WO2017162007A1 - Aromatic amide derivative and preparation method and medicinal application thereof - Google Patents

Abstract

The present invention relates to an aromatic amide derivative and a preparation method and medicinal application thereof. Specifically, the present invention relates to a novel derivative represented by general formula (I) and a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same, and a preparation method therefor. The present invention further relates to use of the derivative and the pharmaceutically acceptable salt thereof or the pharmaceutical composition comprising the same in the preparation of a therapeutic agent, particularly a Bruton's tyrosine kinase inhibitor, and in the preparation of a medicament for treating and/or preventing a disease such as a tumor or an immune-related disease. The substituents in the general formula (I) are as defined in the specification.

Description

Aromatic amide derivatives, preparation method thereof and application thereof in medicine Technical field

The present invention relates to a novel aromatic amide derivative, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, and a process for the preparation thereof. The present invention also relates to the above-mentioned derivatives and pharmaceutically acceptable salts thereof or pharmaceutical compositions containing the same, in the preparation of therapeutic agents, Bruton tyrosine kinase inhibitors, and in the preparation of diseases for the treatment and/or prevention of diseases such as tumors and inflammation Use in medicine.

Background technique

Bruton tyrosine kinase (BTK) is an important enzyme that mediates cellular signaling and is present in plasma cells, including B cells. B cells are activated by the B cell receptor (BCR), and BTK plays a decisive role in the BCR-mediated signaling pathway. Activation of BCR on B cells causes activation of BTK, leading to an increase in downstream phospholipase C (PLC) concentration and activation of the IP3 and DAG signaling pathways. This signaling pathway promotes cell proliferation, adhesion and survival and plays an important role in the development of B-cell lymphoma.

The presence of BTK mutants in the population demonstrates that BTK can be a drug target. Affinity-free agammaglobulinemia (XLA) is a rare genetic disease found in 1/250,000 men. In this disease, the function of BTK is inhibited, resulting in the production or maturation of B cells. Men with XLA disease have almost no B cells in their bodies, and there are few circulating antibodies, which are prone to serious or even fatal infections. Thus, BTK plays an extremely important role in the growth and differentiation of B cells.

By inhibiting the activity of BTK, BTK inhibitors can inhibit the proliferation of B lymphoma cells, destroy the adhesion of tumor cells, promote the apoptosis of tumor cells, and make Btk a attractive drug target in B cell-associated cancers. Especially for B-cell lymphomas and leukemias, such as non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), and anti-recurrent or refractory mantle cell lymphoma ( Mantle cell lymphoma, MCL) and so on. The only drug on the market with specific inhibition of BTK is Ibrutinib from Pharmacyclics/JNJ. Ibrutinib is a non-reversible small molecule BTK inhibitor, which has very obvious therapeutic effect on the treatment of MCL, CLL and WM, and is safe. Other BTK inhibitors that enter clinical lymphomas include Celgene's CC-292, Acerta's ACP-196, ONO's ONO-4059, and Beigene's BGB-3111.

In addition to anti-B cell lymphoma and leukemia, BTK inhibitors can also inhibit B cell autoantibodies and cytokine production. In autoimmune diseases, B cells present autoantigens, promote T cell activation and secretion, cause inflammatory factors to cause tissue damage, and activate B cells to produce large amounts of antibodies, triggering autoimmune responses. The interaction of T and B cells forms a positive feedback regulatory chain, leading to uncontrolled autoimmune response and aggravation of histopathological damage. Studies have shown that regulatory B cells exist in the body, which can negatively regulate immune responses and inhibit immune-mediated inflammatory responses by secreting interleukin-10 (IL-10) or transforming growth factor beta 1 (TGF-β1) and other mechanisms. . Therefore, B cells can be used as autoimmune diseases, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), allergic diseases (such as esophagitis, eosoniphilic esophagitis). Target. There are currently no specific BTK inhibitors for immune diseases on the market, but several are in clinical stages, such as CC-292 from Celgene, ACP-196 from Acerta, HM-71224 from Hanmi, and PRN- from Principia. 1008, a compound of Merck Serono's M-2951 and Pharmacyclics.

The global market for non-Hodgkin's lymphoma will grow from $3.3 billion in 2007 to $4.7 billion in 2017 (an average annual increase of 3.7%). As a B-cell antibody-based biological preparation,

Sales in the NHL treatment sector will grow from $2.8 billion in 2007 to $3.2 billion in 2017. But as a biological agent,

And Lymphostat B, there are deficiencies in the wide range of B cell clearance, and there is no suitable oral dosage. The potential market for autoimmune diseases such as antirheumatic oral drugs is even greater. Now based on anti-tumor necrosis factor (TNF) drugs, the market is $ 5 billion, but currently based on anti-TNF drugs, such as

with

It is specific for T cells and is administered by subcutaneous injection. The market for rheumatoid arthritis treatment will grow about twice from 2009 to 2017, reaching $13.4 billion. The potential market for other disease treatments, such as lupus erythematosus, is also very large. Therefore, BTK inhibitors have a very broad prospect.

Due to the important role played by BTK in multiple signaling pathways, the development of BTK inhibitors has attracted the attention of many biopharmaceutical companies. A series of patent applications for BTK inhibitors have been published, including WO2007087068, WO2010126960, WO2011019780, WO2011090760, WO2012135801, WO2012158764, WO2013060098, WO2013081016, WO2013010869, WO2013113097, CN103113375, WO2014068527, WO2014125410, WO2014173289, WO2013118986, WO2015017502 and WO2015048689, etc., but still need to develop new compounds with better pharmacodynamics. Through continuous efforts, the present invention has designed a compound having a structure represented by the general formula (I), and it has been found that a compound having such a structure exhibits an excellent effect and effect.

Summary of the invention

The object of the present invention is to provide a compound represented by the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer thereof, or a mixture thereof. Form, and its pharmaceutically acceptable salts:

among them:

Rings A and B ¹ are each independently selected from aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted by one or more G ¹ ;

B ^{2 is} independently selected from H, C _3-8 cyclo, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein the cyclo, heterocyclyl, aryl or heteroaryl is optionally Or replaced by multiple G ² ;

L ^{1 is} independently selected from -C _0-2 alkyl-, -CR ² R ³ -, -C _1-2 alkyl (R ¹ )(OH)-, -C(O)-, -CR ¹ R ² O-, -OCR ¹ R ² -, -SCR ¹ R ² -, -CR ¹ R ² S-, -NR ¹ -, -NR ¹ C(O)-, -C(O)NR ¹ -, -NR ¹ CONR ² -, -CF ₂ -, -O-, -S-, -S(O) _m -, -NR ¹ S(O) _m - or -S(O) _m NR ¹ -;

L ^{2 is} independently selected from -C _0-4 alkyl-, -C(O)-, -O-, -NR ³ -, -NR ³ C(O)- or -NR ³ S(O) _m -;

X is independently selected from C _0-4 alkyl, C _3-8 cyclo, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein the alkyl, cyclo, heterocyclyl, aryl or The heteroaryl group is optionally substituted with one or more G ³ ;

Y is independently selected from -C(O)-, -NR ⁴ C(O)-, -S(O) _m - or -NR ⁴ S(O) _m -;

R is independently selected from H, D, alkyl, cyclo, heterocyclyl, aryl or heteroaryl, wherein the alkyl, cyclo, heterocyclyl, aryl or heteroaryl is optionally taken by one or Replaced by multiple G ⁴ ;

R ¹ , R ² , R ³ and R ⁴ are each independently selected from H, D, C _1-8 alkyl, C _3-8 cyclic, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein The alkyl, cyclo, heterocyclyl, aryl or heteroaryl group is optionally substituted by one or more G ⁵ ;

G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from H, D, halogen, cyano, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl , -OR ⁵ , -OC(O)NR ⁵ R ⁶ , -C(O)OR ⁵ , -C(O)NR ⁵ R ⁶ , -C(O)R ⁵ , -NR ⁵ R ⁶ , -NR ⁵ C(O)R ⁶ , -NR ⁵ C(O)NR ⁶ R ⁷ , -S(O) _m R ⁵ or -NR ⁵ S(O) _m R ⁶ , wherein the alkyl, alkenyl, alkynyl group Or a cycloalkyl, heterocyclyl, aryl or heteroaryl group optionally selected from one or more selected from the group consisting of D, halogen, cyano, C _1-8 alkyl, C _3-8 cycloalkyl, 3-8-membered Ring group, -OR ⁷ , -OC(O)NR ⁷ R ⁸ , -C(O)OR ⁷ , -C(O)NR ⁷ R ⁸ , -C(O)R ⁷ , -NR ⁷ R ⁸ ,- Substituted with a substituent of NR ⁷ C(O)R ⁸ , —NR ⁷ C(O)NR ⁸ R ⁹ , —S(O) _m R ⁷ , —NR ⁷ S(O) _m R ⁸ ;

R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ are each independently selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-8 cycloalkyl, 3-8 membered monocyclic hetero a cyclic group, a monocyclic heteroaryl group or a monocyclic aryl group;

m is 1 or 2.

In one embodiment of the invention, a compound of the formula (I) or a tautomer, a mesomer, a racemate, an enantiomer or a diastereomer thereof And a mixture thereof, and a pharmaceutically acceptable salt thereof, which is a compound of the formula (II) or a tautomer thereof, a mesogen, a racemate, an enantiomer, a non-pair Isomers, mixtures thereof, and pharmaceutically acceptable salts thereof:

among them:

Ring B is

Z ¹ and Z ² are each independently selected from CH or N;

Z ³ is O or S;

The definitions of B ¹ , B ² , L ¹ , L ² , X, Y and R are as set forth in claim 1.

In another embodiment of the present invention, a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof And a mixture thereof, and a pharmaceutically acceptable salt thereof, which is a compound of the formula (III) or a tautomer thereof, a mesogen, a racemate, an enantiomer, a non- Enantiomers, mixtures thereof, and pharmaceutically acceptable salts thereof:

among them:

B ¹ and B ² are a benzene ring or a six-membered heteroaryl ring;

Rings B, L ² , X, Y, R, G ¹ and G ² are as defined in claims 1-2.

In another embodiment of the present invention, a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof And mixtures thereof, and pharmaceutically acceptable salts thereof, which are compounds of the formula (IV) or tautomers, meso, racemates, enantiomers, non- Enantiomers, mixtures thereof, and pharmaceutically acceptable salts thereof:

among them:

P ¹ and P ² are each independently selected from C(R ^a ) or N;

R ^a is H or an alkyl group;

n and o are each independently selected from 0, 1 or 2;

Rings B, B ¹ , B ² , L ² , Y and R are as defined in claims 1-3.

In another embodiment of the present invention, a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof And mixtures thereof, and pharmaceutically acceptable salts thereof, wherein R is independently selected from H, D, alkyl or heteroalkyl.

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

Or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable salt thereof.

The invention further relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer, a non- Enantiomers, mixtures thereof, and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers, diluents and excipients.

Another aspect of the invention relates to a compound of the formula (I) or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer, a mixture thereof And a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in the preparation of a Bruton tyrosine kinase inhibitor.

Another aspect of the invention relates to a compound of the formula (I) or a tautomer, a mesophil, a racemate, an enantiomer, a diastereomer, a mixture thereof And a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for use in the manufacture of a medicament for the treatment and/or prevention of diseases such as tumors and inflammation.

The present invention also relates to a method for treating and/or preventing a disease such as a tumor and inflammation, which comprises administering to a patient in need of treatment a therapeutically effective amount of a compound of the formula (I) or a tautomer thereof, internal elimination A rot, a racemate, an enantiomer, a diastereomer, a mixture thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

Another aspect of the invention relates to a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer thereof, which is a medicament for the treatment and/or prevention of diseases such as tumors and inflammation. Isomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof.

detailed description

Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

The expression "C _xy " as used herein denotes a range of the number of carbon atoms, wherein x and y are both integers, for example, a C _3-8 cyclo group represents a cyclic group having 3 to 8 carbon atoms, -C _0-2 The alkyl group means an alkyl group having 0 to 2 carbon atoms, and the -C ₀ alkyl group means a chemical single bond.

"Alkyl" means a saturated aliphatic hydrocarbon group, including straight chain and branched chain groups of 1 to 20 carbon atoms, for example It may be a linear or branched group of 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. As used herein, "alkyl" can be a monovalent, divalent or trivalent group. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2 -methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, and various branched isomers thereof. Non-limiting examples also include methylene, methine, ethylene, sec-ethyl, propylene, propylene, butylene, butyl, and various branched isomers thereof. The alkyl group can be optionally substituted or unsubstituted.

"Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising from 3 to 12 ring atoms, for example from 3 to 12, from 3 to 10 or from 3 to 6 ring atoms, Or it can be a 3, 4, 5, or 6-membered ring. Non-limiting examples of monocyclic ring groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Base, cyclooctyl and the like. The cyclic group can be optionally substituted or unsubstituted.

"Heterocyclyl" means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising from 3 to 20 ring atoms, for example 3 to 16, 3 to 12, 3 to 10, 3 Up to 6 or 5 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) _m (where m is an integer from 0 to 2), but excluding -OO-, - The ring portion of OS- or -SS-, the remaining ring atoms are carbon. Preferably, it comprises from 3 to 12 ring atoms, wherein from 1 to 4 are heteroatoms, more preferably the heterocycloalkyl ring contains from 3 to 10 ring atoms, most preferably a 5- or 6-membered ring, of which 1 to 4 are hetero More preferably, one to three atoms are hetero atoms, and most preferably one or two are hetero atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, azetidinyl and the like. Polycyclic heterocyclic groups include spiro, fused, and bridged heterocyclic groups.

"Spiroheterocyclyl" means a polycyclic heterocyclic group of 5 to 20 members in which one atom (referred to as a spiro atom) is shared between monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) _m The hetero atom (where m is an integer from 0 to 2) and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspirocyclic group according to the number of common spiro atoms between the ring and the ring, and is preferably a monospirocycloalkyl group and a bispirocycloalkyl group. More preferably, it is 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6-membered monospirocycloalkyl group. Non-limiting examples of spirocycloalkyl groups include

"Fused heterocyclyl" refers to 5 to 20 members, each ring of the system sharing an adjacent pair of atoms of a polycyclic heterocyclic group with other rings in the system, and one or more rings may contain one or more a bond, but none of the rings have a fully conjugated π-electron system in which one or more ring atoms are selected from nitrogen, oxygen or S(O) _m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. It may be classified into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl group according to the number of constituent rings, preferably a bicyclic or tricyclic ring, more preferably a 5-member/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. . Non-limiting examples of fused heterocyclic groups include

The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring wherein the ring to which the parent structure is attached is a heterocyclic group, non-limiting examples comprising:

Wait.

The heterocyclic group may be optionally substituted or unsubstituted.

"Aryl" means a 6 to 14 membered all-carbon monocyclic or fused polycyclic ring (ie, a ring that shares a pair of adjacent carbon atoms), a polycyclic ring having a conjugated π-electron system (ie, with adjacent pairs) The ring group of a carbon atom is preferably 6 to 10 members, such as a phenyl group and a naphthyl group, and most preferably a phenyl group. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring wherein the ring to which the parent structure is attached is an aryl ring, non-limiting examples comprising:

The aryl group can be substituted or unsubstituted.

"Heteroaryl" refers to a heteroaromatic system containing from 1 to 4 heteroatoms, from 5 to 14 ring atoms, wherein the heteroatoms include oxygen, sulfur and nitrogen. It is preferably 5 to 10 yuan. More preferably, the heteroaryl group is 5- or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, oxazolyl, Isoxazolyl, thiazolyl, isothiazolyl, etc., the heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring to which the parent structure is attached is a heteroaryl ring Non-limiting examples include:

The heteroaryl group can be optionally substituted or unsubstituted.

"Halogen" means fluoro, chloro, bromo or iodo.

"Cyano" means -CN.

"Optional" or "optionally" means that the subsequently described event or environment may, but need not, occur, including where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may be, but not necessarily, present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group. .

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms, independently of each other, substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

"Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers. And excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient. Biological activity.

Example

The preparation of the compound of the formula (I) of the present invention or a pharmaceutically acceptable salt thereof can be carried out by the exemplary methods described in the following examples and related publications used by those skilled in the art, but these examples It is not intended to limit the scope of the invention.

The structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR was measured using a Bruker AVANCE-400 or Varian Oxford-300 nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDC1 ₃ ), deuterated methanol (CD ₃ OD). The internal standard is tetramethylsilane (TMS) and the chemical shift is given in units of 10 ^-6 (ppm).

The MS was measured using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (manufacturer: Shimadzu, model: 2020).

The HPLC was measured using an Agilent 1200 DAD high pressure liquid chromatograph (Sunfirc C18, 150 x 4.6 mm, 5 μm, column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4.6 mm, 5 μm column).

The thin-layer chromatography silica gel plate uses Qingdao Ocean GF254 silica gel plate, and the silica gel plate used for thin-layer chromatography (TLC) has a specification of 0.15 mm to 0.2 mm. The specification for separation and purification of thin layer chromatography is 0.4 mm to 0.5 mm. silicone board.

Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as a carrier.

The known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Beijing Coupling Companies such as chemicals.

In the examples, unless otherwise specified, the reactions were all carried out under an argon atmosphere or a nitrogen atmosphere.

An argon atmosphere or a nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon having a volume of about 1 L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.

The pressurized hydrogenation reaction was carried out using a GCD-500G high purity hydrogen generator and a BLT-2000 medium pressure hydrogenator from Beijing Jiawei Kechuang Technology Co., Ltd.

The hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times.

The microwave reaction used a CEM Discover-SP type microwave reactor.

In the examples, unless otherwise specified, the temperature of the reaction is room temperature, and the temperature range is from 20 ° C to 30 ° C.

The reaction progress in the examples was monitored by thin layer chromatography (TLC), and the system used for the reaction was A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system, the volume ratio of the solvent was based on The polarity of the compound is adjusted to adjust.

The system for purifying the compound using the column chromatography eluent and the system for developing the thin layer chromatography include A: dichloromethane and methanol systems; B: petroleum ether and ethyl acetate system, the volume ratio of the solvent according to the compound The polarity is adjusted to adjust, and a small amount of triethylamine and an acidic or alkaline reagent may be added for adjustment.

resolve resolution:

The compound can be obtained according to the following classical amide coupling reaction, and the amine precursor can be obtained according to the synthesis method of the patent WO2015048662.

Example 1

6-(1-(but-2-ynyl)piperidin-4-yl)-2-(4-phenoxyphenyl)nicotamide

first step

2,6-dichloronic acid amide

The compound 2,6-dichloro nicotine nitrile 1a (1.73 g, 10 mmol), concentrated sulfuric acid (10 mL) and water (2 mL) were combined and then warmed to 90 ° C and stirred for 1 hour. After cooling to room temperature, the reaction mixture was poured into an ice water bath, and the pH was adjusted to 8 with aqueous ammonia. Filtration, the filter cake was washed with water and dried to give the desired product 2,6-dichloronic acid amide 1b (1.4 g, solid), yield: 73%.

MS m/z (ESI): 191 [M+1]

Second step

6-chloro-2-(4-phenoxyphenyl)nicotamide

Compound 2,6-dichloronic acid amide 1b (0.678 g, 3.5 mmol), (4-phenoxyphenyl)boronic acid (0.835 g, 3.9 mmol), cesium carbonate (0.977 g, 7.1 mmol), three (two) The mixture was mixed with palladium (0.324 g, 0.35 mmol), water (4 mL) and 1,4-dioxane (15 mL). The mixture was cooled to room temperature, and then evaporated to dryness. (0.611 g, solid), Yield: 53%.

MS m/z (ESI): 325 [M+1]

third step

5-carbamoyl-6-(4-phenoxyphenyl)-5',6'-dihydro-[2,4'-bipyridyl]-1'(2'H)-carboxylic acid tert-butyl ester

The compound 6-chloro-2-(4-phenoxyphenyl)nicotinamide 1c (0.162 g, 0.50 mmol), (1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridine 4-yl)boronic acid (0.232 g, 0.75 mmol), tetrakis(triphenylphosphine)palladium (0.115 g, 0.1 mmol), potassium carbonate (0.207 g, 1.5 mmol), ethylene glycol dimethyl ether (10 mL) and After mixing with water (2 mL), it was heated to 90 ° C and stirred for 5 hours. The mixture was cooled to room temperature, and then evaporated to dryness. 5',6'-Dihydro-[2,4'-bipyridyl]-1'(2'H)-carboxylic acid tert-butyl ester 1d (0.21 g, solid), yield: 89%.

MS m/z (ESI): 472 [M+1]

the fourth step

4-(5-carbamoyl-6-(4-phenoxyphenyl)pyridin-2-yl)piperidine-1-carboxylic acid tert-butyl ester

The compound 5-carbamoyl-6-(4-phenoxyphenyl)-5',6'-dihydro-[2,4'-bipyridyl]-1'(2'H)-carboxylic acid The butyl ester 1d (0.21 g, 0.45 mmol), palladium charcoal (10 mg), and ethyl acetate (10 mL) were combined and evaporated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 70/1) to afford the desired product 4-(5-carbamoyl-6-(4-phenoxyphenyl)pyridine- tert-Butyl 2-ethylpiperidine-1-carboxylate 1e (0.20 g, solid), yield: 94%.

MS m/z (ESI): 474 [M+1]

the fifth step

2-(4-phenoxyphenyl)-6-(piperidin-4-yl)nicotinamide

The compound 4-(5-carbamoyl-6-(4-phenoxyphenyl)pyridin-2-yl)piperidine-1-carboxylic acid tert-butyl ester 1e (0.215 g, 0.45 mmol), trifluoroacetic acid (2 mL) was mixed with dichloromethane (5 mL) and stirred at room temperature for 1 hour. The solution was dissolved in vacuo and the residue was crystallised eluted eluted eluted The organic phase was dried over anhydrous sodium sulfate, filtered, evaporated, evaporated, evaporated, evaporated, -Phenoxyphenyl)-6-(piperidin-4-yl)nicotinamide 1f (0.12 g, mp.

MS m/z (ESI): 374 [M+1]

Step 6

6-(1-(but-2-ynyl)piperidin-4-yl)-2-(4-phenoxyphenyl)nicotamide

The compound 2-(4-phenoxyphenyl)-6-(piperidin-4-yl)nicotinamide 1f (100 mg, 0.27 mmol), buty-2-ynoic acid (34 mg, 0.405 mmol), 2- 7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (114 mg, 0.3 mmol), triethylamine (55 mg, 0.54 mmol) and N,N- Dimethylformamide (10 mL) was mixed and stirred at room temperature for 3 hours. The residue was purified by silica gel column chromatography (dichloromethane/methanol=50/1) to give the desired product 6-(1-(but-2- </RTI> </RTI> <RTIgt; -(4-Phenoxyphenyl)nicotinamide 1 (54 mg, solid), yield: 46%.

MS m/z (ESI): 440 [M+1]

¹ H NMR (400 MHz, CD 3 OD) δ 7.87 (d, J = 7.9 Hz, 1H), 7.79 - 7.67 (m, 2H), 7.45 - 7.29 (m, 3H), 7.17 (t, J = 7.4 Hz, 1H) ), 7.12–7.00 (m, 4H), 4.59 (dd, J = 18.1, 15.6 Hz, 2H), 3.38 (d, J = 2.8 Hz, 1H), 3.13 (tt, J = 11.8, 3.6 Hz, 1H) , 2.91 (td, J = 12.9, 3.0 Hz, 1H), 2.14 - 1.97 (m, 5H), 1.82 (m, 2H).

Example 2

2-(4-phenoxyphenyl)-6-(1-propyridylpiperidin-4-yl)nicotinamide

Example 2 was synthesized by referring to the procedure of Example 1, but in the sixth step, butynoic acid was substituted with propiolic acid.

MS m/z (ESI): 426 [M+1]

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.08 (d, J = 7.9 Hz, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.40 (t, J = 7.9 Hz, 2H), 7.28 - 7.16 (m, 2H), 7.10 (d, J = 8.3 Hz, 4H), 5.54 (d, J = 61.1 Hz, 2H), 4.73 (d, J = 13.3 Hz, 1H), 4.56 (d, J = 13.7 Hz) , 1H), 3.31 (t, J = 11.7 Hz, 1H), 3.18 (d, J = 19.3 Hz, 2H), 2.86 (td, J = 13.0, 2.7 Hz, 1H), 2.11 (dd, J = 23.9, 13.5 Hz, 2H), 2.00 - 1.74 (m, 2H).

Example 3

6-(1-(but-2-ynyl)pyrrolidin-3-yl)-2-(4-phenoxyphenyl)nicotinamide

Example 3 was synthesized by referring to the procedure of Example 1, but in the third step, tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-(2-)2-dihydro-1H-pyrrole-1-carboxylate substituted (1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)boronic acid .

MS m/z (ESI): 426 [M+1]

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.05 (t, J = 7.9Hz, 1H), 7.71 (dd, J = 8.5,5.6Hz, 2H), 7.44-7.34 (m, 2H), 7.27-7.15 ( m, 2H), 7.13 - 6.99 (m, 5H), 5.77 - 5.43 (m, 2H), 4.16 (dd, J = 10.8, 7.5 Hz, 0.5H), 4.03 - 3.92 (m, 2H), 3.76 (ddd , J = 26.2, 15.4, 7.6 Hz, 3H), 3.56 (dt, J = 12.4, 8.0 Hz, 0.5H), 2.46 - 2.29 (m, 2H), 2.01 (d, J = 13.1 Hz, 3H), 1.49 (dd, J = 16.2, 9.7 Hz, 1H).

Example 4

6-(4-(but-2-ynyl)piperazin-1-yl)-2-(4-phenoxyphenyl)nicotamide

first step

4-(6-chloro-5-cyanopyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester

Compound 2,6-dichloronicotonitrile 1a (0.519 g, 3.0 mmol), piperazine-1-carboxylic acid tert-butyl ester (0.558 g, 3.0 mmol), potassium carbonate (0.636 g, 6.0 mmol) and ethanol (30 mL) The mixture was heated to 80 ° C and stirred under reflux for 3 hours. The mixture was cooled to room temperature, and then evaporated to dryness. EtOAcjjjjjjjjjj Tert-butyl piperazine-1-carboxylic acid 4a (0.59 g, solid), yield: 60%.

MS m/z (ESI): 323 [M+1]

Second step

2-chloro-6-(piperazin-1-yl) nicotinamide

4-(6-chloro-5-cyanopyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester 4a (0.60 g, 1.86 mmol) was used as a starting material, and was synthesized by the synthesis method of 1b in Example 1, The title product 2-chloro-6-(piperazin-1-yl)niconamide 4b (0.42 g, m.

MS m/z (ESI): 241 [M+1]

third step

2-(4-phenoxyphenyl)-6-(piperazin-1-yl)nic acid amide

2-chloro-6-(piperazin-1-yl)nic acid amide 4b (0.29 g, 1.2 mmol) was used as a starting material, which was synthesized by the method of 1c of Example 1 to give the title product 2-(4-phenoxy). Phenyl)-6-(piperazin-1-yl)nicotinamide 4c (0.26 g, solid), yield: 58%.

MS m/z (ESI): 375 [M+1]

the fourth step

6-(4-(but-2-ynyl)piperazin-1-yl)-2-(4-phenoxyphenyl)nicotamide

2-(4-Phenoxyphenyl)-6-(piperazin-1-yl)nic acid amide 4c (80 mg, 0.21 mmol) was used as a starting material, which was synthesized by the same procedure as in Example 1 to give the title product 6 -(4-(But-2-ynyl)piperazin-1-yl)-2-(4-phenoxyphenyl)nicotinamide 4 (28 mg, solid), yield: 30%.

MS m/z (ESI): 441 [M+1]

¹ H NMR (400 MHz, CD ₃ OD) δ 7.78 (d, J = 8.7 Hz, 1H), 7.73 - 7.61 (m, 2H), 7.46 - 7.33 (m, 2H), 7.16 (t, J = 7.4 Hz) , 1H), 7.13 - 6.97 (m, 4H), 6.83 (d, J = 8.7 Hz, 1H), 3.92 (dd, J = 6.4, 3.9 Hz, 2H), 3.79 (dd, J = 6.3, 4.0 Hz, 2H), 3.73 (s, 1H), 2.08 (s, 3H).

Example 5

6-(1-(but-2-ynyl)pyrrolidin-3-yl)-2-(4-phenoxyphenyl)nicotinamide

Example 5 was synthesized by following the procedure of Example 4, but in the first step, tert-butyl piperazine-1-carboxylate was replaced with t-butylpyrrolidin-3-ylcarbamate.

MS m/z (ESI): 441 [M+1]

¹ H NMR (400 MHz, CD ₃ OD) δ 7.70 (ddd, J = 11.7, 9.5, 5.7 Hz, 3H), 7.45 - 7.30 (m, 2H), 7.15 (t, J = 7.4 Hz, 1H), 7.03 (ddd, J = 9.5, 6.7, 1.9 Hz, 4H), 6.47 (d, J = 8.7 Hz, 1H), 4.57 - 4.44 (m, 1H), 3.82 (dd, J = 11.1, 6.3 Hz, 1H), 3.65 (ddd, J = 20.2, 12.1, 4.4 Hz, 2H), 3.48 (dd, J = 11.1, 4.4 Hz, 1H), 2.31 (dd, J = 13.4, 6.2 Hz, 1H), 2.13 - 2.01 (m, 1H), 1.97 (s, 3H).

Example 6

6-((1-(but-2-ynyl)pyrrolidin-3-yl)amino)-2-(4-phenoxyphenyl)nicotinamide

Example 6 was synthesized by reference to the procedure of Example 4, except that tert-butyl 3-aminopyrrolidin-1-carboxylate was substituted for tert-butyl piperazine-1-carboxylate in the first step.

MS m/z (ESI): 441 [M+1]

¹ H NMR (400 MHz, CD ₃ OD) δ 7.71 - 7.58 (m, 3H), 7.39 (t, J = 7.7 Hz, 2H), 7.16 (dd, J = 7.4, 6.5 Hz, 1H), 7.10 - 6.96 (m, 4H), 6.52 (dd, J = 8.6, 5.3 Hz, 1H), 4.61 (dd, J = 6.8, 3.3 Hz, 1H), 4.07 (dd, J = 11.3, 5.9 Hz, 1H), 3.93 - 3.72 (m, 2H), 3.70 - 3.37 (m, 2H), 2.39 - 2.24 (m, 1H), 2.11 - 1.94 (m, 4H).

Example 7

1-(1-(but-2-ynyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide

first step

Tert-butyl 3-((methylsulfonyl) oxo)pyrrolidine-1-carboxylate

The compound tert-butyl 3-hydroxypyrrolidine-1-carboxylate 7a (561 mg, 3.0 mmol), methanesulfonyl chloride (410 mg, 3.6 mmol), triethylamine (606 mg, 6.0 mmol) and dichloromethane 50 mL) was mixed and stirred at room temperature for 4 hours. This mixture was diluted with 50 mL of dichloromethane and washed successively with a saturated aqueous solution of ammonium chloride (20 mL) and brine (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, and then filtered, and then evaporated to remove the solvent, and the solvent was evaporated to give the desired product t-butyl 3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate 7b (790 mg). Yield: 99%. The product was used in the next reaction without purification.

MS m/z (ESI): 266 [M+1]

Second step

Ethyl 3-carbonyl-3-(4-phenoxyphenyl)propanoate

The mixture diethyl carbonate (8.8 g, 75 mmol), sodium hydride (60%, 3.0 g, 75 mmol) and toluene (50 mL) were heated to 90 ° C, then the compound 1-(4-phenoxyphenyl)ethanone was added. A solution of 7c (6.36 g, 30 mmol) elute After cooling to room temperature, 50 mL of water was added, the toluene was removed under reduced pressure, and the residue was diluted with water (100 mL) and extracted with dichloromethane (100 mL×3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, evaporated, evaporated, evaporated, evaporated, evaporated Ethyl 3-(4-phenoxyphenyl)propanoate 7d (5.54 g), Yield: 65%.

MS m/z (ESI): 285 [M+1]

third step

Ethyl 3-ethoxy-2-(4-phenoxybenzoyl)acrylate

The compound 3-carbonyl-3-(4-phenoxyphenyl)propanoic acid ethyl ester 7d (5.54g, 19.5mmol), triethyl orthoformate (11mL, 66mmol) and acetic anhydride (28mL) were mixed and heated Stir at 4 ° C for 4 hours. The residue was purified by silica gel column chromatography (ethyl ether / ethyl acetate=50/1) to afford ethyl 3-ethoxy-2-(4-phenoxybenzoyl) 7e (2.6 g), yield: 25%.

MS m/z (ESI): 341 [M+1]

the fourth step

Ethyl 3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylate

The compound 3-ethoxy-2-(4-phenoxybenzoyl) acrylate 7e (2.6 g, 7.6 mmol), hydrazine hydrate (0.39 g, 7.6 mmol) and ethanol (50 mL) were mixed and heated. Stir at 21 ° C for 21 hours. The mixture was cooled to room temperature, and then evaporated to dryness. Ethyl 4-carboxylate 7f (2.0 g), Yield: 85%.

MS m/z (ESI): 309 [M+1]

the fifth step

Ethyl 1-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylate

The compound 3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylic acid ethyl ester 7f (308 mg, 1.0 mmol), 3-((methylsulfonyl)oxy) tert-butyl 3- ((Methanesulfonyl)oxo)pyrrolidine-1-carboxylate 7b (477 mg, 1.8 mmol), cesium carbonate (650 mg, 2.0 mmol) and anhydrous N,N-dimethylformamide (20 mL), Heat to 100 ° C and stir for 16 hours. The mixture was cooled to rt. The organic phase was combined, dried over anhydrous sodium sulfate, filtered, evaporated, evaporated, evaporated, evaporated 1-(tert-Butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylate 7 g (334 mg), Yield: 70% .

MS m/z (ESI): 478 [M+1]

Step 6

1-(1-(tert-Butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylic acid

The mixture ethyl 1-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylate 7g (334mg , 0.7 mmol), an aqueous lithium hydroxide solution (1N, 7 mL, 7.0 mmol) and ethanol (20 mL) were heated to 75 ° C and stirred for 1 hour. After cooling to room temperature, it was taken to dryness under reduced pressure, and the residue was diluted with 5 mL of water and acidified to pH 6 with 2N hydrochloric acid. Filtration, the obtained solid was washed with 15 mL of water and dried in vacuo to give 1-(1-(t-butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H. Pyrazole-4-carboxylic acid 7h (230 mg, solid), yield: 73%.

MS m/z (ESI): 450 [M+1]

Seventh step

Tert-Butyl 3-(4-carbamoyl-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate

Ammonia gas to the compound 1-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxylic acid 7h (230mg , 0.51 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (232 mg, 0.61 mmol) and anhydrous N,N- The mixture of dimethylformamide (10 mL) was bubbled for 30 minutes and the resulting reaction mixture was stirred at room temperature for 30 minutes. It was diluted with 30 mL of ethyl acetate, and washed with water (20 mL × 2) and brine (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, evaporated, evaporated, evaporated, evaporated -carbamoyl-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)pyrrolidine-l-carboxylate 7i (190 mg, oil), yield: 83%.

MS m/z (ESI): 449 [M+1]

Eighth step

3-(4-phenoxyphenyl)-1-(pyrrolidin-3-yl)-1H-pyrazole-4-carboxamide

The compound tert-butyl 3-(4-carbamoyl-3-(4-phenoxyphenyl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate 7i (190 mg, 0.42 mmol The ethanolic solution of hydrochloric acid (4N, 1 mL, 4.0 mmol) and dichloromethane (2 mL) were mixed and stirred at room temperature for 16 hours. Desolvation under reduced pressure gave the title product 3-(4-phenoxyphenyl)-1-(pyrrolidin-3-yl)-1H-pyrazole-4-carboxamide 7j (120 mg, m.

MS m/z (ESI): 349 [M+1]

Step 9

1-(1-(but-2-ynyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide

3-(4-phenoxyphenyl)-1-(pyrrolidin-3-yl)-1H-pyrazole-4-carboxamide 7j (120 mg, 0.34 mmol) was used as a starting material, and reference was made to the compound of Example 1 Synthetic method to give the title product 1-(1-(but-2-ynyl)pyrrolidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide 7 (30 mg, solid), Yield: 21%.

MS m/z (ESI): 415 [M+1]

¹ H NMR (400 MHz, CD ₃ OD) δ 8.18 (d, J = 7.0 Hz, 1H), 7.71 (dd, J = 8.7, 1.9 Hz, 2H), 7.39 (t, J = 7.9 Hz, 2H), 7.15 (t, J = 7.4 Hz, 1H), 7.04 (dd, J = 13.9, 8.3 Hz, 4H), 5.10 (d, J = 2.9 Hz, 1H), 4.22 - 4.12 (m, 1H), 4.06 - 3.86 (m, 2H), 3.83 - 3.62 (m, 1H), 2.60 - 2.44 (m, 2H), 2.05 (d, J = 12.0 Hz, 3H).

Example 8

1-((1-(but-2-ynyl)piperidin-4-yl)methyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide

Example 8 was synthesized by following the procedure of Example 7, but in the first step, tert-butyl 3-hydroxypyrrolidine-1 was replaced with t-butyl 4-(hydroxymethyl)piperidine-1-carboxylate. - a carboxylic acid ester.

MS m/z (ESI): 443 [M+1]

¹ H NMR (400 MHz, CD ₃ OD) δ 8.10 (s, 1H), 7.72 - 7.64 (m, 2H), 7.45 - 7.34 (m, 2H), 7.16 (t, J = 7.4 Hz, 1H), 7.10 – 6.98 (m, 4H), 4.46 (dd, J = 12.7, 8.8 Hz, 2H), 4.12 (d, J = 7.2 Hz, 2H), 3.17 (d, J = 2.5 Hz, 1H), 2.74 (d, J = 2.9 Hz, 1H), 2.35 - 2.19 (m, 1H), 2.04 (s, 3H), 1.71 (d, J = 15.5 Hz, 2H), 1.31 (s, 2H).

Example 9

1-(1-(but-2-ynyl)azetidin-3-yl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide

Example 9 was synthesized by following the procedure of Example 7, but in the first step, tert-butyl 3-hydroxypyrrolidine-1-carboxylate was substituted with tert-butyl 3-hydroxypyrrolidine-1-carboxylic acid. ester.

MS m/z (ESI): 401 [M+1]

^{1 H NMR (400MHz, MeOD)} δ8.21 (s, 1H), 7.75 (d, J = 8.7Hz, 2H), 7.44-7.33 (m, 2H), 7.16 (t, J = 7.4Hz, 1H), 7.11–6.93 (m, 4H), 5.33 (s, 1H), 4.69 (dd, J=20.5, 6.8 Hz, 2H), 4.47 (dd, J=27.8, 6.7 Hz, 2H), 2.05 (s, 3H) .

BTK activity inhibition test

Evaluation of the effects of the compounds of the invention on the activity of Bruton tyrosine kinase (BTK) using an in vitro kinase assay

The experimental methods are summarized as follows:

The in vitro activity of BTK was determined by detecting the level of ADP produced in the kinase reaction using Promega's ADP-Glo Kinase Assay Kit. In a kinase assay, the kinase consumes ATP to phosphorylate the substrate while producing ADP. The ADP-Glo reagent will stop the kinase reaction and completely consume the remaining ATP, and finally add the kinase detection reagent to convert the ADP to a new ATP. The luciferase in the detection reagent can catalyze fluorescein with the participation of ATP and O ₂ to produce oxidized fluorescein, AMP and generate photons, thereby converting the chemical signal into an optical signal (Luminecence). The intensity of the light signal is positively correlated with the amount of ADP produced in the kinase reaction, thereby enabling quantitative detection of the activity of the kinase BTK.

All assays were performed at 23 ° C with constant temperature, using Corning 3674 white 384-well assay plate, kinase BTK (full length His-Tag) purified by in-house expression, and the kinase substrate was peptide (4:1 Glu, Tyr) (purchased from SignalChem) And ATP (Sigma), the optical signal was read using a microplate reader EnVision (Perkin Elmer). The assay buffer included 40 mM Tris-HC (pH 7.5), 10 mM MgCl ₂ (Sigma), 2 mM MnCl ₂ (Sigma), 0.05 mM DTT (Sigma), and 0.01% BSA (Sigma); the kinase BTK was prepared using assay buffer as A kinase reaction solution having a concentration of 1.6 ng/uL; the substrate reaction solution includes 0.2 mg/mL of polypeptide substrate and 50 μm of ATP.

The compound IC ₅₀ was calculated from the 10 concentration points by the following formula. Compounds were first diluted in 100% DMSO to 1 mM, then serially diluted 3 fold with DMSO to a minimum concentration of 0.05 μm, and each concentration was diluted 40-fold with assay buffer. Add 1 uL series of compound solution and 2 uL of kinase reaction solution to the 384-well assay plate, mix well and incubate at room temperature for 30 minutes in the dark; then add 2 uL of substrate reaction solution, the total volume of the reaction is 5 uL, and the reaction mixture is avoided at room temperature. light for 60 minutes; followed by addition of the reaction volume and the like 5uL ADP-Glo ^TM reagent termination of the reaction, the mixed stand at room temperature 40 minutes; finally adding 10uL kinase Assay dark at room temperature for 30 minutes and then read on an Envision value .

The percent inhibition is calculated based on the following formula:

Inhibition %=[1-(RLU _compound -RLU _min )/(RLU _max -RLU _min )]X100

Where RLU _compounds are cold light readings at a given compound concentration, RLU _min is a cold light reading without the addition of a kinase, and RLU _max is a cold light reading without the addition of a compound. By using Excel XLfit program to calculate the IC ₅₀ of the compound.

Compound number	IC 50 (nM)	Compound number	IC 50 (nM)
1.	A	2.	A
3.	C	4.	A
5.	A	6.	A
7.	A	8.	C
9.	A

A<100nM; B=100 to 500nM; C>500nM

Conclusion: The compounds of the present invention have a significant inhibitory effect on the activity of Bruton tyrosine kinase (BTK).

Claims (9)

1. a compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable compound thereof Salt used:

   

   among them:

   Rings A and B ¹ are each independently selected from aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted by one or more G ¹ ;

   B ^{2 is} independently selected from H, C _3-8 cyclo, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein the cyclo, heterocyclyl, aryl or heteroaryl is optionally Or replaced by multiple G ² ;

   L ^{1 is} independently selected from -C _0-2 alkyl-, -CR ¹ R ² -, -C _1-2 alkyl(R ¹ )(OH)-, -C(O)-, -CR ¹ R ² O-, -OCR ¹ R ² -, -SCR ¹ R ² -, -CR ¹ R ² S-, -NR ¹ -, -NR ¹ C(O)-, -C(O)NR ¹ -, -NR ¹ CONR ² -, -CF ₂ -, -O-, -S-, -S(O) _m -, -NR ¹ S(O) _m - or -S(O) _m NR ¹ -;

   L ^{2 is} independently selected from -C _0-4 alkyl-, -C(O)-, -O-, -NR ³ -, -NR ³ C(O)- or -NR ³ S(O) _m -;

   X is independently selected from C _0-4 alkyl, C _3-8 cyclo, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein the alkyl, cyclo, heterocyclyl, aryl or The heteroaryl group is optionally substituted with one or more G ³ ;

   Y is independently selected from -C(O)-, -NR ⁴ C(O)-, -S(O) _m - or -NR ⁴ S(O) _m -;

   R is independently selected from H, D, alkyl, cyclo, heterocyclyl, aryl or heteroaryl, wherein the alkyl, cyclo, heterocyclyl, aryl or heteroaryl is optionally taken by one or Replaced by multiple G ⁴ ;

   R ¹ , R ² , R ³ and R ⁴ are each independently selected from H, D, C _1-8 alkyl, C _3-8 cyclic, 3-8 membered heterocyclyl, aryl or heteroaryl, wherein The alkyl, cyclo, heterocyclyl, aryl or heteroaryl group is optionally substituted by one or more G ⁵ ;

   G ¹ , G ² , G ³ , G ⁴ and G ⁵ are each independently selected from H, D, halogen, cyano, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl , -OR ⁵ , -OC(O)NR ⁵ R ⁶ , -C(O)OR ⁵ , -C(O)NR ⁵ R ⁶ , -C(O)R ⁵ , -NR ⁵ R ⁶ , -NR ⁵ C(O)R ⁶ , -NR ⁵ C(O)NR ⁶ R ⁷ , -S(O) _m R ⁵ or -NR ⁵ S(O) _m R ⁶ , wherein the alkyl, alkenyl, alkynyl group Or a cycloalkyl, heterocyclyl, aryl or heteroaryl group optionally selected from one or more selected from the group consisting of D, halogen, cyano, C _1-8 alkyl, C _3-8 ring, 3-8 membered heterocyclic ring Base, -OR ⁷ , -OC(O)NR ⁷ R ⁸ , -C(O)OR ⁷ , -C(O)NR ⁷ R ⁸ , -C(O)R ⁷ , -NR ⁷ R ⁸ , -NR Substituting a substituent of ⁷ C(O)R ⁸ , —NR ⁷ C(O)NR ⁸ R ⁹ , —S(O) _m R ⁷ , —NR ⁷ S(O) _m R ⁸ ;

   R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ are each independently selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-8 cycloalkyl, 3-8 membered monocyclic hetero a cyclic group, a monocyclic heteroaryl group or a monocyclic aryl group;

   m is 1 or 2.
2. A compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof according to claim 1 a mixture form, and a pharmaceutically acceptable salt thereof, which is a compound of the formula (II) or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer Constructs, mixtures thereof, and pharmaceutically acceptable salts thereof:

   

   among them:

   Ring B is

   

   Z ¹ and Z ² are each independently selected from CH or N;

   Z ³ is O or S;

   The definitions of B ¹ , B ² , L ¹ , L ² , X, Y and R are as set forth in claim 1.
3. A compound of the formula (I) according to any one of claims 1 to 2, or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof And a mixture thereof, and a pharmaceutically acceptable salt thereof, which is a compound of the formula (III) or a tautomer thereof, a mesogen, a racemate, an enantiomer, a non- Enantiomers, mixtures thereof, and pharmaceutically acceptable salts thereof:

   

   among them:

   B ¹ and B ² are a benzene ring or a six-membered heteroaryl ring;

   Rings B, L ² , X, Y, R, G ¹ and G ² are as defined in claims 1-2.
4. The compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof according to any one of claims 1 to 3 And mixtures thereof, and pharmaceutically acceptable salts thereof, which are compounds of the formula (IV) or tautomers, meso, racemates, enantiomers, non- Enantiomers, mixtures thereof, and pharmaceutically acceptable salts thereof:

   

   among them:

   P ¹ and P ² are each independently selected from C(R ^a ) or N;

   R ^a is H or an alkyl group;

   n and o are each independently selected from 0, 1 or 2;

   Rings B, B ¹ , B ² , L ² , Y and R are as defined in claims 1-3.
5. A compound of the formula (I) or a tautomer, a mesogen, a racemate, an enantiomer or a diastereomer thereof according to any one of claims 1 to 4 And mixtures thereof, and pharmaceutically acceptable salts thereof, wherein R is independently selected from H, D, alkyl or heteroalkyl.
6. The compound of the formula (I) according to claim 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer thereof, or a mixture thereof And a pharmaceutically acceptable salt thereof, wherein the compound is:

   
7. A pharmaceutical composition comprising a therapeutically effective amount of a compound of the formula (I) according to any one of claims 1 or a tautomer, a mesogen thereof, a foreign body The rotators, enantiomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers, diluents and excipients.
8. The compound of the formula (I) or a tautomer, a mesomer, a racemate, an enantiomer or a diastereomer thereof, according to any one of claims 1 A mixture thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7 for use in the preparation of a Bruton tyrosine kinase (BTK) inhibitor.
9. The compound of the formula (I) or a tautomer, a mesomer, a racemate, an enantiomer or a diastereomer thereof, according to any one of claims 1 A mixture thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8 for use in the preparation of a medicament for treating and/or preventing a disease such as a tumor and inflammation.