly, an alkylthio group is an alkyl-S- group formed from a linear or branched chain alkyl group and -S- as previously described, for example, methylthio, ethylthio, and the like.

Alkenyl and alkynyl include straight-chain, branched-chain alkenyl or alkynyl, and the term C ₂ -C ₆ alkenyl or C ₂ -C ₆ alkynyl represents a straight-chain or branched hydrocarbon group having at least one alkenyl or alkynyl group.

The term "haloalkyl", for example "halo C ₁ -C ₁₀ alkyl" means having one or more halogens, which may be the same or different, on one or more carbon atoms of the alkyl moiety including 1 to 10 carbon atoms. groups of atoms. Examples of "halo C ₁ -C ₁₀ alkyl" may include, but are not limited to -CF ₃ (trifluoromethyl), -CCl ₃ (trichloromethyl), 1,1-difluoroethyl, 2,2 , 2-trifluoroethyl and hexafluoroisopropyl, etc. Similarly, the term "halo C ₁ -C ₁₀ alkoxy" means a haloalkyl-O- group formed from said halo C ₁ -C ₁₀ alkyl and -O-, which may be, for example, trifluoromethyl Oxygen, trichloromethoxy, etc.

The term "C ₁ -C ₃ acyl" includes formyl (-CHO), acetyl (CH ₃ CO-), acetyl (C ₂ H ₅ CO-).

"Cycloalkyl" means a nonaromatic, saturated, cyclic hydrocarbon group containing the specified number of carbon atoms. For example, the term "(C ₃ -C ₆ )cycloalkyl" refers to a nonaromatic, cyclic hydrocarbon ring having 3 to 6 ring carbon atoms. Exemplary "(C ₃ -C ₆ ) cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term "aryl" means a group or moiety containing aromatic monocyclic or bicyclic hydrocarbon radicals containing 6 to 12 carbon ring atoms and having at least one aromatic ring. Examples of "aryl" are phenyl, naphthyl, indenyl and indenyl (indanyl). Typically, in the compounds of the invention, the aryl group is phenyl.

The term "heteroalicyclic" as used herein, unless otherwise specified, represents an unsubstituted or substituted stable 4 to 8 membered nonaromatic monocyclic saturated ring system consisting of carbon atoms and from N, It consists of 1 to 3 heteroatoms selected from O and S. Among them, N and S heteroatoms can be oxidized at will, and N heteroatoms can also be quaternized at will. Examples of such heterocycles include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, pyrazolidinyl, pyrazolinyl, imidazole Alkyl, imidazolinyl, oxazolinyl, thiazolinyl, tetrahydrofuryl, dihydrofuryl, tetrahydrothienyl, 1,3-dioxolyl, piridinyl, pyridinyl, tetrahydrofuranyl Hydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, 1,3-dioxanyl, 1,4-dioxanyl, 1,3-oxothiolanyl, 1,3 -Oxathiolyl, 1,3-dithianyl, 1,4-oxathiolyl, 1,4-oxothianyl, 1,4-dithianyl , morpholinyl, thiomorpholinyl.

As used herein, the term "heteroaryl" means a group or moiety containing an aromatic monocyclic or bicyclic group of atoms (containing 5 to 10 ring atoms) containing 1 to 3 atoms independently selected from nitrogen, oxygen, and sulfur. of heteroatoms. The term also includes bicyclic heterocyclic aryl groups containing an aryl ring moiety fused to a heterocycloalkyl ring moiety or a heteroaryl ring moiety fused to a cycloalkyl ring moiety. Unless otherwise specified, it represents an unsubstituted or substituted stable 5- or 6-membered monocyclic aromatic ring system, or an unsubstituted or substituted benzene-condensed heteroaromatic ring system with 9 or 10 ring atoms. Or bicyclic heteroaromatic ring systems, which are composed of carbon atoms and 1 to 3 heteroatoms selected from N, O, and S. The N and S heteroatoms can be oxidized, and the N heteroatoms can also be quaternized. . Heteroaryl groups can be linked to any heteroatom or carbon atom to form a stable structure. Illustrative examples of heteroaryl groups include, but are not limited to, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazole base, thiadiazolyl, isothiazolyl, pyridyl, oxo-pyridyl (pyridyl-N-oxide), pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, benzofuryl, iso Benzofuranyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolyl, dihydrobenzodioxenyl, benzothienyl, indolizinyl , indolyl, isoindolyl, indolyl, benzimidazolyl, dihydrobenzimidazolyl, benzoxazolyl, dihydrobenzoxazolyl, benzothiazolyl, benziiso Thiazolyl, dihydrobenzisothiazolyl, indazolyl, imidazopyridyl, pyrazolopyridyl, benzotriazolyl, triazolopyridyl, purinyl, quinolyl, tetrahydroquinolinyl , isoquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl, 1,5-diazanaphthyl, 1,6-diazanaphthyl , 1,7-diazanaphthyl, 1,8-diazanaphthyl and pteridinyl.

The term "carbonyl" refers to a -C(O)- group. The terms "halogen" and "halo" refer to chlorine, fluorine, bromine or iodine substituents. "Oxo" refers to the oxygen portion of a double bond; for example, if attached directly to a carbon atom to form a carbonyl moiety (C=O). "Hydroxy" is intended to mean the -OH radical. As used herein, the term "cyano" refers to the group -CN.

The term "each independently" means that when more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.

It will be understood that the compounds, isomers, crystal forms or prodrugs of formula I and pharmaceutically acceptable salts thereof may exist in solvated and unsolvated forms. For example, the solvated form may be a water-soluble form. The invention includes all such solvated and unsolvated forms.

The term "isomer" in this application refers to different compounds with the same molecular formula, which may include various isomeric forms such as stereoisomerism and tautomerism. "Stereoisomers" are isomers that differ only in the arrangement of their atoms in space. Certain compounds described herein contain one or more asymmetric centers and, therefore, may give rise to enantiomers, diastereomers, and other stereoisomers that may be defined as (R)- or (S)- based on absolute stereochemistry. form. The chemical entities, pharmaceutical compositions and methods of the invention are intended to encompass all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optical (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of a compound can be analyzed by any suitable method, including, but not limited to, chiral chromatography and polarimetry, and the degree of dominance of one stereoisomer over other isomers can be determined.

Individual isomers (or isomer-enriched mixtures) of the present invention can be resolved using methods known to those skilled in the art. For example, the resolution can be performed: (1) by the formation of diastereomeric salts, complexes or other derivatives; (2) by selective reaction with stereoisomer-specific reagents, such as by enzymes Promotes oxidation or reduction; or (3) by gas-liquid chromatography or liquid chromatography in a chiral environment, such as on a chiral support (such as silica bound with chiral ligands) or on a hand in the presence of sexual solvents. Those skilled in the art will understand that when the desired stereoisomer is converted into another chemical entity by one of the separation methods described above, additional steps are required to release the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by asymmetric transformation to convert one enantiomer to the other.

When a compound described herein contains an olefinic double bond, it is intended that the compound includes various cis and trans isomers, unless otherwise stated.

"Tautomers" are structurally distinct isomers that are interconvertible by tautomerization. "Tautomerization" is a form of isomerization and includes proton transfer or proton transfer tautomerization, which can be considered a subset of acid-base chemistry. "Proton transfer tautomerization" or "proton transfer tautomerization" involves proton migration accompanied by bond order transformation, often the exchange of a single bond with an adjacent double bond. When tautomerization is possible (eg, in solution), a chemical equilibrium of the tautomers can be achieved. An example of tautomerization is keto-enol tautomerization.

The compounds of the invention as active ingredients, as well as methods for preparing the compounds, are within the scope of the present invention. Furthermore, crystalline forms of some compounds may exist as polycrystals, and such forms may also be included in the present invention. In addition, some compounds can form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are also included in the scope of this invention.

The compounds of the invention may be used in therapy in free form or, where appropriate, in the form of pharmaceutically acceptable salts or other derivatives. As used herein, the term "pharmaceutically acceptable salts" refers to organic salts and inorganic salts of the compounds of the present invention, which salts are suitable for humans and lower animals, have no excessive toxicity, irritation, allergic reactions, etc., and have reasonable Benefit/risk ratio. Pharmaceutically acceptable salts of amines, carboxylic acids, phosphonates, and other types of compounds are well known in the art. The salts may be formed by reacting a compound of the invention with a suitable free alkaloid or acid. Including, but 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, Alternatively, these salts can be obtained using methods well known in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate , camphorsulfonate, citrate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate , hemisulfate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, methanesulfonic acid Salt, 2-naphthalene sulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectate, persulfate, per3-phenylpropionate, phosphate , picrate, propionate, stearate, sulfate, thiocyanate, p-toluenesulfonate, undecanoate, etc. Representative alkaline or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts include the appropriate nontoxic ammonium, quaternary ammonium, and salts using salts such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and arylsulfonic acids. Salt-forming amine cation.

In addition, the term "prodrug" as used herein refers to a compound that can be converted in vivo into a compound of the present invention. This conversion is affected by hydrolysis of the prodrug in the blood or enzymatic conversion to the parent compound in the blood or tissue.

The pharmaceutical composition of the present invention includes compounds described herein or pharmaceutically acceptable salts thereof, kinase inhibitors (small molecules, polypeptides, antibodies, etc.), immunosuppressants, anticancer drugs, antiviral agents, anti-inflammatory agents, anti- A fungative, antibiotic, or additional active agent of the anti-angiogenic compound; and any pharmaceutically acceptable carrier, adjuvant, or excipient.

The compounds of the present invention may 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 may be formulated for administration simultaneously or sequentially at different times, or the therapeutic agents may be administered as a single composition. The so-called "combination therapy" refers to the use of a compound of the present invention together with another agent. The administration method is that each agent is co-administered at the same time or each agent is administered sequentially. In either case, the purpose is to achieve Optimum effect of the drug. Co-administration includes simultaneous delivery of dosage forms, as well as separate separate dosage forms for each compound. Therefore, the administration of the compounds of the present invention may be used concurrently with other therapies known in the art, for example, in cancer treatment using radiation therapy or additional therapies such as cytostatic agents, cytotoxic agents, other anti-cancer agents, etc. to improve Cancer symptoms. The invention is not limited to the order of administration; the compounds of the invention may be administered previously, simultaneously, or after other anticancer or cytotoxic agents.

In order to prepare the pharmaceutical composition of this invention, one or more compounds or salts of the molecular formula (I) as its active ingredient can be closely mixed with a pharmaceutical carrier, which is carried out according to traditional pharmaceutical compounding technology, wherein The carrier can take a variety of forms depending on the form of preparation designed for different modes of administration (eg, oral or parenteral administration). Suitable pharmaceutically acceptable carriers are well known in the art. A description of some of these pharmaceutically acceptable carriers can be found in the Handbook of Pharmaceutical Excipients, published jointly by the American Pharmaceutical Association and the British Pharmaceutical Society.

The pharmaceutical composition 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 liquids, suspensions, emulsions Liquid; or for topical application such as ointments, creams; or as suppositories for rectal administration. Pharmaceutical ingredients may also be available in unit dose form suitable for single-use administration of precise doses. The pharmaceutical composition will include a traditional pharmaceutical carrier or excipient and a compound prepared according to the current invention as an active ingredient. In addition, it may also include other medical or pharmaceutical preparations, carriers, auxiliaries, etc.

The therapeutic compounds may also be administered to mammals other than humans. The dosage of the drug administered to a mammal will depend on the species of the animal and its disease or disorder. Therapeutic compounds can be fed to animals in the form of capsules, bolus pills, tablet solutions. Therapeutic compounds can also be introduced into animals by injection or infusion. We prepare these pharmaceutical forms according to traditional methods consistent with standards of veterinary practice. As an alternative, pharmaceutical compounds can be mixed with animal feed and fed to animals, so that concentrated feed additives or ready-mixes can be prepared for mixing with ordinary animal feed.

It is a further object of the present invention to provide a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition containing a compound of the present invention.

The present invention also includes the use of the compound of the present invention or its pharmaceutically acceptable derivatives in the preparation of drugs for the treatment of cancer and autoimmune diseases related to tyrosine kinase EGFR and HER2. Such cancers (including non-solid tumors, solid tumors, primary or metastatic cancers, as noted elsewhere herein and including cancers resistant or refractory to one or more other treatments) as well as other diseases (including but not limited to fundus diseases, psoriasis, atherosclerosis, pulmonary fibrosis, liver fibrosis, myelofibrosis, etc.). The cancers 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 myelogenous leukemia, acute myeloid leukemia, non-Hodgkin lymphoma, nasopharyngeal cancer, esophageal cancer, brain tumor, B Any of cell and T-cell lymphoma, lymphoma, multiple myeloma, biliary carcinosarcoma, cholangiocarcinoma.

Detailed implementation

The present invention also provides methods for preparing corresponding compounds. Various synthetic methods can be used to prepare the compounds described herein, including the following methods. The compounds of the present invention or their pharmaceutically acceptable salts, isomers or hydrates can be used The following methods are either synthetic methods known in the field of organic chemical synthesis, or are synthesized by variations of these methods understood by those skilled in the art. Preferred methods include but are not limited to the following methods.

In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention and are not intended to limit the present invention. If specific techniques or conditions are not specified in the examples, the techniques or conditions described in literature in the field or product instructions will be followed. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. The examples provided below can better illustrate the present invention. Unless otherwise specified, all temperatures are °C. The names of some of the compounds in this application are translated according to chemdraw nomenclature.

Synthesis of intermediates

Synthesis of acid intermediates: (E)-4-(dimethylamino)but-2-enoic acid, (E)-4-(diethylamino)but-2-enoic acid and (E)-4-( Synthetic method of pyridin-1-yl)but-2-enoic acid

Take (E)-4-(diethylamino)but-2-enoic acid as an example. Under argon protection, add 2 g (27.34 mm) diethylamine and 20 ml THF to the flask, add 5 g (27.93 mm) methyl bromate and 9 g (69.63 mm) diisopropyl under ice bath Ethylamine. React for 3 hours, extract with ethyl acetate and water; remove the solvent by rotary evaporation, add liquid salt (2 g NaOH and 2 ml H ₂ O) to 20 ml of ethanol, stir for 3 hours, add concentrated hydrochloric acid dropwise to adjust the pH to 1-2, concentrate under reduced pressure. Acetone (20 ml) was added and recrystallized to give a white solid.

Preparation of (R,E)-3-(1-methylpyrrolidin-2-yl)acrylyl chloride

Add (R,E)-3-(1-methylpyrrolidin-2-yl)acrylic acid (160mg, 1mmol) to dry dichloromethane (3ml), add oxalyl chloride (130mg, 1mmol) and DMF respectively. (1 drop, catalytic amount), stir at room temperature for 3 hours, the reaction system becomes turbid and becomes clear, concentrate to obtain an off-white solid;

Synthesis of amine intermediates B1-B5:

Synthesis of 4-((1H-benzo[d][1,2,3]triazol-1-yl)oxy)-5-chloroquinazolin-6-amine

Conditions and reagents: (a) EtOH, formamidine acetate, 80 ℃, 8h; (b) H ₂ SO ₄ , HNO ₃ , -5 ℃, overnight; (c) CH ₃ OH, Fe, NH ₄ Cl, 80 ℃ , reflux/Ni, H ₂ , rt; (d) CH ₃ CN, BOP, DBU, rt

Using 2-amino-6-chlorobenzoic acid as the starting material, cyclization was performed using formamidine acetate at 80°C to obtain the compound 5-chloroquinazolin-4(3H)-one, which was then dissolved in concentrated sulfuric acid. To the solution, nitric acid was added at -10°C, and the compound 5-chloro-6-nitroquinazolin-4(3H)-one was obtained by column chromatography. Then we use iron powder to reduce the nitration product, and obtain the compound 5-chloro-6-aminoquinazolin-4(3H)-one in an acidic environment (or use Raney nickel to reduce the nitration product in a hydrogen environment). Then, by using BOP, 4-((1H-benzo[d][1,2,3]triazol-1-yl)oxy)-5-chloroquinazolin-6-amine is obtained.

Compound 4-((1H-benzo[d][1,2,3]triazol-1-yl)oxy)-5-chloroquinazolin-6-amine (100 mg, 0.32 mM) and - A series of amine compounds (71-90 mg, 0.38 mM) were dissolved in i-PrOH (20 ml) solution and stirred at 90°C for 10 minutes, then TsOH (7 mg, 0.03mM) was added to the mixture and reacted for 5 hours. . And monitor the reaction progress by TLC, at the end of the reaction, add water and filter to obtain a solid. Intermediates B1-B5 (brown or green solids) were purified by column chromatography (EA:PE = 5:1). Their structures and characterizations are shown in Table 1 below.

Table 1 Structure, naming and characterization of intermediates B1-B5 Intermediate number structure name LCMS m/z = (M+H) ⁺ B1 5-Chloro-N4-(4-phenoxyphenyl)quinazoline-4,6-diamine 363.1 B2 5-Chloro-N4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)quinazoline-4,6-diamine 412.1 B3 5-Chloro-N4-(4-((3-fluorobenzyl)oxy)phenyl)quinazoline-4,6-diamine 395.1 B4 5-Chloro-N4-(3-chloro-2-fluorophenyl)quinazoline-4,6-diamine 323.0 B5 5-Chloro-N4-(3-chloro-4-fluorophenyl)quinazoline-4,6-diamine 323.0

Example

Synthesis method one:

Serial 2-enoic acid (120 mg, 0.9 mM) was dissolved in anhydrous DCM (5 ml), and oxalate chloride (80 μL, 0.9 mM) was added under ice bath for 3 h. The solvent was then evaporated to give an orange solid. Then the orange solid dissolved in DCM (1 ml) was added to a solution of B1-B5 (100 mg, 0.3 mM) in NMP (2 ml) under ice bath for 4 h. And monitor the reaction progress by TLC. Potassium carbonate solution was then added to adjust the pH to 8-9, and the crude product was extracted with DCM. Purify by column chromatography (DCM: _CH3OH =30:1-10:1). Examples 1-7 were synthesized using this method.

Synthesis method two:

Dissolve (E)-4-bromobut-2-enoic acid (150 mg, 0.9 mM) in anhydrous DCM (5 ml), and add oxalate chloride (80 μL, 0.9 mM) in an ice bath for 3 h. The solvent was then evaporated to give an orange solid. Then the orange solid dissolved in DCM (1 ml) was added to a solution of B1-B5 (100 mg, 0.3 mM) in NMP (2 ml) under ice bath for 4 h. And monitor the reaction progress by TLC. Potassium carbonate solution was then added to adjust the pH to 8-9, and the crude product was extracted with DCM. The pure product was isolated by column chromatography (EA:PE = 5:1-3:1). Then, pyrrolidine (100-150 mg, 1.5 mM) was added to the product in DMA solution and stirred at 0 °C for 4 h. And monitor the reaction progress by TLC. Potassium carbonate solution was then added to adjust the pH to 8-9, and the crude product was extracted with DCM. The pure product was isolated by column chromatography (DCM: CH3OH = 30:1-10:1). Examples 8-10 were synthesized using this method two.

Example 1. (E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl)-4 -(Dimethylamino)but-2-enamide

White solid; mp: 230-232 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 9.98 (s, 1H), 9.51 (s, 1H), 8.53 (d, J = 4.8 Hz, 1H), 8.45 (s, 1H), 8.05 (d, J = 9.0 Hz, 1H), 7.84 – 7.81 (m, 2H), 7.68 (d, J = 9.0 Hz, 1H), 7.52 (d, J = 7.8 Hz, 2H), 7.33 – 7.29 (m, 1H), 7.20 (d, J = 9.0 Hz, 1H), 6.75 (dt, J = 15.5, 5.9 Hz, 1H), 6.45 (d, J = 15.5 Hz, 1H ), 5.24 (s, 2H), 3.03 (d, J = 5.6 Hz, 2H), 2.13 (s, 6H). ¹³ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.35 (s), 157.23 (s), 156.74 (s), 154.22 (s), 150.81 (s), 149.61 (s), 143.11 (s), 137.63 (s), 134.45 (s), 132.84 (s), 131.97 (s), 130.12 (s), 127.46 (s), 125.59 (s), 125.50 (s), 123.90 (s), 123.59 (s), 121.96 (s), 121.52 (s), 114.67 (s), 113.36 (s), 71.66 (s), 60.20 (s), 45.62 (s). HRMS (ESI) m/z Calculated for C ₂₆ H ₂₄ Cl ₂ N ₆ O ₂ ⁺ [M+H] ⁺ , 523.1416; Found, 523.1456.

Example 2. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(dimethylamino)butan-2 -enamide

White solid; mp: 235-237 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.02 (s, 1H), 9.67 (s, 1H), 8.53 (s, 1H), 8.16 ( d, J = 8.9 Hz, 1H), 7.79 (s, 2H), 7.46 (s, 1H), 7.29 (t, J = 7.8 Hz, 1H), 6.83 (dt, J = 15.4, 5.8 Hz, 1H), 6.53 (d, J = 15.5 Hz, 1H), 3.10 (dd, J = 5.8, 1.1 Hz, 2H), 2.20 (s, 6H). ¹³ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.30 (s), 153.49 (s), 153.07 (s), 143.15 (s), 134.72 (s), 132.00 (s), 130.10 (s), 127.10 (s), 126.08 (s), 125.49 (s), 125.45 (s), 125.40 (s), 121.47 (s), 120.39 (s), 120.22 (s), 113.69 (s), 109.82 (s), 60.24 (s), 45.66 (s). HRMS (ESI) m /z Calculated for C ₂₀ H ₁₈ Cl ₂ FN ₅ O ⁺ [M+H] ⁺ , 434.0951; Found, 434.0923.

Example 3. (E)-N-(5-chloro-4-((3-chloro-4-fluorophenyl)amino)quinazolin-6-yl)-4-(dimethylamino)butan-2 -enamide

White solid; mp: 221-223 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.01 (s, 1H), 9.66 (s, 1H), 8.59 (d, J = 16.7 Hz, 1H), 8.16 (d, J = 9.0 Hz, 1H), 8.04 (d, J = 4.3 Hz, 1H), 7.79 – 7.66 (m, 2H), 7.45 (t, J = 9.0 Hz, 1H), 6.82 ( dt, J = 15.4, 5.8 Hz, 1H), 6.54 (d, J = 15.5 Hz, 1H), 3.10 (d, J = 5.4 Hz, 2H), 2.20 (s, 6H). ¹³ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.43 (s), 156.77 (s), 150.82 (s), 149.61 (s), 144.08 (s), 137.60 (s), 134.56 (s), 132.03 (s), 130.11 (s), 127.45 (s), 125.56 (s), 125.11 (s), 123.87 (s), 123.57 (s), 121.94 (s), 121.55 (s), 114.71 (s), 71.72 (s), 47.17 (s). HRMS (ESI) m/z calcd for C ₂₀ H ₁₈ Cl ₂ FN ₅ O ⁺ [M+H] ⁺ , 434.0951; found, 434.0984.

Example 4. (E)-N-(5-chloro-4-((4-phenoxyphenyl)amino)quinazolin-6-yl)-4-(diethylamino)but-2-ene amide

White solid; mp: 236-238 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.31 (d, J = 8.3 Hz, 1H), 9.64 (s, 1H), 8.51 (s, 1H), 8.08 (d, J = 9.0 Hz, 1H), 7.77 – 7.71 (m, 3H), 7.40 (t, J = 7.9 Hz, 2H), 7.12 (d, J = 7.4 Hz, 1H), 7.07 ( d, J = 8.9 Hz, 2H), 7.04 – 6.99 (m, 2H), 6.92 – 6.85 (m, 1H), 6.66 (d, J = 14.9 Hz, 1H), 3.29 (d, J = 7.1 Hz, 2H ), 2.69 (s, 4H), 1.08 (t, J = 7.0 Hz, 6H), ¹³ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.11 (s), 157.58 (s), 157.21 ( s), 154.29 (s), 153.42 (s), 150.16 (s), 134.56 (s), 134.48 (s), 132.17 (s), 130.51 (s), 130.10 (s), 129.11 (s), 127.42 ( HRMS ( ESI) m/z calculated for C ₂₈ H ₂₈ ClN ₅ O ₂ ⁺ [M+H] ⁺ , 502.2010; found, 502.2009.

Example 5. (E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl)-4 -(Piridin-1-yl)but-2-enamide

White solid; mp: 240-242 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.03 (s, 1H), 9.57 (s, 1H), 8.55 (d, J = 36.7 Hz, 2H), 8.11 (d, J = 9.0 Hz, 1H), 7.95 – 7.83 (m, 2H), 7.73 (d, J = 8.9 Hz, 1H), 7.58 (d, J = 8.0 Hz, 2H), 7.42 – 7.32 (m, 1H), 7.26 (d, J = 9.0 Hz, 1H), 6.82 (dt, J = 15.3, 5.8 Hz, 1H), 6.52 (d, J = 15.9 Hz, 1H), 5.30 (s, 2H ), 3.10 (d, J = 5.6 Hz, 2H), 2.36 (s, 4H), 1.59 – 1.34 (m, 6H). ¹³ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.35 (s ), 157.18 (s), 156.75 (s), 154.17 (s), 150.81 (s), 149.57 (s), 143.00 (s), 137.58 (s), 134.50 (s), 132.86 (s), 131.94 (s) ), 127.48 (s), 125.56 (s), 125.52 (s), 123.81 (s), 123.57 (s), 123.25 (s), 121.94 (s), 121.56 (s), 121.45 (s), 114.66 (s) ), 71.71 (s), 59.72 (s), 54.56 (s), 25.94 (s), 24.27 (s). HRMS (ESI) m/z Calculated for C ₂₉ H ₂₈ Cl ₂ FN ₆ O ₂ ⁺ [M+ H] ⁺ , 563.1729; measured value, 563.1796.

Example 6. (E)-N-(5-chloro-4-((4-((3-fluorobenzyl)oxy)phenyl)amino)quinazolin-6-yl)-4-(gua (Din-1-yl)but-2-enamide

White solid; mp: 197-199 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.06 (s, 1H), 9.57 (s, 1H), 8.46 (s, 1H), 8.08 ( d, J = 9.0 Hz, 1H), 7.72 (d, J = 9.0 Hz, 1H), 7.60 (d, J = 8.9 Hz, 2H), 7.45 (td, J = 8.0, 6.1 Hz, 1H), 7.30 ( t, J = 7.8 Hz, 2H), 7.16 (td, J = 8.6, 2.2 Hz, 1H), 7.06 (d, J = 9.0 Hz, 2H), 6.82 (dt, J = 15.4, 5.9 Hz, 1H), 13 _ ^_ C NMR (101 MHz, DMSO-d ₆ , δ ppm): δ 164.45 (s), 163.48 (s), 161.86 (s), 155.61 (s), 154.35 (s), 149.96 (s), 143.11 (s) , 140.57 (s), 140.52 (s), 134.27 (s), 131.91 (s), 130.96 (d, J = 8.3 Hz), 127.36 (s), 125.48 (s), 123.86 (d, J = 2.5 Hz) , 115.38 (s), 115.05 (s), 114.91 (s), 114.59 (s), 114.45 (s), 113.26 (s), 69.01 (s), 59.72 (s), 54.52 (s), 25.86 (s) , 24.22 (s). HRMS (ESI) m/z Calculated for C ₃₀ H ₂₉ ClFN ₅ O ₂ ⁺ [M+H] ⁺ , 546.2072; Found, 546.2019.

Example 7. (E)-N-(5-chloro-4-((3-chloro-4-fluorophenyl)amino)quinazolin-6-yl)-4-(pyridin-1-yl) But-2-enamide

White solid; mp: 233-235 ℃. ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.05 (s, 1H), 9.68 (s, 1H), 8.53 (s, 1H), 8.12 ( d, J = 9.0 Hz, 1H), 8.01 (s, 1H), 7.80 – 7.61 (m, 2H), 7.49 – 7.39 (m, 1H), 6.82 (dt, J = 15.4, 5.9 Hz, 1H), 6.52 (d, J = 15.7 Hz, 1H), 3.10 (d, J = 5.5 Hz, 2H), 2.35 (s, 4H), 1.45 (d, J = 52.9 Hz, 6H). ¹³ C NMR (101 MHz, DMSO -d ₆ , δ ppm): δ 164.31 (s), 157.07 (s), 154.02 (s), 150.11 (s), 143.17 (s), 136.19 (s), 134.70 (s), 132.07 (s), 127.54 (s), 125.35 (d, J = 17.8 Hz), 124.22 (s), 121.24 (s), 119.35 (s), 117.20 (s), 116.98 (s), 113.43 (s), 59.76 (s), 54.62 (s), 26.02 (s), 24.32 (s). HRMS (ESI) m/z Calculated for C ₂₃ H ₂₂ Cl ₂ FN ₅ O ⁺ [M+H] ⁺ , 474.1264; Found, 474.1271.

Example 8. (E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl)-4 -(pyrrolidin-1-yl)but-2-enamide

White solid; ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.04 (s, 1H), 9.57 (s, 1H), 8.61 (d, J = 3.3 Hz, 1H), 8.53 (s, 1H), 8.12 (d, J = 9.0 Hz, 1H), 7.96 – 7.85 (m, 2H), 7.76 (d, J = 9.0 Hz, 1H), 7.59 (d, J = 6.8 Hz, 2H), 7.38 ( dd, J = 6.8, 5.1 Hz, 1H), 7.28 (d, J = 9.0 Hz, 1H), 6.86 (dt, J = 15.3, 5.7 Hz, 1H), 6.56 (d, J = 15.6 Hz, 1H), 5.32 (s, 2H), 3.33 – 3.28 (m, 2H), 2.66 (dd, J = 15.3, 13.6 Hz, 4H), 1.76 (s, 4H). MS: 548.9 [M+H] ⁺ .

Example 9. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(pyrrolidin-1-yl) But-2-enamide

White solid; ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.10 (s, 1H), 9.65 (s, 1H), 8.55 (s, 1H), 8.17 (d, J = 9.0 Hz, 1H), 7.83 (dd, J = 13.2, 8.7 Hz, 1H), 7.57 – 7.43 (m, 1H), 7.37 – 7.24 (m, 1H), 6.86 (dd, J = 13.6, 7.7 Hz, 1H), 5.76 (s, 1H), 5.33 (d, J = 4.7 Hz, 1H), 3.31 (s, 2H), 2.68 (d, J = 1.8 Hz, 4H), 1.24 (s, 4H). HRMS (ESI) m/ z Calculated for C ₂₂ H ₂₀ Cl ₂ FN ₅ O ⁺ [M+H] ⁺ , 460.1107; Found, 460.1101.

Example 10. (E)-N-(5-chloro-4-((3-chloro-4-fluorophenyl)amino)quinazolin-6-yl)-4-(pyrrolidin-1-yl) But-2-enamide

White solid; ¹ H NMR (400 MHz, DMSO-d ₆ , δ ppm): δ 10.04 (s, 1H), 9.67 (s, 1H), 8.58 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 8.05 (dd, J = 6.8, 2.6 Hz, 1H), 7.78 (d, J = 9.0 Hz, 1H), 7.71 (ddd, J = 8.9, 4.3, 2.7 Hz, 1H), 7.47 (t, J = 9.1 Hz, 1H), 6.87 (dt, J = 15.4, 5.7 Hz, 1H), 6.56 (d, J = 15.4 Hz, 1H), 3.32 – 3.28 (m, 2H), 2.70 – 2.58 (m, 4H) , 1.75 (s, 4H). HRMS (ESI) m/z calculated for C ₂₂ H ₂₀ Cl ₂ FN ₅ O ⁺ [M+H] ⁺ , 460.1107; found, 460.1100.

Example 11. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(isopropylamino)butan- 2-enamide

Step 1) Synthesis of 5-chloro-N-(3-chloro-2-fluorophenyl)-6-nitroquinazolin-4-amine

Suspend 5-chloro-6-nitroquinazolin-4-ol (1 g, 4.5 mmol) in triturous chloride (15 mL), add DMF (0.5 mL) under stirring at room temperature, and then heat the system to React at 105°C. After the system is clarified (about 3h), heat to 90°C and reflux for 2h. Monitor with LCMS (MeOH quenching system). After the reaction is completed, the system is directly concentrated under reduced pressure to obtain a light yellow solid; the above obtained The solid (1 g, 4.4 mmol) was suspended in dry acetonitrile (15 mL), sonicated to disperse evenly. Under ice bath conditions, 3-chloro-2-fluoroaniline (2.9 g, 20 mmol) was slowly added dropwise, and the dripping was completed. , remove the ice bath, heat to 50°C for 2 hours, monitor the reaction with LCMS to complete, concentrate, add MeOH to beat, filter, collect the filter cake to obtain the target product 850 mg, yield 53%, MS: 353[M+H] ⁺ ;

Step 2) Synthesis of 5-chloro-N-(3-chloro-2-fluorophenyl)quinazoline-4,6-diamine

5-Chloro-N-(3-chloro-2-fluorophenyl)-6-nitroquinazolin-4-amine (350mg, 1mmol), iron powder (280mg, 5mmol) and ammonium chloride (530mg, 10mmol) ) were added to a mixed solution of ethanol (10 ml) and water (1 ml), stirred and heated to 80 degrees Celsius for 1 hour, filtered through diatomaceous earth, the filtrate was washed with ethyl acetate and saturated sodium bicarbonate, the organic phase was dried and concentrated. 290 mg of off-white solid was obtained, which was directly used in the next step, MS: 323[M+H] ⁺ ;

Step 3) (E)-4-bromo-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)but-2-enamide synthesis

To a solution of 5-chloro-N-(3-chloro-2-fluorophenyl)quinazoline-4,6-diamine (65 mg, 0.2 mmol) in NMP (2 ml) under ice-water bath conditions, bromocroton was added Dichloromethane solution (55 mg, 0.3 mmol) was stirred for 30 minutes, then quenched by adding saturated sodium bicarbonate solution, and the solid precipitated, filtered, washed with ethyl acetate, dried and used directly in the next step; MS: 469 , 471[M+H] ⁺ ;

Step 4) (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(isopropylamino)butan-2 -Synthesis of enamides

(E)-4-bromo-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)but-2-ene under ice-water bath conditions Amide (0.46g, 1mmol), isopropylamine (0.2g, 3mmol) and diisopropylethylamine (0.3g, 3mmol) were added to DMF (5ml) respectively, heated to 50 degrees Celsius, stirred for 2 hours, and cooled. Water and ethyl acetate were added respectively, and the organic phase was washed with saturated brine, dried, concentrated, and purified by column chromatography to obtain 150 mg of light yellow solid product, with a yield of 33%;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.18 (s, 1H), 9.66 (s, 1H), 8.56 (s, 1H), 8.14 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 8.9 Hz, 2H), 7.49 (t, J = 7.6 Hz, 1H), 7.31 (t, J = 8.1 Hz, 1H), 6.89 (dt, J = 15.4, 5.8 Hz, 1H), 6.60 (d, J = 15.4 Hz, 1H), 3.63 (s, 2H), 3.33(br, 1H), 3.06 (s, 1H), 1.15 (d, J = 6.3 Hz, 6H). MS: 448 [M+H] ⁺ .

Example 12. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(cyclopropylamino)butan- 2-enamide

The synthesis is carried out in the same manner as in Example 11, except that cyclopropylamine is used instead of isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.00 (s, 1H), 9.65 (s, 1H), 8.54 (s, 1H), 8.16 (d, J = 9.0 Hz, 1H), 7.89 – 7.77 ( m, 2H), 7.49 (ddd, J = 8.3, 6.8, 1.6 Hz, 1H), 7.30 (td, J = 8.1, 1.4 Hz, 1H), 6.92 (dt, J = 15.4, 5.3 Hz, 1H), 6.49 (dt, J = 15.5, 1.8 Hz, 1H), 3.43 (dd, J = 5.4, 1.8 Hz, 2H), 3.33(br, 1H), 2.16 (tt, J = 6.7, 3.6 Hz, 1H), 0.43- .039 (m, 2H), 0.32 – 0.24 (m, 2H). MS: 446 [M+H] ⁺ .

Example 13. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(methylamino)butan-2- Enamide monotrifluoroacetate

The synthesis is carried out in the same manner as in Example 11, except that methylamine hydrochloride is used instead of isopropylamine in step 4) for reaction, and the monotrifluoroacetate product is prepared through preparative liquid phase purification;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.29 (s, 1H), 9.71 (s, 1H), 8.73 (s, 2H), 8.57(s, 1H), 8.12 (d, J = 9.1 Hz, 1H), 7.82 (s, 2H), 7.49 (s, 1H), 7.31 (d, J = 7.2 Hz, 1H), 6.82 (dt, J = 15.5, 6.5 Hz, 1H), 6.65 (d, J = 15.5 Hz, 1H), 3.84 (q, J = 5.9 Hz, 2H), 2.67 – 2.53 (m, 3H). MS: 420 [M+H] ⁺ .

Example 14. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(cyclobutylamino)butan- 2-enamide

The synthesis is carried out in the same manner as in Example 11, except that cyclobutylamine is used instead of isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.01 (s, 1H), 9.66 (s, 1H), 8.50 (s, 1H), 8.14 (d, J = 9.0 Hz, 1H), 7.78 (s, 2H), 7.45 (t, J = 7.5 Hz, 1H), 7.28 (t, J = 8.2 Hz, 1H), 6.89 (dt, J = 15.4, 5.3 Hz, 1H), 6.51 (d, J = 15.4 Hz, 1H), 3.29 (d, J = 5.3 Hz, 2H), 3.33(br, 1H), 3.20 (p, J = 7.6 Hz, 1H), 2.11 (q, J = 8.5, 7.9 Hz, 2H), 1.76- 1.55 (m, 4H). MS: 460 [M+H] ⁺ .

Example 15. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(isopropyl(methyl) Amino)but-2-enamide

The synthesis is carried out in the same manner as in Example 11, except that isopropylmethylamine is used instead of the isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.05 (s, 1H), 9.66 (s, 1H), 8.54 (s, 1H), 8.16 (d, J = 9.1 Hz, 1H), 7.87 – 7.75 ( m, 2H), 7.46 (s, 1H), 7.28 (t, J = 8.2 Hz, 1H), 6.83 (dt, J = 15.4, 5.7 Hz, 1H), 6.53 (dt, J = 15.4, 1.8 Hz, 1H ), 3.21 (dd, J = 5.8, 1.6 Hz, 2H), 2.83 (p, J = 6.6 Hz, 1H), 2.16 (s, 3H), 0.99 (d, J = 6.5 Hz, 6H). MS: 462 [M+H] ⁺ .

Example 16. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(cyclobutyl(methyl) Amino)but-2-enamide

The synthesis is carried out in the same manner as in Example 11, except that cyclobutylmethylamine is used instead of the isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.05 (s, 1H), 9.66 (s, 1H), 8.54 (s, 1H), 8.17 (d, J = 9.1 Hz, 1H), 7.88 – 7.76 ( m, 2H), 7.48 (s, 1H), 7.29 (t, J = 8.1 Hz, 1H), 6.84 (dt, J = 15.4, 5.9 Hz, 1H), 6.52 (d, J = 15.4 Hz, 1H), 3.09 – 3.02 (m, 2H), 2.93 – 2.81 (m, 1H), 2.06 (s, 3H), 1.99 (dd, J = 9.8, 7.2 Hz, 2H), 1.80 (tt, J = 11.2, 8.9 Hz, 2H), 1.61 (ddt, J = 18.1, 10.4, 8.1 Hz, 2H). MS: 474 [M+H] ⁺ .

Example 17. (E)-N-(5-chloro-4-((3-chloro-2-fluorophenyl)amino)quinazolin-6-yl)-4-(cyclopropyl(methyl) Amino)but-2-enamide

The synthesis is carried out in the same manner as in Example 11, except that cyclopropylmethylamine is used instead of isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.02 (s, 1H), 9.66 (s, 1H), 8.50 (s, 1H), 8.14 (s, 1H), 7.77 (s, 2H), 7.48 – 7.42 (m, 1H), 7.28 (s, 1H), 6.85 (s, 1H), 6.49 (s, 1H), 4.11 (s, 1H), 3.16 (s, 2H), 2.29 (s, 3H), 0.46 (s, 2H), 0.35 (s, 2H). MS: 460 [M+H] ⁺ .

Example 18. (E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl)-4 -(Cyclopropyl(methyl)amino)but-2-enamide

Synthesis was carried out in the same manner as in Example 11, except that 3-chloro-4-(pyridin-2-ylmethoxy)aniline was used instead of the 3-chloro-2-fluoroaniline in step 1), and 3-chloro-2-fluoroaniline was used. Propylmethylamine was used instead of isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.00 (s, 1H), 9.57 (s, 1H), 8.60 (ddd, J = 4.8, 1.8, 0.9 Hz, 1H), 8.52 (s, 1H), 8.12 (d, J = 9.0 Hz, 1H), 7.94 – 7.84 (m, 2H), 7.75 (d, J = 9.0 Hz, 1H), 7.62 – 7.54 (m, 2H), 7.38 (ddd, J = 7.6, 4.8, 1.2 Hz, 1H), 7.27 (d, J = 9.0 Hz, 1H), 6.86 (dt, J = 15.4, 6.1 Hz, 1H), 6.49 (dt, J = 15.4, 1.6 Hz, 1H), 5.31 ( s, 2H), 3.38 – 3.30 (m, 2H), 2.29 (s, 3H), 1.77 (tt, J = 6.6, 3.6 Hz, 1H), 0.46 (dt, J = 6.1, 3.0 Hz, 2H), 0.39 – 0.31 (m, 2H). MS: 549 [M+H] ⁺ .

Example 19. (E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl)-4 -(Cyclobutyl(methyl)amino)but-2-enamide

Synthesis was carried out in the same manner as in Example 11, except that 3-chloro-4-(pyridin-2-ylmethoxy)aniline was used instead of the 3-chloro-2-fluoroaniline in step 1), and 3-chloro-2-fluoroaniline was used. Butylmethylamine was used instead of isopropylamine in step 4) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.01 (s, 1H), 9.57 (s, 1H), 8.60 (dt, J = 4.9, 1.3 Hz, 1H), 8.52 (s, 1H), 8.12 ( d, J = 9.0 Hz, 1H), 7.94 – 7.84 (m, 2H), 7.74 (d, J = 9.0 Hz, 1H), 7.62 – 7.55 (m, 2H), 7.37 (ddd, J = 7.7, 4.8, 1.2 Hz, 1H), 7.27 (d, J = 9.0 Hz, 1H), 6.83 (dt, J = 15.4, 6.0 Hz, 1H), 6.52 (dt, J = 15.4, 1.8 Hz, 1H), 5.31(s, 2H), 3.05 (dd, J = 6.0, 1.6 Hz, 2H), 2.93 – 2.78 (m, 1H), 2.06 (s, 3H), 2.03 – 1.95 (m, 2H), 1.87 – 1.72 (m, 2H) , 1.60 (tdd, J = 15.0, 7.0, 4.9 Hz, 2H). MS: 563 [M+H] ⁺ .

Example 20. (R,E)-N-(5-chloro-4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)quinazolin-6-yl) -3-(1-methylpyrrolidin-2-yl)acrylamide

Synthesis is carried out in the same manner as in Example 11, except that 3-chloro-4-(pyridin-2-ylmethoxy)aniline is used instead of the 3-chloro-2-fluoroaniline of step 1), and ( R,E)-3-(1-methylpyrrolidin-2-yl)acrylyl chloride replaces the bromocrotonyl chloride in step 3) for reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.00 (s, 1H), 9.57 (s, 1H), 8.60 (d, J = 4.8 Hz, 1H), 8.52 (s, 1H), 8.13 (d, J = 9.0 Hz, 1H), 7.89 (dd, J = 14.2, 5.4 Hz, 2H), 7.75 (d, J = 9.0 Hz, 1H), 7.58 (d, J = 8.2 Hz, 2H), 7.37 (t, J = 6.1 Hz, 1H), 7.27 (d, J = 8.9 Hz, 1H), 6.72 (dd, J = 15.3, 7.5 Hz, 1H), 6.52 (d, J = 15.3 Hz, 1H), 5.31 (s, 2H), 3.04 (s, 1H), 2.79 (q, J = 8.0 Hz, 1H), 2.22 (s, 3H), 2.18 (d, J = 9.2 Hz, 1H), 2.02 (dq, J = 14.4, 8.3 , 7.8 Hz, 1H), 1.74 (q, J = 8.4 Hz, 2H), 1.61 (d, J = 18.1 Hz, 1H). MS: 549 [M+H] ⁺ .

Example 21. (E)-N-(5-chloro-4-((3-chloro-2,4-difluorophenyl)amino)quinazolin-6-yl)-4-(dimethylamino) But-2-enamide

Synthesis was carried out in the same manner as in Example 11, except that 2,4-difluoro-3-chloroaniline was used to replace the 3-chloro-2-fluoroaniline in step 1) and dimethylamine hydrochloride was used. The isopropylamine of step 4) is reacted;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.02 (s, 1H), 9.58 (s, 1H), 8.50 (s, 1H), 8.17 (d, J = 9.0 Hz, 1H), 7.78 (s, 2H), 7.40 (s, 1H), 6.83 (dt, J = 15.5, 5.8 Hz, 1H), 6.53 (dt, J = 15.5, 1.7 Hz, 1H), 3.10 (dd, J = 5.9, 1.6 Hz, 2H ), 2.21 (s, 6H). MS: 452 [M+H] ⁺ .

Example 22. (E)-N-(5-chloro-4-((3-chloro-2,4-difluorophenyl)amino)quinazolin-6-yl)-4-(isopropylamino )But-2-enamide

The synthesis is carried out in the same manner as in Example 11, except that 2,4-difluoro-3-chloroaniline is used instead of the 3-chloro-2-fluoroaniline in step 1) for reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.04 (s, 1H), 9.59 (s, 1H), 8.46 (s, 1H), 8.13 (d, J = 9.0 Hz, 1H), 7.75 (d, J = 14.5 Hz, 2H), 7.44 – 7.34 (m, 1H), 6.91 (dt, J = 15.4, 5.3 Hz, 1H), 6.54 (dt, J = 15.4, 1.8 Hz, 1H), 3.43 (dd, J = 5.4, 1.8 Hz, 2H), 3.33(br, 1H), 2.82 (p, J = 6.2 Hz, 1H), 1.05 (d, J = 6.2 Hz, 6H). MS: 466 [M+H] ⁺ .

Example 23. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(dimethylamino)butan -Synthesis of 2-enamide

Step 1) 5-Methylquinazolin-4-ol

Add 2-amino-6-methyl-benzoic acid (4.53 g, 30 mmol) and formamidine acetate (3.12 g, 30.00 mmol) into ethanol (40 mL), heat to 80°C to reflux for 24 hours, and monitor by LCMS. After the reaction was completed, the system dropped to room temperature and a large amount of solid precipitated. Filter, and wash the filter cake with a small amount of petroleum ether. Collect and dry to obtain the product 5-methylquinazolin-4-ol (3.66 g, 22.85 mmol, yield 76.17%); MS: 161[M+H] ⁺ .

Step 2) 5-methyl-6-nitroquinazolin-4-ol

At room temperature, slowly add 5-methylquinazolin-4-ol (3.66 g, 22.85 mmol) to H ₂ SO ₄ (30 mL), lower the ice-salt bath to about -20°C, and add KNO in batches ₃ (2.54 g, 25.14 mmol) (about 20 minutes), and control the system temperature below -10°C. The system was slowly heated to 10°C and reacted for 2 hours, monitored by HPLC. After the reaction was completed, the system was slowly poured into crushed ice, a large amount of solids precipitated, filtered, and the filter cake was washed with water three times, collected, and dried to obtain the product 5-methyl-6-nitro. Quinazolin-4-ol (3.2 g, 15.60 mmol, yield 68.26%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.45 (s, 1H), 8.14 (d, J = 8.9 Hz, 1H), 7.92(s, 1H) 7.63 (d, J = 8.9 Hz, 1H), 2.77 (s, 3H).

Step 3) N-(3-chloro-2-fluorophenyl)-5-methyl-6-nitroquinazolin-4-amine

Suspend 5-methyl-6-nitroquinazolin-4-ol (1 g, 4.87 mmol) in triturous chloride (15 mL), stir at room temperature, add DMF (0.5 mL), and then heat the system to 100°C for reaction. After the system is clarified (about 3 hours), the reaction is refluxed for another 2 hours and monitored by LCMS (MeOH quenching system). After the reaction is completed, the system is directly concentrated under reduced pressure to obtain a brown solid; the solid obtained above (1 g , 4.47 mmol) suspended in 1,2-dichloromethane (15 mL), sonicated to disperse evenly, slowly add 3-chloro-2-fluoroaniline (2.60 g, 17.89 mmol) under ice bath conditions, and complete the dripping , remove the ice bath, heat to 50°C and react for 1 hour. LCMS monitors that the reaction is completed. The system is decompressed and the solvent is evaporated. MeOH is added to the residue, dispersed evenly by ultrasonic, filtered, and the filter cake is collected to obtain the product (990 mg, 2.98 mmol, collected rate 66.54%).

Step 4) N-(3-chloro-2-fluorophenyl)-5-methylquinazoline-4,6-diamine

Suspend N-(3-chloro-2-fluoro-phenyl)-5-methyl-6-nitro-quinolin-4-amino (990 mg, 2.98 mmol) in methanol (10 mL) and add Raney Ni (34.93 mg, 595.10 μmol), replaced with H ₂ atmosphere three times, stirred at room temperature for 30 min, the system gradually dissolved and became clear, and monitored by LCMS. After the reaction was completed, it was filtered through diatomaceous earth, and the filtrate was concentrated to obtain the product (890 mg, 2.94 mmol, yield 98.80%).

Step 5. (E)-4-Bromo-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)but-2-enamide

Add N-(3-chloro-2-fluorophenyl)-5-methylquinazoline-4,6-diamine (50 mg, 165.16 μmol) to NMP (3 ml), stir at room temperature, and add dropwise Add the acetonitrile solution of (E)-bromocrotonyl chloride (45 mg, 250 μmol) and react for 15 minutes. Monitor by LCMS. After the reaction is completed, the system is quenched with excess saturated sodium bicarbonate aqueous solution. The pH is adjusted to ≈ 8, and a large amount of solid is precipitated. , filtered, washed with ethyl acetate, and dried to obtain 700 mg of crude product for later use, MS: 449, 451 [M+H] ⁺ .

Step 6. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(dimethylamino)butan- 2-enamide

(E)-4-bromo-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)but-2-enamide (50 mg, 165.16 μmol) was added to DMF (3 mL). Under stirring at room temperature, dimethylamine hydrochloride and diisopropylethylamine were added respectively, heated to 50 degrees Celsius and reacted for 2 hours. LCMS monitored that the reaction was completed. The system was The excess saturated sodium bicarbonate aqueous solution was quenched, the pH was adjusted to ≈ 8, and a large amount of solid was precipitated, filtered, and purified with a solid preparation plate (DCM/MeOH=10:1) to obtain the target product (25 mg, 60.40 μmol, yield 36.57%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.76 (s, 1H), 8.89 (s, 1H), 8.41 (s, 1H), 7.69 (br, 2H), 7.19 (br, 3H), 6.76 ( dt, J = 15.4, 5.8 Hz, 1H), 6.40 (d, J = 15.5 Hz, 1H), 3.08 (d, J = 5.9 Hz, 2H), 2.72 (s, 3H), 2.19 (s, 6H). MS: 414 [M+H] ⁺ .

Example 24. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(isopropylamino)butan -2-enamide

The synthesis is carried out in the same manner as in Example 23, except that isopropylamine is used instead of the dimethylamine hydrochloride in step 6) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.73 (s, 1H), 8.91 (s, 1H), 8.48-8.44 (m, 1H), 7.81 – 7.51 (m, 2H), 7.40 – 7.02 (m , 3H), 6.90 – 6.78 (m, 1H), 6.40 (d, J = 16.2 Hz, 1H), 3.33(br, 1H), 2.82-2.79 (m, 4H), 2.71 (d, J = 3.6 Hz, 2H), 1.02 (d, J = 6.2 Hz, 6H). MS: 428 [M+H] ⁺ .

Example 25. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(cyclopropylamino)butan -2-enamide

The synthesis is carried out in the same manner as in Example 23, except that cyclopropylamine is used instead of step 6) dimethylamine hydrochloride for reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.73 (s, 1H), 8.06 (s, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.39 (s, 2H), 7.33 – 7.12 ( m, 3H), 6.85 (dt, J = 15.4, 5.1 Hz, 1H), 6.39 (d, J = 15.4 Hz, 1H), 3.38 – 3.33 (m, 4H), 2.80 – 2.67 (s, 3H), 1.01 (d, J = 6.2 Hz, 4H). MS: 426 [M+H] ⁺ .

Example 26. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(methylamino)butan-2 -enamide

The synthesis is carried out in the same manner as in Example 23, except that methylamine hydrochloride is used instead of step 6) dimethylamine hydrochloride for reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.83 (s, 1H), 8.25 (s, 1H), 7.64 (br, 3H), 7.19 (s, 3H), 6.80 (d, J = 15.6 Hz, 1H), 6.43 (d, J = 15.5 Hz, 1H), 3.45 (br, 3H), 2.72 (s, 3H), 2.40 (s, 3H). MS: 400[M+H] ⁺ .

Example 27. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(cyclobutylamino)butan -2-enamide

The synthesis is carried out in the same manner as in Example 23, except that cyclobutylamine is used instead of step 6) dimethylamine hydrochloride for reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.74 (s, 1H), 8.91 (s, 1H), 8.38 (br, 1H), 7.64 (s, 2H), 7.43 – 6.90 (m, 3H), 6.82 (dt, J = 15.6, 5.3 Hz, 1H), 6.39 (d, J = 15.4 Hz, 1H), 3.33-3.27 (m, 3H), 3.20 (d, J = 7.7 Hz, 1H), 2.72 (s , 3H), 2.11 (q, J = 8.2, 5.8 Hz, 2H), 1.79 – 1.48 (m, 4H). MS: 440 [M+H] ⁺ .

Example 28. (E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(isopropyl(methyl) )Amino)but-2-enamide

Synthesis was carried out in the same manner as in Example 23, except that isopropylmethylamine was used instead of step 6) dimethylamine hydrochloride for reaction; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.95 -9.66 (m, 1H), 8.91-8.45 (m, 1H), 7.84 (d, J = 23.3 Hz, 1H), 7.68 – 7.58 (m, 2H), 7.36 – 6.99 (m, 3H), 6.75 (d , J = 15.2 Hz, 1H), 6.39 (d, J = 17.9 Hz, 1H), 3.20 (d, J = 5.7 Hz, 2H), 2.83 (q, J = 6.5 Hz, 1H), 2.72 (d, J = 22.2 Hz, 3H), 2.16 (s, 3H), 0.99 (d, J = 6.5 Hz, 6H). MS: 442 [M+H] ⁺ .

Example 29. (R,E)-N-(4-((3-chloro-2-fluorophenyl)amino)-5-methylquinazolin-6-yl)-3-(1-methyl Pyrrolidin-2-yl)acrylamide

Synthesis was carried out in the same manner as in Example 23, except that (R,E)-3-(1-methylpyrrolidin-2-yl)acrylyl chloride was used to replace the bromocrotonyl chloride in step 5). to react;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.65 (s, 1H) 8.46 (s, 1H), 7.90 – 7.80 (m, 1H), 7.61 – 7.46 (m, 2H), 7.36-6.99 (m, 3H), 6.66 (td, J = 19.5, 17.8, 7.4 Hz, 1H), 6.39 (dd, J = 27.3, 15.4 Hz, 1H), 3.03 (d, J = 8.5 Hz, 1H), 2.26 – 2.13 (m ,6H), 2.00 (d, J = 11.3 Hz, 2H), 1.77 (td, J = 6.6, 6.0, 2.8 Hz, 2H), 1.59 (s, 2H). MS: 440 [M+H] ⁺ .

Example 30. (E)-N-(4-((3-chloro-2,4-difluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(dimethylamino )But-2-enamide

Synthesize in the same manner as in Example 23, except that 3-chloro-2,4-difluoroaniline is used instead of the 3-chloro-2-fluoroaniline in step 3) to react to obtain; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.96 (s, 1H), 9.74 (s, 1H), 8.86 (s, 1H), 8.41 (s, 1H), 7.78 (s, 1H), 7.62 (s, 1H), 6.99 (s, 1H), 6.80 – 6.71 (m, 1H), 6.38 (d, J = 15.4 Hz, 1H), 3.08 (d, J = 5.9 Hz, 2H), 2.71 (s, 3H), 2.19 (s , 6H). MS: 432 [M+H] ⁺ .

Example 31. (E)-N-(4-((3-chloro-4-fluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(dimethylamino)butan- 2-enamide

The synthesis is carried out in the same manner as in Example 23, except that 3-chloro-4-fluoroaniline is used instead of the 3-chloro-2-fluoroaniline in step 3) for the reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.90 (s, 1H), 8.99 (s, 1H), 8.51 (s, 1H), 8.02 (dd, J = 6.9, 2.6 Hz, 1H), 7.83 ( d, J = 8.8 Hz, 1H), 7.72 – 7.59 (m, 2H), 7.43 (t, J = 9.1 Hz, 1H), 6.78 (dt, J = 15.4, 5.9 Hz, 1H), 6.45 (d, J = 15.5 Hz, 1H), 3.12 – 3.06 (m, 2H), 2.72 (s, 3H), 2.20 (s, 6H). MS: 414 [M+H] ⁺ .

Example 32. (E)-N-(4-((3-chloro-2,4-difluorophenyl)amino)-5-methylquinazolin-6-yl)-4-(isopropyl Amino)but-2-enamide

Synthesis was carried out in the same manner as in Example 23, except that 3-chloro-4-fluoroaniline was used to replace the 3-chloro-2-fluoroaniline in step 3), and isopropylamine was used to replace the dimethylamine in step 6). Hydrochloride is obtained by reaction;

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.91 (s, 1H), 8.33 (s, 1H), 7.65 (br, 2H), 7.28 (br, 3H), 6.84 (dt, J = 15.4, 5.7 Hz, 1H), 6.48 (d, J = 15.4 Hz, 1H), 3.53 (d, J = 5.7 Hz, 2H), 3.33(br, 1H), 2.96 (q, J = 6.3 Hz, 1H), 2.72 ( s, 3H), 1.11 (d, J = 6.3 Hz, 6H). MS: 446 [M+H] ⁺ .

Experimental Example 1. Test of small molecule compounds inhibiting EGFR ^WT and HER2 kinase activities

Reagents and consumables: ULightTM-labeled Ploy GT Peptide (Perkin Elmer, catalog number TRF-0100-M); ULightTM-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 (Carna Biosciences, catalog number 08-016); EGFR kinase (Carna Biosciences , catalog number 08-115); HEPES (GIBCO, catalog number 15630-080); EGTA (Sigma, catalog number 03777-10G); EDTA (Sigma, catalog number EDS-100G); MgCl ₂ (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 (Perkin Elmer, catalog number 607290) ;Multifunction plate reader (Perkin Elmer, catalog number Envision)

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 a 3-fold concentration gradient dilution, 11 gradients. When adding the drug, dilute it with buffer solution to a dilution of 4 times the final concentration.

HER2 kinase detection: Prepare buffer. Use the buffer to prepare 40 nM 4X Her2 kinase solution, 40 μM 4X ATP solution, and 400 nM 4×ULight ^TM -labeled Ploy GT Peptide substrate solution. After the configuration is completed, mix the enzyme with the pre-diluted compounds of different concentrations and leave them at room temperature for 5 minutes. Set duplicate wells for each concentration. Add the corresponding substrate and ATP and react at room temperature for 120 minutes (set a negative and positive control). After the reaction is completed, add PT66 detection antibody, incubate at room temperature for 60 minutes and then detect with Envision.

EGFR ^WT kinase detection: Prepare buffer, use buffer to prepare 3.48 nM 4X EGFR kinase solution, 600 μM 4X ATP solution, 400 nM 4×ULight ^TM -labeled JAK-1 (Try1023) Peptide substrate solution. After the configuration is completed, mix the enzyme with the pre-diluted compounds of different concentrations and leave them at room temperature for 5 minutes. Set duplicate wells for each concentration. Add the corresponding substrate and ATP and react at room temperature for 120 minutes (set a negative and positive control). After the reaction is completed, add PT66 detection antibody, incubate at room temperature for 60 minutes and then detect with Envision.

Data calculation: Use Excel table to calculate well reading value and inhibition rate, well reading value = 10000* (well EU665 value) / (well EU615 value), inhibition rate = [(positive control well reading value - experimental well reading value)/( Positive control well reading - Negative control well reading)]*100%. Enter the compound concentration and corresponding inhibition rate into GraphPad Prism to calculate the IC ₅₀ value.

The test in Table 2 shows that the compounds of the present application can inhibit the activity of EGFR ^WT and HER2 tyrosine kinase, especially some of the compounds show strong inhibitory effects. The test results are summarized in Table 2 below.

Table 2 lists the results of the determination of the inhibitory activity of some compounds in this application against EGFR ^WT and HER2 tyrosine kinase, where A indicates that the IC ₅₀ is less than or equal to 50 nM, and B indicates that the IC ₅₀ is greater than 50 nM but less than or equal to 500 nM. C indicates that the IC ₅₀ is greater than 500 nM but less than or equal to 5000 nM, D indicates that the IC ₅₀ is greater than 5000 nM, and NT indicates that there are no relevant results.

Table 2 Results of determination of the inhibitory activity of the compounds of the present invention on EGFR and HER2 kinase Example EGFR ^WT HER2 Example EGFR ^WT HER2 No. IC ₅₀ nM IC ₅₀ nM No. IC ₅₀ nM IC ₅₀ nM 1 A A 17 A B 2 A B 18 A A 3 A C 19 A A 4 A A 20 A A 5 A A twenty one A A 6 A A twenty two A A 7 A A twenty three A C 8 A A twenty four A C 9 A B 25 A C 10 A A 26 A C 11 A B 27 A C 12 A B 28 A C 13 A B 29 A B 14 A B 30 A C 15 A A 31 A B 16 A B 32 A C

Experimental Example 2. Test of small molecule compounds inhibiting cell proliferation

This application uses the CCK8 method to detect the in vitro anti-proliferative activity of the compounds of the present invention on BT474, NCI-N87, HCC-827 and Ba/F3 EGFRvIII cell lines cultured in vitro.

Reagents and consumables: RPMI1640 (ThermoFisher, catalog number C11875500BT); DMEM (ThermoFisher, catalog number C11995500BT); Fetal calf serum (Hyclone, catalog number SV30087.03); 0.25% trypsin-EDTA (ThermoFisher, catalog number 25200072); Penicillin-Chain Hyclone (Cat. No. SV30010); DSMO (Life Science, Cat. No. 0231-500ML); CCK8 test kit (Dojindo, Cat. No. CK04-100); 96-well plate (Corning, Cat. No. 3599); multi-read Trigger (Perkin Elmer, catalog number Envision)

Cell lines: BT474 (from the Cell Bank of the Chinese Academy of Sciences), NCI-N87 (from ATCC) and HCC-827 (from ATCC), Ba/F3 EGFRvIII from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd.; among them, BT474, NCI -N87 and Ba/F3 EGFRvIII were cultured in RPMI1640 medium containing 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin. HCC-827 was cultured in 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin. Culture in DMEM medium with μg/mL streptomycin.

Specific experimental methods:

1. Dissolve the test compound in DSMO to form a stock solution and carry out gradient dilution, and then dilute it with the corresponding culture medium to obtain a 20-fold working concentration solution.

2. Dilute the cells in the logarithmic growth phase with culture medium to adjust to a specific cell concentration, and add 80 μL of cell suspension to the 96-well plate so that the cell densities of BT474, NCI-N87, HCC-827 and Ba/F3 EGFRvIII are respectively For 10000 cells/well, 8000 cells/well, 5000 cells/well and 8000 cells/well. Place it in a 37°C, 5% carbon dioxide gas incubator and culture overnight. Among them, Ba/F3 EGFRvIII cells directly enter the next step of drug treatment, while BT474, NCI-N87, and HCC-827 need to be cultured overnight and then add drug treatment.

3. Add 20 μL drug solution to each well of the 96-well plate in which cells have been seeded. The highest concentration of the tested compound is 10 μM, 10 concentrations, 4-fold gradient dilution, and duplicate wells. At the same time, a control group without drugs was set up.

4. After the cells were cultured for 72 hours, CCK8 was used to detect cell viability. Use GraphPad Prism software to create dose-response curves and calculate IC ₅₀ .

Table 3 lists the results of the anti-proliferative activity of representative compounds in the present invention on BT474, NCI-N87, HCC-827 and Ba/F3 EGFRvIII cells. Among them, A means that the IC ₅₀ is less than or equal to 50 nM, B means that the IC ₅₀ is greater than 50 nM but less than or equal to 500 nM, C means that the IC ₅₀ is greater than 500 nM but less than or equal to 5000 nM, D means that the IC ₅₀ is greater than 5000 nM, and NT means not There are relevant results.

Table 3. Measurement results of anti-proliferative activity of representative compounds of the present invention on BT474, NCI-N87, HCC-827 and Ba/F3 EGFRvIII cells Example hBT474 htK HCC-827 Ba/F3EGFRvIII No. IC50 IC50 IC50 IC50 1 A A B A 2 NT NT A A 3 C B A A 4 B B NT A 5 A A C A 6 A A C A 7 NT NT A A 8 NT A B A 9 NT NT A A 10 NT NT A A 11 NT NT A A 12 NT NT A A 13 NT NT A A 14 NT NT A A 15 NT NT A A 16 NT NT A A 17 NT NT A A 18 NT A B A 19 NT A B A 20 NT A B A twenty one NT NT A A twenty two NT NT A A twenty three NT NT A A twenty four NT NT A A 25 NT NT A A 26 NT NT A A 27 NT NT A A 28 NT NT A A 29 NT NT A A 30 NT NT A A 31 NT NT A A 32 NT NT A A

The results in Table 3 show that the above compounds of the present application showed excellent to good anti-tumor proliferation activity against BT474, NCI-N87 and Ba/F3 EGFRvIII cells. Moreover, the compound of the present application also showed excellent anti-proliferation activity on HCC-827 cell line.

Experimental Example 3. Pharmacokinetic test of small molecule compounds

In this experiment, after a single oral administration and intravenous injection of some compounds of the present application were administered to SD rats, the pharmacokinetic characteristics of the compounds of the present application were studied, and the ability of the compounds of the present application to penetrate the blood-brain barrier was studied. At the same time, PYROTINIB and NERATINIB were tested accordingly and compared with the compounds of the present application.

(1) Reagents, instruments and animals used

Table 4. Test reagents Reagents supplier Item number Batch number DMSO Innochem D3851 INAH03010 Kolliphor®HS 15 (Solutol) SIGMA 42966 BCBZ6212 Acetonitrile Fisher 75-05-8 186342 Formic acid DIKMA 64-18-6 2486917 Ammonium acetate SIGMA 431311 MKCD5084 Ultrapure water homemade - -

Table 5. Test equipment instrument brand Model Liquid chromatography mass spectrometry (LC-MS) AB Sciex 5500Q-trap High speed refrigerated centrifuge Thermo ST40R One hundred thousandth scale Mettler XPE205 single hole pipette Eppendorf Research plus series Electric 12-hole volley gun Thermo Novus series

Table 6. Experimental rats Animals & strains: SD rat (male) Animal level: SPF level Animal source: Beijing Huafukang Biotechnology Co., Ltd. Weight range, age: 180g-350g on the first day of administration, 8 weeks (based on body weight)

(2) Sample preparation preparation

1. Intravenous injection (IV) group: Weigh an appropriate amount of the compound to be tested, completely dissolve it in an appropriate volume of solvent (DMSO/Solutol/H ₂ O=5/10/85 V/V, add 2 meq HCl), Stir, vortex and/or sonicate. After obtaining the solution, gradually increase the solvent to the final volume to reach the target concentration, vortex, and sonicate to obtain a uniform solution, which is filtered with a 0.22 μm PVDF filter.

2. Oral (PO) group: Weigh an appropriate amount of the compound to be tested and completely dissolve it in an appropriate volume of solvent (DMSO/Solutol/H ₂ O

=5/10/85 V/V, add 2 meq HCl), stir, vortex and/or sonicate. After obtaining the solution, gradually increase the solvent to the final volume to reach the target concentration, vortex, and sonicate to obtain a uniform solution.

(3) Administration and sampling of rats

The animals were randomly grouped according to their body weight. After grouping, the animals in each group had the same body weight (not exceeding ±20% of the average body weight). At the same time, the IV group did not fast, and the PO group fasted overnight (>12 hours) and were given food 2 hours after administration. All animals have free access to water. Tables 7 and 8 below provide the dosing regimen and pharmacokinetic sampling plan, respectively.

Table 7. Dosage regimen Group Dosing method Number of animals sample Dosage Dosing volume Preparation concentration (mg/kg) (mL/kg) (mg/mL) 1 IV 3 Whole blood 3 2 1.5 2 PO 3 Whole blood 10 10 1 3 PO 3 whole blood/brain 10 10 1

Table 8. Pharmacokinetic Sampling Plan Group Dosing method Sample collection anticoagulant Pharmacokinetic time points 1 IV Jugular Vein Cannulation (JVC) K2EDTA Whole blood: before dosing, 5, 15, 30 minutes, 1, 2, 4, 7 and 24 hours after dosing 2 PO Jugular Vein Cannulation (JVC) K ₂ EDTA Whole blood: before dosing, 15, 30 minutes, 1, 2, 4, 7 and 24 hours after dosing 3 PO Blood was drawn from the heart, animals were anesthetized, and brain tissue was collected. K ₂ EDTA Whole blood: 2 hours Brain tissue: 2 hours

Rats were administered drugs according to the above protocol, and blood and brain tissue samples were collected and processed at predetermined time points (collection and processing were carried out according to conventional methods in this field).

(4) Sample analysis

The brain was weighed and homogenized by adding 4 times the amount of ultrapure water. Add 6 times the volume of acetonitrile to the whole blood sample and brain homogenate respectively, vortex for 1 min, and centrifuge at 4°C and 4500 rpm for 15 min. The supernatant is diluted 2 times with ultrapure water, and the samples are analyzed by LC/MS.

(5) Data analysis:

Pharmacokinetic parameter calculations will be performed using WinNonlin software. If plasma drug concentration-time data are available, the following pharmacokinetic parameters will be calculated: CL (clearance); V _d (apparent volume of distribution); T _1/2 (elimination half-life); C _max (peak Concentration); T _max (time to peak); AUC (area under the plasma concentration-time curve); MRT (mean residence time); F% (bioavailability).

The test results are shown in Tables 9-15 below, which respectively provide the rat plasma concentrations of Compounds 1 and 2, 19 and 20, as well as pyrotinib and neratinib in the Examples of the present application at each time point, as well as each The pharmacokinetic parameter values also provide the concentrations and ratios of Compounds 1 and 2, 19 and 20, as well as pyrotinib and neratinib in the brain and blood of rats in the Examples of the present application. It can be seen from the above results that compounds 1 and 2, 19 and 20 of the present application all show excellent ability to penetrate the blood-brain barrier, which is far superior to the already marketed pyrotinib and nerlatinib. This also shows that the compound of the present application not only has excellent EGFR and HER2 kinase inhibitory activity, and can inhibit cell proliferation at the cellular level, but also has excellent ability to penetrate the blood-brain barrier, and is expected to be applied to EGFR and/or HER2 kinase mediated related diseases, especially those related to brain metastasis.

Table 9. Rat plasma concentrations of compounds 1 and 2 in the examples of this application Time point ( hr ) Example 1 Blood drug concentration ( ng/mL ) Example 2 Blood drug concentration ( ng/mL ) IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg 0 0 0 0 0 0.083 413±49 / 917±144 / 0.25 303±51 15.9±4 567±52 58.1 ± 22.3 0.5 222±45 49.6 ± 11.5 420±18 203±41 1 138±28 75.9 ± 5.4 308±16 362±18 2 73.1 ± 12.5 98.8 ± 4.8 149±8 348±72 4 23.3 ± 2.5 86.1 ± 26.8 41.3 ± 8.8 124±13 7 8.49 ± 2.19 78.1 ± 20.2 12±1.8 82.8 ± 25.4 twenty four 1.24±NA 1.83 ± 0.31 2.04±0.2 3.51 ± 0.88

Table 10. Rat pharmacokinetic parameters of the compounds of Examples 1 and 2 of the present application parameters Example 1 Example 2 IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg T _max (hr) / 2.67 ± 1.15 / 1.33 ± 0.58 C _max (ng/mL) / 103±8 / 378 ± 43 T _1/2 (hr) 4.00 ± 1.99 3.15 ± 0.10 5.18 ± 0.29 3.87 ± 0.50 AUC _0-t (hr*ng/mL) 552±28 1240±201 1133±58 2051±377 AUC _inf (hr*ng/mL) 567±37 1248 ± 203 1148±59 2071±369 _{Cl_obs} (mL/min/kg) 88.5 ± 5.8 / 43.6 ± 2.3 / V _d (L/kg) 31.2 ± 16.6 / 19.6±2.0 / F% / 66.1 ± 10.7 / 54.1 ± 9.6

Table 11. Rat plasma concentrations of compounds 19 and 20 in the examples of this application Time point (hr) Example 19 Blood drug concentration ( ng/mL ) Example 20 Blood drug concentration ( ng/mL ) IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg 0 0 0 0 0 0.083 637 ± 164 / 691±52 / 0.25 478 ± 106 18.8±3 506±9 11.8 ± 2.7 0.5 364 ± 109 64.9 ± 17.6 388 ± 19 40.7 ± 14.4 1 312±81 145±67 240±20 67.2 ± 21.8 2 222±60 131±51 140±24 64.1±12 4 118±46 113±42 93.2 ± 11.9 67.5 ± 10.8 7 92.3 ± 27.5 152±84 42.9 ± 3.2 83.9 ± 23.1 twenty four 6.87 ± 1.88 40.3 ± 49.1 3.10±0.43 10.1±4.6

Table 12. Rat pharmacokinetic parameters of the compounds in Examples 19 and 20 of the present application parameters Example 19 Example 20 IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg T _max (hr) / 3.33 ± 3.21 / 5.00 ± 3.46 C _max (ng/mL) / 167±63 / 89.7 ± 14.0 T _1/2 (hr) 4.89 ± 0.93 5.09 ± 1.55 4.21 ± 0.28 5.94 ± 2.19 AUC _0-t (hr*ng/mL) 2191 ± 623 2477±658 1450±75 1257±161 AUC _inf (hr*ng/mL) 2241±611 2845±316 1469±73 1353±97 _{Cl_obs} (mL/min/kg) 23.3 ± 5.5 / 34.1 ± 1.7 / V _d (L/kg) 10.1±3.8 / 12.5 ± 1.5 / F% / 33.9 ± 9.0 / 26.0±3.3

Table 13. Rat plasma concentrations of pyrotinib and neratinib Time point (hr) Pyrotinib plasma concentration (ng/mL) Neratinib plasma concentration (ng/mL) IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg 0 0 0 0 0 0.083 5507 ± 664 / 1460±282 / 0.25 3933 ± 363 42.3 ± 26.2 986 ± 337 38.4 ± 13.1 0.5 3007±205 76.2 ± 48.5 732 ± 242 87.2 ± 26.4 1 1903±572 98.2 ± 83 395 ± 132 93.3 ± 23.8 2 1086±320 172±114 179±32.9 142±38 4 429±94 160±36 128±22.7 194±73.7 7 122±14 198±97.8 72.3 ± 16.1 64.4 ± 50.2 twenty four 6.06 ± 1.83 4.51 ± 1.98 5.22 ± 1.96 1.44 ± 0.45

Table 14. Pharmacokinetic parameters of pyrotinib and neratinib in rats parameters Pyrotinib Neratinib IV 3 mg/kg PO 10 mg/kg IV 3 mg/kg PO 10 mg/kg T _max (hr) / 4.33 ± 2.52 / 2.67 ± 1.15 C _max (ng/mL) / 246±76 / 201±66 T _1/2 (hr) 3.45 ± 0.24 3.60±0.37 4.38 ± 0.55 2.97 ± 0.27 AUC _0-t (hr*ng/mL) 8310±892 2789 ± 1109 2388 ± 267 1466 ± 637 AUC _inf (hr*ng/mL) 8340±890 2813 ± 1110 2422 ± 252 1471±638 _{Cl_obs} (mL/min/kg) 6.04 ± 0.63 / 20.8±2.0 / V _d (L/kg) 1.81 ± 0.26 / 7.94 ± 1.71 / F% / 10.1±4.0 / 18.2 ± 7.89

Table 15. Concentrations and ratios of compounds 1 and 2, 19 and 20, as well as pyrotinib and neratinib in the brain and whole blood of the examples of this application (PO 10 mg/kg, sampling time, 2 hours after administration) Example Blood drug concentration (ng/mL) Brain concentration (ng/g) brain / blood ratio Pyrotinib 884 25.8 0.0291 Neratinib 440 14.4 0.0366 1 256 373 1.46 2 616 3168 5.45 19 446 788 1.77 20 109 116 1.06

The results in Table 9 and Table 10 show that Example 1 and Example 2 have excellent pharmacokinetic parameters and are suitable for the development of oral inhibitors. The results in Table 15 show that both Example 1 and Example 2 have extremely strong properties of penetrating the blood-brain barrier and are suitable for the treatment of primary brain tumors and the treatment of brain metastases. The results in Tables 11-12 and 15 also show that Examples 19 and 20 also have excellent pharmacokinetic properties and strong ability to penetrate the blood-brain barrier. Based on the results in Table 3, Table 9, Table 10, Tables 11-12 and Table 15, it can be seen that the compound of the present invention is expected to be developed into a therapeutic drug for glioma.

To sum up, the compounds in this application all show excellent inhibitory activity against EGFR kinase, and also show good to excellent inhibitory activity against HER2 kinase. In terms of cells, all compounds in this application show excellent inhibitory activity against Ba/F3 EGFRvIII cell line. Excellent anti-proliferation activity; at the same time, pharmacokinetic tests also found that the compound of the present application exhibits excellent ability to penetrate the blood-brain barrier (much better than existing drugs on the market). Therefore, it is especially suitable for EGFRvIII-induced tumors such as glia. tumors or EGFR/HER2-driven tumors that metastasize to the brain, the compound of the present application is expected to become a therapeutic drug for the above diseases.

Approved quinazoline drugs such as Gefitinib, Erlotinib, Icotinib, Afatinib and Lapatinib are Inability to effectively penetrate the blood-brain barrier. At the same time, most of the current compounds with quinazoline as the core are substituted at the 6- and 7-positions of the quinazoline ring, and almost none are substituted at the 5-position. The applicant's research found that in the quinazoline ring An allylamide group is introduced at the 6-position of the ring, and a halogen (such as Cl) or alkyl (such as methyl) substitution is introduced at the 5-position. The allylamide group can form an irreversible combination with the EGFR or HER2 target. Covalent binding, and the halogen or alkyl group introduced at the 5-position can lock the orientation of the allylamide group, which is more conducive to the covalent binding of allylamide to the EGFR or HER2 target. This design not only realizes the strong covalent binding ability of the compound of the present invention to the EGFR or HER2 target, but also greatly improves the blood-brain penetration ability of the compound of the present invention.

The above is the preferred embodiment of the present invention. It should be noted that for those with ordinary knowledge in the technical field to which the invention belongs, several improvements can be made to the embodiment of the present invention without departing from the principles described in the present invention. and modifications, these improvements and modifications should also be regarded as the protection scope of the present invention.

without

without.

Claims (18)

A compound represented by formula (I), its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof,

In formula (I), m is 0, 1 or 2; A is halogen, C ₁ -C ₃ alkyl; Z is NH or O; R ₁ is hydrogen, hydroxyl, 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkyl substituted with hydroxyl, C _{1 -} C ₃ C ₁ -C ₆ alkyl substituted by alkoxy, or C ₁ -C ₆ alkyl substituted by C _{3 -} C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group contains 1-2 A heteroalicyclic group with a heteroatom selected from N, O or S, which is unsubstituted or substituted by a C ₁ -C ₃ alkyl group, a C ₁ -C ₄ alkyl group, a hydroxyl group, a cyano group, One or both of aminocarboxylic acid group, mono- or disubstituted C ₁ -C ₃ aminocarboxylic acid group, C _{1 -} C ₃ alkyl styrene group, C _{1 -} C ₃ alkyl styrene group, oxo (=O) species substitution; R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are each independently hydrogen, halogen, C ₁ -C ₆ alkyl, -O-(CH ₂ )nR ₇ , R ₇ is hydrogen, C ₁ -C ₃ alkyl, 1 to 3 selected from halogen, cyano, hydroxyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, halogenated C ₁ -C ₃ alkyl, C ₃ - Aryl or heteroaryl substituted or unsubstituted by the substituents in C ₄ cycloalkyl, C ₂ -C ₃ alkynyl, C ₂ -C ₃ alkenyl or -NR'R", n is 0-3 Integer, the aryl group is a monocyclic or bicyclic group containing 6 to 12 carbon ring atoms and at least one aromatic ring, and the heteroaryl group is a heteroatom containing 1-3 selected from N, O, and S. A monocyclic or bicyclic group containing 5 to 10 ring atoms, R' and R" are each independently H or a C ₁ -C ₃ alkyl group. The compound of claim 1, its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof, wherein the compound has the following general formula,

In formula (II), A is halogen, C ₁ -C ₃ alkyl; Z is NH or O; R ₁ is hydrogen, hydroxyl, 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a , R ^b is each independently hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkyl substituted by hydroxyl, C ₁ - substituted by C _{1 -} C ₃ alkoxy C ₆ alkyl, or C ₁ -C ₆ alkyl substituted by C ₃ - C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group contains 1-2 selected from N, O or S Heteroalicyclic groups of heteroatoms, which are unsubstituted or substituted by C ₁ -C ₃ alkyl, C ₁ -C ₄ alkyl acyl, hydroxy, cyano, amino acyl, mono- or disubstituted One or two substitutions of C ₁ -C ₃ aminocarboxyl group, C _{1 -} C ₃ alkyl prenyl group, C _{1 -} C ₃ alkyl prenyl group, and oxo (=O); R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen, halogen, C ₁ -C ₆ alkyl, -O-(CH ₂ )nR ₇ , and R ₇ is hydrogen, C ₁ -C ₃ alkyl, and 1 To 3 selected from halogen, cyano, hydroxyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, halogenated C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₃ alkynyl, C ₂ -C ₃ alkenyl or -NR'R" substituted or unsubstituted aryl or heteroaryl group, n is an integer of 0-3, and the aryl group contains A monocyclic or bicyclic group with 6 to 12 carbon ring atoms and at least one aromatic ring. The heteroaryl group contains 1 to 3 heteroatoms selected from N, O, and S as ring atoms and contains 5 to 10 A monocyclic or bicyclic group of ring atoms, R' and R" are each independently H or a C ₁ -C ₃ alkyl group. The compound described in claim 1 or 2, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein A is Cl, F or methyl; Z is NH. The compound as described in claim 1 or 2, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein R ₁ is a 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkyl substituted by hydroxyl, C ₁ -C ₃ alkyl substituted by C _{1 -} C ₃ alkoxy; so The 4-7-membered heteroalicyclic group is pyrrolidinyl, piridinyl, pyrazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, and the above groups are unsubstituted or replaced by Methyl, ethyl, propyl, isopropyl, aldehyde, acetyl, propyl, hydroxyl, cyano, aminocarboxyl, methylpropyl, ethylpropyl, propylpropyl, isopropyl One or two substituted groups include methylprenylene, ethylprenylene, propylprenylene, isopropylprenylene, and oxo (=O). The compound of claim 4, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein R ₁ is pyrrolidin-1-yl, pyridin-1-yl, 1-methyl Pyrazin-4-yl, 1-ethylpyrazin-4-yl, morpholinyl, tetrahydrofuran 2-yl, tetrahydrofuran 3-yl, tetrahydropyran 2-yl, tetrahydropyran 3-yl, thio Morpholinyl, dimethylamino, diethylamino, dipropylamino, diisopropylamino, methylethylamino, methylpropylamino or ethylpropylamino. The compound of claim 5, its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof, wherein R ₁ is dimethylamino. The compound of claim 1, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein m is 0 or 1, and R ₁ is a 4-7 membered heteroalicyclic group or -NR ^a R ^b , R ^a and R ^b are each independently hydrogen, C ₁ -C ₃ alkyl, C ₃ -C ₆ cycloalkyl; the 4-7 membered heteroalicyclic group is pyrrolidinyl, pyridinyl, Pyrazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, and the above groups are unsubstituted or replaced by methyl, ethyl, propyl, isopropyl, aldehyde, acetyl group, propyl group, hydroxyl, cyano group, amino group, methyl group, ethyl group, propyl group, isopropyl group, methyl group, ethyl group, propyl group One or two kinds of substitutions are made from stylene, isopropyl stylene, and oxo (=O). The compound of claim 1, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein m is 0 or 1, R ₁ is 1-methyl-pyrrolidin-2-yl, 1-ethyl-pyrrolidin-2-yl, 1-isopropyl-pyrrolidin-2-yl, methylamino, ethylamino, propylamino, isopropylamino, cyclopropylamino, cyclobutyl Amino, methylisopropylamino, N-methyl-N-cyclopropylamino, N-methyl-N-cyclobutylamino, pyrrolidin-1-yl, pyridin-1-yl, 1-methyl pyrazin-4-yl, 1-ethylpyrazin-4-yl, morpholinyl, tetrahydrofuran 2-yl, tetrahydrofuran 3-yl, tetrahydropyran 2-yl, tetrahydropyran 3-yl, sulfide morpholinyl, dimethylamino, diethylamino, dipropylamino, diisopropylamino, methylethylamino, methylpropylamino or ethylpropylamino. The compound of claim 1 or 2, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen , fluorine, chlorine, methyl, ethyl, propyl, isopropyl, -O-(CH ₂ )nR ₇ , and at least 3 of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are hydrogen, R ₇ is hydrogen, methyl, ethyl, propyl, isopropyl or 1 to 3 selected from fluorine, chlorine, cyano, hydroxyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl A substituted or unsubstituted aryl or heteroaryl group substituted by a substituent in a base, fluoroethyl, trifluoromethyl, cyclopropyl, ethynyl, vinyl or -NR'R", n is an integer from 0 to 3 , R' and R" are each independently H or methyl, the aryl group is phenyl, and the heteroaryl group is pyridyl, pyrimidinyl, pyrrolyl, thienyl, furyl, or imidazolyl. The compound of claim 8, its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof, wherein R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen or fluorine. , chlorine, -O-(CH ₂ )nR ₇ , and at least 3 of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are hydrogen, and R ₇ is 1 to 3 selected from fluorine, chlorine, and cyanide. A substituted or unsubstituted aryl or heteroaryl group substituted by a substituent in a base, hydroxyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl, or trifluoromethyl group, n is 0 -3 is an integer, the aryl group is phenyl, and the heteroaryl group is pyridyl. The compound according to claim 10, its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof, wherein R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently hydrogen or fluorine. , chlorine, phenoxy, 2-fluorophenoxy, 3-fluorophenoxy, 4-fluorophenoxy, pyridin-2-ylmethoxy, pyridin-3-ylmethoxy, pyridine-4- Methoxy, 3-fluorobenzyloxy, 2-fluorobenzyloxy, 4-fluorobenzyloxy, 3-chlorobenzyloxy, 2-chlorobenzyloxy, 4-chlorobenzyl Oxygen group, and at least 3 of R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are hydrogen. The compound of claim 1 or 7, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein R ₂ , R ₃ , R ₅ , and R ₆ are each independently hydrogen, fluorine, Chlorine, methyl, ethyl, propyl, isopropyl, R ₄ is hydrogen, fluorine, chlorine, methyl, ethyl, propyl, isopropyl, -O-(CH ₂ )nR ₇ , and R ₂ , R ₃ , R ₄ , R ₅ , R ₆ at least 2 are hydrogen, R ₇ is hydrogen, methyl, ethyl, propyl, isopropyl or 1 to 3 selected from fluorine, chlorine, cyano , hydroxyl, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl, trifluoromethyl, cyclopropyl, ethynyl, vinyl or -NR'R" substituents Substituted or unsubstituted aryl or heteroaryl, n is an integer from 0 to 3, R' and R" are independently H or methyl, the aryl group is phenyl, and the heteroaryl group is pyridyl , pyrimidinyl, pyrrolyl, thienyl, furyl, imidazolyl. The compound of claim 12, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, wherein R ₂ , R ₃ , R ₅ , and R ₆ are each independently hydrogen, fluorine, or chlorine, R ₄ is hydrogen, fluorine, chlorine, phenoxy, 2-fluorophenoxy, 3-fluorophenoxy, 4-fluorophenoxy, pyridin-2-ylmethoxy, pyridin-3-ylmethoxy base, pyridin-4-ylmethoxy, 3-fluorobenzyloxy, 2-fluorobenzyloxy, 4-fluorobenzyloxy, 3-chlorobenzyloxy, 2-chlorobenzyloxy , 4-chlorobenzyloxy group, and at least 2 of R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are hydrogen. The compound described in claim 2, its stereoisomers, hydrates, and pharmaceutically acceptable salts thereof, the compound is selected from:

or

The compound described in claim 1 or 2, its stereoisomer, hydrate, and pharmaceutically acceptable salt thereof, said compound is selected from:

A pharmaceutical composition, including the compound as described in any one of claims 1 to 15, its pharmaceutically acceptable salts, stereoisomers, and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical composition of claim 16, wherein the pharmaceutical composition further contains one or more other therapeutic agents. A compound as described in any one of claims 1 to 15, its pharmaceutically acceptable salt, or stereoisomer in the preparation of drugs for the treatment of cancer and autoimmune diseases related to tyrosine kinase EGFR and HER2 Applications, wherein the cancer and autoimmune diseases include: fundus diseases, dry eye disease, 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 myeloid leukemia, non-Hodgkin lymphoma, Nasopharyngeal cancer, esophageal cancer, brain tumors, B-cell and T-cell lymphoma, lymphoma, multiple myeloma, biliary carcinosarcoma, cholangiocarcinoma.