CN110256446B - Benzo-heterocycle substituted cyclopenta [4,5] pyrrolopyrazine-1-one derivatives and application thereof - Google Patents

Abstract

The invention relates to benzoheterocyclically substituted cyclopenta [4,5]]Pyrrolopyrazin-1-one derivatives, preparation methods and medical uses thereof. Specifically, the invention discloses a compound of formula (I) or a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof, and a preparation method and application thereof, wherein the definition of each group in the formula is shown in the specification and the claims.

Description

Benzo-heterocycle substituted cyclopenta [4,5] pyrrolopyrazine-1-one derivatives and application thereof

Technical Field

The invention belongs to the technical field of medicines. In particular, the invention relates to benzo-heterocycle substituted cyclopenta [4,5] pyrrolopyrazine-1-one derivatives, a preparation method thereof, application of the derivatives as BTK inhibitors, and a pharmaceutical composition prepared from the derivatives.

Background

The BTK kinase is a non-receptor tyrosine kinase in the TEC kinase family, is a key regulator of a BCR signal pathway, and has important functions on B cell maturation, proliferation and survival. BTK is over-expressed in various B cell lymphomas, and is the only clinically validated target point for effective drug development in the current TEC kinase family. Inhibition of BTK can inhibit the proliferation of a range of B cell lymphomas.

Activation of the B cell antigen receptor (BCR) signaling pathway is important for the induction and maintenance of B cell malignancies and autoimmune diseases. Bruton's tyrosine kinase (Btk) plays a key role in the BCR signal channel of hematopoietic cells and is a very good target point in the research of new therapies for lymphoma. The BTK inhibitor acts on a BCR passage to inhibit the autophosphorylation of Btk, the phosphorylation of physiological substrates PLC gamma of Btk's and the phosphorylation of downstream kinases ERK.

The BTK inhibitor acts on Chronic Lymphocytic Leukemia (CLL) cells, induces cytotoxicity, and inhibits CLL cell proliferation ability. Inhibit proliferation of BCR-activated primary B cells, and inhibit secretion of TNF alpha, IL-1 beta, IL-6, etc. in primary monocytes. The BTK inhibitor acts on a collagen-induced arthritis model, and remarkably reduces clinical arthritis symptoms such as foot swelling and joint inflammation by inhibiting B cell activity.

Only the single BTK inhibitor irrutinib is currently approved for the market, so that there is a need to develop more active, safer and more effective BTK inhibitors.

Disclosure of Invention

The invention aims to provide a compound which has a novel structure and can be used as a Btk inhibitor.

The invention provides a compound shown as a formula (I) in a first aspect, or a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof:

in the formula (I), the compound is shown in the specification,

ring A is C_6-10An aryl ring (preferably phenyl), a 5-to 6-membered monocyclic heteroaryl ring or a 4-to 6-membered unsaturated monocyclic heterocycle;

R₀is hydrogen, halogen (preferably fluorine, chlorine, bromine), C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl), halo C_1-8Alkyl (preferably halogenated C)_1-6Alkyl, more preferably halogenated C_1-3Alkyl group), C_1-8Alkoxy (preferably C)_1-6Alkoxy, more preferably C_1-3Alkoxy), halo C_1-8Alkoxy (preferably halo C)_1-6Alkoxy, more preferably halo C_1-3Alkoxy), optionally substituted C_3-8Cycloalkyl (preferably C)_3-6Cycloalkyl), C_3-8Cycloalkoxy (preferably C)_3-6Cycloalkoxy), 5-to 6-membered monocyclic heteroaryl, or-Y-L; wherein Y is a bond, - (CH)₂)_m-、-O-(CH₂)_m-or c (o); l is hydroxy, cyano-substituted C_1-3Alkyl, optionally substituted C_6-10Aryl, optionally substituted 5-to 6-membered monocyclic heteroaryl, optionally substituted 4-to 6-membered saturated monocyclic heterocycle, C_1-8Alkoxy (preferably C)_1-6Alkoxy, more preferably C_1-3Alkoxy) or NR_a0R_b0(ii) a Said optionally substituted being unsubstituted or substituted by 1,2 or 3 hydroxy, halogen, C_1-10Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl), hydroxy-substituted C_1-10Alkyl (preferably hydroxy-substituted C)_1-6Alkyl, more preferably hydroxy-substituted C_1-3Alkyl), halo C_1-8Alkyl (preferably halogenated C)_1-6Alkyl, more preferably halogenated C_1-3Alkyl, most preferably fluoro C_1-3Alkyl), 4 to 6 membered saturated mono-heterocyclic ring;

n, m are each independently 1,2 or 3; it is to be noted that n and m refer to the number of substituents on ring A, and it is understood that when R is₀By hydrogen is meant that ring a may not contain substituents, in which case the number of hydrogen atoms does not apply to the definition of n and m, for example: when ring A is a benzene ring, R₀Hydrogen is understood to mean 5 hydrogens on the phenyl ring.

Z is O or NR_a；

R₁Is halogen, C_1-10Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl) or NR_a0R_b0；

R_a、R_a0、R_b0Each independently is hydrogen or C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl groups);

R₂、R₃、R₄、R₅each independently hydrogen, halogen (preferably fluorine, chlorine, bromine), C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl), halo C_1-8Alkyl (preferably halogenated C)_1-6Alkyl, more preferably halogenated C_1-3Alkyl group), C_1-8Alkoxy (preferably C)_1-6Alkoxy, more preferably C_1-3Alkoxy) or C_3-8Cycloalkyl (preferably C)_3-6Cycloalkyl groups);

R₆、R₇each independently is hydrogen or C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl groups).

In another preferred embodiment, in formula (I), the 4 to 6 membered saturated mono-heterocyclic ring is selected from: azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, oxazolidine, piperazine, dioxolane, dioxane, morpholine, thiomorpholine-1, 1-dioxide, or tetrahydropyran.

In another preferred embodiment, in formula (I), the 5-to 6-membered monocyclic heteroaryl ring is selected from: thiophene ring, N-alkylcyclopyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, 1,2, 3-triazole ring, 1,2, 4-triazole ring, 1,2, 5-triazole ring, 1,3, 4-triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, 1,2, 3-oxadiazole ring, 1,2, 4-oxadiazole ring, 1,2, 5-oxadiazole ring, 1,3, 4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring.

In another preferred embodiment, in formula (I), the 4 to 6 membered unsaturated mono-heterocyclic ring is selected from: 2, 3-dihydro-1H-pyrrole, 2, 5-dihydrofuran, 1,2,3, 6-tetrahydropyridine, 1,2,3, 4-tetrahydropyridine, 3, 6-dihydro-2H-pyran or 3, 4-dihydro-2H-pyran.

In another preferred embodiment, in formula (I), the A ring is a benzene ring, a pyrazole ring, a pyrimidine ring, a pyridine ring or a 1,2,3, 6-tetrahydropyridine ring.

In another preferred embodiment, in formula (I), n is 1 or 2.

In another preferred embodiment, in formula (I), R₀Is hydrogen, C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl) or-Y-L; wherein Y is a bond or- (CH)₂)_m-; l is an optionally substituted 4-to 6-membered saturated mono-heterocyclic ring; said optionally substituted is unsubstituted or substituted by 1,2 or 3C_1-10Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl) substituted; m is 1.

In another preferred embodiment, in formula (I), the structure

Is a structure selected from the group consisting of:

in another preferred embodiment, the 5-to 6-membered monocyclic heteroaryl ring is selected from:

the 5-to 6-membered monocyclic heteroaryl ring is optionally substituted with 1 or 2R₀And (4) substitution.

In another preferred embodiment, the structure

Is a structure selected from the group consisting of:

in another preferred embodiment, in formula (I), the compound of formula (I) is a compound selected from the group consisting of:

in a second aspect, the present invention provides a pharmaceutical composition comprising a compound of the first aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof; and a pharmaceutically acceptable carrier or excipient.

In a third aspect, the present invention provides a compound according to the first aspect, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, or a pharmaceutical composition according to the second aspect, for use in the preparation of a kinase inhibitor medicament.

In another preferred embodiment, the kinase inhibitor is a BTK inhibitor.

In a fourth aspect, the present invention provides the use of a compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, or a pharmaceutical composition according to the second aspect of the present invention, in the manufacture of a medicament for the treatment of a B-cell mediated disease.

In a fifth aspect, the present invention provides a method of treating a B-cell mediated disease, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, or a pharmaceutical composition according to the second aspect of the present invention.

In a sixth aspect, the present invention provides a method of treating a disease mediated by B cells, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and another therapeutically active agent.

In another preferred embodiment, the disease mediated by B cells is selected from the group consisting of: a neoplastic disease, a proliferative disease, an allergic disease, an autoimmune disease, or an inflammatory disease.

In another preferred embodiment, the disease mediated by B cells is selected from the group consisting of: solid tumors, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, systemic lupus erythematosus, psoriasis, rheumatoid spondylitis, and gouty arthritis.

In another preferred embodiment, the disease mediated by B cells is a solid tumor.

In another preferred embodiment, the solid tumor is at least one selected from the group consisting of lymphoma, soft tissue sarcoma, lymphocytic lymphoma, mantle cell lymphoma, melanoma, and multiple myeloma.

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

Detailed Description

The inventors of the present invention have conducted extensive and intensive studies and have unexpectedly found that such benzoheterocycle-substituted cyclopenta [4,5] pyrrolopyrazine-1-one derivatives have high inhibitory activities against enzymes such as BTK WT and cells such as pBTKY 223. Therefore, the series of compounds are expected to be developed into medicaments for treating tumors. On this basis, the inventors have completed the present invention.

Definition of terms

As used herein, "alkyl" refers to straight and branched chain saturated aliphatic hydrocarbon groups, C_1-8The alkyl group is an alkyl group containing 1 to 8 carbon atoms, and may preferably be C_1-6Alkyl or C_1-3Alkyl, defined similarly; non-limiting examples of alkyl groups include: methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, sec-butyl group, n-pentyl group, 1-dimethylpropyl group, 1, 2-dimethylpropyl group, 2-dimethylpropyl group, 1-ethylpropyl group, 2-methylbutyl group, 3-methylbutyl group, n-hexyl group, 1-ethyl-2-methylpropyl group, 1, 2-trimethylpropyl group, 1-dimethylbutyl group, 1, 2-dimethylbutyl group, 2-dimethylbutyl group, 1, 3-dimethylbutyl group, 2-ethylbutyl group, 2-methylpentyl group, 3-methylpentyl group, 4-methylpentyl group, 2, 3-dimethylbutyl group, n-heptyl group, 2-methylhexyl group, 3-methylhexyl group, 4-methylhexyl group, 2-methylpentyl group, 3-methylhexyl group, 4-methylhexyl group, 2-dimethylpropyl group, 2-pentyl group, 2-methylpropyl group, 2-methyl-pentyl group, 3-pentyl group, 2-methyl-pentyl group, 2-pentyl group, and 3-pentyl group, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-diethylpentyl, 2, 3-dimethylpentyl, 2-dimethylhexyl, 2, 3-dimethylhexyl, 2, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-dimethylhexyl, 2-ethylhexyl, 2-ethylpentyl, 2-dimethylpentyl, 2-ethylhexyl, 3-dimethylpentyl, 2-ethylhexyl, 2-dimethylpentyl, 2-hexyl, 2-dimethylhexyl, 2-hexyl, and the like, N-decyl, 3-diethylhexyl, 2-diethylhexyl, and various branched isomers thereof.

As used herein, "cycloalkyl" refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group, ")"C_3-8Cycloalkyl "refers to a cyclic hydrocarbon group containing 3 to 8 carbon atoms, and may preferably be C_3-6Cycloalkyl, defined similarly; non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like, with cyclopropyl, cyclopentyl, cyclohexenyl being preferred.

As used herein, "C" is_1-8Alkoxy means-O- (C)_1-8Alkyl) wherein alkyl is as defined above. Preferably C_1-6Alkoxy, more preferably C_1-3An alkoxy group. Non-limiting examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentoxy, and the like.

As used herein, "C" is_3-8Cycloalkoxy means-O- (C)_3-8Cycloalkyl), wherein cycloalkyl is as defined above. Preferably C_3-6A cycloalkoxy group. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

As used herein, "C" is_6-10Aryl "refers to an all-carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, and refers to aryl groups containing 6 to 10 carbon atoms; phenyl and naphthyl are preferred, with phenyl being most preferred.

As used herein, "a bond" means that the two groups connected by it are linked by a covalent bond.

As used herein, "halogen" refers to fluorine, chlorine, bromine or iodine.

As used herein, "halo" refers to a group in which one or more (e.g., 1,2,3,4, or 5) hydrogens are replaced with a halogen.

For example, "halo C_1-8Alkyl "means an alkyl group substituted with one or more (e.g., 1,2,3,4, or 5) halogens, wherein alkyl is as defined above. Is selected from halo C_1-6Alkyl, more preferably halogenated C_1-3An alkyl group. Halogen substituted C_1-8Examples of alkyl groups include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1, 2-dichloroethyl, trichloroethyl, monobromoethylA group such as a fluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a fluoroethyl group, a difluoroethyl group, and a trifluoroethyl group.

Also for example, "halo C_1-8Alkoxy "means an alkoxy group substituted with one or more (e.g., 1,2,3,4, or 5) halogens, wherein the alkoxy group is as defined above. Preferably a halogen atom_1-6Alkoxy, more preferably halo C_1-3An alkoxy group. Halogen substituted C_1-8Examples of alkoxy groups include, but are not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, and the like.

Also for example, "halo C_3-8Cycloalkyl "refers to a cycloalkyl group substituted with one or more (e.g., 1,2,3,4, or 5) halogens, wherein cycloalkyl is as defined above. Preferably a halogen atom_3-6A cycloalkyl group. Halogen substituted C_3-8Examples of cycloalkyl groups include, but are not limited to, trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, and the like.

As used herein, "amino" refers to NH₂"cyano" means CN, "nitro" means NO₂"benzyl" means-CH₂-phenyl, "oxo" means ═ O, "carboxy" means-c (O) OH, "acetyl" means-c (O) CH₃And "hydroxymethyl" means-CH₂OH, "hydroxyethyl" means-CH₂CH₂OH or-CHOHCH₃"hydroxy" means-OH, and "thiol" means SH.

As used herein, "heteroaryl ring" is used interchangeably with "heteroaryl" and refers to a monocyclic heteroaryl group having 5 to 10 ring atoms, preferably 5 or 6 membered or a bicyclic heteroaryl group having 8 to 10 membered ring atoms; 6, 10 or 14 pi electrons are shared in the ring array; and a group having 1 to 5 hetero atoms in addition to carbon atoms. "heteroatom" means nitrogen, oxygen or sulfur.

As used herein, "4 to 6 membered saturated monocyclic heterocycle" means that 1,2 or 3 carbon atoms in the 4 to 6 membered monocyclic ring are substituted by a substituent selected from nitrogen, oxygen or S (O)_t(wherein t is an integer from 0 to 2) but excludes the ring moiety of-O-O-, -O-S-, or-S-S-, the remaining ring atoms being carbon; preferably 5 to 6 membered. Examples of 4-to 6-membered saturated monoheterocycles include (but are not limited to)) Propylene oxide, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, pyrroline, oxazolidine, piperazine, dioxolane, dioxane, morpholine, thiomorpholine-1, 1-dioxide, tetrahydropyran, and the like.

As used herein, "unsaturated" refers to a pi-electron system that contains one or more unsaturated bonds but does not have a complete conjugation.

As used herein, "4 to 6 membered unsaturated mono-heterocyclic" includes (but is not limited to): 2, 3-dihydro-1H-pyrrole, 2, 5-dihydrofuran, 1,2,3, 6-tetrahydropyridine, 1,2,3, 4-tetrahydropyridine, 3, 6-dihydro-2H-pyran, 3, 4-dihydro-2H-pyran, and the like.

As used herein, "5-to 6-membered monocyclic heteroaryl ring" refers to a monocyclic heteroaryl ring containing 5 to 6 ring atoms, including for example (but not limited to): thiophene ring, N-alkylcyclopyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, 1,2, 3-triazole ring, 1,2, 4-triazole ring, 1,2, 5-triazole ring, 1,3, 4-triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, 1,2, 3-oxadiazole ring, 1,2, 4-oxadiazole ring, 1,2, 5-oxadiazole ring, 1,3, 4-oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring, and the like.

As used herein, "hydroxy-substituted C_1-10Alkyl "means that one or more hydrogens on the alkyl group are replaced with a hydroxy group, including C substituted with a hydroxy group_1-10Alkyl, dihydroxy-substituted C_1-10Alkyl, trihydroxy-substituted C_1-10Alkyl, and the like, examples include, but are not limited to, hydroxymethyl, hydroxyethyl, 2-hydroxy-2-methylbutyl, 2-hydroxy-butyl, and the like.

As used herein, "cyano-substituted C_1-3Alkyl "means that one or more hydrogens on the alkyl group are replaced with a cyano group, including C substituted with a cyano group_1-3Alkyl, dicyano substituted C_1-3Alkyl and the like, examples include, but are not limited to, cyanomethyl, cyanoethyl and the like.

As used herein, "substituted" refers to one or more hydrogen atoms in the group, preferably 1 to 5 hydrogen atoms are substituted independently of each other with a corresponding number of substituents, more preferably 1 to 3 hydrogen atoms are substituted independently of each other with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.

As used herein, any of the above groups may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 (more preferably 1 to 3) or less groups independently selected from halogen, -O (CH)₂)_pOC_1-8Alkyl, -O (CH)₂)_pOH、-(CH₂)_pOC_1-8Alkyl, 4-to 6-membered saturated monocyclic heterocycle, C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl group), C_3-8Cycloalkyl (preferably C)_3-6Cycloalkyl), halo C_1-8Alkyl (preferably halogenated C)_1-6Alkyl, more preferably halogenated C_1-3Alkyl), halo C_3-8Cycloalkyl (preferably halo C)_3-6Cycloalkyl), hydroxy-substituted C_1-8Alkyl (preferably hydroxy-substituted C)_1-6Alkyl, more preferably hydroxy-substituted C_1-3Alkyl), hydroxymethyl, hydroxyethyl, hydroxy, carboxyl, NR_a0R_b0、-C(O)OC_1-6Alkyl, acetyl, C_1-8Alkoxy (preferably C)_1-6Alkoxy, more preferably C_1-3Alkoxy group), C_1-8Alkoxy-substituted C_1-8Alkyl (preferably C)_1-6Alkoxy-substituted C_1-6Alkyl, more preferably C_1-3Alkoxy-substituted C_1-3Alkyl), halo C_1-8Alkoxy (preferably halo C)_1-6Alkoxy, more preferably halo C_1-3Alkoxy), -SO₂C_1-8Alkyl (preferably-SO)₂C_1-6Alkyl, more preferably-SO₂C_1-3Alkyl group), C_6-10Aryl (preferably phenyl), 5-to 6-membered monocyclic heteroaryl, or-Y-L; wherein Y is (CH)₂)_qOr C (O); l is a 4 to 6 membered saturated monocyclic heterocycle or a 5 to 6 membered monocyclic heteroaryl ring; p, q are each independently 1,2 or 3; r_a0、R_b0Each independently of the others is hydrogen, acetyl, C_1-8Alkyl (preferably C)_1-6Alkyl, more preferably C_1-3Alkyl group), C_1-8Alkoxy-substituted C_1-8Alkyl (preferably C)_1-6Alkoxy-substituted C_1-6Alkyl, more preferably C_1-3Alkoxy-substituted C_1-3Alkyl groups).

The various substituent groups described herein above may themselves be substituted with groups described herein.

When the 4-to 6-membered saturated mono-heterocyclic ring described herein is substituted, the positions of the substituents may be at their possible chemical positions, and representative substitution of exemplary mono-heterocyclic rings are as follows:

wherein "Sub" represents the various types of substituents described herein;

representing a connection to another atom.

When a 4-to 6-membered saturated mono-heterocyclic ring according to the present invention is a substituent, it may itself be substituted or substituted with 1,2 or 3 substituents selected from the group consisting of: halogen, hydroxy, C_1-3Alkyl, O ═ NR_a0R_b0Hydroxymethyl, hydroxyethyl, carboxy, -C (O) OC_1-3Alkyl, acetyl, halo C_1-3Alkyl radical, C_1-3Alkoxy radical, C_3-6Cycloalkyl, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, oxazolidine, piperazine, dioxolane, dioxaneOxahexacyclic, morpholine, thiomorpholine-1, 1-dioxide, tetrahydropyran, thiophene ring, N-alkylpyrrole ring, furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, pyrimidine ring, pyrazine ring; wherein R is_a0、R_b0Each independently is hydrogen or C_1-3An alkyl group.

As used herein, "therapeutically effective amount" refers to an amount of a compound of the invention that will elicit the biological or medical response of an individual, e.g., decrease or inhibit enzyme or protein activity or ameliorate symptoms, alleviate a condition, slow or delay disease progression or prevent disease, etc.

As used herein, "pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating material or auxiliary formulation or any type of adjuvant that is compatible with the patient, preferably a mammal, more preferably a human, and that is suitable for delivering an active agent to a target site without terminating the activity of the agent.

As used herein, "patient" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs, and humans.

As used herein, "treating" or "treatment" refers to alleviating, delaying progression, attenuating, preventing, or maintaining an existing disease or disorder (e.g., cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent one or more symptoms of the disease or disorder.

Preparation method

The compounds of formula (I) of the present invention can be readily prepared by various synthetic procedures, which are well known to those skilled in the art, according to the structure of the specific compound, with reference to the exemplary preparation methods in the following examples. Reagents and raw material compounds used in the preparation process are commercially available or can be prepared by a person skilled in the art according to different designed compound structures by a known method.

Compared with the prior art, the invention has the main advantages that:

provides a series of benzo-heterocycle substituted cyclopenta [4,5] pyrrolopyrazine-1-ketone derivatives with novel structures, which have higher inhibitory activity on BTK WT and other enzymes and pBTKY 223 and other cells and can be used as medicaments for treating tumors.

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 according to conventional conditions or according to conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight. Unless otherwise defined, terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the present invention.

As used herein, DMB is 2, 4-dimethoxybenzyl, THF is tetrahydrofuran, EA is ethyl acetate, PE is petroleum ether, Ac₂O is acetic anhydride, NBS is N-bromosuccinimide, DCM is dichloromethane, AIBN is azobisisobutyronitrile, Pd (dppf) Cl₂Is 1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride, TFA trifluoroacetic acid, TBSCl tert-butyldimethylchlorosilane, NCS N-chlorosuccinimide, DHP dihydrotetrahydropyran, LiAlH₄Is lithium aluminum hydride, PMB is p-methoxybenzyl, LiHMDS is lithium bis (trimethylsilyl) amide, Pd₂(dba)₃Is tris (dibenzylideneacetone) dipalladium, RuPhos is 2-dicyclohexylphosphonium-2 ',6' -diisopropoxy-1, 1' -biphenyl, DMAP is 4-dimethylaminopyridine, THP is tetrahydropyran, N-BuLi is N-butyllithium, TMsOTf is trimethylsilyl trifluoromethanesulfonate, TEABAC is triethylbenzylammonium chloride, HATU is 2- (7-azobenzotriazol) -N, N, N ', N ' -tetramethylurea hexafluorophosphate, DMF is dimethylformamide, DMSO is dimethyl sulfoxide, DIEA is N, N-diisopropylethylamine, BINAP is (2R,3S) -2,2' -bisdiphenylphosphino-1, 1' -binaphthyl.

As used herein, room temperature means about 20-25 ℃.

Preparation of intermediate a

Step 1: compound a-1(5 g, 14.6 mmol) was dissolved in 100ml of a mixed solvent of dichloromethane and methanol (volume ratio 1/1), and sodium borohydride (1.11 g, 29.2 mmol) was added in portions in an ice bath, and after the addition was completed, the ice bath was removed and stirred at room temperature for 3 hours. And LCMS detection shows that the raw materials are completely converted. The reaction mixture was concentrated, and the remaining oil was diluted with ethyl acetate, washed twice with water, washed once with saturated brine, then dried over anhydrous sodium sulfate, filtered and concentrated to give a-2(5 g, yield: 100%) as a colorless oil.

Step 2: compound a-2(5 g, 14.5 mmol) was dissolved in 100ml of dichloromethane, triethylamine (4.39 g, 43.5 mmol) was added, then a dichloromethane solution of acetyl chloride (2.26 g, 29 mmol) was added dropwise under cooling in an ice bath, and after completion of the addition, the reaction solution was naturally warmed to room temperature and stirred overnight. The reaction was washed with water directly, dried over anhydrous sodium sulfate, filtered, concentrated, and the remaining oil was purified by column chromatography (dichloromethane/methanol ═ 30/1) to give a-3(5 g, yield 89.3%) as a pale yellow oil.

And step 3: to a 100mL round bottom flask was added compound a-3(1 g, 2.58 mmol), bis pinacol boronate (1.31 g, 5.16 mmol), Pd (dppf) Cl₂(94 mg, 0.13 mmol), X-Phos (124 mg,0.26mmol), potassium acetate (0.76 g, 7.74 mmol) and dioxane (35ml) were heated to 100 ℃ under nitrogen, stirred overnight, cooled to room temperature, filtered and concentrated, the remaining oil was diluted with 150ml ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give a black oil a (yield)>100%, crude was used directly in the next step).

Example 12 preparation of 4- (3-amino-7- (5- (4-methylpiperazin-1-yl) pyridin-2-yl) -1H-indazol-5-yl) -3- (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-2, 3,4,6,7, 8-hexahydro-1H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1-one (Z-1)

Step 1: pd is added₂(dba)₃(229mg,0.25mmol), Xantphos (290mg,0.5mmol) and sodium tert-butoxide (2.88g,30mmol) were added to a solution of 2-bromo-5-iodopyridine (3.41g,12mmol) and 1-methylpiperazine (1.0g,10mmol) in toluene (50ml), heated to 60 ℃ under argon and the reaction stirred for 24 hours. After the reaction solution is cooled to room temperature, the solid is filtered, the filter cake is washed by ethyl acetate, the filtrate is decompressed and concentrated to obtain a crude product, and the crude product is subjected to Combi-flash column chromatography [ DCM: MeOH ═ 100: 0-90: 10]After purification, the compound 1- (6-bromopyridin-3-yl) -4-methylpiperazine (2.68g, 79.3%) was obtained. M⁺＝256.0[M+1]⁺；

Step 2: a solution of compound B (1.3g,5.08mmol) in tetrahydrofuran (20ml) was cooled to-78 deg.C under argon and n-butyllithium (2.5M,3ml,7.62mmol) was slowly added dropwise. After the addition was complete, stirring was continued for 10 minutes and tributyltin chloride (2.48g,7.62mmol) was added. Slowly heating the reaction solution to room temperature, adding potassium fluoride to quench the reaction, and performing Combi-flash column chromatography on the crude product after decompression and concentration [ DCM: MeOH ═ 100: 0-95: 5]After purification, Compound Z-1-1(1.03g, 45%) was obtained. M⁺＝468.2[M+1]⁺；

And step 3: compound Z-1-1(348mg,0.75mmol) and 5-bromo-7-iodo-1H-indazol-3-amine (252mg,0.75mmol) were dissolved in 5ml dioxane, tetrakistriphenylphosphine palladium (87mg,0.75mmol) and potassium fluoride (130mg,2.24mmol) were added, and the mixture was heated to 140 ℃ in a microwave reactor under argon atmosphere and reacted for 60 minutes. After cooling to room temperature, the reaction solution was poured into water, extracted with ethyl acetate (3 × 20ml), the organic phases were combined, washed with 30ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product, which was purified by Combi-flash column chromatography [ DCM: MeOH ═ 100: 0-90: 10, 0.1% Et₃N]After purification, Compound Z-1-2(140mg, 40.6%) was obtained. M⁺＝389.0[M+1]⁺；

And 4, step 4: the compound Z-1-2(100mg,0.26mmol) and the compound a (10)3mg,0.26mmol) was dissolved in a mixed solvent of 4ml of acetonitrile and 2ml of water, PdCl was added₂(dppf) (19mg,0.026mmol), potassium phosphate (110mg,0.52mmol) and sodium acetate (71mg,0.52mmol) were heated to 110 ℃ in a microwave reactor under argon protection for 20 minutes. After the reaction solution was cooled to room temperature, it was diluted with 30ml of ethyl acetate, washed with 10ml of water and 10ml of saturated saline in this order, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by Combi-flash column chromatography [ DCM: MeOH ═ 100: 0-90: 10, 0.1% Et₃N]Purification gave Compound Z-1-3(140mg, 58.6%). M⁺＝660.3[M+1]⁺；

And 5: reacting LiOH₂O (30mg,0.71mmol) was added to a mixed solution of the compound Z-1-3(120mg,0.182mmol) in tetrahydrofuran (4ml), water (2ml) and isopropanol (2ml), and stirred at room temperature for two hours. The reaction solution was diluted with 30ml of ethyl acetate, washed with 20ml of water, washed with 20ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was purified by preparative liquid phase to give compound Z-1(35mg, 26.7%). M⁺＝618.2[M+1]⁺；¹H NMR(DMSO-d₆,400MHz)：＝11.65(s,1H),8.47(d,J＝4.9Hz,1H),8.41(d,J＝2.9Hz,1H),8.06(d,J＝1.2Hz,1H),7.94(d,J＝9.0Hz,1H),7.85(d,J＝1.2Hz,1H),7.45(dd,J＝9.0,2.9Hz,1H),7.40(d,J＝5.1Hz,1H),6.53(s,1H),5.50(br.s.,2H),4.96-5.03(m,1H),4.31-4.47(m,2H),4.10-4.30(m,3H),3.83(d,J＝7.6Hz,1H),3.27(br.s.,4H),2.51-2.60(m,2H),2.49(br.s.,4H),2.39(s,2H),2.22(s,3H),1.18ppm(br.s.,6H).

Example 22 preparation of- (3- (hydroxymethyl) -4- (7- (1-methyl-1H-pyrazol-4-yl) -3- (methylamino) -1H-indazol-5-yl) pyridin-2-yl) -7-, 7-dimethyl-2, 3,4,6,7, 8-hexahydro-1H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1-one (Z-2)

Step 1: 5-bromo-2-fluorobenzoic acid (8.68 g,40 mmol) was dissolved in 70ml of concentrated sulfuric acid, elemental iodine (10.16 g,40 mmol) was added in portions under cooling in an ice bath, the reaction solution was stirred at room temperature for 5 hours, the reaction solution was slowly poured onto crushed ice, a red solid was gradually precipitated, filtered, and dried under vacuum to give Z-2-1 as a red solid (11 g, yield 79.7%).

Step 2: in a round-bottomed flask, compound Z-2-1(3.45 g,10mmol), 25% methylamine aqueous solution (4ml), EDCI (3.84 g, 20 mmol), HOBT (2.7 g, 20 mmol), DIPEA (3.87 g,30mmol) and DMF (30ml) were added, the reaction solution was stirred overnight at room temperature, diluted with ethyl acetate, washed three times with water, washed with saturated sodium chloride, dried and concentrated, and the remaining oil was purified by column chromatography (petroleum ether/ethyl acetate: 4/1-2/1) to give Z-2-2(2.68 g,7.6 mmol) as a yellow solid.

And step 3: compound Z-2-2(0.5 g, 1.4 mmol) was dissolved in toluene (15ml), lawson's reagent (0.62 g, 1.54 mmol) was added, the reaction solution was warmed to 105 degrees, stirred for 4 hours, cooled to room temperature, concentrated, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 3/1) to give Z-2-3(0.45 g, yield: 86%) as a yellow solid.

And 4, step 4: compound Z-2-3(0.3 g, 0.8 mmol) was dissolved in DMSO (6ml), hydrazine hydrate (0.5ml) was added, the reaction was warmed to 120 degrees, stirred for 2 hours, cooled to room temperature, diluted with ethyl acetate, washed with water three times, washed with saturated sodium chloride, dried and concentrated, and the remaining oil was purified by column chromatography (petroleum ether/ethyl acetate 4/1-2/1) to give Z-2-4(0.15 g, yield 53.6%) as a yellow solid.

And 5: the compound Z-2-4(0.15 g, 0.43 mmol), 1-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (104 mg,0.5mmol), Pd (pddf) Cl₂(16 mg, 0.0215 mmol), potassium carbonate (115 mg, 0.86 mmol) were mixed in dioxane (15ml), warmed to 70 ℃ overnight under nitrogen, cooled to room temperature, filtered, diluted with ethyl acetate, washed with water, washed with saturated sodium chloride, dried and concentrated, and the remaining oil was purified by column chromatography (petroleum ether/ethyl acetate: 2/1-1/2) to give Z-2-5 as a yellow oil (90 mg, yield: 68.7%).

Step 6: compound Z-2-5(104 mg,0.34mmol), compound a (135mg,0.34mmol), pd (dppf) Cl2(12 mg, 17 mmol), sodium acetate (56 mg,0.68mmol), potassium phosphate trihydrate (181 mg,0.68mmol), and dioxane (15ml) were mixed in a microwave tube, stirred under microwave conditions (100 degrees) for 20 minutes, cooled to room temperature, diluted with ethyl acetate, washed with water, washed with saturated brine, dried, and concentrated, and the remaining oil was purified by column chromatography (dichloromethane/methanol ═ 30/1) to give Z-2-6(146 mg, 60% yield) as a yellow oil.

And 7: compound Z-2-6(197 mg,0.34mmol) was dissolved in iPA/THF/H2O ═ 8ml/4ml/4ml, lithium hydroxide monohydrate (65 mg, 1.7 mmol) was added, stirred at room temperature for 1.5 hours, diluted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated, and the residue was purified by HPLC to give Z-2(8.5 mg) as a pale yellow solid.¹H NMR(400MHz,DMSO-d₆)δ11.52(s,1H),8.45(dd,J＝5.0,1.2Hz,1H),8.29(d,J＝5.5Hz,1H),7.98(d,J＝0.9Hz,1H),7.74–7.65(m,2H),7.35(dd,J＝5.1,2.5Hz,1H),6.52(s,1H),6.06(d,J＝5.4Hz,1H),4.92(dt,J＝6.0,3.4Hz,1H),4.38(t,J＝5.1Hz,2H),4.19(m,3H),3.91–3.79(m,4H),2.87(d,J＝5.0Hz,3H),2.54(d,J＝8.2Hz,2H),2.39(s,2H),1.19(s,6H).

EXAMPLE 32 preparation of 4- (3-amino-7- (1-methyl-1H-pyrazol-4-yl) benzo [ d ] isoxazol-5-yl) -3- (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-2, 3,4,6,7, 8-hexahydro-1H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1-one (Z-3)

Step 1: the starting material acetylhydroxylamine (222 mg, 2.95 mmol) was dissolved in DMF, t-BuOK (330 mg, 2.95 mmol) was added, the temperature was raised to 30 degrees, stirring was performed for 1 hour, 5-bromo-2-fluoro-3-iodobenzonitrile (800 mg, 2.46 mmol) was added, the reaction solution was stirred overnight, diluted with ethyl acetate, washed three times with water, washed with saturated sodium chloride, dried and concentrated, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 2/1-1/1) to give Z-3-1 as a pale yellow solid (390 mg, yield: 47%).

Step 2: for detailed operation reference is made to step 5 of the final product Z-2 operation.

And step 3: for detailed operation reference is made to step 6 of the final product Z-2 operation.

And 4, step 4: for detailed operation reference is made to step 7 of the final product Z-2 operation.¹H NMR(400MHz,DMSO-d₆)δ8.53(d,J＝5.0Hz,1H),8.38(s,1H),8.07(s,1H),7.98(d,J＝1.7Hz,1H),7.79(d,J＝1.6Hz,1H),7.40(d,J＝5.0Hz,1H),6.57-6.52(m,3H),4.94(t,J＝5.2Hz,1H),4.39(d,J＝5.2Hz,2H),4.30–4.20(m,3H),3.94(s,3H),3.87(d,J＝10.4Hz,1H),2.58(d,J＝8.3Hz,2H),2.43(s,2H),1.22(s,6H).

Example 42 preparation of (3- (hydroxymethyl) -4- (3-methyl-7- (1-methyl-1H-pyrazol-4-yl) -1H-indazol-5-yl) pyridin-2-yl) -7, 7-dimethyl-2, 3,4,6,7, 8-hexahydro-1H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1-one (Z-4)

Step 1: iodine (2.54g,10.0mmol) and silver sulfate (3.12g,10.0mmol) were added to a solution of 4-bromo-2-ethylaniline (2.0g,10.0mmol) in ethanol (30ml), and the mixture was stirred at room temperature for 3 hours. The solid was removed by filtration, the filtrate was concentrated and then dissolved in 50ml of dichloromethane, washed with aqueous sodium hydroxide solution (1N, 30ml), saturated brine (30ml), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by Combi-flash column chromatography [ PE: EA is 100: 0-95: 5]After purification, the compound 4-bromo-2-ethyl-6-iodoaniline (1.97g, 60.4%) was obtained. M⁺＝328.0[M+1]⁺；¹H NMR(400MHz,DMSO-d6)7.53(d,J＝2.4Hz,1H),7.08(d,J＝2.4Hz,1H),5.02(S,2H),2.45-2.51(m,2H),1.07(t,J＝7.6Hz,3H).

Step 2: sodium nitrite (54mg,0.78mmol) was added to a solution of 4-bromo-2-ethyl-6-iodoaniline (230mg,0.71mmol) in acetic acid (5ml), the reaction was stirred at room temperature for 3 hours, and the reaction was concentrated under reduced pressure to give a crude product, which was purified by Combi-flash column chromatography [ DCM: MeOH 99: 1]Purification gave compound Z-4-1(156mg, 65.5%). M⁺＝328.0[M+1]⁺；¹H NMR(400MHz,DMSO-d6)12.98(s,1H),7.96(d,J＝1.6Hz,1H),7.80(d,J＝1.6Hz,1H),2.42(s,3H)

And step 3: PdCl₂(dppf) (92mg,0.125mmol) and potassium carbonate (345mg,2.5mmol) were added to a mixed solution of compound Z-4-1(420mg,1.25mmol) and 1-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (260mg,1.25mmol) in 8ml dioxane and 2ml water. Stir vigorously at 65 ℃ for 7 hours under nitrogen. After the reaction is finished, filtering, decompressing and concentrating the filtrate to obtain a crude product, and performing Combi-flash column chromatography [ DCM: MeOH ═ 100: 0-95: 5]Purification gave Compound Z-4-2(250mg, 68.7%). M⁺＝291.0[M+1]⁺；

And 4, step 4: PdCl₂(dppf) (50mg,0.069mmol), potassium phosphate (291mg,1.37mmol) and sodium acetate (187mg,1.37mmol) were added to a mixed solution of compound Z-4-2(200mg,0.68mmol) and compound a (273mg,0.68mmol) in 5ml of acetonitrile and 2ml of water. The reaction was carried out in a microwave reactor at 110 ℃ for 15 minutes under a nitrogen atmosphere. After the reaction is finished, filtering, decompressing and concentrating the filtrate to obtain a crude product, and performing Combi-flash column chromatography [ DCM: MeOH ═ 100: 0-95: 5]Purification gave compound Z-4-3(117mg, 30.2%).

And 5: reacting LiOH₂O (30mg,0.71mmol) was added to a mixed solution of Z-4-3(100mg,0.177mmol) in tetrahydrofuran (4ml), water (2ml) and isopropanol (2ml), and stirred at room temperature for two hours. The reaction solution was diluted with 30ml of ethyl acetate, washed with 20ml of water, washed with 20ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product, which was purified by preparative liquid phase to give compound Z-4(50mg, 54.3%). M⁺＝522.2[M+1]⁺；¹H NMR(DMSO-d₆,400MHz)：＝12.72(br.s.,1H),8.47(d,J＝4.9Hz,1H),8.35(s,1H),8.03(s,1H),7.76(d,J＝1.5Hz,1H),7.71(d,J＝1.5Hz,1H),7.42(d,J＝5.1Hz,1H),6.52(s,1H),4.99(t,J＝5.0Hz,1H),4.35(d,J＝4.9Hz,2H),4.10-4.29(m,3H),3.90(s,3H),3.79-3.87(m,1H),2.50-2.59(m,5H),2.40(s,2H),1.19ppm(s,6H).

Example 52 preparation of 4- (3-amino-7- (4- (morpholinomethyl) phenyl) -1H-indazol-5-yl) -3- (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-2, 1,3,4,6,7, 8-hexahydro-1H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1-one (Z-5)

Step 1: 5-bromo-2-fluoro-3-iodobenzonitrile (2 g, 6.1 mmol) was dissolved in THF (40ml), 85% hydrazine hydrate (1.5ml) was added, the temperature was raised to 30 ℃, stirred overnight, diluted with ethyl acetate, washed three times with water, washed with saturated sodium chloride, dried and concentrated, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 2/1-1/1) to give Z-5-1(1.9 g, yield: 91.8%) as a pale yellow solid.

Step 2: for detailed operation reference is made to step 5 of the final product Z-2 operation.

And step 3: for detailed operation reference is made to step 6 of the final product Z-2 operation.

And 4, step 4: for detailed operation reference is made to step 7 of the final product Z-2 operation.¹H NMR(400MHz,DMSO-d₆)δ11.66(s,1H),8.46(d,J＝5.0Hz,1H),7.83(d,J＝1.6Hz,1H),7.69-7.65(m,2H),7.57(d,J＝1.6Hz,1H),7.44–7.35(m,3H),5.48(s,2H),4.92(t,J＝5.1Hz,1H),4.39-4.41(m,2H),4.20-4.16(m,3H),3.85-3.87(m,1H),3.59-3.54(m,4H),3.50(s,2H),2.62–2.47(m,3H),2.38(d,J＝8.7Hz,6H),1.19(s,6H).

EXAMPLE 62 preparation of (4- (3-amino-7- (4- (morpholinomethyl) phenyl) benzo [ d ] isoxazol-5-yl) -3- (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-3-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1 (6H) -one (Z-6)

Step 1: 5-bromo-2-fluoro-3-iodobenzonitrile (920mg,12.3mmol) was dissolved in DMF (30mL), and potassium tert-butoxide (1.38g,12.3mmol) was added to the reaction solution, and the resulting reaction solution was stirred under protection of helium at room temperature overnight. Then, N-hydroxyacetamide (4g,12.3mmol) was added to the reaction solution, and the temperature was raised to 50 ℃ for reaction for 2 hours. After the reaction solution was extracted with ethyl acetate and water, the organic layer was concentrated to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain a pure product of 5-bromo-7-iodobenzo [ d ] isoxazol-3-amine (2.2g, yield 53%).

Step 2: mixing 5-bromo-7-iodobenzo [ d ]]Isoxazol-3-amine (500mg,1.5mmol), 4- (morpholinomethyl) phenylboronic acid (326mg,1.5mmol), Pd (dppf) Cl₂(110mg,0.15mmol) and K₂CO₃(622mg,4.5mmol) was placed in a microwave tube, dioxane (10mL) and water (5 drops) were added, and the reaction mixture was reacted at 80 ℃ for 40min using a microwave synthesizer. After the reaction mixture was extracted with ethyl acetate and water, the organic layer was concentrated to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain a pure compound Z-6-1(220mg, yield 38.4%).

And step 3: compound Z-6-1(220mg,0.57mmol) was dissolved in dichloromethane (20mL) and (Boc) was added to the reaction solution separately₂O (247mg,1.13mmol) and DMAP (7mg,0.057 mmol). The resulting reaction mixture was stirred at room temperature overnight. After the reaction solution was extracted with methylene chloride and water, the organic layer was concentrated to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain a pure compound Z-6-2 (280mg, yield 84%)

And 4, step 4: compound a (135mg,0.34mmol), compound Z-6-2(200mg,0.34mmol), Pd (dppf) Cl₂(25mg,0.034mmol) and K₂CO₃(141mg,1.02mmol) was placed in a microwave tube, dioxane (10mL) and water (5 drops) were added, and the reaction mixture was reacted at 110 ℃ for 30min using a microwave synthesizer. After the reaction mixture was extracted with ethyl acetate and water, the organic layer was concentrated to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain a pure compound Z-6-3 (180mg, yield 61.6%).

And 5: compound Z-6-3(170mg,0.197mmol) was dissolved in TFA (10mL), and the reaction was stirred at room temperature for 30 min. After most of TFA was removed under reduced pressure, the reaction solution was neutralized with saturated sodium bicarbonate solution, and after the reaction solution was extracted with ethyl acetate and water, the organic layer was concentrated to obtain the crude compound Z-6-4, which was used in the next step.

Step 6: compound Z-6-4(30mg,0.045mmol) was dissolved in THF (5mL) and added to LiOH (300 m)g,12.5mmol) in an aqueous solution (5 mL). The reaction mixture was stirred at room temperature for 1 hour. After most of THF was removed under reduced pressure, the reaction solution was extracted with ethyl acetate and water, and the mixture obtained after the organic layer was concentrated under reduced pressure was purified by preparative liquid chromatography to obtain the final product Z-6(8.1mg, yield 29%).¹H NMR(400MHz,DMSO-d₆)δ8.50(d,J＝5.0Hz,1H),7.98-7.96(m,2H),7.88-7.85(m,2H),7.45-7.42(m,3H),6.55-6.52(m,3H),4.96(t,J＝5.2Hz,1H),4.36(d,J＝5.2Hz,2H),4.20-4.17(m,3H),3.86-3.84(m,1H),3.55(t,J＝4.7Hz,4H),3.49(s,2H),2.62–2.51(m,3H),2.39-2.35(m,5H),1.18(s,6H).

Example 72 preparation of (4- (3-amino-7- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -1H-indazol-5-yl) -3 (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-3, 4,7, 8-tetrahydro-2H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1 (6H) -one (Z-7)

Step 1: to a 100ml round bottom flask was added 1-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1,2,3, 6-tetrahydropyridine (446 mg, 2.00 mmol), 5-bromo-7-iodo-1H-indazol-3-amine (676 mg, 2.00 mmol), Pd (dppf) Cl₂(73 mg,0.1 mmol), sodium carbonate (630 mg, 6mmol), water (5mL) and dioxane (35mL), warmed to 100 ℃ under nitrogen, stirred overnight, cooled to room temperature, filtered and concentrated, the remaining oil was diluted with 150mL ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and subjected to silica gel column chromatography to give Z-7-1(618 mg, yield: 100%) as a white solid.

Step 2: into a 100ml round-bottomed flask, the above-mentioned compound Z-7-1(618 mg, 2.00 mmol), compound a (800 mg, 2.00 mmol), Pd (dppf) Cl were added₂(73 mg,0.1 mmol), potassium carbonate (830 mg, 6mmol), water (5mL) and dioxane (35mL) under nitrogen, stirring at elevated temperature overnight, cooling to room temperature, filtering and concentrating, diluting the remaining oil with 150mL ethyl acetate, washing with water, and saturating with foodWashed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, subjected to silica gel column chromatography and then subjected to HPLC to prepare compound Z-7(200 mg, yield: 19%).¹H NMR(400MHz,DMSO-d6)δ8.44(d,J＝5.0Hz,1H),8.12(s,1H),7.73(s,1H),7.40(s,1H),7.32(d,J＝5.1Hz,1H),6.51(s,1H),6.24(s,2H),5.44(s,2H),4.89(s,1H),4.35(s,2H),4.22–4.08(m,3H),3.81(s,1H),3.12(s,2H),2.81–2.49(m,6H),2.39(s,2H),2.32(s,3H),1.18(s,6H).

Example 82 preparation of 4- (3-amino-7- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -1H-indazol-5-yl ] -3- (hydroxymethyl) pyridin-2-yl) -7, 7-dimethyl-3, 4,7, 8-tetrahydro-2H-cyclopenta [4,5] pyrrolo [1,2-a ] pyrazin-1 (6H) -one (Z-8)

Step 1: to a 100mL round bottom flask, compound Z-8-1(752 mg, 2.00 mmol), trifluoroacetic acid (5mL), and dichloromethane (5mL) were added, stirred overnight at room temperature under nitrogen, and concentrated to dryness to give Z-8-2 as an oil which was taken directly to the next step (yield: 100%).

Step 2: to a 100ml round bottom flask, compound Z-8-2(750 mg, 2.00 mmol), aqueous formaldehyde (1ml), dichloromethane (20ml), tetrahydrofuran (20ml) were added in portions, sodium borohydride acetate (1266 mg, 6.00 mmol) was added in portions, the reaction was stirred overnight, quenched with a saturated ammonium chloride solution in ice bath, extracted with dichloromethane, dried over anhydrous sodium sulfate as an organic phase, filtered, concentrated, and subjected to silica gel column chromatography to obtain Z-8-3(500 mg, yield: 86%) as a white solid.

And step 3: to a 100ml round bottom flask was added compound Z-8-3(500 mg, 1.72 mmol), 5-bromo-7-iodo-1H-indazol-3-amine (580 mg, 1.72 mmol), Pd (dppf) Cl₂(73 mg,0.1 mmol), sodium carbonate (630 mg, 6mmol), water (5mL) and dioxane (35mL) under nitrogen, heating to 100 ℃, stirring overnight, cooling to room temperature, filtering and concentrating, diluting the residual oily substance with 150mL ethyl acetate, washing with water, washing with saturated salt water, drying with anhydrous sodium sulfate, filtering, concentrating, and performing silica gel column chromatography to obtain the final productTo a white solid, Z-8-4(375 mg, yield: 58%).

And 4, step 4: to a 100mL round-bottom flask was added the above compound Z-8-4(125 mg, 0.33 mmol), compound a (132 mg, 0.33 mmol), Pd (dppf) Cl₂(15 mg,0.2 mmol), potassium carbonate (830 mg, 6mmol), water (5mL) and dioxane (35mL), under nitrogen, stirred at elevated temperature under reflux overnight, cooled to room temperature, filtered and concentrated, the remaining oil was diluted with 150mL ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and then subjected to silica gel column chromatography followed by HPLC to give compound Z-8(100 mg, yield: 50%).¹H NMR(400MHz,DMSO-d6)δ11.50(s,1H),8.45(d,J＝5.0Hz,1H),8.39(s,1H),8.00(s,1H),7.71(s,2H),7.36(d,J＝5.1Hz,1H),6.51(d,J＝5.2Hz,1H),5.46(s,2H),4.96–4.85(m,1H),4.37(d,J＝6.2Hz,2H),4.27–4.07(m,4H),3.85(d,J＝22.2Hz,1H),2.85(d,J＝6.0Hz,2H),2.56(t,J＝11.9Hz,2H),2.39(s,2H),2.19(s,3H),2.06–1.96(m,5H),1.19(s,6H).

Example 9

Compound Z-9 was prepared according to the method of example 1.¹H NMR(400MHz,DMSO-d6)δ11.70(s,1H),8.61–8.38(m,2H),8.11(s,1H),8.00(d,J＝9.0Hz,1H),7.89(s,1H),7.47(dd,J＝22.2,5.6Hz,2H),6.57(s,1H),5.54(s,2H),5.03(s,1H),4.43(s,2H),4.24(d,J＝10.6Hz,2H),3.83(d,J＝28.8Hz,4H),3.38(s,1H),3.26(s,4H),2.61(t,J＝19.8Hz,2H),2.38(d,J＝40.8Hz,2H),1.22(s,6H).

Examples 10 to 14

Compounds Z-10 to Z-14 can be prepared by the method described in example 2.

Examples 15 to 16

Compounds Z-15 to Z-16 can be prepared by the method described in example 3.

Example 17

Compound Z-17 can be prepared by the method described in example 5.¹H NMR(400MHz,DMSO-d₆)δ11.80(s,1H),8.65(d,J＝2.2Hz,1H),8.48(d,J＝5.0Hz,1H),8.20(d,J＝1.5Hz,1H),8.07(d,J＝8.3Hz,1H),7.95(d,J＝1.4Hz,1H),7.80(dd,J＝8.2,2.2Hz,1H),7.63–7.47(m,2H),7.41(d,J＝5.0Hz,1H),6.53(s,1H),5.53(s,2H),5.03–4.95(m,1H),4.39(t,J＝4.7Hz,2H),4.31–4.04(m,3H),3.86-3.84(m,1H),3.57-3.53(m,6H),2.55(d,J＝8.3Hz,2H),2.39-2.36(m,4H),1.19(d,J＝2.7Hz,6H).

Example 18

Compound Z-18 can be prepared by a method described in example 8.¹H NMR(400MHz,DMSO-d6)δ11.52(s,1H),8.54–8.32(m,2H),8.01(s,1H),7.71(s,2H),7.36(d,J＝5.0Hz,1H),6.52(s,1H),5.49(s,2H),4.99–4.86(m,1H),4.40(d,J＝12.6Hz,3H),4.30–4.06(m,3H),3.96(d,J＝11.2Hz,2H),3.82(s,1H),3.47(t,J＝9.8Hz,2H),2.54(d,J＝8.4Hz,2H),2.39(s,2H),2.00(d,J＝18.4Hz,4H),1.19(s,6H).

Biological assay

Test example-Lantha screening kinase reaction experiment method

Compounds were pre-dissolved in 100% DMSO. 10mM of drug stock was dissolved at room temperature and diluted in 8 vol% DMSO gradient to a final concentration of 10-0.005. mu.M. The reaction was initiated by adding 2.5. mu.l of the analyte solution and 2.5. mu.l of the kinase (Invitrogen PV3363) diluted in the reaction buffer to each well of a 384-well plate (Corning 3676), and diluting a mixture of Fluososci-PolyGT (Invitrogen PV3610) substrate and ATP (Invitrogen PV3227) with 5. mu.l of the reaction buffer. Where Blank wells (Blank) were used in place of kinase with reaction buffer and kinase wells (Enzyme) were not loaded with any drug. After shaking the reaction mixture at 25 ℃ for 60 minutes in the absence of light. Mu.l of Detection Solution (Invitrogen PV3528 mixed Solution with EDTA diluted with TR-FRET dilution buffer at a working concentration of 5mM EDTA and 0.2nM lanthastening Tb PY20 antibody) was added and the mixture was subjected to shake reaction at room temperature for 30 minutes. The plate was read on a VictorX5 fluorescence microplate reader (PerkinElmer) and the light absorption was measured at an excitation wavelength of 340nm, emission wavelengths of 500nm and 520 nm.

The inhibition was calculated as follows (see Invitrogen, specification PV 3363):

1.Emission rate(ER)：Coumarin Emission(520nm)/Fluorescein Emission(500nm)

2. inhibition Ratio (IR): (ER)_kinase-ER_{test compound})/(ER_kinase-ER_blank) X 100%. Half inhibitory concentration IC50 was calculated by fitting using XLFIT 5.0 software (IDBS, england). The results are shown in table 1:

inhibitory Activity of the Compounds of Table 1 on BTK WT

Test example two experimental methods for detecting HTRF by intracellular beta BTK Y223 phosphorylation

Compounds were pre-dissolved in 100% DMSO. 10mM of drug stock was dissolved at room temperature and diluted in 5 vol% DMSO gradient to a final concentration of 3-0.0014. mu.M. Ramos cells were plated at 4X 10⁵Per well density into 96-well plate, 45. mu.l each well of 1640 medium containing 10% (V/V) FBS, 5. mu.l each well of diluted analyte solution, 37 ℃ and 5% (V/V) CO₂The culture was carried out for 1 hour. Add 10. mu.l of sodium peroxovanadate diluent (serum-free 1640 dilution) to the negative control wells, 10. mu.l of noneSerum medium was incubated with shaker at 25 ℃ for 30 min. Mu.l of lysis buffer (4 Xlysis: blocking stock 25: 1) was added to each well and incubated for 30min at 25 ℃ in a shaker. After shaking at 800rpm for 1 minute, 16. mu.l of the cell lysate was added to a 384-well plate (Greiner784075), 4. mu.l of the premixed antibody solution (Phospho-BTK d2 antibody and Phospho-BTK Cryptate antibody were diluted 20-fold using the detection solution) was added, and the mixture was incubated overnight at 25 ℃ with shaking. The plates were read on a VictorX5 fluorescence microplate reader (PerkinElmer) and the light absorption was measured at 317nm excitation wavelength, 500nm emission wavelength and 520nm emission wavelength (cf. Cisbio, 63ADK017PEH instructions). Half inhibitory concentration IC50 was calculated by fitting using XLFIT 5.0 software (IDBS, england). The results are shown in table 2:

TABLE 2 inhibitory Activity of Compounds on beta BTK Y223 cells

As can be seen from tables 1 and 2, representative compounds of the present invention have high inhibitory activity against enzymes and cells.

Test example three Activity inhibition test for wild-type EGFR kinase

Reagents used in the following z-lyte test methods are all available from Invitrogen.

The inhibitory activity of the test substances against wild-type EGFR kinase (Invitrogen, PV3872) was determined by the z-lyte method.

The working concentrations of the components in the 10uL wild-type EGFR kinase reaction system were: mu.M ATP, 0.8 ng/. mu.L wild type EGFR kinase (Invitrogen, PV3872), 2. mu.M Tyr04 substrate (Invitrogen, PV 3193). The final concentration of DMSO after addition of the test substance was 2%.

A10 mM stock of drug dissolved at room temperature was diluted to a final concentration of 10-0.005. mu.M with a gradient of 4% DMSO in water. 2.5. mu.L of the test substance solution and 5. mu.L of a mixture of wild-type EGFR kinase and Tyr04 substrate diluted in the reaction buffer were added to each wellThen, 2.5. mu.L of ATP was added to initiate the reaction. Wherein the C1 well is used as a reaction buffer to replace ATP, the C2 well is not added with any drug, and the C3 well is added with a phosphorylated substrate as described in the specification. After 60 minutes of reaction at 25 ℃ with shaking and protection from light. 5 μ L of fusion Reagent B (Invitrogen, diluted with TR-FRET dilution buffer) was added and the reaction was shaken for 60 minutes at room temperature. The plates are read on a VictorX5 fluorescence microplate reader (PerkinElmer) and the light absorption at an excitation wavelength of 405nm, emission wavelengths of 450nm and 520nm is determined (e.g., C3)_520nmIndicating the reading at 520nm of the C3 pore).

The inhibition was calculated as follows (see Invitrogen, specification PV 3193):

1、ER＝Coumarin Emission(450nm)/Fluorescein Emission(520nm)

2. phosphorylation rate (1- ((ER × C3)_520nm-C3_450nm)/((C1_450nm-C3_450nm)+ER×(C3_520nm-C1_520nm))))×100％

3. Inhibition Ratio (IR) ═ 1- (phosphorylation rate of test compound)/(phosphorylation rate of C2)) × 100%

Half maximal inhibitory concentration IC was calculated by fitting XLFIT 5.0 software (IDBS, UK)₅₀。

TABLE 3 inhibitory Activity of Compounds on EGFR WT

As can be seen from table 3, representative compounds of the invention have lower inhibitory activity against wild-type EGFR kinase. Thus, the exemplary compounds of the invention have selective inhibitory activity against BTK WT kinase.

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 (16)

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

in the formula (I), the compound is shown in the specification,

ring a is phenyl, a 5-to 6-membered monocyclic heteroaryl ring, or a 4-to 6-membered unsaturated monocyclic heterocycle;

R₀is hydrogen, halogen, C_1-8Alkyl, halo C_1-8Alkyl radical, C_1-8Alkoxy, halo C_1-8Alkoxy, optionally substituted C_3-8Cycloalkyl radical, C_3-8Cycloalkoxy or-Y-L; wherein Y is a bond or- (CH)₂)_m-; l is hydroxy, cyano-substituted C_1-3Alkyl, optionally substituted 4-to 6-membered saturated monocyclic heterocycle, C_1-8Alkoxy or NR_a0R_b0(ii) a Said optionally substituted being unsubstituted or substituted by 1,2 or 3 hydroxy, halogen, C_1-10Alkyl, hydroxy substituted C_1-10Alkyl, halo C_1-8Alkyl substituted;

n, m are each independently 1,2 or 3;

z is O or NR_a；

R₁Is halogen, C_1-10Alkyl or NR_a0R_b0；

R_a、R_a0、R_b0Each independently is hydrogen or C_1-8An alkyl group;

R₂、R₃、R₄、R₅each independently is hydrogen;

R₆、R₇each independently is C_1-8An alkyl group.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), the 4-to 6-membered saturated monoheterocycle is selected from the group consisting of: azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, oxazolidine, piperazine, dioxolane, dioxane, morpholine, or tetrahydropyran.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), the 5-to 6-membered monocyclic heteroaryl ring is selected from: a thiophene ring, an N-alkyl ring pyrrole ring, a furan ring, a thiazole ring, an imidazole ring, an oxazole ring, a pyrrole ring, a pyrazole ring, a triazole ring, a tetrazole ring, an isoxazole ring, an oxadiazole ring, a thiadiazole ring, a pyridine ring, a pyridazine ring, a pyrimidine ring, or a pyrazine ring.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), the 4-to 6-membered unsaturated mono-heterocyclic ring is selected from: 2, 3-dihydro-1H-pyrrole, 2, 5-dihydrofuran, 1,2,3, 6-tetrahydropyridine, 1,2,3, 4-tetrahydropyridine, 3, 6-dihydro-2H-pyran or 3, 4-dihydro-2H-pyran.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), ring a is a benzene ring, a pyrazole ring, a pyrimidine ring, a pyridine ring, or a 1,2,3, 6-tetrahydropyridine ring.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), n is 1 or 2.

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula (I), R is₀Is hydrogen, C_1-3Alkyl or-Y-L; wherein Y is a bond or- (CH)₂)_m-; l is optionally substituted piperidine, piperazine, morpholine or tetrahydropyran; said optionally substituted piperidine, piperazine, morpholine or tetrahydropyran is unsubstituted or substituted by 1 or 2C_1-3Alkyl substituted; m is 1.

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, of formula (I) having the structure

Is any one structure selected from the following group:

9. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the 5-to 6-membered monocyclic heteroaryl ring is selected from:

the 5-to 6-membered monocyclic heteroaryl ring is optionally substituted with 1 or 2R₀And (4) substitution.

10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having structure (la)

Is any one structure selected from the following group:

11. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) is a compound selected from any one of:

12. a pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

13. Use of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12, in the manufacture of a medicament for use as a kinase inhibitor.

14. The use of claim 13, wherein the kinase inhibitor drug is a drug for the treatment of a B cell mediated disease.

15. The use according to claim 13, wherein the kinase inhibitor medicament is a medicament for the treatment of at least one of a proliferative disease, an allergic disease, an autoimmune disease, and an inflammatory disease.

16. The use according to claim 13, wherein the kinase inhibitor medicament is a medicament for the treatment of a neoplastic disease.