CN108721298A

CN108721298A - As the pyrimido heterocyclic compound of bruton's tyrosine kinase inhibitor and its application

Info

Publication number: CN108721298A
Application number: CN201710258052.2A
Authority: CN
Inventors: 李洪林; 丁健; 徐玉芳; 谢华; 赵振江; 陈海洋; 刁妍妍
Original assignee: East China University of Science and Technology; Shanghai Institute of Materia Medica of CAS
Current assignee: East China University of Science and Technology; Shanghai Institute of Materia Medica of CAS
Priority date: 2017-04-19
Filing date: 2017-04-19
Publication date: 2018-11-02
Also published as: WO2018192536A1

Abstract

The present invention relates to as bruton's tyrosine kinase inhibitor heterocyclic compound and its application.Specifically, the present invention relates to compounds, pharmaceutical composition containing compound of formula I wherein shown in the Formulas I for having bruton's tyrosine kinase inhibitory activity and the compound to prepare the application in treating or preventing bruton's tyrosine kinase relevant disease or inhibiting the drug of bruton's tyrosine kinase：

Description

Pyrimido-heterocyclic compounds as Bruton's tyrosine kinase inhibitors and uses thereof

Technical Field

The present invention relates to the field of pharmaceutical chemistry; in particular, the invention relates to compounds having Bruton's tyrosine kinase inhibitory activity and uses thereof.

Background

Immune cells can be generally divided into two categories, T cells and B cells, wherein the primary role of B cells is to secrete various antibodies to help the body resist the invasion of various foreign enemies. Bruton's tyrosine kinase (Bruton's tyrosine kinase) is mainly expressed in B cells and is distributed in the lymphatic, hematopoietic and blood systems. In recent years, researches on B cells, particularly B cell non-Hodgkin lymphoma and rheumatoid arthritis show that Bruton tyrosine kinase is often abnormally expressed. Bruton's tyrosine kinase is a key kinase in the signal pathway of B cell antigen receptor (BCR), can regulate the maturation and differentiation of normal B cells, and is also closely related to various diseases of B cell lymphoid tissue disorder.

Bruton's tyrosine kinase is a member of the Tec family of non-receptor protein tyrosine kinases. The Tec family is the second 2 large family of human non-receptor kinases to the Src family, the major members of which include bruton's tyrosine kinase, bmx (etk), ITK, Tec, and txk (rlk). Bruton's tyrosine kinase was identified in 1993 as a defective protein in human X-linked agammaglobulinemia (XLA). This protein is expressed in ALL stages of B cell development (except for terminally differentiated plasma cells), bruton's tyrosine kinase is an essential gene for cell differentiation and proliferation during the transition from pre-B lymphocytes to post-B cells, and is expressed in B cell lymphomas, Acute Lymphoblastic Leukemia (ALL), and plasmacytomas. In addition, there is a small amount of expression in bone marrow cells and erythroid progenitor cells.

The Bruton's tyrosine kinase structure contains 5 major domains, namely the PH domain (Pleckstrinhomogy), the TH domain (Tec homogy), the SH3 domain (Src homogy 3), the SH2 domain (Srchology 2) and the SH1 domain (Src homogy 1), wherein the PH domain contains the transcription factor BAP-135/TFII-I and the activity down-regulator PIN1, the binding site for IBruton's tyrosine kinase, and is also responsible for mediating the action of Bruton's tyrosine kinase and the 2 nd messenger phosphatidylinositol triphosphate (PIP 3). the TH domain is adjacent to the PH domain and is composed of 80 amino acid residues, including the Bruton's tyrosine kinase motif (Zn cofactor binding site), the PKC- β binding site and the conserved domain rich proline motif the SH1 domain contains activation loop, ATP binding site, catalytic converter and fragment of the survival inhibitory fragment, including the multiple phosphorylation sites of the Bruton's tyrosine kinase (NL) in the domain, including the primary receptor phosphorylation site for activating the receptor tyrosine kinase, the initiation of the receptor cycle, the receptor cycle of the Bruton tyrosine kinase, the receptor phosphorylation process of the Bruton tyrosine kinase (NL tyrosine kinase) and the initiation of the receptor cycle.

The process of bruton's tyrosine kinase activation is complex, and an important step in this process is the migration of bruton's tyrosine kinase to the cell membrane. Some receptors on the cell membrane are activated by stimulation with the corresponding ligands, the activated receptors recruit and phosphorylate the intracellular signal transduction kinase PI3K, and the phosphorylated PI3K in turn converts PIP2 on the membrane into the 2 nd messenger PIP 3. PIP3 binds to the PH domain of Bruton's tyrosine kinase, which is then recruited to the cell membrane, Tyr-551 residues are phosphorylated by Syk and Lyn kinase, then self-phosphorylation reaction is carried out at Tyr-223 residues, so that the Bruton's tyrosine kinase with physiological activity can be combined with adaptor BLNK/SLP65 through SH2 domain, the generated compound then activates phospholipase C-gamma 2 (PLC-gamma 2), and then the cascade reaction is initiated to finally lead to the continuous calcium ion influx in the cell, and indirectly activate the downstream signal paths, such as MEK/ERK, p38MAPK and NK/SAPK paths. Bruton's tyrosine kinase gain-of-function mutations have also been identified in colorectal cancer, Acute Lymphocytic Leukemia (ALL), and Chronic Myelogenous Leukemia (CML). Thus, aberrant activation of the bruton's tyrosine kinase-dependent pathway has been shown to be closely associated with the development of a variety of tumors.

The Bruton's tyrosine kinase small-molecule inhibitor has good prospect for treating hematological malignancy and autoimmune disorder diseases. Ibrutinib (ibrutinib) is currently the most attractive target inhibitor of bruton's tyrosine kinase, has significant therapeutic effects on a variety of B cell tumors and autoimmune diseases in preclinical and clinical studies, and has been approved by the FDA in the united states for marketing for the treatment of Mantle Cell Lymphoma (MCL) and CLL. Several other compounds, such as CC-292 and ONO-4059, have also entered clinical or late clinical studies.

There is still a need in the art to develop bruton's tyrosine kinase inhibitors with high activity and specificity.

Disclosure of Invention

The invention aims to provide a Bruton's tyrosine kinase inhibitor with high activity and strong specificity and application thereof in preparing a medicament for treating Bruton's tyrosine kinase mediated diseases.

In a first aspect, the present invention provides the use of a compound of formula I, or a salt thereof, in the manufacture of a bruton's tyrosine kinase inhibitor or in the manufacture of a medicament for the treatment or prevention of a bruton's tyrosine kinase mediated disease:

in the formula,

r is hydrogen, C₁-C₃Lower alkyl, C₁-C₃Lower alkoxy, halogen (e.g., F, Cl, Br), amino, substituted amino;

x is N or CR⁵R⁶；

Y is C or O;

when the X is N, the N is N,is a double bond, and Y is C;

when X is CR⁵R⁶When the temperature of the water is higher than the set temperature,is a single bond, and Y is O; or when X is CR⁵R⁶When the temperature of the water is higher than the set temperature,is a double bond, and Y is C;

b is selected from the following group: optionally substituted (C3-C8) cycloalkyl, (C3-C8) heterocyclyl, (C6-C10) aryl, or (C5-C10) arylheterocyclyl;

R¹selected from: H. optionally substituted C₁-C₆Alkyl, NR⁷R⁸Optionally substituted C₆-C₁₀An aryl group;

R³selected from: hydrogen, optionally substituted C₁-C₁₀Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, optionally substituted C₃-C₈Cycloalkyl, optionally substituted C₁-C₁₀Alkoxy, optionally substituted aryl, optionally substituted benzyl, optionally substituted heterocyclyl, optionally substituted arylheterocyclyl, -O- (CH)_z-O-C₁-C₃An alkyl group; z is an integer from 1 to 3, preferably 1;

R⁴selected from: hydrogen, optionally substituted C₁-C₆Alkyl, nitro, amino, halogen, optionally substituted C₁-C₆Alkoxy, optionally substituted acyloxy, optionally substituted acylamino, optionally substituted acyl;

m is independently an integer from 0 to 7, preferably from 1 to 7, more preferably from 1 to 3;

R⁵and R⁶Each independently is H, or C₁-C₆Alkyl (preferably C)₁-C₃Alkyl groups);

R⁷and R⁸Each independently is H, or C₁-C₆An alkyl group.

In a preferred embodiment, B is selected from various substituted benzene rings, nitrogen-containing five-membered rings, nitrogen-containing six-membered rings or C₃-C₈A cycloalkyl group.

In a specific embodiment, B is selected from:

R⁴selected from:

in a specific embodiment, the compound is represented by formula I-1 below:

in the formula,

a is benzene ring, five-membered or six-membered heterocycle, C₃-C₈Cycloalkyl or R';

when A is R ', n is 0 and R' is selected from C₁-C₆Alkyl radical, C₁-C₆Haloalkyl or C₆-C₁₀An arylformyl group;

R²selected from: hydrogen, halogen, optionally substituted C₁-C₆Alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C₁-C₆Alkyl, CN, sulfonic acid group, aminosulfonyl, carbamoyl, carboxy, optionally substituted alkoxycarbonylOptionally substituted phenyl, optionally substituted N-alkylpiperazino, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR_aR_bOptionally substituted pyridyl; r_aAnd R_bEach independently selected from alkyl and alkenyl;

n is independently an integer of 0 to 7, preferably 1 to 7, more preferably 1 to 3;

X、Y、B、R¹、R³、R⁴and m is as defined above.

In a specific embodiment, the compound is represented by formula II-1 below:

in the formula,

B、R²、R³、R⁴m and n are as defined above;

alternatively, the compound is represented by the following formula II-2:

in the formula,

B、R²、R⁴、R⁵、R⁶m and n are as defined above;

alternatively, the compound is represented by the following formula II-3:

in the formula,

B、R¹、R²、R³、R⁴、mand n is as defined above.

In a particular embodiment of the present invention,

in the formula II-1, the compound represented by the formula,

R²selected from: hydrogen, halogen, optionally substituted C₁-C₆Alkoxy, optionally substituted pyrrolidinyl, -NR_aR_bCarbamoyl, optionally substituted amido, - (CH)₂)_o-optionally substituted N-alkylpiperazino, optionally substituted morpholinyl, optionally substituted piperidinyl, o is an integer from 0 to 2, R_aAnd R_bEach independently selected from C₁-C₃An alkyl group; wherein R is²Is not located at the 2-position of the benzene ring in which it is located;

R³selected from: hydrogen, optionally substituted C₁-C₆Alkyl, optionally substituted C₆-C₁₀Aryl, optionally substituted C₃-C₈A cycloalkyl group;

b is selected from benzene ring or five-membered ring containing nitrogen;

R⁴selected from: optionally substituted acylamino, optionally substituted acyl;

m and n are as defined above;

in the formula II-2, the compound represented by the formula,

R⁵、R⁶independently selected from H, substituted or unsubstituted C₁-C₆(preferably C)₁-C₃) An alkyl group;

b is a benzene ring;

R²selected from: optionally substituted C₁-C₆Alkyl (preferably C)₁-C₃Alkyl), optionally substituted N-alkylpiperazino, optionally substituted C₁-C₆Alkoxy (preferably C)₁-C₃Alkoxy groups);

R⁴selected from: optionally substituted amido;

m and n are as defined above.

In a second aspect, the present invention provides the use of a compound selected from the group consisting of:

in a specific embodiment, the bruton's tyrosine kinase mediated disease is cancer or and autoimmune disorders.

In specific embodiments, the cancer is selected from the group consisting of: acute Lymphocytic Leukemia (ALL), Chronic Myeloid Leukemia (CML), Mantle Cell Lymphoma (MCL), large intestine cancer; the autoimmune disorder disease includes rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis, and lupus erythematosus.

In a third aspect, the present invention provides a method of treatment or prophylaxis of bruton's tyrosine kinase mediated diseases comprising administering to a subject in need thereof a compound according to the first or second aspects of the invention or a pharmaceutical composition comprising said compound.

In a fourth aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof:

in the formula,

X、Y、B、R¹、R³、R⁴and m is as defined above;

wherein,

r is hydrogen, C₁-C₃Lower alkyl, C₁-C₃Lower alkoxy, halogen (e.g. F, Cl, Br), amino or NR^cR^dAnd R is^c、R^dIndependently selected from H, C₁-C₆Alkyl radical, C₁-C₆Haloalkyl or (C)₆-C₁₀) An arylformyl group;

and/or

R³Selected from the group consisting of: hydrogen, (C)₃-C₆) Cycloalkyl group, (C)₁-C₈) Heterocyclic group, (C)₁-C₈) Alkoxy, -O- (CH)_n-O-C₁-C₃Alkyl, benzyl, (C)₆-C₁₀) Aryl or (C)₅-C₁₀) An aromatic heterocyclic group, wherein said aryl and aromatic heterocyclic groups may be optionally substituted with one to five of the following groups: halogen, nitro, cyano, hydroxy, amino, (C)₁-C₈) Alkyl, (C)₁-C₈) Alkoxy group, (C)₃-C₆) Cycloalkyl group, (C)₆-C₁₀) Aryloxy group, (C)₅-C₁₀) Heterocyclyl, -O- (CH)_z-O-C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl oxy, C₃-C₆Heterocycloalkyloxy, amido, optionally substituted carbamoyl; z is an integer from 1 to 3, preferably 1;

and/or

R⁴Selected from the group consisting of:

in a preferred embodiment of the present invention,

r is hydrogen, C₁-C₃Lower alkyl, C₁-C₃Lower alkoxy, halogen (e.g., F, Cl, Br), amino;

or

R⁴selected from the group consisting of:

or

R³Selected from the group consisting of: hydrogen, (C)₃-C₆) Cycloalkyl group, (C)₁-C₈) Heterocyclic group, (C)₁-C₈) Alkoxy, -O- (CH)_n-O-C₁-C₃Alkyl, benzyl, (C)₆-C₁₀) Aryl or (C)₅-C₁₀) An aromatic heterocyclic group, wherein said aryl and aromatic heterocyclic groups may be optionally substituted with one to five of the following groups: halogen, nitro, cyano, hydroxy, ammoniaBase, (C)₁-C₈) Alkyl, (C)₁-C₈) Alkoxy group, (C)₃-C₆) Cycloalkyl group, (C)₆-C₁₀) Aryloxy group, (C)₅-C₁₀) Heterocyclyl, -O- (CH)_z-O-C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl oxy, C₃-C₆Heterocycloalkyloxy, amido, optionally substituted carbamoyl; z is an integer from 1 to 3, preferably 1;

R⁴selected from the group consisting of:

in a specific embodiment, the compound is represented by formula I-1 below:

in the formula,

a is R ', n is 0, and R' is selected from C₁-C₆Alkyl radical, C₁-C₆Haloalkyl or (C)₆-C₁₀) An arylformyl group;

X、Y、B、R¹、R³、R⁴and m is as defined above.

In a particular embodiment, R' is C₁-C₆Alkyl (preferably C)₁-C₃Alkyl group), C₁-C₆Haloalkyl (preferably C)₁-C₃Haloalkyl).

In a specific embodiment, R³Selected from the group consisting of:

in a specific embodiment, R³Comprises the following steps:

it is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Detailed Description

The inventors have conducted extensive and intensive studies and unexpectedly found a group of compounds having novel structures, which are capable of inhibiting Bruton's tyrosine kinase with high activity and high selectivity, and IC of Bruton's tyrosine kinase inhibitory activity of some of the compounds₅₀Values reached the nM range. The present invention has been completed based on this finding.

The present inventors have synthesized a series of candidate compounds having bruton's tyrosine kinase inhibitory activity. By carrying out structure optimization design on the obtained candidate compounds, a batch of novel pyrimidopyrimidine heterocyclic compounds with potential Bruton tyrosine kinase inhibitory activity are discovered. The activity of the obtained compounds is evaluated at the molecular level, and a plurality of compounds have Bruton tyrosine kinase inhibition activity IC₅₀Values reached the nM range.

Definition of terms

The groups to which the present invention relates have meanings conventionally understood in the art. For clarity, some groups referred to herein are defined as follows:

as used herein, "alkyl" refers to a saturated, branched or straight-chain alkyl group having a carbon chain length of 1 to 10 carbon atoms, with preferred alkyl groups including those varying in length from 2 to 8, 1 to 6, 1 to 4, 3 to 8, 1 to 3 carbon atoms. Examples of alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl and the like. The alkyl group may be substituted with 1 or more substituents, for example, with halogen or haloalkyl. For example, the alkyl group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group. The alkyl groups described herein may also be substituted with aryl groups, thereby forming, for example, benzyl groups.

Similarly, "cycloalkyl" herein refers to a substituted or unsubstituted saturated cyclic alkyl group having a carbon chain length of 3 to 10, preferably 3 to 8 carbon atoms, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.

Herein, "alkoxy" refers to an oxy group substituted with an alkyl group. Preferred alkoxy groups are alkoxy groups of 1 to 6 carbon atoms in length, more preferably 1 to 4 carbon atoms in length, and even more preferably 1 to 3 carbon atoms in length. Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, propoxy, and the like. Alkoxy groups may be substituted with 1 or more substituents, for example with halogen or haloalkyl. For example, the alkoxy group may be an alkyl group substituted with 1 to 4 fluorine atoms, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.

As used herein, "alkenyl" generally refers to a monovalent hydrocarbon group having at least one double bond, generally containing 2 to 8 carbon atoms, preferably 2 to 6 carbon atoms, and may be straight or branched. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, hexenyl, and the like.

As used herein, "alkynyl" generally refers to a monovalent hydrocarbon group having at least one triple bond, generally containing 2 to 8 carbon atoms, preferably 2 to 6 carbon atoms, more generally 2 to 4 carbon atoms, and may be straight or branched. Examples of alkenyl groups include ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, hexynyl, and the like.

Herein, "halogen" means fluorine, chlorine, bromine or iodine.

As used herein, "aryl" means a monocyclic, bicyclic or tricyclic aromatic group having 6 to 14 carbon atoms, and includes phenyl, naphthyl, phenanthryl, anthryl, indenyl, fluorenyl, tetralinyl, indanyl and the like. Aryl groups may be optionally substituted with 1-5 (e.g., 1, 2,3, 4, or 5) substituents selected from: halogen, C1-4 aldehyde, C1-6 alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen-substituted alkyl (e.g., trifluoromethyl), halogen-substituted alkoxy (e.g., trifluoromethoxy), carboxy, C1-4 alkoxy, ethoxyformyl, N (CH3) and C1-4 acyl, and the like, heterocyclic group, heteroaryl group, and the like.

As used herein, "heterocyclyl" includes, but is not limited to, 5-or 6-membered heterocyclic groups containing 1-3 heteroatoms selected from O, S or N, including, but not limited to, furyl, thienyl, pyrrolyl, pyrrolidinyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, morpholinyl, and the like.

As used herein, "arylheterocyclyl" means a ring system containing 5 to 14 ring atoms and having 6, 10, or 14 electrons in common in the ring system. And the ring atoms contained are carbon atoms and 1 to 3 heteroatoms selected from oxygen, nitrogen and sulfur. Useful aryl heterocyclic groups include piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, including but not limited to 2-pyridyl, 3-pyridyl and 4-pyridyl, pyrazinyl, pyrimidinyl and the like.

The aryl heterocyclic group may be optionally substituted with 1 to 5 (e.g., 1, 2,3, 4, or 5) substituents selected from the group consisting of: halogen, C1-4 aldehyde, C1-6 straight or branched chain alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (e.g., trifluoromethyl), halogen substituted alkoxy (e.g., trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxyformyl, N (CH3), and C1-4 acyl.

As used herein, "acyloxy" refers to a group of the formula "-O-C (O) -R", wherein R may be selected from alkyl, alkenyl (e.g., C1-6 or C1-3 alkenyl), and alkynyl. The R may be optionally substituted.

As used herein, "amido" refers to a group of the formula "-R '-NH-C (O) -R", wherein R' may be selected from hydrogen or alkyl, and R may be selected from alkyl, alkenyl (e.g., C1-6 or C1-3 alkenyl), alkynyl, or NR_cR_dSubstituted alkyl, by NR_cR_dSubstituted alkenyl and NR_cR_dSubstituted alkynyl, alkyl substituted by halogen, alkenyl substituted by cyano, wherein R_cAnd R_dCan be selected from alkyl and alkenyl.

As used herein, acyl refers to the radical of an organic or inorganic oxyacid having one or more hydroxyl groups removed and is represented by the general formula R-C (O) -wherein R may be selected from alkyl, alkenyl (e.g., C1-6 or C1-3 alkenyl), alkynyl, or NR_cR_dSubstituted alkyl, by NR_cR_dSubstituted alkenyl and NR_cR_dSubstituted alkynyl, alkyl substituted by halogen, alkenyl substituted by cyano, wherein R_cAnd R_dCan be selected from alkyl and alkenyl.

As used herein, "arylformyl" refers to a group formed by an aryl group, e.g., (C6-C10) aryl, attached to the main structure of the compound through a formyl group.

Herein, "optionally substituted" means that the substituent group it modifies may be optionally substituted with 1 to 5 (e.g., 1, 2,3, 4, or 5) substituents selected from: halogen, C1-4 aldehyde, C1-6 straight or branched chain alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, halogen substituted alkyl (e.g., trifluoromethyl), halogen substituted alkoxy (e.g., trifluoromethoxy), carboxyl, C1-4 alkoxy, ethoxyformyl, N (CH3), and C1-4 acyl.

Compounds of the invention and uses thereof

For the purpose of the present invention, the present invention provides the use of a compound represented by the following formula I or a salt thereof for the preparation of a bruton's tyrosine kinase inhibitor or for the preparation of a medicament for the treatment or prevention of a bruton's tyrosine kinase mediated disease:

in the formula,

x is N or CR⁵R⁶；

Y is C or O;

when the X is N, the N is N,is a double bond, and Y is C;

b is selected from the following group: optionally substituted (C3-C8) cycloalkyl, (C3-C8) heterocyclyl, (C6-C10) aryl, or (C5-C10) arylheterocyclyl; r¹Selected from: H. optionally substituted C₁-C₆Alkyl, NR⁷R⁸Optionally substituted C₆-C₁₀An aryl group; r³Selected from: hydrogen, optionally substituted C₁-C₁₀Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, optionally substituted C₃-C₈Cycloalkyl, optionally substituted C₁-C₁₀Alkoxy, optionally substituted aryl, optionally substituted benzyl, optionally substituted heterocyclyl, optionally substituted arylheterocyclyl, -O- (CH)_z-O-C₁-C₃An alkyl group; z is an integer from 1 to 3, preferably 1; r⁴Selected from: hydrogen, optionally substituted C₁-C₆Alkyl, nitro, amino, halogen, optionally substituted C₁-C₆Alkoxy, optionally substituted acyloxy, optionally substituted acylamino, optionally substituted acyl; m is independently an integer from 0 to 7, preferably from 1 to 7, more preferably from 1 to 3; r⁵And R⁶Each independently is H, or C₁-C₆Alkyl (preferably C)₁-C₃Alkyl groups); r⁷And R⁸Each independently is H, or C₁-C₆An alkyl group.

Further, B may be selected from:

R⁴selected from:

further, the compound of the present invention may be represented by the following formula I-1:

in the formula,

R²selected from: hydrogen, halogen, optionally substituted C₁-C₆Alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C₁-C₆Alkyl, CN, sulfonic acid, aminosulfonyl, carbamoyl, carboxy, optionally substituted alkoxycarbonyl, optionally substituted phenyl, optionally substituted N-alkylpiperazino, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR_aR_bOptionally substituted pyridyl; r_aAnd R_bEach independently selected from alkyl and alkenyl;

X、Y、B、R¹、R³、R⁴and m is as defined above.

Still further, the compounds of the present invention are represented by the following formula II-1:

in the formula,

B、R²、R³、R⁴m and n are as defined above;

alternatively, the compound is represented by the following formula II-2:

in the formula,

B、R²、R⁴、R⁵、R⁶m and n are as defined above;

alternatively, the compound is represented by the following formula II-3:

in the formula,

B、R¹、R²、R³、R⁴m and n are as defined above.

In a still further aspect of the present invention,

in the formula II-1, the compound represented by the formula,

b is selected from benzene ring or five-membered ring containing nitrogen;

m and n are as defined above;

in the formula II-2, the compound represented by the formula,

b is a benzene ring;

R⁴selected from: optionally substituted amido;

m and n are as defined above.

In a particular embodiment, the present invention provides the use of a compound selected from the group consisting of:

the above table shows the structure of the compounds of the invention and their bruton's tyrosine kinase inhibitory activity, wherein:

IC of Compounds whose Activity is designated "A₅₀≤10nM；

IC of Compounds Activity designated "B₅₀Is 10<IC₅₀≤100nM；

IC of Compounds whose Activity is designated "C₅₀Is 100<IC₅₀≤1000nM；

IC of Compounds whose Activity is designated "D₅₀Is 1000nM<IC₅₀。

On the basis that the compound disclosed by the invention has the Bruton tyrosine kinase inhibition activity, the invention provides a pharmaceutical composition for inhibiting Bruton tyrosine kinase, which comprises a therapeutically effective amount of the compound disclosed by the invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

Examples of pharmaceutically acceptable salts of the compounds of the present invention include, but are not limited to, inorganic and organic acid salts, such as hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate salts; and inorganic and organic base salts formed with bases such as sodium hydroxy, TRIS (hydroxymethyl) aminomethane (TRIS, tromethamine) and N-methylglucamine.

Although the requirements vary from person to person, the skilled person can determine the optimal dosage of each active ingredient in the pharmaceutical composition of the invention. Typically, the compounds of the present invention, or pharmaceutically acceptable salts thereof, are administered orally to a mammal daily in an amount of from about 0.0025 to 50 mg/kg body weight. But preferably about 0.01 to 10 mg per kg is administered orally. For example, a unit oral dosage may include from about 0.01 to 50mg, preferably from about 0.1 to 10 mg, of a compound of the present invention. A unit dose may be administered one or more times daily in one or more tablets, each tablet containing from about 0.1 to 50mg, conveniently from about 0.25 to 10 mg, of a compound of the invention or a solvate thereof.

The pharmaceutical compositions of the present invention may be formulated in a form suitable for various routes of administration, including but not limited to, administration by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal or topical routes for the treatment of tumors and other diseases. The amount administered is an amount effective to ameliorate or eliminate one or more symptoms. For the treatment of a particular disease, an effective amount is an amount sufficient to ameliorate or in some way reduce the symptoms associated with the disease. Such amounts may be administered as a single dose or may be administered according to an effective treatment regimen. The amount administered may be sufficient to cure the disease, but is generally administered to ameliorate the symptoms of the disease. Repeated administration is generally required to achieve the desired improvement in symptoms. The dosage of the drug will depend on the age, health and weight of the patient, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.

The pharmaceutical preparation of the present invention can be administered to any mammals as long as they can obtain the therapeutic effects of the compound of the present invention. Of these mammals, the most important is human.

The compounds of the present invention or pharmaceutical compositions thereof are useful in the treatment of various diseases mediated by bruton's tyrosine kinase. Herein, the bruton's tyrosine kinase mediated disease is cancer or an autoimmune disease; wherein the cancer comprises a hematologic malignancy or a solid tumor, such as: acute Lymphocytic Leukemia (ALL), Chronic Myeloid Leukemia (CML), Mantle Cell Lymphoma (MCL), large intestine cancer; the autoimmune disease includes rheumatoid arthritis, resistance to organ transplant rejection, resistance to psoriasis or lupus erythematosus.

The pharmaceutical preparations of the present invention can be manufactured in a known manner. For example, by conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. In the manufacture of oral formulations, solid excipients and active compounds may be combined, optionally grinding the mixture. If desired or necessary after addition of suitable amounts of auxiliaries, the granulate mixture is processed to give tablets or dragee cores.

Suitable adjuvants are, in particular, fillers, for example sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations or calcium phosphates, such as tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch pastes, including corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone. If desired, disintegrating agents such as the starches mentioned above, as well as carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added. Adjuvants are, in particular, flow regulators and lubricants, for example silica, talc, stearates, such as calcium magnesium stearate, stearic acid or polyethylene glycol. If desired, a suitable coating resistant to gastric juices can be provided to the tablet core. For this purpose, concentrated saccharide solutions can be used. This solution may contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. For the preparation of coatings resistant to gastric juices, suitable cellulose solutions can be used, for example cellulose acetate phthalate or hydroxypropylmethyl cellulose phthalate. Dyes or pigments may be added to the coating of the tablet or lozenge core. For example, for identifying or for characterizing combinations of active ingredient doses.

Based on the above compounds and pharmaceutical compositions, the present invention further provides a method for treating or preventing bruton's tyrosine kinase mediated diseases, which comprises administering to a subject in need thereof a compound or pharmaceutical composition of the present invention. The method of administration includes, but is not limited to, various methods of administration known in the art, and may be determined based on the actual condition of the patient. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal, or topical routes of administration.

In a particular embodiment, the present invention provides a compound of formula I or a salt thereof, having a novel structure:

in the formula,

X、Y、B、R¹、R³、R⁴and m is as defined above;

wherein,

and/or

R³Selected from the group consisting of: hydrogen, (C)₃-C₆) Cycloalkyl group, (C)₁-C₈) Heterocyclic group, (C)₁-C₈) Alkoxy, -O- (CH)_n-O-C₁-C₃Alkyl, benzyl, (C)₆-C₁₀) Aryl or (C)₅-C₁₀) Aromatic heterocyclic group, wherein said aromatic group and aromatic heterocyclic groupOptionally substituted with one to five of the following groups: halogen, nitro, cyano, hydroxy, amino, (C)₁-C₈) Alkyl, (C)₁-C₈) Alkoxy group, (C)₃-C₆) Cycloalkyl group, (C)₆-C₁₀) Aryloxy group, (C)₅-C₁₀) Heterocyclyl, -O- (CH)_z-O-C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl oxy, C₃-C₆Heterocycloalkyloxy, amido, optionally substituted carbamoyl; z is an integer from 1 to 3, preferably 1;

and/or

R⁴Selected from the group consisting of:

in a preferred embodiment, R, R as described above³And R⁴Can be combined at will. For example, the following combinations may be possible:

or

R⁴selected from the group consisting of:

or

R⁴selected from the group consisting of:

in a specific embodiment, the structurally novel compounds of the present invention are represented by the following formula I-1:

in the formula,

X、Y、B、R¹、R³、R⁴and m is as defined above.

Further, R' may be C₁-C₆Alkyl (preferably C)₁-C₃Alkyl group), C₁-C₆Haloalkyl (preferably C)₁-C₃Haloalkyl).

In a preferred embodiment, R³May be selected from the group consisting of:

more preferably

In specific embodiments, the substituents in the general formulae of the present invention are each the corresponding group in any of the specific compounds disclosed herein.

The invention has the advantages that:

1. the compound has excellent inhibitory activity on Bruton's tyrosine kinase;

2. the compound of the invention has high selectivity to Bruton tyrosine kinase; and

3. the compound lays a foundation for developing a medicine capable of inhibiting Bruton's tyrosine kinase with high activity and high selectivity, has great industrialization and commercialization prospects and market values, and has remarkable economic benefit.

The technical solution of the present invention will be further described with reference to the following specific embodiments, but the following embodiments are not intended to limit the present invention, and all of the various application methods adopted according to the principles and technical means of the present invention belong to the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.

Materials and methods

1. Bruton's tyrosine kinase inhibition activity detection method

1.1 expression of Bruton's tyrosine kinase recombinant proteins

1) Construction of PfastBac 1-Bruton tyrosine kinase vector

Carrying out PCR amplification on a Bruton tyrosine kinase target fragment (M1-S695), carrying out double enzyme digestion on a PCR product and a vector PfastBac1 by using BamHI and XhoI, connecting enzyme digestion products, transforming the enzyme digestion products into DH5 α competent cells, selecting a single clone, and finally obtaining a recombinant plasmid pFastBac 1-Bruton tyrosine kinase with a correct sequence through sequencing verification.

2) Acquisition of baculovirus

Transposing the constructed plasmid to DH10Bac competent cells for blue-white screening, selecting a monoclonal with successful transposition, extracting bacmid after shaking bacteria, and identifying the bacmid through bacteria liquid PCR. Sf9 cells were transfected with bacmid that identified the correct plasmid, resulting in P1, P2 and higher titers of P3 virus strains, respectively.

3) Bruton's tyrosine kinase protein expression and identification

Sf9 cells were cultured to logarithmic phase (about 2X 10)⁶One cell/mL), the P3 strain having a high titer was added to a culture medium containing Sf9 cells grown in log phase, cultured at 27 ℃ for 3 days, centrifuged at 500 × g for 5min, the supernatant was discarded, harvested, and stored at-80 ℃. Protein expression was then detected by immunoblotting (Western Blot).

1.2 purification of Bruton's tyrosine kinase recombinant proteins

The pellet of cells expressed by the P3 strain was collected by centrifugation at 1790rpm at room temperature. The lysate used for lysing the cells was 250mM NaCl, 0.25% NP-40, and 50mM CHES (pH 9.0). The cells were disrupted by a high-pressure cell disrupter, and then centrifuged at 12000rpm at 4 ℃ for 45min to collect the supernatant. And (3) adding the supernatant into a Ni-NTA chromatographic column, eluting the target protein by using an imidazole concentration gradient method, and collecting the eluted protein solution. And concentrating the eluent containing the target protein, and changing the eluent to the lowest imidazole concentration. The solution was dialyzed against TEV enzyme at 4 ℃ for 16h, and the solution was re-run through a Ni-NTA column, and the His-Tag-free flow-through was collected in a buffer of 200mM NaCl,20mM CHES (pH 9.0), 1mM TCEP. Finally, the purified protein was separated by HiTrap Superdex75 molecular sieve using 100mM NaCl,10mM Tris-HCl pH 8.5, 1mM CEPT.

1.3 molecular level screening of Bruton's tyrosine kinase inhibitors

The molecular level screening experiment of the Bruton tyrosine kinase inhibitor selects Z-Assay Kit (PV 3190). The experimental method comprises: diluting a compound to be detected in a concentration gradient manner, adding 2.5 mu L of Test Compounds into a 384-hole plate, adding 5 mu L of Bruton's tyrosine Kinase Kinase/Peptide Substrate Mixture and 2.5 mu L of ATP Solution into each group of three parallel controls, oscillating for 30s, uniformly mixing, and incubating for 1h at room temperature; adding 5 mu of LDevelment Solution, oscillating for 30s, mixing uniformly, and incubating for 1h at room temperature; then 5 mul of Stop Reagent is added, the mixture is oscillated for 30s and mixed evenly, a fluorescence signal is detected by using an enzyme-labeling instrument, the excitation wavelength is 400nm, and the emission wavelength is 445nm and 520nm respectively. The inhibition of the compounds at 7 concentration gradients was determined and the IC of each compound was calculated by Origin 8.0 fitting of the curve₅₀The value is obtained.

The synthesis of the 7(8H) -pteridinone compounds of the invention is shown below:

reagents and conditions: (a) amine, DIPEA,1, 4-dioxane, r.t.; (b) ArNH2, DIPEA,1, 4-dioxane, r.t.; (c) Pd/C, H2, EtOH; (d) r2 cooet, HOAc, EtOH, reflux.

In the above preparation process, R¹、R²、R³、R⁴、R⁵Reference is made to the corresponding group definitions above. The compounds of the present invention can be prepared by those skilled in the art according to the actual preparation needs, using various starting compounds conventionally obtained in the art as starting materials.

Example 1

The specific synthesis method of the steps a-d is as follows:

synthesis of tert-butyl (4- (2-chloro-5-nitropyrimidine-4-amino) phenyl) carbamate

Weighing 2, 4-dichloro-5-nitropyrimidine (190mg, 0.98mmol), placing the weighed mixture in a 25mL round-bottom flask, adding 6mL of 1, 4-dioxane, stirring at room temperature, dissolving (4-aminophenyl) carbamic acid tert-butyl ester (200mg, 0.96mmol) and N, N-diisopropylethylamine (137mg, 1.06mmol) in 4mL of 1, 4-dioxane, dropwise adding the mixture into the reaction solution, continuing to stir at room temperature for 1 hour after the dropwise adding is completed, and tracking by TLC until the raw materials are completely converted. The solvent was removed by rotary evaporation and the crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate 10:1, v/v) to give tert-butyl (4- (2-chloro-5-nitropyrimidin-4-amino) phenyl) carbamate as an orange solid 301mg, 82% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.38(s,1H),9.47(s,1H),9.12(s,1H),7.49(d,J＝8.4Hz,2H),7.39(d,J＝8.4Hz,2H),1.49(s,9H)。

Synthesis of tert-butyl (4- (2- (4-methoxyphenylamino) -5-nitropyrimidin-4-amino) phenyl) carbamate

Tert-butyl (4- (2-chloro-5-nitropyrimidine-4-amino) phenyl) carbamate (50mg, 0.14mmol), p-anisidine (17mg, 0.14mmol), N-diisopropylethylamine (18mg, 0.18mmol) were weighed into a 10mL round-bottomed flask, 5mL of 1, 4-dioxane was added, stirring was carried out at room temperature for 4 hours, and TLC was followed until the starting material was completely converted. The solvent was removed by rotary evaporation and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate 4:1, v/v) to give (4- (2- (4-methoxyphenylamino) -5-nitropyrimidine-4-amino) phenyl) carbamic acid tert-butyl ester as a yellow solid 51mg in 82% yield.

¹H NMR(400MHz,DMSO-d6):δ10.30(s,1H),10.26(s,1H),9.45(s,1H),9.04(s,1H),7.49(d,J＝8.8Hz,2H),7.45(d,J＝8.8Hz,2H),7.40(d,J＝8.6Hz,2H),6.75(d,J＝8.6Hz,2H),3.73(s,3H),1.50(s,9H)。

Synthesis of tert-butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidin-4-amino) phenyl) carbamate

Tert-butyl (4- (2- (4-methoxyphenylamino) -5-nitropyrimidin-4-amino) phenyl) carbamate (45mg, 0.10mmol) was weighed into a 50mL round-bottomed flask, 20mL of ethanol and 5mg of palladium on carbon (10% Pd) were added, and hydrogen was introduced and the mixture was stirred at room temperature overnight. After the reaction was completed, suction filtration was performed, the filtrate was spin-dried, and the crude product was purified by silica gel column chromatography (dichloromethane/methanol ═ 5:1, v/v) to give tert-butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidin-4-amino) phenyl) carbamate as a pale pink solid (30mg, yield 83%).

¹H NMR(400MHz,DMSO-d6):δ9.23(s,1H),8.42(s,1H),8.10(s,1H),7.62(d,J＝9.2Hz,2H),7.56(s,1H),7.53(d,J＝9.2Hz,2H),7.40(d,J＝8.8Hz,2H),6.77(d,J＝8.8Hz,2H),3.70(s,3H),1.48(s,9H)。

Synthesis of tert-butyl (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) carbamate

Tert-butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidin-4-amino) phenyl) carbamate (30mg,0.07mmol) was weighed into a 10mL round-bottomed flask, and 0.29mL glacial acetic acid, 5mL absolute ethanol, and then ethyl glyoxylate (50% in toluene) (16mg, 0.08mmol) were added, and the mixture was heated to reflux and stirred overnight. After the reaction is finished, a solid is separated out, is filtered, and a filter cake is washed by ethanol, ammonia water and deionized water and is dried. 18mg of (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) carbamic acid tert-butyl ester was obtained as a yellow solid in a yield of 76%.

¹H NMR(400MHz,DMSO-d₆):δ10.08(s,1H),9.64(s,1H),8.84(s,1H),8.03(s,1H),7.65(d,J＝8.4Hz,2H),7.30-7.28(m,4H),6.61(br,2H),3.67(s,3H),1.52(s,9H)。

Synthesis of 8- (4-aminophenyl) -2- (4-methoxyphenyl) -7(8H) -pteridinone (SEQ ID NO: 1)

Tert-butyl (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) carbamate (18mg,0.04mmol) was weighed into a 5mL round bottom flask, 2mL dichloromethane was added, stirring was performed at 0 ℃ and 0.5mL trifluoroacetic acid was added. Stirring was then continued at 0 ℃ for 1 hour and at room temperature for 1 hour. After the reaction was completed, saturated sodium bicarbonate solution was added to neutralize the solution until it was basic, dichloromethane was used for extraction (3 × 50mL), the organic phase was washed with deionized water, saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was spin dried. 14mg of 8- (4-aminophenyl) -2- (4-methoxyphenyl) -7(8H) -pteridinone are obtained as a yellow solid in 99% yield. mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.04(br,1H),8.81(s,1H),8.00(s,1H),7.40(d,J＝7.6Hz,2H),6.98(d,J＝8.4Hz,2H),6.73(d,J＝8.4Hz,2H),6.67(br,2H),5.44(s,2H),3.70(s,3H)。¹³C NMR(100MHz,DMSO-d₆):δ159.19,158.53,157.17,154.95,151.76,149.66,146.68,133.17,129.22,122.66,121.04,120.70,114.37,113.87,55.55.HPLC purity:97.6％,Retention time＝12.59min.HRMS(ESI):exact mass calcd for C₁₉H₁₇N₆O₂[M+H]⁺,361.1413,found 361.1414。

The following compounds were synthesized according to the procedure described above:

8- (3-aminophenyl) -2- (4-methoxyphenyl) -7(8H) -pteridinone (SEQ ID NO: 2)

Yellow solid, yield 86%, mp 270.5-270.9 ℃.

¹H NMR(400MHz,DMSO-d6):δ10.06(br,1H),8.83(s,1H),8.01(s,1H),7.41(d,J＝8.0Hz,2H),7.22(t,J＝8.0Hz,1H),6.75(d,J＝7.6Hz,1H),6.67(br,2H),6.53(s,1H),6.48(d,J＝7.6Hz,1H),5.35(s,2H),3.69(s,3H).HPLC purity:94.4％,Retention time＝12.90min.HRMS(ESI):exact mass calcd for C19H17N6O2[M+H]+,361.1413,found361.1413.

2- (3-Aminophenyl) -8- (4-methoxyphenyl) -7(8H) -pteridinone (SEQ ID NO: 3)

Yellow solid, yield 85%, mp 244.5-245.4 ℃.

¹H NMR(400MHz,DMSO-d6):δ9.91(s,1H),8.85(s,1H),8.04(s,1H),7.35(d,J＝8.8Hz,2H),7.16(d,J＝8.8Hz,2H),6.68-6.65(m,3H),6.16(d,J＝7.2Hz,1H),4.63(s,2H),3.86(s,3H).HPLC purity:98.9％,Retention time＝12.57min.HRMS(ESI):exact masscalcd for C19H17N6O2[M+H]+,361.1413,found 361.1411.

2- (4-Aminophenyl) -8- (4-methoxyphenyl) -7(8H) -pteridinone (SEQ ID NO: 4)

Yellow solid, 88% yield, mp 281.5-282.3 ℃.

¹H NMR(400MHz,DMSO-d6):δ9.87(s,1H),8.77(s,1H),7.97(s,1H),7.32(d,J＝8.8Hz,2H),7.13(d,J＝8.8Hz,2H),7.08(br,2H),6.24(br,2H),4.84(s,2H),3.88(s,3H).HPLC purity:95.2％,Retention time＝11.91min.HRMS(ESI):exact mass calcd forC19H17N6O2[M+H]+,361.1413,found 361.1417.

N- (3- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 6)

Yellow solid, 74% yield, mp 260.9-261.5 ℃.

¹H NMR(400MHz,DMSO-d6):δ10.42(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.84(d,J＝8.0Hz,1H),7.78(s,1H),7.56(t,J＝8.0Hz,1H),7.31(br,2H),7.13(d,J＝8.0Hz,1H),6.58(br,2H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.6Hz,1H),5.77(dd,J＝10.4,1.6Hz,1H),3.65(s,3H).HPLC purity:97.0％,Retention time＝13.11min.HRMS(ESI):exact mass calcd for C22H19N6O3[M+H]+,415.1519,found415.1516.

N- (4- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) propanamide (SEQ ID NO. 7)

Yellow solid, yield 78%, mp 258.5-259.1 ℃.

¹H NMR(400MHz,DMSO-d6):δ10.15(s,1H),10.08(br,1H),8.85(s,1H),8.04(s,1H),7.80(d,J＝8.4Hz,2H),7.35-7.33(m,4H),6.61(br,2H),3.67(s,3H),2.41(q,J＝7.6Hz,2H),1.14(t,J＝7.6Hz,3H).HPLC purity:99.3％,Retention time＝13.05min.HRMS(ESI):exact mass calcd for C22H21N6O3[M+H]+,417.1675,found417.1674.

N- (3- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) propanamide (SEQ ID NO. 8)

Yellow solid, yield 80%, mp 298.9-299.4 ℃.

1H NMR(400MHz,DMSO-d6):δ10.13(s,1H),10.09(s,1H),8.85(s,1H),8.04(s,1H),7.74(d,J＝8.0Hz,1H),7.71(s,1H),7.53(t,J＝8.0Hz,1H),7.31(br,2H),7.07(d,J＝8.0Hz,1H),6.59(br,2H),3.67(s,3H),2.33(q,J＝7.6Hz,2H),1.07(t,J＝7.6Hz,3H).HPLCpurity:98.1％,Retention time＝13.13min.HRMS(ESI):exact mass calcd forC22H21N6O3[M+H]+,417.1675,found 417.1678.

N- (3- ((8- (4-methoxyphenyl) -7-oxo-8 (7H) pteridin-2-yl) phenyl) acrylamide (SEQ ID NO: 9)

Yellow solid, 73% yield, mp 243.3-244.0 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.17(s,1H),10.01(s,1H),8.89(s,1H),8.07(s,1H),7.63(br,1H),7.33(d,J＝8.8Hz,2H),7.27(d,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),7.11(d,J＝8.8Hz,2H),6.89(br,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.74(dd,J＝10.2,1.8Hz,1H),3.85(s,3H).HPLC purity:99.0％,Retention time＝12.91min.HRMS(ESI):exact mass calcd for C₂₂H₁₉N₆O₃[M+H]⁺,415.1519,found415.1519.

N- (4- ((8- (4-methoxyphenyl) -7-oxo-8 (7H) pteridin-2-yl) phenyl) acrylamide (SEQ ID NO: 10)

Yellow solid, 77% yield, mp 275.3-276.4 ℃.

1H NMR(400MHz,DMSO-d6):δ10.19(br,1H),10.03(s,1H),8.87(s,1H),8.05(s,1H),7.36-7.34(m,6H),7.16(d,J＝8.4Hz,2H),6.41(dd,J＝17.0,10.2Hz,1H),6.23(dd,J＝17.0,1.6Hz,1H),5.72(dd,J＝10.2,1.6Hz,1H),3.92(s,1H).HPLC purity:96.9％,Retention time＝12.75min.HRMS(ESI):exact mass calcd for C22H19N6O3[M+H]+,415.1519,found 415.1524。

Phenyl 4- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) acrylate (SEQ ID NO: 11)

Yellow solid, yield 69%, mp 257.2-258.0 ℃.

¹H NMR(400MHz,DMSO-d6):δ10.15(s,1H),8.87(s,1H),8.06(s,1H),7.51(d,J＝8.8Hz,2H),7.45(d,J＝8.8Hz,2H),7.31(br,2H),6.69(br,2H),6.60(dd,J＝17.2,1.6Hz,1H),6.51(dd,J＝17.2,9.9Hz,1H),6.22(dd,J＝9.9,1.6Hz,1H),3.67(s,3H).HPLCpurity:97.8％,Retention time＝15.64min.HRMS(ESI):exact mass calcd forC22H18N5O4[M+H]+,416.1359,found 416.1359.

(E) -4- (dimethylamino) -N- (4- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) -2-butenamide (SEQ ID NO. 12)

Yellow solid, yield 55%, mp 273.5-274.3 ℃.

1H NMR(400MHz,DMSO-d6):δ10.45(s,1H),10.10(br,1H),8.85(s,1H),8.05(s,1H),7.87(d,J＝8.8Hz,2H),7.37(d,J＝8.8Hz,2H),7.30(br,2H),6.82(td,J＝15.4,6.0Hz,1H),6.60(br,2H),6.40(d,J＝15.4Hz,1H),3.63(s,3H),3.27(d,J＝5.2Hz,2H),2.33(s,6H).HPLC purity:97.7％,Retention time＝10.47min.HRMS(ESI):exact masscalcd for C25H26N7O3[M+H]+,472.2097,found 472.2095.

(E) -4- (dimethylamino) -N- (3- (2- ((4-methoxyphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) -2-butenamide (SEQ ID NO. 13)

Yellow solid, yield 47%, mp 185.1-185.9 ℃.

1H NMR(400MHz,DMSO-d6):δ10.33(s,1H),10.08(br,1H),8.86(s,1H),8.05(s,1H),7.83(d,J＝8.0Hz,1H),7.78(s,1H),7.55(t,J＝8.0Hz,1H),7.32(br,2H),7.11(d,J＝8.0Hz,1H),6.74(td,J＝15.2,5.6Hz,1H),6.59(br,2H)6.30(d,J＝15.2Hz,1H),3.66(s,3H),3.06(d,J＝5.6Hz,2H),2.17(s,6H).HPLC purity:98.3％,Retention time＝10.74min.HRMS(ESI):exact mass calcd for C25H24N7O3[M+H]+,472.2097,found472.2094.

(E) -4- (dimethylamino) -N- (4- (7-oxo-2- (phenylamino) -8(7H) pteridinyl) phenyl) -2-butenamide (SEQ ID NO. 14)

Yellow solid, yield 53%, mp 225.3-226.1 ℃.

1H NMR(400MHz,DMSO-d6):δ10.37(s,1H),10.19(br,1H),8.90(s,1H),8.08(s,1H),7.87(d,J＝8.4Hz,2H),7.42(d,J＝7.6Hz,2H),7.38(d,J＝8.4Hz,2H),7.03(br,1H),6.88(t,J＝7.6Hz,1H),6.82(td,J＝15.4,5.6Hz,1H),6.37(d,J＝15.4Hz,1H),3.14(d,J＝5.6Hz,2H),2.24(s,6H).HPLC purity:98.2％,Retention time＝10.55min.HRMS(ESI):exact mass calcd for C24H24N7O2[M+H]+,442.1991,found 442.1989.

(E) -4- (dimethylamino) -N- (3- (7-oxo-2- (phenylamino) -8(7H) pteridinyl) phenyl) -2-butenamide (SEQ ID NO. 15)

Yellow solid, yield 46%, mp 183.8-184.3 ℃.

1H NMR(400MHz,DMSO-d6):δ10.32(s,1H),10.17(s,1H),8.90(s,1H),8.08(s,1H),7.81-7.79(m,2H),7.55(t,J＝8.0Hz,1H),7.41(d,J＝7.2Hz,2H),7.12(d,J＝8.0Hz,1H),7.01(br,2H),6.87(t,J＝7.2Hz,1H),6.73(td,J＝15.2,5.6Hz,1H),6.28(d,J＝15.2Hz,1H),3.05(d,J＝5.6Hz,2H),2.16(s,6H).HPLC purity:98.2％,Retention time＝10.85min.HRMS(ESI):exact mass calcd for C24H24N7O2[M+H]+,442.1991,found442.1996.

N- (4- (7-oxo-2-phenylamino) -8(7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 16)

Yellow solid, yield 79%, mp >300 ℃.

1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.19(br,1H),8.90(s,1H),8.09(s,1H),7.88(d,J＝8.4Hz,2H),7.41-7.38(m,4H),7.03(br,2H),6.88(t,J＝7.2Hz,1H),6.53(dd,J＝16.8,10.4Hz,1H),6.35(dd,J＝16.8,1.6Hz,1H),5.84(dd,J＝10.4,1.6Hz,1H).HPLC purity:96.1％,Retention time＝12.68min.HRMS(ESI):exact mass calcd forC21H17N6O2[M+H]+,385.1413,found 385.1405.

N- (3- (7-oxo-2-phenylamino) -8(7H) pteridinyl) phenyl) acrylamide (No. 17)

Yellow solid, 74% yield, mp 270.1-270.9 ℃.

1H NMR(400MHz,DMSO-d6):δ10.42(s,1H),10.19(s,1H),8.91(s,1H),8.09(s,1H),7.84-7.81(m,2H),7.57(t,J＝8.0Hz,1H),7.41(br,2H),7.15(d,J＝7.6Hz,1H),7.02(br,2H),6.87(t,J＝7.6Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.6Hz,1H),5.77(dd,J＝10.4,1.6Hz,1H).HPLC purity:96.8％,Retention time＝13.27min.HRMS(ESI):exact mass calcd for C21H17N6O2[M+H]+,385.1413,found385.1413.

N- (4- (2- ((4-chlorophenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 18)

Yellow solid, yield 81%, mp >300 ℃.

¹H NMR(400MHz,DMSO-d6):δ10.46(s,1H),10.34(s,1H),8.92(s,1H),8.11(s,1H),7.88(d,J＝8.8Hz,2H),7.41-7.36(m,4H),7.06(br,2H),6.53(dd,J＝16.8,10.4Hz,1H),6.36(dd,J＝16.8,1.6Hz,1H),5.84(dd,J＝10.4,1.6Hz,1H).HPLC purity:94.3％,Retention time＝14.43min.HRMS(ESI):exact mass calcd for C21H16N6O2Cl[M+H]+,419.1023,found 419.1031.

N- (3- (2- ((4-chlorophenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 19)

Yellow solid, yield 78%, mp 259.1-259.8 ℃.

1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.34(br,1H),8.93(s,1H),8.11(s,1H),7.84(s,1H),7.81(d,J＝8.4Hz,1H),7.59(t,J＝8.0Hz,1H),7.43(d,J＝7.2Hz,2H),7.15(d,J＝7.6Hz,1H),6.46(dd,J＝16.8,10.4Hz,1H),6.26(dd,J＝16.8,1.8Hz,1H),5.77(dd,J＝10.12,1.8Hz,1H).HPLC purity:97.4％,Retention time＝13.83min.HRMS(ESI):exact mass calcd for C21H16N6O2Cl[M+H]+,419.1023,found 419.1027.

N- (4- (2- ((4-Morpholphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 20)

Red solid, yield 63%, mp >300 ℃.

1H NMR(400MHz,DMSO-d6):δ10.44(s,1H),10.00(s,1H),8.82(s,1H),8.02(s,1H),7.88(d,J＝8.0Hz,1H),7.36(d,J＝8.4Hz,1H),7.22(br,2H),6.59(br,2H),6.52(dd,J＝17.2,10.2Hz,1H),6.33(d,J＝17.2Hz,1H),5.85(d,J＝10.2Hz,1H),3.67(br,4H),2.92(br,4H).HPLC purity:97.3％,Retention time＝12.21min.HRMS(ESI):exact masscalcd for C25H24N7O3[M+H]+,470.1941,found 470.1932.

N- (3- (2- ((4-Morpholphenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 21)

Red solid, yield 87%, mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),10.06(s,1H),8.84(s,1H),8.03(s,1H),7.92(br,1H),7.72(s,1H),7.56(t,J＝7.6Hz,1H),7.27(br,2H),7.12(d,J＝7.2Hz,1H),6.58(br,2H),6.45(dd,J＝16.8,10.4Hz,1H),6.26(d,J＝16.8Hz,1H),5.78(d,J＝10.4Hz,1H),3.71(br,4H),2.94(br,4H).HPLC purity:98.7％,Retention time＝11.71min.HRMS(ESI):exact mass calcd for C₂₅H₂₄N₇O₃[M+H]⁺,470.1941,found 470.1939.

N- (4- (2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO. 22)

Red solid, yield 72%, mp 299.1-299.8 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.51(s,1H),10.06(s,1H),8.83(s,1H),8.03(s,1H),7.89(d,J＝8.4Hz,2H),7.37(d,J＝8.4Hz,2H),7.17(d,J＝6.4Hz,1H),6.56-6.49(m,3H),6.34(d,J＝16.8Hz,1H),5.85(d,J＝10.8Hz,1H),2.94(br,4H),2.37(br,4H),2.20(s,3H).HPLC purity:97.0％,Retention time＝10.02min.HRMS(ESI):exact mass calcd forC₂₆H₂₇N₈O₂[M+H]⁺,483.2257,found 483.2259.

N- (3- (2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO. 23)

Red solid, yield 81%, mp 268.3-269.1 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.45(s,1H),10.06(s,1H),8.84(s,1H),8.04(s,1H),7.93(br,1H),7.73(s,1H),7.56(t,J＝8.0Hz,1H),7.25(br,2H),7.12(d,J＝8.0Hz,1H),6.57(br,2H),6.46(dd,J＝16.8,10.4Hz,1H),6.27(dd,J＝16.8,1.8Hz,1H),5.78(dd,J＝10.4,1.8Hz,1H),2.98(br,4H),2.42(br,4H),2.22(s,3H).HPLC purity:96.5％,Retention time＝9.99min.HRMS(ESI):exact mass calcd for C₂₆H₂₇N₈O₂[M+H]⁺,483.2257,found 483.2259.

N- (3- (7-oxo-2- ((4- (piperidin-1-yl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 24)

Red solid, yield 75%, mp 279.3-280.2 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.44(s,1H),10.03(s,1H),8.83(s,1H),8.02(s,1H),7.94(br,1H),7.73(s,1H),7.55(t,J＝8.0Hz,1H),7.24(br,2H),7.11(d,J＝8.0Hz,1H),6.57(br,2H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),5.77(dd,J＝10.2,1.8Hz,1H),2.95(br,4H),1.57(br,4H),1.49(br,2H).HPLC purity:95.3％,Retention time＝15.12min.HRMS(ESI):exact mass calcd for C₂₆H₂₆N₇O₂[M+H]⁺,468.2148,found 468.2146.

N- (3- (7-oxo-2- ((4- (pyrrolidin-1-yl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 25)

Red solid, 77% yield, mp 279.5-280.1 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.40(s,1H),9.92(s,1H),8.79(s,1H),7.99(s,1H),7.90(br,1H),7.74(br,1H),7.54(t,J＝8.0Hz,1H),7.20(br,2H),7.10(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),6.20(br,2H),5.77(dd,J＝10.2,1.8Hz,1H),3.10(br,4H),1.91(br,4H).HPLC purity:99.3％,Retention time＝15.12min.HRMS(ESI):exact mass calcd for C₂₅H₂₄N₇O₂[M+H]⁺,454.1991,found454.1995.

N- (3- (2- ((4- (diethylamino) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 26)

Red solid, yield 72%, mp 273.5-274.5 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),9.92(s,1H),8.80(s,1H),8.00(s,1H),7.92(br,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.19(br,2H),7.09(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.32(br,2H),6.27(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H),3.20(br,4H),1.00(t,J＝6.8Hz,6H).HPLC purity:96.2％,Retentiontime＝14.69min.HRMS(ESI):exact mass calcd for C₂₅H₂₆N₇O₂[M+H]⁺,456.2148,found456.2143.

N- (3- (2- ((4-Acetylaminophenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 27)

Yellow solid, yield 78%, mp 295.3-295.8 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),10.16(br,1H),9.78(s,1H),8.87(s,1H),8.06(s,1H),7.82(d,J＝8.0Hz,1H),7.79(s,1H),7.56(t,J＝8.0Hz,1H),7.32(br,2H),7.23(br,2H),7.15(d,J＝8.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H),1.98(s,3H).HPLC purity:95.4％,Retention time＝10.68min.HRMS(ESI):exact mass calcd for C₂₃H₂₀N₇O₃[M+H]⁺,442.1628,found 442.1624.

4- ((8- (3-Acylaminophenyl) -7-oxo-7, 8-dihydropteridin-2-yl) amino) benzamide (SEQ ID NO: 28)

Yellow solid, 76% yield, mp 299.3-299.8 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),10.40(s,1H),8.95(s,1H),8.13(s,1H),7.86(s,1H),7.79(d,J＝8.0Hz,1H),7.71(br,1H),7.61(t,J＝8.0Hz,1H),7.55(d,J＝7.6Hz,2H),7.47(br,2H),7.18(d,J＝7.6Hz,2H),6.44(dd,J＝17.0,10.2Hz,1H),6.25(dd,J＝17.0,1.8Hz,1H),5.76(dd,J＝10.2,1.8Hz,1H).HPLC purity:95.6％,Retention time＝10.00min.HRMS(ESI):exact mass calcd for C₂₂H₁₈N₇O₃[M+H]⁺,428.1471,found428.1476.

N- (4- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 29)

Red solid, yield 71%, mp 265.4-266.2 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),8.80(s,1H),8.42(s,1H),8.03(s,1H),7.85(d,J＝8.6Hz,2H),7.34(d,J＝8.6Hz,2H),7.25(d,J＝8.8Hz,1H),6.54-6.48(m,2H),6.33(dd,J＝17.0,1.6Hz,1H),6.02(br,1H),5.84(dd,J＝10.2,1.6Hz,1H),3.76(s,3H),3.02(br,4H),2.43(br,4H),2.23(s,3H).HPLC purity:97.1％,Retention time＝11.44min.HRMS(ESI):exact mass calcd for C₂₇H₂₉N₈O₃[M+H]⁺,513.2363,found513.2362.

N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO. 30)

Red solid, 77% yield, mp 184.7-185.1 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),8.80(s,1H),8.44(br,1H),8.02(s,1H),7.86(br,1H),7.71(s,1H),7.52(t,J＝8.0Hz,1H),7.30(d,J＝7.6Hz,1H),7.09(d,J＝8.0Hz,1H),6.53(s,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,1.8Hz,1H),6.02(br,1H),5.78(dd,J＝10.2,1.8Hz,1H),3.76(s,3H),3.04(br,4H),2.44(br,4H),2.23(s,3H).HPLC purity:96.2％,Retention time＝10.68min.HRMS(ESI):exact mass calcdfor C₂₇H₂₉N₈O₃[M+H]⁺,513.2363,found 513.2361.

N- (4- (2- ((4-methoxyphenyl) amino) -6-methyl-7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 31)

Yellow solid, yield 78%, mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.44(s,1H),9.90(br,1H),8.77(s,1H),7.87(d,J＝8.8Hz,2H),7.50(d,J＝8.8Hz,2H),7.29(br,2H),6.59(br,2H),6.52(dd,J＝17.0,10.0Hz,1H),6.33(dd,J＝17.0,1.9Hz,1H),5.82(dd,J＝10.0,1.9Hz,1H),3.61(s,3H),2.42(s,3H).HPLC purity:95.9％,Retention time＝13.60min.HRMS(ESI):exact mass calcdfor C₂₃H₂₁N₆O₃[M+H]⁺,429.1675,found 429.1671.

N- (3- (2- ((4-methoxyphenyl) amino) -6-methyl-7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 32)

Yellow solid, 66% yield, mp 285.3-286.0 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),9.93(br,1H),8.78(s,1H),7.83(d,J＝8.0Hz,1H),7.77(s,1H),7.56(t,J＝8.0Hz,1H),7.31(br,2H),7.11(d,J＝8.0Hz,1H),6.58(br,2H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(d,J＝17.0Hz,1H),5.77(d,J＝10.2Hz,1H),3.65(s,3H),2.42(s,3H).HPLC purity:98.6％,Retention time＝13.51min.HRMS(ESI):exact mass calcd for C₂₃H₂₁N₆O₃[M+H]⁺,429.1675,found 429.1675.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((2-methoxy) -4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 33)

Orange-red solid, yield 40%.

¹H NMR(400MHz,DMSO-d₆):δ8.96(s,1H),8.73(s,0.5H),8.72(s,0.5H),7.86(s,0.5H),7.84(s,0.5H),7.32(m,1H),6.57(s,1H),6.43-6.42(m,1H),6.16(ddd,J＝16.8,10.0,2.4Hz,1H),5.72(dd,J＝10.0,2.4Hz,1H),5.65(dd,J＝10.0,2.4Hz,1H),4.08(t,J＝10.2Hz,0.5H),3.88-3.82(m,0.5H),3.76(s,3H),3.67-3.62(m,1H),3.58-3.55(m,1H),3.21-3.17(m,4H),2.81-2.71(m,1H),2.68-2.61(m,4H),2.38(s,1.4H),2.34(s,1.6H),2.04-1.96(m,2H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₂₅H₃₁N₈O₃491.2519; found, 491.2520. HPLC purity 95.7%, retention time 9.43min.

(R) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((2-methoxy) -4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 34)

Orange-red solid, yield 40%.

¹H NMR(400MHz,DMSO-d₆):δ8.94(s,1H),8.72-8.71(m,1H),7.86-7.84(m,1H),7.33(t,J＝8.4Hz,1H),6.56(s,1H),6.42-6.40(m,1H),6.16(ddd,J＝16.8,10.4,2.4Hz,1H),5.70(dd,J＝10.4,2.4Hz,1H),5.65(dd,J＝10.4,2.4Hz,1H),4.10(t,J＝10.2Hz,0.5H),3.88-3.83(m,0.6H),3.76(s,3H),3.67-3.62(m,1H),3.58-3.55(m,1H),3.15-3.10(m,4H),2.82-2.64(m,1H),2.45(br,4H),2.23(s,3H),2.06-1.96(m,1H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₅H₃₁N₈O₃,491.2519；found,491.2473.

(S) -8- (1-acryloyl-3-piperidinyl) -2- ((2-methoxy) -4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 35)

Orange-red solid, yield 38%.

¹H NMR(400MHz,DMSO-d₆):δ9.14(br,1H),8.71(s,1H),7.83(s,1H),7.22-7.21(m,1H),6.80-6.62(m,2H),6.48-6.47(m,1H),6.15-6.08(m,1H),5.71-5.60(m,1H),4.82-4.76(m,0.6H),4.34-4.24(m,1H),3.95-3.92(m,1H),3.74(s,3H),3.14(br,4H),2.46(br,4H),1.64(br,2H),1.34-1.30(m,1H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₆H₃₃N₈O₃,505.2676；found,505.2676.

N- (3- (2- ((3-methoxy-4- (methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 36)

Red solid, yield 55%.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),10.00(br,1H),8.86(s,1H),8.04(s,1H),7.87(d,J＝8.0Hz,1H),7.73(s,1H),7.54(t,J＝8.0Hz,1H),7.11(d,J＝8.0Hz,1H),7.04-6.95(m,2H),6.48-6.42(m,2H),6.27(dd,J＝17.2,2.0Hz,1H),5.77(dd,J＝10.0,2.0Hz,1H),3.55(s,3H),2.83(br,4H),2.43(br,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₇H₂₉N₈O₃,513.2363；found,513.2362.

N- (3- (2- ((3-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo) -8(7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 37)

Yellow solid, yield 63%.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),10.04(s,1H),8.86(s,1H),8.04(s,1H),7.87(d,J＝8.0Hz,1H),7.76(s,1H),7.55(t,J＝8.0Hz,1H),7.22(s,1H),7.13-7.10(m,2H),6.70-6.69(m,1H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,2.0Hz,1H),5.76(dd,J＝10.2,2.0Hz,1H),2.70(br,4H),2.44(br,4H),2.23(s,3H),1.97(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₇H₂₉N₈O₂,497.2413；found,497.2428.

N- (3- (2- ((2-methoxy-5-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 38)

Red solid, 50% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.38(s,1H),8.82(s,1H),8.37(s,1H),8.04(s,1H),7.82(d,J＝8.0Hz,1H),7.74(s,1H),7.53(t,J＝8.0Hz,1H),7.29(s,1H),7.10(d,J＝8.8Hz,1H),6.62(s,1H),6.45(dd,J＝16.8,2.0Hz,1H),6.25(dd,J＝16.8,2.0Hz,1H),5.77-5.76(m,1H),3.78(s,1H),2.76(br,4H),2.46(br,4H),2.24(s,3H),1.85(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₈H₃₁N₈O₃,527.2519；found,527.2518.

N- (3- (2- ((4- (4-acetyl-1-piperazinyl) -2-methoxyphenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 39)

Red solid, 50% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.36(s,1H),8.80(s,1H),8.44(s,1H),8.03(s,1H),7.86(d,J＝8.0Hz,1H),7.69(t,J＝2.0Hz,1H),7.52(t,J＝8.0Hz,1H),7.32(d,J＝8.8Hz,1H),7.09(d,J＝8.0Hz,1H),6.57(d,J＝2.4Hz,1H),6.45(dd,J＝17.0,2.0Hz,1H),6.27(dd,J＝17.0,10.0Hz,1H),6.09(br,1H),5.77(dd,J＝10.0,2.0Hz,1H),3.77(s,1H),3.58-3.54(m,4H),3.06-3.00(m,4H),2.05(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₂₈H₂₉N₈O₄,541.2312；found,541.2312.

N- (3- (2- ((4- (2- (4-methyl-1-piperazinyl) ethyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 40)

Yellow solid, yield 50%.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),10.13(s,1H),8.88(s,1H),8.07(s,1H),7.88(d,J＝8.0Hz,1H),7.77(s,1H),7.57(t,J＝8.0Hz,1H),7.31-7.30(m,2H),7.14(d,J＝8.0Hz,1H),6.86(br,2H),6.45(dd,J＝16.8,10.0Hz,1H),6.26(dd,J＝16.8,2.0Hz,1H),5.77(dd,J＝10.0,2.0Hz,1H),2.56(t,J＝7.6Hz,2H),2.39-2.31(m,9H),2.15(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₈H₃₁N₈O₂,511.2570；found,511.2571.

N- (3- (2- ((2- (methoxyethoxy) -4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 41)

Orange-red solid, yield 61%.

¹H NMR(400MHz,CDCl₃):δ8.78(s,1H),8.65-8.62(m,1H),8.24(s,1H),8.07(s,1H),7.85(s,1H),7.41(br,3H),6.93(d,J＝5.6Hz,1H),6.44(s,1H),6.33(d,J＝17.0Hz,1H),6.10(dd,J＝17.0,10.4Hz,1H),5.78(d,J＝10.4Hz,1H),4.10(br,2H),3.69(br,2H),3.43(s,3H),3.08(br,4H),2.54(br,4H),2.34(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₂₉H₃₃N₈O₄,557.2625；found,557.2621.

N- (3- (2- ((4- (4- (2-hydroxyethyl) -1-piperazinyl) -2-methoxyphenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 43)

Orange solid, yield 53%.

¹H NMR(400MHz,DMSO-d₆):δ10.39(s,1H),8.79(s,1H),8.43(s,1H),8.02(s,1H),7.85(d,J＝7.6Hz,1H),7.71(s,1H),7.52(t,J＝8.0Hz,1H),7.30(d,J＝7.6Hz,1H),7.09(d,J＝8.0Hz,1H),6.52(s,1H),6.45(dd,J＝16.8,10.0Hz,1H),6.27(dd,J＝16.8,2.0Hz,1H),6.02(br,1H),5.78(dd,J＝10.0,2.0Hz,1H),4.43(t,J＝5.2Hz,1H),3.76(s,3H),3.56-3.52(m,2H),3.04(br,4H),2.53(t,J＝4.8Hz,4H),2.44(t,J＝6.0Hz,1H).HRMS(ESI)(m/z):[M+H]⁺calcdfor C₂₉H₃₃N₈O₃,543.2468；found,543.2466.

N- (3- (2- ((4- (2- (4-methyl-1-piperazinyl) ethyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 44)

Red solid, 50% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),8.80(s,1H),8.43(s,1H),8.03(s,1H),7.88-7.87(m,1H),7.70(s,1H),7.52(t,J＝8.0Hz,1H),7.29(d,J＝8.0Hz,1H),7.09(d,J＝8.0Hz,1H),6.52(s,1H),6.46(dd,J＝17.2,10.0Hz,1H),6.27(dd,J＝17.2,2.0Hz,1H),6.04(br,1H),5.78(dd,J＝10.0,2.0Hz,1H),3.76(s,3H),3.60-3.58(m,2H),2.58(t,J＝10.8Hz,2H),2.20-2.18(m,7H),1.82-1.79(d,J＝12Hz,2H),1.48-1.40(m,2H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₉H₃₃N₈O₃,541.2676；found,541.2674.

N- (3- (6-isopropyl-2- ((4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 45)

163mg of red solid, yield 75%.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),9.85(br,1H),8.79(s,1H),7.91(d,J＝6.4Hz,1H),7.71(s,1H),7.55(t,J＝8.0Hz,1H),7.25(s,2H),7.11(d,J＝8.0Hz,1H),6.59-6.58(m,2H),6.45(dd,J＝17.0,10.2Hz,1H),6.26(dd,J＝17.0,2.0Hz,1H),5.78(dd,J＝10.2,2.0Hz,1H),3.45-3.38(m,1H),2.97(br,4H),2.42(br,4H),2.21(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₉H₃₃N₈O₂,525.2726；found,525.2728.

N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO. 46)

Red solid, yield 75%.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),8.87(s,1H),8.41(s,1H),8.21-8.19(m,2H),7.89-7.88(m,1H),7.75(t,J＝2.0Hz,1H),7.54(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.35(d,J＝8.8Hz,1H),7.17-7.14(m,1H),6.54(d,J＝2.0Hz,1H),6.46(dd,J＝17.0,10.2Hz,1H),6.27(dd,J＝17.0,2.0Hz,1H),6.05(br,1H),5.78(dd,J＝10.2,2.0Hz,1H),3.78(s,3H),3.05(br,4H),2.45(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₃₃H₃₃N₈O₃,589.2676；found,589.2676.

8- (1-acryloyl-4-piperidinyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 47)

Orange solid, yield 53%.

¹H NMR(400MHz,DMSO-d₆):δ8.83(s,1H),8.71(s,1H),7.83(s,1H),7.53(br,1H),6.85(dd,J＝17.2,10.4Hz,1H),6.61(d,J＝2.4Hz,1H),6.47(d,J＝8.0Hz,1H),6.13(dd,J＝17.2,2.4Hz,1H),5.70(dd,J＝10.4,2.4Hz,1H),5.29(br,1H),4.60(d,J＝10.4Hz,1H),4.20(d,J＝12.8Hz,1H),3.80(s,3H),3.13(t,J＝4.8Hz,4H),2.59-2.53(m,3H),2.45(t,J＝4.8Hz,4H),2.22(s,3H),1.67(d,J＝10.0Hz,2H),1.23(s,1H).

(R) -8- ((1-acryloyl-3-piperidinyl) methyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (Ser. No. 48)

Orange solid, yield 61%.

¹H NMR(400MHz,DMSO-d₆):δ8.92(s,1H),8.74(s,1H),7.91-7.89(m,1H),7.50-7.44(m,1H),6.75(dd,J＝16.8,10.4Hz,1H),6.64(s,1H),6.56-6.51(m,1H),6.02(d,J＝16.8Hz,1H),5.64-5.54(m,1H),4.13-4.10(m,1H),3.94-3.83(m,3H),3.76(s,3H),3.16(br,4H),3.03-2.98(m,1H),2.84-2.74(m,1H),2.47(t,J＝4.8Hz,4H),1.92-1.91(m,1H),1.64-1.57(m,2H),1.30-1.23(m,3H).

(S) -8- ((1-acryloyl-3-piperidinyl) methyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 49)

Orange solid, yield 54%.

¹H NMR(400MHz,DMSO-d₆):δ8.92(s,1H),8.74(s,1H),7.91-7.89(m,1H),7.49-7.43(m,1H),6.75(dd,J＝16.4,10.4Hz,1H),6.64(s,1H),6.56-6.51(m,1H),6.02(d,J＝16.4Hz,1H),5.64-5.54(m,1H),4.13-4.08(m,1H),3.94-3.86(m,3H),3.76(s,3H),3.16(br,4H),3.03-2.97(m,1H),2.84-2.74(m,1H),2.47(t,J＝4.8Hz,4H),1.92(br,1H),1.64-1.57(m,2H),1.30-1.20(m,3H).

N- (3- (2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxyphenyl) amino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 50)

Red solid, yield 61%.

¹H NMR(400MHz,DMSO-d₆):δ10.37(s,1H),8.76(s,1H),8.41(br,1H),7.99(s,1H),7.84-7.83(m,1H),7.69(t,J＝2.0Hz,1H),7.51(t,J＝8.0Hz,1H),7.26-7.19(m,1H),7.09(d,J＝8.0Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.29-6.24(m,2H),5.77(dd,J＝10.4,2.0Hz,1H),3.74(s,3H),2.84(s,3H),2.35-2.33(m,2H),2.19(s,6H).

N- (3- (2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxyphenyl) amino) -6-isopropyl-7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 51)

Orange solid, yield 62%.

¹H NMR(400MHz,DMSO-d₆):δ10.37(s,1H),8.72(s,1H),8.21(s,1H),7.84(d,J＝8.0Hz,1H),7.69(s,1H),7.51(t,J＝8.0Hz,1H),7.26(d,J＝7.6Hz,1H),7.09(d,J＝8.4Hz,1H),6.45(dd,J＝16.8,10.4Hz,1H),6.29-6.24(m,2H),5.87(br,1H),5.76(dd,J＝10.4,2.0Hz,1H),3.75(s,3H),3.43-3.38(m,1H),2.84(s,3H),2.34(t,J＝6.8Hz,2H),2.19(s,6H),1.24(d,J＝6.8Hz,6H).

N- (3- (2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) -2-methoxyphenyl) amino) -7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 52)

Red solid, yield 64%.

¹H NMR(400MHz,DMSO-d₆):δ10.39(s,1H),8.83(s,1H),8.38(br,1H),8.21-8.18(m,2H),7.87(d,J＝7.2Hz,1H),7.75(s,1H),7.53(t,J＝8.0Hz,1H),7.49-7.47(m,3H),7.28(br,1H),7.15(d,J＝8.4Hz,1H),6.46(dd,J＝16.8,10.0Hz,1H),6.29-6.25(m,2H),5.83(br,1H),5.76(dd,J＝10.0,2.0Hz,1H),3.76(s,3H),3.35(br,2H),2.85(s,3H),2.35(t,J＝5.6Hz,2H),2.20(s,6H).

(R) -8- ((1-acryloyl-3-piperidinyl) methyl) -6-isopropyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 53)

Yellow solid, yield 55%.

¹H NMR(400MHz,DMSO-d₆):δ8.70(br,2H),7.57-7.52(m,1H),6.76(dd,J＝16.4,10.4Hz,1H),6.64(s,1H),6.57-6.51(m,1H),6.05-5.98(m,1H),5.64-5.53(m,1H),4.15-3.96(m,3H),3.90-3.87(m,1H),3.78(s,3H),3.16(br,4H),3.04-2.75(m,2H),2.47(t,J＝3.6Hz,4H),2.24(s,3H),1.94(br,1H),1.66-1.61(m,2H),1.20(d,J＝6.8Hz,6H).

(R) -8- (1-acryloyl-3-piperidinyl) -2- ((2-methoxy) -4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 54)

Orange solid, yield 46%.

¹H NMR(400MHz,DMSO-d₆):δ9.17(s,1H),8.71(s,1H),7.83(s,1H),7.20(br,1H),6.81(br,1H),6.68-6.62(m,2H),6.47(br,1H),6.14-6.08(m,1H),5.72-5.61(m,1H),4.78-4.72(m,1H),4.35-4.24(m,1H),3.95-3.82(m,1H),3.73(s,3H),3.14(br,4H),2.46(br,4H),2.23(s,3H),1.62(br,2H),1.34-1.26(m,1H).

N- (2- ((2- (dimethylamino) ethyl) (methyl) amino) -5- ((6-isopropyl-8-methyl) -7-oxo-7, 8-dihydro-2-pteridinyl) amino) -4-methoxyphenyl) acrylamide (SEQ ID NO: 55)

Yellow solid, 43% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.08(s,1H),8.96(s,1H),8.73(s,1H),8.64(s,1H),7.02(s,1H),6.42(dd,J＝16.4,10.0Hz,1H),6.24(d,J＝16.4Hz,1H),5.75(d,J＝10.4Hz,1H),3.85(s,3H),3.59(s,3H),3.43-3.36(m,1H),2.89(br,2H),2.71(s,3H),2.35(br,2H),2.23(s,6H),1.20(d,J＝6.8Hz,6H).

(S) -8- (1-acryloyl-3-pyrrolidinyl) -6-isopropyl-2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 56)

Yellow solid, yield 81%.

¹H NMR(400MHz,DMSO-d₆):δ9.90(s,1H),8.77(s,0.6H),8.76(s,0.4H),7.49-7.41(m,2H),6.93(t,J＝7.2Hz,1H),6.73-6.47(m,1H),6.23-6.15(m,1H),6.03-5.94(m,1H),5.76-5.65(m,1H),4.22(t,J＝8.8Hz,0.6H),4.02-3.86(m,1.7H),3.83-3.65(m,1.7H),3.47-3.40(m,1H),2.92-2.85(m,1H),2.78(br,4H),2.46(br,4H),2.23(s,3H),2.20(s,3H),2.14-2.07(m,1H),1.20(d,J＝6.4Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₂₈H₃₇N₈O₂[M+H]⁺517.3039；found,517.3038.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) -pteridinone (SEQ ID NO: 57)

Orange solid, yield 77%.

¹H NMR(400MHz,DMSO-d₆):δ10.09(s,1H),8.88(s,1H),8.13(br,2H),7.52-7.49(m,5H),6.98-6.96(m,1H),6.74-6.48(m,1H),6.23-6.15(m,1H),6.13-6.02(m,1H),5.73-5.65(m,1H),4.28-4.24(m,0.5H),4.12-3.90(m,1.7H),3.85-3.67(m,1.8H),3.52-3.44(m,1H),2.94-2.89(m,1H),2.82(br,4H),2.28(s,3H),2.22(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₅N₈O₂[M+H]⁺551.2883；found,551.2880.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -6-isopropyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 58)

Orange solid, yield 54%.

¹H NMR(400MHz,DMSO-d₆):δ8.71(s,1H),8.69(s,0.5H),8.68(s,0.5H),7.39(t,J＝7.6Hz,1H),6.57(s,1H),6.41(d,J＝8.8Hz,1H),6.17(dd,J＝16.4Hz,10.8Hz,1H),5.86-5.80(m,1H),5.72-5.63(m,1H),4.12(t,J＝8.8Hz,0.6H),3.91-3.86(m,0.6H),3.77(s,3H),3.69-3.64(m,1H),3.59(br,1H),3.40-3.38(m,0.8H),3.12(br,4H),2.81-2.70(m,1H),2.45(br,4H),2.23(s,3H),2.09-1.98(m,1H),1.18(d,J＝6.4Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for 533.2989；found,533.2994.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) -pteridinone (SEQ ID NO: 59)

Red solid, yield 63%.

¹H NMR(400MHz,DMSO-d₆):δ8.90(s,1H),8.79(s,1H),8.10(br,2H),7.46(br,3H),7.41-3.39(m,1H),6.58-6.42(m,3H),6.17(dd,J＝16.4Hz,10.4Hz,1H),5.98-5.82(m,1H),5.72-5.64(m,1H),4.16(t,J＝8.8Hz,0.6H),3.95-3.90(m,0.6H),3.78(s,3H),3.71(t,J＝9.6Hz,0.8H),3.60(br,1H),3.40-3.34(m,1H),3.13(br,4H),2.83-2.71(m,1H),2.45(br,4H),2.23(s,3H),2.12-2.02(m,1H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₅N₈O₃[M+H]⁺567.2832；found,567.2831.

(S, E) -8- (1- (4- (dimethylamino) -2-butenoyl) -3-pyrrolidinyl) -2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) -pteridinone (SEQ ID NO: 61)

Orange solid, yield 82%.

¹H NMR(400MHz,DMSO-d₆):δ10.10(s,1H),8.88(s,1H),8.12(s,2H),7.49(br,5H),6.97(d,J＝8.4Hz,1H),6.72-6.61(m,1H),6.54-6.31(m,1H),6.10-6.00(m,1H),4.28-4.24(m,0.6H),4.05-4.00(m,0.7H),3.93-3.86(m,1.5H),3.81-3.76(m,1.5H),3.73-3.67(m,1H),3.55(br,4H),3.18(d,J＝6.8Hz,2H),3.08(d,J＝5.2Hz,1H),2.83(br,4H),2.57(s,3H),2.32(s,1.5H),2.30(s,1.5H),2.57(s,3H),2.22(s,3H),2.17(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₄₂N₉O₂[M+H]⁺608.3461；found,608.3466.

(E) -8- (1- (4- (dimethylamino) -2-butenoyl) -4-piperidinyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) -pteridinone (SEQ ID NO: 62)

Red solid, 71% yield, mp 157.3-157.6 ℃.

¹H NMR(400MHz,DMSO-d₆):δ8.86(s,1H),8.72(s,1H),7.83(s,1H),7.54(br,1H),6.64-6.62(m,3H),6.47(d,J＝7.6Hz,1H),5.27(br,1H),4.61-4.58(m,1H),4.21-4.18(m,1H),3.79(s,3H),3.14(br,4H),3.05-3.04(br,2H),2.57(br,4H),2.46(t,J＝4.0Hz,4H),2.23(s,3H),2.16(s,6H),1.67(br,2H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₉H₄₀N₉O₃,562.3254；found,562.3259.

N- (3- (6-isopropyl-2- ((3-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 64)

Yellow solid, yield 69%. mp 249.0-249.5 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),9.83(br,1H),8.82(s,1H),7.87(d,J＝8.0Hz,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),7.04(br,1H),6.96(br,1H),6.49-6.42(m,2H),6.26(dd,J＝16.8Hz,2.0Hz,1H),5.77(dd,J＝10.0Hz,2.0Hz,1H),3.55(s,3H),3.44-3.39(m,1H),2.82(br,4H),2.42(br,4H),2.21(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₀H₃₅N₈O₃,555.2832；found,555.2838.

N- (3- (2- ((3-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-6- (trifluoromethyl) -8(7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 65)

Brown solid, yield 59%, mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.44(s,1H),10.40(s,1H),9.00(s,1H),7.88(d,J＝7.2Hz,1H),7.81(s,1H),7.56(t,J＝8.0Hz,1H),7.14(d,J＝8.0Hz,1H),7.01(d,J＝7.2Hz,1H),6.95(s,1H),6.49-6.42(m,2H),6.27(d,J＝16.4Hz,1H),5.77(d,J＝10.4Hz,1H),3.53(s,3H),2.83(br,4H),2.41(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₂₈H₂₈N₈O₃F₃,581.2236；found,581.2241.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 66)

Yellow solid, yield 45%, mp 215.5-215.7 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.08(s,1H),8.81(d,J＝3.2Hz,1H),7.89(d,J＝6.8Hz,1H),7.47-7,42(m,2H),6.93(t,J＝6.8Hz,1H),6.72-6.46(m,1H),6.22-6.6.14(m,1H),5.98-5.88(m,1H),5.75-5.64(m,1H),4.20(t,J＝8.8,1H),4.00-3.64(m,4H),3.47-3.40(m,1H),2.78(br,4H),2.45(s,3H),2.23(s,3H),2.20(br,4H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₂₅H₃₁N₈O₂,475.2570；found,475.2550.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((3-methoxy-4 (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 67)

Orange solid, 40% yield, mp 208.3-208.8 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.10(s,1H),8.88(d,J＝3.2,1H),8.13-8.12(m,2H),7.48(d,J＝2.4Hz,1H),7.47(s,1H),7.34-7.27(m,2H),6.81(t,J＝8.4Hz,1H),6.74-6.48(m,1H),6.23-6.15(m,1H),6.12-6.04(m,1H),5.76-5.65(m,1H),4.25(t,J＝9.2,1H),4.13-3.76(m,3H),3.76(s,3H),3.74-3.67(m,1H),3.51-3.44(m,1H),2.91(br,4H),2.44(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₅N₈O₃,567.2832；found,567.2835.

(S, E) -8- (1- (4- (dimethylamino) -2-butenoyl) 3-pyrrolidinyl) -2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-isopropyl-7 (8H) pteridinone (SEQ ID NO: 68)

Red solid, yield 62%, mp 159.9-160.7 ℃.

¹H NMR(400MHz,DMSO-d₆):δ9.94(s,1H),8.77(d,J＝3.6Hz,1H),7.49-7.40(m,2H),6.94(d,J＝8.4,1H),6.72-6.61(m,1H),6.53(d,J＝15.2Hz,0.5H),6.34(d,J＝15.2Hz,0.5H),6.04-5.91(m,1H),4.22(t,J＝8.4Hz,1H),4.00-3.64(m,4H),3.40-3.35(m,2H),3.22(t,J＝6.4Hz,1H),3.10(d,J＝6.0Hz,1H),2.82(br,4H),2.61(br,4H),2.34(d,11.2Hz,3H),2.28(s,3H),2.20(br,4H),2.19(s,3H),1.20(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₄₄N₉O₂,574.3618；found,574.3584.

N- (3- (6-isopropyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 69)

Orange solid, yield 65%, mp 242.9-243.9 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),8.75(s,1H),8.26(s,1H),7.87(d,J＝7.2Hz,1H),7.71(s,1H),7.53(t,J＝8.0Hz,1H),7.34(d,J＝8.8Hz,1H),7.10(d,J＝8.8Hz,1H),6.53(d,J＝2.0Hz,1H),6.46(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,2.0Hz,1H),6.12-6.00(m,1H),5.78(dd,J＝10.0Hz,2.0Hz),3.76(s,3H),3.44-3.41(m,1H),3.04(br,4H),2.43(t,J＝4.8Hz,4H),2.22(s,3H),1.25(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₀H₃₅N₈O₃,555.2832；found,555.2821.

N- (3- (2- ((2-methoxy-5-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-6-phenyl-8 (7H) pteridinonyl) phenyl) acrylamide (SEQ ID NO: 70)

Yellow solid, yield 62%, mp 292.9-293.4 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.41(s,1H),8.90(s,1H),8.36(br,1H),8.21-8.18(m,2H),7.84(d,J＝8.0Hz,1H),7.80(s,1H),7.55(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.33(s,1H),7.16(d,J＝7.6Hz,1H),6.63(s,1H),6.45(dd,J＝16.8Hz,10.0Hz,1H),6.26(dd,J＝16.8Hz,1.6Hz),5.77(dd,J＝10.0Hz,1.6Hz,1H),3.79(s,3H),2.77(br,4H),2.48(br,4H),2.25(s,3H),1.85(br,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₃₅N₈O₃,603.2832；found,603.2834.

N- (3- (6-isopropyl-2- ((2-methoxy-5-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 71)

Orange solid, 60% yield, mp 227.5-228.5 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.40(s,1H),8.79(s,1H),8.22(s,1H),7.82(d,J＝8.0Hz,1H),7.73(s,1H),7.53(t,J＝8.0Hz,1H),7.31(s,1H),7.11(d,J＝7.6Hz,1H),6.62(s,1H),6.44(dd,J＝16.8Hz,10.0Hz,1H).6.25(dd,J＝16.8Hz,0.8Hz,1H),5.75(d,J＝10.0Hz,1H),3.78(s,3H),3.43-3.39(m,1H),2.77(br,4H),2.26(s,3H),1.85(br,3H),1.24(d,J＝6.8Hz,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₇N₈O₃,569.2989；found,569.2989.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) -3-methylphenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 72)

Red solid, 40% yield, mp 256.1-256.4 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.08(s,1H),8.88(d,J＝3.2Hz,1H),8.13-8.12(m,2H),7.52-7.45(m,5H),7.02(dd,J＝8.4Hz,5.6Hz,1H),6.74-6.49(m,1H),6.23-6.15(m,1H),6.12-6.02(m,1H),5.75-5.65(m,1H),4.28-4.23(m,1H),4.15-4.12(m,1H),3.94-3.68(m,3H),3.52-3.44(m,1H),2.90(t,J＝6.8Hz,2H),2.61(s,3H),2.35(t,J＝6.8Hz,2H),2.22(s,3H),2.13(br,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₇N₈O₂,553.3039；found,553.3031.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -2- ((4- ((2- (dimethylamino) ethyl) (methyl) amino) -3-methoxyphenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 73)

Dark red solid, yield 49%, mp 260.0-260.2 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.09(s,1H),8.88(d,J＝2.8Hz,1H),8.131(br,2H),7.49-7.48(m,3H),7.34-7.25(m,2H),6.81(t,J＝7.2Hz,1H),6.76-6.49(m,1H),6.23-6.05(m,2H),5.75-5.65(m,1H),4.25(t,J＝8.4Hz,1H).4.06-3.82(m,3H),3.77(s,3H),3.74-3.67(m,1H),3.05(t,J＝7.2Hz,2H),2.94-2.79(m,1H),2.68(s,3H),2.36(t,J＝7.2Hz,2H),2.13(s,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₃₇N₈O₃,569.2989；found,569.2988.

N- (3- (2- ((3-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-6-phenyl-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 74)

Red solid, yield 81%, mp 264.7-265.5 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),10.01(s,1H),8.93(s,1H),8.22-8.19(m,2H),7.90(d,J＝7.6Hz,1H),7.79(s,1H),7.56(t,J＝8.0Hz,1H),7.49(t,J＝3.2Hz,1H),7.17(d,J＝8.4Hz,1H),7.07(br,1H),6.97(br,1H),6.46(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),3.56(s,3H),2.83(br,4H),2.41(br,4H),2.21(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₃N₈O₃,589.2676；found,589.2642.

N- (3- (6-cyclohexyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 75)

Yellow solid, yield 82%, mp 268.9-269.4 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.40(s,1H),8.73(s,1H),8.24(s,1H),7.85(d,J＝8.0Hz,1H),7.69(t,J＝1.6Hz,1H),7.52(t,J＝8.0Hz,1H),7.33(d,J＝9.2Hz,1H),7.10-7.08(m,1H),6.53(d,J＝2.4Hz,1H),6.45(dd,J＝16.8Hz,10.0Hz,1H),6.26(dd,J＝16.8Hz,2.0Hz,1H),6.06(br,1H),5.77(dd,J＝10.0Hz,2.0Hz,1H),3.76(s,3H),3.03(br,4H),2.43(t,J＝4.8Hz,4H),2.22(s,3H),3.41(d,J＝12.0Hz,2H),1.82(d,J＝12.0Hz,2H),1.72(d,J＝12.0Hz,2H),1.53-1.23(m,6H)。HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₉N₈O₃,595.3145；found,595.3141.

(S) -8- ((1-acryloyl-3-piperidyl) methyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 76)

Red solid, yield 49%, mp 208.3-209.3 ℃.

¹H NMR(400MHz,DMSO-d₆):δ8.88(s,1H),8.81(s,1H),8.18-8.16(m,2H),7.59-7.52(m,1H),7.47(t,J＝3.2Hz,3H),6.75(dd,J＝16.4Hz,10.0,1H),6.64(s,1H),6.55(dd,J＝17.2Hz,8.8Hz,1H),6.03(d,J＝16.4Hz,1H),5.58(dd,J＝32.8Hz,10.0Hz,1H),4.19-4.06(m,3H),3.89-3.83(m,1H),3.79(s,3H),3.17(br,4H),3.05-2.55(m,2H),2.48(t,J＝4.8Hz,4H),2.24(s,3H),2.00(br,1H),1.66(br,2H),1.26-1.23(m,2H)。HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₉N₈O₃,595.3145；found,595.3157.

(R) -8- (1-acryloyl-3-piperidinyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 77)

Red solid, yield 38%, mp 134.9-135.1 ℃.

¹H NMR(400MHz,DMSO-d₆):δ9.17(s,1H),8.79(s,1H),8.11-8.09(m,2H),7.77-7.46(m,3H),7.24(br,1H),6.84-6.63(m,2H),6.49(s,1H),6.06-6.19(m,1H),5.67(dd,J＝37,2Hz,9.6Hz,1H),5.00-4.27(m,3H),3.75(s,3H),3.14(s,4H),2.49(s,4H),2.24(s,3H),1.69(br,2H),1.35(br,1H),1.23(s,1H)。HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₂H₃₇N₈O₃,581.2989；found,581.2972.

(R) -8- (1-acryloyl-3-piperidinyl) -6-cyclohexyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 78)

Yellow solid, yield 53%, mp 94.2-94.7 ℃.

¹H NMR(400MHz,DMSO-d₆):δ8.96(s,1H),8.65(s,1H),7.23(br,1H),6.83-6.61(m,2H),6.47(s,1H),6.15-6.08(m,1H),5.66(dd,J＝36Hz,9.6Hz,1H),4.84(br,1H),4.29(br,1H),3.95(br,1H),3.72(s,3H),3.13(br,4H),2.46(br,4H),2.23(s,3H),1.81-1.63(m,7H),1.44-1.23(m,9H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₂H₄₃N₈O₃,587.3458；found587.3458.

N- (3- (2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-6-phenyl-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 79)

Yellow solid, yield 68%, mp 265.0-265.2 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),10.04(s,1H),8.92(s,1H),8.22-8.19(m,2H),7.89(d,J＝8.0Hz,1H),7.81(s,1H),7.57(t,J＝8.0Hz,1H),7.50-7.49(m,3H),7.25(s,1H),7.19(s,1H),7.17(s,1H),6.71(br,1H),6.46(dd,J＝16.8Hz,10.4Hz,1H),6.26(dd,J＝16.8Hz,1.6Hz,1H),5.77(dd,J＝10.4Hz,1.6Hz,1H),2.7(br,4H),2.44(br,4H),2.23(s,3H),1.98(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₃N₈O₂,573.2726；found,573.2729.

N- (3- (2- ((2-methoxy-4- (4- (4-methyl-1-piperazinyl) piperidinyl) phenyl) amino) -7-oxo-6-phenyl-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 80)

Red solid, yield 40%, mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.44(s,1H),8.86(s,1H),8.41(s,1H),8.20-8.19(m,2H),7.90(br,1H),7.75(s,1H),7.54(t,J＝8.0Hz,1H),7.50-7.48(m,3H),7.33(d,J＝6.4Hz,1H),7.15(d,J＝8.0Hz,1H),6,52(br,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.04(br,1H),5.78(dd,J＝10.0Hz,1.6Hz),3.77(s,3H),3.61(d,J＝8.4Hz,2H),2.60-2.54(m,5H),2.39-2.30(m,5H),1,81(d,J＝11.6Hz,2H),1.51-1.43(m,2H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₈H₄₂N₉O₃,672.3411；found,672.3407.

8- (1-acryloyl-3-pyrrolidinyl) -2- ((3-methyl-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 81)

Orange solid, yield 65%.

¹H NMR(400MHz,DMSO-d₆):δ10.08(s,1H),8.87(d,J＝2.8Hz,1H),8.13-8.11(m,2H),7.51-7.46(m,5H),6.97-6.93(m,1H),6.70(dd,J＝16.8Hz,10.4,1H),6.51(dd,J＝16.8Hz,10.4Hz,1H),6.23-6.15(m,1H),6.11-6.01(m,1H),5.75-5.65(m,1H),4.25(t,J＝8.8Hz,1H),4.06-3.68(m,3H),3.52-3.45(m,1H),2.95-2.86(m,1H),2.79(br,4H),2.46(br,4H),2.23(s,3H),2.21(s,3H),2.17-2.12(m,1H).HRMS(ESI)(m/z):[M+H]⁺calcd forC₃₁H₃₅N₈O₂,551.2883；found,551.2883.

(S) -8- (1-acryloyl-3-pyrrolidinyl) -6-cyclohexyl-2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7(8H) pteridinone (SEQ ID NO: 82)

Orange solid, yield 49%, mp 117.9-118.2 ℃.

¹H NMR(400MHz,DMSO-d₆):δ8.71(s,1H),8.66(d,J＝3.6Hz,1H),7.39(t,J＝8.0Hz,1H),6.63-6.59(m,0.5H),6.57(s,1H),6.49-6.45(m,0.5H),6.41(d,J＝9.2Hz,1H),6.20-6.13(m,1H),5.82(br,1H),5.73-5.63(m,1H),4.11(t,J＝9.2Hz,0.5H),3.87(dd,J＝12.0Hz,8.8Hz,0.5H),3.69-3.58(m,2H),3.18-3.04(m,5H),2.80-2.65(m,1H),2.45(br,4H),2.23(s,3H),2.09(br,0.5H),1.99(br,0.5H),1.81(t,J＝11.2Hz,4H),1.70(d,J＝11.2Hz,1H),1.45-1.16(m,6H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₁H₄₁N₈O₃,573.3302；found,573.3304.

8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6-phenyl-7 (8H) pteridinone (SEQ ID NO: 83)

Orange solid, yield 84%, mp 235.5-235.7 ℃.

¹H NMR(400MHz,DMSO-d₆):δ8.84(s,1H),8.39(br,1H),8.19-8.17(m,2H),7.49-7.47(m,4H),7.21(t,J＝8.0Hz,1H),6.72(d,J＝7.2Hz,1H),6.571-6.56(m,2H),6.52(d,J＝8.0Hz,1H),6.19(br,1H),5.34(s,2H),3.79(s,3H),3.08(br,4H),2.45(t,J＝4.4Hz,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₀H₃₁N₈O₂,535.2570；found,535.2569.

N- (3- (6- (4-fluorophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 84)

Orange solid, 73% yield, mp 262.3-262.7 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),8.87(s,1H),8.44(s,1H),8.29(dd,J＝8.4Hz,6.0Hz,1H),7.89(br,1H),7.77(s,1H),7.55(t,J＝8.0Hz,1H),7.36-7.31(m,3H),7.15(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.28(dd,J＝16.8Hz,1.6Hz,1H),6.04(br,1H),5.78(dd,J＝10.0Hz,1.6Hz,1H),3.78(s,3H),3.05(br,4H),2.44(t,J＝4.4Hz,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₂N₈O₃F,607.2581；found,607.2589.

N- (3- (6- (2-fluorophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 85)

Orange solid, yield 67%.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),8.85(s,1H),8.50(s,1H),7.86(s,1H),7.76(s,1H),7.64-7.68(t,J＝8.0Hz,1H),7.52-7.56(t,J＝8.0Hz,2H),7.31-7.35(t,J＝8.0Hz,3H),7.14(d,J＝8.0Hz,1H),6.54(s,1H),6.46(dd,J＝16.0Hz,8.0Hz,1H),6.27(d,J＝16.0Hz,1H),6.03(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.05(br,4H),2.43(s,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₂N₈O₃F,607.2503；found,607.2534.

N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6- (4-methoxyphenyl) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 88)

Orange solid, 65% yield, mp 296.5-297.3 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),8.84(s,1H),8.34(s,1H),8.26(d,J＝8.8Hz,1H),7.89(d,J＝7.2Hz,1H),7.74(s,1H),7.55(t,J＝8.0Hz,1H),7.36(d,J＝8.4Hz,1H),7.15(d,J＝8.4Hz,,1H),7.05(d,J＝9.2Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.06(br,1H),5.78(d,J＝10.0Hz,1.6Hz,1H),3.84(s,3H),3.78(s,3H),3.05(br,4H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₃₅N₈O₄,619.2781；found,619.2780.

N- (3- (6- (3, 5-difluorophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO. 89)

Red solid, yield 64%, mp 291.2-291.4 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),8.92(s,1H),8.57(s,1H),7.96(d,J＝7.2Hz,1H),7.87(br,1H),7.77(s,1H),7.54(t,J＝8.0Hz,1H),7.43-7.39(m,1H),7.33(s,1H),7.14(d,J＝7.2Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.00(br,1H),5.78(dd,J＝10.0Hz,1.6Hz,1H),3.77(s,3H),3.05(br,4H),2.43(br,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₁N₈O₃F₂,625.2487；found,625.2482.

N- (3- (6- (3, 4-difluorophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 90)

Orange solid, yield 67%, Mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.43(s,1H),8.89(s,1H),8.50(br,1H),8.28-8.23(m,1H),8.15(br,1H),7.87(s,1H),7.77(s,1H),7.61-7.52(m,2H),7.34(s,1H),7.14(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.0Hz,1H),6.27(dd,J＝16.8Hz,1.6Hz,1H),6.02(br,1H),5.78(dd,J＝10.0Hz,1.6Hz,1H),3.77(s,3H),3.05(br,4H),2.43(t,J＝4.4Hz,4H),2.22(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₃H₃₁N₈O₃F₂,625.2487；found,625.2478.

N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -6- (3-methoxyphenyl) -7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 91)

Brown solid, yield 52%, Mp >300 ℃.

¹H NMR(400MHz,DMSO-d₆):δ10.42(s,1H),8.88(s,1H),8.45(s,1H),7.89(s,1H),7.82-7.81(m,2H),7.76(s,1H),7.55(t,J＝8.0Hz,1H),7.41(t,J＝8.0Hz,1H),7.36-7.34(br,1H),7.16(d,J＝7.6Hz,1H),7.08(d,J＝8.4Hz,1H),6.54(s,1H),6.47(dd,J＝16.8Hz,10.4Hz,1H),6.28(d,J＝17.2Hz,1H),6.04(br,1H),5.80(m,1H),3.82(s,3H),3.78(s,3H),3.05(br,4H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₃₅N₈O₄,619.2781；found,619.2780.

N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-6-phenyl-7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 92)

Brown solid, yield 85%.

¹H NMR(400MHz,DMSO-d₆):δ10.40(s,1H),8.25-8.28(m,2H),8.20(s,1H),7.90(d,J＝8.0Hz,1H),7.72(s,1H),7.54(t,J＝8.0Hz,1H),7.48-7.50(m,3H),7.38(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.53(d,J＝4.0Hz,1H),6.46(dd,J＝16.0Hz,8.0Hz,1H),6.27(dd,J＝16.0Hz,4.0Hz,1H),6.03(br,1H),5.77(m,1H),3.82(s,3H),3.03(s,4H),2.72(s,3H),2.44(br,4H),2.23(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₃₅N₈O₄,603.2754；found,603.2750.

N- (3- (6- (4-fluorophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-8 (7H) pteridinyl) phenyl) acrylamide (SEQ ID NO: 93)

Brown solid, 73% yield.

¹H NMR(400MHz,DMSO-d₆):δ10.48(s,1H),8.33-8.37(m,2H),8.22(s,1H),7.91(d,J＝8.0Hz,1H),7.74(s,1H),7.54(t,J＝8.0Hz,1H),7.30-7.38(m,3H),7.12(d,J＝8.0Hz,1H),6.53(s,1H),6.48(dd,J＝16.0Hz,8.0Hz,1H),6.27(d,J＝16.0Hz,1H),6.01(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.06(s,4H),2.70(s,3H),2.53(s,4H),2.29(s,3H).HRMS(ESI)(m/z):[M+H]⁺calcd for C₃₄H₃₅N₈O₄,621.2738；found,603.2726.

The synthesis of the 1, 4-dihydro-2H-pyrimido [4,5-d ] [1,3] oxazine-2-one compounds of the invention is as follows:

reagents and conditions: (a) (3-aminophenyl) carbamic acid tert-butyl ester, DIPEA, CH₃CN, refluxing for 6 h; (b) LiAlH₄,THF,0℃,4h；(c)MnO₂,CH₂Cl₂At room temperature, overnight; (d) grignard reagent, THF,0 ℃,5 h; (e) CDI, K₂CO₃THF, reflux, overnight; (f) arylamine, trifluoroacetic acid and trifluoroethanol are refluxed for 24 hours; (g) trifluoroacetic acid, CH₂Cl₂Room temperature, 5 h; (h) acryloyl chloride, Et₃N,CH₂Cl₂From 0 ℃ to room temperature overnight.

Example 2

The specific synthesis method of the steps a-h is as follows:

synthesis of ethyl 4- ((3- ((tert-butoxycarbonyl) amino) phenyl) amino) -2-chloropyrimidine-5-carboxylate

Ethyl 2, 4-dichloro-5-pyrimidinecarboxylate (22.100g,100mmol) and DIPEA (12.900g,100mmol) were weighed into a 500mL single-neck flask, and dissolved in 100mL acetonitrile. Tert-butyl (3-aminophenyl) carbamate (20.800g,100mmol) was dissolved in 100mL of acetonitrile, added dropwise to the reaction mixture, and refluxed for 6 hours. TLC tracking till the raw material conversion, cooling to room temperature, suction filtering, washing with acetonitrile, and drying the filter cake to obtain 33.710g of 4- ((3- ((tert-butoxycarbonyl) amino) phenyl) amino) -2-chloropyrimidine-5-ethyl formate.

¹H NMR(400MHz,DMSO-d₆)δ10.23(s,1H),9.50(s,1H),8.80(s,1H),7.70(s,1H),7.35(d,J＝8.0Hz,1H),7.29(t,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),4.38(q,J＝7.2Hz,2H),1.49(s,9H),1.36(t,J＝7.2Hz,3H).LC-MS:m/z:393.1(M+H)⁺.

(3- ((2-chloro-5- (hydroxymethyl) pyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester

Ethyl 4- ((3- ((tert-butoxycarbonyl) amino) phenyl) amino) -2-chloropyrimidine-5-carboxylate (31.360g,80mmol) was weighed into a 5000mL two-necked flask, dissolved in 100mL anhydrous tetrahydrofuran, and stirred in an ice bath for 10 minutes. Lithium aluminum hydride (12.160g,320mmol) was dissolved in 150mL of anhydrous tetrahydrofuran, and the solution was added dropwise to the reaction mixture, followed by stirring in an ice bath for 4 hours. TLC tracks the conversion of the starting material and the reaction solution is added dropwise to 250mL of saturated NH in portions₄Extracting with ethyl acetate in Cl aqueous solution, collecting organic layer, and collecting anhydrous Na₂SO₄Drying and rotatingThe solvent was removed by evaporation. The crude product was isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate 2:1, v/v). To obtain 3.638g of tert-butyl (3- ((2-chloro-5- (hydroxymethyl) pyrimidin-4-yl) amino) phenyl) carbamate.

¹H NMR(400MHz,CDCl₃)δ8.38(s,1H),7.87(s,1H),7.70(s,1H),7.34(d,J＝8.4Hz,1H),7.25(t,J＝8.0Hz,1H),7.05(d,J＝8.0Hz,1H),6.66(s,1H),4.65(s,2H),1.52(s,9H).LC-MS:m/z:351.1(M+H)⁺.

(3- ((2-chloro-5-formylpyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester

Tert-butyl (3- ((2-chloro-5- (hydroxymethyl) pyrimidin-4-yl) amino) phenyl) carbamate (3.500g,10mmol) was weighed into a 100mL single-neck flask and dissolved by adding 40mL of dichloromethane. Manganese dioxide (58%, 15.000g,100mmol) was added portionwise and stirred at room temperature overnight. The conversion of the starting material was followed by TLC, filtered through celite, the filtrate was spin dried and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate 4:1, v/v). To obtain 2.850g of tert-butyl (3- ((2-chloro-5-formylpyrimidin-4-yl) amino) phenyl) carbamate.

¹H NMR(400MHz,CDCl₃)δ10.60(s,1H),9.89(s,1H),8.56(s,1H),7.82(t,J＝2.0Hz,1H),7.39(d,J＝8.8Hz,1H),7.31(t,J＝8.0Hz,1H),7.20(d,J＝8.0Hz,1H),6.58(s,1H),1.53(s,9H).LC-MS:m/z:349.1(M+H)⁺.

(3- ((2-chloro-5- (1-hydroxyethyl) pyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester

Tert-butyl (3- ((2-chloro-5-formylpyrimidin-4-yl) amino) phenyl) carbamate (1.044g,3mmol) was weighed into a 50mL two-necked flask, dissolved in 20mL anhydrous tetrahydrofuran,stirring in ice bath for 10 min under argon protection. Methyl magnesium bromide (1M in THF,9mL) was slowly added to the reaction solution, and stirred in an ice bath for 5 hours. TLC to follow the conversion of starting material, the reaction solution was poured into 30mL of saturated NH₄Extracting with ethyl acetate in Cl aqueous solution, collecting organic layer, and collecting anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate 2:1, v/v). To obtain 0.831g of tert-butyl (3- ((2-chloro-5- (1-hydroxyethyl) pyrimidin-4-yl) amino) phenyl) carbamate.

¹H NMR(400MHz,CDCl₃)δ8.83(s,1H),7.77(s,1H),7.70(s,1H),7.29(d,J＝8.4Hz,1H),7.23(t,J＝8.0Hz,1H),7.05(d,J＝8.0Hz,1H),6.67(s,1H),4.87(q,J＝6.4Hz,1H),1.55(d,J＝6.4Hz,3H),1.52(s,9H).LC-MS:m/z:365.1(M+H)⁺.

(3- (7-chloro-4-methyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester

Tert-butyl (3- ((2-chloro-5- (1-hydroxyethyl) pyrimidin-4-yl) amino) phenyl) carbamate (0.815g,2.2mmol), potassium carbonate (0.455g,3.3mmol), 1' -carbonyldiimidazole (1.069g,6.6mmol) were weighed into a 25mL single-neck flask, 10mL of anhydrous tetrahydrofuran was added, and refluxed overnight. TLC tracing raw material conversion, adding ice water, extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate 2:1, v/v). To obtain (3- (7-chloro-4-methyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester 0.754 g.

¹H NMR(400MHz,DMSO-d₆)δ9.59(s,1H),8.54(s,1H),7.61(s,1H),7.41-7.36(m,2H),6.98(d,J＝6.8Hz,1H),5.86(q,J＝6.4Hz,1H),1.74(d,J＝6.4Hz,3H),1.47(s,9H).LC-MS:m/z:391.1(M+H)⁺.

(3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-methyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester

Weighing (3- (7-chloro-4-methyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester (0.737g,1.89mmol), 2-methoxy-4- (4-methylpiperazin-1-yl) aniline (0.502g,2.27mmol) were dissolved in a 50mL two-necked flask by adding 15mL of trifluoroethanol, trifluoroacetic acid (210. mu.L, 2.84mmol) was added dropwise under argon protection, and the mixture was refluxed for 24 hours. TLC to follow the conversion of the starting material, cooling to room temperature and adding saturated NaHCO₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (dichloromethane/methanol ═ 30:1, v/v). To obtain (3- (7- ((2-methoxyl-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-methyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester 0.294 g%.

¹H NMR(400MHz,CDCl₃)δ8.46(s,1H),8.18(d,J＝8.8Hz,1H),7.79(s,1H),7.74(s,1H),7.24-7.20(m,3H),6.69(s,1H),6.54(d,J＝2.4Hz,1H),6.50(dd,J＝8.8Hz,J＝2.4Hz,1H),4.52(q,J＝6.8Hz,1H),3.85(s,3H),3.17(t,J＝4.4Hz,4H),2.60(t,J＝4.8Hz,4H),2.36(s,3H),1.51(s,9H),1.49(d,J＝6.0Hz,3H).LC-MS:m/z:576.3(M+H)⁺.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-methyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 94)

Weighing (3- (7- ((2-methoxy-4- (4-methylpiperazine))-1-yl) phenyl) amino) -4-methyl-2-oxo-2H-pyrimido [4,5-d][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester (0.277g,0.48mmol) in a 25mL single-neck flask was dissolved by addition of 6mL of dichloromethane, and 1mL of trifluoroacetic acid was added dropwise and stirred at room temperature for 5 hours. TLC followed conversion of starting material and saturated NaHCO was added₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying, rotary evaporating to remove solvent, and using crude product without separation and purification for next reaction.

The Boc removed product from the previous step (0.187g,0.39mmol) was dissolved in 5mL of dichloromethane, triethylamine (0.060g,0.6mmol) was added and stirred in ice bath for 10 min. Further, acryloyl chloride (42. mu.L, 0.51mmol) was dissolved in 1mL of methylene chloride, and the resulting solution was added to the reaction mixture, followed by stirring at room temperature overnight. TLC followed conversion of starting material and saturated NaHCO was added₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying, rotary evaporation to remove solvent, and separation and purification of the crude product by silica gel column chromatography (dichloromethane/methanol ═ 20:1, v/v). Obtaining N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-methyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) acrylamide 0.091 g.

¹H NMR(400MHz,DMSO-d₆)δ10.46(s,1H),8.23(s,1H),7.85(d,J＝8.0Hz,1H),7.81(s,1H),7.67(s,1H),7.47(t,J＝8.0Hz,1H),7.27(d,J＝8.4Hz,1H),7.10(d,J＝8.0Hz,1H),6.56(d,J＝1.2Hz,1H),6.48(dd,J＝16.8Hz,J＝9.6Hz,1H),6.27(dd,J＝17.2Hz,J＝1.6Hz,1H),6.11-6.09(m,1H),5.77(dd,J＝16.8Hz,J＝1.6Hz,1H),5.73(q,J＝6.4Hz,1H),3.76(s,3H),3.22-3.20(m,4H),3.02-2.99(m,4H),2.61(s,3H),1.70(d,J＝6.4Hz,3H).HRMS(ESI)(m/z):(M+H)⁺calcd for C₂₈H₃₂N₇O₄530.2516,found,530.2512.

The following compounds were synthesized according to the methods described above in steps a-g:

n- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-ethyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO. 95)

¹H NMR(400MHz,DMSO-d₆)δ10.46(s,1H),8.22(s,1H),7.85(d,J＝8.4Hz,1H),7.81(s,1H),7.66(s,1H),7.47(t,J＝8.0Hz,1H),7.26(d,J＝8.4Hz,1H),7.09(d,J＝8.0Hz,1H),6.55(d,J＝2.0Hz,1H),6.48(dd,J＝16.8Hz,J＝6.8Hz,1H),6.27(dd,J＝16.8Hz,J＝1.6Hz,1H),6.10-6.09(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.55(t,J＝6.8Hz,1H),3.76(s,3H),3.19(t,J＝4.4Hz,4H),2.92(t,J＝4.4Hz,4H),2.56(s,3H),2.09-1.95(m,2H),1.03(t,J＝7.2Hz,3H).HRMS(ESI)(m/z):(M+H)⁺calcd for C₂₉H₃₄N₇O₄544.2672,found,544.2654.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-propyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 96)

¹H NMR(400MHz,DMSO-d₆)δ10.45(s,1H),8.22(s,1H),7.85(d,J＝8.0Hz,1H),7.80(s,1H),7.66(s,1H),7.47(t,J＝8.0Hz,1H),7.25(d,J＝8.4Hz,1H),7.08(d,J＝8.0Hz,1H),6.54(d,J＝2.0Hz,1H),6.48(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),6.10-6.09(m,1H),5.77(dd,J＝9.6Hz,J＝1.6Hz,1H),5.60(t,J＝7.2Hz,1H),3.76(s,3H),3.14(t,J＝4.4Hz,4H),2.79(t,J＝4.4Hz,4H),2.47(s,3H),2.00-1.92(m,2H),1.56-1.44(m,2H),0.99(t,J＝7.2Hz,3H).HRMS(ESI)(m/z):(M+H)⁺calcd forC₃₀H₃₆N₇O₄558.2829,found,558.2836.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4-isopropyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 97)

¹H NMR(400MHz,DMSO-d₆)δ10.52(s,1H),8.22(s,1H),7.87(d,J＝8.0Hz,1H),7.85(s,1H),7.64(s,1H),7.47(t,J＝8.0Hz,1H),7.26(d,J＝7.6Hz,1H),7.05(d,J＝7.6Hz,1H),6.57(d,J＝1.6Hz,1H),6.50(dd,J＝16.8Hz,J＝10.0Hz,1H),6.26(dd,J＝16.8Hz,J＝1.6Hz,1H),6.14-6.10(m,1H),5.77(dd,J＝10.0Hz,J＝1.6Hz,1H),5.40(d,J＝4.8Hz,1H),3.77(s,3H),3.18(t,J＝4.4Hz,4H),2.74(s,3H),2.27-2.22(m,1H),1.04(d,J＝6.8Hz,3H),0.99(d,J＝6.8Hz,3H).HRMS(ESI)(m/z):(M+H)⁺calcd for C₃₀H₃₆N₇O₄558.2829,found,558.2831.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 98)

¹H NMR(400MHz,CDCl₃+CD₃OD)δ8.09(s,1H),7.89(s,1H),7.75(d,J＝8.4Hz,1H),7.47(t,J＝8.0Hz,1H),7.42-7.39(m,1H),7.07(d,J＝7.6Hz,1H),6.42-6.38(m,2H),6.15(d,J＝7.2Hz,1H),5.71(dd,J＝9.2Hz,J＝2.8Hz,1H),5.36(s,2H),3.80(s,3H),3.37(t,J＝4.8Hz,4H),3.24(t,J＝4.8Hz,4H),2.82(s,3H).HRMS(ESI)(m/z):(M+H)⁺calcdforC₂₇H₃₀N₇O₄516.2359,found,516.2364.

The specific synthesis method of the compound 006-008 comprises the following steps:

reagent and stripA piece: (a) grignard reagent, THF,0 ℃,6 h; (b) CDI, K₂CO₃THF, reflux, overnight; (c) arylamine, trifluoroacetic acid and trifluoroethanol are refluxed for 24 hours; (d) trifluoroacetic acid, CH₂Cl₂Room temperature, 5 h; (e) acryloyl chloride, Et₃N,CH₂Cl₂From 0 ℃ to room temperature overnight.

(3- ((2-chloro-5- (2-hydroxyprop-2-yl) pyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester

Ethyl 4- ((3- ((tert-butoxycarbonyl) amino) phenyl) amino) -2-chloropyrimidine-5-carboxylate (2.352g,6mmol) was weighed into a 50mL two-necked flask, dissolved in 20mL anhydrous tetrahydrofuran under argon and stirred in an ice bath for 10 minutes. Methyl magnesium bromide (1M in THF,24mL) was slowly added to the reaction solution, and stirred for 6 hours in an ice bath. TLC to follow the conversion of starting material, the reaction solution was poured into 50mL of saturated NH₄Extracting with ethyl acetate in Cl aqueous solution, collecting organic layer, and collecting anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate 2.5:1, v/v). To obtain 1.586g of tert-butyl (3- ((2-chloro-5- (2-hydroxypropan-2-yl) pyrimidin-4-yl) amino) phenyl) carbamate.

¹H NMR(400MHz,DMSO-d₆)δ10.01(s,1H),9.41(s,1H),8.12(s,1H),7.62(t,J＝2.0Hz,1H),7.40(dd,J＝8.0Hz,J＝1.2Hz,1H),7.25(t,J＝8.4Hz,1H),7.13(d,J＝8.8Hz,1H),6.43(s,1H),1.56(s,6H),1.48(s,9H).LC-MS:m/z:379.1(M+H)⁺.

(3- (7-chloro-4, 4-dimethyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester

Weighing (3- ((2-chloro-5-) (2-hydroxypropan-2-yl) pyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester (1.512g,4mmol), potassium carbonate (0.828g,6mmol), 1' -carbonyldiimidazole (1.296g,8mmol) in a 25mL single neck flask, 10mL of anhydrous tetrahydrofuran was added and refluxed overnight. TLC tracing raw material conversion, adding ice water, extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (petroleum ether/ethyl acetate 2.5:1, v/v). To obtain (3- (7-chloro-4, 4-dimethyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester 1.049 g.

¹H NMR(400MHz,DMSO-d₆)δ9.57(s,1H),8.64(s,1H),7.57(s,1H),7.41-7.36(m,2H),7.01-6.99(m,1H),1.79(s,6H),1.47(s,9H).LC-MS:m/z:405.1(M+H)⁺.

(3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-dimethyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester

Weighing (3- (7-chloro-4, 4-dimethyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester (1.010g,2.5mmol), 2-methoxy-4- (4-methylpiperazin-1-yl) aniline (0.663g,3mmol) were dissolved in a 50mL two-necked flask by adding 15mL of trifluoroethanol, trifluoroacetic acid (280. mu.L, 3.77mmol) was added dropwise under argon protection, and the mixture was refluxed for 24 hours. TLC to follow the conversion of the starting material, cooling to room temperature and adding saturated NaHCO₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying and rotary evaporation to remove the solvent. The crude product was isolated and purified by silica gel column chromatography (dichloromethane/methanol 25:1, v/v). To obtain (3- (7- ((2-methoxyl-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-dimethyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester 0.516 g.

¹H NMR(400MHz,CDCl₃)δ8.07(s,1H),7.51(d,J＝8.4Hz,2H),7.47-7.43(m,2H),7.01(d,J＝7.2Hz,1H),6.45(s,1H),6.44(d,J＝2.4Hz,1H),6.18-6.16(m,1H),3.82(s,3H),3.16(t,J＝4.4Hz,4H),2.67(t,J＝4.4Hz,4H),2.42(s,3H),1.80(s,6H),1.49(s,9H).LC-MS:m/z:590.4(M+H)⁺.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-dimethyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO. 99)

Weighing (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-dimethyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) carbamic acid tert-butyl ester (0.500g,0.85mmol) was dissolved in a 25mL single-neck flask by adding 6mL of dichloromethane, and 1mL of trifluoroacetic acid was added dropwise thereto and stirred at room temperature for 5 hours. TLC followed conversion of starting material and saturated NaHCO was added₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying, rotary evaporating to remove solvent, and using crude product without separation and purification for next reaction.

The Boc-removed product from the previous step (0.335g,0.68mmol) was dissolved in 5mL of dichloromethane, triethylamine (0.102g,1.02mmol) was added, and the mixture was stirred in ice bath for 10 min. Further, acryloyl chloride (72. mu.L, 0.88mmol) was dissolved in 1mL of methylene chloride, and the resulting solution was added to the reaction mixture, followed by stirring at room temperature overnight. TLC followed conversion of starting material and saturated NaHCO was added₃The aqueous solution is neutralized to be alkaline. Extracting with dichloromethane, collecting organic layer, anhydrous Na₂SO₄Drying, rotary evaporation to remove solvent, and separation and purification of the crude product by silica gel column chromatography (dichloromethane/methanol ═ 20:1, v/v). Obtaining N- (3- (7- ((2-methoxyl-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-dimethyl-2-oxo-2H-pyrimido [4, 5-d)][1,3]Oxazin-1 (4H) -yl) phenyl) acrylamide 0.165 g.

¹H NMR(400MHz,CDCl₃)δ8.08(s,1H),7.79(d,J＝8.0Hz,1H),7.74(s,1H),7.47(t,J＝8.0Hz,1H),7.38-7.36(m,1H),7.05(d,J＝7.6Hz,1H),6.41(d,J＝2.0Hz,1H),6.36-6.33(m,2H),6.13(s,1H),5.70(dd,J＝9.2Hz,J＝2.4Hz,1H),3.80(s,3H),3.17(t,J＝4.4Hz,4H),2.80(t,J＝4.4Hz,4H),2.50(s,3H),1.80(s,6H).HRMS(ESI)(m/z):(M+H)⁺calcd forC₂₉H₃₄N₇O₄544.2672,found,544.2698.

The following 007 and 008 compounds were synthesized according to the methods described above for steps a-e:

n- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-diethyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 100)

¹H NMR(400MHz,CDCl₃)δ9.42(s,1H),7.96(s,1H),7.89(s,1H),7.81(d,J＝5.6Hz,1H),7.53(s,1H),7.42(t,J＝8.0Hz,1H),7.33-7.31(m,1H),6.99(d,J＝7.6Hz,1H),6.55-6.49(m,1H),6.36-6.32(m,2H),6.12(d,J＝7.6Hz,1H),5.65(d,J＝10.4Hz,1H),3.78(s,3H),3.41(t,J＝4.4Hz,4H),3.21(t,J＝4.4Hz,4H),2.80(s,3H),2.11-1.96(m,4H),1.00(t,J＝7.2Hz,6H).HRMS(ESI)(m/z):(M+H)⁺calcd forC₃₁H₃₈N₇O₄572.2985,found,572.2981.

N- (3- (7- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -4, 4-diisopropyl-2-oxo-2H-pyrimido [4,5-d ] [1,3] oxazin-1 (4H) -yl) phenyl) acrylamide (SEQ ID NO: 101)

¹H NMR(400MHz,CDCl₃)δ8.06(s,1H),7.96(s,1H),7.77(d,J＝4.8Hz,1H),7.49(s,1H),7.41(t,J＝8.0Hz,1H),6.97(d,J＝8.0Hz,1H),6.40(d,J＝2.0Hz,1H),6.36(dd,J＝16.8Hz,J＝1.2Hz,1H),6.18-6.12(m,2H),5.68(dd,J＝10.4Hz,J＝1.2Hz,1H),3.79(s,3H),3.09(t,J＝4.4Hz,4H),2.58(t,J＝4.8Hz,4H),2.36(s,3H),2.07-2.00(m,2H),1.96-1.89(m,2H),1.54-1.38(m,4H),0.97(t,J＝7.2Hz,6H).HRMS(ESI)(m/z):(M+H)⁺calcdforC₃₃H₄₂N₇O₄600.3298,found,600.3297.

The specific synthesis method of the compound 132-133 is as follows:

reagents and conditions: (a) r¹NH₂DIPEA,1, 4-dioxane, r.t.; (b) r²H₂DIPEA,1, 4-dioxane, r.t.; (c) Pd/C, H2, EtOH; (d) YCOCOOEt, HOAc, EtOH, reflux.

In the above preparation schemes, R, R1, R2 and Y refer to the definitions of the corresponding groups above. The compounds of the present invention can be prepared by those skilled in the art according to the actual preparation needs, using various starting compounds conventionally obtained in the art as starting materials.

Example 3

The specific synthesis method of the steps a-d is as follows:

synthesis of tert-butyl (4- (2-chloro-6-methyl-5-nitropyrimidin-4-amino) phenyl) carbamate

Weighing 2.07g (10mmol) of 2, 4-dichloro-6-methyl-5-nitropyrimidine and 2.07g (15mmol) of potassium carbonate, placing the materials in a 250mL round-bottom flask, adding 100mL of dichloromethane, mechanically stirring at normal temperature, weighing 2.08g (10mmol) of tert-butyl (3-aminophenyl) carbamate, dissolving in 50mL of dichloromethane, slowly dropwise adding into the reaction, continuing to stir for about 1 hour under an ice bath condition after the dropwise adding is finished, tracking by TLC until the raw materials are completely converted, directly pumping, reducing pressure, rotationally evaporating to remove the solvent, recrystallizing by using ethanol, pumping a filter cake, adding 1000mL of water for pulping, pumping and washing by using 500mL of water again to obtain 3.41g of orange solid of the relatively pure compound (4- (2-chloro-6-methyl-5-nitropyrimidine-4-amino) phenyl) tert-butyl carbamate, the yield was about 90%.

¹H NMR(400MHz,DMSO-d₆):δ9.91(s,1H),9.43(s,1H),7.64(s,1H),7.27-7.31(m,1H),7.23-7.24(m,2H),2.55(s,3H),1.48(s,9H)。

Synthesis of tert-butyl (3- ((2- ((2-methoxy-4- (4-methylpiperazino) phenyl) amino) -6-methyl-5-nitropyrimidin-4-yl) amino) phenyl) carbamate

2.21g (10mmol) of 2-methoxy-4- (4-methyl-1-piperazine) aniline, 2.58g (20mmol) of N, N-diisopropylethylamine, 3.79g (10mmol) of tert-butyl (3- (2-chloro-5-nitropyrimidin-4-amino) phenyl) carbamate were weighed into a 250ml round-bottomed flask, dissolved in 100ml of tetrahydrofuran, under argon protection, heated to reflux overnight, and followed by TLC until complete conversion of the starting material. Removing about 70ml of solvent by rotary evaporation, filtering out a solid by suction, washing a filter cake by 50ml of tetrahydrofuran, removing most of the solvent by rotary evaporation of a filtrate, pouring into water, separating out a reddish brown solid, filtering by suction, washing the filter cake by deionized water, drying, and purifying by dichloromethane/methanol to obtain 91g of the reddish brown solid of tert-butyl (3- ((2- ((2-methoxy-4- (4-methylpiperazinyl) phenyl) amino) -6-methyl-5-nitropyrimidin-4-yl) amino) phenyl) carbamate, wherein the yield is about 71%.

¹H NMR(400MHz,DMSO-d₆):δ10.27(s,1H),9.42(s,1H),8.86(s,1H),7.52(s,1H),7.39(d,J＝8.0Hz,1H),7.19(s,2H),7.09(s,1H),6.60(s,1H),6.32(s,1H),3.76(s,3H),3.42(s,3H),3.15(s,4H),2.61(s,4H),2.27(s,3H),1.47(s,9H)。

Synthesis of tert-butyl (3- ((5-amino-2- ((2-methoxy-4- (4-methylpiperazino) phenyl) amino) -6-methylpyrimidin-4-yl) amino) phenyl) carbamate

Tert-butyl (3- ((2- ((2-methoxy-4- (4-methylpiperazino) phenyl) amino) -6-methyl-5-nitropyrimidin-4-yl) amino) phenyl) carbamate (5.64g,10mmol) and palladium on carbon catalyst (0.56g, 0.5mmol, 10% Pd) were weighed into a 500mL round-bottomed flask, dissolved by adding 100mL of methanol and 200mL of dichloromethane, and reacted at room temperature for 12 hours with introduction of hydrogen gas. TLC tracks the conversion of the raw material, suction filtration is carried out, the filtrate is dried in a rotary mode, and the crude product is recrystallized by ethanol to obtain (3- ((5-amino-2- ((2-methoxy-4- (4-methylpiperazino) phenyl) amino) -6-methylpyrimidin-4-yl) amino) phenyl) carbamic acid tert-butyl ester white solid 4.86g, the yield is 91%.

¹H NMR(400MHz,DMSO-d₆)δ10.27(s,1H),9.42(s,1H),8.87(d,J＝8.0Hz,1H),7.53(s,1H),7.39(d,J＝8.0Hz,1H),7.32(d,J＝8.0Hz,1H),7.16(t,J＝8.0Hz,1H),7.06(d,J＝8.0Hz,1H),6.60(d,J＝2.4Hz,1H),6.38(dd,J＝8.8Hz,J＝2.4Hz,1H),4.39(s,2H),3.76(s,3H),3.42(s,3H),3.07(t,J＝4.4Hz,4H),2.48(t,J＝4.4Hz,4H),2.25(s,3H),1.48(s,9H).

Synthesis of tert-butyl (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) carbamate

Weighing 2.67g (5mmol) of tert-butyl (3- ((5-amino-2- ((2-methoxy-4- (4-methylpiperazino) phenyl) amino) -6-methylpyrimidin-4-yl) amino) phenyl) carbamate in a 250mL round-bottom flask, adding 10mL of glacial acetic acid and 150mL of anhydrous ethanol, then adding 890mg (5mmol) of ethyl benzoylformate, heating to reflux and stirring for about 8h, and TLC tracking until the starting material is completely converted. After the reaction is finished, the solvent is removed by rotary evaporation, a small amount of ethanol is added for dissolution, the filtration is carried out, and a filter cake is washed by ethanol, ammonia water and deionized water and dried. If impurities are present, the mixture is washed by heating with ethanol to obtain tert-butyl (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) carbamate as an orange-red solid (2.2 g, yield 70%).

¹H NMR(400MHz,DMSO-d₆):δ9.64(s,1H),8.24-8.26(m,2H),8.21(s,1H),7.59(s,1H),7.56(d,J＝8.0Hz,1H),7.47-7.50(m,3H),7.44(d,J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.00(d,J＝8.0Hz,1H),6.54(s,1H),6.07(br,1H),3.79(s,3H),3.07(br,4H),2.71(s,3H),2.46(br,4H),2.24(s,3H),1.45(s,9H).

Synthesis of 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-6-phenyl-7 (8h) -pteridinone

6.48g (10mmol) of tert-butyl (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) carbamate are weighed into a 250mL round-bottom flask, 100mL of dichloromethane are added, stirring is carried out at 0 ℃ and 25mL of trifluoroacetic acid are added dropwise. Stirring was then continued for 0.5h in ice bath, 2h at room temperature and TLC tracking was done until complete conversion of starting material. After the reaction is finished, the solvent is directly removed by rotary evaporation, water is added for dissolution, a saturated sodium bicarbonate solution is neutralized until the solution is alkaline, a large amount of solid is separated out, the filtration is carried out, a filter cake is washed by deionized water and dried, and 4.99g of 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-6-phenyl-7 (8h) -pteridinone red solid is obtained, wherein the yield is 91%.

1H NMR(400MHz,DMSO-d₆):δ9.64(s,1H),8.24-8.26(m,2H),8.21(s,1H),7.59(s,1H),7.56(d,J＝8.0Hz,1H),7.47-7.50(m,3H),7.44(d,J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.00(d,J＝8.0Hz,1H),6.54(s,1H),5.32(s,2H),3.79(s,3H),3.07(br,4H),2.71(s,3H),2.46(br,4H),2.24(s,3H),1.45(s,9H).

Synthesis of N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 102)

Weighing 1.4g (2.55mmol) of 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-6-phenyl-7 (8h) -pteridinone, placing the mixture in a 100mL round-bottom flask, adding 3mL of N-methylpyrrolidone, stirring in ice bath, dissolving 275mg (3.06mmol) of acryloyl chloride in 20mL of acetonitrile, dropwise adding the mixture into the reaction solution, stirring in ice bath for 0.5h after the dropwise adding is completed, stirring at room temperature for 3h, and tracking by TLC until the raw materials are completely converted. And (3) performing rotary evaporation on the reaction liquid to remove the solvent, then dropwise adding the reaction liquid into a sodium bicarbonate aqueous solution, separating out a red solid, performing suction filtration, washing a filter cake with deionized water, drying, and purifying dichloromethane/methanol to obtain 1.0g of N- (3- (2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-6-phenyl-8 (7H) -pteridinyl) phenyl) acrylamide orange red solid with the yield of 67%.

1H NMR(400MHz,DMSO-d₆):δ10.40(s,1H),8.25-8.27(m,2H),8.20(s,1H),7.90(d,J＝8.0Hz,H),7.72(s,1H),7.54(t,J＝8.0Hz,1H),7.48-7.50(m,3H),7.38(d,J＝8.0Hz,1H),7.12(d,J＝8.0Hz,1H),6.53(d,J＝4.0Hz,1H),6.46(q,J＝8.0,1H),6.27(dd,J＝8.0,4.0Hz,1H),6.03(br,1H),5.78(dd,J＝8.0,4.0Hz,1H),3.78(s,3H),3.03(br,4H),2.72(s,3H),2.44(t,J＝4.0Hz,4H),2.23(s,3H).

Synthesis of N- (3- (6- (4-fluoro-phenyl) -2- ((2-methoxy-4- (4-methyl-1-piperazinyl) phenyl) amino) -4-methyl-7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (SEQ ID NO: 103)

Reddish brown solid, yield 61%.

1H NMR(400MHz,DMSO-d₆):δ10.48(s,1H),8.33-8.37(m,2H),8.22(s,1H),7.91(d,J＝8.0Hz,H),7.74(s,1H),7.54(t,J＝8.0Hz,1H),7.30-7.38(m,3H),7.12(d,J＝8.0Hz,1H),6.53(s,1H),6.48(q,J＝8.0,1H),6.27(dd,J＝8.0,2.0Hz,1H),6.01(br,1H),5.78(d,J＝8.0Hz,1H),3.77(s,3H),3.06(br,4H),2.70(s,3H),2.53(br,4H),2.29(s,3H).

The specific synthesis method of the compound 134-137 is as follows:

reagents and conditions: (a) r¹NH₂DIPEA, acetonitrile, reflux, 6 h; (b) lithium aluminum hydride, THF,0 ℃,4 h.; (c) manganese dioxide, dichloromethane and 6 hours; (d) grignard reagent, THF,0 ℃,4 h; (e) manganese dioxide, manganese dioxide and dichloromethane for 6 hours; (f) YCH2COOEt, K₂CO₃DMF,8 h; (g) m-CPBA, dichloromethane and 12 h; (h) r²NH₂,TFA,2-Butanol,110℃。

In the above preparation process, R¹、R²、R⁵And Y refer to the corresponding group definitions above. The compounds of the present invention can be prepared by those skilled in the art according to the actual preparation needs, using various starting compounds conventionally obtained in the art as starting materials.

Example 4

The specific synthesis method of the steps a-d is as follows:

synthesis of ethyl 4- (3-tert-butoxycarbonylamino aniline) -2-methylthiopyrimidine-5-carbonate

Weighing 2.33g (10mmol) of 4-chloro-2-methylthio pyrimidine-5-ethyl carbonate, 2.08g (10mmol) of tert-butyl (3-aminophenyl) carbamate, 2.58g (20mmol) of DIPEA and placing in a 250ml flask, adding 80ml of acetonitrile for dissolving, then heating and refluxing, reacting for about 4 hours, tracking by TLC until the raw materials are completely converted, stopping the reaction, cooling to room temperature, directly filtering, washing a filter cake with cold acetonitrile for three times, and drying the filter cake in an infrared oven to obtain 3.84g of pure ethyl 4- (3-tert-butoxycarbonylamino aniline) -2-methylthio pyrimidine-5-carbonate as a white solid, wherein the yield is 95%.

¹H NMR(400MHz,CDCl₃)δ10.37(s,1H),8.76(s,1H),7.90(s,1H),7.34(d,J＝8.0Hz,1H),7.24(t,J＝8.0Hz,1H),7.02(d,J＝8.0Hz,1H),6.53(s,1H),4.38(q,J＝8.0Hz,J＝16.0Hz,2H),2.55(s,3H),1.52(s,9H),1.40(t,J＝8.0Hz,3H).

Synthesis of (tert-butyl-3- (5- (hydroxymethyl) -2- (methylthio) pyrimidine-4-substituted amino) benzamide carbonate

Weighing 2.02g (5mmol) of 4- (3-tert-butyloxycarbonylaminoaniline) -2-methylthiopyrimidine-5-carbonic acid ethyl ester, placing the mixture in a 250ml flask, adding 50ml of anhydrous tetrahydrofuran for dissolving, and slowly dropping 20ml of LiAlH under the ice bath condition₄(1M inTHF), after the dropwise addition, the reaction is continued for 0.5 hour under the ice bath condition, TLC tracks until the raw materials are completely converted, saturated ammonium chloride is added into the reaction to stop the reaction, tetrahydrofuran is removed by rotary evaporation, then ethyl acetate is used for extraction, saturated brine is used for washing, anhydrous sodium sulfate is dried, solvent is removed by rotary evaporation, and the crude product is separated by silica gel column chromatography (petroleum ether/ethyl acetate is 3:1, v/v) to obtain (tert-butyl-3- (5- (hydroxymethyl) -2- (methylthio) pyrimidine-4-substituted amino) benzamide carbonate, namely 760mg of white solid, and the yield is 42%.

¹H NMR(400MHz,CDCl₃)δ8.02(s,1H),7.86(s,1H),7.80(s,1H),7.36(dd,J＝4.0,8.0Hz,1H),7.22(t,J＝8.0Hz,1H),6.95(dd,J＝4.0,8.0Hz,1H),6.52(s,1H),4.61(s,2H),2.52(s,3H),1.52(s,9H).

Synthesis of tert-butyl 3- ((5-formyl-2- (methylthio) pyrimidin-4-substituted) amino) phenyl) carbamate

362mg (1mmol) of (tert-butyl-3- (5- (hydroxymethyl) -2- (methylthio) pyrimidine-4-substituted amino) benzamide carbonic ester is weighed into a 100ml flask, 50ml of dichloromethane is added for dissolution, then 870mg (10mmol) of active manganese dioxide is added and stirred for about 4 hours at normal temperature, TLC is used for tracking till the raw materials are completely converted, diatomite is filtered in a suction manner, the filtrate is rotated and evaporated to remove the solvent, and the crude product is separated by silica gel column chromatography (petroleum ether/ethyl acetate is 10:1, v/v) to obtain 306mg of 3- ((5-formyl-2- (methylthio) pyrimidine-4-substituted) amino) phenyl) carbamic acid tert-butyl ester pale yellow solid with the yield of 85 percent.

¹H NMR(400MHz,CDCl₃)δ10.61(s,1H),9.77(s,1H),8.43(s,1H),7.98(s,1H),7.36(dd,J＝4.0,8.0Hz,1H),7.25-7.29(m,1H),7.03(dd,J＝4.0,8.0Hz,1H),6.51(s,1H),2.59(s,3H),1.53(s,9H).

Synthesis of tert-butyl 3- (2- (methylthio) -7-oxopyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) carbamate

360mg (1mmol) of tert-butyl 3- ((5-formyl-2- (methylthio) pyrimidin-4-substituted) amino) phenyl) carbamate and (EtO)₂P(O)CH₂CO₂Et in 100ml flask, adding 50ml anhydrous THF to dissolve, then adding 48mg NaH (2mmol) under ice bath condition, stirring at room temperature for about 4 hours after adding, TLC tracking till the raw material is completely converted, slowly dropping water to quench, rotary evaporating filtrate to remove solvent, purifying the crude product with silica gel columnChromatography (petrol ether/ethyl acetate 20:1, v/v) separation to give 3- (2- (methylthio) -7-oxopyrido [2, 3-d)]Pyrimidine-8 (7H) -substituted) phenyl) carbamic acid tert-butyl ester 270mg as a yellow solid in 71% yield.

¹H NMR(500MHz,CDCl₃)δ9.10(s,1H),8.09(dt,J＝7.5,2.0Hz,1H),7.72(t,J＝2.0Hz,1H),7.56(d,J＝11.0Hz,1H),7.41(t,J＝7.4Hz,1H),7.34(dt,J＝7.5,2.0Hz,1H),6.65(s,1H),6.44(d,J＝10.8Hz,1H),2.54(s,3H),1.50(s,9H).

Synthesis of tert-butyl 3- (2- (methylsulfonyl) -7-oxopyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) carbamate

Reacting 3- (2- (methylsulfonyl) -7-oxo-pyrido [2,3-d ]]Pyrimidine-8 (7H) -substituted) phenyl) carbamic acid tert-butyl ester 922mg (2.4mmol) is placed in a 100ml flask, 60ml dichloromethane is added for dissolution, 1.24g (7.2mmol) of m-CPBA is added in portions under ice bath condition within 30min, after the addition, the temperature is slowly raised to room temperature for stirring for about 8 hours, TLC is followed until the raw material is completely converted, 30ml saturated Na is added₂S₂O₃Stirring for half an hour, extracting with dichloromethane, drying with anhydrous sodium sulfate, rotary evaporating to remove solvent, and separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate 10:1, v/v) to obtain 3- (2- (methylsulfonyl) -7-oxopyrido [2,3-d ]]Pyrimidine-8 (7H) -substituted) phenyl) carbamic acid tert-butyl ester as a white solid 808mg, 81% yield.

¹H NMR(500MHz,CDCl3)δ9.17(s,1H),8.30(t,J＝2.0Hz,1H),7.61-7.55(m,2H),7.39(t,J＝7.5Hz,1H),7.16(dt,J＝7.5,2.0Hz,1H),6.63(s,1H),6.44(d,J＝10.8Hz,1H),3.25(s,3H),1.50(s,9H).

Synthesis of tert-butyl (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxopyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) carbamate

808mg (1.94mmol) of tert-butyl 3- (2- (methylthio) -7-oxopyrido [2,3-d ] pyrimidine-8 (7h) -substituted) phenyl) carbamate and 430mg (1.94mmol) of 2-methoxy-4- (4-methylpiperazine-1-substituted) aniline in a 100ml flask were dissolved in 50ml of sec-butanol, 145. mu.l (1.94mmol) of TFA were added, the mixture was stirred at 110 ℃ for about 12 hours, TLC followed until the starting material had completely converted, the solvent was removed by rotary evaporation, the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 30:1, v/v) to give (3- (2- ((2-methoxy-4- (4-methylpiperazine-1-yl) phenyl) amino) -7-oxopyrido [2,3-d ] pyrimidin-8 (7H) -substituted) phenyl) carbamic acid tert-butyl ester 756mg as a yellow solid in 70% yield.

¹H NMR(500MHz,CDCl₃)δ8.71(s,1H),8.53(t,J＝1.9Hz,1H),7.56(d,J＝10.8Hz,1H),7.46-7.35(m,2H),7.24(dt,J＝7.3,2.1Hz,1H),6.91(d,J＝7.5Hz,1H),6.60(s,1H),6.44(d,J＝10.8Hz,1H),6.40–6.31(m,2H),5.10(s,1H),3.94(s,3H),3.20(t,J＝5.2Hz,4H),2.98(t,J＝5.1Hz,4H),2.60(s,3H),1.50(s,9H).

Synthesis of 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) pyrido [2,3-d ] pyrimidin-7 (8h) -one

756mg (1.36mmol) of tert-butyl (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxopyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) carbamate was dissolved in 16ml of dichloromethane in a 50ml flask, 4ml of TFA was added and stirred at room temperature for 4h, TLC was followed until the starting material was completely converted, the solvent was removed by rotary evaporation, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 30:1, v/v) to give 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) pyrido [2,3-d ] pyrimidin-7 (8-hydro) -one 571mg as a yellow solid, yield 92%.

¹H NMR(400MHz,DMSO-d₆)δ8.71(s,1H),8.08(s,1H),7.87(d,J＝9.6,1H),7.45(d,J＝8.8,1H),7.18(t,J＝8.0Hz,1H),6.71(d,J＝8.4Hz,1H),6.55(d,J＝2.4Hz,2H),6.36-6.41(m,3H),6.15(br,1H),5.25(br,2H),3.79(s,3H),3.06(t,J＝4.8Hz,4H),2.45(t,J＝4.8Hz,4H),2.23(s,3H).

Synthesis of N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxopyrido [2,3-d ] pyrimidin-8 (7H) -substituted) phenyl) acrylamide (SEQ ID NO: 104)

571mg (1.25mmol) of 8- (3-aminophenyl) -2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) pyrido [2,3-d ] pyrimidin-7 (8h) -one is put in a 50ml flask, 20ml of anhydrous dichloromethane is added to dissolve the solvent, 225mg (2.5mmol) of acryloyl chloride is slowly added dropwise into the system at 0 ℃, stirring is continued for 4h after the dropwise addition, TLC tracks till the raw material is completely converted, the solvent is removed by rotary evaporation, the crude product is separated by silica gel column chromatography (dichloromethane/methanol ═ 30:1, v/v) to obtain N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxopyrido [2,3-d ] pyrimidin-8 (7H) -substituted) phenyl) acrylamide 316mg as a yellow solid in 62% yield.

¹H NMR(400MHz,DMSO-d₆)δ10.34(s,1H),8.74(s,1H),8.15(s,1H),7.86-7.92(m,2H),7.60(s,1H),7.51(t,J＝8.0Hz,1H),7.29(d,J＝8.8Hz,1H),7.01(d,J＝8.0Hz,1H),6.52(d,J＝1.6Hz,1H),6.40-6.47(m,2H),6.28(dd,J＝1.6,17.2Hz,1H),6.02(br,1H),5.78(m,1H),3.77(s,3H),3.03(m,4H),2.44(m,4H),2.03(s,3H).

Synthesis of N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -5-methyl-7-oxopyrido [2,3-d ] pyrimidin-8 (7H) -substituted) phenyl) acrylamide (SEQ ID NO: 105)

¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.80(s,1H),8.09(s,1H),7.89(d,J＝6.8Hz,1H),7.56(s,1H),7.50(t,J＝8.0Hz,1H),7.28(d,J＝8.8Hz,1H),6.98(d,J＝7.6Hz,1H),6.51(s,1H),6.47(dd,J＝1.6,17.2Hz,1H),6.32(s,1H),6.27(d,J＝6.8Hz,1H),6.01(br,1H),5.78(m,1H),3.78(s,3H),3.03(m,4H),2.43-2.46(m,7H),2.03(s,3H).

Synthesis of N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -7-oxo-5-phenylpyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) acrylamide (SEQ ID NO: 106)

¹H NMR(400MHz,DMSO-d₆)δ10.45(s,1H),8.42(s,1H),8.27(s,1H),7.90(s,1H),7.52-7.67(m,8H),7.29(d,J＝8.8Hz,1H),7.07(d,J＝7.6Hz,1H),6.57(s,1H),6.51(dd,J＝10.0,16.8Hz,1H),6.39(s,1H),6.29(dd,J＝2.0,10.0Hz,1H),6.10(br,1H),5.79(dd,J＝2.0,16.8Hz 1H),3.78(s,3H),2.96(m,4H),2.43(s,3H).

Synthesis of N- (3- (2- ((2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) amino) -5-methyl-7-oxo-6-phenylpyrido [2,3-d ] pyrimidin-8 (7h) -substituted) phenyl) acrylamide (SEQ ID NO: 107)

¹H NMR(400MHz,DMSO-d₆)δ10.34(s,1H),8.89(s,1H),8.11(s,1H),7.87(d,J＝8.0Hz,1H),7.64(s,1H),7.51(t,J＝8.0Hz,1H),7.44(t,J＝7.2Hz,1H),7.34-7.38(m,1H),7.29-7.31(m,3H),7.04(d,J＝8.0Hz,1H),6.53(d,J＝1.6Hz,1H),6.47(dd,dd,J＝17.2,10.0Hz,1H),6.27(dd,J＝1.6,16.8Hz,1H),6.03(br,1H),5.78(dd,J＝1.6,10.0Hz 1H),3.79(s,3H),3.03(m,4H),2.44-2.45(m,4H),2.30(s,3H),2.23(s,3H).

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. The use of a compound of formula I or a salt thereof in the manufacture of a bruton's tyrosine kinase inhibitor or in the manufacture of a medicament for the treatment or prevention of a bruton's tyrosine kinase mediated disease:

in the formula,

r is hydrogen, C₁-C₃Lower alkyl, C₁-C₃Lower alkoxy, halogen (e.g., F, Cl,Br), amino, substituted amino;

x is N or CR⁵R⁶；

Y is C or O;

when the X is N, the N is N,is a double bond, and Y is C;

when X is CR₅R₆When the temperature of the water is higher than the set temperature,is a single bond, and Y is O; or when X is CR₅R₆When the temperature of the water is higher than the set temperature,is a double bond, and Y is C;

R⁷and R⁸Each independently is H, or C₁-C₆An alkyl group.

2. The compound of claim 1, wherein B is selected from the group consisting of:

R⁴selected from:

3. use according to claim 1 or 2, wherein the compound is of formula I-1 below:

in the formula,

R²selected from: hydrogen, halogen, optionally substituted C₁-C₆Alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acyloxy, optionallyOptionally substituted acylamino, optionally substituted C₁-C₆Alkyl, CN, sulfonic acid, aminosulfonyl, carbamoyl, carboxy, optionally substituted alkoxycarbonyl, optionally substituted phenyl, optionally substituted N-alkylpiperazino, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR_aR_bOptionally substituted pyridyl; r_aAnd R_bEach independently selected from alkyl and alkenyl;

X、Y、B、R¹、R³、R⁴and m is as defined in claim 1.

4. The use according to claim 3, wherein the compound is of formula II-1 below:

in the formula,

B、R²、R³、R⁴m and n are as defined in claim 3;

alternatively, the compound is represented by the following formula II-2:

in the formula,

B、R²、R⁴、R⁵、R⁶m and n are as defined in claim 3;

alternatively, the compound is represented by the following formula II-3:

in the formula,

B、R¹、R²、R³、R⁴m and n are as defined in claim 3And (4) limiting.

5. The use according to claim 4,

in the formula II-1, the compound represented by the formula,

b is selected from benzene ring or five-membered ring containing nitrogen;

m and n are as defined in claim 4;

in the formula II-2, the compound represented by the formula,

b is a benzene ring;

R⁴selected from: optionally substituted amido;

m and n are as defined in claim 4.

6. Use of a compound selected from the group consisting of:

7. use according to any one of claims 1 to 6, wherein the Bruton's tyrosine kinase mediated disease is cancer or and autoimmune disorders.

8. The use of claim 7, wherein the cancer is selected from the group consisting of: acute Lymphocytic Leukemia (ALL), Chronic Myeloid Leukemia (CML), Mantle Cell Lymphoma (MCL), large intestine cancer; the autoimmune disorder disease includes rheumatoid arthritis, anti-organ transplant rejection, anti-psoriasis, and lupus erythematosus.

9. A method of treating or preventing bruton's tyrosine kinase mediated disease comprising administering a compound of any one of claims 1-6 or a pharmaceutical composition comprising the compound to a subject in need thereof.

10. A compound of formula I or a pharmaceutically acceptable salt thereof:

in the formula,

X、Y、B、R¹、R³、R⁴and m is as defined in claim 1 or 2;

wherein,

and/or

R⁴Selected from the group consisting of:

11. the compound or salt according to claim 10, wherein the compound is represented by the following formula I-1:

in the formula,

X、Y、B、R¹、R³、R⁴and m is as defined in claim 10.

12. As in claimThe compound of claim 11, wherein R' is C₁-C₆Alkyl (preferably C)₁-C₃Alkyl group), C₁-C₆Haloalkyl (preferably C)₁-C₃Haloalkyl).

13. The compound of any one of claims 10-12, wherein R is³Selected from the group consisting of:

14. the compound of claim 13, wherein R is³Comprises the following steps: