CN106279119A

CN106279119A - The preparation of a kind of Azaindole kinase inhibitors and application thereof

Info

Publication number: CN106279119A
Application number: CN201510279568.6A
Authority: CN
Inventors: 刘磊; 程鹏; 耿美玉; 曹文杰; 江磊; 艾菁; 黄敏; 杨新颖; 查传涛; 曹建华; 陈春麟; 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: 2015-05-27
Filing date: 2015-05-27
Publication date: 2017-01-04
Anticipated expiration: 2035-05-27
Also published as: WO2016188214A1; CN106279119B

Abstract

The invention discloses preparation and the application thereof of a kind of Azaindole kinase inhibitors.Specifically, the invention provides a kind of compound as shown in following formula I；Wherein, the definition of each group is as noted in the discussion.The compound of the present invention has good FGFR kinase inhibiting activity, may be used for preparing the medicine of the relevant disease of a series for the treatment of FGFR kinase activity.

Description

Preparation and application of novel kinase inhibitor

Technical Field

The invention belongs to the field of pharmaceutical chemistry; in particular, the invention relates to preparation and application of a novel kinase inhibitor.

Background

The receptor tyrosine kinase plays a key role in various links such as tumorigenesis development, invasion and metastasis, drug resistance and the like due to abnormal expression activation or gene mutation, and becomes an important target for research and development of antitumor drugs. Wherein, Fibroblast Growth Factor Receptors (FGFRs) are important members of receptor tyrosine kinase family, and mainly comprise four subtypes of FGFR1, FGFR2, FGFR3 and FGFR 4. The ligand is Fibroblast Growth Factors (FGFs). Due to gene amplification, mutation, fusion or ligand induction and the like, all the FGFR members are continuously activated to induce the proliferation, invasion and migration of tumor cells, promote angiogenesis and promote the generation and development of tumors. FGFRs are highly expressed and abnormally activated in various tumors and are closely related to poor prognosis of tumor patients, such as non-small cell lung cancer, breast cancer, gastric cancer, bladder cancer, endometrial cancer, prostate cancer, cervical cancer, colon cancer, esophageal cancer, keratinocyte tumor, myeloma, rhabdomyosarcoma and the like.

Research shows that the amplification of FGFR1 accounts for 20% of non-small cell lung cancer squamous cell carcinoma, and research on in-vitro proliferation, signal path and the like of lung cancer cell strains amplified by FGFR1 shows that the FGFR selective inhibitor can effectively inhibit the activation of FGFR1 signal path and the proliferation of cells. Among breast cancers, the amplification of the chromosome (8p 11-12) region where FGFR1 is located accounts for about 10% of ER positive patients, and the abnormal activation of FGFR2 signaling pathway caused by the amplification or mutation of FGFR2 gene related to the high expression of FGFR1mRNA and the poor prognosis of patients is mainly related to gastric cancer, triple negative breast cancer, endometrial cancer and the like. The amplification rate of FGFR2 in the gastric cancer tissues is 5% -10%. The 313 gastric cancer tissues are analyzed to show that the amplification of FGFR2 is obviously related to the tumor size, the local infiltration degree, the lymph node metastasis condition and the occurrence of distant metastasis, and the gastric cancer with the FGFR2 amplification is generally a progressive tumor, has poorer prognosis and relatively lower overall survival rate of patients. FGFR2 amplification accounted for 4% of refractory triple negative breast cancers. Endometrial cancer is a common gynecological genital tract tumor, and mutations in FGFR2 account for approximately 12% of endometrial cancer. 50% -60% of FGFR3 mutation in non-invasive bladder cancer and 10% -15% of FGFR3 mutation in invasive bladder cancer. FGFR3t (4; 14) (p 16.3; q32) gene rearrangements account for 15-20% in multiple myeloma. In addition, several preclinical and clinical studies have shown the importance of aberrant activation of the FGF/FGFR axis in liver cancer. FGFR of various subtypes and ligands thereof, FGFs, in liver cancer have abnormal expression and activation, such as FGFR2, FGFR3, FGFR4, FGF19, FGF2, FGF5, FGF8, FGF9 and the like.

It is not to be ignored that abnormal activation of the FGF/FGFR axis is closely related to resistance to EGFR inhibitors, neovascular inhibitors, endocrine therapy, and the like.

The development of targeted FGFR kinase inhibitors has become a leading hotspot in research of antitumor drugs. Therefore, there is a great need in the art to develop new FGFR kinase inhibitors.

Disclosure of Invention

The invention aims to provide a novel kinase inhibitor, and preparation and application thereof.

In a first aspect, the present invention provides a compound represented by formula I below, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof:

in the formula,

p is selected from: H. tetrahydropyranyl (THP), t-butyloxycarbonyl (Boc), 2- (trimethylsilyl) ethoxymethyl (SEM) or 4-methoxybenzyl (PMB);

x is selected from: H. cl, F, -CN or Me;

u, V, W, Z are each independently selected from: n or-CH-;

y is selected from: H. f, Cl or-OMe;

l is selected from:

(a)or

(b) -NH-, substituted or unsubstituted 6-8 membered aryl or heteroaryl, substituted or unsubstituted C1-C6 straight or branched chain hydrocarbyl, substituted or unsubstituted 5-8 membered saturated heterocyclyl or carbocyclyl, carbonyl, oximino, sulfonamido, a heteroatom selected from N, O or S;

q is selected from: -unsubstituted, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenylene or alkynylene, -NH, -O-, -S-;

ring A is selected from: a non-substituted or unsubstituted 6-8 membered arylene or heteroarylene group, a substituted or unsubstituted 3-12 membered saturated or unsaturated heterocyclyl or carbocyclyl group;

r is selected from: H. substituted or unsubstituted groups selected from the group consisting of:

wherein,

m is selected from: none, N, O, substituted or unsubstituted C1-C6 alkylene, substituted or unsubstituted C6-C10 arylene, or heteroarylene;

j is selected from: no, -OH, halogen, -NH₂Substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted 4-7 membered heterocyclic group, substituted or unsubstituted C1-C6 alkoxy, 5-12 membered bridged ring groupOr heterobridged ring groups; wherein the above groups are optionally substituted with one or more substituents selected from the group consisting of: halogen, -NH₂OH, -OH, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocyclyl, C1-C6 alkylthio; or the above groups and C3-C6 cycloalkyl form a spiro structure;

any of the above heterocyclyl, heteroaryl, heteroarylene, or heterobridged ring groups contains at least one heteroatom selected from the group consisting of: n, O or S;

any "substitution" described above means that one or more hydrogen atoms on the group is substituted with a substituent selected from the group consisting of: halogen, -OH, -NH₂CN, -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered saturated heterocycle or carbocycle; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O or S, the heterocycle comprising 1 to 3 heteroatoms selected from the group consisting of: n, O or S.

In another preferred embodiment, P, X, U, V, W, Z, Y, L, Q, ring A, R, M, J is the corresponding group in the particular compound described in the examples.

In another preferred embodiment, the ring a is a group selected from the group consisting of:

wherein Q is₁、Q₂、Q₃、Q₄Each independently selected from: n or-CH-;

B₁、B₂、B₃、B₄each independently selected from: n or-CH-.

in another preferred embodiment, R is a substituted or unsubstituted group selected from the group consisting of:

wherein,

R₁、R₂、R₃、R₄each independently selected from: H. halogen, C1-C6 alkyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five-or six-membered aryl optionally containing at least one heteroatom selected from N, O or S, or R1 and R2, R3 and R4 are each linked to form a 3-6 membered ring;

R₅selected from: H. -OH, C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S;

R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂each independently selected from: H. C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S, or R₆And R₇，R₈And R₉Respectively form 4-6 membered rings with carbon atoms;

G₁、G₂、G₃、G₄each independently selected from: H. halogen, C1-C6 alkyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five-or six-membered aryl optionally containing at least one heteroatom selected from N, O or S, or G1 and G2, and G3 and G4 are each linked to form a 3-6 membered ring;

G₅selected from: H. c1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S;

E₁、E₂each independently selected from: H. halogen, C1-C6 alkyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S;

E₃selected from: H. -OH, C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S;

R₁₃、R₁₄、R₁₅、R₁₆、R₁₇each independently selected from: H. C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S, or R₁₃And R₁₄，R₁₅And R₁₆Respectively form 4-6 membered rings with carbon atoms;

each of the foregoing groups may be optionally substituted with one or more substituents selected from the group consisting of: halogen, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 alkylthio, -NH₂-OH, trifluoromethyl and optionally a five or six membered aryl group comprising at least one heteroatom selected from N, O or S;

C0-C3 alkyl is alkylene having no, or 1-3, carbon atoms;

C1-C6 alkyl is alkylene having 1 to 6 carbon atoms.

In another preferred embodiment, P is selected from: H. tetrahydropyranyl (THP), t-butyloxycarbonyl (Boc), 2- (trimethylsilyl) ethoxymethyl (SEM) or 4-methoxybenzyl (PMB);

x is selected from: H. cl, F, -CN;

w, Y, Z are each independently selected from: n or-CH-;

ring A is selected from: a substituted or unsubstituted six membered aryl or heteroaryl group, wherein said heteroaryl group comprises at least one heteroatom selected from the group consisting of: n, O, S, respectively;

m is selected from the group consisting of: and (C) a non-substituted or unsubstituted C1-C4 alkylene group.

In another preferred embodiment, P is H;

x is selected from: H. cl, F;

w, Y, Z are each independently selected from: n or-CH-;

ring A is selected from: none, phenyl, pyrazolyl, pyridyl, thiazolyl, piperidinyl;

m is selected from: a non-substituted or unsubstituted C1-C3 alkylene group;

any of the above heterocycles, heteroaryls or heteroarylenes comprising at least one heteroatom selected from the group consisting of: n, O or S;

In another preferred embodiment, L is a group selected from the group consisting of:

wherein, B₁、B₂、B₃、B₄Is independently selected from: n or-CH-.

In another preferred embodiment, when Q is nothing, L is

In another preferred embodiment, L is not null when both Y are-OMe.

In another preferred embodiment, when both Y are-OMe, L is not

In another preferred embodiment, both Y's are not simultaneously-OMe.

In another preferred embodiment, neither Y is-OMe.

In another preferred embodiment, ring a is a group selected from the group consisting of: none, phenyl, pyrazolyl, pyridyl, thiazolyl, or piperidinyl.

In another preferred embodiment, M is selected from the group consisting of: and (C) a non-substituted or unsubstituted alkylene group of C1 to C3.

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

1) q is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -NH, -O-, -S-, and the compound of formula I is selected from the following group:

2)

2.1) when Q is none, L isWhen said compound of formula I is selected from the group consisting of:

2.2) when Q is none, L isWhen said compound of formula I is selected from the group consisting of:

2.3) when Q is absent, L is-NH-, carbonyl, substituted or unsubstituted 6-8 membered aryl or heteroaryl, substituted or unsubstituted 5-8 membered saturated heterocyclyl or carbocyclyl, substituted or unsubstituted C1-C6 straight or branched chain hydrocarbyl, sulfonamide or oximo, the compound of formula I is selected from the group consisting of:

in another preferred embodiment, any of the above heterocycles, heteroaryls or heteroarylenes comprises at least one heteroatom selected from the group consisting of: n, O or S;

in another preferred embodiment, any "substitution" described above means that one or more hydrogen atoms on the group is substituted with a substituent selected from the group consisting of: halogen, -OH, -NH₂CN, -unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated CA substituted or halogenated C2-C6 acyl group, an unsubstituted or halogenated 5-to 8-membered aryl group, an unsubstituted or halogenated 5-to 8-membered heteroaryl group, an unsubstituted or halogenated 4-to 8-membered saturated heterocyclic group or carbocyclic group; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O or S, the heterocycle comprising 1 to 3 heteroatoms selected from the group consisting of: n, O or S.

A second aspect of the invention provides a process for the preparation of a compound of formula I as described in the first aspect of the invention, said process comprising step (a):

reacting a compound of formula I-1 with a compound of formula I-2 in an inert solvent to form a compound of formula I-3;

the definition of each group in each formula is as described above.

In another preferred embodiment, in the step (a), the reaction is carried out in the presence of a condensing agent.

In another preferred embodiment, the condensing agent is selected from the group consisting of: HATU, HBTU, EDCI, CDI, DCC, DMC, or a combination thereof.

In another preferred embodiment, in the step (a), the reaction is carried out in the presence of a base.

In another preferred embodiment, the base is an organic base.

In another preferred embodiment, the base is selected from the group consisting of: triethylamine, DIPEA, or a combination thereof.

In another preferred embodiment, the inert solvent is selected from the group consisting of: toluene, dioxane, acetonitrile, DCM, THF, DMF, DMA, or combinations thereof.

In another preferred example, the method further comprises step (b):

carrying out deprotection reaction by using a compound shown as a formula I-3 in an inert solvent to obtain a compound shown as a formula I-4;

the definition of each group in each formula is as described above.

In another preferred embodiment, in the step (b), the reaction is carried out in the presence of an acid.

In another preferred embodiment, the acid is selected from the group consisting of: hydrochloric acid, p-toluenesulfonic acid, TFA, or combinations thereof.

In another preferred embodiment, in the step (b), the inert solvent is selected from the group consisting of: dichloromethane, methanol, ethanol, isopropanol, n-butanol, t-butanol, isobutanol, or combinations thereof.

A third aspect of the invention provides the use of a compound according to the first aspect of the invention for:

(a) preparing a medicament for treating diseases related to the activity or expression level of FGFR kinase;

(b) preparing an FGFR kinase targeted inhibitor;

(c) non-therapeutically inhibiting the activity of FGFR kinase in vitro;

(d) non-therapeutically inhibiting tumor cell proliferation in vitro; and/or

(e) Treating diseases related to the activity or expression level of FGFR kinase.

In another preferred embodiment, the FGFR kinase is selected from the group consisting of: FGFR1, FGFR2, FGFR3, or a combination thereof.

In a fourth aspect of the present invention, there is provided a pharmaceutical composition comprising:

(i) an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof; and

(ii) a pharmaceutically acceptable carrier.

In a fifth aspect, the present invention provides a method for inhibiting FGFR kinase activity, comprising the steps of:

administering to a subject in need thereof an inhibitory effective amount of a compound of formula I as described in the first aspect of the invention or a pharmaceutically acceptable salt thereof; or

Administering to the subject an inhibitory effective amount of a pharmaceutical composition according to the fourth aspect of the invention.

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 prepares a compound with a structure shown in a formula I through long-term and intensive research, and finds that the compound has FGFR kinase inhibitory activity. The compound has an inhibiting effect on a series of FGFR kinases at an extremely low concentration (which can be as low as less than or equal to 100nmol/L), and has extremely excellent inhibiting activity, so that the compound can be used for treating diseases related to the activity or expression level of the FGFR kinases, such as tumors. The present invention has been completed based on this finding.

Term(s) for

As used herein, the term "C1-C6 alkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

As used herein, the term "C2-C6 acyl" refers to a substituent such as a "straight or branched alkyl-carbonyl group having 1 to 6 carbon atoms" structure, such as acetyl, propionyl, butyryl, or the like.

The term "C1-C6 alkylene" refers to a group formed after a C1-C6 alkyl group has lost one hydrogen atom, as described above, for example-CH₂-、-CH₂-CH₂-, or the like.

The term "C6-C10 arylene" refers to an aryl group having 6 to 10 carbon atoms with one hydrogen atom removed to form a group, and includes monocyclic or bicyclic arylene groups such as phenylene, naphthylene, or the like.

The term "six-membered aryl" refers to phenyl.

The term "6-to 8-membered aryl" refers to a 6-to 8-membered ring substituent of a carbon unsaturated series, such as phenyl, or the like.

The term "six-membered to 8-membered heteroaryl" refers to a substituent of an unsaturated ring system having 6 to 8 members with one or more heteroatoms selected from O, S, N or P in the ring system, such as pyridyl, thienyl, or the like.

The term "saturated 3-to 12-membered carbocyclic ring" refers to a saturated carbocyclic ring having 3 to 12 carbon atoms, such as cyclohexyl, or the like.

The term "3-12 membered heterocyclic ring" refers to a saturated ring system substituent having one or more heteroatoms selected from O, S, N or P in a 3-12 membered ring system, such as piperidinyl, pyrrolyl, or the like.

The term "halogen" refers to F, Cl, Br and I.

As used herein, the terms "comprising," "including," or "including" mean that the various ingredients may be used together in a mixture or composition of the invention. Thus, the terms "consisting essentially of and" consisting of are encompassed by the term "comprising.

In the present invention, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response), i.e., at a reasonable benefit/risk ratio.

In the present invention, the term "effective amount" refers to an amount of a therapeutic agent that treats, alleviates, or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or prophylactic effect. The precise effective amount for a subject will depend upon the size and health of the subject, the nature and extent of the disorder, and the therapeutic agent and/or combination of therapeutic agents selected for administration. Therefore, it is not useful to specify an exact effective amount in advance. However, for a given condition, the effective amount can be determined by routine experimentation and can be determined by a clinician.

Herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on a group are replaced with a substituent selected from the group consisting of: halogen, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated C1-C6 alkyl-OH.

Unless otherwise specified, all occurrences of a compound in the present invention are intended to include all possible optical isomers, such as a single chiral compound, or a mixture of various chiral compounds (i.e., a racemate). In all compounds of the present invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

As used herein, the term "compounds of the invention" refers to compounds of formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula I.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic and organic salts. One preferred class of salts is that formed by reacting a compound of the present invention with an acid. Suitable acids for forming the salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

A compound of formula I

The invention provides a compound shown as the following formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, a diastereoisomer, a tautomer, a solvate, a polymorph or a prodrug thereof:

in the formula,

x is selected from: H. cl, F, -CN or Me;

u, V, W, Z are each independently selected from: n or-CH-;

y is selected from: H. f, Cl or-OMe;

l is selected from:

(a)or

wherein,

j is selected from: no, -OH, halogen, -NH₂Substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted 4-7 membered heterocyclyl, substituted or unsubstituted C1-C6 alkoxy, 5-12 membered bridged ring or heterobridged ring; wherein the above groups are optionally substituted with one or more substituents selected from the group consisting of: halogen, -NH₂OH, -OH, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocyclyl, C1-C6 alkylthio; or the above groups and C3-C6 cycloalkyl form a spiro structure;

any "substitution" described above means that one or more hydrogen atoms on the group is substituted with a substituent selected from the group consisting of: halogen, -OH, -NH₂CN, -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered saturated heterocycle or carbocycle; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O or S, the heterocyclic ring contains 1-3 heterocyclic rings selected from the group consisting ofThe hetero atom of (a): n, O or S.

wherein Q is₁、Q₂、Q₃、Q₄Each independently selected from: n or-CH-;

B₁、B₂、B₃、B₄each independently selected from: n or-CH-.

wherein,

R₅selected from: H. -OH, C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl or optionallyA five-or six-membered aryl group comprising at least one heteroatom selected from N, O or S;

G₅selected from: H. C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl, or a five or six membered aryl optionally containing at least one heteroatom selected from N, O or S;

each of the above groups may optionally be substitutedSubstituted with one or more substituents selected from the group consisting of: halogen, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 alkylthio, -NH₂-OH, trifluoromethyl and optionally a five or six membered aryl group comprising at least one heteroatom selected from N, O or S;

C0-C3 alkyl is alkylene having no, or 1-3, carbon atoms;

C1-C6 alkyl is alkylene having 1 to 6 carbon atoms.

x is selected from: H. cl, F, -CN;

w, Y, Z are each independently selected from: n or-CH-;

In another preferred embodiment, P is H;

x is selected from: H. cl, F;

w, Y, Z are each independently selected from: n or-CH-;

m is selected from: a non-substituted or unsubstituted C1-C3 alkylene group;

any "substitution" described above means that one or more hydrogen atoms on the group is substituted with a substituent selected from the group consisting of: halogen, -OH, -NH₂CN, -unsubstituted or halogenSubstituted C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered saturated heterocycle or carbocycle; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O or S, the heterocycle comprising 1 to 3 heteroatoms selected from the group consisting of: n, O or S.

wherein, B₁、B₂、B₃、B₄Are each independently selected from: n or-CH-.

In another preferred embodiment, when Q is nothing, L is

In another preferred embodiment, L is not null when both Y are-OMe.

In another preferred embodiment, when both Y are-OMe, L is not

In another preferred embodiment, both Y's are not simultaneously-OMe.

In another preferred embodiment, neither Y is-OMe.

2)

in another preferred embodiment, any "substitution" described above means that one or more hydrogen atoms on the group is substituted with a substituent selected from the group consisting of: halogen, -OH, -NH₂CN, -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered saturated heterocyclyl or carbocyclyl; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O or S, the heterocycle comprising 1 to 3 heteroatoms selected from the group consisting of: n, O or S.

Preparation of Compounds of formula I

The present invention provides a process for the preparation of a compound of formula I, said process comprising step (a):

the definition of each group in each formula is as described above.

In another preferred embodiment, the base is an organic base.

In another preferred example, the method further comprises step (b):

the definition of each group in each formula is as described above.

Use of compounds of formula I

The present invention provides the use of a compound of formula I:

(a) preparing a medicament for treating diseases related to the activity or expression quantity of the F GFR kinase;

(b) preparing an FGFR kinase targeted inhibitor;

(c) non-therapeutically inhibiting the activity of FGFR kinase in vitro;

(d) non-therapeutically inhibiting tumor cell proliferation in vitro; and/or

Pharmaceutical compositions and methods of administration

The compound has excellent inhibitory activity on FGFR Kinase (Kinase), such as FGFR1, FGFR2 and FGFR3, so the compound and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates and a pharmaceutical composition containing the compound as a main active ingredient can be used for treating, preventing and relieving diseases related to the activity or expression quantity of the FGFR. According to the prior art, the compounds of the invention are useful for the treatment of the following diseases: endometrial cancer, breast cancer, gastric cancer, bladder cancer, myeloma, liver cancer, and the like.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof in a safe and effective amount range and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of a compound of the invention per dose, more preferably, 5-200mg of a compound of the invention per dose. Preferably, said "dose" is a capsule or tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition can be blended with the compounds of the present invention and with each other without significant degradationThe efficacy of the compound. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g. tween, etc.)) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 5 to 500 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The main advantages of the invention include:

1. provides a compound shown as a formula I.

2. The FGFR inhibitor has a novel structure, and preparation and application thereof, and can inhibit the activity of various FGFR kinases at an extremely low concentration.

3. Pharmaceutical compositions for treating diseases associated with FGFR 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 by weight.

In each example:

LCMS apparatus: pump Agilent 1100 UV detector: agilent 1100 DAD

Mass Spectrometer API 3000

A chromatographic column: waters sunfire C18, 4.6X 50mm,5um

Mobile phase: a-acetonitrile B-H₂O(0.1％FA)

EXAMPLE 1 Synthesis of Compound 2

A100 mL autoclave was charged with 1(483mg, 0.91mmol), Pd (dppf) Cl₂(5mg, 10%), DMF (10.0mL) and methanol (10.0mL), followed by addition of triethylamine (137mg, 1.36mmol), and heating to 80 ℃ under a carbon monoxide gas pressure of 1.5MPa, overnight reaction. And (4) directly carrying out spin-drying reaction liquid column chromatography purification (a developing agent EA: PE ═ 1: 5). White solid 2(415mg, 98.6%) was obtained. LC MS 381(M + H)⁺，RT＝1.41min。

Example 2 synthesis of compound 3:

a dry 50mL round-bottom flask was charged with Compound 2(2.64g, 5.69mmol), THF (10.0mL) and water (10.0mL), then aqueous lithium hydroxide solution (0.96g/7.6mL,22.7mmol) was added dropwise at room temperature. Stir at room temperature overnight. TLC and LC-MS showed the reaction was complete. The reaction mixture was extracted with MTBE, the aqueous phase was adjusted to PH 4-3 with 6N HCl, extracted with DCM, and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was spin dried to give the crude product, which was slurried with EA: PE ═ 1:1 to give compound 3(2.03, 79%) as a yellow solid. LCMS:453(M + H)⁺，RT＝1.804min.

EXAMPLE 3 Synthesis of Compound 4

In a dry 25mL round bottom flask, compound 3(1.0g, 2.22mmol), DIEA (573mg, 4.44mmol), HATU (1.3g, 3.33mmol) and DMF (10.0mL) were added and stirred at room temperature for 30 min. Compound 3a (499mg, 2.44mmol) was added and stirred at room temperature overnight. TLC and LC-MS showed the reaction was complete. The reaction was poured into water, extracted with DCM, the organic phases combined, dried with saturated brine, dried over anhydrous sodium sulfate and purified by column chromatography (DCM: MeOH ═ 10:1) to give compound 4(1.226g, 86%) as a white solid. LCMS 638(M + H)⁺，RT＝1.44min.

EXAMPLE 4 Synthesis of Compound 5

A dry 25mL round bottom flask was charged with 4(424mg) and dichloromethane (5.0mL), followed by dropwise addition of MeOH/HCl (10.0mL) and stirring overnight at room temperature. TLC and LC-MS showed the reaction was complete. The reaction was spin dried and slurried with EA: PE ═ 1:5 to give a white solid. Then slurried with saturated aqueous sodium bicarbonate, filtered, washed with water, and dried to give 5 as a white solid (210mg, 57%). LCMS:554(M + H)⁺，RT＝1.302min。

Compound 5: 6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (4- ((3R, 5S) -3, 5-dimethylpiperazin-1-yl) phenyl) -1H-indazole-3-carboxamide

¹H NMR(CDCl₃,400MHz)ppm 10.44(s,1H),8.77(s,1H),8.54(d,2H,J＝8.0Hz),7.64(d,2H,J＝8.0Hz),7.39(s,1H),7.18(d,1H,J＝8.0Hz),6.96(d,2H,J＝8.0Hz),6.65(s,1H),3.98(s,6H),3.51(d,2H,J＝11.2Hz),3.13-3.03(m,2H),2.31(t,2H,J＝2.2Hz),1.16(d,6H,J＝6.4Hz).

Using a similar procedure the following compounds were obtained:

EXAMPLE 5 Synthesis of Compound 6

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (4- (4-ethylpiperazin-1-yl) phenyl) -1H-indazole-3-carboxamide

¹H NMR(d6-DMSO,400MHz)ppm 8.26(s,1H),7.76(s,2H),7.48(s,1H),4.02(s,4H),3.94(s,6H),2.75-3.35(m,8H),1.24(s,3H).

EXAMPLE 6 Synthesis of Compound 7

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (4- (4-methylpiperazin-1-yl) phenyl) -1H-indazole-3-carboxamide

¹H NMR(d6-DMSO,400MHz)ppm 13.86(s,1H),10.26(s,1H),9.74(brs,1H),8.27(d,1H,J＝8.4Hz),7.80(d,2H,J＝8.8Hz),7.48(s,1H),7.07(d,1H,J＝8.4Hz),7.01(m,3H),3.97(s,6H),3.81(d,2H,J＝12.4Hz),3.52(d,2H,J＝10.0Hz),3.09(s,2H),2.93(t,2H,J＝12.0Hz),2.87(s,3H).

EXAMPLE 7 Synthesis of Compound 9

A100 mL autoclave was charged with 8(300mg, 0.62mmol), Pd (dppf)₂Cl₂(30.87mg, 10%) and triethylamine (0.133mL, 1.81 mmol) followed by 7mL of DMF and 7mL of CH₃OH, introducing CO of 1.5atm, stirring at 70 deg.C for 12 hr, adding the reaction solution20mL of water was extracted with ethyl acetate, and the resulting extract was subjected to spin-dry column chromatography to give Compound 9(285mg, 99%). LCMS:466(M + H)⁺,RT＝1.626min。

EXAMPLE 8 Synthesis of Compound 10

A50 mL round-bottomed flask was charged with Compound 9(285mg, 0.62mmol), 20mL THF was added, after stirring to dissolve it, 3N aqueous LiOH (30mL) was added, the mixture was stirred at room temperature for 15 hours, 10mL diethyl ether was added after the reaction was completed, the ether phase was removed, the aqueous phase was adjusted to pH7 with 1N dilute hydrochloric acid, followed by extraction with diethyl ether three times, and the combined organic phases were dried by spin-drying to give Compound 10(170mg, 60%). LCMS 450(M-H)^-,RT＝1.644min.

EXAMPLE 9 Synthesis of Compound 11

In a dry 25mL three-necked flask, compound 10(60mg, 0.13mmol), HATU (75.81mg, 0.2mmol), DIEA (34.3mg, 0.27mmol) and DMF (5mL) were added and the mixture was stirred at room temperature for 0.5 h, then 4- ((3S,5R) -3, 5-dimethylpiperazine) -aniline (27.1mg, 0.13mmol) was added and stirred at room temperature for 2h, 20mL of water was added, extraction was performed with ethyl acetate, the organic phases were combined and spin dried, and column chromatography gave compound 11(82mg, 98%). LCMS 639(M + H)⁺,RT＝1.644min.

EXAMPLE 10 Synthesis of Compound 12

In a dry 25mL round bottom flask was added 11(82mg, 0.13mmol), 10mL dichloromethane was added and dissolved with stirring, 0.5mL TFA was slowly added dropwise at room temperature, the mixture was reacted at room temperature for 18 hours to prepare compound 12(39mg, 54%) LCMS:555(M + H) in reverse phase⁺,RT＝1.626min。

¹H NMR(d6-DMSO,400MHz)ppm 14.45(s,1H),10.39(s,1H),9.01(s,1H),8.65(t,1H,J＝14.4Hz),8.39(m,1H),7.81(d,2H,J＝8.4Hz),7.33(t,1H,J＝14.8Hz),7.05(s,6H),3.86(d,2H,J＝5.2Hz),3.98(s,6H),1.29(t,8H,J＝6.4Hz).

Using a similar procedure the following compounds were obtained:

EXAMPLE 11 Synthesis of Compound 13

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (4- (4-ethylpiperazin-1-yl) phenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(d6-DMSO,400MHz)ppm 10.30(s,1H),8.66(d,1H,J＝7.6Hz),7.73(d,2H,J＝8.4Hz),7.29(d,1H,J＝7.6Hz),7.06(s,1H),6.93(t,2H,J＝4.0Hz),3.99(s,6H),3.11(m,1H),2.37(m,2H),1.03(t,3H,J＝6.4Hz).

EXAMPLE 12 Synthesis of Compound 14

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (4- (4-methylpiperazin-1-yl) phenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹HNMR(d6-DMSO,400MHz)ppm 14.45(s,1H),10.39(s,1H),10.07(s,1H),8.66(d,1H,J＝8.0Hz),7.79(d,2H,J＝8.8Hz),7.32(d,1H,J＝5.6Hz),7.07(s,1H),7.04(d,2H,J＝9.2Hz),3.99(s,6H),3.82(d,2H,J＝12.8Hz),3.46(m,2H),2.94(m,2H),2.85(s,2H).

EXAMPLE 13 Synthesis of Compound 15

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (6- ((3S,5R) -3, 5-dimethylpiperazin-1-yl) pyridin-3-yl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(d6-DMSO,400MHz)ppm 14.50(s,1H),10.55(s,1H),9.07(d,1H,J＝10.4Hz),8.65(m,2H),8.48(d,1H,J＝11.2Hz),8.12(m,1H),7.32(d,1H,J＝8.4Hz),7.05(d,2H,J＝9.2Hz),4.41(d,2H,J＝13.2Hz),4.00(s,2H),3.46(m,6H),2.75(m,2H),1.29(m,6H).

EXAMPLE 14 Synthesis of Compound 16

N- (2-chloro-4- ((3S,5R) -3, 5-dimethylpiperazin-1-yl) phenyl) -6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(d6-DMSO,400MHz)ppm 14.55(s,1H),9.88(s,1H),9.23(d,1H,J＝2.8Hz),8.67(t,1H,J＝6.0Hz),8.54(t,1H,J＝10.0Hz),7.83(t,1H,J＝12.0Hz),7.36(d,1H,J＝6.4Hz),7.26(s,1H),7.09(s,2H),4.00(s,2H),4.02(m,9H),2.72(m,3H),1.26(m,6H).

EXAMPLE 15 Synthesis of Compound 17

6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -N- (3- (((3S, 5R) -3, 5-dimethylpiperazin-1-yl) methyl) phenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(d-CDCl₃,400MHz)ppm 13.11(br,1H),9.11(s,1H),8.84(d,J＝8.0Hz,1H),8.19-8.17(m,1H),7.55(s,1H),7.42-7.38(m,1H),7.31(d,J＝8.0Hz,1H),7.09(d,J＝8.0Hz,1H),6.73(s,1H),4.10(s,2H),3.99(s,6H),3.88-3.82(m,2H),3.46-3.42(m,2H),3.18-3.11(m,3H),1.29(d,J＝8.0Hz,6H).

EXAMPLE 16 Synthesis of Compound 19

LiAlH was added sequentially to a dry 25mL round bottom flask₄(41mg, 1.1mmol) and THF (5.0 mL). N is a radical of₂A solution of compound 18(415mg, 0.89mmol) in THF (2.0mL) was added dropwise to the round bottom flask under ice-water bath conditions with protection. Then the reaction was carried out for 2h under ice-water bath. TLC and LC-MS showed the reaction was complete. Sequentially adding H₂O(50μL)，15％NaOH(50μL)，H₂O (150. mu.L) was added to the reaction solution and stirred for 10min, respectively. Dried over anhydrous sodium sulfate. The solvent was dried by evaporation to give crude compound 19 as a foamy white solid (385mg, 98.7%). LC MS 437(M + H)⁺,RT＝1.524min.

EXAMPLE 17 Synthesis of Compound 20

In a dry 25mL round bottom flask was added compound 19(366mg, 0.84mmol) and DMSO (5.0mL), followed by IBX (323mg, 1.25mmol) and stirred at room temperature for 2.0 h. TLC and LC-MS showed the reaction was complete. The reaction solution was poured into 10.0mL of ice water, extracted with EA, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was dried by spinning and then slurried with petroleum ether to give compound 20(284mg, 78%) as a pale yellow solid. LCMS 435(M + H)⁺,RT＝1.934min.

EXAMPLE 18 Synthesis of Compound 21

A dry 25mL round bottom flask was charged with Compound 20(284mg, 0.65mmol) and dry THF (5.0mL), then 20a (0.8N, 2.45mL) in THF was added dropwise under ice-water bath conditions. Then stirred for 2.0h under ice-water bath conditions. TLC and LC-MS showed the reaction was complete. Saturated NH₄HCl (3.0mL) was added to the reaction and stirred for 10 min. Then, the mixture was extracted with ethyl acetate, washed with saturated brine, and the organic phases were combined and dried over anhydrous sodium sulfate. Column chromatography purification (EA: PE ═ 1:5) afforded compound 21(308mg, 88.6%) as a white solid. LCMS:533(M + H)⁺，RT＝1.60min。

EXAMPLE 19 Synthesis of Compound 22

In a dry 15mL round bottom flask was added compound 21(308mg, 0.58mmol) and dichloromethane (10.0mL), followed by MnO₂(303mg, 3.48mmol) and stirred at room temperature overnight. TLC and LC-MS showed reactionAnd (6) finishing. The reaction solution was filtered, spin-dried and purified by column chromatography directly (EA: PE ═ 10:1) to give compound 22(235mg, 77%) as a white solid. LCMS:529(M + H) +, RT ═ 1.81min.

EXAMPLE 20 Synthesis of Compound 23

A dry 25mL round bottom flask was charged with 22(233mg, 0.442mmol), 22a (147mg, 1.3mmol), K₂CO₃(152mg, 1.1mmol) and DMSO (5.0mL), the reaction was heated to 138 ℃ and stirred overnight. TLC and LC-MS showed the reaction was complete. The reaction was cooled to room temperature, poured into ice water, extracted with EA, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography (DCM: MeOH ═ 100:1) to give compound 23(177mg, 65%) as a yellow solid. LCMS 623(M + H)⁺，RT＝1.41min.

EXAMPLE 21 Synthesis of Compound 24

A dry 25mL round bottom flask was charged with 23(50mg, 0.08mmol) and pyridine (5.0mL), followed by hydroxylamine hydrochloride (11mg, 0.16mmol), and stirred at room temperature overnight. TLC and LC-MS showed the reaction was complete. The reaction solution was spin dried, slurried with water, and filtered. After drying, compound 24 was obtained as a white solid (35mg, 69%). LC MS 639(M + H)⁺，RT＝0.78min.

EXAMPLE 22 Synthesis of Compound 25

A dry 25mL round bottom flask was charged with 24(35mg) and dichloromethane (5.0mL), followed by dropwise addition of MeOH/HCl (5.0mL) and stirring at room temperature overnight. TLC and LC-MS showed the reaction was complete. The reaction solution was spin-dried and purified. Compound 25(13mg, 43%) was obtained as a white solid after lyophilization. LCMS:554(M + H)⁺RT＝1.42min.

Compound 25: (Z) - (6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -1H-indazol-3-yl) (4- ((3R, 5S) -3, 5-dimethylpiperazin-1-yl) phenyl) methanone oxime

¹H NMR(d6-DMSO,400MHz)ppm 11.35(d,1H,J＝9.2Hz),9.11(s,1H),8.53-8.39(m,1H),7.45-7.36(m,2H),7.06(d,1H,J＝0.4Hz),7.03-7.00(m,2H),6.93(d,2H,J＝8.4Hz),4.18(d,1H,J＝18.8Hz),3.92(s,1H),2.70(dd,2H,J＝25.2Hz),2.63(s,3H),2.33(s,1H),1.30-1.24(m,6H),0.87-0.81(m,1H).

EXAMPLE 23 Synthesis of Compound 26

A dry 50mL round bottom flask was charged with 23(30mg, 0.048mmol), TFA (17mg, 0.145mmol) and dichloromethane (5.0mL) and stirred at room temperature overnight. The reaction was spun dry and slurried with EA: PE ═ 1:8 to afford compound 26(14mg, 53%) as a yellow solid. LCMS:539(M + H)⁺,RT＝1.51min.

Compound 26: (6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -1H-indazol-3-yl) (4- ((3R, 5S) -3, 5-dimethylpiperazin-1-yl) phenyl) methanone

1HNMR(CDCl3,400MHz)ppm 10.53(s,1H),9.22(s,2H),8.46(d,1H,J＝8.4Hz),8.28(d,2H,J＝8.4Hz),7.49(s,1H),7.22(d,1H,J＝8.4Hz),6.93(d,2H,J＝8.4Hz),6.66(s,1H),3.99(s,6H),3.81(d,2H,J＝12.8Hz),3.40(s,2H),3.00(t,2H,J＝24Hz),1.39(d,6H,J＝6.4Hz),0.88～0.83(m,2H).

EXAMPLE 24 Synthesis of Compound 27

A dry 50mL round bottom flask was charged with 23(40mg, 0.064mmol) and anhydrous THF (5mL), and methyl magnesium bromide (3.0M, 43. mu.L, 0.128mmol) was slowly added dropwise and stirred at room temperature overnight. LC-MS showed 1 to be incomplete. And 2.0 equivalent of methyl magnesium bromide is added, and the mixture is stirred for 2 hours at room temperature. LC-MS showed the reaction was complete. The reaction was quenched with 5.0mL of a saturated aqueous solution of ammonium chloride, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was dried by spinning to give crude compound 27 which was used directly in the next step.

LCMS:639(M+H)⁺,RT＝1.62min。

EXAMPLE 25 Synthesis of Compounds 28, 29

Compound 27(30mg) and dichloromethane (5.0mL) were added sequentially to a dry 50mL round-bottomed flask, and TFA (1.0mL) was added slowly dropwise under ice-water bath conditions, and stirred at room temperature overnight. LC-MS and TLC showed the reaction was complete. The reaction was purified by spin-drying to give pale yellow solids 28 and 29 (4.0 mg each, 16%).

LCMS:539,537(M+H)⁺,RT＝1.51mim，1.53min.

Compound 28: 6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -3- (1- (4- ((3R, 5S) -3, 5-dimethylpiperazin-1-yl) phenyl) ethyl) -1H-indazole

¹H NMR(DMSO,400MHz)ppm 8.39(d,2H,J＝8.8Hz),8.35(d,1H,J＝8.4Hz),7.45(s,1H),7.16(d,1H,J＝9.2Hz),7.12(d,2H,J＝8.4Hz),6.90(s,1H),4.19(d,2H,J＝15.6Hz),3.98(s,6H),3.97(s,1H),3.57～3.46(m,3H),2.86(t,3H,J＝17.2Hz),2.21～2.14(m,1H),2.06～1.99(m,1H),1.42(d,6H,J＝6.4Hz),0.90(t,1H,J＝13.2Hz),0.096(s,1H).

Compound 29: 6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -3- (1- (4- ((3R, 5S) -3, 5-dimethylpiperazin-1-yl) phenyl) vinyl) -1H-indazole

¹H NMR(d6-DMSO,400MHz)ppm 7.39(t,3H,J＝17.6Hz),7.33(s,1H),7.05(d,2H,J＝8.8Hz),6.88(s,1H),6.83(d,1H,J＝8.4Hz),5.71(d,2H,J＝8.4Hz),3.98(s,1H),3.97(s,6H),3.93(s,1H),3.57～3.45(m,3H),2.72(t,2H,J＝6.4Hz),2.06～1.99(m,1H),1.39(d,6H,J＝6.4Hz),0.93～0.86(m,1H).

EXAMPLE 26 Synthesis of Compound 31

A dry 50mL round bottom flask was charged with 30(870.0mg, 2.33mmol) and dichloromethane (15.0mL), and TFA (3.0mL) was added slowly dropwise over an ice water bath and stirred at room temperature for 18.0 h. Spin-drying the solvent to obtain a crude product, diluting the crude product with water, and saturating NaHCO₃The solution was adjusted to pH 8, extracted with ethyl acetate, the organic phase washed with water, dried over anhydrous sodium sulfate, and the solvent dried to give crude product 31(953mg, 80%) as a yellow solid, which was used in the next step without purification.

LCMS:290.0(M+H)⁺,RT＝1.293min.

EXAMPLE 27 Synthesis of Compound 32

In a dry 100mL round bottom flask was added compound 31(895mg, 3.1mmol), aqueous NaOH (10.3mL, 3N) and 1, 4-dioxane (5.0 mL). A solution of iodine (944.0mg, 1.2mmol) in 1, 4-dioxane was added dropwise at zero degrees. Stir at room temperature for 18.0 h. The reaction was quenched with saturated sodium thiosulfate solution, adjusted to pH 8 with aqueous HCl (2N), filtered, and the filter cake was slurried with water to give compound 32 as a yellow solid (822.0mg, yield: 63.9%). LCMS 415.9(M + H) +, RT 1.667 min.

EXAMPLE 28 Synthesis of Compound 33

A dry 50mL round bottom flask was charged with 32(822.0mg, 1.98mmol), p-TSA (76.0mg, 0.40mmol) and dichloromethane (15.0mL), DHP (333.0mg, 3.96mmol) was slowly added dropwise, and stirred at room temperature for 18.0 h. Diluting the reaction solution with water, extracting with dichloromethane, mixing organic phases, drying with anhydrous sodium sulfate, spin-drying the solvent to obtain crude product, and performing column chromatography (ethyl acetate: petroleum ether ═ 1:10) to obtain yellow solid compound 33(340.0mg, yield: 34.0%) LC MS:501.5(M + H)⁺,RT＝2.133min。

EXAMPLE 29 Synthesis of Compound 34

A dry 25mL three-necked flask was charged with 33(200.0mg, 0.40mmol), 33a (112mg, 0.48mmol), 33b (114.0mg, 0.80mmol), K₃PO₄(254.0mg, 1.2mmol), CuI (76mg, 0.40mmol) and dry DMF (2.0mL) were stirred at 120 ℃ for 6.0 h. The reaction mixture was diluted with water (20.0mL), extracted with ethyl acetate, the organic phases were combined, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was dried by rotary evaporation to give a crude product, which was P-TLC to give compound 34(133.0mg, yield: 54.7%) as a yellow solid. LCMS:605.1(M + H)⁺,RT＝1.113min。

EXAMPLE 30 Synthesis of Compound 35

A dry 25mL round bottom flask was charged with 34(130.0mg, 0.21mmol) and dichloromethane (5.0mL), and TFA (1.0mL) was added slowly dropwise over an ice water bath and stirred at room temperature for 4.0 h. The solvent was dried by rotary evaporation to give the crude product, which was prepared by acidic prep-HPLC to give Compound 35 as a yellow solid (53.4mg, TFA salt, yield: 39.2%). LCMS:521.01(M + H)⁺,RT＝1.243min.

¹H NMR(d6-DMSO)ppm 12.94(brs,1H),10.61(s,1H),9.08(m,1H),8.67(d,2H,J＝8.8Hz),8.47(m,1H),8.03(d,2H,J＝8.8Hz),7.77(d,1H,J＝8.4Hz),7.70(s,1H),7.56(d,1H,J＝16.8Hz),7.49(d,1H,J＝8.8Hz),7.26(d,1H,J＝16.8Hz),7.14(d,1H,J＝9.2Hz),4.12(m,2H),2.76(m,2H),1.29(d,6H,J＝6.4Hz).

EXAMPLE 31 Synthesis of Compound 38

A dry 250mL round bottom flask was charged with 36(1.97g, 0.01mol), TFA (114mg, 1mmol) and dichloromethane (10.0mL), DHP (1.26g, 0.01mol) was slowly added dropwise, and stirred at room temperature for 4.0 h. The reaction was diluted with 100.0mL of water, extracted with dichloromethane, the organic phases combined, dried over anhydrous sodium sulfate and the solvent dried to give compound 38(3.0g, 92%) which was used in the next step without further purification. LCMS 281(M + H)⁺。

EXAMPLE 32 Synthesis of Compound 40

A dry 250mL round bottom flask was charged with Compound 38(3.0g, crude), 39(3.8g,0.015mol), KOAc (1.96g,0.02mmol), Pd (dppf) Cl₂(110mg,1.5mmol), dioxane (30mL) was replaced with nitrogen 3 times, and then the mixture was stirred at reflux overnight. After the reaction was complete, the dioxane was spun off, EA (30mL x3) and water (50mLx3) were added and extracted separately, the organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography with spin-dry solvent to give compound 40(2.18g, Y: 66%, two steps). LCMS:329(M + H)⁺。

EXAMPLE 33 Synthesis of Compound 43

A dry 250mL round bottom flask was charged with Compound 40(2.18g,6.6mmol),42(2.3g,9.9mmol), Cs₂CO₃(4.3g,13.2mmol),41Pd(PPh₃)₄(76mg,0.066mmol), toluene (30mL) was replaced with nitrogen gas 3 times, and then the mixture was stirred at reflux overnight. After completion of the reaction, EA (30 mL. times.3) and water (50 mL. times. 3) were added for extraction and separation, and the organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography using a dry solvent to obtain Compound 43(1.78g, Y: 76.7%). LCMS:353(M + H)⁺.

EXAMPLE 34 Synthesis of Compound 44

Compound 43(900mg, 2.55mmol) was dissolved in a 250mL three-necked flask, a dichloromethane solution of sulfonyl chloride was slowly added to solution 43 by dropping at-10 deg.C, the temperature was slowly raised to room temperature, after stirring for 3h.LCMS to monitor completion of the reaction, 50mL of water was added, the combined organic phases were extracted, dried over anhydrous sodium sulfate, and the solvent was dried by spinning to give compound 44(900mg, Y: 84%) which was used in the next step without purification. LC MS 421(M + H)⁺.

EXAMPLE 35 Synthesis of Compound 45

In a dry 250mL round bottom flask, compound 44(900mg, crude) and TFA (20.0mL) were added and stirred at 0 ℃ to room temperature for 16.0 h. The reaction solution was spin-dried to give 700mg of Compound 45(Y: 97.2%) which was used in the next step without purification. LCMS 337(M + H)⁺。

EXAMPLE 36 Synthesis of Compound 46

To a dry 250mL round bottom flask was added compound 45(700mg, 2.08mmol), I₂(580mg, 2.3mmol), NaOH (1.84g, 46.0mmol) and 1, 4-dioxane (20.0 mL). Stir at rt for 2.0 h. Dichloro for reaction liquidExtraction with methane, washing with saturated sodium thiosulfate solution, combining the organic phases, drying over anhydrous sodium sulfate, and spin-drying the solvent to give compound 46(900g, 93%). LCMS:463(M + H) +.

EXAMPLE 37 Synthesis of Compound 47

A dry 250mL round bottom flask was charged with 46(900mg, 1.9mmol), TFA (22mg, 0.19mmol) and dichloromethane (50.0mL), DHP (243g, 2.9mmol) was added slowly dropwise and stirred at room temperature for 4.0 h. The reaction mixture was diluted with 50.0mL of water, extracted with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was dried to give Compound 47(670mg, 63%). LCMS:547(M + H)⁺.

EXAMPLE 38 Synthesis of Compound 48

A dry 250mL round bottom flask was charged with Compound 47(100mg,0.183mmol),47a (64mg,0.274mmol), K₃PO₄(156mg,0.732mmol), CuI (36mg,0.189mmol),47b (52mg,0.732mmol), and DMF (5mL) were replaced with nitrogen for 3 times, and then stirred at reflux overnight. After completion of the reaction, EA (30mLx 3) and water (50mLx3) were added for extraction and separation, the organic phases were combined, dried over anhydrous sodium sulfate and the solvent was dried to give compound 48, which was used in the next step without further purification, 200mg crude, LCMS:652(M + H)⁺.

EXAMPLE 39 Synthesis of Compound 49

In a dry 250mL round bottom flask, compound 48(200mg, crude) and TFA (20.0mL) were added and stirred at 0 ℃ to room temperature for 16.0 h. The reaction solution was dried by evaporation to give 15mg of compound 49 (yield 14%). LCMS 568(M + H)⁺。

¹H NMR(d6-DMSO,400MHz)ppm12.46(br,1H),10.43(br,1H),8.25(m,1H),7.95(d,J＝8.4Hz,2H),7.58(d,J＝8.8Hz,1H),7.02(d,J＝8.8Hz,2H),6.96-6.92(m,2H),4.40(s,2H),3.95(s,6H),3.85(d,J＝8.0Hz,2H),2.99(m,2H),2.42-2.36(m,2H),1.97(br,1H),1.11(d,J＝6Hz,6H).

EXAMPLE 40 Synthesis of Compound 51

A dry 250mL round bottom flask was charged with Compound 50(100mg,0.188mmol),50a (38.8mg,0.188mmol), Pd₂(dba)₃(17.2mg,0.0188mmol),xantphos(11mg,0.0188mmol),Cs₂CO₃(123mg,0.376mmol), dioxane (20mL) was replaced with nitrogen 3 times, and then the mixture was stirred at reflux overnight. After completion of the reaction, EA (30mL x3) and water (50mLx3) were added for extraction separation, the organic phases were combined, dried over anhydrous sodium sulfate and the solvent was dried to give compound 51, which was used in the next step without further purification. This gave 150mg of Compound 51 (crude) LC MS 639(M + H)⁺.

EXAMPLE 41 Synthesis of Compound 52

In a dry 250mL round bottom flask, compound 51(150mg, crude) and TFA (20.0mL) were added and stirred at 0 ℃ to room temperature for 16.0 h. The reaction solution was dried by evaporation to give 46mg of compound 52 (46%).

LCMS:528(M+H)⁺,¹H NMR(d6-DMSO,400MHz)ppm12.15(br,1H),9.15-9.13(m,1H),8.99(s,1H),8.95(s,2H),8.58-8.55(m,1H),8.02(d,J＝8.0Hz,1H),7.22(s,1H),7.06(s,1H),6.89(d,J＝8.0Hz,1H),4.75(d,J＝12Hz,2H),4.03(s,6H),2.90-2.83(t,J＝12Hz,2H),1.35(d,J＝4Hz).

EXAMPLE 42 Synthesis of Compound 54

53(890mg,3.8mmol), hexabutylditin (2.65g, 4.6mmol), lithium chloride (958mg,22.8mmol), Pd2dba3(174mg,0.19mmol), tricyclohexylphosphine (128mg,0.46mmol) and 6mL dioxane were added to a microwave reaction flask. The air in the flask was replaced with nitrogen three times, and then microwave-reacted at 140 ℃ for 2 hours. The solvent was spun dry and the crude product was purified to give compound 54(750mg, 35%). LC MS, RT 1.532mim.

EXAMPLE 43 Synthesis of Compound 55

Compound 54(750mg,1.5mmol), 1,2,4, 5-tetrafluoro-3-bromobenzene (341mg,1.5mmol), Pd (PPh) were added to a microwave reaction flask₃)₄(173mg, 0.15mmol) and toluene (15.0 mL). The air in the flask was replaced with nitrogen three times, and then microwave-reacted at 130 ℃ for 2 hours. The solvent was spun dry and the crude product was purified to give compound 55(310mg, 59%). LC MS:352(M + H) +, RT ═ 1.518mim.

EXAMPLE 44 Synthesis of Compound 56

In a dry 100mL round bottom flask was added compound 55(310mg, 0.88mmol),4M HCl/MeOH (6mL) and methanol (6 mL). Stir at room temperature overnight. The organic solvent was spin dried and the crude product was taken up with saturated NaHCO₃Neutralized and then extracted twice with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate and spun to give compound 56(185mg, 79%). LC MS:268(M + H) +, RT ═ 0.998mim.

EXAMPLE 45 Synthesis of Compound 57

In a dry 100mL round bottom flask were added compound 56(185mg, 0.69mmol), NIS (187mg,0.83mmol) and dichloromethane (10 mL). Stir at room temperature overnight. The reaction mixture was diluted with dichloromethane, washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and then spin-dried to obtain compound 57(272mg, 100%).

LCMS:394(M+H)+,RT＝1.299mim。

EXAMPLE 46 Synthesis of Compound 58

In a dry 100mL round bottom flask was added compound 57(272mg, 0.69mmol), DHP (116mg, 1.39mmol), p-toluenesulfonic acid (67mg, 0.35mmol) and dichloromethane (10 mL). Stir at room temperature overnight. After the reaction is finished, adding potassium carbonate solution, extracting with dichloromethane, combining organic phases, drying with anhydrous sodium sulfate and spin-drying. Purification of the crude product on a column afforded compound 58(156mg, 47%). LC MS 478(M + H) +, RT 1.907mim.

EXAMPLE 47 Synthesis of Compound 59

Compound 58(70mg,0.15mmol) was dissolved in 8mL of anhydrous DMF, and trans-N, N' -dimethyl-1, 2-cyclohexanediamine (5mg,0.03mmol), CuI (2mg,0.0075mmol), and K were added to the solution in this order at room temperature₃PO₄(96mg,0.45mmol) and 58a (65mg,0.3 mmol). The mixture was purged with nitrogen 3 times, heated to 110 ℃ and stirred for 16 hours. After the reaction was complete, the solvent was spin dried to give the crude product, which was column chromatographed (dichloromethane: methanol: 40:1) to give white solid 59(50mg, 59%). LC MS:569(M + H) +, RT ═ 1.237mim.

EXAMPLE 48 Synthesis of Compound 60

Compound 59(50mg,0.088mmol) was dissolved in 10mL of dichloromethane, and trifluoroacetic acid (5mL) was added to the solution at room temperature. Stirred at room temperature for 4 hours. After the reaction was complete, the solvent was spin dried to give the crude product as a white solid 60(25mg, 59%).

¹H NMR(400MHz,MeOD-d₆)8.57(d,J＝6.8Hz,1H),8.04(d,J＝6.8Hz,2H),7.56(m,1H),7.40(m,1H),7.16(m,2H),4.11(m,2H),3.66(m,2H),3.42(m,4H),2.99(s,3H.)

EXAMPLE 49 Synthesis of Compound 62

100mg of compound 61 and Pd (dppf) Cl2/CH2Cl2(10mg) were dissolved in 3mL of methanol and 1mL of DMF and reacted at 80 ℃ under 0.5MPa of CO overnight. The reaction solution was filtered, washed and dried to obtain 65mg (65%) of a yellow liquid.

EXAMPLE 50 Synthesis of Compound 63

65mg of 62 and 19mg of hydrated lithium hydroxide were dissolved in 5mL of tetrahydrofuran and 1mL of water, and stirred at room temperature overnight. And after the reaction is completed, acidifying with 3N sulfuric acid, extracting and separating liquid with DCM, washing and drying to obtain 60mg of a product, namely the compound 63.

EXAMPLE 51 Synthesis of Compound 64

63(32mg) and 18mg of HATU were dissolved in 2mL of acetonitrile, and stirred at room temperature for 10 minutes, and 2mL of a solution of 18mg of 63a and 40mg of TEA in acetonitrile was added dropwise to the system and stirred at room temperature for about 3 hours to completion. The solution was removed by distillation under the reduced pressure, and column chromatography (DCM: MeOH ═ 30:1) gave 36mg of compound 64 (77%) as a solid.

EXAMPLE 52 Synthesis of Compound 65

36mg of 64 and 1.5mL of trifluoroacetic acid were dissolved in 3mL of DCM and stirred at room temperature overnight. The solvent was removed and preparative isolation gave 6mg (22%) of solid 65.

1H NMR(400MHz,MeOD-d4)8.33(d,1H,J＝8Hz),7.72(t,2H,J＝5.6Hz),7.66(s,1H),7.34(dd,1H,J＝8.4Hz),7.08(d,2H,J＝6.4Hz),6.95(t,1H,J＝5.2Hz),5.34(t,1H,J＝8Hz),3.93(s,6H),3.87(m,2H),3.63(m,2H),2.99(s,3H),2.17(m,2H),2.01(m,2H).

Using a similar procedure the following compounds were obtained:

EXAMPLE 53 Compound 66

6- (2, 6-difluoro-3, 5-dimethoxyphenyl) -N- (4- (4-methylpiperazin-1-yl) phenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(400MHz,DMSO-d6)13.49(s,1H),10.92(s,1H),9.78(s,1H),8.41(d,J＝8.4Hz,1H),8.06(d,J＝8.4Hz,2H),7.30(d,J＝8.4Hz,1H),7.12(m,3H),4.08(m,2H),3.93(s,6H),3.52(m,2H),3.12(m,4H),2.87(s,3H).

EXAMPLE 54 Synthesis of Compound 67

6- (2, 6-difluoro-3, 5-dimethoxyphenyl) -N- (4- ((3R, 5S) -3, 5-dimethylpiperidin-1-yl) phenyl) -1H-indazole-3-carboxamide

¹H NMR(400MHz,MeOD-d₄)8.34(d,1H,J＝8.4Hz),7.71(t,2H,J＝7.2Hz),7.66(s,1H),7.32(dd,1H,J＝8Hz),7.10(t,2H,J＝7.2Hz),6.95(t,1H,J＝8Hz),5.34(t,1H,J＝4.8Hz),3.95(s,6H),3.92(m,2H),3.52(m,2H),2.70(m,3H),1.40(d,6H,J＝6.8Hz).

EXAMPLE 55 Synthesis of Compound 68

6- (2, 6-difluoro-3, 5-dimethoxyphenyl) -N- (4- (3, 3-dimethylpiperazin-1-yl) phenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

1H NMR(400MHz,DMSO-d6)13.49(s,1H),10.91(s,1H),8.93(s,2H),8.41(d,J＝8.4Hz,1H),8.05(d,J＝8.8Hz,2H),7.30(d,J＝8.4Hz,1H),7.13(m,3H),3.93(s,6H),3.52(m,2H),3.43(m,2H),3.31(m,2H),1.37(s,6H).

Using a method similar to compound 60, the following compound was obtained:

EXAMPLE 56 Synthesis of Compound 69

4- (4-methylpiperazin-1-yl) -N- (6- (2,3,5, 6-tetrafluorophenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) benzamide

¹H NMR(400MHz,MeOD-d6)8.57(d,J＝6.8Hz,1H),8.04(d,J＝6.8Hz,2H),7.56(m,1H),7.40(m,1H),7.16(m,2H),4.11(m,2H),3.66(m,2H),3.42(m,4H),2.99(s，3H.)

EXAMPLE 57 Synthesis of Compound 71

In a 100mL three-necked flask, compound 53(300mg, 1.1mmol), 70(208.23mg,1.3mmol), Pd (dppf)₂Cl₂(83.29mg，0.1mmol)，K₂CO₃(303.5mg, 2.2mmol) followed by 12mL 1, 4-dioxane and 3mL water, the system was heated to 90 ℃ under argon atmosphere overnight, 20mL water was added after the reaction was complete, extraction was done three times with ethyl acetate (10mL), the organic phase was dried, spun dry and column chromatography gave compound 71(83mg, yield ═ 24%).

EXAMPLE 58 Synthesis of Compound 72

Compound 71(70mg, 0.22mmol) was added to a 50mL one-neck flask, then 10mL of lcm was added and dissolved, 0.2mL of TFA was slowly dropped under an ice-water bath, and finally, the temperature was slowly raised to room temperature to react for 24 hours, and after the reaction was completed, compound 72(51mg, yield ═ 99%) was obtained by spin-drying.

EXAMPLE 59 Synthesis of Compound 73

In a 25mL three-necked flask, compound 72(70mg, 0.303mmol) was added, 2mL of 1, 4-dioxane was added and dissolved, the temperature of the system was lowered to 0 ℃ under an argon atmosphere, 30% aqueous sodium hydroxide solution (1.515mL) was slowly added dropwise, then elemental iodine (92.28mg, 0.36mmol) was added, and finally the mixture was stirred at room temperature overnight, after completion of the reaction, water was added, extraction was performed with ethyl acetate, and organic phase was concentrated by drying to obtain compound 73(56mg, yield 52%).

EXAMPLE 60 Synthesis of Compound 74

In a 50mL single-neck flask, compound 73(56mg, 0.156mmol), DHP (26.39mg, 0.314mmol), TsOH (0.3mg, 0.016mmol) and 5mL of dccm were added and dissolved, and after stirring uniformly, the mixture was heated to 60 degrees reflux for 5 hours, and after completion of LCMC detection reaction, the mixture was directly dried by spin-drying and column chromatography gave compound 74(60mg, yield 87.2%).

EXAMPLE 61 Synthesis of Compound 75

In a dry 25mL three-necked flask, compound 74(60mg, 0.136mmol), compound 58a (29.52mg, 0.136mmol), CuI (17.9mg, 0.094mmol), K3PO4(57.74mg, 0.272mmol) and ligand (1S,2S) -N1, N2-dimethylcyclohexoxane-1, 2-diamine (19.34mg, 0.136mmol) were added, followed by 10mL DMF, argon blanketed and heated to 130 ℃ for 15h reaction, after which 20mL water was added, extracted with ethyl acetate, the organic phase was spin dried and column chromatographed to give compound 75(60mg, yield ═ 69.1%).

EXAMPLE 62 Synthesis of Compound 76

75(60mg, 0.1mmol) was added to a dry 25mL round bottom flask, 10mL dichloromethane was added and dissolved with stirring, 0.5mL of tfa was slowly added dropwise at room temperature, the mixture was reacted at room temperature for 18 hours, and compound 76(6mg, yield 11.8%) LC MS:449(M + H) +, RT 0.87mim was prepared by reverse phase.

¹H NMR(400MHz,d-DMSO)13.49(s,1H),10.93(s,1H),10.07(br,1H),8.42(d,J＝6.8Hz,1H),8.06(d,J＝6.8Hz,2H),7.60(m,1H),7.27-7.35(m,3H),7.13(d,J＝6.8Hz,2H),3.50-3.53(m,4H),3.14-3.17(m,4H),3.42(m,4H),2.51(s,3H.)

EXAMPLE 63 Synthesis of Compound 78

38(690mg, 2.46mmol) was dissolved in THF in a dry 250mL round-bottom flask, and after three nitrogen replacements, n-BuLi (2.3mL,3.68mmol,1.6M) was added dropwise at-78 ℃ and stirred at this temperature for 30 minutes, 77(514mg,2.94mmol) was slowly dropped into the reaction mixture at-78 ℃ and then the mixture was slowly warmed to room temperature and stirred overnight. After quenching with saturated aqueous NH4Cl solution, extraction with EA three times, drying over anhydrous sodium sulfate, spin-drying the solvent, and purification on column to obtain the target compound (450mg, 49%). LC MS 378(M + H) +.

EXAMPLE 64 Synthesis of Compound 79

78(450mg, 1.20mmol) was dissolved in DCM in a dry 100mL round-bottom flask and SOCl was added dropwise at 0 deg.C₂(1mL) was stirred at this temperature for 30 minutes, and then the temperature was gradually raised to room temperature and stirred overnight. After completion of the reaction, the solvent was spun off and purified by column chromatography to give the objective compound 79(298mg, 80%). LC MS 312(M + H)⁺。

EXAMPLE 65 Synthesis of Compound 80

In a dry 100mL round bottom flask 79(298mg, 0.96mmol) was dissolved in HOAc (10mL), then Zn powder (125mg,1.92mmol) was added to the mixture at 0 deg.C, slowly warmed to room temperature overnight. The solvent was spun off and column purified to give compound 80(199mg, 75%). LC MS:278(M + H)⁺ _。

EXAMPLE 66 Synthesis of Compound 81

In a dry 100mL round bottom flask was added compound 80(199mg, 0.72mmol), I₂(219mg, 0.86mmol), NaOH (288mg, 7.20mmol) and 1, 4-dioxane (10.0 mL). Stir at rt for 2.0 h. The reaction was extracted with dichloromethane, washed with saturated sodium thiosulfate solution, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was dried by spin-drying to give compound 81(246mg, 85%). LC MS 404(M + H)⁺.

EXAMPLE 67 Synthesis of Compound 82

A dry 250mL round bottom flask was charged with compound 81(100mg,0.20mmol),81a (47mg,0.20mmol), K₃PO₄(170mg,0.80mmol), CuI (38mg,0.20mmol),81b (113mg,0.80mmol), DMF (5mL) was replaced with nitrogen 3 times and stirred at reflux overnight. After the reaction was complete, EA (30mL x3) and water (50mLx3) were added and extractedThe separation was taken, the organic phases were combined, dried over anhydrous sodium sulfate and the solvent was dried to give compound 82(120mg, crude). LC MS 593(M + H)^+.

EXAMPLE 68 Synthesis of Compound 83

In a dry 100mL round bottom flask, add compound 82(120mg, crude) and TFA (20.0mL) and stir at 0 ℃ to room temperature for 16.0 h. The reaction solution was dried by evaporation to give 22mg of compound 83 (22%). LC MS 509(M + H)⁺N- (6- ((3, 5-dichloropyridin-4-yl) methyl) -1H-indazol-3-yl) -4- (3, 3-dimethylpiperazin-1-yl) benzamide

¹H NMR(400MHz,d-MeOD)8.26(s,1H),7.98(s,2H),7.71(d,1H,J＝6.0Hz),7.55(s,1H),7.17(s,1H),7.12(d,2H,J＝6.8Hz),7.05(d,1H,J＝6.8Hz),4.49(s,2H),3.56(s,2H),3.42(s,4H),1.49(s,6H).

Using a similar procedure the following compounds were obtained:

EXAMPLE 69 Synthesis of Compound 84

N- (6- ((2, 4-dichloropyridin-3-yl) methyl) -1H-indazol-3-yl) -4- (3, 3-dimethylpiperazin-1-yl) benzamide

¹H NMR(400MHz,d-MeOD)8.47(s,1H),7.91(d,1H,J＝6.8Hz),7.66(d,1H,J＝8.0Hz),7.12(s,1H),7.02(d,1H,J＝8.0Hz),6.97(d,1H,J＝8.0Hz),4.40(s,2H),3.46-3.49(m,2H),3.31-3.33(m,2H),3.23-3.25(m,2H),1.40(s,6H).

Using the procedure of synthesis 12, the following compounds were obtained:

EXAMPLE 70 Synthesis of Compound 85

N- (4- (4, 7-diazaspiro [2.5] oct-7-yl) phenyl) -6- (2, 6-dichloro-3, 5-dimethoxyphenyl) -1H-pyrazolo [3,4-b ] pyridine-3-carboxamide

¹H NMR(400MHz,DMSO-d6):14.45(s,1H),10.39(s,1H),9.16(s,1H),8.67(d,1H,J＝8.4Hz),7.79(d,1H,J＝9.2Hz),7.32(d,1H,J＝8.4Hz),7.07(s,1H),7.00(d,2H,J＝9.2Hz),4.00(s,6H),3.38(s,4H),3.26(s,2H),1.04-1.07(m,2H),0.94-0.97(m,2H).

Using synthesis 76 the following compound was obtained:

EXAMPLE 71 Synthesis of Compound 86

N- (6- (2, 6-dichlorophenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) -4- (4-methylpiperazin-1-yl) benzamide

1H NMR(400MHz,DMSO-d6):13.46(s,1H),10.93(s,1H),9.93(s,2H),8.43(s,1H),8.08(s,2H),7.65(s,2H),7.56(s,1H),7.01-7.27(m,4H),4.03-4.19(m,2H),3.50-3.65(m,2H),3.03-3.23(m,4H),2.89(s,3H).

Using the procedure of synthesis 65, the following compounds were obtained:

EXAMPLE 72 Synthesis of Compound 87

6- (2, 6-difluoro-3, 5-dimethoxyphenyl) -N- (4- (3, 3-dimethylpiperazin-1-yl) phenyl) -1H-indazole-3-carboxamide

¹H NMR(400MHz,DMSO-d6):13.94(s,1H),10.27(s,1H),8.98(br,2H),8.30(d,1H,J＝8.4Hz),7.79(d,1H,J＝9.2Hz),7.72(s,1H),7.72(d,1H,J＝8.4Hz),7.70(t,1H,J＝8.4Hz),7.00(d,2H,J＝9.2Hz),3.92(s,6H),3.26(br,4H),3.15(s,2H),1.39(s,6H).

Test example 1 Effect of the Compounds of the present invention on the kinase Activity of FGFR1 and FGFR2 at molecular level

1) Test method

The enzyme reaction substrate Poly (Glu, Tyr) was diluted to 20. mu.g/mL with PBS (10mM sodium phosphate buffer, 150mM NaCl, pH7.2-7.4) without potassium ions at a ratio of 4:1, 125. mu.L/well was coated on an ELISA plate, and the plate was reacted at 37 ℃ for 12-16 hours. The wells were discarded and the plate washed three times with 5 minutes each of 200. mu.L/well of T-PBS (0.1% Tween-20 in PBS). The microplate was dried in an oven at 37 ℃ for 1-2 hours.

Reaction buffer (50mM HEPES pH 7.4,50mM MgCl) was added to each well₂,0.5mM MnCl₂,0.2mM Na₃VO₄1mM DTT) was added to the sample at a final concentration of 5. mu.M. Compounds were diluted to appropriate concentrations in DMSO, 1. mu.L/well or with appropriate concentrations of DMSO (negative control wells), and reactions were initiated by the addition of recombinant protein from each kinase domain diluted in 49. mu.L of reaction buffer, with no ATP control wells required for each experiment. The reaction was carried out on a shaker (100rpm) at 37 ℃ for 1 hour. The plate was washed three times with T-PBS. 100 μ L of primary PY 99-resistant diluent was added to the reaction mixture in a shaker at 37 ℃ for 0.5 hour. The plate was washed three times with T-PBS. Adding 100 mu L/hole of IgG diluent of a second antibody horseradish peroxidase labeled goat anti-mouse, and reacting for 0.5 hour at 37 ℃ by a shaking table. The plate was washed three times with T-PBS. Adding OPD developing solution 2mg/mL (containing 0.03% H) 100 μ L/well₂O₂Diluted with 0.1M citric acid-sodium citrate buffer (pH 5.4), and reacted at 25 ℃ for 1 to 10 minutes in the absence of light. (OPD solution)Ultrasonic wave is needed during the solution, and the color developing solution needs to be prepared immediately). 2M H was added₂SO₄The reaction was stopped at 50. mu.L/well and read by a wavelength-tunable microplate reader SPECTRA MAX 190 at 490 nm.

The inhibition ratio of the sample was obtained by the following formula:

IC₅₀the values were determined by regression with a four parameter method using a microplate reader random plus software.

2) Results

Table 1 shows the IC of some of the compounds of the invention₅₀The value is obtained. Symbol + represents IC₅₀Less than 100nm, the notation ++ denotes IC₅₀Is 100nm to 500nm, and +++ represents IC₅₀Is greater than 500 nm.

TABLE 1 IC of some of the compounds of the invention₅₀Value of

The result shows that the compound can effectively inhibit the activity of various FGFR kinases at extremely low concentration (less than or equal to 100 nm).

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, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof:

in the formula,

x is selected from: H. cl, F, -CN or Me;

u, V, W, Z are each independently selected from: n or-CH-;

y is selected from: H. f, Cl or-OMe;

l is selected from:

(a)or

wherein,

j is selected from: no, -OH, halogen, -NH₂Substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted 4-7 membered heterocyclyl, substituted or unsubstituted C1-C6 alkoxy, 5-12 membered bridged ring or heterobridged ring; wherein the above groups are optionally substituted with one or more substituents selected from the group consisting of: halogen, -NH₂OH, -OH, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocyclyl, C1-C6 alkylthio; or the above radicals with C3-C6 cycloalkyl radicalsForming a spiro structure;

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

wherein Q is₁、Q₂、Q₃、Q₄Each independently selected from: n or-CH-;

B₁、B₂、B₃、B₄each independently selected from: n or-CH-.

3. The compound of claim 1, wherein R is a substituted or unsubstituted group selected from the group consisting of:

wherein,

R₁、R₂、R₃、R₄each independently selected from: H. halogen, halogen,C1-C6 alkyl, C2-C4 alkynyl, C3-C6 cycloalkyl or a five-or six-membered aryl optionally containing at least one heteroatom selected from N, O or S, or R1 and R2, and R3 and R4 are each linked to form a 3-6 membered ring;

C0-C3 alkyl is alkylene having no, or 1-3, carbon atoms;

C1-C6 alkyl is alkylene having 1 to 6 carbon atoms.

4. The compound of claim 1, wherein L is a group selected from the group consisting of:

5. The compound of claim 1, wherein the compound of formula I is selected from the group consisting of:

2)

2.2) whenQ is nothing, L isWhen said compound of formula I is selected from the group consisting of:

6. a process for the preparation of a compound of formula I according to claim 1, comprising step (a):

the definition of each group in each formula is as described above.

7. A process for the preparation of a compound of formula I according to claim 1, wherein the process further comprises step (b):

the definition of each group in each formula is as described above.

8. Use of a compound of formula I according to claim 1 for:

(b) preparing an FGFR kinase targeted inhibitor;

(c) non-therapeutically inhibiting the activity of FGFR kinase in vitro;

(d) non-therapeutically inhibiting tumor cell proliferation in vitro; and/or

9. The use of claim 8, wherein the FGFR kinase is selected from the group consisting of: FGFR1, FGFR2, FGFR3, or a combination thereof.

10. A pharmaceutical composition, comprising:

(ii) a pharmaceutically acceptable carrier.

11. A method of inhibiting FGFR kinase activity comprising the steps of:

administering to a subject in need thereof an inhibitory effective amount of a compound of formula I as defined in claim 1 or a pharmaceutically acceptable salt thereof; or

Administering to a subject in need thereof an inhibitory effective amount of the pharmaceutical composition of claim 10.