CN106928216A

CN106928216A - Compound, Preparation Method And The Use with ERK kinase inhibiting activities

Info

Publication number: CN106928216A
Application number: CN201511031186.8A
Authority: CN
Inventors: 曹建华; 耿美玉; 黄敏; 江磊; 李磊; 唐帅; 冯家权; 杨晓彤; 丁健
Original assignee: Shanghai Institute of Materia Medica of CAS; Shanghai Haihe Pharmaceutical Co Ltd
Current assignee: Shanghai Institute of Materia Medica of CAS; Shanghai Haihe Pharmaceutical Co Ltd
Priority date: 2015-12-31
Filing date: 2015-12-31
Publication date: 2017-07-07
Also published as: WO2017114510A1; CN107922405A; CN107922405B

Abstract

The invention provides a kind of compound with ERK kinase inhibiting activities, Preparation Method And The Use, specifically, the invention provides compound of formula I, its stereoisomer, racemic modification or its pharmaceutically acceptable salt, and there is provided its application in the medicine for the prevention and treatment disease related to ERK kinases is prepared.

Description

Compound with ERK kinase inhibitory activity, preparation method and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a compound or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing the compound or the salt, which is used as a regulator of an ERK pathway or an inhibitor of ERK kinase, particularly ERK1 and ERK2 kinase.

Background

Extracellular signal-regulated kinases (ERKs) are a class of serine/threonine protein kinases found in the 90's of the 20 th century, and are one of the important subfamilies of the mitogen-activated protein kinase MAPKs family. Activated ERK can transmit extracellular signals to the nucleus, promote phosphorylation of cytoplasmic target proteins or regulate the activity of other protein kinases, thereby regulating gene expression. Its signaling is central to the signaling network involved in regulating cell growth, development and differentiation. Thus, ERK is involved in various biological effects of cell proliferation, differentiation, migration, invasion and apoptosis.

The Ras/Raf/MEK/ERK pathway is a main signal pathway related to the function of ERK, and node proteins on the pathway are hot spots for developing cancer-targeted drugs in recent years because the pathway regulates the proliferation, differentiation and apoptosis of cells. Specific B-Raf inhibitors Vemurafenib and dabrafenib are marketed in 2011 and 2013 respectively for the treatment of melanoma, wherein dabrafenib is used for treating B-RafV600E mutant non-small cell lung cancer and achieves breakthrough drug qualification of FDA. The MEK1/2 inhibitor trametinib was also marketed in 2013 for the treatment of melanoma. However, inhibition of these upstream pathway nodes has its limitations, tumors can rapidly develop drug resistance to B-Raf and MEK inhibitors, and the mechanism of drug resistance generation includes point mutation, change of protein polymeric form, change of protein peptide chain length, etc., which is a great obstacle for the next generation of drug resistance Raf and MEK drugs. ERK is used as a downstream key node of the pathway, no drug-resistant mutation is found at present, and the ERK targeting drug can greatly improve the treatment of patients with drug resistance to upstream target inhibitors, so that the ERK targeting drug is a promising anti-cancer drug research and development field.

In view of the above, there is an urgent need in the art to develop new ERK inhibitor drugs.

Disclosure of Invention

The invention aims to provide a compound which has a novel structure and can effectively inhibit ERK kinase, and a preparation method and application thereof.

In a first aspect of the invention, there is provided a compound of formula I, stereoisomers, racemates, or pharmaceutically acceptable salts thereof:

in the formula, X₁、X₂、X₃、X₄、X₅And X₆Each independently selected from CR₅Or N;

wherein R is₅Selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, -OH, cyano, halogen, amino, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted C1-C8 alkylcarbonyl, substituted or unsubstituted C1-C8 alkoxycarbonyl, substituted or unsubstituted C1-C8 carboxyl, substituted or unsubstituted C1-C8 ester, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

R₁selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, halogenated C1-C8 alkyl, halogenated C2-C8 alkenyl, halogenated C2-C8 alkynyl, substituted or unsubstituted 3-8 membered cyclic hydrocarbon, and substituted or unsubstituted aryl;

R₂selected from the group consisting of: substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted 3-to 8-membered cycloalkyl, and substituted or unsubstituted 3-to 8-membered heterocyclyl;

R₃selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, -OH, cyano, halogen, C1-C8 alkylenehydroxy, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

or, R₃And X₄And adjacent C and N atoms together form a substituted or unsubstituted 4-8 membered ring, wherein said ring contains at least 1N heteroatom and contains a total of 1-3 heteroatoms selected from O, S and N, and said ring is saturated or unsaturated;

R₄selected from substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, -CO (CR)₆R₇)_mR₈、-SO₂(CR₆R₇)_mR₈、-CONR₉(CR₆R₇)_mR₈、-COO(CR₆R₇)_mR₈Amino, C1-C8 carboxy;

m is 0, 1,2 or 3;

each R₆And R₇Each independently selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, and halogen, or R₆And R₇Joined to form a substituted or unsubstituted 3-to 6-membered ring;

each R₈Selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted 3-to 8-membered cycloalkyl, and substituted or unsubstituted 3-to 8-membered heterocyclyl;

each R₉Selected from the group consisting of: H. -OH, substituted or unsubstituted C1-C8 alkyl, C1-C8 alkylenehydroxy, and substituted or unsubstituted C1-C8 alkoxy.

In another preferred embodiment, the substituent means having one or more (e.g., 1 to 3) substituents selected from the group consisting of: halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkylhydroxy, -OH, C1-C3 alkoxy, C1-C3 alkylamino, 3-8 membered cyclic hydrocarbon, 3-8 membered heterocyclic group, amino and nitro.

In another preferred embodiment, R is₃And X₄And the linked "═ C-N-" or "— C-N-" together form a substituted or unsubstituted 4-8 membered ring.

In another preferred embodiment, R is₃And X₄And the linked "═ C-N-" or "— C-N-" together form a 4-8 membered ring including fused rings, spiro rings, or bridged rings.

In another preferred embodiment, the aryl group includes C5-C20 aryl and C3-C20 heteroaryl, wherein the heteroaryl contains 1-3 heteroatoms selected from the group consisting of: o, S and N.

In another preferred embodiment, the aryl group is selected from the group consisting of: phenyl, pyridyl, pyrazolyl, thiazolyl, imidazolyl, isoxazolyl, and oxazolyl.

In another preferred embodiment, X is₁、X₂、X₃、X₄、X₅、X₆Each independently selected from CR₅Or N; and, X₁、X₂、X₅At least one of which is N.

In another preferred embodiment, X is₁、X₂、X₅、X₆At least one or two of them are N, and the rest are C; and X₃、X₄Is C.

In another preferred embodiment, R₂Is a substituted or unsubstituted 5-6 membered saturated or unsaturated heterocyclic ring, wherein said substitution refers to having one or more (e.g., 1-3) substituents selected from the group consisting of: halogen, C1-C3 alkyl, -OH, amino, cyano, C1-C8 alkoxy, C1-C8 alkylamino.

In another preferred embodiment, R₂Is a substituted or unsubstituted 5-membered heterocyclic ring.

In another preferred embodiment, R₂Is a 5-membered heterocyclic ring containing 1-2N or a six-membered heterocyclic ring containing 1O.

In another preferred embodiment, R₂Is a 5-6 membered heterocyclic ring containing 1-3 substituents selected from the group consisting of: halogen, C1-C3 alkyl, -OH, amino, cyano, C1-C8 alkoxy, C1-C8 alkylamino.

In another preferred embodiment, R₄is-CO (CR)₆R₇)_mR₈And R is₈Is a substituted or unsubstituted aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group, wherein said substitution refers to having one or more (e.g., 1-3) substituents selected from the group consisting of: halogen, C1-C3 alkyl, -OH, amino, cyano, C1-C8 alkoxy, C1-C8 alkylamino.

In another preferred embodiment, the substitution is 1 to 3 substituents selected from the group consisting of: halogen, C1-C3 alkyl, and C1-C3 alkoxy.

In another preferred embodiment, R₈Is substituted or substituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted imidazolyl.

In another preferred embodiment, for R₃And X₄And a linked "═ C-N-" or "— C-N-" together form a substituted or unsubstituted 4-to 8-membered ring, preferably a 5-to 8-membered ring, more preferably a 5-, 6-, 7-or 8-membered ring containing 1 or 2N, or a 5-, 6-, 7-and 8-membered ring containing N and O.

In another preferred embodiment, R₄is-CO (CR)₆R₇)_mR₈Wherein R is₆And R₇Each independently selected from H or alkyl, substituted or unsubstituted C1-C8 alkylene hydroxyl, and m is 1 or 2, R₈Selected from H, substituted or unsubstituted pyridyl, substituted or unsubstituted phenyl.

In another preferred embodiment, the compound of formula I is represented by formula Ia:

wherein, X₁、X₂、X₃、X₄、X₅、X₆Each independently selected from CR₅Or N;

R₃selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, -OH, cyano, halogen, C1-C8 alkylenehydroxy, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted 3-8 membered aryl, and substituted or unsubstituted 3-8 membered heteroaryl;

R₂、R₄、R₅the definitions of (a) are the same as the previous definitions.

In another preferred embodiment, the compound of formula Ia is represented by the formula:

wherein R is₃Selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, -OH, cyano, halogen, C1-C8 alkylenehydroxy, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

In another preferred embodiment, R₂Selected from the group consisting of:

wherein each Ra is independently selected from substituted or unsubstituted C1-C4 alkyl; rb is selected from the group consisting of: halogen, -OH, cyano, amino, substituted or unsubstituted C1-C3 alkyl, C1-C3 haloalkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl; n is 0, 1,2 or 3;

R₃selected from the group consisting of H, substituted or unsubstituted C1-C8 alkyl, -OH, cyano, halogen, C1-C8 alkylenehydroxy, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

or, R₃And X₄And linked "═ C-N-" or "— C-N-" together form a substituted or unsubstituted 4-8 membered ring, wherein said ring contains at least 1N heteroatom and contains a total of 1 to 3 members selected from the group consisting of: o, S and N, and the ring is a saturated or unsaturated ring;

R₄selected from substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy、-CO(CR₆R₇)_mR₈、-SO₂(CR₆R₇)_mR₈、-CONR₉(CR₆R₇)_mR₈、-COO(CR₆R₇)_mR₈Amino, C1-C8 carboxy; wherein m is 0, 1,2 or 3;

each R₆、R₇Each independently selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, and halogen, or R₆And R₇Joined to form a substituted or unsubstituted 3-to 6-membered ring;

each R₉Selected from the group consisting of: H. -OH, substituted or unsubstituted C1-C8 alkyl, C1-C8 alkylenehydroxy, and substituted or unsubstituted C1-C8 alkoxy;

X₁、X₂、X₃、X₄、X₅and X₆Each independently selected from CR₅Or N;

R₅selected from the group consisting of H, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, -OH, cyano, halogen, amino, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted C1-C8 alkylcarbonyl, substituted or unsubstituted C1-C8 alkoxycarbonyl, substituted or unsubstituted C1-C8 carboxyl, substituted or unsubstituted C1-C8 ester, substituted or unsubstituted 3-8 membered cyclic hydrocarbon, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

In another preferred embodiment, R₅Selected from H or cyano.

In another preferred embodiment, the compound of formula I is represented by formula Ib below:

wherein,

X₁、X₂、X₃、X₅、X₆each independently selected from CR₅Or N;

p is 0, 1,2,3 or 4;

q is 1,2,3,4 or 5;

and p + q is less than or equal to 5;

y and Z are each independently selected from-CR^cR^d、O、S、-NR^c(ii) a Wherein R is^c、R^dEach independently selected from: H. substituted or unsubstituted C1-C8 alkyl, -OH, amino, halogen, cyano, substituted or unsubstituted C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted amino C1-C8 alkyl-, substituted or unsubstituted C1-C8 alkylamino, or-CR^cR^dis-C (═ O) -;

In another preferred embodiment, the compound of formula I is:

wherein,

p is 0, 1,2,3 or 4;

q is 1,2,3,4 or 5;

and p + q is less than or equal to 5;

y and Z are each independently selected from-CR^cR^d、O、S、-NR^c(ii) a Wherein R is^c、R^dEach independently selected from: H. substituted or unsubstituted C1-C8 alkyl, -OH, amino, halogen, cyano, substituted or unsubstituted C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted amino C1-C8 alkyl, substituted or unsubstituted C1-C8 alkylamino, or-CR^cR^dis-C (═ O);

In another preferred embodiment, in said compounds of formula Ia or Ib, R₂Selected from the group consisting of:

wherein each Ra is independently selected from: C1-C4 alkyl;

rb is selected from halogen, -OH, cyano, amino, substituted or unsubstituted C1-C3 alkyl, C1-C3 haloalkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl;

n is 0, 1,2 or 3;

p is 0, 1,2,3 or 4;

q is 1,2,3,4 or 5;

and p + q is less than or equal to 5;

y and Z are each independently selected from-CR^cR^d、O、S、-NR^c(ii) a Wherein R is^c、R^dEach independently selected from: H. substituted or unsubstituted C1-C8 alkyl, -OH, amino, halogen, cyano, C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, amino C1-C8 alkyl, substituted or unsubstituted C1-C8 alkylamino, or-CR^cR^dis-C (═ O);

R₄selected from substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, -CO (CR)₆R₇)_mR₈、-SO₂(CR₆R₇)_mR₈、-CONR₉(CR₆R₇)_mR₈、-COO(CR₆R₇)_mR₈Amino, carboxyl; wherein m is 0, 1,2 or 3;

each R₆、R₇Each independently selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, C1-C8 alkylenehydroxy, substituted or unsubstituted C1-C8 alkoxy, and halogen, or R₆And R₇Joined to form a substituted or unsubstituted 3-to 5-membered ring;

In another preferred embodiment, in the compound I, R is₁＝H，R₂Methyl-substituted five-membered heterocycles, R₄＝-CO(CR₆R₇)_mR₈Wherein R is₆＝R₇H, alkyl, alkylhydroxy, and m is 1 or 2, R₈Substituted or unsubstituted phenyl, pyridyl, or H.

In another preferred embodiment, said compound I is selected from the group consisting of:

in a second aspect of the invention, there is provided a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the group consisting of a compound according to the first aspect of the invention, stereoisomers, racemates thereof or pharmaceutically acceptable salts thereof and a pharmaceutically acceptable excipient.

In a third aspect of the present invention, there is provided a compound according to the first aspect of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the second aspect of the present invention for use in the preparation of a medicament for the prevention and treatment of diseases associated with ERK kinase and targeted inhibitors of ERK kinase.

In a fourth aspect of the invention, there is provided a process for the preparation of a compound according to the first aspect of the invention, comprising the steps of:

a) reacting (1e) with (1f) in an inert solvent under metal catalysis or acid/base catalysis to obtain a compound shown in the formula I;

wherein, X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃The groups are as defined in the first aspect of the invention;

LG₂is a leaving group selected from the group consisting of: halogen, sulfonate, methylthio, methyl sulfone.

In another preferred example, the method further comprises: steps (a-1) and (a-2), thereby producing a compound of formula (1 e):

(a-1) in an inert solvent, the (1a) and (1b) are subjected to condensation reaction or reductive amination reaction to obtain a compound (1 c);

(a-2) carrying out a coupling reaction of the (1c) and the compound (1d) in an inert solvent under the action of a metal catalyst to obtain a compound (1 e);

in the formula, LG₁Is a leaving group selected from the group consisting of: halogen, sulfonate, boric acid, borate, organotin, organozinc;

LG₂is a leaving group selected from the group consisting of: halogen, sulfonate, methylthio, methyl sulfone;

LG₃is a leaving group selected from the group consisting of: halogen, sulfonate, boric acid, borate;

FG is selected from the group consisting of: carboxylic acids, aldehydes, halogens;

X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃the definition of each group is as described in the first aspect of the invention.

In another preferred embodiment, in the (a-1), the reaction is carried out in an inert solvent selected from the group consisting of: water, methanol, ethanol, isopropanol, ethylene glycol, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, toluene, dichloromethane, chloroform, 1, 2-dichloroethane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, dioxane, or a combination thereof.

In another preferred embodiment, in the (a-1), the condensation reaction is carried out in the presence of a condensing agent selected from the group consisting of: 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, O-benzotriazol-tetramethyluronium hexafluorophosphate, or a combination thereof.

In another preferred embodiment, in the (a-1), the reductive amination reaction is carried out in the presence of a catalyst and a reducing agent, and the catalyst is selected from the group consisting of: titanium tetraisopropoxide, trifluoroacetic acid, acetic acid, formic acid, hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, or combinations thereof; the reducing agent is selected from the group consisting of: sodium borohydride, sodium cyanoborohydride, sodium borohydride acetate, sodium trifluoroacetyloxyborohydride, polymer-supported sodium borohydride reducing agent, sodium trimethoxyborohydride, sodium triethylborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, lithium borohydride, lithium aluminum hydride, or a combination thereof.

In another preferred embodiment, in the (a-2), the metal catalyst is selected from the group consisting of: tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) Tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Palladium acetate, palladium chloride, dichlorobis (triphenylphosphine) palladium, palladium trifluoroacetate, triphenylphosphine palladium acetate, [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride, bis (tri-o-phenylphosphino) palladium dichloride, 1, 2-bis (diphenylphosphino) ethane palladium dichloride, or a combination thereof.

In another preferred embodiment, in the (a), the reaction is carried out in the presence of a catalyst ligand selected from the group consisting of: tri-tert-butylphosphine, tri-tert-butylphosphine tetrafluoroborate, tri-n-butylphosphine, triphenylphosphine, tri-p-benzylphosphine, tricyclohexylphosphine tetrafluoroborate, tri-o-benzylphosphine, or combinations thereof.

In another preferred embodiment, in the (a), the reaction is carried out in the presence of a base, and the base includes an inorganic base and an organic base.

In another preferred embodiment, in the (a), the inorganic base is selected from the group consisting of: sodium hydride, potassium hydroxide, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, or a combination thereof.

In another preferred embodiment, in the (a), the organic base is selected from the group consisting of: pyridine, triethylamine, N-diisopropylethylamine, 1, 8-diazabicyclo [5.4.0] undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide, lutidine, or a combination thereof.

In another preferred embodiment, in the (a), the reaction is carried out in the presence of an acid selected from the group consisting of: hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, formic acid, acetic acid, or combinations thereof.

In another preferred embodiment, the temperature of step a) is-78 ℃ to 250 ℃.

In another preferred example, the step a) is performed under normal temperature conditions.

In another preferred embodiment, said step a) is carried out under dry ice bath or ice bath conditions.

In another preferred embodiment, said step a) is carried out under heating selected from the group consisting of: electrical heating, microwave heating, or a combination thereof.

In a fifth aspect of the invention, there is provided a process for the preparation of a compound according to the first aspect of the invention, comprising the steps of:

b) carrying out coupling reaction on the (1c) and the (1g) compound in an inert solvent under the catalysis of metal to prepare a compound shown in the formula I;

LG₁is a leaving group selected from the group consisting of: halogen, sulfonate, boric acid, borate, organotin, organozinc;

LG₃is a leaving group selected from the group consisting of: halogen, sulfonate, boric acid, borate.

In another preferred example, the method further comprises: step (b-1) and/or (b-2):

(b-1) (1a) and (1b) are coupled by condensation, reductive amination or the like in an inert solvent to obtain (1 c);

(b-2) (1d) and (1f) in an inert solvent in the presence of a base to give (1 g);

In another preferred embodiment, the (b-1) is carried out in an inert solvent selected from the group consisting of: water, methanol, ethanol, isopropanol, ethylene glycol, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, toluene, dichloromethane, chloroform, 1, 2-dichloroethane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, dioxane, or a combination thereof.

In another preferred embodiment, in the (b-1), the condensation reaction is carried out in the presence of a condensing agent selected from the group consisting of: 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, O-benzotriazol-tetramethyluronium hexafluorophosphate, the like, or combinations thereof.

In another preferred embodiment, in the (b-1), the reductive amination reaction is carried out in the presence of a catalyst and a reducing agent, the catalyst being selected from the group consisting of tetraisopropoxytitanium, trifluoroacetic acid, acetic acid, formic acid, hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, or a combination thereof; the reducing agent is selected from the group consisting of sodium borohydride, sodium cyanoborohydride, sodium acetate borohydride, sodium trifluoroacetyloxyborohydride, polymer-supported sodium borohydride reducing agent, sodium trimethoxyborohydride, sodium triethylborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, lithium borohydride, lithium aluminum tetrahydroborate, or combinations thereof.

In another preferred embodiment, in the (b), (1c) and (1g) are coupled in the presence of a metal catalyst selected from the group consisting of: tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) Tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Palladium acetate, palladium chloride, dichlorobis (triphenylphosphine) palladium, palladium trifluoroacetate, triphenylphosphine palladium acetate, [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride, bis (tri-o-phenylphosphino) palladium dichloride, 1, 2-bis (diphenylphosphino) ethane palladium dichloride, or a combination thereof.

In another preferred embodiment, in the (b), (1c) and (1g) are coupled in the presence of a metal catalyst ligand selected from the group consisting of: tri-tert-butylphosphine, tri-tert-butylphosphine tetrafluoroborate, tri-n-butylphosphine, triphenylphosphine, tri-p-benzylphosphine, tricyclohexylphosphine tetrafluoroborate, tri-o-benzylphosphine, or combinations thereof.

In another preferred embodiment, in the (b-2), (1d) and (1f) are coupled in the presence of a base, wherein the base comprises an inorganic base and an organic base.

In another preferred embodiment, in the (b-2), (1d) and (1f) are coupled in the presence of an inorganic base selected from the group consisting of: sodium hydroxide, lithium bistrimethylsilylamide, sodium bistrimethylsilylamide, potassium bistrimethylsilylamide, butyl lithium, lithium diisopropylamide, potassium hydroxide, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, or combinations thereof.

In another preferred embodiment, the coupling of (1d) and (1f) in (b-2) is carried out in the presence of an organic base selected from the group consisting of: pyridine, triethylamine, N, N-diisopropylethylamine, 1, 8-diazabicyclo [5.4.0] undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide, lutidine, or a combination thereof.

In another preferred embodiment, in the (b-2), (1d) and (1f) are coupled in the presence of an acid selected from the group consisting of: hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, formic acid, acetic acid, or combinations thereof.

In another preferred embodiment, the temperature of step b) is-78 ℃ to 250 ℃.

In another preferred example, the step b) is performed under normal temperature conditions.

In another preferred embodiment, said step b) is carried out under dry ice bath or ice bath conditions.

In another preferred embodiment, said step b) is carried out under heating selected from the group consisting of: electrical heating, microwave heating, or a combination thereof.

In a sixth aspect of the invention, there is provided a method of non-therapeutically inhibiting ERK kinase activity comprising the steps of: contacting a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, with ERK kinase, thereby inhibiting ERK kinase.

In another preferred embodiment, the contacting is contacting purified ERK kinase or cells expressing ERK kinase.

In a seventh aspect of the invention, there is provided a method for the prevention and/or treatment of a disease associated with ERK kinase activity in a mammal, comprising administering to a mammal in need thereof a therapeutically effective amount of a compound according to the first aspect of the invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or administering a therapeutically effective amount of a pharmaceutical composition according to the second aspect of the invention.

In another preferred embodiment, the ERK kinase comprises ERK1, ERK2, or a combination.

In another preferred embodiment, the diseases related to ERK kinase activity refer to diseases related to high expression or high activity of ERK kinase.

In another preferred embodiment, the disease associated with ERK kinase activity is selected from the group consisting of: a tumor.

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 inventor of the invention unexpectedly finds that a compound shown as a formula I or a pharmaceutically acceptable salt thereof can be used as an ERK kinase inhibitor and has high inhibitory activity for the first time through extensive and intensive research. The present invention has been completed based on this finding.

Description of the terms

As used herein, the term "about" when used in reference to a specifically recited value means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

Unless defined otherwise, the following terms used in the specification and claims have the meanings that are commonly understood by those skilled in the art. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter claimed. In this application, the use of the singular also includes the plural unless specifically stated otherwise. It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the terms "comprising" or "including" can be open, semi-closed, and closed. In other words, the term also includes "consisting essentially of …," or "consisting of ….

Definitions for the terms of the standardization industry can be found in the literature references including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4. THED." vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise indicated, conventional methods within the skill of the art are employed, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods. Unless a specific definition is set forth, the terms used herein in the pertinent description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the instructions of the kit from the manufacturer, or according to the methods known in the art or the instructions of the present invention. The techniques and methods described above can generally be practiced according to conventional methods well known in the art, as described in various general and more specific documents referred to and discussed in this specification. In the present specification, groups and substituents thereof may be selected by one skilled in the art to provide stable moieties and compounds.

When a substituent is described by a general formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the formula is written from right to left. For example, -CH₂O-is equivalent to-OCH₂-。

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and treatises, are hereby incorporated by reference in their entirety.

Certain chemical groups defined herein are preceded by a shorthand notation to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the shorthand notation excludes carbons that may be present in a substituent of the group.

In addition to the foregoing, the following terms, when used in the specification and claims of this application, have the meanings indicated below, unless otherwise specifically indicated.

In the present application, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

"hydroxy" means an-OH group.

"hydroxyalkyl" refers to an alkyl group as defined below substituted with a hydroxyl group (-OH).

"carbonyl" refers to a-C (═ O) -group.

"nitro" means-NO₂。

"cyano" means-CN.

"amino" means-NH₂。

"substituted amino" refers to an amino group substituted with one or two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl groups as defined below, e.g., monoalkylamino, dialkylamino, alkylamido, aralkylamino, heteroaralkylamino.

"carboxyl" means-COOH.

In this application, the term "alkyl" as a group or as part of another group (e.g., as used in halo-substituted alkyl and the like groups) refers to a fully saturated straight or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms, having, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms, and attached to the remainder of the molecule by a single bond, including, for example, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, decyl and the like. For the purposes of the present invention, the term "alkyl" refers to alkyl groups containing from 1 to 6 carbon atoms.

In the present application, the term "alkenyl" as a group or part of another group means a straight or branched hydrocarbon chain group consisting of only carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms, and being connected to the rest of the molecule by a single bond, such as, but not limited to, vinyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1, 4-dienyl, and the like.

In the present application, the term "alkynyl" as a group or part of another group refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, optionally containing at least one double bond, having for example 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and being connected to the rest of the molecule by single bonds, such as but not limited to ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-en-4-ynyl and the like.

In the present application, the term "cycloalkyl" as a group or as part of another group means a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting of only carbon and hydrogen atoms, which may include fused, bridged or spiro ring systems, having from 3 to 15 carbon atoms, preferably from 3 to 10 carbon atoms, more preferably from 3 to 8 carbon atoms, and which is saturated or unsaturated and may be attached to the rest of the molecule by a single bond via any suitable carbon atom. Unless otherwise specifically indicated in the specification, carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2, 3-indanyl, 1,2,3, 4-tetrahydro-naphthyl, 5,6,7, 8-tetrahydro-naphthyl, 8, 9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8, 9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9, 10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo [2.2.1] heptyl, 7-dimethyl-bicyclo [2.2.1] heptyl, bicyclo [2.2.1] heptenyl, bicyclo [2.2.2] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octenyl, Bicyclo [3.2.1] octenyl, adamantyl, octahydro-4, 7-methylene-1H-indenyl, octahydro-2, 5-methylene-pentalenyl and the like.

In this application, the term "heterocyclyl" as a group or part of another group means a stable 3-to 20-membered non-aromatic cyclic group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur. Unless otherwise specifically indicated in the specification, a heterocyclic group may be a monocyclic, bicyclic, tricyclic or higher ring system, which may include fused ring systems, bridged ring systems or spiro ring systems; wherein the nitrogen, carbon or sulfur atom in the heterocyclic group may be optionally oxidized; the nitrogen atoms may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule via a carbon atom or a heteroatom and by a single bond. In heterocyclic groups containing fused rings, one or more of the rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, heterocyclyl is preferably a stable 4-to 11-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 4-to 8-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2, 7-diaza-spiro [3.5] nonan-7-yl, 2-oxa-6-aza-spiro [3.3] heptan-6-yl, 2, 5-diaza-bicyclo [2.2.1] heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolanyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl, phthalimidyl, and the like.

In this application, the term "aryl" as a group or as part of another group means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. For the purposes of the present invention, an aryl group may be a monocyclic, bicyclic, tricyclic or higher polycyclic ring system and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the aryl group is attached to the remainder of the molecule by a single bond via an atom on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl, 2, 3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazin-3 (4H) -one-7-yl, and the like.

In the present application, the term "arylalkyl" refers to an alkyl group as defined above substituted with an aryl group as defined above.

In this application, the term "heteroaryl" as a group or part of another group means a 5-to 16-membered conjugated ring system group having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur in the ring. Unless otherwise specifically indicated in the specification, a heteroaryl group may be a monocyclic, bicyclic, tricyclic or higher ring system, and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is attached to the rest of the molecule by a single bond via an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group may be optionally oxidized; the nitrogen atoms may optionally be quaternized. For the purposes of the present invention, heteroaryl is preferably a stable 5-to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 5-to 10-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur or a 5-to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindolyl, purinyl, quinolyl, isoquinolyl, diazonaphthyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxazolyl, cinnolinyl, quinazolinyl, thiophenyl, indolizinyl, orthophenanthrolidinyl, isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6, 7-tetrahydrobenzo [ b ] thienyl, naphthopyridyl, pyridinyl, and the like, [1,2,4] triazolo [4,3-b ] pyridazine, [1,2,4] triazolo [4,3-a ] pyrazine, [1,2,4] triazolo [4,3-c ] pyrimidine, [1,2,4] triazolo [4,3-a ] pyridine, imidazo [1,2-b ] pyridazine, imidazo [1,2-a ] pyrazine and the like.

In the present application, the term "heteroarylalkyl" refers to an alkyl group as defined above substituted with a heteroaryl group as defined above.

In this application, "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups. The "optionally" substituents described in the claims and the description section of the present invention are selected from alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, cyano, nitro, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl.

The terms "moiety," "structural moiety," "chemical moiety," "group," "chemical group" as used herein refer to a specific fragment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities that are embedded in or attached to a molecule.

"stereoisomers" refers to compounds that consist of the same atoms, are bonded by the same bonds, but have different three-dimensional structures. The present invention is intended to cover various stereoisomers and mixtures thereof.

When the compounds of the present invention contain olefinic double bonds, the compounds of the present invention are intended to include both E-and Z-geometric isomers unless otherwise specified.

"tautomer" refers to an isomer formed by the transfer of a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the invention are also intended to be included within the scope of the invention.

The compounds of the present invention or pharmaceutically acceptable salts thereof may contain one or more chiral carbon atoms and may therefore give rise to enantiomers, diastereomers and other stereoisomeric forms. Each chiral carbon atom may be defined as (R) -or (S) -, based on stereochemistry. The present invention is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the invention may be prepared by selecting as starting materials or intermediates racemates, diastereomers or enantiomers. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, e.g., crystallization and chiral chromatography.

Conventional techniques for preparing/separating individual isomers include Chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, Chiral high performance liquid chromatography, as described, for example, in Gerald Gubitz and Martin G.Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol.243, 2004; m. Stalcup, Chiral Separations, Annu. Rev. anal. chem.3:341-63, 2010; fumiss et al (eds.), VOGEL' S ENCYCOPEDIA OFPRACTICAL ORGANIC CHEMISTRY 5. TH ED., Longman Scientific and technical Ltd., Essex,1991, 809-816; heller, acc, chem, res, 1990,23,128.

In the present application, the term "pharmaceutically acceptable salts" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"pharmaceutically acceptable acid addition salts" refers to salts with inorganic or organic acids which retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, and the like; organic acid salts include, but are not limited to, formates, acetates, 2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, caprates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbates, salicylates, 4-aminosalicylates, napadisylates, and the like. These salts can be prepared by methods known in the art.

"pharmaceutically acceptable base addition salts" refers to salts with inorganic or organic bases which maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

"polymorph" refers to different solid crystalline phases of certain compounds of the present invention in the solid state due to the presence of two or more different molecular arrangements. Certain compounds of the present invention may exist in more than one crystalline form and the present invention is intended to include the various crystalline forms and mixtures thereof.

Typically, crystallization will result in solvates of the compounds of the invention. The term "solvate" as used herein refers to an aggregate comprising one or more molecules of the compound of the present invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the compounds of the present invention may exist as hydrates, including monohydrates, dihydrate, hemihydrate, sesquihydrates, trihydrate, tetrahydrate, and the like, as well as the corresponding solvated forms. The compounds of the invention may form true solvates, but in some cases it is also possible to retain only adventitious water or a mixture of water plus a portion of adventitious solvent. The compounds of the invention may be reacted in a solvent or precipitated or crystallized from a solvent. Solvates of the compounds of the invention are also included within the scope of the invention.

The invention also includes prodrugs of the above compounds. In the present application, the term "prodrug" denotes a compound that can be converted under physiological conditions or by solvolysis to the biologically active compound of the invention. Thus, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of a compound of the invention. Prodrugs may not be active when administered to a subject in need thereof, but are converted in vivo to the active compounds of the invention. Prodrugs are generally rapidly converted in vivo to yield the parent compound of the invention, for example, by hydrolysis in blood. Prodrug compounds generally provide solubility, histocompatibility, or sustained release advantages in mammalian organisms. Prodrugs include known amino protecting groups and carboxyl protecting groups. Specific methods for preparing prodrugs can be found in Saulnier, M.G., et al, bioorg.Med.chem.Lett.1994,4, 1985-1990; greenwald, r.b., et al, j.med.chem.2000,43,475.

In the present application, a "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for delivery of biologically active compounds to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of active ingredients and exert biological activity.

The term "pharmaceutically acceptable" as used herein refers to a substance (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., the substance can be administered to an individual without causing an adverse biological response or interacting in an adverse manner with any of the components contained in the composition.

As used herein, "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent, or emulsifying agent that is approved by the relevant governmental regulatory agency for human or livestock use.

The "tumor" and "diseases related to abnormal cell proliferation" include, but are not limited to, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, squamous cell lung cancer, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, renal cancer, oral cancer, and the like.

The terms "preventing," "prevention," and "prevention" as used herein include reducing the likelihood of occurrence or worsening of a disease or disorder in a patient.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) preventing the occurrence of a disease or condition in a mammal, particularly when such mammal is susceptible to the disease or condition, but has not been diagnosed as having the disease or condition;

(ii) inhibiting the disease or disorder, i.e., arresting its development;

(iii) alleviating the disease or condition, i.e., causing regression of the state of the disease or condition; or

(iv) Alleviating the symptoms caused by the disease or disorder.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The terms "administering," "administration," "administering," and the like as used herein refer to a method capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Administration techniques useful for the compounds and methods described herein are well known to those skilled in the art, for example, in Goodman and Gilman, the pharmacological Basis of Therapeutics, current ed.; pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

The terms "drug combination", "administering other treatment", "administering other therapeutic agent" and the like as used herein refer to a drug treatment obtained by mixing or combining more than one active ingredient, including fixed and unfixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one co-agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, concomitant administration, or sequential administration at variable intervals of at least one compound described herein and at least one synergistic formulation to a patient as separate entities. These also apply to cocktail therapy, for example the administration of three or more active ingredients.

It will also be appreciated by those skilled in the art that in the processes described below, the functional groups of the intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl groups (e.g.tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino include t-butyloxycarbonyl, benzyloxycarbonyl and the like. Suitable thiol protecting groups include-C (O) -R "(where R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable carboxyl protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups may be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting Groups is described in detail in Greene, T.W. and P.G.M.Wuts, Protective Groups in organic Synthesis, (1999),4th Ed., Wiley. The protecting group may also be a polymeric resin.

Preparation of Compounds of formula I

The following reaction schemes illustrate methods for preparing compounds of formula I, stereoisomers or mixtures thereof, or pharmaceutically acceptable salts thereof:

wherein,

X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃、R₄are as described above in the section for the embodiments of the compounds of formula I. It is understood that in the following reaction schemes, combinations of substituents and/or variables in the general formulae are permissible only if such combinations result in stable compounds. It will also be appreciated that other general formulae, such as general formula (Ia), (Ia-1), (Ia-2), (Ia-3), (Ia-4), (Ib-1), (Ib-2), (Ib-3), (Ib-4), and other compounds of formula I specifically disclosed herein, may be prepared by methods disclosed herein (by applying appropriately substituted starting materials and modifying the synthesis parameters as required using methods well known to those skilled in the art) or known methods by those skilled in the art of organic chemistry.

The skilled person will appreciate that in some cases the starting materials and intermediates in the preparation of the compounds of the invention may contain functional groups which need to be protected during the synthesis. The exact nature of any protecting group used will depend on the identity of the functional group being protected, as will be apparent to those skilled in the art. Guidance in the selection of suitable protecting Groups and synthetic strategies for their attachment and removal can be found, for example, in Green & Wuts, Green' sProtective Groups in organic Synthesis, ("protecting Groups in organic Synthesis") 3d Edition, Jon Wiley & Sons, Inc., New York (1999) and the references cited therein.

Thus, a protecting group refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces, or prevents the reactivity of that functional group. Generally, the protecting group can be selectively removed as desired during the synthesis.

Reaction scheme 1:

in each formula, X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃、R₄、LG₁And LG₂Are as described above in the section for the embodiments of the compounds of formula I.

Reaction scheme 2:

X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃、R₄、LG₁、LG₂and LG₃Are as described above in the section for the embodiments of the compounds of formula I.

The main advantages of the invention are:

1. provides a compound shown as a formula I.

2. Provides an ERK kinase inhibitor with novel structure, a preparation method and application thereof, wherein the inhibitor has higher inhibitory activity to ERK kinase.

3. Pharmaceutical compositions for treating diseases associated with ERK kinase activity are provided.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit 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 percentages and parts by weight.

The test materials and reagents used in the following examples are commercially available without specific reference.

Example 1

Synthesis of 1- (5-bromoindolin-1-yl) -2-phenyl ethanone

In a dry 50mL three-necked flask, compound 1(1.10g, 5.55mmol), phenylacetyl chloride (858mg, 5.55mmol), and triethylamine (1.68g, 16.66mmol) were sequentially added, and dissolved in dichloromethane (20 mL). Upon completion of the reaction by LCMS, concentrated directly under reduced pressure and purified by silica gel column (ethyl acetate: petroleum ether: 1: 10) to give product 2(1.2g, white solid) in yield: 68 percent.

LCMS:m/z 318.1(M+H)；RT＝1.40min(2min).

Synthesis of 2-phenyl-1- (5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) indolin-1-yl) ethanone

To a dry 50mL three-necked flask were added compound 2(1.4g, 4.42mmol), pinacol diboron (2.25g, 8.84mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (322mg, 0.44mmol), potassium acetate (866mg, 8.84mmol), 1, 4-dioxane (20mL) in that order. Heating to 100 ℃ under the protection of nitrogen and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed with saturated brine (50mL) in this order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and subjected to silica gel column (ethyl acetate: petroleum ether ═ 1: 20) to obtain product 3(1.3g, yellow solid) in yield: 94 percent.

LCMS:m/z 364.4(M+H)；RT＝1.53min(2min).

Synthesis of 1- (5- (2-chloropyrimidin-4-yl) indolin-1-yl) -2-phenylethanone

In a dry 50mL three-necked flask, compound 3(700mg, 1.92mmol), 2, 4-dichloropyrimidine (286mg, 1.92mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (139mg, 0.19mmol), potassium carbonate (400mg, 2.89mmol), 1, 4-dioxane (8mL) and water (2mL) were added in that order. Heating to 100 ℃ under the protection of nitrogen, and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure using a thick preparative plate (ethyl acetate: petroleum ether ═ 1: 5) to give product 5(250mg, yellow solid) in yield: 37 percent.

LCMS:m/z 350.1(M+H)；RT＝1.48min(2min).

Synthesis of 1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) -2-phenylethanone

Compound 4(200mg, 0.57mmol), 1-methyl-5-aminopyrazole (55mg, 0.57mmol), tris (dibenzylideneacetone) dipalladium (55mg, 0.06mmol), 4, 5-bis-diphenylphosphino-9, 9-dimethylxanthene (35mg, 0.06mmol), cesium carbonate (279mg,0.86mmol)1, 4-dioxane (10mL) were added in this order to a dry 50mL three-necked flask. Heating to 100 ℃ under the protection of nitrogen, and reacting for 4 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the product HE153(16mg, yellow solid) by reverse phase preparative column, yield: 7 percent.

LCMS:m/z 411.4(M+H)；RT＝1.27min(2min).

1H-NMR(MeOD 400MHz):8.38-8.39(m,1H),8.19-8.21(m,1H),7.96-7.98(m,2H),7.56(s,1H),7.26-7.38(m,7H),4.19-4.23(m,2H),3.89(s,2H),3.79(s,3H),3.20-3.23(m,2H).

Example 2

Synthesis of 1- (5-bromo-2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) -2-phenylethanone

Compound 5(398mg,2.0mmol), phenylacetic acid (272mg,2.0mmol), HATU (1.1g,3.0mmol) and DMF (10mL) were added successively to a dry 50mL one-neck flask, and N, N-diisopropylethylamine (517mg,4.0mmol) was added dropwise and reacted at room temperature overnight. After completion of the reaction, 15ml of water was added, extracted with ethyl acetate (20 ml. times.2), and the organic phases were combined. The organic phase was washed with saturated brine (15ml × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 6:1) to give compound 6(539mg, yield: 85%) as a yellow solid.

LCMS:m/z 318.8(M+H)⁺；RT＝1.652min(254nm)。

Synthesis of 1- (2-phenylacetyl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-5-ylboronic acid

In a dry 50mL three-necked flask, compound 7(317mg,1.0mmol),1, 4-dioxane (8mL), bis pinaboronate (381mg,1.5mmol), potassium acetate (194mg,2.0mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (73mg,0.1mmol) were added. The reaction was stirred for 6 hours at 90 ℃ under nitrogen atmosphere for 3 times under vacuum, and the reaction was completed by TLC, the reaction solution was cooled to room temperature, and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane: methanol 5:1) to obtain compound 8(169mg, yield: 60%) as a yellow solid.

LCMS:m/z 282.9(M+H)⁺。

Synthesis of 1- (5- (2-chloropyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) -2-phenylethanone

In a dry 50mL three-necked flask, compound 9(169mg,0.6mmol),2, 4-dichloropyrimidine (134mg, 0.9mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (44mg, 0.06mmol), cesium carbonate (391mg, 1.2mmol), 1, 4-dioxane (8mL) and water (0.5mL) were added. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 2:1) to give compound 10(105mg, yield: 50%) as a yellow solid.

LCMS:m/z 350.9(M+H)⁺。

Synthesis of 1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) -2-phenylethane-1-one

In a dry 50mL three-necked flask, compound 10(70mg, 0.2mmol), 1-methyl-5-aminopyrazole (29mg, 0.3mmol), tris (dibenzylideneacetone) dipalladium (18mg, 0.02mmol), 4, 5-bis-diphenylphosphino-9, 9-dimethylxanthene (10mg, 0.02mmol), cesium carbonate (130mg,0.4mmol) and 1, 4-dioxane (6mL) were added in this order. The mixture was heated to 105 ℃ under nitrogen and reacted overnight. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (15mL), washed with saturated brine (10mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to purify the crude product by silica gel column chromatography (petroleum ether: ethyl acetate ═ 1:2) to obtain compound a2(5mg, yield: 6%) as a yellow solid.

¹HNMR(400MHz,CDCl₃-d)8.81(s,1H)，8.46(d,1H,J＝5.2Hz)，8.09(s,1H)，7.51(d,1H,J＝2.0Hz)，7.39(d,2H,J＝7.2Hz)，7.32-7.17(m,4H)，6.89(s,1H)，6.35(d,1H,J＝2.0Hz)，4.61(s,2H)，4.18(t,2H,J＝8.8Hz)，3.82(s,3H)，3.12(t,2H,J＝8.4Hz)。

LCMS:m/z 412.1(M+H)⁺；RT＝1.179min(254nm)。

Example 3

Synthesis of N- (5-bromopyridin-2-yl) -2-phenylacetamide

Compound 11(2.00g,14.71mmol), 2-amino-5-bromopyridine (2.54g,14.71mmol) and HATU (5.59g,14.71mmol) were sequentially added to a dry 50mL single-necked flask in DMF (20mL), and N, N-diisopropylethylamine (1.91g,14.71mmol) was added dropwise and the mixture was reacted at room temperature for 4 hours. After LCMS detection, water was added to the reaction mixture, followed by extraction with ethyl acetate, washing of the organic phase with brine, drying over anhydrous sodium sulfate, filtration, and concentration, followed by silica gel column (developing solvent ethyl acetate: petroleum ether: 1: 10) to obtain compound 2(2.8g, white solid) in yield: 67%.

LCMS:m/z291.1(M+H)；RT＝1.33min(2.0min).

Synthesis of (6- (2-phenylacetylamino) pyridin-3-yl) boronic acid

In a dry 50mL three-necked flask were added compound 12(1.4g, 4.81mmol), pinacol diboron (2.44g, 9.62mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (351mg, 0.48mmol), potassium acetate (942mg, 9.62mmol), 1, 4-dioxane (20mL) in that order. Heating to 100 ℃ under the protection of nitrogen and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed with saturated brine (50mL × 1) in this order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and subjected to silica gel column (ethyl acetate: petroleum ether ═ 1: 3) to obtain product 13(1.0g, yellow solid) in yield: 81 percent.

LCMS:m/z257.1(M+H)；RT＝0.39min(2min).

Synthesis of N- (5- (2-chloropyrimidin-4-yl) pyridin-2-yl) -2-phenylacetamide

In a dry 50mL three-necked flask were added compound 13(1.0g, 3.91mmol), 2, 4-dichloropyrimidine (582mg, 3.91mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (293mg, 0.40mmol), potassium carbonate (810mg, 5.87mmol), 1, 4-dioxane (20mL) and water (5mL) in that order. Heating to 100 ℃ under the protection of nitrogen, and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a post-use plate (ethyl acetate: petroleum ether ═ 1: 3) to obtain the product 14(500mg, yellow solid) in yield: 40 percent.

LCMS:m/z325.2(M+H)；RT＝1.28min(2min).

Synthesis of N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) -2-phenylacetamide

Compound 14(50mg, 0.15mmol), 1-methyl-5-aminopyrazole (15mg, 0.15mmol), tris (dibenzylideneacetone) dipalladium (18mg, 0.02mmol), 4, 5-bis-diphenylphosphino-9, 9-dimethylxanthene (12mg, 0.02mmol), cesium carbonate (49mg,0.15mmol)1, 4-dioxane (10mL) were added sequentially in a dry 50mL three-necked flask. Heating to 100 ℃ under the protection of nitrogen, and reacting for 2 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the product a3(16mg, yellow solid) in reverse phase: 28 percent.

LCMS:m/z386.2(M+H)；RT＝1.26min(2min).

¹H-NMR(MeOD 400MHz):9.01-9.02(m,1H),8.46-8.47(m,2H),8.15-8.16(m,1H),7.52-7.53(m,1H),7.33-7.40(m,6H),6.42-6.43(m,1H),3.78-3.79(m,5H).

Example 4

Synthesis of 2-phenyl-N- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) acetamide

Compound 15(500mg,2.28mmol), phenylacetic acid (310mg,2.28mmol) and HATU (1.30g,3.42mmol) were sequentially added to a dry 50mL single-necked flask, and then dissolved in DMF (10mL) and triethylamine (461mg,4.56mmol) was added dropwise, followed by reaction at room temperature for 3 hours. After completion of LCMS detection, water was added to the reaction solution, followed by extraction with ethyl acetate, washing of the organic phase with saturated brine, drying over anhydrous sodium sulfate, filtration, and concentration, followed by silica gel column (developing solvent ethyl acetate: petroleum ether: 1: 20) to obtain compound 16(520mg, white solid) in yield: 68 percent.

LCMS:m/z338.2(M+H)；RT＝1.09min(2.0min).

Synthesis of N- (5- (2-chloropyrimidin-4-yl) pyridin-2-yl) -2-phenylethyl

In a dry 50mL three-necked flask were added compound 16(520mg, 1.54mmol), 2, 4-dichloropyrimidine (275mg, 1.85mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (110mg, 0.15mmol), potassium carbonate (319mg, 2.31mmol), 1, 4-dioxane (8mL) and water (2mL) in that order. Heating to 100 ℃ under the protection of nitrogen, and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a post-use plate (ethyl acetate: petroleum ether ═ 1: 5) to obtain the product 17(480mg, yellow solid) in yield: 96 percent.

LCMS:m/z324.3(M+H)；RT＝1.10min(2min).

Synthesis of N- (4- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) phenyl) -2-phenylacetamide

Compound 17(200mg, 0.62mmol), 1-methyl-5-aminopyrazole (60mg, 0.62mmol), tris (dibenzylideneacetone) dipalladium (155mg, 0.06mmol), 4, 5-bis-diphenylphosphino-9, 9-dimethylxanthene (35mg, 0.06mmol), cesium carbonate (302mg,0.93mmol)1, 4-dioxane (10mL) were added in this order to a dry 50mL three-necked flask. Heating to 100 ℃ under the protection of nitrogen, and reacting for 2 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed successively with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the product a4(50mg, yellow solid) in reverse phase preparation column, yield: 21 percent.

LCMS:m/z385.4(M+H)；RT＝1.13min(2min).

¹H-NMR(CDCl3,400MHz):8.22-8.24(m,1H),8.00(d,j＝8.4,2H),7.62(d,j＝8.4,2H),7.50-7.54(m,2H),7.34-7.44(m,6H),6.47(s,1H),3.79-3.88(m,5H).

Example 5

Synthesis of 4-bromo-2-fluoroaniline

In a 100mL round-bottom flask was added 4-bromo-2-fluoro-1-nitrobenzene (2.2g,10mmol), iron powder (2.8g,50mmol) and tetrahydrofuran (20mL), and hydrochloric acid (30mL, 2N) was added dropwise with stirring at room temperature, and stirred at room temperature for 2h, and anhydrous sodium carbonate (2g) and anhydrous sodium sulfate were added, followed by filtration, washing with ethyl acetate, and concentration under reduced pressure to give compound 19 as a yellow solid (1.65g, yield: 87%).

LCMS:m/z 191.1(M+H)⁺；RT＝1.405min。

N- (4-bromo-2-fluorophenyl) -2-phenylacetamide

In a dry 50mL one-neck flask were added compound 19(950mg,5mmol), phenylacetic acid (680mg,5mmol), HATU (3.42g,9mmol), DMF (15mL) and N, N-diisopropylethylamine (1.29g,10mmol) in that order. After stirring at room temperature for 6 hours, the mixture was diluted with ethyl acetate (20mL), washed with saturated brine (10mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 8:1) to give compound 20(1.07g, yield: 70%) as a yellow solid.

LCMS:m/z 309.8(M+H)⁺；RT＝1.457min。

Synthesis of N- (2-fluoro-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) -2-phenylacetamide

In a dry 50mL three-necked flask was added compound 20(1.01g,3.3mmol),1, 4-dioxane (10mL), bis pinaboronate (4.2g,16.7mmol), potassium acetate (648mg,6.6mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (220mg,0.3 mmol). The reaction mixture was cooled to room temperature, and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain compound 21(703mg, yield: 60%) as a yellow solid.

LCMS:m/z 355.8(M+H)⁺；RT＝1.699min。

Synthesis of N- (4- (2-chloropyrimidin-4-yl) -2-fluorophenyl) -2-phenylacetamide

In a dry 10mL round bottom flask was added, in order at room temperature, 21(355mg,1.0mmol), 2, 4-dichloropyrimidine (222mg, 1.5mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (73mg,0.1mmol), cesium carbonate (652mg, 2.0mmol), 1, 4-dioxane (6mL) and water (1 mL). The reaction mixture was cooled to room temperature, and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 3:1) to give compound 22(174mg, yield: 51%) as a yellow solid.

LCMS:m/z 341.9(M+H)⁺；RT＝1.677min。

Synthesis of N- (2-fluoro-4- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) phenyl) -2-phenylacetamide

In a dry 25mL three-necked flask, compound 22(68mg, 0.2mmol), 1-methyl-5-aminopyrazole (29mg, 0.3mmol), tris (dibenzylideneacetone) dipalladium (18mg, 0.02mmol), 4, 5-bis-diphenylphosphino-9, 9-dimethylxanthene (12mg, 0.02mmol), cesium carbonate (130mg,0.4mmol) and 1, 4-dioxane (3mL) were added in this order. And (3) heating to 100 ℃ by microwave under the protection of nitrogen, and reacting for 2 hours. After completion of the reaction, it was diluted with ethyl acetate (20mL), washed with saturated brine (10 mL. times.3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by acidic prep-HPLC to give Compound A5(6mg, yield: 7%) as a yellow solid.

¹HNMR(400MHz,CDCl₃-d)8.49(t,1H,J＝8.4Hz)，8.41(d,1H,J＝5.2Hz)，7.78(s,1H)，7.76(d,1H,J＝4.8Hz)，7.50-7.35(m,7H),7.16(d,1H,J＝5.6Hz)，6.35(d,1H,J＝1.6Hz)，3.81(s,5H)。

LCMS:m/z 402.9(M+H)⁺；RT＝1.385min。

The present invention, using the above analogous method, can produce the following compounds:

example 6

Synthesis of tert-butyl-5- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) indoline-1-carboxylic acid tert-butyl ester

¹HNMR(400MHz,CDCl₃-d)8.38(d,1H,J＝5.2Hz)，7.85(s,2H)，7.49(d,1H,J＝1.6Hz)，7.15(d,1H,J＝5.2Hz)，6.87(s,1H)，6.35(d,1H,J＝1.2Hz)，4.03(t,2H,J＝8.8Hz)，3.80(s,3H)，3.15(t,2H,J＝8.8Hz)，1.58(s,9H)。

LCMS:m/z 393.3(M+H)⁺；RT＝1.461min(254nm)。

Example 7

Synthesis of 1- (5- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) indolin-1-yl) -3-phenylpropan-1-one

¹HNMR(400MHz,CDCl₃-d)8.40(d,1H,J＝5.2Hz)，8.32(d,1H,J＝8.4Hz)，7.88-7.86(m,2H)，7.49(d,1H,J＝1.6Hz)，7.32-7.16(m,6H)，6.95(s,1H)，6.35(d,1H,J＝1.6Hz)，4.03(t,2H,J＝8.4Hz)，3.80(s,3H)，3.21(t,2H,J＝8.4Hz)，3.08(t,2H,J＝7.6Hz)，2.76(t,2H,J＝7.6Hz)。

LCMS:m/z 425.0(M+H)⁺；RT＝1.339min(254nm)。

Example 8

Synthesis of 4- (indolin-5-yl) -N- (1-methyl-1H-pyrazol-5-yl) pyrimidin-2-amine

¹HNMR(400MHz,DMSO-d₆)8.38(d,1H,J＝5.6Hz)，7.93-7.91(m,3H)，7.47(d,1H,J＝1.6Hz)，7.40(d,1H,J＝5.6Hz)，6.82(d,1H,J＝8.0Hz)，6.36(d,1H,J＝1.6Hz)，5.74(d,1H,J＝2.8Hz)，3.73(s,3H)，3.64(t,2H,J＝8.4Hz)，3.09(t,2H,J＝8.4Hz)。

LCMS:m/z 293.2(M+H)⁺；RT＝0.958min(254nm)。

Example 9

Synthesis of 2- (2-chlorophenyl) -1- (5- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) indolin-1-yl) ethanone

¹HNMR(400MHz,CDCl₃-d)8.41(d,1H,J＝5.2Hz)，8.31(d,1H,J＝8.8Hz)，7.91(s,1H)，7.86(d,1H,J＝8.4Hz)，7.49(d,1H,J＝1.6Hz)，7.43(d,1H,J＝2.0Hz)，7.42-7.24(m,3H)，7.17(d,1H,J＝5.2Hz)，6.80(s,1H)，6.35(d,1H,J＝2.0Hz)，4.23(t,2H,J＝8.8Hz)，3.95(s,2H)，3.81(s,3H)，3.30(t,2H,J＝8.4Hz)。

LCMS:m/z 445.4(M+H)⁺；RT＝1.36min(254nm)。

Example 10

Synthesis of 2- (3-chlorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) ethanone

¹H-NMR(CDCl₃,400MHz)：8..30-8.35(m,2H),7.88-7.91(m,2H),7.51-7.52(d,J＝1.2Hz,1H),7.29-7.32(m,3H),7.20-7.25(m,2H),6.41(d,J＝1.2Hz,1H),4.15-4.19(m,2H),3.85(s,3H),3.82(s,2H),3.25-3.29(m,2H).

LCMS:m/z444.9(M+H)；RT＝1.475min(2.50min).

Example 11

Synthesis of 2- (4-chlorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) ethanone

LCMS:m/z445.4(M+H)；RT＝1.39min(2.0min).

¹H-NMR(CDCl₃,400MHz):8.28-8.34(m,2H),7.88-7.91(m,2H),7.52(s,1H),7.33-7.35(m,2H),7.24-7.26(m,4H),6.42(s,1H),4.17(t,j＝8.0,2H),3.85(s,3H),3.81(s,2H),3.26(t,j＝8.0,2H).

Example 12

Synthesis of 3-hydroxy-1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) -2-phenylpropan-1-one

LCMS:m/z441.3(M+H)；RT＝1.15min(2.0min).

¹H-NMR(CDCl₃,400MHz):7.95-7.97(m,1H),7.83-7.85(m,1H),7.69(s,1H),7.52(s,1H)，7.31-7.39(m,5H),7.09-7.10(m,1H),6.43-6.54(m,2H),4.00-4.11(m,2H),3.85(s,3H),3.47-3.67(m,2H),3.43-3.61(m,1H),3.02-3.07(m,2H).

Example 13

Synthesis of 1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) -2-phenylpropan-1-one

¹H-NMR(CDCl₃,400MHz)：8..43-8.45(d,J＝7.6Hz,1H),7.19-8.20(d,J＝7.6Hz,1H),7.91-7.93(d,J＝8.4Hz,1H),7.81(s,1H),7.54-7.55(d,J＝2.0Hz,1H),7.27-7.37(m,6H),6.45-6.46(d,J＝2.0Hz,1H),4.17-4.22(m,1H),3.86-3.91(m,5H),3.04-3.12(m,2H),1.54-1.55(d,J＝6.8Hz,3H).

LCMS:m/z425.3(M+H)；RT＝1.462min(2.50min).

Example 14

2- (2-chlorophenyl) -N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) acetamide

¹H-NMR(CDCl₃,400MHz)：8.92(d,J＝2.0Hz,1H),7.99-8.01(d,J＝9.2Hz,1H),8.49-8.52(m,1H),8.44-8.46(d,J＝6.0Hz,1H),7.61-7.62(d,J＝3.6Hz,1H),7.7.43-7.46(dd,J₁＝3.6Hz,J₂＝7.2Hz,1H),7.37-7.39(dd,J₁＝3.6Hz,J₂＝7.2Hz,1H),7.28-7.32(m,3H),6.50-6.51(d,J＝2.4Hz,1H),4.01(s,2H),3.92(s,3H).

LCMS:m/z419.9(M+H)；RT＝1.375min(2.50min).

Example 15

Synthesis of 3N- (5- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) pyridin-2-yl) acetamide

¹HNMR(400MHz,CDCl₃-d)10.77(s,1H)，9.49(s,1H)，9.04(s,1H)，8.52(d,1H,J＝4.8Hz)，8.44(d,1H,J＝8.4Hz)，8.21(d,1H,J＝8.8Hz)，7.47(d,1H,J＝4.8Hz)，7.36(s,1H)，6.28(s,1H)，3.70(s,3H)，2.13(s,3H)。

LCMS:m/z 310.0(M+H)⁺；RT＝0.771min。

Example 17

Synthesis of N- (1-methyl-1H-pyrazol-5-yl) -4- (6- (phenethylamino) pyridin-3-yl) pyrimidin-2-amine

Compound A3(20mg,0.05mmol) was added to a dry 50mL one-necked flask, dissolved in tetrahydrofuran (5mL), and lithium aluminum hydride (5mg,0.13mmol) was slowly added thereto, followed by reaction at room temperature overnight. After LCMS detection, water was added to the reaction mixture, which was extracted with ethyl acetate, the organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated and passed through a reverse phase preparative column to give compound a17(5mg, white solid) in yield: 27 percent.

¹H-NMR(CDCl₃,400MHz):8.74(s,1H),8.35(d,j＝5.6,1H),8.06-8.08(m,1H),7.47-7.48(m,1H),7.30-7.34(m,2H),7.22-7.24(m,2H),6.95-7.09(m,2H),6.34-6.43(m,2H),4.93(s,1H),3.80(s,3H),3.62-3.66(m,2H),2.93-2.97(m,2H).

LCMS:m/z372.2(M+H)；RT＝1.08min(2.0min).

Example 18

Synthesis of 4N- (2-chloro-4- (2- (1-methyl-1H-pyrazol-5-ylamino) pyrimidin-4-yl) phenyl) -2-phenylacetamide

¹HNMR(400MHz,CDCl₃-d)8.44(d,1H,J＝8.8Hz)，8.32(d,1H,J＝5.2Hz)，7.92(d,1H,J＝2.0Hz)，7.78(dd,1H,J＝2.0Hz,8.8Hz)，7.72(s,1H),7.39-7.27(m,6H),7.03(d,1H,J＝6.0Hz)，6.86(s,1H)，6.24(d,1H,J＝2.4Hz)，3.73(s,2H)，3.70(s,3H)。

LCMS:m/z 418.9(M+H)⁺；RT＝1.504min。

Example 19

Synthesis of N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) -1-phenylmethanesulfonamide

¹H-NMR(CDCl3 400MHz):8.47-8.57(m,2H),8.18-8.21(m,1H),7.52-7.53(m,1H),7.30-7.33(m,1H),7.25-7.26(m,1H),7.00-7.23(m,5H),6.36(s,1H),4.46(s,2H),3.83(m,3H).

LCMS:m/z421.9(M+H)；RT＝1.14min(2.5min).

Example 20

Synthesis of 2- (2, 6-dichlorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) indolin-1-yl) ethanone

¹H-NMR(CDCl₃,400MHz):8.41(d,j＝5.2,1H),8.26(d,j＝8.8,1H),7.94(s,1H),7.84(d,j＝8.4,1H)，7.49-7.50(m,1H),7.36-7.38(m,2H),7.16-7.22(m,2H),6.81(m,1H),6.35-6.36(m,1H),4.34(t,j＝8.4,2H),4.16(s,2H),3.81(s,3H),3.37(t,j＝8.4,2H),2.80(s,1H).

LCMS:m/z479.3(M+H)；RT＝1.43min(2.0min).

Example 21

Synthesis of 2- (2-chlorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) ethanone

¹H-NMR(CDCl3 400MHz):8.90(s,1H),8.44-8.45(m,1H),8.29(s,1H),7.26-7.49(m,7H),6.39(s,1H),4.65-4.66(m,2H),4.18-4.19(m,2H),3.77(s,3H),3.21-3.23(m,2H).

LCMS:m/z446.2(M+H)；RT＝1.30min(2min).

Example 22

Synthesis of 2- (2, 6-dichlorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) ethanone

¹H-NMR(CDCl3 400MHz):8.81(s,1H),8.45(s,1H),8.13(s,1H),7.51(s,1H),7.33-7.35(m,2H),7.17-7.19(m,2H),6.96(s,1H),6.35(s,1H),4.88(s,2H),4.23(t,j＝8.4,2H),3.82(s,3H),3.18(t,j＝8.4,2H).

LCMS:m/z480.2(M+H)；RT＝1.41min(2min).

Example 23

Synthesis of N-methyl N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) -2-phenylacetamide

Example 24

Synthesis of 3-methyl-N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) butanamide

Example 25

Synthesis of 2- (3-chlorophenyl) -N- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) acetamide

Example 26

Synthesis of 1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2-yl) -3-phenylurea

Example 27

Synthesis of N- (2-cyano-4- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) phenyl) -2-phenylacetamide

Example 28

Synthesis of 2- (2-fluorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) ethan-1-one

Example 29

Synthesis of 2- (3-fluorophenyl) -1- (5- (2- ((1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) -2, 3-dihydro-1H-pyrrolo [2,3-b ] pyridin-1-yl) ethan-1-one

Comparative example C14 Synthesis of (6-aminopyridin-3-yl) -N- (1-methyl-1H-pyrazol-5-yl) pyrimidin-2-amine

A round-bottomed flask 25mL was charged with A15(180mg,0.58mmol), sodium hydroxide (116mg, 2.91mmol), methanol (5mL) and water (2mL) in that order. The reaction was stirred at 80 ℃ for 6h, TLC checked for completion, the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (dichloromethane: methanol 5:1) to give compound C1(124mg, yield: 80%) as a yellow solid.

¹HNMR(400MHz,CDCl₃-d)8.75(d,1H,J＝1.6Hz)，8.38(d,1H,J＝5.2Hz)，8.10(dd,1H,J＝2.4Hz,8.8Hz)，7.49(d,1H,J＝2.0Hz)，7.10(d,1H,J＝5.6Hz)，6.83(s,1H)，6.56(d,1H,J＝8.8Hz)，6.34(d,1H,J＝1.6Hz)，4.81(s,2H)，3.80(s,3H)。

LCMS:m/z 268.2(M+H)⁺；RT＝0.648min。

Comparative example C2

Preparation of 4- (6-aminopyridin-3-yl) pyrimidin-2-amine:

in a dry 50mL three-necked flask were added in the order compound C2-1(200mg, 1.45mmol), C2-2(188mg, 1.45mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (110mg, 0.15mmol), potassium carbonate (300mg, 2.17mmol), 1, 4-dioxane (8mL) and water (2 mL). Heating to 100 ℃ under the protection of nitrogen, and reacting for 3 hours. After completion of the reaction, the reaction mixture was poured into 30mL of water, extracted with ethyl acetate (30 mL. times.2), and the organic phases were combined. The organic phase was washed with saturated brine (50mL × 1) in this order, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure and purified with Flash (dichloromethane: methanol ═ 30: 1 to 10:1) to give product C2(120mg, light yellow solid) in yield: 30 percent.

¹HNMR(400MHz,MeOD-d₄)8.76(d,1H,J＝1.6Hz)，8.59(dd,1H,J＝2.0Hz,9.2Hz)，8.31(d,1H,J＝6.4Hz)，7.36(d,1H,J＝6.4Hz)，7.09(d,1H,J＝9.2Hz)。

LCMS:m/z188.1(M+H)；RT＝0.29min(2min).

Test example 1 determination of ERK kinase Activity by Compounds of the present invention

Materials and instruments

ERK2enzyme(PV3595,Invitrogen)

Kinase Assay Kit-Ser/Thr 3Peptide(PV3176)

Synergy 2Microplate Reader(BioTec)

ProxiPlate-384Plus F,Black 384-shallow well Microplate(Cat.6008269,PerkinElmer)

The test method comprises the following steps:

Z′-LYTE^TMSer/Thr 3Peptide Substrate, Phospho-Peptide, 5X Kinase Buffer, ATP, Development Reagent A, Development Buffer, Stop Reagent all reagents were equilibrated to room temperature in preparation for loading.

Screening concentrations to test compounds for their effect on ERK kinase activity were 3-fold gradient dilutions starting at 1 μ M (0.2 μ M), 8 concentrations, each concentration being taken in duplicate wells, using 4% DMSO as co-solvent. After the reaction was completed, 5. mu.l of Development Reagent A diluted with Development buffer was added to all reaction wells, and after 1 hour of reaction at room temperature, 5. mu.l of Stop Reagent was added to all reaction wells to terminate the reaction, and a fluorescent signal (excitation wavelength 400nm, emission wavelength 460nm, 528nm) was detected using a Synergy 2Microplate Reader.

The inhibition rate of each well was calculated from the total active and background signal wells. The experiment was repeated in parallel twice. IC50 values can be calculated from the inhibition of the kinase by the test compound at a range of different concentrations.

TABLE 1 inhibition of various kinase activities by the compounds of the invention

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. A compound of formula I, stereoisomers, racemates, or pharmaceutically acceptable salts thereof:

wherein R is₅Selected from the group consisting of: H. substitutionOr unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, -OH, cyano, halogen, amino, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted C1-C8 alkylcarbonyl, substituted or unsubstituted C1-C8 alkoxycarbonyl, substituted or unsubstituted C1-C8 carboxyl, substituted or unsubstituted C1-C8 ester, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

m is 0, 1,2 or 3;

2. A compound, stereoisomer, racemate, or pharmaceutically acceptable salt thereof, according to claim 1, wherein said substitution is with one or more (e.g. 1 to 3) substituents selected from the group consisting of: halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkylhydroxy, -OH, C1-C3 alkoxy, C1-C3 alkylamino, 3-8 membered cyclic hydrocarbon, 3-8 membered heterocyclic group, amino and nitro.

3. The compound of claim 1, stereoisomers, racemates or pharmaceutically acceptable salts thereof, wherein said compound of formula I is represented by the following formula Ia:

R₃selected from the group consisting of: H. substituted or unsubstituted C1-C8 alkyl, -OH, cyano, halogen, C1-C8 alkylenehydroxy, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl, substituted or unsubstituted 3-8 membered aryl, and substituted or unsubstituted3-8 membered heteroaryl;

R₂、R₄、R₅is as defined in claim 1.

4. The compound of claim 1, stereoisomers, racemates or pharmaceutically acceptable salts thereof, wherein the compound of formula I is represented by the following formula Ib:

wherein,

X₁、X₂、X₃、X₅、X₆each independently selected from CR₅Or N;

p is 0, 1,2,3 or 4;

q is 1,2,3,4 or 5;

and p + q is less than or equal to 5;

R₂、R₄、R₅is as defined in claim 1.

5. A compound of claim 1, stereoisomers, racemates or pharmaceutically acceptable salts thereof, wherein in said compound of formula Ia or Ib R₂Selected from the group consisting of:

wherein each Ra is independently selected from: C1-C4 alkyl;

n is 0, 1,2 or 3;

p is 0, 1,2,3 or 4;

q is 1,2,3,4 or 5;

and p + q is less than or equal to 5;

6. A pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the compounds of claim 1, stereoisomers, racemates, or pharmaceutically acceptable salts thereof and a pharmaceutically acceptable excipient.

7. Use of a compound according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, for the preparation of a medicament for the prevention and treatment of diseases associated with ERK kinase and targeted inhibitors of ERK kinase.

8. A process for preparing a compound of claim 1, comprising the steps of:

wherein, X₁、X₂、X₃、X₄、X₅、X₆、R₁、R₂、R₃The definition of each group is as defined in claim 1;

9. A process for preparing a compound of claim 1, comprising the steps of:

10. A method of non-therapeutically inhibiting ERK kinase activity comprising the steps of: contacting a compound of claim 1, or a pharmaceutically acceptable salt thereof, with ERK kinase, thereby inhibiting ERK kinase.