CN111961035A - A class of compounds containing hydroxyisoquinoline structures, pharmaceutical compositions and applications thereof - Google Patents

Abstract

The invention relates to a compound containing a hydroxyisoquinoline structure, a pharmaceutical composition and application thereof, wherein the compound has a structure shown in a formula (I). The compound with the structure shown as the formula (I) has good inhibition effect on the activity of Bruton kinase, and the half inhibition concentration of the compound is generally 10^‑7mol.L^‑1The following. Meanwhile, the compound with the structure of the formula (I) prepared in the embodiment of the invention shows specific anti-inflammatory activity on different animal models.

Description

A kind of compound containing hydroxyisoquinoline structure, pharmaceutical composition and its application

technical field

The invention relates to the field of medicinal chemistry, in particular to a class of compounds containing hydroxyisoquinoline structures, pharmaceutical compositions and applications thereof.

Background technique

Protein kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by adding phosphate groups to proteins (T. Hunter, Cel 1987 50:823-829). In particular, tyrosine kinases phosphorylate proteins on the phenolic moieties of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration and differentiation. Aberrant kinase activity has been implicated in many human diseases, including cancer, autoimmune and inflammatory diseases. Because protein kinases are among the key regulators of cell signaling, they provide targets for modulating cellular function with small molecule kinase inhibitors, and are therefore good targets for drug design. In addition to the treatment of kinase-mediated disease processes, selective and potent inhibitors of kinase activity can be used to study cellular signaling processes and to identify other therapeutically meaningful cellular targets.

There is good evidence for the critical role of B cells in the pathogenesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics that deplete B cells, such as Rituxan, are effective against autoantibody-induced inflammatory diseases such as rheumatoid arthritis (Rastetter et al., Annu Rev Med 2004 55:477). Therefore, inhibitors of protein kinases that play a role in B cell activation should be useful therapeutics for B cell mediated disease pathologies such as autoantibody production.

Signaling through the B-cell receptor (BCR) controls a range of B-cell responses, including proliferation and differentiation into mature antibody-producing cells. The BCR is a key regulator of B cell activity and aberrant signaling can lead to dysregulated B cell proliferation and the formation of pathogenic autoantibodies, which contribute to a variety of autoimmune and/or inflammatory diseases. Bruton's tyrosine protein kinase (Btk) is a non-BCR-associated kinase that is membrane-proximal and immediately downstream of the BCR. Deficiency of Btk has been shown to block BCR signaling, and thus inhibition of Btk may be an effective therapeutic approach to block B cell-mediated disease processes.

Btk is a member of the Tec family of tyrosine kinases and has been shown to be a key regulator of early B cell formation as well as activation and survival of mature B cells (Khan et al. Immunity 1995 3:283; Ellmeier et al. J.Exp.Med.2000192: 1611). Btk mutations in humans cause the disorder X-linked agammaglobulinemia (XLA) (reviewed in Rosen et al. New Eng. J. Med. 1995 333:431 and Lindvall et al. Immunol. Rev. 2005 203:200). These patients were immunocompromised and displayed impaired B cell maturation, reduced immunoglobulin and topical B cell levels, reduced T cell-independent immune responses, and attenuated calcium mobilization following BCR stimulation.

Evidence for the role of Btk in autoimmune and inflammatory diseases has been provided by Btk-deficient mouse models. In a preclinical murine model of systemic lupus erythematosus (SLE), Btk-deficient mice showed marked improvement in disease progression. Furthermore, Btk-deficient mice are resistant to collagen-induced arthritis (Jansson and Holmdahl Clin. Exp. Immunol. 1993 94:459). Dose-dependent efficacy of selective Btk inhibitors in a mouse arthritis model has been demonstrated (z. Pan et al. Chem. Med Chem. 2007 2:58-61).

Btk is also expressed by cells other than B cells that may be involved in disease processes. For example, Btk is expressed by mast cells and Btk-deficient bone marrow-derived mast cells show impaired antigen-induced degranulation (Iwaki et al. J. Biol. Chem. 2005 280:40261). This shows that Btk can be used to treat pathological mast cell responses such as allergy and asthma. Furthermore, monocytes from XLA patients in which Btk activity is lacking show reduced TNFα production following stimulation (Horwood et al. J Exp Med 197:1603, 2003). Therefore, TNFα-mediated inflammation can be modulated by small molecule Btk inhibitors. In addition, Btk has been reported to play a role in apoptosis (Oslam and Smith Immunol. Rev. 2000 178:49), and thus Btk inhibitors would be effective for the treatment of certain B-cell lymphomas and leukemias (Feldhahn et al. JExp. Med. 2005 201:183 7).

Although a series of Bruton's kinase inhibitors have been disclosed, there is still a need to develop new compounds with richer structural types, which may have higher activity and better druggable properties.

SUMMARY OF THE INVENTION

Through continuous efforts, the present invention devised a compound having the structure represented by the general formula (I), and found that the compound having such a structure exhibited excellent effects and functions.

The application provides a Btk inhibitor compound whose structure is shown in formula (I), and its method of use, as described herein below:

The compound whose structure is shown in formula (I) or a pharmaceutically acceptable salt thereof:

in,

X, Y, Z, W are independently selected from CH or N;

Ring A is C ₆ -C ₁₀ aryl, C ₆ -C ₁₀ heteroaryl;

Ring B is C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl;

R ₁ , R ₂ are selected from H, halogen, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl, Said alkyl, alkoxy, cycloalkyl, heterocycloalkyl are optionally substituted by one or more halogens, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl substitution;

R ₃ is selected from

Further, Z is selected from CH, and W is selected from N or CH; or Z is selected from N, and W is selected from CH.

Further, ring A is a benzene ring or a pyridine ring.

Further, ring B is cyclohexane, cyclopentane, piperidine, morpholine, and pyrrolidine.

Further, the compound shown in formula (I) or a pharmaceutically acceptable salt thereof, at least one such as formula (II),

Z is selected from CH, W is selected from N or CH; or Z is selected from N, W is selected from CH;

Ring A is a benzene ring, a pyridine ring;

Ring B is cyclohexane, cyclopentane, piperidine, morpholine, pyrrolidine;

R ₁ , R ₂ are selected from H, halogen, cyano, C ₁ -C ₄ alkyl, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl, Said alkyl, alkoxy, cycloalkyl, heterocycloalkyl are optionally substituted by one or more halogens, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₆ heterocycloalkyl substitution;

R ₃ is selected from

Further, the compound is selected from:

Use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a BTK inhibitor.

Such BTK inhibitors can be used for use in medicines for the treatment of inflammatory and/or autoimmune disorders, particularly rheumatoid arthritis.

To test the level of action of the compounds provided by the present invention on protein kinases, biochemical level enzyme activity assays and cellular level enzyme activity assays are used to determine the activity and level of action of various compounds of the present invention on one or more PKs. Similar experiments can be designed in the same manner for any kinase using methods well known in the art.

In the enzymatic activity test at the biochemical level, the activity of tyrosine kinases is detected by HTRF technology, a time-resolved fluorescence resonance energy transfer technology, which can be performed according to known instructions or literature methods, see Kolb et al., "Tyrosine kinase assays adapted to homogenous time-resolved fluorescence”. Drug Discovery Today. Vol 3: pp 333-342. HTRF (Homogeneous Time-Resolved Fluorescence) is one of the most commonly used methods to detect analytes in homogeneous systems. This technique combines fluorescence resonance energy transfer (FRET) and time-resolved technology (TR), and has been widely used. Applied to different stages of drug development based on cell experiments and biochemical experiments. According to the determination principle of HTRF method, after incubating the pure enzyme with biotinylated substrate and ATP, avidin-labeled XL-665 and Eu-labeled antibody recognizing the phosphorylation of the substrate were added. After phosphorylation, the Eu-labeled antibody can recognize the phosphorylated product and form time-resolved fluorescence resonance energy transfer (FRET) with avidin-labeled XL665, while the unphosphorylated substrate cannot be recognized by the antibody. The FRET signal was formed, and the inhibitory activity of the analyte on Btk tyrosine kinase at different concentrations was determined by measuring the difference between the fluorescence signals at 665 nm and 620 nm. Thus, the activity of the compounds of the invention on biochemical levels of Btk tyrosine kinase can be determined using this method, while similar assays can be used for other protein kinases using methods well known in the art.

The determination of enzymatic activity at the cellular level is achieved by measuring calcium flux. Fluo-4Direct ^™ Calcium Assay Kits were used for this experiment. The main dye used in its kit is Fluo 4-AM. Fluo 4-AM is an acetyl methyl ester derivative of Fluo4 that can easily enter cells through culture. After AM enters the cell, it will be hydrolyzed by intracellular esterase, and the resulting Fluo 4 will then combine with calcium ions and emit fluorescence. The changes in intracellular calcium ion concentration can be detected by instruments such as laser confocal microscopy or flow cytometry.

The pharmacokinetic properties of the compounds in rats can also be investigated by adopting the general in vivo pharmacokinetic test method in rats.

The general mouse Arthus Reaction model or the rat collagen-induced arthritis (rCIA) model can be used to examine the in vivo efficacy of the compounds in mice or rats.

The compound prepared by the invention with the structure shown in formula (I) has a good inhibitory effect on Bruton kinase activity, and its half inhibitory concentration (IC ₅₀ ) is generally below 10 ^-7 mol/L. Meanwhile, the compounds with the structure of formula I prepared in the examples of the present invention have good oral pharmacokinetic properties, and show clear in vivo pharmacokinetics in Arthus Reaction model or rat collagen-induced arthritis (rCIA) model. It is inferred that the compound of the present invention having the structure of formula (I) can be applied to prepare a medicament for treating Bruton kinase-related inflammatory and/or autoimmune disorders in organisms.

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

"Alkyl" refers to a saturated aliphatic hydrocarbon group. Straight or branched chain groups comprising 1 to 20 carbon atoms. Preferred are medium sized alkyl groups containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, and the like. More preferred are lower alkyl groups containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or tert-butyl, and the like. Alkyl may be substituted or unsubstituted, and when substituted, preferred groups are: halogen, C2 _{- C6} alkenyl, _C6 - _C10 aryl, _C5 - _C10 heteroaryl, halogen Substituted C ₁ -C ₆ alkyl, 4- to 8-membered heteroalicyclic, hydroxyl, C ₁ -C ₆ alkoxy, C ₆ -C ₁₀ aryloxy.

"Alkylene" means a divalent saturated straight hydrocarbon group of 1 to 10 carbon atoms (eg (CH ₂ ) _n ) or a branched saturated divalent hydrocarbon group of 2 to 10 carbon atoms (eg -CHMe), unless otherwise stated . Except in the case of methylene, the open state of an alkylene group is not attached to the same atom. Examples of alkylene groups include, but are not limited to: methylene, ethylene, propylene, 2-methyl-propylene, 1,1-dimethyl-ethylene, butylene, 2-ethyl butylene.

"Cycloalkyl" refers to a 3- to 8-membered all-carbon monocyclic, all-carbon 5-/6- or 6-/6-membered fused ring or polycyclic fused ring ("fused" ring means that each of the (rings share an adjacent pair of carbon atoms with other rings in the system) groups in which one or more of the rings have a fully attached pi electron system, examples (without limitation) of cycloalkyl groups are cyclopropane, cyclobutane, Cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane and cycloheptatriene. Cycloalkyl is substitutable and substituted. When substituted, the substituents are preferably one or more groups each selected from the group consisting of: hydrogen, hydroxy, mercapto, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkane group, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylene, aryl, heteroaryl, alkoxycarbonyl, Amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, Alkylcarbonylamino, arylcarbonylamino.

"Aryl" means an all-carbon monocyclic or fused polycyclic group of 6 to 14 carbon atoms with a fully conjugated pi electron system. "Aryl" includes:

Six-membered carbon aromatic rings, such as benzene;

Bicyclic rings, at least one of which is a carboaromatic ring, such as naphthalene, indene and 1,2,3,4-tetrahydroquinoline; and

Tricyclic rings, at least one of which is a carboaromatic ring, eg, fluorene.

For example, aryl includes a six-membered carboaromatic ring and a six-membered heterocycle containing one or more heteroatoms selected from nitrogen, oxygen, and sulfur, provided that the point of attachment is on the carboaromatic ring. Aryl, however, does not include nor overlap in any way with heterocyclic aryl groups as separately defined below. Thus, as defined herein, if one or more carboaromatic rings are concatenated with a heteroaromatic ring, the resulting ring system is a heteroaryl group, not an aryl group. Non-limiting examples of aryl groups are phenyl, naphthyl. Aryl groups can be substituted or unsubstituted. When substituted, the preferred groups are: hydrogen, hydroxy, nitro, cyano, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkane Sulfanyl, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkyl Sulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonyl amino.

"Heteroaryl" means a monocyclic or fused ring group of 5 to 14 ring atoms, containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C , additionally has a fully conjugated π-electron system. Heteroaryl refers to:

5-8 membered monocyclic aromatic hydrocarbon containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, other atoms in the ring is a carbon atom;

8-12 membered bicyclic aromatic hydrocarbons containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, the other atoms in the ring are carbon atoms; at least one of the rings is aromatic; and

11-14 membered tricyclic aromatic hydrocarbons containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, other atoms in the ring are carbon atoms; at least one of the rings is aromatic.

For example, a heteroaryl group includes a 5-6 membered heteroaromatic ring and a 5-6 membered cycloalkyl group. For such bicyclic juxtaposed heteroaryl groups in which only one ring contains one or more heteroatoms, the point of attachment is on the heteroaromatic ring.

When the total number of sulfur atoms and oxygen atoms on the heteroaryl group exceeds 1, these heteroatoms will not be adjacent one by one. In some embodiments, the total number of sulfur atoms and oxygen atoms in a heteroaryl group does not exceed 2. In some embodiments, the total number of sulfur atoms and oxygen atoms in a heteroaryl group does not exceed one.

W选自O、S或NR¹²，各基团如前所述，实例还可以是双环的，诸如，例如，3，8-二氮杂-双环[3.2.1] 辛烷、2，5-二氮杂双环[2.2.2]辛烷或八氢-吡嗪并[2，1-c][1,4]噁嗪。其杂环烷基 (和衍生物)包括其离子形式。Examples of heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, triazole, pyrimidine, pyridine, pyridone, imididine, pyrazine, pyridazine, indole, Azaindole, benzimidazole, benzotriazole, indoline, indolinone, quinoline, isoquinoline, quinazoline, thienopyridine, thienopyrimidine, etc. Preferred examples of such groups are pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furyl, oxazolyl, thienyl, thiazolyl, benzimidazolyl, benzotriazole. One or all of the hydrogen atoms in the heteroaryl group may be replaced by the following groups: hydrogen, hydroxy, nitro, cyano, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, Lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, Alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkyl Carbonylamino, arylcarbonylamino. "Heterocycloalkyl" means a monovalent saturated cyclic group consisting of one or more rings, preferably 1 to 2 rings (including spiro ring systems), of 3 to 8 atoms per ring, combined with one or more Multiple ring heteroatoms (selected from N, O or S(O) _0-2 ), and which may be optionally substituted independently by one or more, preferably 1 or 2 substituents, the substituents are selected From: hydrogen, hydroxyl, mercapto, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl , lower cycloalkylalkylene, lower heterocycloalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl , arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino. unless otherwise indicated. Examples of heterocycloalkyl include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepinyl, oxetanyl, tetrahydrofuranyl , tetrahydrothienyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydropyranyl, thiomorpholinyl, quinuclidinyl and imidazolinyl, preferably

W is selected from O, S or ^NR12 , each group is as previously described, examples may also be bicyclic, such as, for example, 3,8-diaza-bicyclo[3.2.1]octane, 2,5- Diazabicyclo[2.2.2]octane or octahydro-pyrazino[2,1-c][1,4]oxazine. Its heterocycloalkyl (and derivatives) include its ionic form.

"Alkoxy" means -O-(unsubstituted alkyl) and -O(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

"Aryloxy" means -O-aryl and -O-heteroaryl. Representative examples include, but are not limited to, phenoxy, pyridyloxy, furanoxy, thienyloxy, pyrimidinyloxy, pyrazinoxy, and the like, and derivatives thereof.

"Arylalkylene" means an alkyl group, preferably a lower alkyl group as defined above, substituted with an aryl group as defined above, eg -CH2phenyl, -( _{CH2 ) 2phenyl} , -( _CH2phenyl ) ₃ phenyl, CH ₃ CH(CH ₃ )CH ₂ phenyl and derivatives thereof.

"Heteroarylalkylene" means an alkyl group, preferably a lower alkyl group as defined above, substituted with a heteroaryl group as defined above, eg -CH2pyridyl, -( _{CH2 ) 2pyrimidinyl} , -( CH ₂ ) ₃ imidazolyl and its derivatives.

"Hydroxy" means the -OH group.

"Sulfhydryl" means the -SH group.

"Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

"Haloalkyl" means an alkyl group, preferably a lower alkyl group as defined above, which is substituted with one or more _of the same or different halogen atoms, eg _-CH2Cl , _-CF3 , _-CCl3 , _-CH2CF3 , -CH ₂ CCl ₃ , etc.

"Cyano" means a -CN group.

"Amino" means the _-NH2 group.

"Nitro" means the -NO ₂ group. "Optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes that the thing or circumstance may or may not occur, and that the event or circumstance occurs and does not occur two cases.

In some embodiments, "substituted with one or more groups" means that one, two, three, or four hydrogen atoms in the specified atom or group, respectively, are selected from the specified range of groups Same or different group substitutions.

Wavy lines indicate attachment sites;

"Pharmaceutically acceptable salts" refer to those salts that retain the biological effectiveness and properties of the parent compound. Such salts include:

(1) Forming a salt with an acid, obtained by the reaction of the free base of the parent compound with an inorganic acid or an organic acid. The inorganic acid includes hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid, etc. , organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid Formic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid, etc. .

(2) The acid proton present in the parent compound is replaced by a metal ion or a salt formed by complexation with an organic base, such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion, an organic base such as ethanolamine, diethanolamine, triethanolamine Ethanolamine, tromethamine, N-methylglucamine, etc.

"Pharmaceutical composition" refers to mixing one or more of the compounds of the present invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, with another chemical ingredient, such as a pharmaceutically acceptable carrier, . The purpose of a pharmaceutical composition is to facilitate the process of administration to an animal.

"Pharmaceutically acceptable carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound, such as, but not limited to: calcium carbonate, calcium phosphate, various Sugar (eg lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic or methacrylic acid polymers, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol Glycol, castor oil or hydrogenated castor oil or polyethoxy hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

In the aforementioned pharmaceutical composition, in addition to a pharmaceutically acceptable carrier, it may also include adjuvants commonly used in pharmacy (agents), such as: antibacterial agents, antifungal agents, antimicrobial agents, preservatives, conditioning agents. Colorants, solubilizers, thickeners, surfactants, complexing agents, proteins, amino acids, fats, carbohydrates, vitamins, minerals, trace elements, sweeteners, pigments, flavors or their combinations, etc.

Detailed ways

The following examples are used to further describe the present invention, but these examples do not limit the scope of the present invention.

Example 1: N-(1-(8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquine Preparation of olin-6-yl)piperidin-3-yl)acrylamide (EXP1)

1) Preparation of 6-chloro-8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)isoquinolin-1(2H)-one (compound C)

8-Bromo-6-chloroisoquinolin-1(2H)-one (2590 mg, 10 mmol), 5-(4-methylpiperazin-1-yl)pyridin-2-amine (1923 mg, 10 mmol), Pd (dppf) ₂ Cl ₂ ·CH ₂ Cl ₂ (2340 mg, 2.3 mmol) was placed in a 100 mL three-necked flask, replaced with argon three times, added 1,4-dioxane (50 mL), stirred for 5 minutes, and added 2M Na ₂ CO ₃ (2700 mg, 25.5 mmol) was replaced with argon for three times, then heated to 100 ° C for 2 hours, the reaction was stopped and cooled to room temperature, and the filtrate was added with water, ethyl acetate (100 mL* 3) Extraction, washed once with saturated NaCl (25 mL), and finally the ester layer was dried with anhydrous Na ₂ SO ₄ and separated by column chromatography to obtain compound C (925 mg, 2.5 mmol) with a yield of 25%. MS(ESI) m/z: [M+H] ⁺ =370.5. ¹ H-NMR (DMSO-d6, 400MHz): δ11.84(s, 1H), 10.05(s, 1H), 7.71(m, 1H), 7.46(s, 1H), 7.13-7.18(m, 2H) , 6.79(d, 1H), 6.61(d, 1H), 6.21(d, 1H), 3.10-3.17(m, 4H), 2.30-2.37(m, 4H), 2.21(s, 3H) ppm.

2) 1-(8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl ) preparation of piperidin-3-yl tert-butyl) carbamate (compound D)

6-Chloro-8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)isoquinolin-1(2H)-one (738 mg, 2 mmol), piperidine- tert-Butyl 3-ylcarbamate (1001 mg, 5 mmol), Pd(OAc) ₂ (100 mg, 0.44 mmol), potassium acetate (980 mg, 100 mmol) were dissolved in 100 mL of anhydrous 1,4-dioxane, under nitrogen protection , heated to reflux, and reacted for 12 hours. The reaction was stopped and cooled to room temperature, filtered through celite, and the filtrate was extracted with water and ethyl acetate (150 mL*3), washed once with saturated NaCl (25 mL), and finally the ester layer was dried with anhydrous Na ₂ SO ₄ Compound D (960 mg, 1.8 mmol) was obtained after chromatographic separation.

3) N-(1-(8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquinoline- Preparation of 6-yl)piperidin-3-yl)acrylamide (EXP1)

1-(8-((5-(4-Methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl) Piperidin-3-yl tert-butyl)carbamate (Compound D) (534 mg, 1 mmol), 0.1 mL of hydrochloric acid, and 50 mL of dichloromethane were added to a 100 mL eggplant flask, and the reaction was carried out at room temperature for 12 hours under nitrogen protection. After the reaction, the aqueous phase was extracted with dichloromethane (50 mL*3). The organic phases were combined and washed three times with pure water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and used directly for the next reaction.

To the above concentrated solution was added 50 mL of anhydrous dichloromethane, triethylamine (303 mg, 3 mmol), acryloyl chloride (180 mg, 2 mmol), under nitrogen protection, and reacted at 45° C. for 4 hours. After the reaction, the reaction solution was concentrated to dryness, and after column chromatography, N-(1-(8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1- Oxo-1,2-dihydroisoquinolin-6-yl)piperidin-3-yl)acrylamide (146 mg, 0.3 mmol), yield: 30%.

MS(ESI) m/z: [M+H] ⁺ =488.4. ¹ H-NMR (DMSO-d6, 400MHz): δ12.01(s, 1H), 10.04(s, 1H), 8.45(m, 1H), 7.69-7.72(m, 1H), 7.13-7.18(m, 1H), 6.65-6.79(m, 2H), 6.42-6.51(m, 2H), 6.04-6.17(m, 2H), 5.74(m, 1H), 3.54(m, 1H), 3.42(m, 1H) , 3.08-3.1 9(m, 3H), 2.32-2.37(m, 4H), 2.18(s, 3H), 1.42-1.57(m, 4H), ppm.

Example 2: N-(1-(8-oxo-1-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-7,8-dihydro-2, Preparation of 7-Naphthyridin-3-yl)pyrrolidin-3-yl)acrylamide (EXP2)

Referring to the method of Example 1, N-(1-(8-oxo-1-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-7,8-dihydro -2,7-Naphthyridin-3-yl)pyrrolidin-3-yl)acrylamide (EXP2) 35 mg, yield: 25%. MS(ESI) m/z: [M+H] ⁺ =473.50. ¹ H-NMR (DMSO-d6, 400MHz): δ 11.78(s, 1H), 10.52(s, 1H), 8.34(s, 1H ), 7.82(d, 1H), 7.45(d, 2H), 7.05(d, 2H), 6.54(t, 1H), 6.2 8(s, 1H), 6.12(m, 1H), 5.85(m, 1H ), 5.72(m, 1H), 3.65(m, 1H), 3.13-3.52(m, 4H), 2.65-2.69(m, 2H), 2.45-2.51(m, 4H), 1.65-1.94(m, 4H) )ppm.

Example 3: N-(1-(5-oxo-4-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-5,6-dihydro-1, Preparation of 6-naphthalen-2-yl)pyrrolidin-3-yl)acrylamide (EXP3)

Referring to the method of Example 1, N-(1-(5-oxo-4-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-5,6- Dihydro-1,6-naphthalen-2-yl)pyrrolidin-3-yl)acrylamide (EXP3) 65 mg, yield: 35%. MS(ESI) m/z: [M+H] ⁺ =473.2. ¹ H-NMR (DMSO-d6, 400MHz): δ 11.82(s, 1H), 11.55(s, 1H), 8.35(s, 1H ), 7.84(d, 1H), 7.47(d, 2H), 7.08(d, 2H), 6.47(m, 1H), 6.08-6.15(s, 2H), 5.76(m, 1H), 3.67( m, 1H), 3.08-3.45 (m, 4H), 2.65 (m, 2H), 2.52-2.55 (m, 6H), 1.67-1.69 (m, 6H) ppm.

Example 4: N-(1-(5-oxo-4-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-5,6-dihydropyrido[ Preparation of 4,3-d]pyrimidin-2-yl)pyrrolidin-3-yl)acrylamide (EXP4)

Referring to the method of Example 1, N-(1-(5-oxo-4-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-5,6- Dihydropyrido[4,3-d]pyrimidin-2-yl)pyrrolidin-3-yl)acrylamide (EXP4) 28 mg, yield: 29%. MS (ESI) m/z: [M+H] ⁺ =474.3. ¹ H-NMR (DMSO-d6, 400 MHz): δ 11.78 (s, 1H), 8.83 (s, 1H), 8.34 (s, 1H) ), 7.54(d, 1H), 7.38(d, 2H), 7.05(d, 2H), 6.45(m, 1H), 6.0 8(m, 1H), 5.93(d, 1H), 5.75(m, 1H) ),3.69(m,1H),3.45(m,1H),3.14-3.19(m,2H),3.05(m,1H),2.68(m,2H),2.53(m,4H),1.64-1.69( m,6H)ppm.

Example 5: N-(1-(8-((4-(2-methoxyethoxy)phenyl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl ) Preparation of pyrrolidin-3-yl)acrylamide (EXP5)

Referring to the method of Example 1, N-(1-(8-((4-(2-methoxyethoxy)phenyl)amino)-1-oxo-1,2-dihydroisoquine was prepared Lin-6-yl)pyrrolidin-3-yl)acrylamide (EXP5) 65 mg, yield: 12%. MS (ESI) m/z: [M+H] ⁺ =448.6. ¹ H-NMR (DMSO-d6, 400MHz): δ 12.36(s, 1H), 11.85(s, 1H), 8.33(s, 1H ), 7.68(d, 1H), 7.42(d, 2H), 6.95(d, 2H), 6.43-6.45(m, 3H), 6.08-6.18(m, 2H), 5.75(d, 1H), 4.32( m, 2H), 3.78 (m, 2H), 3.40-3.44 (m, 4H), 3.05-3.19 (m, 3H), 1.64-1.69 (m, 2H) ppm.

Example 6: N-(1-(8-((5-(4-Methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquinoline Preparation of olin-6-yl)pyrrolidin-3-yl)acrylamide (EXP6)

Referring to the method of Example 1, N-(1-(8-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)-1-oxo-1,2 was prepared -Dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP6) 74 mg, yield: 12%. MS(ESI) m/z: [M+H] ⁺ =474.3. ¹ H-NMR (DMSO-d6, 400MHz): δ 12.03(s, 1H), 10.07(s, 1H), 8.37(m, 1H), 7.74(m, 1H), 7.19(m, 1H), 6.79 (m,1H),6.67(m,1H),6.45(m,3H),6.07-6.16(m,2H),5.78(m,1H),3.68(m,1H),3.05-3.44(m,4H) ), 2.38(m, 4H), 2.18(m, 3H), 1.67-1.92(m, 2H) ppm.

Example 7: N-(1-(8-((5-((S)-3-methyl-4-(oxetan-3-yl)piperazin-1-yl)pyridine-2- yl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl)pyrrolidin-3-yl)but-2-ynamide (EXP7) preparation

Referring to the method of Example 1, N-(1-(8-((5-((S)-3-methyl-4-(oxetan-3-yl)piperazin-1-yl) was prepared )pyridin-2-yl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl)pyrrolidin-3-yl)but-2-ynamide (EXP7) 82mg, yield: 35%. MS(ESI) m/z: [M+H] ⁺ =542.5. ¹ H-NMR (DMSO-d6, 400MHz): δ11.91(s,1H), 10.01(s,1H), 8.09(s,1H), 7.68(d,1H), 7.18(s,1H), 6.79 (d,1H),6.68(d,1H),6.44(m,2H),6.18(m,1H),4.68-4.82(m,4H),3.67-3.85(m,2H),2.80-3.41(m , 11H), 1.69-1.80 (m, 5H), 1.08 (d, 3H) ppm.

Example 8: (N-1-(8-((4-(morpholine-4-carbonyl)phenyl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl)pyrrole Preparation of Alk-3-yl)acrylamide (EXP8)

Referring to the method of Example 1, N-1-(8-((4-(morpholine-4-carbonyl)phenyl)amino)-1-oxo-1,2-dihydroisoquinoline-6 was prepared -yl)pyrrolidin-3-yl)acrylamide (EXP8) 71 mg, yield: 25%.

¹ H-NMR (DMSO-d6, 400MHz): δ12.23(br,1H), 11.87(br,1H), 8.32(br,1H), 7.68-7.70(m,5H), 6.45-6.90(m, 2H), 6.10-6.19 (m, 2H), 5.74 (d, 1H), 3.50-3.61 (m, 4H), 3.08-3.16 (m, 4H), 1.68-1.90 (m, 2H) ppm.

Example 9: N-(1-(1-oxo-8-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-1,2-dihydroisoquinoline Preparation of -6-yl)pyrrolidin-3-yl)acrylamide (EXP9)

Referring to the method of Example 1, N-(1-(1-oxo-8-((4-(2-(pyrrolidin-1-yl)ethyl)phenyl)amino)-1,2- Dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP9) 34 mg, yield: 32%. MS(ESI) m/z: [M+H] ⁺ =472.2. ¹ H-NMR (DMSO-d6, 400MHz): δ12.31(br,1H), 11.91(br,1H), 8.34(br,1H), 8.71(m,1H), 7.38(d,2H), 7.05 (d,2H),6.41-6.47(m,3H),6.10-6.19(m,2H),5.74(d,1H),3.71(m,1H),3.05-3.44(m,4H),2.74(t , 2H), 2.52-2.5 5 (m, 6H), 1.67-1.91 (m, 6H) ppm.

Example 10: N-(1-(8-((2-Methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-1-oxo-1,2-di Preparation of Hydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP10)

Referring to the method of Example 1, N-(1-(8-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-1-oxo- 1,2-Dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP10) 23 mg, yield: 19%. MS(ESI) m/z: [M+H] ⁺ =503.5. ¹ H-NMR (DMSO-d6, 400MHz): δ11.98(br,1H), 9.98(br,1H), 8.36(m,1H), 7.75(d,1H), 7.69(d,1H), 6.44 -6.48(m,3H),6.37(s,1H),5.75(m,1H),3.88(s,3H),3.72(m,1H),3.05-3.44(m,4H),2.36(t,4H) ), 2.18(s, 3H), 1.67- 1.90(m, 2H) ppm.

Example 11: (E)-3-(Dimethylamino)-N-(1-(8-((2-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino) - Preparation of 1-oxo-1,2-dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP11)

Referring to the method of Example 1, (E)-3-(dimethylamino)-N-(1-(8-((2-fluoro-4-(4-methylpiperazin-1-yl)) was prepared Phenyl)amino)-1-oxo-1,2-dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP11) 83 mg, yield: 47%. MS(ESI) m/z: [M+H] ⁺ =534.5. ¹ H-NMR (DMSO-d6, 400MHz): δ11.95(br,1H), 10.02(br,1H), 8.93(d,1H), 8.33(s,1H), 7.73(d,1H), 6.98 (d,1H), 6.82(d,1H), 6.53(m,1H), 6.44(m,2H), 6.21(d,1H), 5.25(m,1H), 3.73(m,1H), 3.15- 3.44(m, 7H), 3.06(m, 1H), 2.86(s, 6H), 2.38(t, 6H), 2.18(s, 3H), 1.65-1.90(m, 2H) ppm.

Example 12: N-(1-(8-((6-((4-methylpiperazin-1-yl)methyl)pyridin-3-yl)amino)-1-oxo-1,2- Preparation of Dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP12)

Referring to the method of Example 1, N-(1-(8-((6-((4-methylpiperazin-1-yl)methyl)pyridin-3-yl)amino)-1-oxo was prepared -1,2-Dihydroisoquinolin-6-yl)pyrrolidin-3-yl)acrylamide (EXP12) 78 mg, yield: 20%. MS(ESI) m/z: [M+H] ⁺ =488.2. ¹ H-NMR (DMSO-d6, 400MHz): δ11.98(br,1H), 9.25(s,1H), 8.33(s,1H), 7.70(m,2H), 7.45(d,1H), 7.19 (d,1H),6.45-6.52(m,3H),6.10-6.22(m,2H),5.75(m,1H),3.98(s,2H),3.59(m,1H),3.05-3.45(m , 4H), 2.45(t, 4H), 2.35(t, 4H), 2.13(s, 3H), 1.71-1.87(m, 2H) ppm.

Example 13: In vitro biochemical level inhibition of protein kinase activity assay

Materials and methods: BTK kinase, from Invitrogen; HTRF KinEASE; TK kit (Cisbio); 384-well plate (Greiner); ATP (sigma), MgCl ₂ (sigma); PHERAstar FS multifunctional microplate reader ( BMG company); low speed centrifuge (StaiteXiangyi company); incubator (Binder company). The selected positive drug is the Reference compound with the following structure:

Compound dissolution and storage: depending on the solubility, the test compound was prepared into a mother solution of 0.5-10 mmol/L with DMSO, and stored at -20 °C after packaging;

Preparation of compound working solution: before the test, take out the aliquoted compound from the refrigerator and dilute it to 50× the desired concentration with pure DMSO; then dilute the compound to 4× the desired concentration with deionized water;

1.33×Enzymatic buffer preparation: Dilute 5×Enzymatic buffer from HTRF kit) with deionized water to 1.33×, and add the corresponding components of 1.33× final concentration: 1.33mmol/L DTT and 1.33mmol/LMgCl2;

Preparation of kinase working solution: Dilute Btk to 2× the desired final concentration of 0.2ng/μL with 1.33×Enzymatic buffer;

Preparation of substrate working solution: Dilute substrate-biotin (from HTRF kit) and ATP (10mM) to a mixture of 4× desired final concentration with 1.33×Enzymatic buffer;

Preparation of detection working solution: 16.67 μmol/L Streptavidin-XL665 was diluted to 4× the desired final concentration with HTRF detection buffer, and then mixed with an equal volume of Antibody-Cryptate (both from HTRFkit).

Enzyme reaction steps: Add 4 μL of the kinase working solution to each well of a low-volume 384-well plate, and at the same time add 4 μL of 1.33×Enzymatic buffer as a negative control (Negative); add 2 μl of the compound working solution to the well, and add 2 μL of the working solution at the same time 8% DMSO aqueous solution was used as zero compound concentration control (i.e. positive control, Positive); incubate at 25°C (or 30°C) for 5-10 min; add 2 μL of substrate working solution to the well to start the enzyme reaction, and at 25°C (or 30°C) ℃) shaking reaction for 15-60min.

HTRF reagent detection steps: add 8 μL of detection working solution to the well to terminate the reaction; react at 25°C for 1 h;

HTRF signal reading: PHERAstar FS reading is used to detect the signal, and the instrument is set as follows:

Optic module

Integration delay(lag time)50μs

Integration time 400μs

Number of flashes 200

For raw data read per well, ratio = 665nm/620nm;

Calculation of Inhibition Rate:

Calculation of _IC50 value: take the logarithm of the compound concentration as the abscissa, and the inhibition rate as the ordinate. In GraphPad Prism 5, fit a nonlinear curve: log(inhibitor) vs.response--Variable slope, and calculate the enzyme activity inhibition The concentration of the test compound at 50% is the IC ₅₀ .

Experimental results: BTK kinase activity half inhibitory concentration (IC ₅₀ , nM)

The present invention provides the half inhibitory concentration (IC ₅₀ ) of the compound represented by formula I on BTK kinase activity

Reference compound EXP 1 EXP2 EXP3 EXP4 EXP5 EXP6 25.6nM 3.5nM 8.8nM 6.3nM 3.2nM 0.8nM 0.9nM EXP7 EXP8 EXP9 EXP10 EXP11 EXP12 2.4nM 0.5nM 8.2nM 2.6nM 1.5nM 5.8nM

Example: 14: In vitro cell level inhibition of protein kinase activity assay

Materials and methods: Fluo-4Direct ^™ Calcium Assay Kits, Invitrogen; RPMI1640 medium: GIBCO; 96-well black plate: CORNING; PHERAstar FS multifunctional microplate reader (BMG); low-speed centrifuge (StaiteXiangyi). The selected positive drug was RN486.

Cell Treatment: Wash cells with serum-free medium to remove serum.

Dye Preparation: The 2x dye was then diluted to 1x in serum-free medium.

Cell resuspension: Resuspend washed cells with 1X dye prepared above.

Cell seeding: 200,000/well, 40 μl/well, 96-well plate, black-walled plate required.

Dye incubation: put in an incubator and incubate for 40min.

Dosing: Then add a series of prepared compounds, 10 μl/well, and continue to act for 20 min.

Equilibration at room temperature: Take out the test plate and equilibrate at room temperature for 5 minutes.

Determination of baseline: Before adding agonist, use PHERStar microplate reader to detect the baseline.

Add agonist: Add IgM to a final concentration of 10 μg/ml, 10 μl/well.

Determination: After adding agonist, use PHERStar microplate reader for detection immediately, every 10s, for a total of 8min.

Data processing: OD (highest value) - OD (baseline), then IC50 values were calculated using GraphPad Prism 5 software.

Experimental results: The median inhibitory concentration of BTK cell calcium flux (IC ₅₀ , nM)

The present invention provides the half inhibitory concentration (IC ₅₀ ) of the compound of formula I on BTK cell calcium flux

RN486 EXP1 EXP2 EXP3 EXP4 EXP5 EXP6 45.7nM 3.2nM 3.5nM 10.2nM 8.9nM 5.2nM 6.5nM EXP7 EXP8 EXP9 EXP10 EXP11 EXP12 Reference compound 12.6nM 1.2nM 5.3nM 11.5nM 20.3nM 13.9nM 185nM

Example 15: Pharmacodynamic study on rat collagen-induced arthritis (rCIA) model

The compounds of the present invention inhibit the effect of rheumatoid arthritis. DBA/1J mice were selected, and 50ug of bovine type II collagen was completely emulsified with equal volume of complete Freund's adjuvant (CFA) and injected subcutaneously. After 21 days, 50ug of the same antigen and incomplete Freund's adjuvant (IFA) were fully emulsified, and then boosted 1 time. Observed and recorded from the 45th day. 1-4 scoring method: 1 point, normal; 2 points, 1 Joint swelling; 3 points, more than 1 joint swelling, but not all joints; 4 points, severe swelling or rigidity of the entire paw. The scores of each paw are added to obtain the total score of joint inflammation in the mouse. The total joint score Mice with more than 1 were successfully established. After the successful establishment of the mouse rheumatoid arthritis model, the compound of the present invention was used to give the mice intragastric administration, and the joint inflammation of the mice was scored after 2 weeks of administration. It has obvious therapeutic effect on rheumatoid arthritis in mice.

Compound group Arthritis Score Control 1 saline control group 4 Cyclosporine control group 2.4 EXP8 0.8 Reference compound 2.8

The foregoing invention has been described in some detail by way of illustration and example for the purposes of illustration and understanding. It will be apparent to those skilled in the art that changes and modifications may be practiced within the scope of the appended claims. Accordingly, it should be understood that the foregoing description is intended to be illustrative and not restrictive. The scope of the invention should, therefore, be determined not with reference to the above description, but should instead be determined with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

Claims (9)

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

wherein,

x, Y, Z, W is independently selected from CH or N;

ring A is C₆-C₁₀Aryl radical, C₆-C₁₀A heteroaryl group;

ring B is C₃-C₆Cycloalkyl radical, C₃-C₆A heterocycloalkyl group;

R₁、R₂selected from H, halogen, cyano, C₁-C₄Alkyl radical, C₁-C₃Alkoxy radical, C₃-C₆Cycloalkyl radical, C₃-C₆Heterocycloalkyl, said alkyl, alkoxy, cycloalkyl, heterocycloalkyl being optionally substituted by one or more halogens, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halo C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₃-C₆Heterocycloalkyl substitution;

R₃is selected from

2. The compound of the formula (I) or the pharmaceutically acceptable salt thereof according to claim 1, wherein,

z is selected from CH, W is selected from N or CH;

z is selected from N, and W is selected from CH.

3. The compound of claim 1, wherein ring A is a benzene ring or a pyridine ring, or a pharmaceutically acceptable salt thereof.

4. The compound of claim 1, having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein ring B is cyclohexane, cyclopentane, piperidine, morpholine, pyrrolidine.

5. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein at least one compound of formula (II),

wherein,

z is selected from CH, W is selected from N or CH; or Z is selected from N and W is selected from CH;

ring A is benzene ring or pyridine ring;

ring B is cyclohexane, cyclopentane, piperidine, morpholine, pyrrolidine;

R₁、R₂selected from H, halogen, cyano, C₁-C₄Alkyl radical, C₁-C₃Alkoxy radical, C₃-C₆Cycloalkyl radical, C₃-C₆Heterocycloalkyl, said alkyl, alkoxy, cycloalkyl, heterocycloalkyl being optionally substituted by one or more halogens, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halo C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₃-C₆Heterocycloalkyl substitution;

R₃is selected from

6. The compound according to claim 5, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

7. a pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically or acceptable excipient or diluent.

8. Use of a compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, for the manufacture of a BTK inhibitor.

9. Use of a compound according to claim 8, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of an inflammatory and/or autoimmune condition, wherein the inflammatory condition is rheumatoid arthritis.