HK40056552B

HK40056552B - Prophylactic and/or therapeutic agent for immune diseases

Info

Publication number: HK40056552B
Application number: HK42022046508.2A
Authority: HK
Inventors: 细井文仁; 仲地义宪; 梶原大辅
Original assignee: 大鹏药品工业株式会社
Priority date: 2015-01-30
Filing date: 2022-01-18
Publication date: 2025-05-02

Description

Prophylactic agents and/or therapeutic agents for immune diseases

(This application is a divisional application of application 201680007390.2, filed on January 29, 2016, entitled "Preventive and/or Therapeutic Agents for Immune Diseases")

Technical Field

This invention relates to preventive and/or therapeutic agents for immune diseases, particularly allergic diseases and autoimmune diseases, comprising novel fused pyrimidine compounds or their salts having Bruton’s tyrosine kinase (BTK) inhibitory activity as active ingredients.

Background Technology

It is well known that various protein kinases exist in living organisms and participate in a wide range of functional regulation. Bruton's tyrosine protein kinase (BTK) is a protein kinase belonging to the Tec kinase family. It is expressed in bone marrow cells such as B cells, monocytes/macrophages, neutrophils, mast cells, and osteoclasts, and participates in the functional control of these cells (Non-Patent Literature 1, 2). BTK is located downstream of immune receptor signaling such as B cell receptors (BCR) or Fc receptors (FcR), and participates in proliferation, survival, differentiation, and activation in B cells. In monocytes/macrophages or mast cells, it participates in the expression control of inflammatory cytokines (such as tumor necrosis factor-α or interleukin-1β) and chemical mediators (such as histamine or leukotrienes) (Non-Patent Literature 3). Inhibitors that can control BTK activity can be considered useful as therapeutic agents for diseases associated with abnormally hyperactive BTK signaling pathways (such as cancer, allergic diseases, and autoimmune diseases).

In recent years, it has been recognized that, in addition to B cells involved in antibody production, various cells expressing the Fc receptor (FcR) family or related molecules, such as monocytes/macrophages, neutrophils, mast cells, and osteoclasts, are also closely related to the pathogenesis and progression of autoimmune diseases such as rheumatoid arthritis (Non-Patent Literature 4). Since BTK signaling is associated with the activation of these cells or their abnormally hyperactive function (Non-Patent Literatures 2, 3), compounds with BTK inhibitory activity are expected to have therapeutic effects against autoimmune diseases. Furthermore, since BTK is also associated with mast cell activation, compounds with BTK inhibitory activity are expected to have therapeutic effects against allergic diseases involving B cells or mast cells.

Currently known BTK inhibitors include PCI-32765 (Non-Patent Document 5) and compounds described in Patent Documents 1 and 2 (Patent Documents 1 and 2). PCI-32765 is known to be a compound that can inhibit BTK and EGFR and is useful as a therapeutic agent for immune diseases (such as rheumatoid arthritis) (Non-Patent Document 5).

On the other hand, EGFR is known to bind to epidermal growth factor (EGF) and other ligands, participating in the proliferation and survival of various cells (such as inhibiting apoptosis) (Non-Patent Literature 6). However, it is known that side effects such as skin disorders or gastrointestinal disorders commonly occur when using inhibitors targeting EGFR. These side effects are generally believed to be related to the inhibition of the wild-type EGFR signaling pathway (Non-Patent Literature 7). Therefore, there is a search for inhibitors with high usefulness and high BTK selectivity as preventive and/or therapeutic agents for immune diseases.

Existing technical documents

Patent documents

Patent Document 1: International Publication No. 2011/090760

Patent Document 2: International Publication No. 2009/158571

Non-patent literature

Non-patent literature 1: Schaeffer and Schwartzberg, Curr Op Imm, 2000, pp. 282-288

Non-patent literature 2: Schmidt U., et al., Int Arch Allergy Immunol, 134, 2004,

Non-patent literature 3: Ellmeier W., et al., FEBS Journal., 278, 2011

Non-patent literature 4: Rommel C., et al., Nature Reviews Immunology, 7, 2007

Non-patent literature 5: Honigberg LA., et al., Proc Natl Acad Sci USA, 107, 2010

Non-patent literature 6: Lacouture ME., Nature Reviews Cancer, 6, 2006

Non-patent literature 7: Keefe DM and Bateman EH. Nature Reviews Clinical Oncology. 9, 2012

Summary of the Invention

The problem that the invention aims to solve

If the inhibitory activity of BTK and EGFR differs significantly, it is expected that the aforementioned side effects can be reduced.

In other words, from the perspective of reducing side effects, there is a desire for preventive and/or therapeutic agents for immune diseases that have high inhibitory activity against BTK and low inhibitory activity against other kinases such as EGFR, and also have excellent efficacy.

Therefore, the objective of this invention is to provide a preventive and/or therapeutic agent for immune diseases that is more effective than existing BTK inhibitors and has fewer side effects, particularly for allergic diseases and autoimmune diseases.

Methods for solving problems

In order to solve the above-mentioned problems, the inventors of the present invention conducted in-depth research and found that the compound group represented by the following general formula (I) showed excellent inhibitory activity and selectivity for BTK, and was useful as a medicine for treating autoimmune diseases such as rheumatoid arthritis or systemic lupus erythematosus, or immune diseases such as allergic diseases such as atopic dermatitis, thus completing the present invention.

That is, the present invention provides a preventive and/or therapeutic agent for immune diseases that uses a compound or salt thereof represented by the following general formula (I) as an active ingredient.

(In the formula, X represents a nitrogen-containing C3-C10 heterocyclic alkyl group that may have substituents;

Y represents -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z independently represent N or CH, respectively;

n represents an integer from 0 to 2;

R ₁ represents an amino group that can have substituents;

_R2 and _R3 may be the same or different, representing hydrogen atom, halogen atom, C1-C6 alkyl group that may have substituents, C1-C6 alkoxy group that may have substituents, C3-C7 cycloalkyl group that may have substituents, C6-C14 aromatic hydrocarbon group that may have substituents, 4-10 member monocyclic or polycyclic unsaturated heterocyclic group containing 1 to 3 heteroatoms of the same or different kinds selected from nitrogen, oxygen and sulfur atoms, or cyano group;

_R4 , _R5 , _R6 , and _R7 may be the same or different, representing a hydrogen atom or a C1-C6 alkyl group that may have substituents.

The present invention also provides compounds of the above general formula (I) or salts thereof for the prevention and/or treatment of immune diseases.

The present invention also provides the use of compounds of the above general formula (I) or salts thereof for the manufacture of preventive and/or therapeutic agents for immune diseases.

In addition, the present invention also provides a method for the prevention and/or treatment of immune diseases characterized by administering a compound of the above general formula (I) or a salt thereof.

As described above, PCI-32765 is known as a BTK inhibitor characterized by having a phenoxyphenyl group, while the compounds of the present invention are characterized by having a benzoxazolyl or oxazolopyridyl group, which is a significant difference between the two. Furthermore, as described later, the compounds of the present invention possess higher BTK selectivity compared to PCI-32765 (Comparative Compound 1).

Furthermore, the compounds described in Patent Documents 1 and 2 do not possess the benzoxazolyl or oxazolopyridyl groups characteristic of the compounds of the present invention, and their structures are significantly different.

In addition, the compound disclosed in International Publication No. 2007/067781 is known.

However, the compounds disclosed herein are inhibitors of aurora kinases, and no specific compounds containing benzoxazolyl or oxazolopyridyl groups are disclosed. Furthermore, there is no record of whether they possess BTK inhibitory activity, nor of their usefulness as a preventative and/or therapeutic agent for immune diseases.

Invention Effects

According to the present invention, it is possible to provide a preventive and/or therapeutic agent for immune diseases, particularly allergic diseases and autoimmune diseases, which is useful as a BTK inhibitor and contains the novel compound of the above general formula (I) or its salt as an active ingredient.

It is known that the compounds or salts thereof of the present invention possess excellent BTK inhibitory activity and exhibit superior efficacy in immune disease models. Furthermore, the compounds or salts thereof of the present invention can selectively and potently inhibit BTK compared to EGFR, thus reducing side effects and potentially improving safety. Therefore, the compounds or salts thereof of the present invention are useful as preventative and/or therapeutic agents for immune diseases, particularly allergic diseases or autoimmune diseases.

Attached Figure Description

Figure 1 shows the effect on the mouse collagen-induced arthritis model (preventive effect).

Figure 2 shows the effect on the mouse collagen-induced arthritis model (therapeutic effect).

Figure 3 shows the effect on the mouse collagen-induced arthritis model (therapeutic effect).

Figure 4 illustrates the effect of guinea pig antigen-induced rhinitis model.

Figure 5 shows the effect on a mouse model of systemic lupus erythematosus.

Detailed Implementation

The compound represented by formula (I) of the present invention is a compound having a 1H-pyrazolo[3,4-d]pyrimidine skeleton or a 7H-pyrrolo[2,3-d]pyrimidine skeleton, with benzoxazolyl or oxazolopyridyl as a substituent via an amide bond, and is a novel compound not described in any of the above-mentioned prior art documents.

In this application specification, the term "substituent" may include, for example, halogen atom, hydroxyl group, cyano group, nitro group, alkyl group, haloalkyl group, cycloalkyl group, cycloalkyl-alkyl group, aralkyl group, alkenyl group, alkoxy group, haloalkoxy group, cycloalkoxy group, cycloalkyl-alkoxy group, aralkyloxy group, alkylthio group, cycloalkyl-alkylthio group, amino group, mono- or dialkylamino group, cycloalkyl-alkylamino group, acyl group, acyloxy group, oxy group, carboxyl group, alkoxycarbonyl group, aralkyloxycarbonyl group, carbamoyl group, saturated or unsaturated heterocyclic group, aromatic hydrocarbon group, saturated heterocyclic group, etc. When the above substituents are present, the number of them is typically 1, 2 or 3.

In this application specification, fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms may be listed as "halogen atoms".

In this application specification, "alkyl" can be either straight-chain or branched, such as C1-C6 alkyl groups including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, and hexyl.

In this application specification, "halogenated alkyl" refers to a straight-chain or branched alkyl group (halogenated C1-C6 alkyl) having 1 to 13 halogen atoms and 1 to 6 carbon atoms. Examples of halogenated C1-C6 alkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, fluoroethyl, 1,1,1-trifluoroethyl, monofluoropropyl, perfluoropropyl, and perfluoroisopropyl, with halogenated C1-C4 alkyl groups being preferred.

In this application specification, specific examples of "cycloalkyl" include C3-C7 cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. In this application specification, "cycloalkylene" refers to a divalent cycloalkyl group.

In this application specification, "cycloalkyl-alkyl" can be used to refer to C3-C7 cycloalkyl-substituted C1-C4 alkyl groups such as cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and cycloheptylmethyl.

In this application specification, the term "aryl group" can be exemplified by C7-C13 aryl groups such as benzyl, phenethyl, naphthylmethyl, and fluorenemethyl.

In this application specification, "alkenyl" can be any of the following: linear, branched, or cyclic. It refers to an unsaturated hydrocarbon group having at least one double bond. Examples include vinyl, allyl, 1-propenyl, 2-methyl-2-propenyl, isopropenyl, 1-, 2-, or 3-butenyl, 2-, 3-, or 4-pentenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 5-hexenyl, 1-cyclopentenyl, 1-cyclohexenyl, 3-methyl-3-butenyl, and other C2-C6 alkenyl groups.

In this application specification, "alkynyl" can be any of the following: straight-chain, branched, or cyclic. It refers to an unsaturated hydrocarbon group having at least one triple bond, such as ethynyl, 1- or 2-propynyl, 1-, 2- or 3-butynyl, 1-methyl-2-propynyl, and other C2-C6 alkynyl groups.

In this application specification, "alkoxy" can be either linear or branched, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy, isopentoxy, and hexoxy, which are C1-C6 alkoxy groups.

In this application specification, "haloalkoxy" refers to a straight-chain or branched alkoxy group (haloC1-C6 alkoxy) having 1 to 13 halogen atoms and 1 to 6 carbon atoms. Examples include fluoromethoxy, difluoromethoxy, trifluoromethoxy, trichloromethoxy, fluoroethoxy, 1,1,1-trifluoroethoxy, monofluoropropoxy, perfluoropropoxy, perfluoroisopropoxy, and other haloC1-C6 alkoxy groups. HaloC1-C4 alkoxy groups are preferred.

In this application specification, specific examples of "cycloalkoxy" include C3-C7 cycloalkoxys such as cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexoxy, and cycloheptoxy.

In this application specification, "cycloalkyl-alkoxy" can be used to list C3-C7 cycloalkyl-substituted C1-C4 alkoxy groups such as cyclopropylmethoxy, cyclobutylmethoxy, cyclopentylmethoxy, cyclohexylmethoxy, and cycloheptylmethoxy.

In this application specification, the term "arylalkoxy" can be used to list C7-C13 arylalkoxys such as benzyloxy, phenylethoxy, naphthylmethoxy, and fluorenemethyloxy.

In this application specification, "alkathioyl" can be either linear or branched, such as methylthioyl, ethylthioyl, n-propylthioyl, isopropylthioyl, n-butylthioyl, isobutylthioyl, tert-butylthioyl, n-pentylthioyl, isopentylthioyl, hexylthioyl, and other C1-C6 alkathioyl groups.

In this application specification, "cycloalkyl-alkathio" can be used to refer to C3-C7 cycloalkyl-substituted C1-C4 alkathio groups such as cyclopropylmethylthio, cyclobutylmethylthio, cyclopentylmethylthio, cyclohexylmethylthio, and cycloheptylmethylthio.

In this application specification, "monoalkylamino" can be used to list amino groups that are monosubstituted with a straight-chain or branched C1-C6 alkyl group, such as methylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, tert-butylamino, n-pentylamino, isopentylamino, and hexylamino.

In this application specification, "dialkylamino" can refer to dimethylamino, diethylamino, di(n-propyl)amino, diisopropylamino, di(n-butyl)amino, diisobutylamino, di(tert-butyl)amino, di(n-pentyl)amino, diisopentylamino, dihexylamino, and other amino groups that are C1-C6 alkyl disubstituted in a straight-chain or branched manner.

In this application specification, "cycloalkyl-alkylamino" can be used to refer to C3-C7 cycloalkyl-substituted C1-C4 alkylamino compounds such as cyclopropylmethylamino, cyclobutylmethylamino, cyclopentylmethylamino, cyclohexylmethylamino, and cycloheptylmethylamino.

In this application specification, "acyl" refers to alkyl carbonyl or aryl carbonyl.

In this application specification, the term "alkyl carbonyl" can be used to list linear or branched (C1-C6 alkyl) carbonyl groups such as methyl carbonyl, ethyl carbonyl, n-propyl carbonyl, isopropyl carbonyl, n-butyl carbonyl, isobutyl carbonyl, tert-butyl carbonyl, n-pentyl carbonyl, isopentyl carbonyl, and hexyl carbonyl.

In this application specification, the term "aryl carbonyl" can be used to list phenyl carbonyl, naphthyl carbonyl, fluorenyl carbonyl, anthracene carbonyl, biphenyl carbonyl, tetrahydronaphthyl carbonyl, benzodihydropyranyl carbonyl, 2,3-dihydro-1,4-dioxananaphthyl carbonyl, indanyl carbonyl, and phenanthrene carbonyl (C6-C13 aryl) carbonyl groups.

In this application specification, "acyloxy" refers to alkyl carbonyloxy or aryl carbonyloxy.

In this application specification, the term "alkyl carbonyloxy" can be used to list linear or branched (C1-C6 alkyl) carbonyloxy compounds such as methyl carbonyloxy, ethyl carbonyloxy, n-propyl carbonyloxy, isopropyl carbonyloxy, n-butyl carbonyloxy, isobutyl carbonyloxy, tert-butyl carbonyloxy, n-pentyl carbonyloxy, isopentyl carbonyloxy, and hexyl carbonyloxy.

In this application specification, the term "aryl carbonyloxy" can be used to list phenyl carbonyloxy, naphthyl carbonyloxy, fluorenyl carbonyloxy, anthracene carbonyloxy, biphenyl carbonyloxy, tetrahydronaphthyl carbonyloxy, benzodihydropyranyl carbonyloxy, 2,3-dihydro-1,4-dioxananaphthyl carbonyloxy, dihydroindene carbonyloxy, and phenanthrene carbonyloxy, etc. (C6-C13 aryl) carbonyloxy.

In this application specification, "alkoxycarbonyl" can be either linear or branched, such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, pentoxycarbonyl, isopentoxycarbonyl, and hexoxycarbonyl (C1-C6 alkoxy) carbonyl groups.

In this application specification, "araneoxycarbonyl" can be used to list benzyloxycarbonyl, phenylethoxycarbonyl, naphthylmethoxycarbonyl, fluorenemethoxycarbonyl, and other (C7-C13 aranealkyl) oxycarbonyl groups.

In this application specification, "saturated heterocyclic group" refers to a saturated heterocyclic group having a heteroatom selected from nitrogen, oxygen and sulfur atoms, specifically including morpholino, 1-pyrrolidinyl, piperidinyl, piperazine, 4-methyl-1-piperazine, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, thiazolyl, and oxazolyl.

In this application specification, "unsaturated heterocyclic group" refers to a monocyclic or polycyclic heterocyclic group having a heteroatom selected from nitrogen, oxygen, and sulfur atoms, and being completely or partially unsaturated. Specific examples include imidazole, thiophene, furanyl, pyrrole, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolyl, thiadiazolyl, pyrazolyl, triazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridyl, indole, isindole, inzolyl, triazolopyridinyl, benzimidazole, benzoxazolyl, benzothiazolyl, benzothiophene, benzofuranyl, purine, quinolinyl, isoquinolinyl, quinazolinyl, quinoxolinyl, methylenedioxyphenyl, ethylenedioxyphenyl, and dihydrobenzofuranyl.

In this application specification, the term "aromatic hydrocarbon group" can be used to list C6-C14 aromatic hydrocarbon groups such as phenyl, tolyl, xylyl, naphthyl, anthracene, phenanthryl, fluorenyl, and tetrahydronaphthyl.

In this application specification, "saturated heterocyclic oxy group" refers to an oxy group bonded with a saturated heterocyclic group having a heteroatom selected from nitrogen, oxygen and sulfur atoms. Specific examples include morpholinyloxy, 1-pyrrolidinyloxy, piperidinyloxy, piperazinyloxy, 4-methyl-1-piperazinyloxy, tetrahydrofuranyloxy, tetrahydropyranyloxy, tetrahydrothiophenyloxy, thiazolyloxy, and oxazolidinyloxy.

Furthermore, in the description of substituents in this specification, "CA-CB" indicates a substituent with a carbon number of A to B. For example, "C1-C6 alkyl" indicates an alkyl group with 1 to 6 carbon atoms, and "C6-C14 aromatic hydrocarbon oxy" indicates an oxygen group bonded to an aromatic hydrocarbon group with 6 to 14 carbon atoms. Additionally, "A-B member" indicates the number of atoms constituting the ring (ring element number) of A to B. For example, "4-10 member saturated heterocyclic group" indicates a saturated heterocyclic group with a ring element number of 4 to 10.

In general formula (I), X represents a divalent heterocyclic alkyl group (nitrogen-containing C3-C10 heterocyclic alkyl group) with 3 to 10 carbon atoms, which may have substituents, contains at least one nitrogen atom in the ring, and contains 0 to 2 heteroatoms of the same or different kinds selected from oxygen or sulfur atoms in the ring. Examples include pyridylene, pyrrolidinylene, piperidinylene, piperazineyl, morpholinyl, octahydroquinolinyl, octahydroindolyl, etc.

Preferably, it is a nitrogen-containing C3-C5 heterocyclic alkyl group with 3 to 5 carbon atoms and one nitrogen atom in the ring, which may have substituents; more preferably, it is acrylonitrile, pyrrolidine, or piperidinyl; and even more preferably, it is 1,3-acrylonitrile, 1,3-pyrrolidine, or 1,3-piperidine.

As substituents on these heterocyclic alkyl groups, the above-mentioned substituents can be listed, preferably unsubstituted.

The nitrogen atom of the nitrogen-containing C3-C10 heterocyclic alkyl group shown in X is preferably bonded to the carbonyl group of -COY in general formula (I).

Furthermore, the nitrogen atom of the nitrogen-containing C3-C5 heterocyclic alkyl group shown in X is preferably bonded to the carbonyl group of -COY in general formula (I).

In general formula (I), Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 .

In general formula (I), W and Z are independently N or CH, preferably Z is N and W is N, or Z is CH and W is N or CH.

In general formula (I), n is preferably 0.

In general formula (I), the "substituent" in "amino group that may have substituents" as shown in R ₁ can be the substituents mentioned above, preferably unsubstituents.

The "amino group that may have substituents" indicated by R ₁ is preferably an amino group.

In general formula (I), the "halogen atom" represented by _R2 or _R3 is preferably a fluorine atom, a chlorine atom, or a bromine atom.

In general formula (I), "C1-C6 alkyl" in "may have substituents" represented by _R2 or _R3 is preferably C1-C4 alkyl, more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, and even more preferably methyl or ethyl.

The "substituent" in "C1-C6 alkyl group that may have substituents" as shown in _R2 or _R3 is preferably unsubstituted, a halogen atom, or a C1-C4 alkoxy group, more preferably unsubstituted, a fluorine atom, or a methoxy group. When substituents are present, the number of substituents is not particularly limited; 1 to 3 substituents are preferred when the substituent is a halogen atom, and 1 substituent is preferred when the substituent is a C1-C4 alkoxy group.

The "C1-C6 alkyl group that may have substituents" indicated by _R2 or _R3 is preferably a C1-C6 alkyl group, a halo-C1-C6 alkyl group, or a C1-C4 alkoxy-substituted C1-C6 alkyl group, more preferably a C1-C4 alkyl group, a halo-C1-C4 alkyl group, or a C1-C4 alkoxy-substituted C1-C4 alkyl group, further preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, trifluoromethyl, trichloromethyl, methoxyethyl, or ethoxyethyl, and even more preferably methyl, trifluoromethyl, or methoxyethyl.

In general formula (I), "C1-C6 alkoxy" which is represented as _R2 or _R3 as "C1-C6 alkoxy that may have substituents" is preferably "C1-C4 alkoxy", more preferably methoxy, ethoxy, isopropoxy or n-butoxy, and even more preferably methoxy.

As "substituent" in "C1-C6 alkoxy group that may have substituents" as shown in _R2 or _R3 , substituents as described above can be listed, preferably unsubstituented.

The "C1-C6 alkoxy group that may have substituents" represented by _R2 or _R3 is preferably C1-C6 alkoxy, more preferably C1-C4 alkoxy, further preferably methoxy, ethoxy, isopropoxy or n-butoxy, and even more preferably methoxy.

In general formula (I), "C3-C7 cycloalkyl" as _R2 or _R3 , which may have substituents, is preferably C3-C6 cycloalkyl, more preferably cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

As "substituent" in "C3-C7 cycloalkyl group that may have substituents" as shown in _R2 or _R3 , the substituents described above can be exemplified, and preferably unsubstituented.

The "C3-C7 cycloalkyl group that may have substituents" represented by _R2 or _R3 is preferably a C3-C6 cycloalkyl group, more preferably a cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl group.

In general formula (I), the "C6-C14 aromatic hydrocarbon group" represented by _R2 or _R3 is preferably phenyl or naphthyl, and more preferably phenyl.

The "substituent" in "C6-C14 aromatic hydrocarbon group that may have substituents" shown in _R2 or _R3 is preferably unsubstituted or has a halogen atom, more preferably unsubstituted or has a chlorine atom or a fluorine atom. When substituents are present, the number of substituents is not particularly limited, but is preferably 1 to 3.

The "C6-C14 aromatic hydrocarbon group that may have substituents" indicated by _R2 or _R3 is preferably unsubstituted or can be substituted with halogen atoms of phenyl or naphthyl, more preferably phenyl, chlorophenyl, fluorophenyl, dichlorophenyl or trichlorophenyl, further preferably phenyl or chlorophenyl, and particularly preferably phenyl or 4-chlorophenyl.

In general formula (I), the "unsaturated heterocyclic group containing 1 to 3 heteroatoms of the same or different kinds selected from nitrogen, oxygen and sulfur atoms, which may have substituents" in _R2 or _R3, is preferably a 4- to 6-membered monocyclic or polycyclic unsaturated heterocyclic group containing 1 nitrogen atom, oxygen atom or sulfur atom, more preferably a 4- to 6-membered monocyclic unsaturated heterocyclic group containing 1 nitrogen atom, oxygen atom or sulfur atom, even more preferably a 4- to 6-membered monocyclic unsaturated heterocyclic group containing 1 sulfur atom, further preferably a thiophene group, and even more preferably a 2-thiophene group.

The "substituent" in "a 4- to 10-membered monocyclic or polycyclic unsaturated heterocyclic group containing 1 to 3 heteroatoms selected from the same or different types of nitrogen, oxygen and sulfur atoms" as shown in _R2 or _R3 can be exemplified by the above-mentioned substituent, preferably unsubstituted.

The “4- to 10-membered monocyclic or polycyclic unsaturated heterocyclic group containing 1 to 3 heteroatoms selected from the same or different types of nitrogen, oxygen and sulfur atoms” shown in _R2 or _R3 is preferably a 4- to 6-membered monocyclic unsaturated heterocyclic group containing 1 nitrogen atom, oxygen atom or sulfur atom, more preferably a 4- to 6-membered monocyclic unsaturated heterocyclic group containing 1 sulfur atom, further preferably a thiophene group, and even more preferably a 2-thiophene group.

In general formula (I), "C1-C6 alkyl" as "may have substituents" represented by _R4 , _R5 or _R6 is preferably C1-C4 alkyl, more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, and even more preferably methyl.

The "substituent" in "C1-C6 alkyl group that may have substituents" as shown in _R4 , _R5 or _R6 is preferably unsubstituted or substituted with an amino group having two C1-C4 alkyl groups (the C1-C4 alkyl groups can form a heterocyclic alkyl group with a 4- to 8-membered ring together with the nitrogen atom to which they are bonded), more preferably unsubstituted, dimethylamino, methylethylamino, diethylamino, methylisopropylamino, 1-piperidinyl or 1-pyrrolidinyl. When the "C1-C6 alkyl group that may have substituents" has substituents, the number of substituents is not particularly limited, but is preferably one.

The "C1-C6 alkyl group that may have substituents" represented by _R4 , _R5 or _R6 is preferably a C1-C4 alkyl group, or a C1-C4 alkyl group substituted with an amino group having two C1-C4 alkyl groups (the C1-C4 alkyl group can form a heterocyclic alkyl group with a nitrogen atom to which it is attached, together with the nitrogen atom to which it is attached), and more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, dimethylaminomethyl, methylethylaminomethyl, diethylaminomethyl, methylisopropylaminomethyl, dimethylaminoethyl, diethylaminoethyl, 1-piperidinylmethyl or 1-pyrrolidinylmethyl.

In general formula (I), "C1-C6 alkyl" in "may have substituents" shown in R ₇ is preferably C1-C4 alkyl, more preferably methyl, ethyl, n-propyl, isopropyl or n-butyl, and even more preferably methyl.

As for "substituent" in "C1-C6 alkyl group that may have substituents" as shown in R ₇ , the above-mentioned substituents can be exemplified, and preferably unsubstituented.

As indicated by R ₇ , "C1-C6 alkyl group which may have substituents" is preferably C1-C4 alkyl group, more preferably methyl, ethyl, n-propyl, isopropyl or n-butyl, and even more preferably methyl.

In general formula (I), the -C( _R4 )=C( _R5 )( _R6 ) or -C≡C- _R7 shown in Y is particularly preferred to be any one of the following groups:

In the compounds of the present invention represented by general formula (I), the preferred option is:

X is a nitrogen-containing C3-C10 heterocyclic alkyl group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

_R2 and _R3 may be the same or different, and may be a hydrogen atom, a halogen atom, a C1-C6 alkyl group that may have substituents, a C1-C6 alkoxy group that may have substituents, a C3-C7 cycloalkyl group that may have substituents, a C6-C14 aromatic hydrocarbon group that may have substituents, a 4-10 member monocyclic or polycyclic unsaturated heterocyclic group containing 1 to 3 heteroatoms selected from the same or different types of nitrogen, oxygen and sulfur atoms, or a cyano group;

_R4 , _R5 , _R6 , and _R7 may be the same or different, and may be hydrogen atoms or C1-C6 alkyl compounds or salts thereof that may have substituents.

In this case, the preferred form of the compound of the present invention represented by general formula (I) is:

X is a nitrogen-containing C3-C10 heterocyclic alkyl group (here, the nitrogen atom is bonded to the carbonyl group of the general formula (I) -COY);

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

More preferably, in the compounds of the present invention represented by general formula (I), the following are preferred:

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

In this case, the compound of the present invention represented by general formula (I) is more preferably:

X is acrylonitrile, pyrrolidinyl or piperidinyl (here, the nitrogen atom is bonded to the carbonyl group of -COY of general formula (I));

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a C1-C6 alkyl group, and the other is a hydrogen atom, a halogen atom, a C1-C6 alkyl group, a halo-C1-C6 alkyl group, a C1-C4 alkoxy-substituted C1-C6 alkyl group, a C1-C6 alkoxy group, a phenyl group that can be substituted with a halogen atom, a 4- to 6-membered monocyclic unsaturated heterocyclic group containing one sulfur atom, or a cyano group.

When Y is -C( _R4 )=C( _R5 )( _R6 ), _R4 , _R5 and _R6 may be the same or different, and may be a hydrogen atom, a C1-C6 alkyl group, or a C1-C6 alkyl group substituted with an amino group having two C1-C6 alkyl groups (C1-C6 alkyl groups can form heterocyclic alkyl groups with the nitrogen atoms they are bonded to); when Y is -C≡C- _R7 , _R7 may be a hydrogen atom or a C1-C6 alkyl group, a compound or a salt thereof.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a C1-C4 alkyl group, and the other is a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a halo-C1-C4 alkyl group, a C1-C4 alkoxy-substituted C1-C4 alkyl group, a C1-C4 alkoxy group, a phenyl group that can be substituted with a halogen atom, a 4- to 6-membered monocyclic unsaturated heterocyclic group containing one sulfur atom, or a cyano group.

When Y is -C( _R4 )=C( _R5 )( _R6 ), _R4 , _R5 and _R6 may be the same or different, and may be a hydrogen atom, a C1-C6 alkyl group, or a C1-C6 alkyl group substituted with an amino group having two C1-C6 alkyl groups (C1-C6 alkyl groups can form heterocyclic alkyl groups with the nitrogen atoms they are bonded to); when Y is -C≡C- _R7 , _R7 may be a hydrogen atom or a C1-C4 alkyl group, a compound or a salt thereof.

X is 1,3-adiazinyl, 1,3-pyrrolidinyl, or 1,3-piperidinyl (here, the nitrogen atom is bonded to the carbonyl group of -COY of general formula (I));

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

When Y is -C( _R4 )=C( _R5 )( _R6 ), _R4 , _R5 and _R6 may be the same or different, and may be a hydrogen atom, a C1-C6 alkyl group, or a C1-C6 alkyl group substituted with an amino group having two C1-C6 alkyl groups (C1-C6 alkyl groups can form heterocyclic alkyl groups with the nitrogen atoms they are bonded to); when Y is -C≡C- _R7 , _R7 may be an atom or a C1-C4 group, a compound or a salt thereof.

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a methyl group, and the other is a hydrogen atom, a halogen atom, a methyl group, a trifluoromethyl group, a methoxyethyl group, a methoxy group, a phenyl group, a 4-chlorophenyl group, a 2-thienyl group, or a cyano group;

When Y is -C( _R4 ) = C( _R5 )( _R6 ), _R4 , _R5 , and _R6 may be the same or different, and can be hydrogen atoms, methyl, dimethylaminomethyl, methylethylaminomethyl, diethylaminomethyl, methylisopropylaminomethyl, 1-piperidinylmethyl, or 1-pyrrolidinylmethyl; when Y is -C≡C- _R7 , _R7 is a methyl compound or its salt.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

(1) When Z is N and W is N.

X is 1,3-piperidinyl,

Y represents vinyl.

(2) When Z is CH and W is N.

X is 1,3-pyridinealkyl or 1,3-piperidineyl.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

When Y is -C( _R4 ) = C( _R5 )( _R6 ), _R4 , _R5 , and _R6 may be the same or different, and can be hydrogen atoms, methyl, dimethylaminomethyl, methylethylaminomethyl, diethylaminomethyl, methylisopropylaminomethyl, 1-piperidinylmethyl, or 1-pyrrolidinylmethyl; when Y is -C≡C-( _R7 ), _R7 is a methyl group.

(3) When Z is CH and W is CH.

X is 1,3-adiazine-1,3-pyrrolidine-1,3-diphenylene ...

Y is -C( _R4 ) = C( _R5 )( _R6 ).

_R4 , _R5 and _R6 may be the same or different, and may be hydrogen atoms, dimethylaminomethyl, methylethylaminomethyl, diethylaminomethyl, methylisopropylaminomethyl, 1-piperidinylmethyl or 1-pyrrolidinylmethyl;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a methyl group, and the other is a hydrogen atom, a halogen atom, a trifluoromethyl group, a methoxyethyl group, a phenyl group, a 2-thiophene group, or a cyano group, a compound or a salt thereof.

(1) When Z is N and W is N.

X is 1,3-piperidine (here, the nitrogen atom is bonded to the carbonyl group of the general formula (I) -COY),

Y represents vinyl.

(2) When Z is CH and W is N.

X is 1,3-pyridinealkyl or 1,3-piperidineyl (here, the nitrogen atom is bonded to the carbonyl group of the general formula (I) -COY),

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

(3) When Z is CH and W is CH.

X is 1,3-adiazine-1,3-pyrrolidine-1 (here, the nitrogen atom is bonded to the carbonyl group of -COY in general formula (I)).

Y is -C( _R4 ) = C( _R5 )( _R6 ).

n is 0;

_R1 is an amino group;

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom, and the other is a hydrogen atom, a halogen atom, or a cyano group, a compound or a salt thereof.

X is 1,3-piperidine (here, the nitrogen atom is bonded to the carbonyl group of -COY in general formula (I));

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom, and the other is a hydrogen atom, a halogen atom, a cyano group, a compound, or a salt thereof.

Of the compounds of the present invention represented by general formula (I), those of the following are particularly preferred:

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom, and the other is a hydrogen atom or a halogen atom, a compound or a salt thereof.

In this case, the compounds of the present invention represented by general formula (I) are particularly preferred to be:

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

As specific compounds of the present invention, compounds obtained in the examples described below may be listed, but the invention is not limited to these compounds.

As preferred compounds of the present invention, the following compounds may be listed:

(1) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 1)

(2) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-bromobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 2)

(3) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 3)

(4)(R)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－1－(1－methacryloylpiperidin-3－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－formamide (Example Compound 4)

(5) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 5)

(6) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-cyanobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 6)

(7) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-methoxybenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 7)

(8) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-(2-methoxyethyl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 8)

(9) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(Oxazo[4,5-b]pyridin-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 9)

(10) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(4-methylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 10)

(11) (R)-4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)-1-(1-methacryloylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 11)

(12) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-fluorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 12)

(13) (R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(Benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 13)

(14) (R, E)-4-amino-N-(benzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 14)

(15) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 15)

(16) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(ethyl(methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 16)

(17) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(diethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 17)

(18) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 18)

(19) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyrrolidine-1-yl)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 19)

(20) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(piperidin-1-yl)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 20)

(21) (R, E)-4-amino-N-(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 21)

(22) (R)－4－amino－N－(benzo[d]oxazol-2－yl)－1－(1－(but-2－ynyl)piperidin-3－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide (Example Compound 22)

(23)(R)－1－(1-acryloylpiperidin-3-yl)－4-amino-N－(5,6-dimethylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 23)

(24)(R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 24)

(25) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 25)

(26) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(3-methylbut-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 26)

(27) (R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N-(benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 27)

(28) (R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N－(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 28)

(29) (R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N-(5-methylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 29)

(30)(R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 30)

(31)(R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N－(5-(4-chlorophenyl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 31)

(32) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 32)

(33) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 33)

(34) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(diethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 34)

(35) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 35)

(36) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyrrolidine-1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 36)

(37) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 37)

(38)(R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N－(5-methoxybenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 38)

(39) (R)－1－(1-Acryloylpyrrolidine-3-yl)－4-amino-N－(5-cyanobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 39)

(40) (R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-(2-methoxyethyl)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 40)

(41) (R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 41)

(42) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 42)

(43) (R)－1－(1-acryloylpyrrolidine-3-yl)－4-amino-N－(5-(trifluoromethyl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 43)

(44) (R,E)-4-amino-N-(5-(trifluoromethyl)benzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 44)

(45) 1-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 45)

(46) 7-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 46)

(47)(E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－7－(1－(4－(dimethylamino)but-2－enoyl)acetidin-3－yl)7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide (Example compound 47)

(48) (R)-7-(1-Acryloylpyrrolidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example Compound 48)

(49) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 49)

(50) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 50)

(51) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(diethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 51)

(52) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 52)

(53) (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(pyrrolidine-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 53)

(54) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 54)

(55)(R)－7－(1－acryloylpyrrolidine-3－yl)－4－amino-N－(5－phenylbenzo[d]oxazol-2－yl)－7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide (Example compound 55)

(56) (R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 56)

(57) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 57)

(58) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(diethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 58)

(59) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 59)

(60) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(pyrrolidine-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 60)

(61) (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example compound 61)

The method for manufacturing the compound of the present invention will now be described.

The compound (I) of the present invention can be manufactured by, for example, the manufacturing method described below or the method shown in the examples. However, the manufacturing method of the compound (I) of the present invention is not limited to the reaction examples.

Manufacturing Method 1

[In the formula, Z, _R2 , and _R3 have the same meaning as above.]

(Step 1) This step is a process for synthesizing a benzoxazole compound of general formula (III) from aminophenol of general formula (II). The compound of general formula (II) can be manufactured using commercially available products or based on known methods.

Examples of cyano compounds used as reagents include cyanogen bromide, cyanogen chloride, cyanogen iodide, and 1,1-carboiminobis-1H-imidazole. 0.5 to 5 moles, preferably 0.9 to 1.5 moles, of the cyano compound are used relative to 1 mole of the compound shown in general formula (II). This cyano compound can be commercially available or manufactured using known methods. Any solvent that does not adversely affect the reaction can be used, such as alcohols (e.g., methanol, ethanol), hydrocarbons (e.g., benzene, toluene, xylene), halogenated hydrocarbons (e.g., dichloromethane, chloroform, 1,2-dichloroethane), nitriles (e.g., acetonitrile), ethers (e.g., dimethoxyethane, tetrahydrofuran), aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide), water, or mixtures thereof. The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours. The reaction temperature is 0–120℃, preferably 0–90℃.

The compound of general formula (III) obtained by such operation can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc., or delivered to subsequent processes without separation and purification.

Manufacturing Method 2

[In the formula, _L3 and _L4 represent the dereliction groups; _P1 represents the protecting group of the amino group contained in X; W, X, Y, Z, _R1 , _R2 , _R3 and n have the same meaning as above.]

(Step 2) This step is a step of manufacturing a compound of general formula (VII) using compounds of general formula (IV), and general formula (V) or general formula (VI). The compound of general formula (IV) can be manufactured using commercially available products or based on known methods.

When the compound shown in general formula (V) is used as an alkylating agent, the preparation can be carried out in the presence of a base. In general formula (V), _L4 can be, for example, a detached group such as a chlorine atom, a bromine atom, an iodine atom, a methanesulfonate ester, or a p-toluenesulfonate ester, and can be prepared using commercially available products or based on known methods. The amount of the compound shown in general formula (V) is 1 to 10 moles, preferably 1 to 5 moles, relative to 1 mole of the compound shown in general formula (IV).

Examples of bases include inorganic bases such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, cesium hydroxide, sodium hydride, and potassium hydride, as well as organic amines such as trimethylamine, triethylamine, tripropylamine, diisopropylethylamine, N-methylmorpholine, pyridine, 4-(N,N-dimethylamino)pyridine, lutidine, and collidine. The amount of base used is 1 to 100 moles relative to 1 mole of the compound shown in general formula (IV), preferably 2 to 10 moles.

As a solvent, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, tetrahydrofuran, 1,4-dioxane, N-methylpyrrolidone-2-one, acetonitrile, etc., can be used alone or in combination. The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours. The reaction temperature is 0°C to the boiling temperature of the solvent, preferably 0 to 100°C.

When using general formula (VI) as an alkylating agent, it can be manufactured using a photoelectrophoresis reaction. This step can be carried out based on commonly known methods (e.g., Chemical Reviews, Vol. 109, p. 2551, 2009), for example, in the presence of photoelectrophoresis agents (Mitsunobu Reagents) and phosphine reagents, in a solvent that does not adversely affect the reaction. In this step, generally 1 to 10 moles, preferably 1 to 5 moles, of the compound represented by general formula (VI) are used relative to 1 mole of the compound represented by general formula (IV).

Examples of photocontinuation reagents include diethyl azodicarbonate and diisopropyl azodicarbonate. The amount of photocontinuation reagent used is 1 to 10 moles, preferably 1 to 5 moles, relative to 1 mole of the compound represented by general formula (IV).

Phosphine reagents include, for example, triphenylphosphine and tributylphosphine. The amount of phosphine reagent used is 1 to 10 moles, preferably 1 to 5 moles, relative to 1 mole of the compound represented by general formula (IV).

The reaction solvent can be any substance that does not hinder the reaction and is not particularly limited. For example, toluene, benzene, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, or mixtures thereof are preferred.

The reaction temperature is typically -78 to 200°C, preferably 0 to 50°C. The reaction time is typically 5 minutes to 3 days, preferably 10 minutes to 10 hours.

The compound obtained in this way, represented by general formula (VII), can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc., or it can be delivered to subsequent processes without separation and purification.

(Step 3) This step is a step of producing the compound of general formula (VIII) by reacting the compound of general formula (VII) with, for example, a transition metal and a base as needed, in a solvent that does not adversely affect the reaction in a carbon monoxide atmosphere and in the presence of an alcohol.

In general formula (VII), the detaching group represented by L ₃ is a bromine atom or an iodine atom, and the compound can be manufactured using commercially available products or based on known methods.

In this process, the carbon monoxide pressure is typically 1 to 10 atmospheres, preferably 1 to 5 atmospheres. The amount of alcohol compound used is 1 to 10 moles, preferably 1 to 5 moles, relative to 1 mole of the compound represented by general formula (VII). Examples of alcohol compounds include methanol, ethanol, propanol, isopropanol, diethylaminoethanol, isobutanol, 4-(2-hydroxyethyl)morpholine, 3-morpholinopropanol, and diethylaminopropanol.

Transition metal catalysts that can be used in this process include, for example, palladium catalysts (e.g., palladium acetate, tris(dibenzylacetone)dipalladium, bis(triphenylphosphine)palladium(II) dichloride, 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride-dichloromethane coordination compounds, etc.), and ligands (e.g., triphenylphosphine, 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (xantphos), tri-tert-butylphosphine, etc.) may be added as needed. The amount of transition metal catalyst used varies depending on the type of catalyst, and is typically 0.0001 to 1 mole, preferably 0.001 to 0.5 moles, relative to 1 mole of the compound shown in general formula (VII). The amount of ligand used is typically 0.0001 to 4 moles, preferably 0.01 to 2 moles, relative to 1 mole of the compound shown in general formula (VII).

Furthermore, a base may be added to the above reaction as needed. Examples of bases include triethylamine, diisopropylethylamine, pyridine, dimethylpyridine, trimethylpyridine, 4-dimethylaminopyridine, N-methylmorpholine, potassium tert-butyrate, sodium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, potassium hexamethyldisilazane, butyllithium, and other organic bases, or sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, sodium hydride, and other inorganic bases. The amount of base used is typically 0.1 to 50 moles, preferably 1 to 20 moles, relative to 1 mole of the compound shown in general formula (VII).

The reaction solvent can be any solvent that does not impede the reaction and is not particularly limited. Examples include hydrocarbons (e.g., benzene, toluene, xylene, etc.), nitriles (e.g., acetonitrile, etc.), ethers (e.g., dimethoxyethane, tetrahydrofuran, 1,4-dioxane, etc.), alcohols (e.g., methanol, ethanol, etc.), aprotic polar solvents (e.g., dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, hexamethylphosphoramide, etc.), water, or mixtures thereof.

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

The reaction temperature is 0℃ to the boiling temperature of the solvent, preferably 0 to 150℃.

Following this reaction, a mixture of a carboxylic acid compound (VIII) and an ester body corresponding to the alcohol used is formed. Therefore, a hydrolysis reaction is performed to convert it into a compound represented by the general formula (VIII). Hydrolysis is carried out using a base, such as organic bases like diethylamine, diisopropylamine, potassium tert-butyrate, sodium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, potassium hexamethyldisilazane, and butyllithium, or inorganic bases like sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, and sodium hydroxide.

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

The compound obtained in this way, represented by general formula (VIII), can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc., or it can be delivered to subsequent processes without separation and purification.

(Step 4) This step is to use compounds of general formula (VIII) and general formula (III) to carry out an amidation reaction to produce compounds of general formula (IX).

As an amidating agent, in the presence of a suitable condensing agent or activator, 0.5 to 10 moles, preferably 1 to 3 moles, of the compound of general formula (III) are used relative to 1 mole of the compound of general formula (VIII).

The reaction solvent is not particularly limited as long as it does not hinder the reaction. Preferred solvents include isopropanol, tert-butanol, toluene, benzene, dichloromethane, chloroform, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, or mixtures thereof. The reaction temperature is typically -78 to 200°C, preferably 0 to 50°C. The reaction time is typically 5 minutes to 3 days, preferably 5 minutes to 10 hours.

As condensing agents and activators, examples include diphenyl azidophosphate, N,N'-dicyclohexylcarbodiimide, benzotriazol-1-yloxy-tris-dimethylaminophosphonium salt, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, 1-ethyl-3-(3-dimethylaminophenyl)carbodiimide, combinations of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 1-hydroxybenzotriazole, and 2-chloro... -1,3-Dimethylimidazolium chloride, O-(7-azabenzotriazo-1-yl)-N,N,N',N'-tetramethylhexauronium hexafluoro phosphate, 1,1-carbonyldiimidazole, N-hydroxysuccinimide, etc.

Furthermore, a base may be added to the above reaction as needed. Examples of bases include triethylamine, diisopropylethylamine, pyridine, dimethylpyridine, trimethylpyridine, 4-dimethylaminopyridine, potassium tert-butyrate, sodium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, potassium hexamethyldisilazane, diazabicycloundecene, diazabicyclononene, butyllithium, etc., and inorganic bases such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, sodium hydride, etc. The amount added is 1 to 100 moles, preferably 1 to 10 moles, relative to 1 mole of the compound shown in general formula (VIII).

The compound of general formula (IX) thus obtained can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc., or can be used to manufacture the compound (I) of the present invention without separation and purification.

Manufacturing method 3

[In the formula, _L3 represents the detachment basis; the meanings of W, X, Y, Z, _P1 , _R1 , _R2 , _R3 , and n are the same as above.]

(Step 5) This step is a step of producing the compound of general formula (IX) by reacting the compound of general formula (VII) with, for example, a transition metal and a base as required, in a solvent that does not adversely affect the reaction in the presence of compound (III) under a carbon monoxide atmosphere.

In general formula (VII), the detaching group shown as L ₃ is a bromine atom or an iodine atom, and the compound can be manufactured using commercially available products or based on known methods.

In this process, the pressure of carbon monoxide is 1 atmosphere to 10 atmospheres, preferably 1 atmosphere to 5 atmospheres.

Transition metal catalysts that can be used in this process include, for example, palladium catalysts (e.g., palladium acetate, tris(dibenzylacetone)dipalladium, bis(triphenylphosphine)palladium(II) dichloride, 1,1'-bis(diphenylphosphine)ferrocene-palladium(II) dichloride-dichloromethane coordination compounds, etc.), and ligands (e.g., triphenylphosphine, 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene, tri-tert-butylphosphine, etc.) can be added as needed. The amount of transition metal catalyst used varies depending on the type of catalyst, and is typically 0.0001 to 1 mole, preferably 0.001 to 0.5 moles, relative to 1 mole of the compound shown in general formula (IX). The amount of ligand used is typically 0.0001 to 4 moles, preferably 0.01 to 2 moles, relative to 1 mole of the compound shown in general formula (VII).

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

Manufacturing method 4

[In the formula, _P1 , W, X, Y, Z, _R1 , _R2 , _R3 , and n have the same meanings as above.]

(Step 6) This step is to deprotect the amino group of the compound represented by general formula (IX) to produce the compound represented by general formula (X). The deprotection method can be a commonly known method, such as the method described in *Protective Groups in Organic Synthesis*, T.W. Greene, John Wiley & Sons (1981), or a method based thereon. A tert-butoxycarbonyl group can be used as a protecting group. When a tert-butoxycarbonyl group is used as the protecting group, deprotection under acidic conditions is preferred. Examples of acids include hydrochloric acid, acetic acid, trifluoroacetic acid, sulfuric acid, methanesulfonic acid, and toluenesulfonic acid. Alternatively, deprotection using a Lewis acid is also preferred; examples include trimethyliodosilane and boron trifluoride-diethyl ether coordination compounds. The amount of acid used is preferably 1 to 100 moles relative to 1 mole of compound (IX).

The solvent used in the reaction can be any solvent that does not adversely affect the reaction. Examples include alcohols (e.g., methanol), hydrocarbons (e.g., benzene, toluene, xylene), halogenated hydrocarbons (e.g., dichloromethane, chloroform, 1,2-dichloroethane), nitriles (e.g., acetonitrile), ethers (e.g., dimethoxyethane, tetrahydrofuran), aprotic polar solvents (e.g., N,N-dimethylformamide, dimethyl sulfoxide, hexamethylphosphoramide), or mixtures thereof. The reaction time is 0.1–100 hours, preferably 0.5–24 hours. The reaction temperature is 0–120°C, preferably 0–90°C.

The compound obtained in this way, represented by the general formula (X), can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc., or can be delivered to subsequent processes without separation and purification.

(Step 7) This step is a step of manufacturing the compound of the present invention represented by general formula (I) by reacting the compound represented by general formula (X) with a carboxylic acid represented by Y-COOH or an acyl halide represented by Y-C(=O)-L (L represents a chlorine atom or a bromine atom) through an amidation reaction.

When the carboxylic acid represented by Y-COOH is used as the amidation agent, in the presence of a suitable condensing agent, 0.5 to 10 moles, preferably 1 to 3 moles, of the carboxylic acid are used relative to 1 mole of the compound represented by general formula (X). This carboxylic acid can be commercially available or manufactured using known methods.

The reaction solvent can be any solvent that does not impede the reaction and is not particularly limited. Preferred solvents include isopropanol, tert-butanol, toluene, benzene, dichloromethane, chloroform, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, or mixtures thereof. The reaction temperature is typically -78 to 200°C, preferably 0 to 50°C. The reaction time is typically 5 minutes to 3 days, preferably 5 minutes to 10 hours.

Examples of condensing agents include diphenyl azidophosphate, N,N'-dicyclohexylcarbodiimide, benzotriazol-1-yloxy-tris-dimethylaminophosphonium salt, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, a combination of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 1-hydroxybenzotriazole, 2-chloro-1,3-dimethylimidazolium chloride, and O-(7-azobenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate.

Furthermore, a base may be added to the above reaction as needed. Examples of bases include organic bases such as triethylamine, diisopropylethylamine, pyridine, dimethylpyridine, trimethylpyridine, 4-dimethylaminopyridine, potassium tert-butyrate, sodium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, potassium hexamethyldisilazane, and butyllithium, or inorganic bases such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, and sodium hydride. The amount added is 1 to 100 moles, preferably 1 to 10 moles, relative to 1 mole of the compound represented by general formula (X).

When using an acyl halide represented by Y-C(=O)-L (where L represents a chlorine or bromine atom) as the amidation agent, 0.5 to 5 moles, preferably 0.9 to 1.1 moles, of the acyl halide are used relative to 1 mole of the compound represented by general formula (X). This acyl halide can be commercially available or manufactured using known methods.

The reaction solvent is not particularly limited as long as it does not hinder the reaction. Preferred solvents include water, toluene, benzene, dichloromethane, chloroform, tetrahydrofuran, acetonitrile, 1,4-dioxane, dimethylformamide, dimethylacetamide, N-methylpyrrolidone, or mixtures thereof. The reaction temperature is typically -78 to 200°C, preferably -20 to 50°C. The reaction time is typically 5 minutes to 3 days, preferably 5 minutes to 10 hours.

Furthermore, a base may be added to the above reaction as needed. Examples of bases include organic bases such as triethylamine, diisopropylethylamine, pyridine, dimethylpyridine, trimethylpyridine, 4-dimethylaminopyridine, potassium tert-butyrate, sodium tert-butyrate, sodium methoxide, sodium ethoxide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, potassium hexamethyldisilazane, and butyllithium, or inorganic bases such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, and sodium hydride. As the amount added, 1 to 100 moles, preferably 1 to 10 moles, may be used relative to 1 mole of the compound represented by general formula (X).

The compound represented by general formula (I) obtained in this way can be purified by known separation and purification methods, such as concentration, vacuum concentration, crystallization, solvent extraction, reprecipitation, chromatography, etc.

Manufacturing Method 5

(Process 8, Process 9)

This process is the same as manufacturing method 4, process 6, and process 7, which involves manufacturing the compound of general formula (XII) from the compound of general formula (VIII).

(Process 10)

This process is the same as manufacturing method 2 and process 4, which involves manufacturing the compound of general formula (I) from the compound of general formula (XII).

In the manufacturing methods 1 to 5 described above, functional groups such as amino, imino, hydroxyl, carboxyl, carbonyl and amide groups, as well as indole, which have active protons, can be protected by a reagent in appropriate steps of each manufacturing method, or the functional group can be introduced with a protecting group according to conventional methods and then the protecting group can be removed.

As a "protecting group for amino or imino groups," any group that performs this function is acceptable and is not particularly limited. Examples include aralkyl groups such as benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, diphenylmethyl, triphenylmethyl, and cumyl; lower alkyl acyl groups such as formyl, acetyl, propionyl, butyryl, p-valeryl, trifluoroacetyl, and trichloroacetyl; benzoyl; arylalkyl acyl groups such as phenylacetyl and phenoxyacetyl; lower alkyloxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, and tert-butoxycarbonyl; and p-nitrobenzyloxycarbonyl. Aryl alkoxy carbonyl groups such as phenylethoxycarbonyl; lower alkyl silyl groups such as trimethylsilyl, tert-butyldimethylsilyl; tetrahydropyranyl; trimethylsilylethoxymethyl; lower alkyl sulfonyl groups such as methylsulfonyl, ethylsulfonyl, tert-butylsulfonyl; lower alkyl sulfonyl groups such as tert-butylsulfinyl; aryl sulfonyl groups such as benzenesulfonyl, toluenesulfonyl; imide groups such as phthalimide, especially preferably trifluoroacetyl, acetyl, tert-butyloxycarbonyl, benzyloxycarbonyl, trimethylsilylethoxymethyl, cumyl, etc.

As a "protecting group for the hydroxyl group," any group that has this function is acceptable and there are no particular limitations. Examples include lower alkyl groups such as methyl, ethyl, propyl, isopropyl, and tert-butyl; lower alkylsilyl groups such as trimethylsilyl and tert-butyldimethylsilyl; lower alkoxymethyl groups such as methoxymethyl and 2-methoxyethoxymethyl; tetrahydropyranyl; trimethylsilylethoxymethyl; aralkyl groups such as benzyl, p-methoxybenzyl, 2,3-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, and triphenylmethyl; and acyl groups such as formyl, acetyl, and trifluoroacetyl. Methyl, methoxymethyl, tetrahydropyranyl, trimethylsilylethoxymethyl, tert-butyldimethylsilyl, and acetyl are particularly preferred.

As a "protecting group for the carboxyl group," any group that has this function is acceptable and there are no particular limitations. Examples include lower alkyl groups such as methyl, ethyl, propyl, isopropyl, and tert-butyl; lower alkyl halogens such as 2,2,2-trichloroethyl; lower alkenyl groups such as allyl; trimethylsilylethoxymethyl; and aralkyl groups such as benzyl, p-methoxybenzyl, p-nitrobenzyl, diphenylmethyl, and triphenylmethyl. Methyl, ethyl, tert-butyl, allyl, benzyl, p-methoxybenzyl, and trimethylsilylethoxymethyl are particularly preferred.

As a "protecting group of the carbonyl group", it is acceptable as long as it has this function, without any particular limitation. For example, ethylene glycol ketal, propylene glycol ketal, dimethyl ketal, acetal, propional acetal, dimethyl acetal, and other ketals and acetals can be listed.

As a "protecting group of a functional group with an active proton, such as an amide group or an indole group", it is acceptable as long as it has this function, without particular limitation. Examples include lower alkyl groups such as methyl, ethyl, propyl, isopropyl, and tert-butyl; lower alkylsilyl groups such as trimethylsilyl and tert-butyldimethylsilyl; lower alkoxymethyl groups such as methoxymethyl and 2-methoxyethoxymethyl; tetrahydropyranyl; trimethylsilylethoxymethyl; aralkyl groups such as benzyl, p-methoxybenzyl, 2,3-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, and triphenylmethyl; and acyl groups such as formyl, acetyl, and trifluoroacetyl. Methyl, methoxymethyl, tetrahydropyranyl, trimethylsilylethoxymethyl, tert-butyldimethylsilyl, and acetyl are particularly preferred.

The method for removing the protecting group varies depending on the type of protecting group and the stability of the target compound. For example, it can be carried out according to the methods described in the literature (see Protective Groups in Organic Synthesis, 3rd edition, by T.W. Greene, John Wiley & Sons, 1999) or equivalent methods. For example, it can be carried out by solvent decomposition using acid or base, i.e., by using 0.01 moles to a large excess of acid, preferably trifluoroacetic acid, formic acid, hydrochloric acid, etc., or equimolar to a large excess of base, preferably potassium hydroxide, calcium hydroxide, etc.; chemical reduction using hydride metal coordination compounds, etc., or contact reduction using palladium-carbon catalysts, Lanney nickel catalysts, etc.

The compounds of the present invention can be readily purified by conventional separation methods. Examples of such methods include solvent extraction, recrystallization, reversed-phase high-speed liquid chromatography for separation, column chromatography, and thin-layer chromatography for separation.

When the compounds of the present invention have isomers such as optical isomers, stereoisomers, positional isomers, and rotational isomers, mixtures of any of the isomers are also included in the compounds of the present invention. For example, when the compounds of the present invention contain optical isomers, the optical isomers obtained by resolution of the racemic mixture are also included in the compounds of the present invention. These isomers can be obtained in the form of single compounds by known synthetic methods and separation methods (concentration, solvent extraction, column chromatography, recrystallization, etc.).

The compounds of the present invention or their salts may be crystalline, whether in a single crystal form or a mixture of polymorphs. Crystals can be produced by crystallization using methods known per se. The compounds of the present invention or their salts may be solvates (e.g., hydrates) or solvent-free compounds, both of which are included in the compounds of the present invention or their salts. Compounds labeled with isotopes (e.g., ^3H , ^14C , ^35S , ^125I , etc.) are also included in the compounds of the present invention or their salts.

The precursor of the compound or its salt of the present invention refers to a compound that is converted into the compound or its salt of the present invention under physiological conditions in vivo through a reaction utilizing enzymes or gastric acid, etc. Specifically, it refers to a compound that is converted into the compound or its salt of the present invention through enzyme-catalyzed oxidation, reduction, hydrolysis, etc., or a compound that is converted into the compound or its salt of the present invention through hydrolysis utilizing gastric acid, etc. Furthermore, the precursor of the compound or its salt of the present invention can be converted into the compound or its salt of the present invention under the physiological conditions described in the 1990 publication *Medical Drug Development*, Volume 7, Molecular Design, pages 163-198, by Hirokawa Shoten.

The salts of the compounds of this invention refer to salts commonly used in the field of organic chemistry, such as salts that are base addition salts of the carboxyl group when it has a carboxyl group, or salts that are acid addition salts of the amino or basic heterocyclic group when it has an amino or basic heterocyclic group.

Examples of alkali metal salts that can be used as base addition salts include sodium and potassium salts; alkaline earth metal salts such as calcium and magnesium salts; ammonium salts; and organic amine salts such as trimethylamine salts, triethylamine salts, dicyclohexylamine salts, ethanolamine salts, diethanolamine salts, triethanolamine salts, procaine salts, and N,N'-dibenzylethylenediamine salts.

Examples of acid addition salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate, and perchlorate; organic acid salts such as acetate, formate, maleate, fumarate, tartrate, citrate, ascorbate, and trifluoroacetate; and sulfonates such as methanesulfonate, hydroxyethylsulfonate, benzenesulfonate, and p-toluenesulfonate.

The compounds of the present invention, or their salts, possess excellent BTK inhibitory activity and are useful as preventive and/or therapeutic agents for various immune diseases, such as allergic diseases, autoimmune diseases, and inflammatory diseases, particularly allergic or autoimmune diseases. Furthermore, they exhibit excellent selectivity for BTK, with the advantage of fewer side effects caused by the inhibition of other kinases (e.g., EGFR).

In this specification, "BTK" includes BTK from humans or non-human mammals, preferably human BTK. Furthermore, the term "BTK" includes subtypes.

In addition, the compounds of the present invention or their salts, due to their excellent BTK inhibitory activity, are useful as medicines for the prevention or treatment of BTK-involved immune diseases, such as allergic diseases, autoimmune diseases, inflammatory diseases, especially allergic diseases or autoimmune diseases.

"BTK-involved immune diseases" can be listed as diseases that reduce morbidity, alleviate, mitigate, and/or cure symptoms by causing loss of BTK function, suppression, and/or obstruction.

The immune diseases that are the subject of the study can be any immune diseases caused by abnormal expression and/or activity of BTK, without any particular restrictions. Examples include allergic diseases, autoimmune diseases, and inflammatory diseases.

Existing rheumatoid arthritis drugs (such as the pan-JAK inhibitor Tofacitnib) have been shown to reduce the host's immune capacity by decreasing the number of NK cells, and pose a high risk of herpes virus infection or cancer (ACRHOTILINE, December 14, 2012).

The compounds or their salts of the present invention have high cell selectivity and low inhibitory effect on NK cells, thus possessing the advantages of low risk of host immunosuppression and high safety.

Furthermore, the compounds of the present invention or their salts can inhibit bone resorption by osteoclasts in mice and restore bone density, thus they are also useful for osteoclast-related diseases (such as osteoporosis).

There are no particular restrictions on the allergic diseases that can be included, such as bronchial asthma, allergic rhinitis, pollen allergy, atopic dermatitis, food allergy, anaphylaxis, drug allergy, urticaria, and conjunctivitis. Bronchial asthma, allergic rhinitis, pollen allergy, and atopic dermatitis are preferred, with allergic rhinitis, pollen allergy, and atopic dermatitis being particularly preferred.

There are no particular restrictions on the autoimmune diseases that can be included; examples include rheumatoid arthritis, systemic lupus erythematosus, scleroderma, polymyositis, Sjögren's syndrome, and Behcet's disease. Rheumatoid arthritis and systemic lupus erythematosus are preferred, with rheumatoid arthritis being particularly preferred.

There are no particular limitations on the inflammatory diseases that can be included, such as appendicitis, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, ulcerative colitis, Crohn's disease, irritable bowel disease, cystitis, dacryoadenitis, contact dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, epididymitis, fasciitis, fibrositis, gastroenteritis, hepatitis, hidradenitis, laryngitis, mastitis, meningitis, spinal cord inflammation, myocarditis, nephritis, oophoritis, ovarian inflammation, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, pneumonia, proctitis, prostatitis, pyelonephritis, otitis media, sinusitis, stomatitis, osteoarthritis, synovitis, tendinitis, tonsillitis, uveitis, vaginitis, vasculitis, and vulvitis. Ulcerative colitis, Crohn's disease, irritable bowel disease, contact dermatitis, cystitis, and osteoarthritis are preferred. Contact dermatitis, cystitis, and osteoarthritis are especially preferred.

The preferred immune diseases to be treated are allergic diseases and autoimmune diseases, with atopic dermatitis, rheumatoid arthritis, systemic lupus erythematosus, allergic rhinitis or hay fever being more preferred, and rheumatoid arthritis being particularly preferred.

When formulating the compounds of the present invention or their salts into pharmaceuticals, a drug carrier may be used as needed. Various administration forms may be employed depending on the purpose of prevention or treatment, such as oral dosage forms, injections, suppositories, ointments, patches, etc. These administration forms can be manufactured using various formulation methods known and commonly used by those skilled in the art.

As a drug carrier, various organic or inorganic carrier substances commonly used as raw materials in pharmaceutical preparations can be used. In solid formulations, they can be combined with excipients, binders, disintegrants, lubricants, coating agents, etc., while in liquid formulations, they can be combined with solvents, solubilizers, suspending agents, isotonic agents, pH adjusters/buffers, analgesics, etc. Furthermore, preservatives, antioxidants, colorants, flavoring/odor-correcting agents, stabilizers, and other pharmaceutical additives can be used as needed.

Examples of excipients include lactose, white sugar, D-mannitol, starch, crystalline cellulose, and calcium silicate.

Examples of binders include hydroxypropyl cellulose, methylcellulose, polyvinylpyrrolidone, powdered sugar, and hydroxypropyl methylcellulose.

Examples of disintegrants include sodium glycolate starch, calcium carboxymethyl cellulose, croscarmellose sodium, cross-linked polyvinyl cellulose, low-substituted hydroxypropyl cellulose, and partially α-substituted starch.

Examples of lubricants include talc, magnesium stearate, sucrose fatty acid esters, stearic acid, and sodium stearate fumarate.

Examples of coating agents include ethyl cellulose, aminoalkyl methacrylate copolymer RS, hydroxypropyl methylcellulose, and white sugar.

Examples of solvents include water, propylene glycol, and physiological saline.

Examples of co-solvents include polyethylene glycol, ethanol, α-cyclodextrin, polyethylene glycol 400 (Macrogol 400), and polysorbate 80.

Examples of suspending agents include carrageenan, crystalline cellulose-sodium carboxymethyl cellulose, and polyoxyethylene hydrogenated castor oil.

Examples of isotonic agents include sodium chloride, glycerol, and potassium chloride.

Examples of pH adjusters/buffers include sodium citrate, hydrochloric acid, lactic acid, phosphoric acid, and sodium dihydrogen phosphate.

Examples of analgesics include procaine hydrochloride and lidocaine.

Examples of preservatives include ethylparaben, cresol, and benzalkonium chloride.

Examples of antioxidants include sodium sulfite, ascorbic acid, and natural vitamin E.

Examples of colorants include titanium dioxide, ferric oxide, Edible Blue No. 1, and copper chlorophyll.

Examples of flavoring and odor-correcting agents include aspartame, saccharin, sucralose, L-menthol, and peppermint flavoring.

Examples of stabilizers include sodium metabisulfite, sodium edetate, isoascorbic acid, magnesium oxide, and butylated hydroxytoluene.

When preparing oral solid dosage forms, excipients, binders, disintegrants, lubricants, colorants, flavoring/odor-correcting agents, etc., can be added to the compounds of this invention as needed, and then tablets, coated tablets, granules, powders, capsules, etc. can be manufactured according to conventional methods.

In preparing injectable formulations, pH adjusters, buffers, stabilizers, isotonic agents, local anesthetics, etc., can be added to the compounds of this invention to manufacture subcutaneous, intramuscular, and intravenous injectable formulations according to conventional methods.

The amount of the compound of the present invention to be used in each of the above-mentioned dosing unit forms cannot be generalized depending on the symptoms of the patient to whom the compound is to be used or the dosage form. Generally speaking, the expected dosage per dosing unit form is 0.05 to 1000 mg for oral administration, 0.01 to 500 mg for injection, and 1 to 1000 mg for suppositories.

In addition, the daily dosage of the drug having the above-mentioned administration form varies depending on the patient's symptoms, weight, age, gender, etc., and cannot be generalized. Based on the compound of the present invention, the usual adult (weight 50kg) dose is 0.05 to 5000 mg per day, preferably 0.1 to 1000 mg, preferably administered once a day or divided into 2 to 3 doses.

Example

The present invention will now be described in more detail by way of examples, but the present invention is not limited to these examples at all.

Unless otherwise specified, all reagents used in the examples are commercially available. Silica column chromatography used Moritex Corp.'s PURIF-PACK SI, Biotage's KP-Sil Silica pre-packed column, or Biotage's HP-Sil Silica pre-packed column. Basic silica column chromatography used Moritex Corp.'s PURIF-PACK NH or Biotage's KP-NH pre-packed column. Thin-layer chromatography for separation used Merck's Kieselgel TM 60F254, Art. 5744, or WakoPure Chemical Industries, Ltd.'s NH2 silica gel 60F254 plates. NMR spectra were obtained using an AL400 (400MHz, JEOL), Mercury400 (400MHz, Agilent Technologies, Inc.) spectrometer, or an Inova400 (400MHz, Agilent Technologies, Inc.) spectrometer equipped with an OMNMR probe (Protasis). When the deuterated solvent contained tetramethylsilane, tetramethylsilane was used as an internal standard; otherwise, the NMR solvent was used as an internal standard. All δ values are expressed in ppm. Microwave reactions were performed using a Discover S-class microwave reactor manufactured by CEM.

In addition, LCMS spectra were determined using a Waters ACQUITY SQD (quadripolar) spectrometer under the following conditions.

Column: YMC-Triart C18, 2.0 x 50 mm, 1.9 μm, manufactured by YMC Corporation.

MS test: ESI positive

UV detection: 254 and 210nm

Column flow rate: 0.5 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 1 μL

Gradient (Table 1)

[Table 1]

Time (minutes)	water	Acetonitrile
			0	95	5
0.1	95	5
			2.1	5	95
3.0	Stop

In addition, reversed-phase separation HPLC purification was performed using a separation system manufactured by WATERS under the following conditions.

Column: A column made by connecting YMC-Actus Triart C18, 20×50mm, 5μm and YMC-Actus Triart C18, 20×10mm, 5μm manufactured by YMC.

UV detection: 254nm

MS test: ESI positive

Column flow rate: 25 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 0.1–0.5 mL

The meanings of the abbreviation symbols are as follows.

s: single peak

d: double peak

t: triple peak

q: Four Peaks

dd: Double double peak

dt: double and triple peaks

td: triple and double peaks

tt: Double Triple Peak

ddd: double double double peak

ddt: doublet doublet triplet peak

dtd: double triple double peak

tdd: triple double double peak

m: multiple peaks

br: broad peak

brs: broad single peak

CDI: Carbonyldiimidazole

DMSO- _d6 : Deuterated dimethyl sulfoxide

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

THF: Tetrahydrofuran

DMF: N,N-Dimethylformamide

DMA: N,N-dimethylacetamide

NMP: 1-Methyl-2-pyrrolidone

DMSO: Dimethyl sulfoxide

TFA: Trifluoroacetic acid

WSC: 1-(3-dimethylaminopropyl)-3- and carbodiimide hydrochloride

HOBt: 1-Hydroxybenzotriazole hydrate

HATU: (dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazol[4,5-b]pyridin-3-yloxy)methaneimine hexafluorophosphate

DIAD: Diisopropyl azodicarbonate

TBAF: Tetrabutylammonium fluoride

DIPEA: Diisopropylethylamine

Boc: tert-Butoxycarbonyl

Boc ₂ O: Ditert-butyl dicarbonate

DMAP: Dimethylaminopyridine

Synthetic Example 1 (R) - tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl) Synthesis of piperidine-1-carboxylic acid esters

(Step 1) Synthesis of (S)-tert-butyl-3-(methylsulfonyloxy)piperidine-1-carboxylic acid ester

20 g of (S)-N-Boc-3-piperidinol was dissolved in 100 mL of toluene. 21 mL of triethylamine and 9.2 mL of methanesulfonyl chloride were added at 0 °C. After stirring for 1 hour under ice-cold conditions, ethyl acetate and water were added, and the organic layer was separated. The organic layer was washed with saturated aqueous sodium bicarbonate solution, saturated ammonium chloride solution, and water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation to obtain 26.8 g of the title compound as a colorless solid.

(Step 2) Synthesis of (R)-tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

A suspension of 14.6 g of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine, synthesized according to the method described in International Publication No. 2007/126841, 25 g of (S)-tert-butyl-3-(methanesulfonyloxy)piperidine-1-carboxylic acid ester obtained in step 1, and 69 g of potassium carbonate in 150 mL of DMA was heated to 100 °C and stirred for 10 hours. After cooling to room temperature, 300 mL of water was added, the resulting solid was filtered to separate the solid, washed with water, and dried to obtain 26.9 g of the title compound as a yellow solid.

Physical property value: m/z[M+H] ⁺ 446.2

Synthetic Example 2 (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3, Synthesis of 4-d]pyrimidine-3-carboxylic acid

2 g of (R)-tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester, 3 mL of ₂ -diethylaminoethanol, and 158 mg of Pd( _PPh3 ) _2Cl2 obtained in Synthetic Example 1 were dissolved in 20 mL of NMP. After purging the system with carbon monoxide, the mixture was heated to 120 °C. After stirring for 1 hour, the mixture was cooled to room temperature, and 10 mL of methanol was added. Then, 6 mL of 5N sodium hydroxide aqueous solution was added, and the mixture was stirred for 10 minutes. After adding water, the aqueous layer was washed with ethyl acetate, and the pH of the aqueous layer was adjusted to 4 with hydrochloric acid. The precipitated solid was separated by filtration, washed with water, and dried to obtain 1.26 g of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 363.1

Synthetic Example 3: (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolidine Synthesis of [3,4-d]pyrimidine-3-carboxylic acid

(Step 1) Synthesis of (S)-tert-butyl-3-(methylsulfonyloxy)pyrrolidine-1-carboxylic acid ester

935 mg of (S)－(－)－N－Boc－3－pyrrolidone was dissolved in 15 mL of chloroform. Under ice-cold conditions, 1.04 mL of triethylamine and 467 μL of methanesulfonyl chloride were added. After stirring at room temperature for 1.5 hours, ethyl acetate and water were added, and the organic layer was separated. The organic layer was washed with saturated aqueous sodium bicarbonate solution, saturated aqueous sodium chloride solution, and water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation to obtain 1.3 g of the title compound as a colorless oil.

Physical property value: m/z[M+H] ⁺ 266.1

(Step 2) Synthesis of (R)-tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester

20.0 g of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine, synthesized according to the method described in International Publication No. 2007/126841, 23 g of (S)-tert-butyl-3-(methanesulfonyloxy)pyrrolidine-1-carboxylic acid ester obtained in step 1, and 32 g of potassium carbonate were suspended in 200 mL of DMA. The suspension was heated to 85 °C and stirred for 3 hours. After cooling to room temperature, 400 mL of water was added, and the resulting solid was filtered, washed with water, and dried to obtain 23.5 g of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 431.0

(Step 3) Synthesis of (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid

2.0 g of (R)-tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester, 3.1 mL of ₂ -diethylaminoethanol, and 163 mg of Pd( _PPh3 ) _2Cl2 obtained in step 2 above were dissolved in 20 mL of NMP. After purging the system with carbon monoxide, the mixture was heated to 120 °C. After stirring for 1 hour, the mixture was cooled to room temperature, 10 mL of methanol was added, followed by 6 mL of 5N sodium hydroxide aqueous solution, and the mixture was stirred for 10 minutes. After adding water, the aqueous layer was washed with chloroform, and the pH of the aqueous layer was adjusted to 4 with hydrochloric acid. The precipitated solid was filtered and separated, washed with water, and dried to obtain 1.35 g of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 349.1

Synthesis Example 4: Synthesis of 5-cyanobenzo[d]oxazol-2-amine

15.1 g of 3-amino-4-hydroxybenzonitrile was dissolved in 75 mL of ethanol and 75 mL of water. 14.7 g of cyanogen bromide was added dropwise to the solution while ice-cold. The mixture was stirred at room temperature for 2 hours and then cooled again. 112 mL of 2N sodium hydroxide aqueous solution was added to the solution, and the mixture was stirred for another 30 minutes. The ethanol was largely removed using an evaporator, and the residue was separated by filtration. The filtered residue was washed with water to give 12.12 g of the title compound.

Physical property value: m/z[M+H] ⁺ 161.1

Example 1 (R)-1-(1-acryloylpiperidin-3-yl)-4-amino-N-(5-chlorobenzo[d]oxa) Synthesis of (-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 1)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

In Synthesis Example 2, 94 mg of (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid was added to a suspension of 94 mg in 4 mL of THF, and CDI was added. The mixture was stirred at room temperature for 3 hours. 66 mg of 5-chlorobenzo[d]oxazol-2-amine was added under ice-cold conditions, followed by dropwise addition of a 1.0 M solution of lithium hexamethyldisilazane-THF. After stirring under ice-cold conditions for 30 minutes, 1 mL of water was added to remove the THF solvent. 4 mL of water was added to the residue, and the resulting solid was filtered and washed with 1/1.5 mL of hexane/ethyl acetate to give 106 mg of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 513.2

(Step 2) Synthesis of Compound 1 in Example

5.6 mg of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in (step 1) was added to 1 mL of 4N hydrochloric acid/1,4-dioxane. After stirring for 1 hour, the solvent was removed by evaporation. 2 mL of chloroform and 7.6 μL of triethylamine were added to the residue, and after cooling, 0.9 μL of acryloyl chloride was added. After stirring for 1.5 h, the reaction was stopped using a saturated sodium bicarbonate aqueous solution, and the mixture was extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by silica gel column chromatography (development solvent: ethyl acetate/methanol) to give 2.6 mg of the title compound as a white solid.

Example 2 (R)-1-(1-acryloylpiperidin-3-yl)-4-amino-N-(5-bromobenzo[d]oxa Synthesis of (-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 2)

According to Example 1, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-bromobenzo[d]oxazol-2-amine of Synthetic Example 2.

Example 3 (R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(5-(thiophene-2-) The synthesis of (-)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 3) become

(Step 1) Synthesis of 5-(thiophene-2-yl)benzo[d]oxazol-2-amine

100 mg of 5-bromobenzo[d]oxazol-2-amine, 249 mg of potassium phosphate, and 90 mg of thiophene-2-ylboronic acid were suspended in 2.5 mL of DME and 0.5 mL of water. 38 mg of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)-dichloromethane was added, and the mixture was heated at 140 °C for 20 minutes in a microwave reactor. The solvent was removed from the reaction solution, and the solution was purified by amine gel chromatography (dissolution buffer: chloroform/methanol) to give 93 mg of the title compound as a light brown solid.

Physical property value: m/z[M+H] ⁺ 216.8

(Step 2) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-(thiophen-2-yl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

In Synthesis Example 2, 19 mg of (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid was added to a suspension of 19 mg in 2 mL of THF, and the mixture was stirred at room temperature for 2 hours. Under ice-cold conditions, 17 mg of 5-(thiophene-2-yl)benzo[d]oxazol-2-amine obtained in Step 1 was added, followed by dropwise addition of 105 μL of hexamethyldisilazane lithium salt-THF 1.0 M solution. After stirring under ice-cold conditions for 30 minutes, 1 mL of water was added to remove the THF solvent. 4 mL of water was added to the residue, and the resulting solid was filtered and washed with 5 mL of a 1/1 mixture of hexane/ethyl acetate to obtain 13 mg of the target substance as a light brown solid.

Physical property value: m/z[M+H] ⁺ 561.3

(Step 3) Synthesis of Compound 3 in Example

In step 2, 9 mg of (R)-tert-butyl-3-(4-amino-3-((5-(thiophen-2-yl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane. After stirring for 1 hour, the solvent was removed by evaporation. 2 mL of chloroform and 12 μL of triethylamine were added to the residue, and after cooling, 1.3 μL of acryloyl chloride was added. After stirring for 1.5 h, the reaction was stopped with saturated sodium bicarbonate aqueous solution, and extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 2.1 mg of the title compound as a white solid.

Example 4 (R)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-methacryloyl) Synthesis of pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 4)

According to Example 1, methacryloyl chloride was used instead of acryloyl chloride to obtain the title compound as a white solid.

Example 5 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-yl) Synthesis of (enyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 5)

According to Example 1, butenoyl chloride was used instead of acryloyl chloride to obtain the title compound as a white solid.

Example 6 (R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(5-cyanobenzo[d]) Synthesis of Oxazo-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 6)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-cyanobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

2.32 g of (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid obtained in Synthetic Example 2 was dissolved in 25 mL of DMA, and 2.01 g of CDI was added. The mixture was stirred at room temperature for 1 hour. 1.12 g of 5-cyanobenzo[d]oxazol-2-amine was added to the reaction solution, followed by 1.23 g of sodium tert-butyrate. The mixture was stirred at room temperature for 2 hours, and after adding water, the pH was adjusted with 2N hydrochloric acid to precipitate the solid. The solid was filtered, separated, and dried to give 2.66 g of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 505.3

(Step 2) Synthesis of Compound 6 in Example

2.1 g of (R)-tert-butyl-3-(4-amino-3-((5-cyanobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in step 1 was suspended in 10 mL of dichloromethane, and 10 mL of TFA was added at room temperature. After stirring for 2 hours, the solvent TFA was removed by evaporation. Further azeotropic reaction was carried out using toluene, and 20 mL of NMP and 2 mL of water were added to the residue, followed by ice cooling. 2.88 g of potassium carbonate and 0.4 mL of acryloyl chloride were added, and the mixture was stirred while ice-cold. After 2 hours, water and 2N hydrochloric acid were added to adjust the pH, and the resulting solid was separated by filtration. Purification was then performed using silica gel chromatography (chloroform-methanol) to obtain 0.7 g of the target substance as a white solid.

Example 7 (R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(5-methoxybenzoxyl) Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 7)

According to Example 6, the title compound, a light brown solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-methoxybenzo[d]oxazol-2-amine of Synthetic Example 2.

Example 8 (R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(5-(2-methoxyethyl) The synthesis of (-)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 8) become

According to Example 6, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-(2-methoxyethyl)benzo[d]oxazol-2-amine of Synthetic Example 2.

Example 9 (R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(oxazolo[4,5-b]) Synthesis of pyridin-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 9)

According to Example 6, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and oxazolo[4,5-b]pyridine-2-amine of Synthetic Example 2.

Example 10(R)-1-(1-Acryloylpiperidin-3-yl)-4-amino-N-(4-methylbenzo[d]) Synthesis of Oxazo-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 10)

According to Example 6, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 4-methylbenzo[d]oxazol-2-amine of Synthetic Example 2.

Example 11(R)-4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)-1-(1-methacryloyl) Synthesis of pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 11)

According to Example 6, using (R)-4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide obtained in Example 12 (step 2) and methacryloyl chloride instead of acryloyl chloride, the title compound, a white solid, was obtained.

Example 12(R)-1-(1-acryloylpiperidin-3-yl)-4-amino-N-(5-fluorobenzo[d]oxa) Synthesis of (-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 12)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-fluorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

In a solution of (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid 1.0 g in 10 mL of DMA, 895 mg of CDI was added, and the mixture was stirred at room temperature for 1 hour. 462 mg of 5-fluorobenzo[d]oxazol-2-amine was added, followed by 9 mL of sodium tert-butyrate-THF 1.0 M solution. After stirring at room temperature for 30 minutes, 10 mL of 1N sodium hydroxide aqueous solution was added to remove the THF solvent. After stirring for 1 hour, crystals were precipitated with 2N hydrochloric acid, and water/methanol was added to ensure complete crystallization. The resulting solid was then filtered to obtain 1.14 g of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 497.2

(Step 2) Synthesis of (R)-4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide

3.06 g of (R)-tert-butyl-3-(4-amino-3-(5-fluorobenzo[d]oxazol-2-ylcarbonyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in step 1 and 5.5 g of sodium iodide were suspended in 30 mL of acetonitrile, and 4.7 mL of trimethylchlorosilane was added at room temperature. The mixture was stirred at room temperature for 1 hour, and then a saturated aqueous solution of sodium bicarbonate was added to precipitate the solid. After stirring for 10 minutes, the solid was filtered and dried to obtain 2.07 g of the title compound as a light brown solid.

Physical property value: m/z[M+H] ⁺ 398.0

(Step 3) Synthesis of Compound 12 in Example

2 g of (R)-4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide obtained in step 2 and 2.1 g of potassium carbonate were dissolved in 20 mL of NMP and 2 mL of water, and stirred under ice-cold conditions. 0.4 mL of acryloyl chloride was added and stirred for 1 hour. Water was added, and the pH was adjusted with hydrochloric acid. The precipitated solid was separated by filtration. The filtered solid was purified by silica gel chromatography (dissolution buffer: chloroform/methanol) to obtain 1.79 g of the title compound as a white solid.

Example 13(R)-1-(1-acryloylpiperidin-3-yl)-4-amino-N-(benzo[d]oxazol- Synthesis of 2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 13)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

300 mg of (R)-tert-butyl-3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in Synthetic Example 1 was dissolved in 3 mL of NMP. 118 mg of benzo[d]oxazol-2-amine, 20 mg of 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene, and 0.15 mL of N-methylmorpholine were added, followed by degassing. Then, 7.6 mg of palladium acetate was added, and the mixture was heated to 110 °C and stirred for 2 hours in a carbon monoxide atmosphere. After cooling, 4.5 mL of methanol and 0.45 mL of 5N sodium hydroxide aqueous solution were added, and the mixture was stirred at room temperature for 30 minutes. The pH was then adjusted to 5.3 with 2N hydrochloric acid, and the resulting solid was separated by filtration. The crude product was purified by silica gel column chromatography (chloroform/methanol) to give 257 mg of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 479.3

(Step 2) Synthesis of Compound 13 in Example

5 g of (R)-tert-butyl-3-(4-amino-3-((benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in step 1 was suspended in 50 mL of acetonitrile, and 7.85 g of sodium iodide was added. 6.65 mL of trimethylchlorosilane was added dropwise while stirring at room temperature, and the mixture was stirred for 1 hour. 87.5 mL of water and 12.5 mL of 5N sodium hydroxide aqueous solution were added, and the mixture was chilled. 0.895 mL of acryloyl chloride was added dropwise, and the mixture was stirred for 1 hour while chilled. Water was added, and the resulting solid was filtered, washed with water, and dried to obtain 4.13 g of the title compound as a white solid.

Example 14 (R, E)-4-amino-N-(benzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl) Synthesis of (3,4-d)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 14)

According to Example 13, butenoyl chloride was used instead of acryloyl chloride to obtain the title compound as a white solid.

Example 15 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(di) (Methylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example) Synthesis of compound 15

In step 1 of Example 1, 5 mg of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane and stirred for 10 minutes. The solvent was then removed by evaporation and azeotropically treated with toluene. The residue was dissolved in 1 mL of DMF, and 8.5 μL of DIPEA, 2.4 mg of (E)-4-(dimethylamino)but-2-enoate, and 5.5 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 3.96 mg of the title compound as a white solid.

Example 16 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(ethyl) (methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (actual) Synthesis of Example Compound 16

According to Example 15, (E)-4-(ethyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 17 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(di) (Ethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example) Synthesis of compound 17

According to Example 15, (E)-4-(diethylamino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 18 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(iso) Propyl(methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide Synthesis of (Example Compound 18)

According to Example 15, (E)-4-(isopropyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 19 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyridine) (Pyrazolyl-1-yl)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (actual) Synthesis of Example Compound 19)

According to Example 15, (E)-4-(pyrrolidine-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 20 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(piperazolyl) ... (Pyridin-1-yl)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (implementation) Synthesis of compound 20 (Example)

According to Example 15, (E)-4-(piperidin-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 21 (R, E)-4-amino-N-(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)-1- (1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-methyl Synthesis of amide (Compound 21 in Example)

According to Example 15, using (R)-tert-butyl-3-(4-amino-3-((5-(thiophen-2-yl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in Example 3 (step 2), the title compound, a white solid, was obtained.

Example 22(R)-4-amino-N-(benzo[d]oxazol-2-yl)-1-(1-(but-2-ynyl)) Synthesis of piperidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 22)

According to Example 15, using (R)-tert-butyl-3-(4-amino-3-((benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in Example 13 (step 1) and butyrynic acid substituted for (E)-4-(dimethylamino)butyrylic acid ester, the title compound, a pale yellow solid, was obtained.

Example 23(R)-1-(1-acryloylpiperidin-3-yl)-4-amino-N-(5,6-dimethylbenzene) Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 23)

(Step 1) Synthesis of (R)-1-(1-acyloxypiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid

In Synthesis Example 2, 1 g of (R)-4-amino-1-(1-(tert-butoxycarbonyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid was added to 15 mL of 4N hydrochloric acid/1,4-dioxane, and the mixture was stirred at room temperature for 1 hour. The solvent was then removed, and toluene was added for azeotropic reaction. 50 mL of chloroform and 3.8 mL of triethylamine were added to the residue. Acryloyl chloride (780 μL) was slowly added while stirring. After confirming the reaction was complete, 2-propanol was added to stop the reaction. The solvent was removed, and an aqueous formic acid solution was added to the residue to adjust the pH to 3, at which point a solid precipitated. The solid was filtered, dried, and yielded 840 mg of the title compound as a yellow solid.

Physical property value: m/z[M+H] ⁺ 318.1

(Step 2) Synthesis of Compound 23 in Example

Dissolve 5 mg of (R)-1-(1-acyloxypiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid obtained in step 1 above in 150 μL of DMF. In this solution, add 8.26 μL of diisopropylethylamine, 3.85 mg of 5,6-dimethylbenzo[d]oxazol-2-amine, and 9 mg of HATU. After stirring overnight, add 850 μL of DMSO and purify by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to obtain 1.2 mg of the title compound as a white solid.

Example 24(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-chlorobenzo[d]) Synthesis of Oxazo-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 24)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyridine-1-carboxylic acid ester

In a solution of (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid 100 mg in DMF 5 mL, 56 mg of CDI was added and the mixture was stirred at room temperature for 1 hour.

73 mg of 5-chlorobenzo[d]oxazol-2-amine and 17 mg of 60% sodium hydride were added under ice-cold conditions. After stirring for 30 minutes under ice-cold conditions, 1 mL of water was added to stop the reaction. The reaction solution was concentrated and purified by silica gel column chromatography (developing solvent: chloroform/methanol) to give 114 mg of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 499.1

(Step 2) Synthesis of Compound 24 in Example

15 mg of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester obtained in step 1 was added to 1.5 mL of 4N hydrochloric acid/1,4-dioxane. After stirring for 1 hour, the solvent was removed by evaporation. 2 mL of chloroform and 21 μL of triethylamine were added to the residue, and after cooling, 2.4 μL of acryloyl chloride was added. After stirring for 3 hours, the reaction was stopped with saturated sodium bicarbonate aqueous solution, and extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by silica gel column chromatography (developing solvent: ethyl acetate/methanol) to give 6.8 mg of the title compound as a white solid.

Example 25 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-yl) The synthesis of (enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 25) become

According to Example 24, butenoyl chloride was used instead of acryloyl chloride to obtain the title compound as a white solid.

Example 26 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(3-methyl) (but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound) Synthesis of 26)

According to Example 24, 3-methylbut-2-enoyl chloride was used instead of acryloyl chloride to obtain the title compound as a white solid.

Example 27 (R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(benzo[d]oxa) Synthesis of (-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 27)

According to Example 24, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and benzo[d]oxazol-2-amine of Synthetic Example 3.

Example 28(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-(thiophene- 2-yl)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 28) Synthesis

According to Example 24, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-(thiophen-2-yl)benzo[d]oxazol-2-amine of Synthetic Example 3.

Example 29(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-methylbenzo[] Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 29)

According to Example 24, the title compound, a pale yellow solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-methylbenzo[d]oxazol-2-amine of Synthetic Example 3.

Example 30(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-fluorobenzo[d]) Synthesis of Oxazo-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 30)

According to Example 24, the title compound, a pale yellow solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-fluorobenzo[d]oxazol-2-amine of Synthetic Example 3.

Example 31(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-(4-chlorobenzene) The synthesis of (-)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 31) become

(Step 1) Synthesis of 5-(4-chlorophenyl)benzo[d]oxazol-2-amine

Following step 1 of Example 3, 4-chlorophenylboronic acid was used instead of thiophene-2-ylboronic acid to obtain the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 245.1

(Step 2) Synthesis of Compound 31 in Example

According to Example 24, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid obtained in Synthesis Example 3 and 5-(4-chlorophenyl)benzo[d]oxazol-2-amine obtained in step 1 above.

Example 32 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(di) (methylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (implementation) Synthesis of compound 32 (Example)

In step 1 of Example 24, 15 mg of (R)-tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester was added to 1.5 mL of 4N hydrochloric acid/1,4-dioxane, and stirred for 10 minutes. The solvent was then removed using an evaporator, and the mixture was azeotropically treated with toluene. The residue was dissolved in 1 mL of DMF, and 13 μL of DIPEA, 3.7 mg of (E)-4-(dimethylamino)but-2-enoate, and 8.4 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 4.2 mg of the title compound as a white solid.

Example 33 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(ethyl) (methyl)amino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide Synthesis of (Example Compound 33)

According to Example 32, (E)-4-(ethyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 34 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(di) (Ethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (implementation) Synthesis of compound 34 (Example)

According to Example 32, (E)-4-(diethylamino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 35 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(iso) propyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-carboxyl Synthesis of amines (Example compound 35)

According to Example 32, (E)-4-(isopropyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 36 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyridine) (1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide Synthesis of (Example Compound 36)

According to Example 32, (E)-4-(pyrrolidone-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 37 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(piperazolyl) ... (Pyridine-1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (actual) Synthesis of Example Compound 37

According to Example 32, (E)-4-(piperidin-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 38(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-methoxybenzene) Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 38)

According to Example 6, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-methoxybenzo[d]oxazol-2-amine of Synthetic Example 3.

Example 39(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-cyanobenzo) Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 39)

According to Example 6, the title compound, a white solid, was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-cyanobenzo[d]oxazol-2-amine of Synthetic Example 3.

Example 40(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-(2-methoxy) (Benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 40) Synthesis

According to Example 6, a pale yellow solid was obtained from (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid and 5-(2-methoxyethyl)benzo[d]oxazol-2-amine of Synthetic Example 3.

Example 41(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-phenylbenzo) Synthesis of [d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 41)

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester

20 mg of (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid from Synthesis Example 3 was suspended in 1 mL of THF. 12 mg of CDI was added at room temperature with stirring. The mixture was stirred overnight at room temperature, and 24 mg of 5-phenylbenzo[d]oxazol-2-amine was added. The mixture was then chilled, and 172 μL of a 1.0 M THF solution of lithium hexamethyldisilazane was added dropwise. After stirring for 1 hour, 30 μL of acetic acid was added to stop the reaction. The residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 12.8 mg of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 541.1

(Step 2) Synthesis of Compound 41 in Example

12.8 mg of (R)-tert-butyl-3-(4-amino-3-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester obtained in step 1 above was added to 1.5 mL of 4N hydrochloric acid/1,4-dioxane and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically reacted with 1 mL of toluene. 1 mL of chloroform and 16 μL of triethylamine were added to the residue, and the mixture was stirred under ice-cold conditions. 1.9 μL of acryloyl chloride was added to the solution and stirred for 1 hour. The reaction was stopped with saturated sodium bicarbonate aqueous solution, and the mixture was extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 3.46 mg of the title compound as a white solid.

Example 42 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-1-(1-(4-) (dimethylamino)but-2-enoyl)pyrrolid-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-carboxamide Synthesis of (Example Compound 42)

In step 1 of Example 41, 5 mg of (R)-tert-butyl-3-(4-amino-3-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane, and stirred for 30 minutes. The solvent was then removed, and the mixture was further azeotropically treated with 1 mL of toluene. The residue was dissolved in 1 mL of DMF, and 7.9 μL of DIPEA, 2.2 mg of (E)-4-(dimethylamino)but-2-enoate, and 5.18 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 3.04 mg of the title compound as a white solid.

Example 43(R)-1-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-(trifluoromethyl) The synthesis of (-)benzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example compound 43) become

(Step 1) Synthesis of (R)-tert-butyl-3-(4-amino-3-((5-(trifluoromethyl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester

32 mg of (R)-4-amino-1-(1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid from Synthesis Example 3 was suspended in 2 mL of THF. 55 mg of CDI was added at room temperature with stirring. The mixture was stirred overnight at room temperature, and 28 mg of 5-(trifluoromethyl)benzo[d]oxazol-2-amine was added. The mixture was then chilled, and 183 μL of a 1.0 M THF solution of lithium hexamethyldisilazane was added dropwise. After stirring for 1 hour, water was added, and the resulting solid was filtered off. The solid was washed with a mixture of hexane and ethyl acetate to give 35 mg of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 533.3

(Step 2) Synthesis of Compound in Example 43

In step 1 above, 8 mg of (R)-tert-butyl-3-(4-amino-3-((5-(trifluoromethyl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester was added to 500 μL of dichloromethane, followed by 200 μL of trifluoroacetic acid, and stirred for 30 minutes. The solvent was then removed, and the mixture was further azeotropically reacted with 1 mL of toluene. 2 mL of chloroform and 11 μL of triethylamine were added to the residue, and the mixture was stirred under ice-cold conditions. 1.2 μL of acryloyl chloride was added to the solution, and after stirring for 1 hour, the reaction was stopped with a saturated sodium bicarbonate aqueous solution, followed by extraction with chloroform. The organic layer was dried with sodium sulfate, and the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 1.58 mg of the title compound as a white solid.

Example 44 (R, E)-4-amino-N-(5-(trifluoromethyl)benzo[d]oxazol-2-yl)-1-(1- (4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-carboxyl Synthesis of amines (Example compound 44)

In step 1 of Example 43, 5 mg of (R)-tert-butyl-3-(4-amino-3-((5-(trifluoromethyl)benzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylic acid ester was added to 500 μL of dichloromethane, followed by 200 μL of trifluoroacetic acid, and stirred for 30 minutes. The solvent was then removed, and the mixture was further azeotropically treated with 1 mL of toluene. The residue was dissolved in 1 mL of DMF, and 6.5 μL of DIPEA, 1.9 mg of (E)-4-(dimethylamino)but-2-enoate, and 4.3 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 2.88 mg of the title compound as a white solid.

Example 45 1-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxa) Synthesis of (-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide (Example Compound 45)

(Step 1) Synthesis of tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester

240 mg of tert-butyl 3-hydroxyacetidine-1-carboxylic acid was dissolved in 2 mL of chloroform. 290 μL of triethylamine and 130 μL of methanesulfonyl chloride were added at 0 °C. After stirring for 1 hour under ice-cold conditions, chloroform and water were added to separate the organic layer. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate and water, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. 300 mg of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine-4-amine, 570 mg of potassium carbonate, and 3 mL of DMA (synthesized by the method described in International Publication No. 2007/126841) were added to the residue. The mixture was heated to 100 °C and stirred for 11 hours. After cooling to room temperature, the residue was extracted with ethyl acetate, washed with water, and dried over anhydrous magnesium sulfate. The residue was purified by amine gel chromatography (hexane/ethyl acetate = 1:1 → 0:1) to give 232 mg of the title compound as a pale yellow solid.

Physical property value: m/z[M+H] ⁺ 417.1

(Step 2) Synthesis of 4-amino-1-(1-(tert-butoxycarbonyl)acetidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid

262 mg of tert-butyl 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylic acid ester obtained in step 1 was dissolved in 10 mL of methanol and 1 mL of triethylamine. After converting to a carbon monoxide atmosphere, 51 mg of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)-dichloromethane was added, and the mixture was heated at 80 °C for 14 hours. After cooling, the solvent in the solution was removed, and 1 mL of 1,4-dioxane was added to the residue, followed by 500 μL of 5N sodium hydroxide aqueous solution. After stirring at room temperature for 3 hours, the pH was adjusted to 4 with 2N hydrochloric acid, and the mixture was ice-coldened. Water was added, and the precipitated solid was filtered and dried to obtain 42 mg of the title compound as a light brown solid.

Physical property value: m/z[M+H] ⁺ 335.2

(Step 3) Synthesis of tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)acetidine-1-carboxylic acid ester

42 mg of 4-amino-1-(1-(tert-butoxycarbonyl)acetidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxylic acid obtained in step 2 above was dissolved in 3 mL of DMF, and 24 mg of CDI was added. The mixture was stirred overnight at room temperature. Another 4 mg of CDI was added and stirred for 30 minutes. 42 mg of 5-chlorobenzo[d]oxazol-2-amine was added to the solution, and the mixture was chilled. 10 mg of sodium hydride (60%) was added. After stirring for 1 hour, the reaction was stopped with water, and the solvent was removed. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 34 mg of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 485.2

(Step 4) Synthesis of Compound 45 in Example

In step 3 above, 10 mg of tert-butyl-3-(4-amino-3-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)acetidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically reacted with 1 mL of toluene. 1 mL of chloroform and 14 μL of triethylamine were added to the residue, and the mixture was stirred under ice-cold conditions. 1.7 μL of acryloyl chloride was added to the solution and stirred for 1 hour. The reaction was stopped with saturated sodium bicarbonate solution, and the mixture was extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 0.69 mg of the title compound as a white solid.

Example 46 7-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxa) Synthesis of (-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example Compound 46)

(Step 1) Synthesis of tert-butyl 3-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid tert-butyl ester

2.3 mL of DEAD was added to 80 mL of a THF solution containing 2.00 g of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine, 1.86 g of N-Boc-3-hydroxyacetidine, and 3.75 g of triphenylphosphine. The reaction mixture was stirred for 1 hour. The reaction mixture was concentrated and washed with ethyl acetate to obtain 2.55 g of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 435.0

(Step 2) Synthesis of tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester

1.5 g of tert-butyl 3-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester obtained in step 1 above was added to 6 mL of THF and 6 mL of 28% ammonia water. The reaction solution was stirred at 100 °C for 1.5 hours using a microwave reactor. The organic layer was separated by adding chloroform and water, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation to obtain 1.5 g of the title compound as a white solid.

Physical property value: m/z[M+H] ⁺ 416.0

(Step 3) Synthesis of tert-butyl 3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester

32 mg of tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester, 20 mg of 5-chlorobenzo[d]oxazol-2-amine, and 28 μL of diazabicycloundecene obtained in step 2 above were dissolved in 1 mL of DMF. Then, 9 mg of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)-dichloromethane was added, and the mixture was stirred at 80 °C for 1.5 hours under a carbon monoxide atmosphere. The mixture was separated by chloroform and water, and the organic layer was dried with sodium sulfate. The residue after solvent removal was purified by silica gel chromatography (hexane/ethyl acetate = 1/1 → ethyl acetate/methanol = 10/1) to obtain 20 mg of the title compound as a light brown solid.

Physical property value: m/z[M+H] ⁺ 484.2

(Step 4) Synthesis of Compound 46 in Example

In step 3 above, 5 mg of tert-butyl 3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically reacted with 1 mL of toluene. 1 mL of chloroform and 14 μL of triethylamine were added to the residue, and the mixture was stirred under ice-cold conditions. 1.7 μL of acryloyl chloride was added to the solution, and the mixture was stirred for 1 hour. The reaction was stopped with a saturated sodium bicarbonate aqueous solution, and the mixture was extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 2.21 mg of the title compound as a white solid.

Example 47 (E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(dimethyl) (amino)but-2-enoyl)acryl-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example) Synthesis of compound 47)

In step 3 of Example 46, 5 mg of tert-butyl 3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)acetidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically treated with 1 mL of toluene. The residue was dissolved in 1 mL of DMF, and 14.4 μL of DIPEA, 4.1 mg of (E)-4-(dimethylamino)but-2-enoate, and 9.4 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 4.67 mg of the title compound.

Example 48(R)-7-(1-Acryloylpyrrolidine-3-yl)-4-amino-N-(5-chlorobenzo[d]) Synthesis of Oxazo-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example Compound 48)

(Step 1) Synthesis of (R)-tert-butyl 3-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester

5.00 g of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine, 19.1 g of (S)-tert-butyl-3-(methanesulfonyloxy)pyrrolidine-1-carboxylic acid ester, and 23.5 g of cesium carbonate were synthesized using the method described in International Publication No. 2005/042556. The mixture was suspended in 25 mL of acetonitrile and heated at 60 °C for 3 hours. After cooling, water and methanol were added, and the resulting solid was filtered, separated, and dried to give 5.65 g of the title compound as a light brown solid.

(Step 2) (R)-tert-butyl-3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidin-1-carboxylic acid ester

5 g of (R)-tert-butyl 3-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester obtained in step 1 above was added to 40 mL of 28% ammonia water, and the reaction solution was stirred at 100 °C for 1.5 hours using a microwave reactor. After stirring for 1 hour under ice-cold conditions, the precipitated solid was filtered and separated, washed with cold methanol, and 3.91 g of the title compound as a white solid was obtained.

(Step 3) Synthesis of (R)-tert-butyl3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester

93 mg of tert-butyl 3-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester, 110 mg of 5-chlorobenzo[d]oxazol-2-amine, and 100 μL of diazabicycloundecene obtained in step 2 above were dissolved in 2 mL of DMF. Then, 35 mg of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)-dichloromethane was added, and the mixture was stirred at 80 °C for 2.5 hours under a carbon monoxide atmosphere. The mixture was separated by chloroform and water, and the organic layer was dried with sodium sulfate. The residue after solvent removal was purified by silica gel chromatography (hexane/ethyl acetate = 1/1 → ethyl acetate/methanol = 10/1) to obtain 106 mg of the title compound as a light brown solid.

Physical property value: m/z[M+H] ⁺ 498.1

(Step 4) Synthesis of Compound 48 in Example

In step 3 above, 20 mg of (R)-tert-butyl 3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester was added to 1 mL of 4N-hydrochloric acid/1,4-dioxane and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically reacted with 1 mL of toluene. 2 mL of chloroform and 28 μL of triethylamine were added to the residue, and the mixture was stirred under ice-cold conditions. 3.2 μL of acryloyl chloride was added to the solution and stirred for 1 hour. The reaction was stopped with saturated sodium bicarbonate solution, and the mixture was extracted with chloroform. After drying the organic layer with sodium sulfate, the residue after solvent removal was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 3.52 mg of the title compound as a white solid.

Example 49 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(di) (Methylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example) Synthesis of compound 49

In step 3 of Example 48, 13 mg of (R)-tert-butyl 3-(4-amino-5-((5-chlorobenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane, and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically treated with 1 mL of toluene. The residue was dissolved in 1 mL of DMF, and 14.4 μL of DIPEA, 4.1 mg of (E)-4-(dimethylamino)but-2-enoate, and 9.6 mg of HATU were added.

After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 6.66 mg of the title compound.

Example 50 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(ethyl) (methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (actual) Synthesis of Example Compound 50

According to Example 49, (E)-4-(ethyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 51 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(di) (Ethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example) Synthesis of compound 51

According to Example 49, (E)-4-(diethylamino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 52 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(iso) propyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide Synthesis of (Example Compound 52)

According to Example 49, (E)-4-(isopropyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 53 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(pyridine) (1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (actual) Synthesis of Example Compound 53

According to Example 49, (E)-4-(pyrrolidone-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 54 (R, E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(piperazol) ... (Pyridine-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (implementation) Synthesis of compound 54 (Example)

According to Example 49, (E)-4-(piperidin-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 55(R)-7-(1-acryloylpyrrolidine-3-yl)-4-amino-N-(5-phenylbenzo) Synthesis of [d]oxazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (Example Compound 55)

(Step 1) Synthesis of (R)-tert-butyl3-(4-amino-5-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester

Following step 3 of Example 48, 5-phenylbenzo[d]oxazol-2-amine was used instead of 5-chlorobenzo[d]oxazol-2-amine to obtain the title compound as a brown solid.

Physical property value: m/z[M+H] ⁺ 540.3

(Step 2) Synthesis of Compound 55 in Example

Following step 4 of Example 48, the title compound, a white solid, was obtained using (R)-tert-butyl 3-(4-amino-5-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester obtained in step 1 above.

Example 56 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (dimethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (actual) Synthesis of Example Compound 56

In step 1 of Example 55, 13 mg of (R)-tert-butyl 3-(4-amino-5-((5-phenylbenzo[d]oxazol-2-yl)carbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrrolidine-1-carboxylic acid ester was added to 1 mL of 4N hydrochloric acid/1,4-dioxane, and stirred for 1 hour. The solvent was then removed, and the mixture was further azeotropically treated with 1 mL of toluene. The residue was dissolved in 1 mL of DMF, and 14.4 μL of DIPEA, 4.1 mg of (E)-4-(dimethylamino)but-2-enoate, and 9.6 mg of HATU were added. After stirring at room temperature for 1 hour, the solution was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (water/acetonitrile (0.1% formic acid)) to give 6.66 mg of the title compound.

Example 57 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidin-5-carboxamide Synthesis of (Example Compound 57)

According to Example 56, (E)-4-(ethyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 58 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (diethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (actual) Synthesis of Example Compound 58

According to Example 56, (E)-4-(diethylamino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 59 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidin-5-carboxyl Synthesis of amine (compound 59 in Example)

According to Example 56, (E)-4-(isopropyl(methyl)amino)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 60 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (pyrrolidine-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide Synthesis of (Example Compound 60)

According to Example 56, (E)-4-(pyrrolidine-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Example 61 (R, E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-) (piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (actual) Synthesis of Example Compound 61)

According to Example 56, (E)-4-(piperidin-1-yl)but-2-enoate salt was used instead of (E)-4-(dimethylamino)but-2-enoate salt to obtain the title compound as a white solid.

Comparative Example 1 (R)-1-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine Synthesis of (piperidin-1-yl)prop-2-en-1-one (Comparative Compound 1, PCI-32765)

The title compound was synthesized according to the method described in International Publication No. 2008/121742, yielding a white solid.

The structural formulas and physical properties of compounds 1 to 61 of Examples and compound 1 of Comparative Example are shown in Tables 2 to 14 below.

[Table 2]

[Table 3]

[Table 4]

[Table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

[Table 10]

[Table 11]

[Table 12]

[Table 13]

[Table 14]

Experimental Example 1: BTK inhibitory activity (in vitro) Determination of (in vitro)

In the setting of the in vitro BTK kinase inhibitory activity assay, the price list of PerkinElmer's LabChip (registered trademark) series of reagents and consumables listed FL-Peptide 2 as the substrate peptide for the BTK kinase activity assay; therefore, FL-Peptide 2 was used as the substrate. The purified recombinant human BTK protein used in the experiment was purchased from Carna Biosciences.

In the assay of the inhibitory activity of the compound, the compound of the present invention was first serially diluted with DMSO. Next, BTK protein, substrate peptide (final concentration 1 μM), magnesium chloride (final concentration 10 mM), ATP (final concentration 45 μM), and a DMSO solution of the compound of the present invention (final concentration 5%) were added to a kinase reaction buffer (20 mM HEPES (pH 7.5), 2 mM dithiothreitol, 0.01% Triton X-100), and the reaction was incubated at 25°C for 40 minutes. EDTA was then added to bring the final concentration to 30 mM, thereby stopping the reaction. Finally, the unphosphorylated substrate peptide (S) and phosphorylated peptide (P) were separated and detected by microchannel capillary electrophoresis using a LabChip EZ Reader II (PerkinElmer). The amount of phosphorylation reaction was determined from the peak heights of S and P. The concentration of the compound that could inhibit 50% of the phosphorylation reaction was defined as the IC50 value (nM), and is shown in Tables 15 to 17 below.

[Table 15]

Example number	IC50 value of BTK inhibitory activity (nM)
		1	0.415
2	0.464
		3	0.443
4	0.888
		5	1.253
6	0.738
		7	0.457
8	1.266
		9	1.37
10	2.384
		11	2.143
12	0.433
		13	0.813
14	14.141
		15	0.786
16	0.733
		17	0.811
18	0.788
		19	0.69
20	0.801
		twenty one	0.777
twenty two	14.209
		twenty three	1.583
twenty four	0.591
		25	1.166
26	2.788

[Table 16]

Example number	IC50 value of BTK inhibitory activity (nM)
		27	1.433
28	0.559
		29	0.485
30	0.566
		31	1.671
32	0.634
		33	0.887
34	0.79
		35	0.792
36	0.867
		37	0.786
38	0.888
		39	1.12
40	2.087
		41	0.442
42	0.771
		43	0.546
44	0.877
		45	1.249
46	3.272
		47	7.345
48	0.836
		49	1.529
50	1.407
		51	1.48
52	1.195

[Table 17]

Example number	IC50 value of BTK inhibitory activity (nM)
		53	1.675
54	1.436
		55	0.799
56	1.337
		57	1.507
58	1.844
		59	1.507
60	1.88
		61	2.341

The experimental results show that the compound of the present invention has BTK inhibitory activity in the in vitro.

BTK inhibition selectivity compared with EGFR kinase inhibitory activity in Experiment Example 2 (in vitro)

1) BTK inhibitory activity assay

The BTK inhibitory activity was measured using the same procedure as in Experimental Example 1.

2) EGFR inhibitory activity assay

In the setting of the in vitro EGFR kinase inhibitory activity assay, the price list of PerkinElmer's LabChip (registered trademark) series of reagents and consumables listed FL-Peptide 22 as the substrate peptide in the EGFR kinase activity assay. Therefore, based on its amino acid sequence, a biotinylated peptide (biotin-EEPLYWSFPAKKK) was prepared. The purified recombinant human EGFR protein used in the experiment was purchased from Carna Biosciences.

In the assay of the inhibitory activity of the compound, the compound of the present invention was first serially diluted with DMSO. Then, EGFR protein, substrate peptide (final concentration 250 nM), magnesium chloride (final concentration 10 mM), manganese chloride (final concentration 10 mM), ATP (final concentration 1.5 μM), and a DMSO solution of the compound of the present invention (final concentration 2.5%) were added to a kinase reaction buffer (20 mM HEPES (pH 7.5), 2 mM dithiothreitol, 0.01% Triton X-100), and the kinase reaction was carried out at 25°C for 120 minutes. EDTA was added to bring the final concentration to 24 mM, thereby stopping the reaction. Then, a detection solution containing Eu-tagged antiphosphotyrosine antibody PT66 (PerkinElmer) and SureLight APC-SA (PerkinElmer) was added, and the mixture was incubated at room temperature for at least 2 hours. Finally, the fluorescence intensity was measured using a PHERAstar FS (BMG LABTECH) at two wavelengths, 620 nm and 665 nm, when irradiated with excitation light at a wavelength of 337 nm. The phosphorylation reaction intensity was determined based on the ratio of fluorescence intensity at the two wavelengths, and the concentration of the compound that could inhibit the phosphorylation reaction by 50% was defined as the IC50 value (nM).

3) BTK inhibition selectivity

Based on the results obtained in 1) and 2) above, the “EGFR inhibitory activity IC50 value (nM) / BTK inhibitory activity IC50 value (nM)” is calculated to confirm the BTK inhibitory selectivity of the tested compound.

[Table 18]

[Table 19]

The experimental results clearly demonstrate that, in vitro, the BTK inhibitory selectivity of the compound of the present invention relative to EGFR kinase is more than 7.5 times that of comparative compound 1, indicating that the compound of the present invention exhibits excellent BTK inhibitory selectivity. This result suggests that the compound of the present invention may reduce side effects compared to known BTK inhibitors.

Experimental Example 3: Assay on the inhibitory activity of BTK and EGFR-expressing cell lines on cell proliferation (in) (in vitro) and its selection Sexual comparison

TMD8 cells, a diffuse large cell B-cell lymphoma cell line expressing BTK, were suspended in RPMI 1640 medium (Life Technologies) containing 10% fetal bovine serum. A431 cells, a highly activated human epithelial cancer cell line overexpressing EGFR, were suspended in DMEM high-glucose medium (Life Technologies) containing 10% fetal bovine serum. The cell suspensions were seeded into the wells of 384-well flat-bottom microplates and cultured at 37°C for 1 day in a culturer containing 5% CO2. The compounds of the present invention and Comparative Example 1 were dissolved in DMSO, and the test compounds were diluted 500 times to their final concentration using DMSO. The DMSO solution of the test compounds was diluted with the medium used for the cell suspensions and added to the wells of the cell culture plates to achieve a final DMSO concentration of 0.2%. The cells were then cultured at 37°C for another 3 days in a culturer containing 5% CO2. Cell counts were performed before the addition of the compound and 3 days after incubation in the presence of the compound, using a Celliter Glo (Promega) according to Promega's recommended protocol. The proliferation inhibition rate was calculated using the following formula, and the concentration of the tested compound (GI50 (nM)) at which 50% inhibition was achieved was determined.

Proliferation inhibition rate (%) = (C - T) / (C - CO) × 100 T: Luminescence intensity of the pore after adding the tested compound

C: Luminescence intensity of pores without the tested compound added

C0: Luminescence intensity of the pore measured before the addition of the compound being tested.

Comparing the cell proliferation inhibitory activities of A431 cells (dependent on EGFR proliferation signaling) and TMD8 cells (dependent on BTK proliferation signaling) allows for the evaluation of the effects on various kinases at the cellular level. Specifically, the ratio of "A431 cell proliferation inhibition rate to TMD8 cell proliferation inhibition rate" is calculated; a higher value indicates higher BTK selectivity relative to EGFR in the cells. Tables 20 and 21 represent the values of "A431 cell proliferation inhibition rate to TMD8 cell proliferation inhibition rate".

[Table 20]

[Table 21]

The experimental results show that the BTK inhibitory selectivity of the compound of the present invention relative to EGFR kinase in inhibiting cell proliferation in vitro is more than 8.5 times that of comparative compound 1. Therefore, the compound of the present invention exhibits excellent BTK inhibitory selectivity not only against kinases but also in cells. This result suggests that the compound of the present invention may reduce side effects compared to existing BTK inhibitors.

Experiment 4: Inhibition of B cell activation using Ramos cells

Ramos cell line, derived from human B-cell lymphoma, was suspended in RPMI 1640 medium and seeded at a concentration of 2.0 × ^10⁶ (cells/well) in culture plates and cultured at 37°C in a _CO₂ incubator (SANYO) for 12 hours. Comparative Example Compound 1, Example Compound 12, and Example Compound 13 were serially diluted with DMSO and added to the cell-seeded plates, and cultured in a _CO₂ incubator for 1 hour. Afterward, the cells were stimulated for 10 minutes with Goat F(ab')₂ anti-human IgM antibody-UNLB (Southern Biotech). Cells were then recovered, and cell extract (NP-40, Invitrogen) containing 50 μL of a combination of 1× protease inhibitor (Roche) and 1× phosphatase inhibitor (cocktail inhibitor) (Sigma) was added to the cell clusters. The cells were incubated on ice for 10 minutes. Protein levels in the recovered cell extract were quantified using a DC protein assay (Biorad). 20 μg of protein was added per lane to a Criterion TGX™ (Biorad) electrophoresis apparatus, followed by Western blotting using a Trans-Blot ^™ Turbo ^™ (Biorad). Immunoblotting was then performed using a LAS4000 (GE) system with BTK phosphor (pY223) antibody (EPITOMICS) and BTK antibody (Abcam). Healthcare, Inc. detected phosphorylated BTK protein and BTK protein. Then, based on the luminescence intensity of the detected proteins, the ratio of phosphorylated BTK protein to BTK protein was calculated, and the concentration of the compound that could inhibit 50% of phosphorylated BTK protein was defined as the IC50 value (nM). Table 22 shows the BTK phosphorylation inhibition concentration (IC50, (nM)).

As shown in Table 22, Comparative Compound 1 exhibited inhibition of BTK phosphorylation at 0.70 (nM), while the compound of the present invention exhibited equal or greater inhibition of BTK phosphorylation. The compound of the present invention is able to inhibit B cell activation signals mediated by B cell receptor stimulation.

Rituxan, which targets B cells, is known to be effective against autoantibody-induced autoimmune diseases, including rheumatoid arthritis (non-patent literature: Rastetter et al., Annu Rev Med, 55, 2004).

Based on this experiment, it was confirmed that the compound of the present invention can inhibit the activation of B cells mediated by B cell receptor stimulation, and can be considered to have excellent efficacy against autoimmune diseases involving B cells that produce autoantibodies.

[Table 22]

Example number	Inhibitory concentration of phosphorylated BTK (IC50, (nM))
		12	0.18
13	0.25
		Comparative compound 1	0.70

Experimental Example 5: Inhibition of Allergic Reactions Using RBL-2H3 Cells

Rat basophilic leukemia cells (RBL-2H3) were suspended in MEM medium containing 10% FBS and seeded into culture plates. The cells were cultured at 37°C in a _CO2 incubator (SANYO) for 12 hours. After removing the culture supernatant and washing the cells, anti-DNP-mouse IgE solution (Alpha Diagnostic Inc.) was added, and the cells were cultured in a _CO2 incubator for 1 hour. After removing the culture supernatant and washing the cells, MEM medium was added. Comparative Example Compound 1, Example Compound 1, Example Compound 12, and Example Compound 13 were serially diluted with DMSO and added to the cell-seeded plates. The plates were then cultured in a _CO2 incubator for 30 minutes.

DNP-BSA (LSL) solution was added, and the mixture was incubated in a _CO2 incubator for 15 minutes. The culture supernatant was recovered, and the reaction was carried out using the _PGD2 -MOX EIA kit (Cayman Chemical) according to the accompanying protocol. The absorbance was measured using a SUNRISE RAINBOW THERMO (TECAM). Based on the measured absorbance, the amount of _PGD2 in the culture supernatant was calculated and compared with a reference. The concentration of the compound that inhibited _PGD2 production by 50% was defined as the IC50 value (nM). Table 23 shows the _PGD2 production inhibition concentration (IC50, (nM)).

According to Table 23, Comparative Compound 1 exhibited PGD ₂ inhibition at approximately 350 (nM), while the compound of the present invention exhibited a much stronger PGD ₂ inhibition ability than the comparative compound.

IgE antibodies bind to FCε receptors on mast cells. When these antibodies cross-link with antigens, mast cells are activated, releasing or secreting various neurotransmitters (histamine, _PGD2 , or leukotrienes). These are known to be closely associated with the pathogenesis of allergic diseases such as bronchial asthma or allergic rhinitis (Non-patent literature: Ellmeier W., et al., FEBS Journal., 278, 2011).

It can be confirmed that the compounds of the present invention control the release or secretion of chemical neurotransmitters in mast cells downstream of the FCε receptor, and therefore can be considered to exhibit excellent efficacy against allergic diseases.

[Table 23]

Experiment 6: Mouse collagen-induced arthritis model (preventive effect)

This experiment was conducted according to the methods described in non-patent literature (Brand DD, et al., Nat Protoc. 2007; 2, 1269-1275, Xu D. et al., JPET, 2012 Apr; 341(1): 90-103). Seven-week-old male/DBA/1 mice (CHARLES RIVER LABORATORIES JAPAN, INC.) were injected intradermally in the back with 100 μL/body of a mixture of 4 mg/mL bovine type 2 collagen solution (Collagen Research Center) and Freund's complete adjuvant (DIFCO). Twenty-one days later, a booster immunization was performed by intradermal injection of 100 μL/body of a mixture of 4 mg/mL bovine type 2 collagen solution (Collagen Research Center) and Freund's complete adjuvant (DIFCO). Including the day of additional immunization (day 0), the drug was administered for 21 consecutive days, with the control (Vehicle), Example Compound 12, Example Compound 13, or Comparative Compound 1 given orally once daily. On days 0, 4, 7, 10, 14, 17, and 21, arthritis symptoms were visually assessed (0: no change, 1: one toe swollen, 2: two or more toes swollen, 3: dorsum of the foot swollen, 4: all toes swollen and swelling extending to the wrist). The total number of points for each limb was used as the individual's score (maximum 16 points), and the effects of each treatment group in the same model were compared. The results are shown in Figure 1.

As confirmed by Figure 1, the compound of the present invention, compared with comparative compound 1, can completely inhibit the increase in arthritis scores after additional immunization, demonstrating excellent preventive effects against the onset of rheumatoid arthritis. Furthermore, at the dosage of the compound used in this experiment, no significant weight loss, skin disorders including hair loss, or other toxicities were observed.

Experiment 7: Mouse collagen-induced arthritis model (therapeutic effect)

This experiment was conducted according to the methods described in non-patent literature (Brand DD, et al., Nat Protoc. 2007; 2, 1269-1275, Xu D. et al., JPET, 2012 Apr; 341(1): 90-103). Seven-week-old male/DBA/1 mice (CHARLES RIVER LABORATORIES JAPAN, INC.) were injected intradermally in the back with 100 μL/body of a mixture of 4 mg/mL bovine type 2 collagen solution (Collagen Research Center) and Freund's complete adjuvant (DIFCO). A booster immunization was performed 21 days later by intradermal injection of 100 μL/body of a mixture of 4 mg/mL bovine type 2 collagen solution (Collagen Research Center) and Freund's complete adjuvant (DIFCO). The administration start date (day 0) was designated as day 6 after the booster immunization. The control, compound 13 of example, or compound 1 of comparative example were administered orally once daily for 15 days. Symptoms of arthritis were visually assessed on days 0, 4, 7, 11, and 14 (0: no change, 1: swelling of one toe, 2: swelling of two or more toes, 3: swelling of the dorsum of the foot, 4: swelling of all toes extending to the wrist). The total score for each limb was recorded as a point total (maximum 16 points). The results are shown in Figure 2.

As can be confirmed from Figure 2, Comparative Compound 1 was able to delay the further increase in the rising arthritis score, but the compound of the present invention was able to reduce the arthritis score. The compound of the present invention has excellent therapeutic effects on pre-existing rheumatoid arthritis. Furthermore, at the dosage of the compound used in this experiment, no significant weight loss, skin disorders including hair loss, or other toxicities were observed.

Experimental Example 8: Mouse Collagen-Induced Arthritis Model (Therapeutic Effect)

The same tests as in Test Example 7 were performed using a control, Example Compound 12, and Comparative Compound 1. The results are shown in Figure 3.

As can be confirmed from Figure 3, Comparative Compound 1 delayed the further increase in arthritis scores, but the compound of the present invention significantly reduced arthritis scores. The compound of the present invention exhibits excellent therapeutic effects on pre-existing rheumatoid arthritis. Furthermore, at the dosages of the compound used in this experiment, no significant weight loss, skin disorders including hair loss, or other toxicities were observed.

Experimental Example 9: Mouse Antigen-Induced Dermatitis Model

A type I allergy model (dermatitis model) with skin symptoms was established using TNP-IgE mice (CLEA Japan, Inc.). These mice are genetically modified BALB/c mice that routinely produce specific IgE from the hapten TNP (tinitrophenol), a commonly used allergen (antigen). Allergic reactions can be induced simply by administration of the allergen without immunization. The auricular skin of both ears of these mice was coated with 10 μL/ear of a 0.025% acetone solution of picryl chloride (NACALAITESQUE, INC.), the allergen. The thickness of the auricle was measured 2 hours after antigen application using a dial-type thickness gauge (Ozaki Corporation, PEACOCK G-2M). The average thickness values of both ears (with the initial value before allergen application set as 0 mm) were taken as the individual value. The control, comparative compound 1, example compound 1, example compound 6, example compound 12, or example compound 13 were orally administered 30 minutes before allergen application.

The auricular swelling inhibition rate (%) was calculated using the following formula. The results are shown in Table 24.

Auricular swelling inhibition rate (%) = (1 - (average auricular thickness in the compound-treated group) / (average auricular thickness in the control group)) × 100

[Table 24]

According to Table 24, it can be confirmed that, compared with Comparative Compound 1, Compound 1, Compound 6, Compound 12 and Compound 13 of Examples can inhibit auricular swelling induced by the application of equal or greater amounts of allergens at low doses. The compounds of the present invention show excellent efficacy against atopic dermatitis.

Experimental Example 10: Guinea Pig Antigen-Induced Rhinitis Model

A type I allergy model (rhinitis model) with nasal congestion was established using guinea pigs (Japan SLC, Inc.). The sensitization day was designated as day 0. Sensitization was achieved by subcutaneously administering 1 mL of 1 mg/mL ovalbumin saline solution (OVA solution) per body using a 1 mL syringe with a 23G needle. On days 7, 14, and 21, 20 μL of 10 mg/mL OVA solution was administered intranasally to each of the two nasal cavities to induce an allergic reaction. Two hours before antigen induction on day 21, a single oral administration of the control or compound 13 from example was administered. Nasal resistance was measured on day 20 as the pre-value for each individual, and nasal resistance was measured 15 minutes and 240 minutes after antigen induction on day 21.

The nasal cavity resistance value (rate of change, %) was calculated using the following formula. The results are shown in Figure 4.

Nasal resistance value (rate of change, %) = ((nasal resistance value of the control group or the compound of the example group (15 minutes or 240 minutes, day 21)) / (nasal resistance value of the control group or the compound of the example group (pre, day 20)) - 1) × 100

According to Figure 4, compared with the control group that showed an increase in nasal resistance due to antigen induction, the use of compound 13 of Example showed that the increase in nasal resistance was inhibited in both the immediate and delayed phases, demonstrating dose-responsiveness. Therefore, it can be confirmed that the compound of the present invention exhibits excellent efficacy against allergic rhinitis and hay fever.

Experimental Example 11: Effects of repeated administration of the compound of the present invention on body weight, etc., in SD rats (in vivo) vivo

The effects of repeated two-week administration of Comparative Example Compound 1 and the compound of the present invention on the weight gain of SD rats were compared with those of the solvent-administered group. To ensure that the mean weight of each group was approximately uniform, rats were randomly assigned to groups of four (day 1) as follows.

Group 1: Comparative compound 1 (280 mg/kg) was administered orally once daily.

Group 2: Compound 12 (750 mg/kg) from Example 1 was administered orally once daily.

Group 3: Compound 13 (750 mg/kg) from Example 1 was administered orally once daily.

As an indicator of systemic toxicity caused by the administration of the compound, body weight change (BWC) is used. BWC is calculated according to the following formula.

BWC (%) = ([(Rats' body weight on day 14 after drug administration) - (Rats' body weight at the time of grouping)] / (Rats' body weight at the time of grouping)) × 100

The relative weight change rate of each compound administration group was calculated using the following formula, with the BWC of the solvent administration group set to 1, and is shown in Table 25.

Relative body weight change rate (%) = (BWC of the compound-treated group) / (BWC of the solvent-treated group) × 100

[Table 25]

Group	Dosage (mg/kg)	Relative body weight change rate (%)
			Group 1	280	30.9
Group 2	750	91.2
			Group 3	750	79.1

As a result, compared with the solvent-administered group, the weight gain of rats in group 1, which was the group treated with compound 1 (comparative example), was only slightly increased. In contrast, groups 2 and 3, which were the groups treated with the compounds of the present invention, had almost no effect on the weight gain of rats despite being administered at doses more than 2.5 times that of comparative example compound 1 (approximately 5 times the dose of AUC _0-24 (μM:hr)). Furthermore, in group 1, individuals were found to have soft stools, while none were found in groups 2 and 3. Therefore, the compounds of the present invention exhibit excellent effects, such as low side effects even at much higher exposure levels than comparative example compound 1.

The results above show that the compound group of the present invention has excellent properties (profile) with reduced toxicity compared with comparative example compound 1.

Experimental Example 12: Mouse model of systemic lupus erythematosus

MRL/lpr mice (Japan SLC, Inc.) are commonly used as mouse models of autoimmune diseases that naturally manifest symptoms similar to human systemic lupus erythematosus (SLE), and were therefore used for efficacy evaluation in this study. MRL/lpr mice were grouped based on swollen lymph nodes in the mandible and axilla at 14 weeks of age. Forty-two days after the grouping date, mice were orally administered once daily a suspension of either the control or Example 13. Peripheral blood urea nitrogen and anti-dsDNA antibody concentrations were measured on the final administration date. The results are shown in Figure 5.

As confirmed by Figure 5, compared with the control drug, compound 13 of the examples was able to reduce blood urea nitrogen, an elevated indicator of renal function, and anti-dsDNA antibody, an autoantibody. The compounds of the present invention showed excellent efficacy against pre-existing systemic lupus erythematosus. Furthermore, no significant weight loss was observed at the dosage of the compounds used in this experiment.

Prophylactic agents and/or therapeutic agents for immune diseases

Technical Field

Background Technology

Existing technical documents

Patent documents

Patent Document 1: International Publication No. 2011/090760

Patent Document 2: International Publication No. 2009/158571

Non-patent literature

Non-patent literature 3: Ellmeier W., et al., FEBS Journal., 278, 2011

Non-patent literature 4: Rommel C., et al., Nature Reviews Immunology, 7, 2007

Non-patent literature 6: Lacouture ME., Nature Reviews Cancer, 6, 2006

Summary of the Invention

The problem that the invention aims to solve

Methods for solving problems

Y represents -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z independently represent N or CH, respectively;

n represents an integer from 0 to 2;

R ₁ represents an amino group that can have substituents;

Invention Effects

Attached Figure Description

Figure 4 illustrates the effect of guinea pig antigen-induced rhinitis model.

Figure 5 shows the effect on a mouse model of systemic lupus erythematosus.

Detailed Implementation

In general formula (I), Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 .

In general formula (I), n is preferably 0.

X is a nitrogen-containing C3-C10 heterocyclic alkyl group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

(1) When Z is N and W is N.

X is 1,3-piperidinyl,

Y represents vinyl.

(2) When Z is CH and W is N.

X is 1,3-pyridinealkyl or 1,3-piperidineyl.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

(3) When Z is CH and W is CH.

X is 1,3-adiazine-1,3-pyrrolidine-1,3-diphenylene ...

Y is -C( _R4 ) = C( _R5 )( _R6 ).

n is 0;

_R1 is an amino group;

(1) When Z is N and W is N.

Y represents vinyl.

(2) When Z is CH and W is N.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

(3) When Z is CH and W is CH.

Y is -C( _R4 ) = C( _R5 )( _R6 ).

n is 0;

_R1 is an amino group;

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Manufacturing Method 1

[In the formula, Z, _R2 , and _R3 have the same meaning as above.]

Manufacturing Method 2

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

Manufacturing method 3

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

Manufacturing method 4

Manufacturing Method 5

(Process 8, Process 9)

(Process 10)

Examples of solvents include water, propylene glycol, and physiological saline.

Examples of analgesics include procaine hydrochloride and lidocaine.

Example

Column: YMC-Triart C18, 2.0 x 50 mm, 1.9 μm, manufactured by YMC Corporation.

MS test: ESI positive

UV detection: 254 and 210nm

Column flow rate: 0.5 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 1 μL

Gradient (Table 1)

[Table 1]

Time (minutes)	water	Acetonitrile
			0	95	5
0.1	95	5
			2.1	5	95
3.0	Stop

UV detection: 254nm

MS test: ESI positive

Column flow rate: 25 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 0.1–0.5 mL

The meanings of the abbreviation symbols are as follows.

s: single peak

d: double peak

t: triple peak

q: Four Peaks

dd: Double double peak

dt: double and triple peaks

td: triple and double peaks

tt: Double Triple Peak

ddd: double double double peak

ddt: doublet doublet triplet peak

dtd: double triple double peak

tdd: triple double double peak

m: multiple peaks

br: broad peak

brs: broad single peak

CDI: Carbonyldiimidazole

DMSO- _d6 : Deuterated dimethyl sulfoxide

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

THF: Tetrahydrofuran

DMF: N,N-Dimethylformamide

DMA: N,N-dimethylacetamide

NMP: 1-Methyl-2-pyrrolidone

DMSO: Dimethyl sulfoxide

TFA: Trifluoroacetic acid

WSC: 1-(3-dimethylaminopropyl)-3- and carbodiimide hydrochloride

HOBt: 1-Hydroxybenzotriazole hydrate

DIAD: Diisopropyl azodicarbonate

TBAF: Tetrabutylammonium fluoride

DIPEA: Diisopropylethylamine

Boc: tert-Butoxycarbonyl

Boc ₂ O: Ditert-butyl dicarbonate

DMAP: Dimethylaminopyridine

Physical property value: m/z[M+H] ⁺ 446.2

Physical property value: m/z[M+H] ⁺ 363.1

Physical property value: m/z[M+H] ⁺ 266.1

Physical property value: m/z[M+H] ⁺ 431.0

Physical property value: m/z[M+H] ⁺ 349.1

Synthesis Example 4: Synthesis of 5-cyanobenzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 161.1

Physical property value: m/z[M+H] ⁺ 513.2

(Step 2) Synthesis of Compound 1 in Example

(Step 1) Synthesis of 5-(thiophene-2-yl)benzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 216.8

Physical property value: m/z[M+H] ⁺ 561.3

(Step 3) Synthesis of Compound 3 in Example

Physical property value: m/z[M+H] ⁺ 505.3

(Step 2) Synthesis of Compound 6 in Example

Physical property value: m/z[M+H] ⁺ 497.2

Physical property value: m/z[M+H] ⁺ 398.0

(Step 3) Synthesis of Compound 12 in Example

Physical property value: m/z[M+H] ⁺ 479.3

(Step 2) Synthesis of Compound 13 in Example

Physical property value: m/z[M+H] ⁺ 318.1

(Step 2) Synthesis of Compound 23 in Example

Physical property value: m/z[M+H] ⁺ 499.1

(Step 2) Synthesis of Compound 24 in Example

(Step 1) Synthesis of 5-(4-chlorophenyl)benzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 245.1

(Step 2) Synthesis of Compound 31 in Example

Physical property value: m/z[M+H] ⁺ 541.1

(Step 2) Synthesis of Compound 41 in Example

Physical property value: m/z[M+H] ⁺ 533.3

(Step 2) Synthesis of Compound in Example 43

Physical property value: m/z[M+H] ⁺ 417.1

Physical property value: m/z[M+H] ⁺ 335.2

Physical property value: m/z[M+H] ⁺ 485.2

(Step 4) Synthesis of Compound 45 in Example

Physical property value: m/z[M+H] ⁺ 435.0

Physical property value: m/z[M+H] ⁺ 416.0

Physical property value: m/z[M+H] ⁺ 484.2

(Step 4) Synthesis of Compound 46 in Example

Physical property value: m/z[M+H] ⁺ 498.1

(Step 4) Synthesis of Compound 48 in Example

Physical property value: m/z[M+H] ⁺ 540.3

(Step 2) Synthesis of Compound 55 in Example

[Table 2]

[Table 3]

[Table 4]

[Table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

[Table 10]

[Table 11]

[Table 12]

[Table 13]

[Table 14]

[Table 15]

[Table 16]

[Table 17]

1) BTK inhibitory activity assay

2) EGFR inhibitory activity assay

3) BTK inhibition selectivity

[Table 18]

[Table 19]

C: Luminescence intensity of pores without the tested compound added

[Table 20]

[Table 21]

Experiment 4: Inhibition of B cell activation using Ramos cells

[Table 22]

Experimental Example 5: Inhibition of Allergic Reactions Using RBL-2H3 Cells

[Table 23]

Experiment 6: Mouse collagen-induced arthritis model (preventive effect)

Experiment 7: Mouse collagen-induced arthritis model (therapeutic effect)

Experimental Example 9: Mouse Antigen-Induced Dermatitis Model

[Table 24]

Experimental Example 10: Guinea Pig Antigen-Induced Rhinitis Model

Group 1: Comparative compound 1 (280 mg/kg) was administered orally once daily.

[Table 25]

Experimental Example 12: Mouse model of systemic lupus erythematosus

Technical Field

Background Technology

Existing technical documents

Patent documents

Patent Document 1: International Publication No. 2011/090760

Patent Document 2: International Publication No. 2009/158571

Non-patent literature

Non-patent literature 3: Ellmeier W., et al., FEBS Journal., 278, 2011

Non-patent literature 4: Rommel C., et al., Nature Reviews Immunology, 7, 2007

Non-patent literature 6: Lacouture ME., Nature Reviews Cancer, 6, 2006

Summary of the Invention

The problem that the invention aims to solve

Methods for solving problems

Y represents -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z independently represent N or CH, respectively;

n represents an integer from 0 to 2;

R ₁ represents an amino group that can have substituents;

Invention Effects

Attached Figure Description

Figure 4 illustrates the effect of guinea pig antigen-induced rhinitis model.

Figure 5 shows the effect on a mouse model of systemic lupus erythematosus.

Detailed Implementation

In general formula (I), Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 .

In general formula (I), n is preferably 0.

X is a nitrogen-containing C3-C10 heterocyclic alkyl group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are independently N or CH;

n is 0;

_R1 is an amino group;

X is acrylonitrile, pyrrolidinyl, or piperidinyl;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z are either N or CH;

n is 0;

_R1 is an amino group;

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

X is 1,3-adiazinyl, 1,3-pyrrolidine, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

(1) When Z is N and W is N.

X is 1,3-piperidinyl,

Y represents vinyl.

(2) When Z is CH and W is N.

X is 1,3-pyridinealkyl or 1,3-piperidineyl.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

(3) When Z is CH and W is CH.

X is 1,3-adiazine-1,3-pyrrolidine-1,3-diphenylene ...

Y is -C( _R4 ) = C( _R5 )( _R6 ).

n is 0;

_R1 is an amino group;

(1) When Z is N and W is N.

Y represents vinyl.

(2) When Z is CH and W is N.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

(3) When Z is CH and W is CH.

Y is -C( _R4 ) = C( _R5 )( _R6 ).

n is 0;

_R1 is an amino group;

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

Manufacturing Method 1

[In the formula, Z, _R2 , and _R3 have the same meaning as above.]

Manufacturing Method 2

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

Manufacturing method 3

The reaction time is 0.1 to 100 hours, preferably 0.5 to 24 hours.

Manufacturing method 4

Manufacturing Method 5

(Process 8, Process 9)

(Process 10)

Examples of solvents include water, propylene glycol, and physiological saline.

Examples of analgesics include procaine hydrochloride and lidocaine.

Example

Column: YMC-Triart C18, 2.0 x 50 mm, 1.9 μm, manufactured by YMC Corporation.

MS test: ESI positive

UV detection: 254 and 210nm

Column flow rate: 0.5 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 1 μL

Gradient (Table 1)

[Table 1]

Time (minutes)	water	Acetonitrile
			0	95	5
0.1	95	5
			2.1	5	95
3.0	Stop

UV detection: 254nm

MS test: ESI positive

Column flow rate: 25 mL/min

Mobile phase: Water/acetonitrile (0.1% formic acid)

Injection volume: 0.1–0.5 mL

The meanings of the abbreviation symbols are as follows.

s: single peak

d: double peak

t: triple peak

q: Four Peaks

dd: Double double peak

dt: double and triple peaks

td: triple and double peaks

tt: Double Triple Peak

ddd: double double double peak

ddt: doublet doublet triplet peak

dtd: double triple double peak

tdd: triple double double peak

m: multiple peaks

br: broad peak

brs: broad single peak

CDI: Carbonyldiimidazole

DMSO- _d6 : Deuterated dimethyl sulfoxide

CDCl ₃ : Deuterated chloroform

_CD3 OD: Deuterated methanol

THF: Tetrahydrofuran

DMF: N,N-Dimethylformamide

DMA: N,N-dimethylacetamide

NMP: 1-Methyl-2-pyrrolidone

DMSO: Dimethyl sulfoxide

TFA: Trifluoroacetic acid

WSC: 1-(3-dimethylaminopropyl)-3- and carbodiimide hydrochloride

HOBt: 1-Hydroxybenzotriazole hydrate

DIAD: Diisopropyl azodicarbonate

TBAF: Tetrabutylammonium fluoride

DIPEA: Diisopropylethylamine

Boc: tert-Butoxycarbonyl

Boc ₂ O: Ditert-butyl dicarbonate

DMAP: Dimethylaminopyridine

Physical property value: m/z[M+H] ⁺ 446.2

Physical property value: m/z[M+H] ⁺ 363.1

Physical property value: m/z[M+H] ⁺ 266.1

Physical property value: m/z[M+H] ⁺ 431.0

Physical property value: m/z[M+H] ⁺ 349.1

Synthesis Example 4: Synthesis of 5-cyanobenzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 161.1

Physical property value: m/z[M+H] ⁺ 513.2

(Step 2) Synthesis of Compound 1 in Example

(Step 1) Synthesis of 5-(thiophene-2-yl)benzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 216.8

Physical property value: m/z[M+H] ⁺ 561.3

(Step 3) Synthesis of Compound 3 in Example

Physical property value: m/z[M+H] ⁺ 505.3

(Step 2) Synthesis of Compound 6 in Example

Physical property value: m/z[M+H] ⁺ 497.2

Physical property value: m/z[M+H] ⁺ 398.0

(Step 3) Synthesis of Compound 12 in Example

Physical property value: m/z[M+H] ⁺ 479.3

(Step 2) Synthesis of Compound 13 in Example

Physical property value: m/z[M+H] ⁺ 318.1

(Step 2) Synthesis of Compound 23 in Example

Physical property value: m/z[M+H] ⁺ 499.1

(Step 2) Synthesis of Compound 24 in Example

(Step 1) Synthesis of 5-(4-chlorophenyl)benzo[d]oxazol-2-amine

Physical property value: m/z[M+H] ⁺ 245.1

(Step 2) Synthesis of Compound 31 in Example

Physical property value: m/z[M+H] ⁺ 541.1

(Step 2) Synthesis of Compound 41 in Example

Physical property value: m/z[M+H] ⁺ 533.3

(Step 2) Synthesis of Compound in Example 43

Physical property value: m/z[M+H] ⁺ 417.1

Physical property value: m/z[M+H] ⁺ 335.2

Physical property value: m/z[M+H] ⁺ 485.2

(Step 4) Synthesis of Compound 45 in Example

Physical property value: m/z[M+H] ⁺ 435.0

Physical property value: m/z[M+H] ⁺ 416.0

Physical property value: m/z[M+H] ⁺ 484.2

(Step 4) Synthesis of Compound 46 in Example

Physical property value: m/z[M+H] ⁺ 498.1

(Step 4) Synthesis of Compound 48 in Example

Physical property value: m/z[M+H] ⁺ 540.3

(Step 2) Synthesis of Compound 55 in Example

[Table 2]

[Table 3]

[Table 4]

[Table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

[Table 10]

[Table 11]

[Table 12]

[Table 13]

[Table 14]

[Table 15]

[Table 16]

[Table 17]

1) BTK inhibitory activity assay

2) EGFR inhibitory activity assay

3) BTK inhibition selectivity

[Table 18]

[Table 19]

C: Luminescence intensity of pores without the tested compound added

[Table 20]

[Table 21]

Experiment 4: Inhibition of B cell activation using Ramos cells

[Table 22]

Experimental Example 5: Inhibition of Allergic Reactions Using RBL-2H3 Cells

[Table 23]

Experiment 6: Mouse collagen-induced arthritis model (preventive effect)

Experiment 7: Mouse collagen-induced arthritis model (therapeutic effect)

Experimental Example 9: Mouse Antigen-Induced Dermatitis Model

[Table 24]

Experimental Example 10: Guinea Pig Antigen-Induced Rhinitis Model

Group 1: Comparative compound 1 (280 mg/kg) was administered orally once daily.

[Table 25]

Experimental Example 12: Mouse model of systemic lupus erythematosus

Claims

1. The use of a compound of general formula (I) or a salt thereof in the manufacture of a preventive and/or therapeutic agent for an immune disease associated with BTK, wherein the immune disease is an allergic disease, and the allergic disease is urticaria.

In the formula, X is acrylonitrile, pyrrolidinyl or piperidinyl;

Y represents -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

W and Z independently represent N or CH, respectively;

n is 0;

_R1 represents an amino group;

One of _R2 and _R3 is a hydrogen atom or a C1-C6 alkyl group, and the other is a hydrogen atom, a halogen atom, a C1-C6 alkyl group, a halo-C1-C6 alkyl group, a C1-C4 alkoxy-substituted C1-C6 alkyl group, a C1-C6 alkoxy group, a phenyl group, a 4- to 6-membered monocyclic unsaturated heterocyclic group containing one sulfur atom, or a cyano group.

When Y is -C( _R4 ) = C( _R5 )( _R6 ),

_R4 is a hydrogen atom or a C1-C6 alkyl group. Either _R5 or _R6 must be a hydrogen atom, and the other is a hydrogen atom, a C1-C6 alkyl group, or a C1-C6 alkyl group substituted with two C1-C6 alkyl groups and an amino group. In the C1-C6 alkyl group substituted with two C1-C6 alkyl groups and an amino group substituted with two C1-C6 alkyl groups, the C1-C6 alkyl group can form a 4-8 membered heterocyclic alkyl group together with the nitrogen atom to which it is attached.

When Y is -C≡C-R ₇ ,

_R7 is a hydrogen atom or a C1-C6 alkyl group.

2. The use as described in claim 1, characterized in that:

In general formula (I),

X is 1,3-acrylidine, 1,3-pyridinealkyl, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a C1-C4 alkyl group, and the other is a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a halo-C1-C4 alkyl group, a C1-C4 alkoxy-substituted C1-C4 alkyl group, a C1-C4 alkoxy group, a phenyl group, a 4- to 6-membered monocyclic unsaturated heterocyclic group containing one sulfur atom, or a cyano group.

When Y is -C( _R4 ) = C( _R5 )( _R6 ),

When Y is -C≡C-R ₇ ,

R ₇ is a hydrogen atom or a C1-C4 alkyl group.

3. The use as described in claim 1, characterized in that:

In general formula (I),

X is 1,3-acrylidine, 1,3-pyridinealkyl, or 1,3-piperidine;

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C- _R7 ;

When Z is N, W is N; when Z is CH, W is either N or CH.

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a methyl group, and the other is a hydrogen atom, a halogen atom, a methyl group, a trifluoromethyl group, a methoxyethyl group, a methoxy group, a phenyl group, a 4-chlorophenyl group, a 2-thienyl group, or a cyano group.

When Y is -C( _R4 ) = C( _R5 )( _R6 ),

_R4 is a hydrogen atom or a methyl group, either _R5 or _R6 must be a hydrogen atom, and the other is a hydrogen atom, a methyl group, a dimethylaminomethyl group, a methylethylaminomethyl group, a diethylaminomethyl group, a methylisopropylaminomethyl group, a 1-piperidinylmethyl group, or a 1-pyrrolidinylmethyl group.

When Y is -C≡C-R ₇ ,

_R7 is a methyl group.

4. The use as described in claim 1, characterized in that:

In general formula (I),

(1) When Z is N and W is N.

X is 1,3-piperidineyl.

Y represents vinyl.

(2) When Z is CH and W is N.

X is 1,3-pyridinealkyl or 1,3-piperidineyl.

Y is -C( _R4 ) = C( _R5 )( _R6 ) or -C≡C-( _R7 ).

When Y is -C( _R4 ) = C( _R5 )( _R6 ),

_R4 is a hydrogen atom or a methyl group; either _R5 or _R6 must be a hydrogen atom, and the other must be a hydrogen atom, a methyl group, a dimethylaminomethyl group, a methylethylaminomethyl group, a diethylaminomethyl group, a methylisopropylaminomethyl group, a 1-piperidinylmethyl group, or a 1-pyrrolidinylmethyl group.

When Y is -C≡C-(R ₇ ),

_R7 is a methyl group.

(3) When Z is CH and W is CH

X is 1,3-adiazine-1,3-pyrrolidine-1,3-diphenylene ...

Y is -C(( _R4 ) = C( _R5 )( _R6 ),

_R4 is a hydrogen atom, either _R5 or _R6 must be a hydrogen atom, and the other is a hydrogen atom, dimethylaminomethyl, methylethylaminomethyl, diethylaminomethyl, methylisopropylaminomethyl, 1-piperidinylmethyl or 1-pyrrolidinylmethyl.

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom or a methyl group, and the other is a hydrogen atom, a halogen atom, a trifluoromethyl group, a methoxyethyl group, a phenyl group, a 2-thiophene group, or a cyano group.

5. The use as described in claim 1, characterized in that:

In general formula (I),

X is 1,3-piperidinyl;

Y represents vinyl;

Z stands for CH;

W is N;

n is 0;

_R1 is an amino group;

One of _R2 and _R3 is a hydrogen atom, and the other is a hydrogen atom, a halogen atom, or a cyano group.

6. The use as described in claim 1, characterized in that:

The compounds represented by general formula (I) are selected from the following compounds:

(1)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(2)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(5-bromobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(3)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(4)(R)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－1－(1－methacryloylpiperidin-3－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide,

(5)(R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(6)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-cyanobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(7)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-methoxybenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(8)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N－(5-(2-methoxyethyl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(9)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(Oxazo[4,5-b]pyridin-2-yl)－1H-pyrazo[3,4-d]pyrimidine-3-carboxamide),

(10)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(4-methylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(11)(R)－4－amino－N－(5－fluorobenzo[d]oxazol-2－yl)－1－(1－methacryloylpiperidin-3－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide,

(12)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(5-fluorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(13)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(14) (R,E)-4-amino-N-(benzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(15) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(16) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(ethyl(methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(17) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(diethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(18) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(19) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyrrolidone-1-yl)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(20)(R,E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－1－(1－(4－(piperidin-1－yl)but-2－enoyl)piperidin-3－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide,

(21) (R,E)-4-amino-N-(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(22)(R)－4－amino－N－(benzo[d]oxazol-2－yl)－1－(1－(but-2-ynyl)piperidin-3－yl)－1H－pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(23)(R)－1－(1-Acryloylpiperidin-3-yl)－4-amino-N-(5,6-dimethylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(24)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(25) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(26) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(3-methylbut-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(27)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N-(benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(28)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N－(5-(thiophen-2-yl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(29)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N-(5-methylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(30)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N－(5-fluorobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(31)(R)－1－(1－acryloylpyrrolidone-3－yl)－4－amino-N－(5－(4－chlorophenyl)benzo[d]oxazol-2－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide,

(32) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(33)(R,E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－1－(1－(4－(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3－yl)－1H－pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(34) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(diethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(35)(R,E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－1－(1－(4－(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3－yl)－1H－pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(36) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(pyrrolidine-1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(37) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1-(1-(4-(piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(38)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N－(5-methoxybenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(39)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N－(5-cyanobenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(40)(R)－1－(1－acryloylpyrrolidone-3－yl)－4－amino-N－(5－(2－methoxyethyl)benzo[d]oxazol-2－yl)－1H－pyrazolo[3,4－d]pyrimidine-3－carboxamide,

(41)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(42) (R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(43)(R)－1－(1-Acryloylpyrrolidone-3-yl)－4-amino-N－(5-(trifluoromethyl)benzo[d]oxazol-2-yl)－1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(44) (R,E)-4-amino-N-(5-(trifluoromethyl)benzo[d]oxazol-2-yl)-1-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(45) 1-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidine-3-carboxamide,

(46) 7-(1-Acryloylacrylidine-3-yl)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(47)(E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－7－(1－(4－(dimethylamino)but-2-enoyl)acetidin-3－yl)7H－pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(48)(R)－7－(1－acryloylpyrrolidine-3－yl)－4－amino-N－(5－chlorobenzo[d]oxazol-2－yl)－7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide,

(49) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(50)(R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(51)(R,E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－7－(1－(4－(diethylamino)but-2-enoyl)pyrrolidine-3－yl)7H－pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(52) (R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(53)(R,E)－4－amino－N－(5－chlorobenzo[d]oxazol-2－yl)－7－(1－(4－(pyrrolidine-1－yl)but-2－enoyl)pyrrolidine-3－yl)7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide,

(54)(R,E)-4-amino-N-(5-chlorobenzo[d]oxazol-2-yl)-7-(1-(4-(piperidin-1-yl)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(55)(R)－7－(1－acryloylpyrrolidine-3－yl)－4－amino-N－(5－phenylbenzo[d]oxazol-2－yl)－7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide,

(56) (R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(dimethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(57)(R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(ethyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(58) (R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(diethylamino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(59) (R,E)-4-amino-N-(5-phenylbenzo[d]oxazol-2-yl)-7-(1-(4-(isopropyl(methyl)amino)but-2-enoyl)pyrrolidine-3-yl)7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide,

(60)(R,E)－4－amino－N－(5－phenylbenzo[d]oxazol-2－yl)－7－(1－(4－(pyrrolidine-1－yl)but-2－enoyl)pyrrolidine-3－yl)7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide,

(61)(R,E)－4－amino－N－(5－phenylbenzo[d]oxazol-2－yl)－7－(1－(4－(piperidin-1－yl)but-2－enoyl)pyrrolidine-3－yl)7H－pyrrolo[2,3－d]pyrimidine-5－carboxamide.