CN114957242A - Preparation and application of pyrido-heterocyclic compound as kinase inhibitor - Google Patents

Abstract

The invention relates to a novel Bruton's tyrosine kinase inhibitor, which is a compound containing a polyaromatic heterocyclic structure and comprises compounds shown in formulas (I) and (II) or isomers, stable isotope derivatives, hydrates, solvates, polymorphs and pharmaceutically acceptable salts thereof, and also discloses a preparation method of the compounds and application of the novel compounds to treatment or prevention of Bruton's tyrosine kinase related diseases such as Acute Lymphocytic Leukemia (ALL), Chronic Myelocytic Leukemia (CML), Mantle Cell Lymphoma (MCL), colorectal cancer, rheumatoid arthritis, organ transplant rejection resistance, psoriasis resistance, lupus erythematosus and the like.

Description

Preparation and application of pyrido-heterocyclic compound as kinase inhibitor

Technical Field

The invention belongs to the field of drug synthesis, and particularly relates to a novel Bruton's tyrosine kinase inhibitor, and a preparation method and application thereof.

Background

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

Bruton's tyrosine kinase small molecule inhibitors such as ibrutinib (ibrutinib), acartinib (acalabutinib) and zebrabrutinib (zanubutinib) are currently approved by the FDA in the united states for marketing for the treatment of Mantle Cell Lymphoma (MCL) and CLL.

Although ibrutinib, acatinib and zetinib are effective in treatment, a significant proportion of patients with clinical B-cell lymphoma are not susceptible to treatment except for a proportion of patients who develop resistance in later stages, for example, about 1/3 patients in MCL are not responsive to treatment and the response rate in DLBCL is not high. In view of the above, there remains a need in the art to develop Bruton's tyrosine kinase inhibitors that are highly active and specific.

Disclosure of Invention

In order to solve the above problems, the present invention provides a novel Bruton's tyrosine kinase inhibitor compound represented by formula (I) or a stereoisomer, stable isotope derivative, hydrate, solvate, or pharmaceutically acceptable salt thereof:

X ₁ ，X ₂ ，X ₄ can be independently selected from N, CR ¹ ；

The bonds a and b are single bonds or double bonds;

X ₃ may be independently selected from absent N, CR ¹

Ar ₁ And Ar ₂ Independently selected from a phenyl ring or a 5-6 membered heteroaromatic ring wherein said phenyl and heteroaromatic rings are optionally substituted by one or more G ¹ Substituted;

R ¹ independently selected from H, cyano, halogen, C _1-6 Alkyl, COOH, CONH2, NHCOH, CONHR2, OR ² or-NHR ² ；

R ² Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -OR ³ 、-NR ³ R ⁴ 、-C(O)NR ³ R ⁴ Wherein said alkyl, cycloalkyl OR heterocycloalkyl is optionally substituted by cyano, halogen, -OR ⁵ 、-NR ⁵ R ⁶ 、C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocycloalkyl;

u and W are independently selected from-C _0-4 Alkyl-, -CR ⁷ R ⁸ -、-C _1-2 Alkyl (R) ⁷ )(OH)-、-C(O)-、-CR ⁷ R ⁸ O-、-OCR ⁷ R ⁸ -、-SCR ⁷ R ⁸ -、-CR ⁷ R ⁸ S-、-NR ⁷ -、-NR ⁷ C(O)-、-C(O)NR ⁷ -、-NR ⁷ C(O)NR ⁸ -、-CF ₂ -、-O-、-S-、-S(O) _m -、-NR ⁷ S(O) ₂ -、-S(O) ₂ NR ⁷ -；

Y is absent or C is selected _3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spiro cyclic group, 5-12 membered spiro heterocyclic group, aromatic group or heteroaromatic group, wherein said cycloalkyl, heterocycloalkyl, spiro cyclic group, fused heterocyclic group, spiro heterocyclic group, aromatic group or heteroaromatic group is optionally substituted with one or more G ¹ Substituted;

z is independently selected from cyano, -NR ¹² CN、

Bond c is a double or triple bond;

when c is a double bond, R ^a 、R ^b And R ^c Each independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl. Wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ² Substituted;

R ^a and R ^b Or R ^b And R ^c Optionally taken together with the carbon atom to which they are attached to form a 3-6 membered ring optionally containing heteroatoms;

when the bond c is a triple bond, R ^a And R ^c Is absent, R ^b Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl by one or more G ³ Substituted;

R ¹² independently selected from H, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl, wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ⁴ Substituted;

G ¹ 、G ² 、G ³ and G ⁴ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹³ 、-OC(O)NR ¹³ R ¹⁴ 、-C(O)OR ¹³ 、-C(O)NR ¹³ R ¹⁴ 、-C(O)R ¹³ 、-NR ¹³ R ¹⁴ 、-NR ¹³ C(O)R ¹⁴ 、-NR ¹³ C(O)NR ¹⁴ R ¹⁵ 、-S(O) _m R ¹³ or-NR ¹³ S(O) _m R ¹⁴ Wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by 1 or more cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-to 10-membered heteroaryl,-OR ¹⁶ 、-OC(O)NR ¹⁶ R ¹⁷ 、-C(O)OR ¹⁶ 、-C(O)NR ¹⁶ R ¹⁷ 、-C(O)R ¹⁶ 、-NR ¹⁶ R ¹⁷ 、-NR ¹⁶ C(O)R ¹⁷ 、-NR ¹⁶ C(O)NR ¹⁷ R ¹⁸ 、-S(O) _m R ¹⁶ or-NR ¹⁶ S(O) _i R ¹⁷ Substituted with the substituent(s);

R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ 、R ⁹ 、R ¹⁰ 、R ¹¹ 、R ¹³ 、R ¹⁴ 、R ¹⁵ 、R ¹⁶ 、R ¹⁷ and R ¹⁸ Each independently selected from hydrogen, cyano, halogen, C _1-6 Alkyl radical, C _3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl; and m is 1 or 2.

Typical compounds of the invention include, but are not limited to, the following:

the invention provides a novel Bruton's tyrosine kinase inhibitor or an isomer, a hydrate, a solvate, a polymorph, a pharmaceutically acceptable salt and application of a pharmaceutically acceptable carrier in preparing the novel Bruton's tyrosine kinase inhibitor.

The pharmaceutical composition is in the form of tablets, capsules, granules, sprays or injections.

The pharmaceutically acceptable carrier is selected from one or more of a filler, a disintegrant, a binder and a lubricant. Including but not limited to any and all solvents, dispersion media, coatings, absorption delaying agents, and the like, for use in the pharmaceutical active substance application in the art.

The invention also provides application of the novel Bruton's tyrosine kinase inhibitor or isomer, hydrate, solvate, polymorph and pharmaceutically acceptable salt thereof as the Bruton's tyrosine kinase inhibitor.

Further, the protein tyrosine kinase inhibitor is Bruton's tyrosine kinase inhibitor.

Use of a novel Bruton's tyrosine kinase inhibitor or an isomer, hydrate, solvate, polymorph, pharmaceutically acceptable salt or pharmaceutical composition thereof in the manufacture of a medicament for the treatment or prevention of Bruton's tyrosine kinase related diseases.

Further, the Bruton's tyrosine kinase related diseases are selected from the group consisting of: acute Lymphocytic Leukemia (ALL), Chronic Myelocytic Leukemia (CML), Mantle Cell Lymphoma (MCL), carcinoma of large intestine, rheumatoid arthritis, organ transplant rejection, psoriasis, lupus erythematosus, etc.

It will be apparent that various other modifications, substitutions and alterations can be made in the present invention without departing from the basic technical concept of the invention as described above, according to the common technical knowledge and practice in the field.

The above-described aspects of the present invention will be further described in detail with reference to the following specific embodiments. It should not be understood that the scope of the above-described subject matter of this invention is limited to the following examples. All the technologies implemented based on the above-mentioned contents belong to the scope of the present invention.

Certain chemical terms

Unless stated to the contrary, the following terms are used in the specification and claims.

Has the following meanings and is used herein in the manner of _x-y "denotes the range of the number of carbon atoms, wherein x and y are each an integer, e.g. C _3-8 Cycloalkyl denotes cycloalkyl having 3 to 8 carbon atoms, i.e. having3. Cycloalkyl of 4,5, 6, 7 or 8 carbon atoms. It is also understood that "C" is _3-8 "also includes any subrange therein, e.g. C _3-7 、C _3-6 、C _4-7 、C _4-6 、C _5-6 And the like.

"alkyl" refers to a straight or branched chain hydrocarbyl group containing 1 to 20 carbon atoms, for example 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, and 2-ethylbutyl. The alkyl group may be substituted or unsubstituted.

"alkenyl" refers to a straight or branched chain hydrocarbyl group containing at least one carbon-carbon double bond and typically 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1, 4-pentadienyl, and 1, 4-butadienyl. The alkenyl group may be substituted or unsubstituted.

"alkynyl" refers to a straight or branched chain hydrocarbyl group containing at least one carbon-carbon triple bond and typically 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl. The alkynyl group may be substituted or unsubstituted.

"cycloalkyl" refers to a saturated cyclic hydrocarbyl substituent containing from 3 to 14 carbon ring atoms. Cycloalkyl groups may be monocyclic, typically containing from 3 to 7 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Cycloalkyl groups can alternatively be fused together in two or three rings, such as decahydronaphthyl, which can be substituted or unsubstituted.

"Heterocyclyl", "heterocycloalkyl", "heterocycle" means a stable 3-to 18-membered monovalent non-aromatic ring comprising 2 to 12 carbon atoms, 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise specified, a heterocyclyl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may contain fused, spiro or bridged ring systems, the nitrogen, carbon or sulfur of the heterocyclyl group may optionally be oxidized, the nitrogen atom may optionally be quaternized, and the heterocyclyl group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule through a single bond via a carbon or heteroatom in the ring. The heterocyclic group containing fused rings may contain one or more aromatic or heteroaromatic rings, provided that the atoms on the non-aromatic ring are attached to the rest of the molecule. For purposes of this application, a heterocyclyl group is preferably a stable 4-11 membered monovalent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably a stable 4-8 membered monovalent non-aromatic monocyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclyl groups include azepanyl, azetidinyl, decahydroisoquinolinyl, dihydrofuranyl, indolinyl, dioxolanyl, 1-dioxo-thiomorpholinyl, imidazolidinyl, imidazolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinonyl, pyranyl, pyrazolidinyl, pyrrolidinyl, quinolizinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydropyranyl and the like.

"Spiroheterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group with one atom (called the spiro atom) shared between monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O) _m (wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings has a fully conjugated electronic system, preferably 6 to 14, more preferably 7 to 10. The spirocycloalkyl groups are divided into mono-spiroheterocyclyl, di-spiroheterocyclyl or poly-spiroheterocyclyl groups, depending on the number of spiro atoms shared between the rings, preferably mono-spirocycloalkyl anda spiro cycloalkyl group. More preferably a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monocyclic group. Non-limiting examples of spiroheterocyclyl radicals include:

"fused heterocyclyl" means a 5 to 20 membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system in which one or more ring atoms are selected from nitrogen, oxygen or S (O) _m (wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

"aryl" or "aryl" refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 membered, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring.

"heteroaryl" or "heteroaryl" refers to a 5-16 membered ring system containing 1-15 carbon atoms, preferably 1-10 carbon atoms, 1-4 heteroatoms selected from nitrogen, oxygen and sulfur, at least one aromatic ring. Unless otherwise specified, heteroaryl groups may be monocyclic, bicyclic, tricyclic or tetracyclic ring systems, which may contain fused or bridged ring systems, provided that the point of attachment to the rest of the molecule is an aromatic ring atom, which may be selectively oxidized at nitrogen, carbon and sulfur atoms, and which may optionally be quaternized. For the purposes of the present invention, heteroaryl groups are preferably stable 4-11 membered monocyclic aromatic rings containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably stable 5-8 membered monocyclic aromatic rings containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heteroaryl groups include acridinyl, azepinyl, benzimidazolyl, benzindolyl, benzodioxinyl, benzodioxolyl, benzofuranonyl, benzofuranyl, benzonaphthofuranyl, benzopyranonyl, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazolyl, furanyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quininyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, and the like. In the present application, heteroaryl is preferably 5-8 membered heteroaryl comprising 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably pyridyl, pyrimidinyl, thiazolyl. The heteroaryl group may be substituted or unsubstituted.

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

"hydroxy" means-OH, and "amino" means-NH ₂ "amido" means-NHCO-, "cyano" means-CN, "nitro" means-CN, "isocyano" means-NC, and "trifluoromethyl" means-CF ₃ 。

The term "heteroatom" or "hetero", as used herein alone or as part of another ingredient, refers to atoms other than carbon and hydrogen, and is independently selected from, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin, and in embodiments where two or more heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different.

The terms "fused" or "fused ring" as used herein, alone or in combination, refer to a cyclic structure in which two or more rings share one or more bonds.

The term "spiro" or "spirocyclic" as used herein, alone or in combination, refers to a cyclic structure in which two or more rings share one or more atoms.

"optionally" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes where the event or circumstance occurs or does not occur-for example, "heterocyclic group optionally substituted with alkyl" means that alkyl may, but need not, be present, and that the description includes instances where the heterocyclic group is substituted with alkyl and instances where the heterocyclic group is not substituted with alkyl.

"substituted" means that one or more atoms, preferably 5, more preferably 1 to 3 atoms, in the group are independently substituted with a corresponding number of substituents. It goes without saying that the skilled person in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort, when the substituents are in their possible chemical positions. For example, having a free amine or hydroxyl group may be unstable in combination with a carbon atom having an unsaturated (e.g., olefinic) bond. Such substituents include, but are not limited to, hydroxy, amine, halogen, cyano, C _1-6 Alkyl radical, C _1-6 Alkoxy radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl groups, and the like.

"pharmaceutical composition" refers to a composition containing one or more compounds described herein, or a pharmaceutically acceptable salt or prodrug thereof, and other ingredients such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote administration to the organism, facilitate absorption of the active ingredient and further exert biological activity.

"isomers" refer to compounds having the same molecular formula but differing in the nature or order of their bonding of atoms or the spatial arrangement of their atoms, referred to as "isomers", and isomers differing in the spatial arrangement of their atoms, referred to as "stereoisomers". Stereoisomers include optical isomers, geometric isomers and conformational isomers. The compounds of the present invention may exist in the form of optical isomers. Depending on the configuration of the substituents around the chiral carbon atom, these optical isomers are either in the "R" or "S" configuration. Optical isomers, including enantiomers and diastereomers, and methods of preparing and separating optical isomers are known in the art.

Geometric isomers may also exist for the compounds of the present invention. The present invention contemplates various geometric isomers and mixtures thereof resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as either the Z or E configuration, substituents around cycloalkyl or heterocyclic rings are designated as either the cis or trans configuration.

The compounds of the invention may also exhibit tautomerism, such as keto-enol tautomerism.

It is to be understood that the present invention includes any tautomeric or stereoisomeric form and mixtures thereof, and is not to be limited solely to any one tautomeric or stereoisomeric form employed in the nomenclature or chemical structure of the compounds.

"isotopes" are all isotopes of atoms occurring in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into compounds of the invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as but not limited to ² H、 ³ H、 ¹³ C、 ¹⁴ C、 ¹⁵ N、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F and ³⁶ and (4) Cl. Isotopically-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples using appropriate isotopically-labeled reagents in place of non-isotopically-labeled reagents. Such compounds have a variety of potential uses, for example, as standards and reagents in the determination of biological activity. In the case of stable isotopes, such compounds have the potential to favorably alter biological, pharmacological or pharmacokinetic properties.

By "prodrug" is meant that the compounds of the present invention can be administered in the form of a prodrug. Prodrugs refer to derivatives that are converted to the biologically active compounds of the invention under physiological conditions in vivo, e.g., by oxidation, reduction, hydrolysis, and the like, each of which utilizes or proceeds without the participation of an enzyme. Examples of prodrugs are the following compounds: compounds in which the amine group in the compounds of the invention is acylated, alkylated or phosphorylated, for example eicosanoylamino, propylaminoylamino, pivaloyloxymethylamino, or in which the hydroxyl group is acylated, alkylated, phosphorylated or converted to a borate, for example acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaroyloxy, propylaminoyloxy, or in which the carboxyl group is esterified or amidated, or in which the sulfhydryl group forms a disulfide bridge with a carrier molecule, for example a peptide, which selectively delivers a drug to the target and/or to the cytosol of the cell, can be prepared from the compounds of the invention according to well-known methods.

"pharmaceutically acceptable salt" or "pharmaceutically acceptable" refers to those made from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. Where the compounds of the invention contain one or more acidic or basic groups, the invention also includes their corresponding pharmaceutically acceptable salts. Thus, the compounds of the invention containing acidic groups can be present in the form of salts and can be used according to the invention, for example as alkali metal salts, alkaline earth metal salts or as ammonium salts. More specific examples of such salts include sodium, potassium, calcium, magnesium or salts with amines or organic amines, such as primary, secondary, tertiary, cyclic amines, and the like, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purines, piperazine, piperidine, choline, caffeine, and the like, with particularly preferred organic bases being isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. The compounds of the invention containing basic groups can be present in the form of salts and can be used according to the invention in the form of their addition to inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. If the compounds of the invention contain both acidic and basic groups in the molecule, the invention also includes inner salts or betaine salts in addition to the salt forms mentioned. The respective salts are obtained by customary methods known to the person skilled in the art, for example by contacting these with organic or inorganic acids or bases in solvents or dispersants or by anion exchange or cation exchange with other salts.

Thus, when reference is made in this application to "a compound", "a compound of the invention" or "a compound of the invention", all said compound forms are included, such as prodrugs, stable isotopic derivatives, pharmaceutically acceptable salts, isomers, meso-forms, racemates, enantiomers, diastereomers and mixtures thereof.

In this context, the term "tumor" includes both benign tumors and malignant tumors (e.g., cancers).

The term "cancer" as used herein includes various malignancies in which Bruton's tyrosine kinase is involved, including, but not limited to, non-small cell lung cancer, esophageal cancer, melanoma, striated muscle garnet, cell carcinoma, multiple myeloma, breast cancer ovarian cancer, endometrial cancer, cervical cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer and liver cancer (e.g., hepatocellular carcinoma), more specifically liver cancer, gastric cancer and bladder cancer.

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

The term "polymorph" or "polymorph (phenomenon)" as used herein means that the compounds of the present invention have multiple crystal lattice forms, some of the compounds of the present invention may have more than one crystal form, and the present invention encompasses all polymorphic forms or mixtures thereof.

Intermediate compounds of the present invention and polymorphs thereof are also within the scope of the present invention.

Crystallization often results in a solvate of a compound of the present invention, and the term "solvate" as used herein refers to an association of one or more molecules of a compound of the present invention and one or more molecules of a solvent.

The solvent may be water, in which case the solvate is a hydrate. In addition, an organic solvent may be used. Thus, the compounds of the present invention may exist as hydrates, including monohydrate, dihydrate, hemihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compounds of the invention may be true solvates, but in other cases the compounds of the invention may also be present only occasionally as water or as a mixture of water with some other solvent the compounds of the invention may be reacted in a solvent or precipitated or crystallized in a solvent. Solvates of the compounds of the invention are also included within the scope of the invention.

As used herein, the term "acceptable" in reference to a formulation, composition or ingredient means that there is no lasting deleterious effect on the overall health of the subject being treated.

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

"pharmaceutically acceptable carriers" include, but are not limited to, adjuvants, carriers, excipients, adjuvants, deodorants, diluents, preservatives, dyes/colorants, flavor enhancers, surfactants and wetting agents, dispersants, suspending agents, stabilizers, isotonizing agents, solvents, or emulsifiers that have been approved by the relevant governmental authorities for use in humans and domestic animals.

As used herein, the term "subject," "patient," "subject" or "individual" refers to an individual having a disease, disorder or condition, and the like, including mammals and non-mammals, examples of which include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment related to the methods and compositions provided herein, the mammal is a human.

The term "treatment" as used herein refers to the treatment of a disease condition associated with a mammal, particularly a human, and includes

(i) Preventing the development of a disease or condition in a mammal, particularly a mammal that has been previously exposed to the disease or condition but has not been diagnosed as having the disease or condition;

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

(iii) relieving the disease or condition, i.e., slowing the regression of the disease or condition;

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

The terms "disease" and "condition" as used herein may be used interchangeably and may have different meanings, as certain specific diseases or conditions have no known causative agent (and therefore the cause of the disease is not yet clear) and therefore are not considered as a disease but can be considered as an unwanted condition or syndrome, with more or less specific symptoms being confirmed by clinical researchers.

The terms "administering," "administration," "administering," and the like as used herein refer to methods that are capable of delivering a compound or composition to a desired site for biological action. Including, but not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

Synthesis method

The invention also provides a method for preparing the compound. The preparation of the compounds of the general formula (I) according to the invention can be carried out by the following exemplary methods and examples, which should not be construed as limiting the scope of the invention in any way. The compounds of the invention can also be synthesized using synthetic techniques known to those skilled in the art, or a combination of methods known in the art and those described herein can be used. The product of each step is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatography, and the like. The starting materials and chemical reagents required for the synthesis can be routinely synthesized or purchased according to the literature (reaxys).

Unless otherwise indicated, temperatures are in degrees celsius. Reagents were purchased from commercial suppliers such as Chemblocks Inc, Astatech Inc or mclin, and these reagents were used directly without further purification unless otherwise stated.

Unless otherwise stated, the following reactions are carried out at room temperature, in anhydrous solvents, under positive pressure of nitrogen or gas, or using a drying tube; glassware was dried and/or heat dried.

Unless otherwise stated, column chromatography purification was performed using 200-300 mesh silica gel from Qingdao oceanic chemical plants; preparation of thin-layer chromatography silica gel precast slab (HSGF254) produced by Nicoti chemical industry research institute was used; MS was measured using a Therno LCD flash model (ESI) liquid chromatography-mass spectrometer.

Nuclear magnetic data (1H NMR) Using a Bruker Avance-400MHz or Varian Oxford-400Hz nuclear magnetic spectrometer, the nuclear magnetic data was performed using CDCl as the solvent ₃ 、CD ₃ OD、D ₂ O, DMS-d6, based on tetramethylsilane (0.000ppm) or based on residual solvent (CDCl) _3: 7.26ppm；CD ₃ OD:3.31ppm；D ₂ 4.79ppm of O; d6-DMSO:2.50ppm) when indicating the diversity of the peak shapes, the following abbreviations represent the different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (doublet of doublets), dt (doublet of triplets). If the coupling constants are given, they are given in Hertz (Hz).

Example 1: preparation of (S) -7- (4-phenoxyphenyl) -5- (3-but-2-ynylaminopiperidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 1)

Step 1: synthesis of Compound 1b

The starting materials 2, 6-dichloro-3-nitro-4-aminopyridine 1a (20.8g, 0.1mol), (S) -3-Boc-aminopiperidine (22g, 0.11mol), potassium carbonate (22g, 0.2mmol) and a catalytic amount of potassium iodide and DMF (2000mL) were mixed, heated to 120 ℃ and the reaction stirred for 4 hours. Cooled to room temperature and evaporated under reduced pressure to give 1c (20.8g, 56%) as a yellow solid, LC/MS (ESI) 373M/z [ M + H ]] ⁺ 。

Step 2: synthesis of Compound 1c

After mixing the product 1b (18.59g, 0.05mol) obtained in the previous step, 4-phenoxyphenylboronic acid (10.7g, 0.05mol), tris (dibenzylideneacetone) dipalladium (4g, 4.4mmol), cesium carbonate, 1, 4-dioxane (500mL) and water (100mL), the mixture was heated to 120 ℃ under reflux, and the reaction was stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a pale yellow precipitate. The reaction mixture was diluted with water (10mL) and the solid was collected by filtration. The crude product was slurried with methanol (50mL) to give 1b as a beige solid (12.9g, 51%) which was subjected to the next reaction without further purification by LC/ms (esi) with M/z 506[ M + H ]] ⁺ 。

And step 3: synthesis of Compound 1d

N-bromosuccinimide (5.2g, 29.6mmol) was added to a solution of 1c (12.1g, 24mmol) in acetic acid (100 mL). After stirring at 60 ℃ for 2 hours, acetic acid was removed under reduced pressure. The residue was suspended in water (60mL) and saturated sodium bicarbonate solution (40mL) was added. The solid was filtered and stirred in 80 ℃ water (200mL) for 30 minutes. After cooling to ambient temperature, the solid was filtered and dried under vacuum to give crude tan solid 1d (12.2g, 87%), which was taken to the next reaction without further purification, LC/ms (esi) M/z 585[ M + H ═] ⁺ 。

And 4, step 4: synthesis of Compound 1e

Under the protection of nitrogen, 1d (7.8g, 13.35mmol), Zn (CN) ₂ (940mg, 8mmol), tris (dibenzylideneacetone) dipalladium (0.61g, 0.65mmol) and 1,1' -bis (diphenylphosphino) ferrocene (0.74g, 1.35mmol) were added to DMF/H2O (99:1, 50mL), stirred for 30 minutes, then heated to 120 ℃ and the reaction stirred for 24 hours. The resulting mixture was cooled to room temperature, and saturated NH ₄ Cl solution, concentrated ammonia water and H ₂ O (4:1:4, 10mL) precipitated. The reaction was cooled to 0 ℃ and filtered. Using saturated NH for filter cake ₄ Cl solution, concentrated ammonia water and H ₂ O (4:1:4, 2mL) and dried under vacuum to give a dark brown solid (5.38g, 76%), which was used without further purification in the next reaction, LC/MS (ESI) with M/z 531[ M + H ]] ⁺ 。

And 5: synthesis of Compound 1f

1e (5.31g, 10mmol) was dissolved in 50mL ethanol and hydrogenated over Raney nickel catalyst (2.0g) at room temperature under an atmosphere of 1.0atm H2 for 4H. After completion of the reaction, 1.6g of diatomaceous earth was added to the solution, and the mixture was stirred vigorously and filtered over a pad of diatomaceous earth. The filtrate was purified by silica gel column chromatography to give 1f (4.70g, 94%) as a dark brown solid, which was subjected to the next reaction without further purification by LC/MS (ESI) where M/z is 501[ M + H ]] ⁺ 。

Step 6: synthesis of Compound 1g

To a solution of 1f (2.5g, 5mmol) in 20ml of methanol was added sodium methoxide (0.63 g), and the mixture was stirred at room temperature for 30 minutes. A solution of diethyl oxalate (0.76g,5.1mmol) in 8ml of methanol was added dropwise to the mixture for 30 minutes, and the resulting mixture was heated to reflux for 7 h. The mixture was concentrated under reduced pressure, diluted with 30mL of water and then cooled in an ice bath. The reaction mixture was adjusted to pH 6.5 with 10% hydrochloric acid. The precipitated solid was collected by filtration, washed with water and dried to give 1g (2.6g, 94%) of the compound as a yellow solid, which was subjected to the next reaction without further purification by LC/ms (esi) with M/z 555[ M + H ]] ⁺ 。

And 7: synthesis of Compound 1h

1g (1.66g, 3mmol) of the compound of the previous step was added in portions to 80% sulfuric acid (11mL), and the reaction was stirred at 60 ℃ for 2.5 hours. After cooling to room temperature, the reaction mixture was added to ice and heated to room temperature with stirring. Adjusting the pH value to 8 with KOH, addingExtract with ethyl acetate (2 ×). Drying over anhydrous sodium sulfate, concentrating under reduced pressure to give intermediate 1H (1.40g, 97%) as a brown solid, which was then subjected to the next reaction without further purification by LC/MS (ESI) where M/z is 473[ M + H ]] ⁺ 。

And 8: synthesis of Compound 1

To a 25mL three-necked flask was added intermediate 1h (237mg, 0.5mmol) in N, N-dimethylformamide (8mL) and-2-butynoic acid (46.2mg,0.55mmol), HATU (379mg, 164mmol) and N-ethyldiisopropylamine (275 μ L) (temperature raised to 35 ℃). The final solution was to stir at room temperature for 2 hours. The mixture was diluted with ethyl acetate (10ml) and washed with water (5 ml). The organic phases were separated and the aqueous layer was extracted with ethyl acetate (2X 10 ml). The combined organic extracts were washed with washing water (containing a small amount of sodium chloride) (4X 10ml), washed with brine (10ml) and dried over anhydrous sodium sulfate. The crude product was obtained collectively under reduced pressure and purified by column chromatography to give compound 1(140mg, yield 52%) as a yellow solid. ¹ HNMR(400MHz,CD3OD)δ:7.41-7.04(m,11H),3.98-3.83(m,1H),3.19-3.07(m,1H),3.06-2.89(m,1H),3.06-2.91(m,1H),2.80(br s,1H),1.96(s,3H),1.91-1.80(m,1H),1.73(s,2H),and 1.35(s,1H)；LC/MS(ESI):m/z＝539.2[M+H] ⁺ .

Example 2: preparation of (R) -7- (4-phenoxyphenyl) -5- (3-but-2-ynylaminopiperidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 2)

Using a method similar to example 1 (intermediate was changed to (R) -3-Boc-aminopiperidine), compound 2(129mg, yield 48%, which is the final yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.41-7.04(m,9H),3.98-3.84(m,1H),3.19-3.06(m,1H),3.06-2.89(m,1H),3.08-2.90(m,1H),2.81(br s,1H),1.97(s,3H),1.91-1.81(m,1H),1.73(s,2H),and 1.36(s,1H)；LC/MS(ESI):m/z＝539.2[M+H] ⁺ .

example 3: preparation of (S) -7- (4-phenoxyphenyl) -5- (3-acrylamidopiperidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide Compound 3)

Using a method similar to example 1 (intermediate was changed to acrylic acid), Compound 3(100mg, yield 38%, this is the last step yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.43-7.04(m,9H),6.27-6.17(m,1H),6.13-6.03(m,1H),5.59(dd,1H),4.01(br s,1H),3.39(br s,1H),3.17(br s,1H),2.64(br s,1H),2.43(br s,1H),1.99-1.68(m,3H),1.35(s,1H)；LC/MS(ESI):m/z＝527.2[M+H] ⁺ .

example 4: preparation of (R) -7- (4-phenoxyphenyl) -5- (3-acrylamidopiperidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide Compound 4)

Using a method similar to example 1 (intermediate was changed to acrylic acid), Compound 4(84mg, yield 32%, this is the last step yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.43-7.04(m,9H),6.27-6.17(m,1H),6.13-6.03(m,1H),5.59(dd,1H),4.01(br s,1H),3.39(br s,1H),3.17(br s,1H),2.64(br s,1H),2.43(br s,1H),1.99-1.68(m,3H),1.35(s,1H)；LC/MS(ESI):m/z＝511.2[M+H] ⁺ .

example 5: preparation of (S) -7- (4-phenoxyphenyl) -5- (3-but-2-ynylamidopyrrolidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 5)

Using a method similar to example 1 (intermediate was changed to (S) -3-Boc-aminopyrrolidine), compound 5(147mg, yield 56%, this is the final yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.42-7.06(m,9H),4.34(m,1H),3.26-3.13(m,2H),3.11-2.97(m,2H),2.24(m,1H),1.94(s,3H),1.85(m,1H)；LC/MS(ESI):m/z＝525.2[M+H] ⁺ .

example 6: preparation of (R) -7- (4-phenoxyphenyl) -5- (3-but-2-ynylaminopyrrolidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 6)

Using a method similar to example 2 (intermediate was changed to (R) -3-Boc-aminopyrrolidine), Compound 6(160mg, 61% yield, which is the final step, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.42-7.06(m,9H),4.34(m,1H),3.26-3.13(m,2H),3.11-2.97(m,2H),2.24(m,1H),1.94(s,3H),1.85(m,1H)；LC/MS(ESI):m/z＝525.2[M+H] ⁺ .

example 7: preparation of (S) -7- (4-phenoxyphenyl) -5- (3-acrylamidopyrrolidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 7)

Using a method similar to example 2 (intermediate was changed to (S) -3-Boc-aminopyrrolidine), compound 7(97mg, yield 38%, this is the final yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.42-7.06(m,9H),6.71-6.55(d,1H),6.32-6.22(m,1H),5.74(m,1H),4.40-4.23(m,2H),3.91(dd,1H),3.86-3.58(m,3H),2.24-2.02(s,2H)；LC/MS(ESI):m/z＝513.2[M+H] ⁺ .

example 8: preparation of (R) -7- (4-phenoxyphenyl) -5- (3-acrylamidopyrrolidin-1-yl) -2, 3-dioxo-pyridine [3,4-b ] ] bipyrazinyl-8-carboxamide (Compound 8)

Using a method similar to example 1 (intermediate was changed to (R) -3-Boc-aminopyrrolidine), compound 8(92mg, yield 36%, this is the final yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.42-7.06(m,9H),6.71-6.55(d,1H),6.32-6.22(m,1H),5.74(m,1H),4.40-4.23(m,2H),3.91(dd,1H),3.86-3.58(m,3H),2.24-2.02(s,2H)；LC/MS(ESI):m/z＝513.2[M+H] ⁺ .

example 9: preparation of (S) -6- (4-phenoxyphenyl) -4- (3-but-2-alkynylaminopiperidin-1-yl) -imidazo [4,5-c ] pyridine-7-carboxamide (Compound 9)

Using a method similar to example 1 (intermediate was changed to triethyl orthoformate), compound 9(123mg, yield 52%, which is the final step, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:8.13(s,1H),7.41-7.06(m,9H),3.98-3.83(m,1H),3.19-3.07(m,1H),3.06-2.89(m,1H),3.06-2.91(m,1H),2.80(br s,1H),1.96(s,3H),1.91-1.80(m,1H),1.73(s,2H),and 1.35(s,1H)；LC/MS(ESI):m/z＝495.2[M+H] ⁺ .

example 10: preparation of (S) -6- (4-phenoxyphenyl) -4- (3-but-2-alkynylaminopiperidin-1-yl) -2-methylimidazo [4,5-c ] bipyridine-7-carboxamide (Compound 10)

Using a method similar to example 1 (intermediate was changed to acetic anhydride), Compound 9(124mg, yield 49%, which is the final step yield, the same applies hereinafter) was obtained as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.43-7.08(m,9H),3.98-3.83(m,1H),3.19-3.07(m,1H),3.06-2.89(m,1H),3.06-2.91(m,1H),2.80(br s,1H),2.54(s,3H),1.96(s,3H),1.91-1.80(m,1H),1.73(s,2H),and 1.35(s,1H)；LC/MS(ESI):m/z＝509.2[M+H] ⁺ .

example 11: preparation of 6- (4-phenoxyphenyl) -5- (1-acrylamidopiperidin-4-yl) -2-oxo-imidazo [4,5-c ] pyridine-8-carboxamide (Compound 11)

Preparation of key intermediate 4, 6-dichloro-2-oxo-imidazo [4,5-c ] pyridine-7-carboxamide 11a

N-Bromosuccinimide (5.2g, 29.6mmol) was added to 4, 6-dichloro-2-oxo-imidazole [4,5-c ]]]And pyridine (5.6g, 27mmol) in acetic acid (100 mL). After stirring at 60 ℃ for 2 hours, acetic acid was removed under reduced pressure. The residue was suspended in water (60mL) and saturated sodium bicarbonate solution (40mL) was added. The solid was filtered and stirred in 80 ℃ water (200mL) for 30 minutes. After cooling to ambient temperature, the solid was filtered and dried under vacuum to give crude 4, 6-dichloro-6-bromo-2-oxo-imidazo [4,5-c]]And 7.11g of pyridine, the yield is 93%. LC/MS (ESI) M/z 283[ M + H ]] ⁺ .

Addition of Ν, Ν, Ν ', Ν' -tetramethylethylenediamine (9g, 50.1mmol) to the product of the previous step 4, 6-dichloro-6-bromo-2-oxo-imidazo [4, 5-c)]]The bipyridine (5.64g, 20mmol) was in solution in anhydrous tetrahydrofuran (100mL) and the solution was stirred at-60 ℃ under nitrogen for 1 min. N-butyllithium (20.4mL, 50.1mmol, 2.5M in hexanes) was added slowly and the mixture stirred for 2 hours. Dry carbon dioxide gas was bubbled into the solution and the mixture was stirred at-60 ℃ for 1 hour. After heating to ambient temperature, water (100mL) was added. The tetrahydrofuran was removed under reduced pressure and the residue partitioned between ethyl acetate and water. The aqueous layer was acidified to pH 1 with 1M hydrochloric acid and the solid was filtered to give crude 4, 6-dichloro-2-oxo-imidazo [4,5-c]]3.77g of pyridine-7-carboxylic acid, yield 76%. LC/MS (ESI) M/z 249[ M + H ]] ⁺ .

4, 6-dichloro-2-oxo-imidazole [4,5-c ]]]The pyridine-7-carboxylic acid (2.49g, 10mmol) was dissolved in thionyl chloride (40ml), andthe mixture was stirred at 75 ℃ for 2 hours. Excess thionyl chloride was removed in vacuo and the residue was dissolved in anhydrous tetrahydrofuran (40 mL). Aqueous ammonia (6.0mL) was added at 0 ℃ and the mixture was stirred at ambient temperature for 10 h. The solid was filtered and recrystallized from ethanol (20mL) to give 4, 6-dichloro-2-oxo-imidazo |4,5-c]Pyridine-7-carboxamide 2.08g, yield 84%. LC/MS (ESI) M/z 248[ M + H ]] ⁺ .

Step 1: synthesis of Compound 6- (4-phenoxyphenyl-2-oxo-imidazo [4,5-c ] pyridine-8-carboxamide 11b

The raw material 4, 6-dichloro-2-oxo-imidazole [4,5-c ]]]The bipyridine-7-carboxamide 11a (3.7g, 15mmol), 4-phenoxyphenylboronic acid (6.42g, 30mmol), and tripotassium phosphate monohydrate (10.35g, 45mmol) were dissolved in dioxane (200mL) and water (20 mL). After multiple nitrogen purges, tetrakis (triphenylphosphine) palladium (2.31g, 2mmol) was added. The mixture was sparged with nitrogen for an additional 5 minutes and then heated at reflux for 24 hours. The reaction was cooled to room temperature and stirred overnight to give a pale yellow precipitate. The reaction mixture was diluted with water (10mL) and the solid was collected by filtration. The crude product was slurried with methanol (150mL) to give 1b as a beige solid (2.91g, 52%) which was subjected to the next reaction without further purification by LC/ms (esi) with M/z 382[ M + H ]] ⁺ 。

Step 2: synthesis of Compound 11c

The intermediate 6- (4-phenoxyphenyl-2-oxo-imidazole [4,5-c ] in the last step]Pyridine-8-carboxamide 11b (1.9g, 5mmol), N-Boc-1,2,5, 6-tetrahydropyridine-4-boronic acid pinacol ester (2.31g, 7.5mmol), potassium carbonate (2.07g, 15mmol), tetrakis (triphenylphosphine) palladium, 1, 4-dioxane (100mL) and water (25mL) were mixed, heated under reflux under nitrogen, and the reaction stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight, the reaction was evaporated under reduced pressure and purified by column chromatography to give beige solid 1b (1.0g, 38%) which was then reacted without further purification by LC/ms (esi) with M/z 528[ M + H ]] ⁺ 。

And step 3: synthesis of Compound 11d

To a solution of compound 11C (528mg, 1mmol) in ethyl acetate (10mL) and methanol (10mL) in the previous step was added 10% Pd/C (0.1g), and the mixture was cooled with hydrogenThe reaction was degassed 6 times and then stirred at room temperature under hydrogen atmosphere for 12 h. The solution was filtered and the filtrate was evaporated to the crude product 11d (507mg, 96%) as a brown solid, which was reacted without further purification by LC/MS (ESI) with M/z 530[ M + H ]] ⁺ 。

And 4, step 4: synthesis of Compound 11e

A reaction flask was charged with intermediate 11d (0.265g,0.5mmol) from the previous step, 2ml of ethyl acetate, 4ml of 1, 4-dioxane solution of 1N HCl. After stirring at room temperature for 2 hours, the reaction mixture was neutralized with 1N sodium hydroxide solution and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and the organic phase was evaporated to dryness under reduced pressure. Compound 11e (0.170g, 79% yield) is obtained as such in the next step, LC/MS (ESI) M/z 430.2[ M + H ]] ⁺ 。

And 5: synthesis of Compound 11

Compound 11e (129mg,0.3mmol), triethylamine (51mg,0.5mmol), and 4ml of tetrahydrofuran were added to a reaction flask, and after cooling in an ice-water bath, a solution of but-2-ynoyl chloride (45mg,0.5mmol) in 0.5ml of tetrahydrofuran was slowly added dropwise. After the addition was complete, stirring was continued for 4 hours. The reaction solution was quenched with methanol and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 11(52mg, yield 36%) as a yellow solid. ¹ H NMR(400MHz,CD ₃ OD)δ:7.43-7.08(m,9H),3.98-3.83(m,1H),3.19-3.07(m,1H),3.06-2.89(m,1H),3.06-2.91(m,1H),2.80(br s,1H),2.54(s,3H),1.96(s,3H),1.91-1.80(m,1H),1.73(s,2H),and 1.35(s,1H)；LC/MS(ESI):m/z＝484.2[M+H] ⁺ .

Examples 12 and 13: preparation of (S) -6- (4-phenoxyphenyl) -5- (3-acrylamidopiperidin-4-yl) -2-oxo-imidazo [4,5-c ] pyridine-8-carboxamide (Compound 12) and (R) -6- (4-phenoxyphenyl) -5- (3-acrylamidopiperidin-4-yl) -imidazo [4,5-c ] pyridine-8-carboxamide (Compound 13)

In a similar manner to example 11 (intermediate is replaced by 1-tert-butoxycarbonyl-3, 6-dihydro-2H-pyridine)-5-boronic acid pinacol ester) to obtain compound 9(124mg, yield 49%, which is the final yield, the same applies hereinafter) as a pale yellow solid, ¹ H NMR(400MHz,CD3OD)δ:7.43-7.08(m,9H),3.98-3.83(m,1H),3.19-3.07(m,1H),3.06-2.89(m,1H),3.06-2.91(m,1H),2.80(br s,1H),2.54(s,3H),1.96(s,3H),1.91-1.80(m,1H),1.73(s,2H),and 1.35(s,1H)；LC/MS(ESI):m/z＝509.2[M+H] ⁺ .

example 14: preparation of (S) -6- (4-phenoxyphenyl) -4- (3-but-2-alkynylamidopiperidin-1-yl) -pyrrolo [3,2-c ] pyridine-7-carboxamide (Compound 14)

Preparation of key intermediate 4, 6-dichloro-pyrrolo [3,2-c ] pyridine-7-carboxamide 14a

N-butyllithium (27.8mL, 2.5M solution in hexanes, 69.6mmol) was added to a solution of diisopropylamine (7.5g, 74.3mmol) in tetrahydrofuran (50mL) at-78 deg.C, the mixture was stirred at-78 deg.C for 30 minutes, and a solution of 2, 6-dibromo-nitropyridine (19.0g, 67.6mmol) in tetrahydrofuran (50mL) was added over 40 minutes. The mixture was stirred at-78 ℃ for 3 hours. Dry carbon dioxide was bubbled into the reaction mixture, and the mixture was stirred at ambient temperature overnight. The solvent was removed under reduced pressure, and the residue was dissolved in a mixture of ethyl acetate (50mL) and 10% aqueous sodium hydroxide (100 mL). The aqueous phase was made acidic with concentrated hydrochloric acid and extracted with ethyl acetate (3 × 150 mL). The organic layer was dried over sodium sulfate, filtered and concentrated to give intermediate 2, 6-dibromo-4-nitronicotinic acid () without further purification. LC/MS (ESI) 326[ M + H ═ M/z] ⁺ 。

2, 6-dibromo-4-nitronicotinic acid (49g, 154mmol in dry THF (1000mL), cooled to between-40 ℃ and-50 ℃ with stirring about 5mm, then vinyl magnesium bromide (in THF, 692 mL) was added dropwise692 mmol). The mixture was stirred between about-40 ℃ and-50 ℃ for about 4 h. With saturated NH ₄ Aqueous C1 (20mL) quenched the reaction. The solvent was removed under reduced pressure to obtain a residue, which was purified by preparative HPLC to give 4, 6-dibromo-1H-pyrrolo [3,2-c ]]Pyridine-7-carboxylic acid (5.4g), yield 11%. LC/MS (ESI) 321[ M + H ]] ⁺ 。

To 4, 6-dibromo-1H-pyrrolo [3,2-c]To a solution of pyridine-7-carboxylic acid (3.2g, 1.mmol) in DMF (50mL) was added HOBt (2.29g, 15mmol) and EDCl (2.88g, 15 mmol). After the reaction mixture was stirred at room temperature for about 1 hour, NH was added ₃ THF (200mL) and the resulting mixture was stirred at room temperature overnight. The suspension was then filtered and the filtrate was concentrated under reduced pressure. Water was added and extracted with ethyl acetate. The combined organic phases were washed with brine, over Na ₂ SO ₄ Dried, filtered and concentrated under reduced pressure to provide 4, 6-dibromo-1H-pyrrolo [3,2-c]Pyridine-7-carboxamide (1.53g, 48%). LC/MS (ESI) 320[ M + H ]] ⁺ 。

Step 1: synthesis of Compound 6- (4-phenoxyphenyl) -4-bromo-pyrrolo [3,2-c ] ] bipyridinyl-7-carboxamide 14b

The raw material 4, 6-dibromo-1H-pyrrolo [3,2-c]Pyridine-7-carboxamide 11a (0.64g, 2mmol), 4-phenoxyphenylboronic acid (0.86g, 4mmol) and tripotassium phosphate monohydrate (1.38g, 6mmol) were dissolved in dioxane (15mL) and water (8 mL). After multiple nitrogen purges, tetrakis (triphenylphosphine) palladium (0.35g, 0.3mmol) was added. The mixture was sparged with nitrogen for an additional 5 minutes and then heated at reflux for 24 hours. The reaction was cooled to room temperature and stirred overnight to give a pale yellow precipitate. The reaction mixture was diluted with water (10mL) and the solid was collected by filtration. The crude product was slurried with methanol (50mL) to give 14b as a beige solid (0.457g, 56%) which was subjected to the next reaction without further purification by LC/ms (esi) with M/z 409[ M + H ]] ⁺ 。

Step 2: synthesis of Compound 14c

The raw material 6- (4-phenoxyphenyl) -4-bromo-pyrrole [3,2-c]]Pyridine-7-carboxamide 14b (0.409g, 1mmol), (S) -3-Boc-aminopiperidine (0.22g, 1.1mmol), potassium carbonate (0.22g, 2mmol) and a catalytic amount of potassium iodide in DMF (20mL) were mixed, heated to 120 deg.C and the reaction 4 stirredAnd (4) hours. Cooled to room temperature and evaporated under reduced pressure to give 14c (0.27g, 51%) as a yellow solid, LC/MS (ESI) M/z 529[ M + H] ⁺ 。

And step 3: synthesis of Compound 14d

A reaction flask was charged with intermediate 14c from the previous step (0.27g,0.5mmol), 2ml of ethyl acetate, 4ml of 1, 4-dioxane solution of 1N HCl. After stirring at room temperature for 2 hours, the reaction mixture was neutralized with 1N sodium hydroxide solution and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and the organic phase was evaporated to dryness under reduced pressure. Compound 14d (0.187g, 87% yield) was obtained and used directly in the next step, LC/MS (ESI) M/z 428.2[ M + H ]] ⁺ 。

And 4, step 4: synthesis of Compound 14

Compound 14d (128mg,0.3mmol), triethylamine (51mg,0.5mmol), and 4ml of tetrahydrofuran were added to a reaction flask, and after cooling in an ice-water bath, a solution of but-2-ynoyl chloride (45mg,0.5mmol) in 0.5ml of tetrahydrofuran was slowly added dropwise. After the addition was complete, stirring was continued for 4 hours. The reaction solution was quenched with methanol and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 11(53mg, yield 36%) as a yellow solid. LC/MS (ESI) M/z 494.2[ M + H ]] ⁺ 。

Example 15: preparation of (R) -6- (4-phenoxyphenyl) -4- (3-but-2-alkynylaminopiperidin-1-yl) -pyrrolo [3,2-c ] pyridine-7-carboxamide (Compound 15)

Using a method similar to example 14 (intermediate exchanged for acetic anhydride) gave compound 15(68mg, 46% yield, this is the last step, the same applies hereinafter) as a pale yellow solid, LC/MS (ESI) 494.2[ M + H ], [ M/z ═] ⁺ .

Example 16: preparation of 6- (4-Phenoxyphenyl) -4- (1-acryloylpiperidin-4-yl) -pyrrolo [3,2-c ] pyridine-7-carboxamide (Compound 16)

Using a method similar to example 11 (intermediate exchanged for acetic anhydride) gave compound 16(59mg, 42% yield, this is the last step, the same applies hereinafter) as a pale yellow solid, LC/MS (ESI) M/z 467.2[ M + H ═ 467.2] ⁺ .

Examples 17 and 18: preparation of (S) -6- (4-phenoxyphenyl) -4- (1-acryloylpiperidin-3-yl) -pyrrolo-pyridine-7-carboxamide (Compound 17) and (R) -6- (4-phenoxyphenyl) -4- (1-acryloylpiperidin-3-yl) -pyrrolo [3,2-c ] ] pyrrolo-pyridine-7-carboxamide (Compound 18)

Using a method similar to example 16 (intermediate was changed to 1-tert-butoxycarbonyl-3, 6-dihydro-2H-pyridine-5-boronic acid pinacol ester) to obtain the racemic compound (R, S) -6- (4-phenoxyphenyl) -4- (1-acryloylpiperidin-3-yl) -pyrrole [3, 2-c)]]And pyridine-7-carboxamide 18(67mg, 49% yield, final yield, same below) as a pale yellow solid, LC/MS (ESI) with M/z 467.2[ M + H ]] ⁺ .

Example 19: in vitro activity inhibition assay for kinases BTK, BTK (R28H)

1.1 BTK inhibitory Activity screening

Using kinase buffer (50mM HEPES, 10mM MgCl) ₂ 2mM DTT, 1mM EGTA, 0.01% Tween20), 350ng/uL of BTK mother solution is diluted, 6 uL of 1.67 X0.134 ng/uL of working solution (final concentration is 0.08 ng/uL) is added into each well, different compounds 101-128 dissolved in DMSO are added into the wells by a nanoliter loading instrument, the final concentration of the compounds is 1000nM-0.244nM, the final concentration of positive drugs is 50nM-0.0122nM, 4-fold gradient is carried out, 7 concentrations are obtained, meanwhile, a blank control well (containing no enzyme) and a negative control well (containing enzyme and adding DMSO as a solvent) are arranged, and 2 duplicate wells are arranged. After the enzyme reacts with the compound or the solvent for 30min, 5 X250. mu. MATP (final concentration of 50uM) prepared by using a kinase buffer solution and 5 X0.5. mu.M substrate (final concentration of 0.1. mu.M, ULight-poly GT) are mixed according to a ratio of 1:1, and 4. mu.L of the substrate per well is added into the well; after the plate is sealed by a sealing film, the reaction is carried out for 2 hours at room temperatureAfter incubation, 5 μ L of 4 × 8nM detection reagent (2 nM final concentration, Ab) was added to each well and incubated for 1 hour at room temperature; the plate was read with a PE instrument (excitation 620nm, emission 665 nm). Calculating the inhibition ratio, and calculating IC ₅₀ The value is obtained. The results of the measurements are shown in the following table

1.2 BTK (R28H) inhibitory Activity screening

200ng/uL of BTK (R28H) stock solution was diluted with kinase buffer (50mM HEPES, 10mM MgCl2, 2mM DTT, 1mM EGTA, 0.01% Tween20), 6. mu.L of 1.67 X1.67 ng/uL working solution (final concentration of 1 ng/uL) was added to each well, and various compounds 101-128 dissolved in DMSO were added to the wells using a nanoliter loader so that the final concentration of the compounds was 2000nM-0.488nM and the final concentration of the positive drug was 200nM-0.0488nM, with a 4-fold gradient for a total of 7 concentrations, while blank control wells (containing no enzyme) and negative control wells (containing enzyme, vehicle DMSO) were set for 2 replicate wells. After the enzyme reacts with the compound or the solvent for 30min, 5 X500. mu. MATP (final concentration of 100uM) prepared by using a kinase buffer solution and 5 X0.5. mu.M substrate (final concentration of 0.1. mu.M, ULight-poly GT) are mixed according to a ratio of 1:1, and 4. mu.L of the substrate per well is added into the well; after the plate was sealed with a membrane, after reacting at room temperature for 2 hours, 5. mu.L of 4X 40mM EDTA (final concentration: 10mM) was added to each well for 5min at room temperature, and 5. mu.L of 4X 8nM detection reagent (final concentration: 2nM, Ab) was further added to each well and incubated at room temperature for 1 hour; the plate was read with a PE instrument (excitation 620nm, emission 665 nm). Calculating the inhibition ratio, and calculating IC ₅₀ The value is obtained. The results of the assay are shown in the following table, which shows the activity data of compounds 1-28 for wild-type BTK, mutant BTK (R28H). Active utilization of IC ₅₀ Characterization, wherein "A" represents IC ₅₀ Less than or equal to 10 nM; "B" means 10<IC ₅₀ Less than or equal to 100 nM; "C" means 100<IC ₅₀ Less than or equal to 500 nM; "D" means 500<IC ₅₀ ≤2000nM。

Claims (10)

1. A compound having the general formula (I) or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, polymorph or isomer thereof,

X ₁ ，X ₂ ，X ₄ may be independently selected from N, CR ¹ ；

X ₃ May be independently selected from absent N, CR ¹ ；

The bonds a and b are single bonds or double bonds;

Ar ₁ and Ar ₂ Independently selected from a phenyl ring or a 5-6 membered heteroaromatic ring wherein said phenyl and heteroaromatic rings are optionally substituted by one or more G ¹ Substituted;

R ¹ independently selected from H, cyano, halogen, C _1-6 Alkyl, COOH, CONH2, NHCOH, CONHR2, OR ² or-NHR ² ；

R ² Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -OR ³ 、-NR ³ R ⁴ 、-C(O)NR ³ R ⁴ Wherein said alkyl, cycloalkyl OR heterocycloalkyl is optionally substituted by cyano, halogen, -OR ⁵ 、-NR ⁵ R ⁶ 、C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocycloalkyl;

u and W are independently selected from-C _0-4 Alkyl-, -CR ⁷ R ⁸ -、-C _1-2 Alkyl (R) ⁷ )(OH)-、-C(O)-、-CR ⁷ R ⁸ O-、-OCR ⁷ R ⁸ -、-SCR ⁷ R ⁸ -、-CR ⁷ R ⁸ S-、-NR ⁷ -、-NR ⁷ C(O)-、-C(O)NR ⁷ -、-NR ⁷ C(O)NR ⁸ -、-CF ₂ -、-O-、-S-、-S(O) _m -、-NR ⁷ S(O) ₂ -、-S(O) ₂ NR ⁷ -；

Y is absent or C is selected _3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spiro cyclic group, 5-12 membered spiro heterocyclic group, aromatic group or heteroaromatic group, wherein said cycloalkyl, heterocycloalkyl, spiro cyclic group, fused heterocyclic group, spiro heterocyclic group, aromatic group or heteroaromatic group is optionally substituted with one or more G ¹ Substituted;

z is independently selected from cyano, -NR ¹² CN、

Bond c is a double or triple bond;

when c is a double bond, R ^a 、R ^b And R ^c Each independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl. Wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ² Substituted;

R ^a and R ^b Or R ^b And R ^c Optionally taken together with the carbon atom to which they are attached to form a 3-6 membered ring optionally containing heteroatoms;

when the bond c is a triple bond, R ^a And R ^c Is absent, R ^b Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl by one or more G ³ Substituted;

R ¹² independently selected from H, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl, wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ⁴ Substituted;

G ¹ 、G ² 、G ³ and G ⁴ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹³ 、-OC(O)NR ¹³ R ¹⁴ 、-C(O)OR ¹³ 、-C(O)NR ¹³ R ¹⁴ 、-C(O)R ¹³ 、-NR ¹³ R ¹⁴ 、-NR ¹³ C(O)R ¹⁴ 、-NR ¹³ C(O)NR ¹⁴ R ¹⁵ 、-S(O) _m R ¹³ or-NR ¹³ S(O) _m R ¹⁴ Wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by 1 or more cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹⁶ 、-OC(O)NR ¹⁶ R ¹⁷ 、-C(O)OR ¹⁶ 、-C(O)NR ¹⁶ R ¹⁷ 、-C(O)R ¹⁶ 、-NR ¹⁶ R ¹⁷ 、-NR ¹⁶ C(O)R ¹⁷ 、-NR ¹⁶ C(O)NR ¹⁷ R ¹⁸ 、-S(O) _m R ¹⁶ or-NR ¹⁶ S(O) _i R ¹⁷ Substituted with the substituent(s);

R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ 、R ⁹ 、R ¹⁰ 、R ¹¹ 、R ¹³ 、R ¹⁴ 、R ¹⁵ 、R ¹⁶ 、R ¹⁷ and R ¹⁸ Each independently selected from hydrogen, cyano, halogen, C _1-6 Alkyl radical, C _3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl; and is

m is 1 or 2.

2. A compound according to claim 1 or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, polymorph or isomer thereof and a mixture form thereof.

3. Which is selected from the group consisting of the following compounds,

or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, isomer, and mixtures and forms thereof.

4. A pharmaceutical composition comprising a novel Bruton's tyrosine kinase inhibitor of claim 1 or 3, or an isomer, hydrate, stable isotope derivative, solvate, polymorph, pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is in the form of a tablet, capsule, granule, spray or injection.

6. The pharmaceutical composition of claim 4, wherein the pharmaceutically acceptable carrier is selected from one or more of a filler, a disintegrant, a binder, and a lubricant.

7. Use of a novel Bruton's tyrosine kinase inhibitor according to claims 1 and 3, or an isomer, hydrate, solvate, polymorph, pharmaceutically acceptable salt thereof, as a protein tyrosine kinase inhibitor.

8. Use according to claim 7, characterized in that: the protein tyrosine kinase inhibitor is Bruton's tyrosine kinase inhibitor.

9. Use of a novel Bruton's tyrosine kinase inhibitor of claims 1 and 3, or an isomer, hydrate, solvate, polymorph, pharmaceutically acceptable salt thereof, or a pharmaceutical composition of any of claims 4-6 for the treatment or prevention of a Bruton's tyrosine kinase associated disease.

10. Use according to claim 9, characterized in that: the Bruton's tyrosine kinase related diseases are selected from the group consisting of: acute Lymphocytic Leukemia (ALL), Chronic Myelocytic Leukemia (CML), Mantle Cell Lymphoma (MCL), carcinoma of large intestine, rheumatoid arthritis, organ transplantation rejection, psoriasis, lupus erythematosus, etc.