CN106749267B

CN106749267B - Novel epidermal growth factor receptor inhibitors and uses thereof

Info

Publication number: CN106749267B
Application number: CN201611025615.5A
Authority: CN
Inventors: 王勇; 张小猛; 赵立文; 王小伟; 韩璐薇; 戴雪娟
Original assignee: Nanjing Sanhome Pharmaceutical Co Ltd
Current assignee: Nanjing Sanhome Pharmaceutical Co Ltd
Priority date: 2015-11-23
Filing date: 2016-11-22
Publication date: 2020-08-21
Anticipated expiration: 2036-11-22
Also published as: WO2017088746A1; CN106749267A; TW201718583A

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to a novel epidermal growth factor receptor inhibitor, a pharmaceutical composition containing the inhibitor, and application of the inhibitor or the pharmaceutical composition as a cancer prevention and/or treatment drug. The compounds show good EGFR inhibitory activity, and are expected to become drugs with specific curative effect and small side effect on drug-resistant tumors caused by EGFR mutation especially against mutated EGFR.

Description

Novel epidermal growth factor receptor inhibitors and uses thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a novel epidermal growth factor receptor inhibitor, a pharmaceutical composition containing the inhibitor, and application of the inhibitor or the pharmaceutical composition as a cancer treatment drug.

Background

Epidermal Growth Factor Receptor (EGFR) is an expression product of proto-oncogene C-erbB-1, and is one of EGFR family members. The EGFR family includes four members of EGFR (HER-1), ERBB2(HER-2), ERBB3(HER-3), and ERBB4 (HER-4). EGFR overexpression or mutation has been shown to generally trigger tumors. EGFR mutation causes the continuous activation of EGFR, the function of an autocrine loop is enhanced, a receptor down-regulation mechanism is damaged, and an abnormal signal conduction pathway is activated, thereby playing an important role in the evolution of tumors.

Treatment with EGFR-tyrosine kinase inhibitors (EGFR-TKI) against EGFR has become the gold standard in the field of non-small cell lung cancer treatment. However, clinical application shows that most patients have different drug resistance phenomena 6-12 months after the EGFR-TKI inhibitors such as gefitinib and erlotinib are treated, and further the curative effect of the drug is obviously reduced, and tumors progress. Research shows that the generation of EGFR-TKI resistance is related to secondary mutation of EGFR gene, wherein the most common mutation is mutation of No. 790 exon of EGFR gene, namely T790M gene mutation. Because methionine occupies a larger space than threonine, steric hindrance is formed, the affinity of ATP in an EGFR kinase region is changed, and the EGFR-TKI small-molecule drug cannot effectively block an EGFR activation signal, so that drug resistance is generated. Meanwhile, the first-generation EGFR inhibitor lacks selectivity of wild EGFR and mutant EGFR, and side effects such as rash, diarrhea and the like are commonly existed, so that the compliance of patients is influenced.

Therefore, the development of effective drugs for tumor patients with drug resistance caused by EGFR secondary mutation, particularly the development of drugs capable of further improving the selectivity of wild EGFR and mutant EGFR, enhancing the curative effect and reducing the side effect, has good application prospect.

Disclosure of Invention

The invention aims to provide an epidermal growth factor inhibitor with a general formula I or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, which shows good EGFR inhibitory activity, especially against mutated EGFR,

the invention also aims to provide the compound of the general formula II or the pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, the compound of the general formula I is prepared by taking the compound of the general formula II as a key intermediate, the reaction condition is mild, the yield and the purity are high,

the third purpose of the invention is to provide a compound of the general formula III or a salt thereof, the compound of the general formula I is prepared by taking the compound of the general formula III as a key intermediate, the reaction steps are few, the conditions are mild, the yield and the purity are high,

a fourth object of the present invention is to provide a method for preparing the compound of formula I, II or III of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof.

A fifth object of the present invention is to provide a pharmaceutical composition comprising the compound of formula I of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof and a pharmaceutically acceptable carrier, and a pharmaceutical composition comprising the compound of formula I of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof and another tumor suppressor.

The sixth object of the present invention is to provide the compound of the general formula I of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, and a method for treating and/or preventing cancer and use thereof in preparing a medicament for treating and/or preventing cancer.

Aiming at the above purpose, the invention provides the following technical scheme:

in a first aspect, the present invention provides a compound represented by the general formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof,

wherein the content of the first and second substances,

ring A is C₄-C₈Nitrogen-containing heterocyclic groups optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, when two or more substituents are present, two adjacent substituents can together with the atoms to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl;

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N;

R₁、R_2a、R_2b、R_3a、R_3band R_3cEach independently selected from the group consisting of hydrogen, halogen, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl and dialkylaminoacyl;

R₄、R₅and R₆Each independently selected from the group consisting of hydrogen, halogen, cyano, alkyl, alkoxy, and cycloalkyl, said alkyl, alkoxy, and cycloalkyl being optionally substituted with one or more of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, amino, monoalkylamino, dialkylamino, hydroxy, ammoniaArylalkyl, monoalkylaminoalkyl, dialkylaminoalkyl or hydroxyalkyl;

R₇selected from the group consisting of hydrogen, alkyl, cycloalkyl, and haloalkyl; and

R₈selected from the group consisting of hydrogen, alkyl and cycloalkyl, said alkyl and cycloalkyl being optionally substituted with one or more halogen, alkyl, haloalkyl, alkoxy, amino, monoalkylamino, dialkylamino or hydroxy groups; or

R₇、R₈And the N atom to which they are attached, form a nitrogen-containing heterocycloalkyl group that is optionally substituted with one or more halogen, alkyl, haloalkyl, alkoxy, amino, monoalkylamino, dialkylamino, hydroxy, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, or hydroxyalkyl groups.

In a second aspect, the present invention provides a compound of formula II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof,

wherein the ring A, X₁、X₂、X₃、X₄、R_2a、R_2b、R_3a、R_3b、R_3c、R₇And R₈Have the definition in the general formula I.

In a third aspect, the present invention provides a compound of formula III or a pharmaceutically acceptable salt thereof,

wherein the ring A, X₁、X₂、X₃、X₄、R_2a、R_2bHas the definition in the general formula I, and M is halogen.

In some embodiments, the present invention provides a compound of formula I, II or III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is C₄-C₈Nitrogen-containing heteromonocycloalkyl optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, and when there are two or more substituents, two adjacent substituents can together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl.

In some embodiments, the present invention provides compounds of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is C containing only one nitrogen atom₄-C₈Nitrogen-containing heteromonocycloalkyl optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, and when there are two or more substituents, two adjacent substituents can together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl.

In some preferred embodiments, the present invention provides a compound of formula I, II or III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is

n is selected from 1,2,3,4, 5 and 6, m is selected from 1,2,3 and 4, R_aSelected from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl, di-C_1-6Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-8Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent carbon atoms_aMay form C together with the carbon atom to which they are attached_3-8Cycloalkyl or C_3-8Heterocycloalkyl, or two R on the same carbon atom_aMay form C together with the carbon atoms to which they are jointly attached_3-8Cycloalkyl or C_3-8A heterocycloalkyl group.

n is selected from 1,2,3 and 4, m is selected from 1,2,3 and 4, R_aSelected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkyl amino acylTwo C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent carbon atoms_aMay form C together with the carbon atom to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or two R on the same carbon atom_aMay form C together with the carbon atoms to which they are jointly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group.

In other embodiments, the present invention provides compounds of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is C containing two nitrogen atoms₄-C₈Nitrogen-containing heteromonocycloalkyl optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, and when there are two or more substituents, two adjacent substituents can together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl.

m is selected from 1,2,3 and 4, p is selected from 0, 1 and 2, q is selected from 0, 1 and 2, R_aSelected from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, monoC_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl, di-C_1-6Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-8Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-8Cycloalkyl or C_3-8Heterocycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-8Cycloalkyl or C_3-8A heterocycloalkyl group.

m is selected from 1,2,3 and 4, p is selected from 0, 1 and 2, q is selected from 0, 1 and 2, R_aSelected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl, di-C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group.

In other embodiments, the present invention provides compounds of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is C containing one nitrogen atom and one oxygen atom₄-C₈Nitrogen-containing heteromonocycloalkyl optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, and when there are two or more substituents, two adjacent substituents can together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl.

m is selected from 1,2,3 and 4, p is selected from 0, 1 and 2, q is selected from 0, 1 and 2, R_aSelected from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl, di-C_1-6Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-8Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-8Cycloalkyl or C_3-8Heterocycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-8Cycloalkyl or C_3-8A heterocycloalkyl group.

m is selected from 1,2,3 and 4, p is selected from 0, 1 and 2, q is selected from 0, 1 and 2, R_aSelected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl, di-C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group. In some embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt thereofA salt, isomer, solvate, crystal or prodrug thereof, wherein ring A is C containing one nitrogen atom and one sulfur atom₄-C₈Nitrogen-containing heteromonocycloalkyl optionally substituted with one or more of halogen, oxo, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, monoalkylamino, dialkylamino, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, aminoacyl, monoalkylaminoacyl, dialkylaminoacyl, aryl, heteroaryl, and heterocycloalkyl, and when there are two or more substituents, two adjacent substituents can together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or two substituents attached to the same atom can form a substituted or unsubstituted cycloalkyl, heterocycloalkyl, or a substituted or unsubstituted heteroaryl.

In some embodiments, the present invention provides a compound of formula I, II or III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein X₁、X₂、X₃And X₄Are all C (R)₁)。

In some specific embodiments, the present invention provides compounds of the following formulae Ia, IIa and IIIa, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof,

wherein the ring A, R₁、R_2a、R_2b、R_3a、R_3b、R_3c、R₄、R₅、R₆、R₇、R₈And M has the definitions in formulas I, II and III above, n is an integer selected from 1,2,3 and 4.

In other embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein X₁、X₂、X₃And X₄At least one of which is N.

In other embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein X₁、X₂、X₃And X₄Two of which are N.

In other embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein X₁、X₂、X₃And X₄Three of which are N.

In other embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein X₁、X₂、X₃And X₄Are all N.

In some embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₁、R_2aAnd R_2bEach independently selected from hydrogen, halogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl and di-C_1-6An alkylaminoacyl group.

In some preferred embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein each R is₁、R_2aAnd R_2bEach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-6Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl and di-C_1-3An alkylaminoacyl group.

In other preferred embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₁、R_2aAnd R_2bEach independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethyl, trifluoroethyl, difluoromethyl, difluoroethyl, methoxy, ethoxy, propoxy, trifluoromethyloxy, nitro, cyano, hydroxy, amino, methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, N-methyl-N-ethylamino, N-methyl-N-propylamino, N-methyl-N-isopropylamino, N-ethyl-N-propylamino, N-ethyl-N-isopropylamino, N-propyl-N-isopropylamino, aminomethyl, aminoethyl, methyl, isopropyl, Aminopropyl, methylaminomethyl, methylaminoethyl, ethylaminomethyl, ethylaminoethyl, dimethylaminomethyl, dimethylaminoethyl, diethylaminomethyl, diethylaminoethyl, hydroxymethyl, hydroxyethyl, methoxymethyl, ethoxymethyl, propoxymethyl, methoxyethyl, and ethoxyethyl.

In other preferred embodiments, the present invention provides a compound of formula I, II or III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₁、R_2aAnd R_2bEach independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl,Isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, hydroxy, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, aminomethyl, aminoethyl, aminopropyl, methylaminomethyl, methylaminoethyl, ethylaminomethyl, ethylaminoethyl, dimethylaminomethyl, diethylaminomethyl, hydroxymethyl, and hydroxyethyl.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is_3a、R_3bAnd R_3cEach independently selected from hydrogen, halogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl and di-C_1-6An alkylaminoacyl group.

In other preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein each R_3a、R_3bAnd R_3cEach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-6Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl and di-C_1-3An alkylaminoacyl group.

In other preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is_3a、R_3bAnd R_3cEach independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethyl, trifluoroethyl, difluoromethyl, difluoroethyl, methoxy, ethoxy, propoxy, trifluoromethyloxy, nitro, cyano, hydroxy, amino, methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, N-methyl-N-ethylamino, N-methyl-N-propylamino, N-methyl-N-isopropylamino, N-ethyl-N-propylamino, N-ethyl-N-isopropylamino, N-propyl-N-isopropylamino, aminomethyl, aminoethyl, methyl, isopropyl, Aminopropyl, methylaminomethyl, methylaminoethyl, ethylaminomethyl, ethylaminoethyl, dimethylaminomethyl, dimethylaminoethyl, diethylaminomethyl, diethylaminoethyl, hydroxymethyl, hydroxyethyl, methoxymethyl, ethoxymethyl, propoxymethyl, methoxyethyl, and ethoxyethyl.

In other preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is_3a、R_3bAnd R_3cEach independently selected from hydrogen, fluoro, chloro, bromo, methyl, ethyl, propyl, isopropyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, hydroxy, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, aminomethyl, aminoethyl, aminopropyl, methylaminomethyl, methylaminoethyl, ethylaminomethyl, ethylaminoethyl, dimethylaminomethyl, diethylaminomethyl, hydroxymethyl, and hydroxyethyl.

In some embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇Selected from hydrogen, C_1-6Alkyl, aryl, heteroaryl, and heteroaryl,C_3-6Cycloalkyl and halo C_1-6An alkyl group.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇Selected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl and halo C_1-3An alkyl group.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇Selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and trifluoromethyl.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇Is methyl.

In some embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₈Selected from hydrogen, C_1-6Alkyl and C_3-6Cycloalkyl radical, said C_1-6Alkyl and C_3-6Cycloalkyl optionally substituted by one or more halogens, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino or hydroxy.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₈Selected from hydrogen, C_1-3Alkyl and C_3-6Cycloalkyl radical, said C_1-3Alkyl and C_3-6Cycloalkyl optionally substituted by one or more halogens, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino or hydroxy.

In some preferred embodiments, the present inventionCompounds of formula I or II or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof are provided, wherein R₈Selected from hydrogen, C_1-3Alkyl and C_3-6Cycloalkyl radical, said C_1-3Alkyl and C_3-6The cycloalkyl group is optionally substituted with one or more halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropyloxy, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, methylethylamino, methylpropylamino, ethylpropylamino, or hydroxy.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₈Selected from the group consisting of hydrogen, methyl, ethyl and propyl, said methyl, ethyl and propyl being optionally substituted with one or more amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, methylethylamino, methylpropylamino or ethylpropylamino groups.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₈Is ethyl substituted by dimethylamino.

In other embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇、R₈And the N to which they are attached form a four to eight membered azacycloalkyl group optionally substituted with one or more halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino, hydroxy, amino C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl or hydroxyalkyl.

In some preferred embodiments, the present inventionCompounds of formula I or II or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof are provided, wherein R₇、R₈And the N to which they are attached form a four to eight membered azacycloalkyl group optionally substituted with one or more halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino, hydroxy, amino C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl or hydroxyalkyl.

In some preferred embodiments, the present invention provides compounds of formula I or II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₇、R₈And the N to which they are attached form a five to six membered azacycloalkyl group optionally substituted with one or more halogens, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino, hydroxy, amino C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl or hydroxyalkyl.

In some embodiments, the present invention provides compounds of formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₄、R₅And R₆Each independently selected from hydrogen, halogen, cyano, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl radical, said C_1-6Alkyl radical, C_1-6Alkoxy and cycloalkyl optionally substituted by one or more halogens, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino, hydroxy, amino C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl and hydroxy C_1-6Alkyl substitution.

In some preferred embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₄、R₅And R₆Each independently selected from hydrogen, halogen, cyano, C_1-3Alkyl radical, C_1-3Alkoxy radical, C_3-6Cycloalkyl radical, said C_1-3Alkyl radical, C_1-3Alkoxy and C_3-6Cycloalkyl optionally substituted by one or more halogens, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino, hydroxy, amino C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl or hydroxy C_1-3Alkyl substitution.

In some preferred embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₄、R₅And R₆Are all hydrogen.

In some embodiments, the present invention provides a compound of formula III or a pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of fluorine, chlorine, bromine, and iodine.

In some preferred embodiments, the present invention provides a compound of formula III or a pharmaceutically acceptable salt thereof, wherein M is selected from the group consisting of chlorine, bromine, and iodine.

In some embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein

Ring A is

n is selected from 1,2,3 and 4;

m is selected from 1,2,3 and 4;

R_aselected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl, di-C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent carbon atoms_aMay form C together with the carbon atom to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or two R on the same carbon atom_aMay form C together with the carbon atoms to which they are jointly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group;

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N;

R₁、R_2a、R_2b、R_3a、R_3band R_3cEach independently selected from hydrogen, halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-6Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl and di-C_1-3An alkylaminoacyl group;

R₄、R₅and R₆Are all hydrogen;

R₇is methyl;

R₈is ethyl substituted by dimethylamino.

In other specific embodiments, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein

Ring A is

m is selected from 1,2,3 and 4;

p is selected from 0, 1 and 2;

q is selected from 0, 1 and 2; r_aSelected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl, di-C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group;

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N;

R₁、R_2a、R_2b、R_3a、R_3band R_3cEach independently selected from hydrogen, halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, monoC_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-6Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl and di-C_1-3An alkylaminoacyl group;

R₄、R₅and R₆Are all hydrogen;

R₇is methyl;

R₈is ethyl substituted by dimethylamino.

Ring A is

m is selected from 1,2,3 and 4;

p is selected from 0, 1 and 2;

q is selected from 0, 1 and 2; r_aSelected from hydrogen, C_1-3Alkyl radical, C_3-6Cycloalkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-3Alkylamino, di-C_1-3Alkylamino radical, amino radical C_1-3Alkyl, mono C_1-3Alkylamino radical C_1-3Alkyl, di-C_1-3Alkylamino radical C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, amino acyl, mono C_1-3Alkylaminoacyl, di-C_1-3Alkylaminoacyl radical, C_6-10Aryl radical, C_5-10Heteroaryl and C_3-6Heterocycloalkyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-6Cycloalkyl or C_3-6Heterocycloalkyl, or of the sameTwo R on one atom_aMay form C together with the atoms to which they are commonly attached_3-6Cycloalkyl or C_3-6A heterocycloalkyl group;

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N;

R₄、R₅and R₆Are all hydrogen;

R₇is methyl;

R₈is ethyl substituted by dimethylamino.

In some specific embodiments, the present invention provides the following compounds, or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof:

in other specific embodiments, the present invention provides the following compounds, or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof:

in a fourth aspect, the present invention provides a process for the preparation of compounds of formulae I, II and III of the present invention, comprising the steps of:

A) carrying out nucleophilic reaction on the raw material of the formula 1 and the raw material of the formula 2 to obtain an intermediate of a formula III;

B) reacting the intermediate of the formula III with the raw material of the formula 3 to prepare an intermediate of the formula 4;

C) carrying out nucleophilic reaction on the intermediate of the formula 4 to prepare an intermediate of a formula 5;

D) the intermediate of the formula 5 is subjected to reduction reaction to prepare an intermediate of a formula II;

E) the intermediate of formula II is reacted with the starting material of formula 6 to produce the compound of formula I.

The above ring A, X₁、X₂、X₃、X₄、R_2a、R_2b、R_3a、R_3b、R_3c、R₄、R₅、R₆、R₇、R₈And M has the meaning given in the formulae I, II, III.

In some embodiments, the present invention provides a method of making a compound of formula I of the present invention, comprising using a compound of formula II of the present invention.

In some embodiments, the present invention provides a method of making a compound of formula I of the present invention, comprising using a compound of formula III of the present invention.

In a fifth aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof and a pharmaceutically acceptable carrier.

In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of formula I, isomer, solvate, crystal or prodrug of the present invention, further comprising one or more compounds selected from the group consisting of: gefitinib, erlotinib, lapatinib, afatinib, vandetanib, canertinib, apatinib, dacatinib (dacomitinib), pelitinib (pelitinib), WZ4002, AG-490, AZD8931, AZD9291, and the like.

The compounds of formula I, isomers, solvates, crystals or prodrugs of the invention may be combined with pharmaceutically acceptable carriers to prepare pharmaceutical formulations suitable for oral or parenteral administration. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulations may be administered by any route, for example by infusion or bolus injection, by a route of absorption through epithelial or cutaneous mucosa (e.g. oral mucosa or rectum, etc.). Administration may be systemic or local. Examples of the formulation for oral administration include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions and the like. The formulations may be prepared by methods known in the art and include carriers conventionally used in the art of pharmaceutical formulation.

In a sixth aspect, the present invention provides a method for treating and/or preventing tumors by using the compound of the general formula I, isomer, solvate, crystal or prodrug of the present invention or the pharmaceutical composition of the present invention, and an application in preparing a medicament for treating and/or preventing tumors, which comprises administering the compound of the general formula I, isomer, solvate, crystal or prodrug of the present invention or the pharmaceutical composition comprising the compound of the general formula I, isomer, solvate, crystal or prodrug of the present invention to a population prone to tumor or a tumor patient, so as to effectively reduce the incidence of tumors and prolong the life of the tumor patient.

In some embodiments, the present invention provides a method of treating a drug resistant tumor by administering to a patient having a drug resistant tumor a therapeutically effective amount of a compound of formula I, isomer, solvate, crystal or prodrug of the present invention or a pharmaceutical composition comprising a compound of formula I, isomer, solvate, crystal or prodrug of the present invention.

In other embodiments, the present invention provides the use of a compound of formula I, isomer, solvate, crystal or prodrug of the present invention or a pharmaceutical composition of the present invention for the manufacture of a medicament for the treatment of a drug-resistant tumor. The drug resistant tumor may be a tumor that is resistant to multiple drugs, preferably to EGFR inhibitors, such as first, second, and third generation EGFR inhibitors, such as gefitinib, erlotinib, and lapatinib. The tumor includes but is not limited to solid tumor, preferably lung cancer, head and neck tumor, colorectal cancer, bladder cancer, pancreatic cancer, breast cancer, prostate cancer, stomach cancer, oral cancer, liver cancer, ovarian cancer. More preferably, the tumor is non-small cell lung cancer. In some embodiments, the present invention provides methods of treating drug resistant tumors with a compound of formula I, isomer, solvate, crystal or prodrug of the present invention, wherein said tumor carries an EGFR mutant gene. In one embodiment, the EGFR mutant gene carried by the tumor is the T790M mutation in exon 20. In another embodiment, the EGFR mutant gene carried by the tumor is a L858R mutation and/or a deletion/insertion mutation in exon 21. In another embodiment, the EGFR mutant gene carried by the tumor is a T790M and L858R double mutation. In other embodiments, the present invention provides a compound of formula I, isomer, solvate, crystal or prodrug of the present invention or a pharmaceutical composition of the present invention for use in the treatment of tumors, wherein the tumor treatment effect is manifested by a superior therapeutic effect, a high degree of selectivity and/or fewer side effects. In still other embodiments, the present invention provides a method of treating tumors by administering to a patient in need thereof a therapeutically effective amount of a compound of formula I, isomer, solvate, crystal or prodrug of the present invention or a pharmaceutical composition of the present invention, resulting in a tumor treatment effect manifested by a superior therapeutic effect, a high degree of selectivity and/or fewer side effects.

Definition of terms

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

"isomers" in the present invention include cis-trans isomers in either the cis or trans configuration, as well as enantiomers and diastereomers arising from chiral carbons.

The "pharmaceutically acceptable salt" of the present invention refers to a pharmaceutically acceptable salt of the compound of the present invention with an acid, which may be selected from: phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, citric acid, maleic acid, malonic acid, mandelic acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-toluenesulfonic acid, malic acid, methanesulfonic acid, and the like.

"solvate" of the present invention refers to a form of the compound of the present invention which forms a solid or liquid complex by coordination with a solvent molecule. Hydrates are a special form of solvates in which coordination occurs with water. Within the scope of the present invention, the solvate is preferably a hydrate.

The term "crystalline" as used herein refers to the various solid forms formed by the compounds of the present invention, including crystalline forms and amorphous forms.

The "prodrug" of the present invention refers to a compound which is converted into the compound of the present invention by reacting with an enzyme, gastric acid or the like under physiological conditions of an organism, that is, a compound which is converted into the compound of the present invention by oxidation, reduction, hydrolysis or the like by an enzyme and/or a compound which is converted into the compound of the present invention by hydrolysis or the like by gastric acid or the like.

The "pharmaceutical composition" of the present invention is intended to include a mixture of any one of the compounds described herein, including isomers, prodrugs, solvates, pharmaceutically acceptable salts, or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers.

The term "use in the preparation of a medicament for the treatment and/or prevention of a tumor" of the present invention means that the growth, development and/or metastasis of a tumor can be inhibited, and a therapeutically effective dose of a compound of the present invention is administered primarily to a human or animal in need thereof to inhibit, slow or reverse the growth, development or spread of a tumor in the subject.

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group, preferably a hydrocarbon group of 6 carbon atoms or less. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 2-methylbutyl and 2, 3-dimethylbutyl. The term "C_1-6Alkyl "refers to a straight or branched chain saturated hydrocarbon group containing 1 to 6 carbon atoms. The term "C_1-3Alkyl "refers to a straight or branched chain saturated hydrocarbon group containing 1 to 3 carbon atoms.

The "cycloalkyl group" in the present invention means a cyclic saturated, partially saturated hydrocarbon group, preferably a cycloalkyl group having 12 or less carbon atoms, more preferably a cycloalkyl group having 8 or less carbon atoms, and still more preferably a cycloalkyl group having 6 or less carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl. "C" of the invention_3-8Cycloalkyl "refers to a cyclic saturated hydrocarbon group containing 3 to 8 carbon atoms.

"alkoxy" in the context of the present invention means-O-alkyl.

"oxo" in the context of the present invention means O ═ e.g. formed by substitution of a carbon atom with an oxo group

The sulfur atom being substituted by an oxo group to form

The sulfur atom being substituted by two oxo groups

The "halogen" in the present invention means fluorine, chlorine, bromine and iodine.

"haloalkyl" in the context of the present invention means an alkyl group substituted with at least one halogen atom.

"haloalkoxy" in the context of the present invention means an alkoxy group substituted by at least one halogen, preferably C substituted by at least one halogen_1-6Alkoxy, more preferably C substituted by at least one halogen_1-3Alkoxy, suitably halo C_1-3Alkoxy is chloromethoxy, fluoromethoxy, dichloromethoxy, difluoromethoxy, trichloromethoxy or trifluoromethoxy; dichloroethoxy, difluoroethoxy, trichloroethoxy, trifluoroethoxy.

"Monoalkylamino" according to the invention means-NH-alkyl, preferably-NH-C_1-6Alkyl, more preferably-NH-C_1-3An alkyl group.

"dialkylamino" in the context of the present invention means-N- (alkyl), preferably-N- (C)_1-6Alkyl) (C_1-6Alkyl group), more preferably-N- (C)_1-3Alkyl) (C_1-3Alkyl groups).

"aminoalkyl" according to the invention means-alkyl-NH₂。

"Monoalkylaminoalkyl" according to the present invention refers to-alkyl-NH-alkyl.

"Dialkylaminoalkyl" in the context of the present invention refers to-alkyl-N- (alkyl).

"hydroxyalkyl" in the context of the present invention means-alkyl-OH.

"Alkylacyl" as used herein refers to-C (O) -alkyl.

"aminoacyl" according to the invention is-C (O) -NH₂。

"Monoalkylaminoacyl" according to the present invention refers to-C (O) -NH-alkyl.

"Dialkylaminoacyl" according to the invention is intended to mean-C (O) -N- (alkyl).

"Heterocycloalkyl" as used herein refers to a substituted or unsubstituted, saturated, partially saturated, cyclic alkyl group containing at least one heteroatom selected from the group consisting of N, O, S.

The "aryl" of the present invention refers to an aromatic system which may comprise a single or multiple condensed rings such as bicyclic or tricyclic aromatic rings, wherein at least a portion of the condensed rings form a conjugated aromatic system containing 5 to 50 carbon atoms, preferably about 6 to about 14 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, anthracenyl, tetrahydronaphthyl, fluorenyl, indanyl, biphenylenyl, and acenaphthenyl. "C" of the invention₆-C₁₀Aryl "refers to aromatic systems containing 6 to 10 carbon atoms.

The term "heteroaryl" as used herein refers to an aromatic group having at least one carbon atom of an aromatic monocyclic or polycyclic ring such as bicyclic or tricyclic ring replaced by a heteroatom, said heteroatom being O, S, N. Suitable heteroaryl groups include, but are not limited to, imidazolyl, benzimidazolyl, imidazopyridinyl, quinazolinyl, pyrrolyl, imidazolonyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, and the like. "C" of the invention₅-C₁₀Heteroaryl "refers to heteroaryl groups containing 5 to 10 atoms.

"C" of the invention₄-C₈"Nitrogen-containing heterocyclic group" means a substituted or unsubstituted, saturated, partially saturated, and fully unsaturated heterocyclic group containing at least one nitrogen atom and having a total number of ring atoms of 4,5, 6,7, or 8, e.g., C₄The nitrogen-containing heterocyclic group means a substituted or unsubstituted saturated, partially saturated and fully unsaturated nitrogen-containing heterocyclic group having a total number of ring atoms of 4. "C" of the invention₄-C₈Nitrogen-containing heteromonocycloalkyl "refers to substituted or unsubstituted saturated, partially saturated, and fully unsaturated, monoheterocycloalkyl having a total number of ring atoms of 4,5, 6,7, or 8 containing at least one nitrogen atom, suitable examples include, but are not limited to, substituted or unsubstituted azetidinyl, azetyl, azacyclohexyl, azepinyl, azooctyl, azaoxocyclopentyl, azaoxohexyl, azaoxoheptyl, azooctyl, azothiocyclopentyl, azothiocyclohexylA nitrogen sulfur heterocyclic heptyl group, a nitrogen sulfur heterocyclic octyl group, a diazacyclopentyl group, a diazacyclocyclohexyl group, a diazacyclocycloheptyl group or a diazacyclooctyl group; and substituted or unsubstituted nitrogen-containing heteroaryl groups, such as substituted or unsubstituted pyridyl, imidazolyl, pyrazolyl, pyrrolyl, pyridazinyl, pyrimidinyl, pyrazinyl, wherein the substituents are selected from alkyl, oxo, cycloalkyl, hydroxy, hydroxyalkyl, alkoxy, amino, monoalkylamino, dialkylamino, amido, alkylamido, arylamido, heteroarylamido, halogen, haloalkyl, haloalkoxy. Herein, said C₄-C₈The nitrogen-containing heterocyclic group forms an azacycloindole structure with the indole ring, for example, suitable azacycloindole structures include, but are not limited to

And the like.

Detailed Description

The following representative examples are intended to better illustrate the present invention and are not intended to limit the scope of the present invention.

Example 1 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (2, 3-dihydro-1H-pyrrolo [1,2-a ] -indol-9-yl) pyrimidin-2-yl) amino) phenyl) allylamide

Step a Synthesis of 1- (3-bromopropyl) -1H-indole

Adding NaH (60% content, 1.23g,30.73mmol) and DMF (10mL) in sequence into a 100mL reaction bottle, stirring at room temperature for 5min, cooling to 0-4 ℃, slowly adding 10mL of DMF solution dissolved with indole (3g,25.61mmol), after the addition is finished, heating to room temperature and reacting for 20min to obtain an indole activated solution.

Another 250ml reaction flask was charged with 1, 3-dibromopropane (15.51g,76.82mmol) and DMF (50 ml). Slowly dripping the prepared indole activating solution at 0-4 ℃, and reacting for 0.5h at room temperature after dripping. After the reaction, water (100ml) was added to quench the reaction, ethyl acetate was extracted, the ethyl acetate layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the title compound.

ESI-Ms m/z:239.1[M+H]。

Step b Synthesis of 1- (3-iodopropyl) -1H-indole

In a 250ml reaction flask, the product of step a, 1- (3-bromopropyl) -1H-indole (4.72g,19.83mmol), sodium iodide (13.39g,89.93mmol) and acetonitrile (100ml) were added in this order and refluxed overnight. After the reaction, water was added, ethyl acetate was extracted, and the ethyl acetate layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated and dried to give the title compound which was used in the next step.

ESI-Ms m/z:286.1[M+H]。

Step c Synthesis of 2, 3-dihydropyrrolo [1,2-a ] indole

1- (3-iodopropyl) -1H-indole (5.44g,19.83mmol), potassium phosphate (8.4g, 39.67mmol), tetrakistriphenylphosphine palladium (2.3g,1.98mmol) and 1, 4-dioxane (80ml) from step b were added sequentially in a 250ml three-necked flask, under argon atmosphere, and refluxed overnight. After the reaction is finished, water is added to quench the reaction, ethyl acetate is used for extraction, an ethyl acetate layer is combined, anhydrous sodium sulfate is used for drying, filtering, concentrating and column chromatography purification are carried out, and the title compound is obtained.

ESI-Ms m/z:158.1[M+H]。

Step d Synthesis of 9- (2-Chloropyrimidin-4-yl) -2, 3-dihydropyrrolo [1,2-a ] indole

In a 100ml reaction flask, aluminum trichloride (2.18g,16.35mmol), ethylene glycol dimethyl ether (50ml), 2, 4-dichloropyrimidine (2.44g,16.35mmol) and 2, 3-dihydropyrrolo [1,2-a ] indole (2.57g,16.35mmol) obtained in step c were added in this order, and the reaction was refluxed for 2 hours. After the reaction is finished, the reaction solution is cooled to room temperature, filtered, washed by filter cake water and dried to obtain the subject.

ESI-Ms m/z:270.4[M+H]。

Step e N Synthesis of- (4-fluoro-2-methoxy-5-nitrophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine

And d, adding the 9- (2-chloropyrimidin-4-yl) -2, 3-dihydropyrrolo [1,2-a ] indole (2.69g, 10mmol), 4-fluoro-2-methoxy-5-nitroaniline (1.86g, 10mmol) and p-toluenesulfonic acid (1.71g, 10mmol) obtained in the step d into a 250mL reaction flask, adding 50mL of sec-butyl alcohol to dissolve, reacting at 110 ℃ for 2h, cooling to room temperature after the reaction is finished, filtering, washing with sec-butyl alcohol, and drying to obtain the title compound.

ESI-Ms m/z:420.4[M+H]。

Step f N Synthesis of- (4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitrophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine

And (3) sequentially adding the N- (4-fluoro-2-methoxy-5-nitrophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine (2.5g, 6mmol), N, N, N' -trimethylethylenediamine (0.61g, 6mmol), diisopropylethylamine (2.3g, 18mmol) and 10ml of dioxane into a 50ml reaction flask, refluxing at 110 ℃ for 3h, concentrating after the reaction is finished, and purifying by column chromatography to obtain the title compound.

ESI-Ms m/z:502.4[M+H]。

Step g N Synthesis of- (4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-aminophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine

The product obtained in step f, N- (4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-nitrophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine (2.5g, 5mmol), 10% Pd-C (20mg) and 30ml methanol were added sequentially in a 50ml reaction flask and reduced with hydrogen for 1h at 1 atm, after the reaction was complete, filtered and concentrated to give the title compound, which was used directly in the next step.

ESI-Ms m/z:472.6[M+H]。

Step H N Synthesis of- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((5-methyl-4- (2, 3-dihydro-1H-pyrrolo [1,2-a ] -indol-9-yl) pyrimidin-2-yl) amino) phenyl) allylamide

In a 150ml single-neck flask, the product obtained in step g, N- (4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxy-5-aminophenyl) -4- (2, 3-dihydropyrrolo [1,2-a ] indol-9-yl) -pyrimidin-2-amine (2g, 4.2mmol), diisopropylethylamine (0.53g, 4.2mmol) and 20ml of anhydrous dichloromethane were added, after dissolution, 5ml of a dichloromethane solution in which allyl chloride (0.37g, 4.2mmol) was dissolved was slowly added, reaction was carried out for 10min, after completion of the reaction, concentration was carried out, and column chromatography was carried out to obtain the title compound.

¹H NMR(300MHz,DMSO-d₆):10.17(s,1H),8.69(s,1H),8.28-8.25(d,2H),8.02(s,1H),7.37-7.35(d,1H),7.15-6.97(m,4H),6.44-6.35(m,1H),6.21-6.15(m,1H)，5.74-5.70(d,1H),4.1-4.10(t,2H),3.81(s,3H),3.35(s,1H),3.30-3.25(t,1H),2.90(s,2H),2.73(s,3H),2.60-2.56(m,2H),2.33(s,2H),2.22(s,6H)。

ESI-Ms m/z:526.4[M+H]。

Example 2 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

Step a Synthesis of 1- (4-bromobutyl) -1H-indole

Sequentially adding NaH (60% content, 1.23g,30.73mmol) and DMF (10mL) into a 100mL reaction bottle, stirring at room temperature for 5min, cooling to 0-4 ℃, slowly adding 10mL of DMF solution dissolved with indole (3g,25.61mmol), heating to room temperature after addition, and reacting for 20min to obtain an indole activation solution.

Another 250mL reaction flask was charged with 1, 4-dibromobutane (16.59g,76.82mmol) and DMF (50 mL). Slowly dripping the prepared indole activating solution at 0-4 ℃, and reacting for 0.5h at room temperature after dripping. After the reaction, water (100mL) is added for quenching, ethyl acetate is used for extraction, ethyl acetate layers are combined, anhydrous sodium sulfate is dried, and the mixture is filtered, concentrated and purified by column chromatography to obtain the title compound.

ESI-Ms m/z:252.1[M+H]。

Step b Synthesis of 1- (4-iodobutyl) -1H-indole

To a 250mL reaction flask, 1- (4-bromobutyl) -1H-indole (5g,19.83mmol), sodium iodide (13.39g,89.93mmol), and acetonitrile (100mL) from step a were added in that order and refluxed overnight. After the reaction, water was added, ethyl acetate was extracted, and the ethyl acetate layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated and dried to give the title compound which was used in the next step.

ESI-Ms m/z:300.0[M+H]。

Step c Synthesis of 6,7,8, 9-tetrahydropyrido [1,2-a ] indole

1- (4-iodobutyl) -1H-indole (5.93g, 19.83mmol), potassium phosphate (8.4g, 39.67mmol), tetrakistriphenylphosphine palladium (2.3g,1.98mmol), 1, 4-dioxane (80mL), which was obtained in step b, was added in succession to a 250mL three-necked flask, under argon protection, and refluxed overnight. After the reaction is finished, water is added to quench the reaction, ethyl acetate is used for extraction, an ethyl acetate layer is combined, anhydrous sodium sulfate is used for drying, filtering, concentrating and column chromatography purification are carried out, and the title compound is obtained.

ESI-Ms m/z:172.1[M+H]。

Step d Synthesis of 10- (2-Chloropyrimidin-4-yl) -6,7,8, 9-tetrahydropyrido [1,2-a ] indole

In a 100mL reaction flask, aluminum trichloride (2.18g,16.35mmol), ethylene glycol dimethyl ether (50mL), 2, 4-dichloropyrimidine (2.44g,16.35mmol) and 6,7,8, 9-tetrahydropyrido [1,2-a ] indole (2.8g,16.35mmol) obtained in step c were sequentially added, and the reaction was refluxed for 2 hours. After the reaction is finished, the reaction solution is cooled to room temperature, filtered, washed by filter cake water and dried to obtain the subject.

ESI-Ms m/z:284.1[M+H]。

Step e N Synthesis of- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared according to the methods of example 1, steps e, f, g and h using 10- (2-chloropyrimidin-4-yl) -6,7,8, 9-tetrahydropyrido [1,2-a ] indole, 4-fluoro-2-methoxy-5-nitroaniline, N' -trimethylethylenediamine, allyl chloride, which was the product of step d, as the starting material.

¹H NMR(300MHz,DMSO-d₆)10.20(s,1H),8.65(s,1H),8.34(d,1H),8.11(s,1H),8.06(d,1H),7.43(d,1H),7.19-7.03(m,3H),6.98(s,1H),6.57-6.41(m,1H),6.28-6.15(m,1H),5.82-5.71(m,1H),4.09(t,2H),3.84(s,3H),3.18(t,2H),3.06-2.92(m,2H),2.66(s,3H),2.47-2.40(m,2H),2.27(s,6H),2.08-1.96(m,2H),1.87-1.74(m,2H)。

ESI-Ms m/z:540.3[M+H]。

Example 3 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((5-methyl-4- (6,7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

Step a Synthesis of 10- (2-chloro-5-methylpyrimidin-4-yl) -6,7,8, 9-tetrahydropyrido [1,2-a ] indole

The title compound was prepared as in example 2 step d starting from 6,7,8, 9-tetrahydropyrido [1,2-a ] indole and 5-methyl-2, 4-dichloropyrimidine, obtained in example 2 step c.

Step b N Synthesis of- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((5-methyl-4- (6,7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared according to the methods of example 1, steps e, f, g and h starting from 10- (2-chloro-5-methylpyrimidin-4-yl) -6,7,8, 9-tetrahydropyrido [1,2-a ] indole, 4-fluoro-2-methoxy-5-nitroaniline, N' -trimethylethylenediamine and allyl chloride, which was obtained in step a.

¹H NMR(300MHz,DMSO-d₆)10.02(s,1H),8.84(s,1H),8.33(s,1H),7.77(s,1H),7.42(d,1H),7.26(d,1H),7.10-7.13(m,1H),7.04-7.07(m,1H),6.95(s,1H),6.34-6.39(m,1H),6.19-6.22(m,1H),5.73(d,1H),4.11(t,2H),3.83(s,3H),2.99(s,2H),2.83(t,2H),2.67(s,3H),2.25(t,2H),2.17(s,6H),2.07(s,3H),2.04(m,2H),1.79(m,2H)。

ESI-Ms m/z:554.3[M+H]。

Example 4 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] -5-fluoroindol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared as in example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 5-fluoroindole, 1, 4-dibromobutane, N' -trimethylethylenediamine, and allyl chloride.

¹H NMR(300MHz,DMSO-d₆):9.62(s,1H),9.42(s,1H),8.3-8.27(d,2H),7.87(s,1H),7.49(d,1H),7.14(d,1H),7.06(s,2H),6.71-6.63(m,1H),6.22-6.16(m,1H)5.73(d,1H),4.13(s,2H),3.84(s,3H),3.32(s,4H),3.22-3.20(t,2H),2.83(s,6H),2.64(s,3H),2.02(s,2H),1.85(s,2H)。

ESI-Ms m/z:558.4[M+H]。

Example 5 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- ((2, 3-dihydro-1H-pyrrolo [1,2-a ] -5-fluoroindol-9-yl) -5-chloro-pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared as in example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2,4, 5-trichloropyrimidine, 5-fluoroindole, 1, 3-dibromopropane, N' -trimethylethylenediamine, allyl chloride.

1H NMR(300MHz,DMSO-d6)：9.99(s,1H),8.48(s,1H),8.42(s,2H),7.41-7.34(m,2H),6.98-6.91(m,2H),6.44–6.39(m,1H),6.17(m,1H),5.61(d,1H),4.14(t,2H),3.82(s,3H),3.16–3.11(m,2H),2.91-2.87(m,2H),2.68(s,3H),2.57–2.54(m,2H),2.39–2.35(m,2H),2.23(s,6H).

ESI-Ms m/z:578.2[M+H]。

Example 6 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (2, 3-dihydro-1H-pyrrolo [1,2-a ] -5-fluoroindol-9-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared as in example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 5-fluoroindole, 1, 3-dibromopropane, N' -trimethylethylenediamine, and allyl chloride.

1H NMR(300MHz,DMSO-d6)10.07(s,1H),8.65(s,1H),8.25(d,1H),8.05–8.02(m,1H),7.36-7.34(m,2H),7.01(s,1H),6.90–6.97(m,2H),6.36-6.34(m,1H),6.17-6.15(m,1H),5.69-5.67(d,1H),4.13(t,2H),3.79(s,3H),2.89–2.88(t,2H),2.73(s,3H),2.62–2.60(t,2H),2.51–2.50(t,2H),2.32-2.30(t,2H),2.21(s,6H).

ESI-Ms m/z:544.4[M+H]。

Example 7 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (3, 4-dihydro-1H- [1,4] oxazino [4,3-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

Step a Synthesis of 2-hydroxymethylindole

Lithium aluminum hydride (902.63mg,23.78mmol) and THF (30ml) were added sequentially to a 100ml reaction flask, stirred at room temperature for 5min, then cooled in an ice bath, ethyl indole-2-carboxylate (3g,15.86mmol) was slowly added, and after the addition, the temperature was raised to room temperature for reaction for 2 h. After the reaction was completed, the system was cooled to 0 ℃ and then 20ml of THF and 1.7ml of 20% KOH were added thereto, stirred for 10min and filtered, the filter cake was washed with 20ml of THF, the filtrate was washed once with saturated NaCl (10ml), dried over anhydrous sodium sulfate, filtered, concentrated and dried to obtain the title compound 2.3g, which was used directly in the next step.

ESI-Ms m/z:148.1[M+H]。

Step b Synthesis of 3, 4-dihydro-1H- [1,4] oxazino [4,3-a ] indole

In a 500ml reaction flask, the 2-hydroxymethylindole (300mg,2.04mmol), KOH (285.91mg,5.10mmol) and CH obtained in step a were added in this order₂Cl₂(160ml), cooled to about 0 ℃ in an ice bath, protected by nitrogen, and stirred for 10 min. 40mL of CH containing Diphenyl (vinyl) sulfonium trifluorinate (888.86mg,2.45mmol) dissolved therein₂Cl₂And dropwise adding the solution into the reaction system, and heating to room temperature for reaction after the addition is finished. When TLC monitored disappearance of starting material, after reaction was complete, water was added to quench the reaction, ethyl acetate was extracted, the ethyl acetate layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give 300mg of the title compound.

ESI-Ms m/z:174.1[M+H]。

Step c N Synthesis of- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (3, 4-dihydro-1H- [1,4] oxazino [4,3-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared according to the procedure for example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 3, 4-dihydro-1H- [1,4] oxazino [4,3-a ] indole, N' -trimethylethylenediamine, and allyl chloride, as obtained in step b.

1H NMR(300MHz,DMSO-d6)10.04(s,1H),8.61(s,1H),8.35(d,1H),8.06(d,1H),8.03(s,1H),7.50-7.49(d,1H),7.24–7.19(m,2H),7.12-7.11(d,1H),7.00(s,1H),6.45(s,1H),6.22–6.19(d,1H),5.73-5.71(d,1H),5.07(s,2H),4.16–4.14(t,2H),4.06–4.04(t,2H),3.81(s,3H),2.96(s,2H),2.70(s,3H),2.30(s,6H),2.35(m,2H).

ESI-Ms m/z:542.2[M+H]。

Example 8 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] -7-chloroindol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared as in example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 7-chloroindole, 1, 4-dibromobutane, N' -trimethylethylenediamine, and allyl chloride.

1H NMR(300MHz,DMSO-d6)：10.04(s,1H),8.69(s,1H),8.38–8.36(d,1H),8.26(s,1H),8.00(s,2H),7.14–7.12(m,1H),7.03–6.98(m,2H),6.43–6.35(m,1H),6.17–6.16(d,1H),5.71–5.70(d,1H),4.66–4.62(m,2H),3.34–3.21(m,3H),2.90–2.86(m,3H),2.70(s,4H),2.33–2.31(m,2H),2.20(s,6H),2.20–2.19(m,2H),1.78–1.76(m,2H),

ESI-Ms m/z:575.2[M+H]。

Example 9 Synthesis of N- (5- ((4- (1, 1-dimethyl-2, 3-dihydro-1H-pyrrolo [1,2-a ] indol-9-yl) pyrimidin-2-yl) amino) -2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxyphenyl) acrylamide

The title compound was prepared according to the procedure for example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 1, 3-dibromo-3-methylbutane, indole, N' -trimethylethylenediamine, and allyl chloride.

1H NMR(300MHz,DMSO-d6)10.00(s,1H),8.58(s,1H),8.36–8.58(m,1H),7.91–7.96(m,2H),7.35–7.40(m,1H),7.09–7.40(m,4H),6.36–6.45(m,1H),6.18–6.23(m,1H),5.71–5.74(m,1H)4.12–4.19(m,2H),3.80(s,3H),2.93(s,2H),2.69(s,3H),2.37–2.44(m,2H),2.28(s,6H),1.56(s,2H),1.37(s,6H).

ESI-Ms m/z:554.3[M+H]。

Example 10 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -5-chloro-pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared in accordance with the procedure for example 1 starting from 6,7,8, 9-tetrahydropyrido [1,2-a ] indole, 4-fluoro-2-methoxy-5-nitroaniline, 2,4, 5-trichloropyrimidine, N' -trimethylethylenediamine and allyl chloride, which was obtained in step c of example 2.

1H NMR(300MHz,DMSO-d6)：9.63(s,1H),9.47(s,1H),8.3–8.30(d,2H),7.85(s,1H),7.5(d,1H),7.12(d,1H),7.04(s,2H),6.72–6.61(m,1H),6.23–6.15(m,1H)，5.7,5(d,1H),4.12(s,2H),3.86(s,3H),3.34(s,4H),3.20–3.17(t,2H),2.86(s,6H),2.66(s,3H),2.01(s,2H),1.86(s,2H).

ESI-Ms m/z:575.2[M+H]。

Example 11 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6,7,8, 9-tetrahydropyrido [1,2-a ] -5-methoxy-indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

The title compound was prepared as in example 1 starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 5-methoxyindole, 1, 4-dibromobutane, N' -trimethylethylenediamine, and allyl chloride.

1H NMR(300MHz,DMSO-d6)：10.02(s,1H),9.53(s,1H),8.40–8.45(d,2H),7.92(s,1H),7.52(d,1H),7.18(d,1H),7.10(s,2H),6.76–6.70(m,1H),6.28–6.20(m,1H)5.72(d,1H),4.12(s,2H),3.85(s,3H),3.72(s,3H),3.32(s,4H),3.21–3.19(t,2H),2.83(s,6H),2.65(s,3H),2.02(s,2H),1.86(s,2H).

ESI-Ms m/z:570.4[M+H]。

Example 12 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino-4-methoxy-5- ((4- (1,1a,2,8 b-tetrahydrocyclopropyl [3,4] pyrrolo [1,2-a ] indol-8-yl) pyrimidin-2-yl) amino) phenyl) acrylamide

Step a 1-allyl-1H-indole-2-carbaldehyde synthesis:

in a 100mL double-necked flask, 25mL of anhydrous DMF was added, cooled to 0-5 deg.C, and 662mg of NaH was slowly added, stirred for 10 min. Slowly dripping 10mL of anhydrous DMF solution dissolved with 2g of 1H-indole-2-formaldehyde into the reaction solution to generate bubbles, changing the color of the solution into dark black, continuously stirring for 15min, then slowly adding 5mL of anhydrous DMF solution dissolved with 1.82g of 3-bromopropylene, reacting for 3H at room temperature, adding 50mL of water, extracting with 3X60mL ethyl acetate, concentrating, drying, and carrying out column chromatography to obtain 2.0g of the title compound.

ESI-Ms m/z:186.0[M+H]

Step b: synthesis of N- ((1-propenyl-1H-indol-2-yl) methylene) -4-methylbenzenesulfonylhydrazide:

in a 100mL single-neck flask, 1-allyl-1H-indole-2-carbaldehyde obtained in step a (1.85g,10mmol), p-toluenesulfonylhydrazide (1.86g, 10mmol) and 50mL of methanol were sequentially added, and the reaction was refluxed at 85 ℃ in an oil bath for 2 hours. Concentration gave 3.6g of the title crude product, which was used directly in the next step.

Step c: synthesis of 1,1a,2,8 b-tetrahydrocyclopropyl [3,4] pyrrolo [1,2-a ] indole:

in a 250mL three-necked flask, N- ((1-propenyl-1H-indol-2-yl) methylienyl) -4-methylbenzenesulfonyl hydrazide obtained in the step b (2.2g,6.23mmol), potassium carbonate (1.29g, 9.35mmol) and 60mL of dioxane were added, and the reaction was refluxed at 100 ℃ in an oil bath for 12 hours. Filtration, concentration, addition of 50ml water, 3X 40ml ethanol extraction, 30ml water washing, concentration, column chromatography to obtain 1.0g title product, directly used in the next step.

Step d: synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino-4-methoxy-5- ((4(1,1a,2,8 b-tetrahydrocyclopropyl [3,4] pyrrolo [1,2-a ] indol-8-yl) pyrimidin-2-yl) amino) phenyl) acrylamide

The title compound was synthesized according to the procedure for the synthesis of example 1, starting from 4-fluoro-2-methoxy-5-nitroaniline, 2, 4-dichloropyrimidine, 1a,2,8 b-tetrahydrocyclopropyl [3,4] pyrrolo [1,2-a ] indole, N' -trimethylethylenediamine and allyl chloride from step c.

¹H-NMR(300MHz,DMSO-d₆):10.03(s,1H),8.70(s,1H),8.32-8.33(d,1H),8.24-8.26(d,1H),8.01(s,1H),7.21-7.28(m,2H),7.10-7.13(m,1H),7.03-7.06(m,2H),6.41-6.47(m,1H),6.20-6.23(d,1H),5.74-5.76(d,1H),4.19-4.28(m,2H),3.86(s,3H),2.97(s,2H),2.95(s,2H),2.74(s,3H),2.29-2.5(m,8H),1.44-1.45(m,1H),0.70-0.71(m,1H)。

ESI-Ms m/z:537.9[M+H]

Example 13 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6-oxo-6, 7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:554.4[M+H]

Example 14 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (6-fluoro-6, 7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:558.3[M+H]

Example 15 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (9-fluoro-6, 7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:558.5[M+H]

Example 16 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (9, 9-difluoro-6, 7,8, 9-tetrahydropyrido [1,2-a ] indol-10-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:575.3[M+H]

Example 17 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1, 2-dihydroazetidino [1,2-a ] indol-8-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:511.6[M+H]

Example 18 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1-fluoro-2, 3-dihydro-1H-pyrrolo [1,2-a ] -indol-9-yl) pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:543.8[M+H]

Example 19 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (2-oxo-1, 2-dihydroazetidino [1,2-a ] indol-8-yl) -pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:525.4[M+H]

EXAMPLE 20 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1, 1-dioxo-3, 4-dihydro-2H- [1,2] thiazin [2,3-a ] indol-5-yl) pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:590.4[M+H]

Example 21 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (2, 3-dimethyl-1, 2,3, 4-tetrahydropyrazino [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:569.5[M+H]

Example 22 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (1-oxo-1, 2,3, 4-tetrahydropyrazin [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:555.7[M+H]

Example 23 Synthesis of N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -4-methoxy-5- ((4- (2-methyl-1, 2,3, 4-tetrahydropyrazin [1,2-a ] indol-10-yl) pyrimidin-2-yl) amino) phenyl) allylamide

ESI-Ms m/z:554.8[M+H]

Experimental example 1 in vitro kinase Activity evaluation

1 materials of the experiment

1.1 enzymes

EGFR^WTKinases available from Carna corporation;

EGFR^T790M/L858Rkinase, available from Invitrogen.

1.2 reagents

Adenosine Triphosphate (ATP), purchased from Sigma;

peptide (Peptide FAM-P22) available from GL Biochem;

ethylenediaminetetraacetic acid (EDTA), available from Sigma.

1.3 instruments

Caliper EZ reader microfluidic chip instrument, available from Caliper Life Sciences, Inc.

2 method of experiment

2.1 preparation of 1 Xkinase base buffer and stop buffer

1 × kinase base buffer (for EGFR)^WT)：50mM HEPES，pH7.5，0.0015％Brij-35，10mMMgCl₂，10mM MnCl₂，2mM DTT；

1 × kinase base buffer (for EGFR)^T790M/L858R)：50mM HEPES，pH7.5，0.0015％Brij-35，10mM MgCl₂，2mM DTT；

Stop buffer: 100mM HEPES, pH7.5, 0.0015% Brij-35, 0.2% Coating Reagent #3, 50mM EDTA.

2.2 preparation of Compounds

The compounds of the invention were dissolved in 100% DMSO to 10mM each, diluted to 50. mu.M in complete medium, then diluted to 5. mu.M in complete medium containing 0.1% DMSO, and then sequentially diluted 3-fold for 10 concentrations (for EGFR)^WT)；

The compounds of the invention were dissolved separately in 100% DMSO10mM, diluted to 50. mu.M in complete medium, then diluted to 1. mu.M in complete medium containing 0.1% DMSO, and then diluted 3-fold in sequence for a total of 10 concentrations (for EGFR)^T790M ^/L858R)；

Add 100. mu.l 100% DMSO to empty wells for preparing kinase no compound control and kinase no compound control;

the 96-well plate used above was labeled as the source plate.

2.3 preparation of the intermediate plate

Transfer 10. mu.l of the solution from the source plate to a new 96-well plate as an intermediate plate, add 90. mu.l of 1 Xkinase buffer to each well of the intermediate plate, and mix by shaking for 10 min.

2.4 preparation of the test plate

From a 96-well intermediate plate, 5. mu.l of the solution was transferred per well to a 384-well plate.

2.5 kinase reaction

2.5.1. Preparation of 2.5 × kinase solution EGFR^WTKinase and EGFR^T790M/L858RAdding the kinase stock solution into 1 × basic buffer solution respectively to prepare 2.5 × kinase solution;

2.5.2. preparation of a 2.5 Xpeptide solution: adding FAM labeled peptide and ATP into 1 × basic buffer solution to prepare 2.5 × peptide solution;

2.5.3. transfer 10. mu.l of 2.5 Xkinase solution to 384 well assay plates and incubate for 10min at room temperature;

2.5.4. transfer 10. mu.l of 2.5 Xpeptide solution to 384 well plates, incubate at 28 ℃ for a period of time, and stop the reaction by adding 25. mu.l of stop buffer.

A kinase-free no compound control group (containing DMSO, 1 Xbase buffer and 2.5 Xpeptide solution) and a kinase-free no compound control group (containing DMSO, 2.5 Xkinase solution and 2.5 Xpeptide solution) were set at the same time.

Reading and fitting a curve of the Caliper instrument, and calculating the inhibition rate

Reading data on a Caliper instrument, obtaining conversion data from a Caliper program, and calculating the inhibition rate according to the following formula:

inhibition rate [% ], [% ] represents [% ], [% ] represents "[% ], [% ] represents" [% ].

2.5.6. Computing IC Using Graphpad 5.0 data processing software₅₀The value is obtained. The results are shown in Table 1.

TABLE 1

From the results of the experiments, it is clear that the compounds of the present invention are active against mutant EGFR kinases, such as EGFR^L858R/T790MThe kinase has good inhibitory activity, IC₅₀The value is less than 1nM, the influence on wild type EGFR kinase is small, and the selectivity is good.

Experimental example 2 evaluation of in vitro cell Activity

2. Experimental Material

1.1 cells and Positive control

Experimental cell lines NCI-H1975(EGFR double mutant cells with L858R and T790M mutations) and A431(EGFR wild-type cells) were purchased from ATCC.

The commercial highly selective irreversible EGFR mutant inhibitor AZD9291, produced by astrazeneca, was used as a positive control, and the synthetic method was as described in PCT/GB2012/051783 (patent No. WO2013/014448) specification example 28, and confirmed by hydrogen spectroscopy and mass spectroscopy.

1.2 reagents

Cell Titer-Glo luminescence Cell viability assay, available from Promega corporation;

RPMI1640medium, available from Invitrogen;

DMEM medium, available from Invitrogen corporation;

fetal bovine serum, purchased from Invitrogen;

DMSO, available from Sigma company;

NCI-H1975 cells were cultured in RPMI1640medium containing 10% inactivated fetal bovine serum (GIBCO) containing 100IU/mL of penicillin and 100. mu.g/mL of streptomycin;

a431 cells were cultured in DMEM medium containing 10% inactivated fetal bovine serum (GIBCO) containing 100IU/mL of penicillin and 100. mu.g/mL of streptomycin.

2 method of experiment

2.1 Experimental Process (CTG assay)

NCI-H1975 cells and A431 cells in logarithmic growth phase were digested, blown into single cell suspension, seeded in 96-well culture plates with 100. mu.L of medium per well, and 3 pieces of 96-well plates per cell line, wherein NCI-H1975 cells were seeded at 3X10 per well³Individual cells, a431 cells were seeded at 4X 10 per well³And (4) cells. Inoculating NCI-H1975 cells and A431 cells in 5% CO₂After the cells are attached to the surface, the test compound (the highest test concentration of the compound on NCI-H1975 cells is 4 mu M, 3 times of dilution, and 9 concentrations in total; the highest test concentration on A431 cells is 10 mu M, 3 times of dilution, and 9 concentrations in total) is added, and the cells are cultured in the incubator for another 72 hours. Both a blank control (medium only, no cells and DMSO solution) and a DMSO control (medium with cells and 0.5% DMSO solution) were set. Add 100. mu.L of CTG solution, shake for 2min in the dark, incubate for 10 min. 2.2 reading, calculating IC₅₀Value of

Placing the culture plate in

Reading the plate by a multi-mode microplate detector, recording the luminescence reading result, and calculating the inhibition rate according to the following formula:

inhibitor (%) - (1- (RLU)_com-RLU_blank)/(RLU_DMSO–RLU_blank))×100％，

Wherein RLU_comDenotes the absorbance, RLU, of the test Compound group_blankAbsorbance values, RLU, of blank control group_DMSOThe absorbance of the DMSO control was expressed,

the drug effect inhibition rate curve is drawn by using XLFit curve fitting software and the IC is calculated₅₀The values and results are shown in Table 2.

TABLE 2

In vitro cell experiment results show that the compound of the invention has better inhibition effect on double mutant cells (NCI-H1975), has small inhibition effect on EGFR wild type cells (A431), and has selectivity similar to or obviously superior to AZD 9291. One of the major side effects of currently marketed EGFR inhibitors is known to be skin rash, diarrhea, etc., which are associated with poor selectivity due to the inhibition of wild-type EGFR. Therefore, the compound of the invention is expected to be a medicine which has specific curative effect on resisting tumors with drug resistance caused by EGFR mutation and has small side effect.

Experimental example 3 blood sugar Effect test

1 materials of the experiment

1.1 Compounds

Dissolving the compound with 25mM citric acid-sodium citrate buffer solution (pH4.5), preparing into 1.25mg/mL clear solution, and setting as test compound group; using 25mM citric acid-sodium citrate buffer solution (pH4.5) as solvent control group;

1.2 animals

Female BALB/C mice, each 5 in each group, weighing 18-22g, were provided by Shanghai Sphere-BiKai laboratory animals Co. The test mice are given an environmental adaptation period of 2-4 days before the experiment, and are fasted for 8-12h before the administration.

2 method of experiment

2.1 mice fasted but had free access to water for 12 hours;

2.2 taking 5 mice from step 2.1, intragastric administration (I.G.) 25mg/kg of the compound of the present invention;

2.3 blood is collected at eye orbit 0min, 15min, 30min, 1h and 2h after gastric lavage, the blood sugar value A of the animal is detected by using a Roche excellence type glucometer, and the AUC is [ (A) according to the formula_15min+A₀)×7.5]+[(A_30min+A_15min)×7.5]+[(A_60min+A_30min)×15]+[(A_120min+A_60min)×30]Calculating AUC value, and increasing AUC%_{Compound (I)}－AUC_Solvent)/AUC_Solvent]× 100 AUC increase calculated by 100%Length of which is "A₀"represents the blood sugar level 0min after intragastric administration," A_15min"represents the blood sugar value after intragastric administration for 15min," A_30min"represents the blood sugar value after 30min of intragastric administration," A_60min"represents the blood sugar value 60min after intragastric administration," A_120min"represents blood sugar value 120min after intragastric administration," AUC_{Compound (I)}"AUC value representing test Compound," AUC_Solvent"represents the AUC value of the vehicle control group, and the results are shown in Table 3.

TABLE 3

EGFR inhibitors are known to cause side effects such as hyperglycemia and impaired insulin signaling in vivo. According to the experimental reports, AZD9291 and the Rociletinib (CO1686) of Clovis Oncology company have some side effect of blood sugar elevation (Management of hyperglycemic from Epidermal Growth Factor Receptor (EGFR) Tyrosine Kinase Inhibitors (TKIs) targeting T790M-dimensional Cancer, Jeryl Villadolid, et al. transformation Cancer Research (2015),4(5),576 583). The above experimental results show that the compound of the invention has little influence on blood sugar, and is expected to overcome the hyperglycemia side effect of the existing EGFR inhibitor, thereby being beneficial to reducing adverse reactions and improving the compliance of patients.

Experimental example 4 drug metabolism experiment

1 materials of the experiment

1.1 Compounds

The compounds of the present invention are formulated for both oral and parenteral administration. Wherein, the oral medicine is dissolved in 25mM citric acid-sodium citrate buffer solution (pH4.5) to prepare 1.25mg/mL clear solution; the tail vein injection drug formula is characterized in that the volume ratio of citric acid-sodium citrate buffer solution (pH4.5) to normal saline is 1: 1 to make a 0.2mg/mL solution.

1.2 animals

Female BALB/C mice, each 5 in each group, weighing 18-22g, were provided by Shanghai Sphere-BiKai laboratory animals Co. The test mice are given an environmental adaptation period of 2-4 days before the experiment, are fasted for 8-12h before the administration, are fed with water after the administration for 2h, and are fed with food after 4 h.

1.3 reagents

Methanol (chromatographically pure): manufactured by Spectrum corporation;

acetonitrile (chromatographically pure): manufactured by Spectrum corporation;

the other reagents were all commercially available analytical grade.

1.4 instruments

API 4500 model triple quadrupole LC MS, available from AB corporation, USA, equipped with electrospray ionization source (ESI), LC-30AD dual pump; SIL-30AC autosampler; a CTO-30AC column incubator; DGU-20A3R deaerator; an Analyst QS a01.01 chromatography workstation; Milli-Q ultra pure water devices (Millipore Inc); a Qilinbeier Vortex-5 oscillator; HITACHICF16R XII desk high speed refrigerated centrifuge.

2 method of experiment

2.1 after the mice are fasted but can drink water freely for 12 hours, blank plasma at 0 moment is adopted;

2.2 taking 5 mice from step 2.1, intragastric administration (I.G.) to administer 25mg/kg of the compound of the present invention;

5 mice in step 2.1 were taken and 2mg/kg of compound of the present invention was administered to the tail vein (i.v.);

2.3 after the gavage, 5min, 15min, 30min, 1h, 2h, 4h, 10h and 24h, continuously taking blood from the fundus venous plexus, placing the blood in an EP tube distributed with heparin, centrifuging at 8000rpm/min for 5min, taking upper layer plasma, freezing and storing at-20 ℃, and waiting for LC-MS/MS analysis;

continuously taking blood from the fundus venous plexus 5min, 15min, 30min, 1h, 2h, 4h, 10h and 24h after tail vein injection administration, placing the blood in an EP tube distributed with heparin, centrifuging at 8000rpm/min for 5min, taking upper layer plasma, freezing and storing at-20 ℃, and waiting for LC-MS/MS analysis;

2.4 according to the blood concentration-time data obtained in step 2.3, pharmacokinetic parameters were calculated using WinNonlin software, see table 4.

TABLE 4

The compound of the invention has better pharmacokinetic data and better clinical application prospect.

Although the present invention has been described in detail above, those skilled in the art will appreciate that various modifications and changes can be made to the present invention without departing from the spirit and scope of the invention. The scope of the invention is not to be limited by the above detailed description but is only limited by the claims.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof,

wherein:

ring A is

n is selected from 1,2,3,4, 5 and 6, m is selected from 1,2,3 and 4, R_aSelected from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl and di-C_1-6Alkylaminoacyl, when m is selected from 2,3 and 4, two R on two adjacent carbon atoms_aMay form C together with the carbon atom to which they are attached_3-8Cycloalkyl, or two R on the same carbon atom_aMay form C together with the carbon atoms to which they are jointly attached_3-8A cycloalkyl group; or

Ring A is

m is selected from 1,2,3 and 4, p is selected from 0, 1 and 2, q is selected from 0, 1 and 2, R_aSelected from hydrogen, C_1-6Alkyl radical, C_3-8Cycloalkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl and di-C_1-6Alkylaminoacyl, when m is selected from 2,3 and 4, two R on two adjacent atoms_aMay form C together with the atoms to which they are attached_3-8Cycloalkyl, or two R on the same atom_aMay form C together with the atoms to which they are commonly attached_3-8A cycloalkyl group;

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N;

R₁、R_2a、R_2b、R_3a、R_3band R_3cEach independently selected from hydrogen, halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino radical, amino radical C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl, amino acyl, mono C_1-6Alkylaminoacyl and di-C_1-6An alkylaminoacyl group;

R₄、R₅and R₆Each independently selected from hydrogen, halogen, cyano, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl radical, said C_1-6Alkyl radical, C_1-6Alkoxy and C_3-8Cycloalkyl optionally substituted by one or more halogens, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino, hydroxy, amino C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl and hydroxy C_1-6Alkyl substitution;

R₇selected from hydrogen, C_1-6Alkyl radical, C_3-6Cycloalkyl and halo C_1-6An alkyl group; and

R₈selected from hydrogen, C_1-6Alkyl and C_3-6Cycloalkyl radical, said C_1-6Alkyl and C_3-6Cycloalkyl optionally substituted by one or more halogens, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6An alkylamino group or a hydroxyl group, or a substituted group,

wherein the compound is not:

2. a compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein:

X₁、X₂、X₃and X₄Each independently selected from C (R)₁) And N; and

R₇selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and trifluoromethyl; and

R₈selected from hydrogen, C_1-3Alkyl and C_3-6Cycloalkyl radical, said C_1-3Alkyl and C_3-6Cycloalkyl is optionally substituted with one or more halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, isopropoxy, amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, dipropylamino, methylethylamino, methylpropylamino, ethylpropylamino, or hydroxy; and

R₄、R₅and R₆Each independently selected from hydrogen, halogen, cyano, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-8Cycloalkyl radical, said C_1-6Alkyl radical, C_1-6Alkoxy and C_3-8Cycloalkyl optionally substituted by one or more halogens, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, amino, mono C_1-6Alkylamino, di-C_1-6Alkylamino, hydroxy, amino C_1-6Alkyl, mono C_1-6Alkylamino radical C_1-6Alkyl, di-C_1-6Alkylamino radical C_1-6Alkyl and hydroxy C_1-6Alkyl substitution.

3. A compound, or a pharmaceutically acceptable salt thereof, which is the following compound:

4. a pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or a composition according to claim 4 in the manufacture of a medicament for the treatment and/or prophylaxis of tumours.

6. Use according to claim 5, wherein the tumour is a drug resistant tumour.

7. Use according to claim 6, wherein the tumour is a tumour resistant to EGFR inhibitors.

8. A compound of formula II or a pharmaceutically acceptable salt thereof,

wherein the ring A, X₁、X₂、X₃、X₄、R_2a、R_2b、R_3a、R_3b、R_3c、R₇And R₈Has the definition of any one of claims 1 to 3, and

the compound is not: