HK1118016B

HK1118016B - Immunosuppressive agent and anti-tumor agent comprising heterocyclic compound as active ingredient

Info

Publication number: HK1118016B
Application number: HK08109143.9A
Authority: HK
Inventors: 和彦春田; 信一矢口; 俊行松野; 良雄土田; 哲夫渡边; 君友吉冈; 龙五油井
Original assignee: 全药工业株式会社
Priority date: 2005-03-11
Filing date: 2006-03-13
Publication date: 2013-03-22

Description

Immunosuppressant and antitumor agent containing heterocyclic compound as active ingredient

Technical Field

The present invention relates to a novel immunosuppressant, and more particularly to an immunosuppressant containing a heterocyclic compound of a specific structure as an active ingredient. The invention also relates to novel compounds of the above heterocyclic compounds and to the use of these novel compounds as antitumor agents.

Background

In general, conditions in which immunosuppressants may be used include a number of autoimmune diseases, for example rejection after organ or tissue transplantation, graft-versus-host disease after bone marrow transplantation, inflammatory bowel diseases such as ulcerative colitis or crohn's disease, inflammatory or allergic skin diseases such as psoriasis or atopic dermatitis, inflammatory or allergic respiratory disorders such as chronic obstructive pulmonary disease or asthma, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, sjogren's syndrome and the like. In addition, immunosuppressive agents such as cyclophosphamide or methotrexate are also used in the treatment of hematological neoplasms, such as multiple myeloma, malignant lymphoma, leukemia, and the like. Furthermore, in the case of treating a disorder having an enhanced immune function characteristic associated with an infection such as sepsis, an immunosuppressant may also be used in combination with an antibiotic (non-patent document 1).

Therefore, in clinical practice, many immunosuppressive agents are currently used as therapeutic agents for the above-mentioned conditions. However, according to the present circumstances, there are many problems to be improved because a sufficient therapeutic effect cannot be obtained and unexpected side effects may occur.

Various cells and factors such as T lymphocytes, B lymphocytes are known to be involved in the induction of immune responses. Since cyclosporine and tacrolimus (tacrolimus) currently used in organ transplantation and the like have limited effects on T cells, there is a need for immunosuppressive agents that act on a wider range of immune mechanisms, have fewer side effects in clinical applications, and simultaneously act on various cells involved in the disorder.

Here, "a plurality of cells involved in the disorder" is not limited to immune cells, i.e., T cells, B cells, monocytes, macrophages, NK cells, NKT cells, dendritic cells, neutrophils, basophils, eosinophils, mast cells, and the like. They should include cells whose function is affected by humoral factors released from immune cells or membrane receptors on immune cells. Examples of such cells include, but are not limited to, platelets, vascular endothelial cells, synovial cells, osteoclasts, osteoblasts, chondrocytes, tracheal epithelial cells, and the like. In addition, in the case where the humoral factor is an autoantibody, a cell expressing a target antigen is also included.

Regarding the benzimidazole ring-substituted s-triazine [1, 3, 5-triazine ] derivatives and pyrimidine derivatives, the present inventors studied their cytostatic activity against solid tumors, carried out the synthesis of a large number of such compounds, and confirmed the relationship between antitumor activity and chemical structure (see patent documents 1, 2, 3, 4 and 5).

In particular, it was found that s-triazine derivatives and pyrimidine derivatives having a specific substituent at the 2-position of the benzimidazole ring exhibit enhanced cytostatic activity against solid tumors (see patent documents 3, 4 and 5). Methods for producing these derivatives are described in these patent documents, but not limited to these methods, and various reactions such as alkylation, alkylcarbonylation and the like may be introduced in the final product to make the product as a final compound.

Non-patent document 1: t.munster et.al.clin.exp.rheumato, 17 (suppl.18): S29-S36 (1999);

patent document 1: WO 99/05138 pamphlet

Patent document 2: WO 00/43385 pamphlet

Patent document 3: WO 02/088112 pamphlet

Patent document 4: WO 2004/037812 pamphlet

Patent document 5: WO 2005/095389 pamphlet

Problems to be solved by the invention

The present inventors have demonstrated that the above-mentioned compounds particularly inhibit phosphatidylinositol 3-kinase (PI3K) activity (non-patent document 2). PI3K is an enzyme that phosphorylates Phosphatidylinositol (PI) on cell membranes and is classified into three subfamilies according to its structure and substrate specificity. Within these subfamilies, the compounds of the invention specifically inhibit class I PI 3K. Class I PI3K phosphorylates PI, phosphatidylinositol 4-phosphate and phosphatidylinositol 4, 5-bisphosphate to produce phosphatidylinositol 3-phosphate, phosphatidylinositol 3, 4-bisphosphate and phosphatidylinositol 3, 4, 5-triphosphates, respectively. The phosphatidylinositol 3, 4, 5-triphosphate thus produced serves as an intracellular second messenger. Class I PI3K is expressed in a variety of cells and exhibits a broad spectrum of functions, such as cell proliferation, cell survival, glucose transport, cytoskeletal regulation, and the like. In PI3K knockout animals, development and signal transduction of cells such as B cells and T cells are hindered. In addition, degranulation of mast cells and migration of leukocytes are also hindered (non-patent document 3).

It is known that B cell proliferation caused by Lipopolysaccharide (LPS) or an anti-IgM antibody is inhibited by wortmannin or inhibitors of LY294002 and PI3K (non-patent document 4). Furthermore, wortmannin inhibits T cell proliferation induced by anti-CD 3 antibody and anti-CD 28 antibody (non-patent document 5).

Hematological neoplasms are characterized by spontaneous enhancement of cell division and inhibition of immune cell apoptosis. However, abnormalities in the PI3K cascade, such as a decrease in PTEN protein dephosphorylation of phosphatidylinositol 3, 4, 5-triphosphate and an increase in Akt phosphorylation, have been reported (non-patent document 6). Furthermore, it has been demonstrated that inhibition of PI3K can lead to inhibition of cell division and induction of apoptosis in various blood tumors (e.g., non-patent document 7).

Rheumatoid arthritis is a condition characterized by immune abnormalities and hypertrophy of synovial tissue. It is known that synovial tissue hypertrophy is caused by proliferation of synovial cells and inhibition of apoptosis thereof. In inflamed synovial tissue of patients with rheumatoid arthritis, the level of phosphorylated Akt is increased due to activation of PI3K (non-patent document 8). In addition, in vitro studies have shown that the inhibition of PI3K normalizes the proliferation of synovial cells and the inhibition of their apoptosis (non-patent document 9).

However, wortmannin and LY294002 were not put to clinical use due to their toxicity. Furthermore, despite the development of a large number of candidates with therapeutic potential for a wide range of conditions such as inflammation, cancer, etc. to exploit the inhibitory properties of PI3K, none have been put into clinical use. Therefore, there is a great need for immunosuppressive agents that normalize PI3K hyperactivity in a variety of cells involved in immune system disorders without exhibiting any toxicity to living individuals.

Non-patent document 2: S.Yaguchi et al, 96th Annual Meeting of the AACR, Anaheim, CA, USA. April 16-20, 2005, #1691

Non-patent document 3: R.Wetzker and C.Rommel, Current Pharmaceutical Design, 2004, 10, 1915-1922

Non-patent document 4: donahue and d.a.fruman, j.immunol.2003, 170, 5851-

Non-patent document 5: S.G.Ward et al, Eur J Immunol.1995, 25, 526-

Non-patent document 6: P.Workmann, biochem.Soc.Trans.2004, 32, 393-396

Non-patent document 7: s.uddin et al, biochem. biophysis. res. commun.2004, 320, 932-

Non-patent document 8: H.Zhang et al, Arthritis Rheum 2001, 44, 1555-

Non-patent document 9: miyashita et al, Biochem Biophys Res Commun2003, 312, 397-

Means for solving the problems

In view of the foregoing problems, the present inventors have conducted extensive studies on the heterocyclic compounds disclosed in patent documents 1, 2, 3, 4 and 5, predicting that some of them may be useful for a condition using an immunosuppressant, such as autoimmune diseases, organ transplantation, allergic diseases, hematological tumors, sepsis and the like. As a result, the present inventors have found that a heterocyclic compound represented by the following general formula (I) is effective, and have completed the present invention.

Accordingly, one aspect of the present invention provides an immunosuppressant comprising a heterocyclic compound represented by the general formula (I):

general formula (I)

Wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₁and R₂Two or any one of them represents a hydrogen atom, a hydroxyl group, a halogen, an amino group, C₁-C₆Alkylamino radical, C₁-C₆Alkoxy radical, C₁-C₆An alkyl group or a cyano group;

R₃represents a hydrogen atom, a difluoromethyl group, an amino groupRadical, C₁-C₆An alkylamino group, a methyl or hydroxymethyl group;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

R₆represents morpholino (by one or two C's)₁-C₆Alkyl group optionally substituted), pyrrolidinyl (by hydroxy C)₁-C₆Alkyl optionally substituted), piperidino (by one or two oxygen atoms, hydroxy groups, formyl or C₁-C₆Alkyl optionally substituted), piperazinyl (optionally substituted by one or two oxygen atoms, nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆The substituents of oxoalkyl, aromatic carbonyl, benzylcarbonyl and substituted carbamoyl radicals being optionally substituted) or 1, 4-diazepinyl (optionally substituted by one or two oxygen atoms, the nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆The substituents of oxoalkyl, aromatic carbonyl, benzylcarbonyl, and substituted carbamoyl groups are optionally substituted).

In this context, one embodiment provides that R in the formula (I)₁Or R₂Any of which is an immunosuppressive of hydroxyl groups. Another embodiment provides R in the formula (I)₁Or R₂Any of which is a hydroxyl group, and R₃Is an immunosuppressant for difluoromethyl. Another embodiment provides R in the formula (I)₁And R₂Are all hydrogen, and R₃Is an immunosuppressant for difluoromethyl. Another embodiment provides R in the formula (I)₆An immunosuppressant which is 4-acetylpiperazine.

As mentioned previously, the condition to be treated may be rejection and graft-versus-host disease, inflammatory bowel disease such as ulcerative colitis or crohn's disease, inflammatory or allergic skin disease such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as obstructive pulmonary disease or asthma; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; hematological tumors such as malignant lymphoma, multiple myeloma, chronic leukemia, acute leukemia, or myeloid leukemia; sepsis, fulminant hepatitis, etc.

In this context, one embodiment provides that R in the formula (I)₁Or R₂PI3K inhibitor in which either is a hydroxyl group. Another embodiment provides R in the formula (I)₁Or R₂Any of which is a hydroxyl group, and R₃PI3K inhibitor that is difluoromethyl. Another embodiment provides R in the formula (I)₁And R₂Are all hydrogen, and R₃PI3K inhibitor that is difluoromethyl. In addition, R in the general formula (I)₆An immunosuppressant which is 4-acetylpiperazine.

As mentioned previously, the condition to be treated may be rejection and graft versus host disease, inflammatory bowel disease such as ulcerative colitis or crohn's disease, inflammatory or allergic skin disease such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as obstructive lung disease or asthma; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; hematological tumors such as malignant lymphoma, multiple myeloma, chronic leukemia, or acute leukemia; sepsis, fulminant hepatitis, and the like.

According to the present invention, among the heterocyclic compounds of the general formula (I) used in the immunosuppressants, the structures of some compounds are novel. Thus in another aspect, the present invention provides a heterocyclic compound represented by the general formula (II):

wherein the content of the first and second substances,

n represents 0 to 2;

x represents a nitrogen atom or CH;

y represents- (CH)₂)n₁-, wherein n1 is 1 to 2;

R₃represents a hydrogen atom, a difluoromethyl group, an amino group, C₁-C₆An alkylamino group, a methyl or hydroxymethyl group;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

R₇represents a hydrogen atom, C₁-C₆Alkyl radical, formyl radical, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆Oxoalkyl, aromatic carbonyl, benzylcarbonyl or substituted carbamoyl.

For example, the compound according to formula (II) may be:

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-hydroxymethylbenzoimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (2, 2-dimethylmorpholino) pyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylpiperazin-1-yl) -2- (2-hydroxymethylbenzoimidazol-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) -piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) -piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) -piperazin-1-yl ] -6-morpholinopyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

4- [4- (2-furoyl) piperazin-1-yl ] -2- (2-hydroxymethylbenzoimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

4- [4- (2-furoyl) piperazin-1-yl ] -2- (2-hydroxymethylbenzoimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-hydroxymethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (2, 2-dimethylmorpholino) pyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -6-morpholinopyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-hydroxymethylbenzimidazol-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (cis-2, 3-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (trans-2, 3-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (cis-2, 3-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (trans-2, 3-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6- (2, 2-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6- (2, 2-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (cis-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (trans-2, 3-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (2, 2-dimethylmorpholino) -1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (cis-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (trans-2, 3-dimethylmorpholino) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6- (2, 2-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (5-oxo-1, 4-diazepin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (3-oxopiperazin-1-yl) -1, 3, 5-triazine; and

2- (2-difluoromethylbenzimidazol-1-yl) -6- (3, 5-dioxopiperazin-1-yl) -4-morpholino-1, 3, 5-triazine.

Furthermore, the present inventors have found a novel use of the novel compound represented by the above general formula (II) as an antitumor agent. Accordingly, another embodiment relates to an anticancer agent comprising the compound of the general formula (II) as an active ingredient. Disorders of interest may include, but are not limited to, lung cancer, prostate cancer, breast cancer, colon cancer, stomach cancer, pancreatic cancer, liver cancer, esophageal cancer, brain tumor, ovarian cancer, uterine cancer, malignant melanoma, kidney cancer, head and neck cancer, skin cancer, bladder cancer, osteogenic sarcoma, biliary tract cancer, vulval cancer, testicular tumor, penile cancer, colorectal cancer, mediastinal tumor, urothelial cancer, choriocarcinoma, soft tissue sarcoma, thyroid cancer, parathyroid cancer, adrenal cancer, malignant pheochromocytoma, blastoma, and the like.

Furthermore, the present invention relates to various embodiments as follows. The present invention relates to a method of immunosuppression comprising administering to a mammal a therapeutically effective amount of a heterocyclic compound represented by the general formula (I):

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₄and R₅Represents a hydrogen atomSeed or C₁-C₆An alkyl group;

Here, in one embodiment, R in the formula (I)₁Or R₂Any of which is a hydroxyl group. In another embodiment, R in formula (I)₁Or R₂Any of which is a hydroxyl group, and R₃Is difluoromethyl. In another embodiment, R in formula (I)₁And R₂Are all hydrogen, and R₃Is difluoromethyl. In another embodiment, R in formula (I)₆Is 4-acetyl piperazine.

Furthermore, the condition to be treated may be rejection and graft versus host disease, inflammatory bowel disease such as ulcerative colitis or crohn's disease, inflammatory or allergic skin disease such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as obstructive lung disease or asthma; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; hematological tumors such as malignant lymphoma, multiple myeloma, chronic leukemia, acute leukemia, and the like; sepsis, fulminant hepatitis, and the like.

Another embodiment relates to a method of treating a tumor comprising administering to a patient a therapeutically effective amount of a heterocyclic compound represented by the general formula (II):

wherein the content of the first and second substances,

n represents 0 to 2;

x represents a nitrogen atom or CH;

y represents- (CH)₂)n₁-, in which n₁Is 1 to 2;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In another embodiment, the present invention relates to an immunosuppressive composition comprising a therapeutically effective amount of a heterocyclic compound represented by the general formula (II):

wherein the content of the first and second substances,

n represents 0 to 2;

x represents a nitrogen atom or CH;

y represents- (CH)₂)n₁-, wherein n1 is 1 to 2;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In another embodiment, the present invention relates to a composition, in particular a pharmaceutical composition and more preferably an anti-tumor composition, comprising a therapeutically effective amount of a heterocyclic compound of formula (II) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Furthermore, the present invention relates to various embodiments as follows. The present invention relates to a PI3K inhibition method comprising administering to a mammal a therapeutically effective amount of a heterocyclic compound represented by the general formula (I):

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

Furthermore, the condition to be treated may be rejection and graft versus host disease; inflammatory bowel disease such as ulcerative colitis or crohn's disease; inflammatory or allergic skin diseases such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as obstructive lung disease or asthma; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; hematological tumors such as malignant lymphoma, multiple myeloma, chronic leukemia, acute leukemia, and the like; sepsis, fulminant hepatitis, and the like.

In another embodiment, the present invention relates to a PI3K inhibitory composition comprising a therapeutically effective amount of a heterocyclic compound represented by the general formula (I):

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In another embodiment, the present invention relates to a composition, in particular a pharmaceutical composition and more preferably a PI3K inhibitory composition, comprising a therapeutically effective amount of a heterocyclic compound of formula (II) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Furthermore, the present invention relates to the use of a heterocyclic compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of an immunological disease, or as an immunosuppressant:

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In addition, the condition to be treated may be rejection and graft versus host disease; inflammatory bowel disease such as ulcerative colitis or crohn's disease; inflammatory or allergic skin diseases such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as obstructive lung disease or asthma; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; hematological tumors such as malignant lymphoma, multiple myeloma, chronic leukemia, acute leukemia, and the like; sepsis, fulminant hepatitis, and the like.

In another embodiment, the present invention relates to the use of a heterocyclic compound represented by the general formula (II) or a pharmaceutically acceptable salt thereof for the preparation of a composition for treating tumors:

wherein the content of the first and second substances,

n represents 0 to 2;

x represents a nitrogen atom or CH;

y represents- (CH)₂)n₁-, wherein n1 is 1 to 2;

R₃represents a hydrogen atom, a difluoromethyl group, an amino group, C₁-C₆Alkylamino radical, methyl radicalOr a hydroxymethyl group;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In another embodiment, the invention relates to the use of a heterocyclic compound of formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for inhibiting PI3K or as a PI3K inhibitor:

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

R₆represents morpholino (by one or two C's)₁-C₆Alkyl group optionally substituted), pyrrolidinyl (by hydroxy C)₁-C₆Alkyl optionally substituted), piperidino (by one or two oxygen atoms)Hydroxy, formyl or C₁-C₆Alkyl optionally substituted), piperazinyl (optionally substituted by one or two oxygen atoms, nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆The substituents of oxoalkyl, aromatic carbonyl, benzylcarbonyl and substituted carbamoyl radicals being optionally substituted) or 1, 4-diazepinyl (optionally substituted by one or two oxygen atoms, the nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆The substituents of oxoalkyl, aromatic carbonyl, benzylcarbonyl, and substituted carbamoyl groups are optionally substituted).

Here, in one embodiment, R in the formula (I)₁Or R₂Either is a hydroxyl group. In another embodiment, R in formula (I)₁Or R₂Any one is a hydroxyl group, and R₃Is difluoromethyl. In another embodiment, R in formula (I)₁And R₂Are all hydrogen, and R₃Is difluoromethyl. In another embodiment, R in formula (I)₆Is 4-acetyl piperazine.

In other embodiments, the modes and features of the invention will be apparent to those skilled in the art from the following detailed description.

Effects of the invention

The agent of the present invention is effective for the treatment of a disease due to hyperactivity of PI3K, such as autoimmune diseases, organ transplantation, allergic or inflammatory diseases, hematological tumors, sepsis, or the like, as well as tumors.

Brief description of the drawings

FIG. 1 is a graph showing the effect of test substances on the upregulation of CD69 expression and CD40L expression on human peripheral blood mononuclear cells induced by anti-human CD3 antibodies and anti-human CD28 antibodies.

FIG. 2 is a graph showing a comparison between the test substance-treated group and the control group in terms of changes in the volume of the hind paw of rats after the onset of adjuvant-induced arthritis.

FIG. 3 is a graph showing the comparison between the test substance-treated group and the control group in terms of the change in the arthritis score after the onset of collagen-induced arthritis.

FIG. 4 is a graph showing the comparison between the test substance-treated group and the control group in the human B lymphoma xenograft model.

Best mode for carrying out the invention

The heterocyclic compound used in the present invention is a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₁and R₂Two or any one of them represents a hydrogen atom, a hydroxyl groupHalogen, amino group, C₁-C₆Alkylamino radical, C₁-C₆Alkoxy radical, C₁-C₆An alkyl group or a cyano group;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

In the above formula, "C₁-C₆"represents, without any limitation, a group having 1 to 6 carbon atoms. "C₁-C₆Alkyl "includes straight or branched chain alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, n-hexyl, and the like. "C₁-C₆Alkoxy "includes alkoxy groups having a straight or branched chain, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxyAnd so on. "hydroxy group C₁-C₆Alkyl "means having the same" C "as above₁-C₆Alkyl "a group in which any carbon atom in the group defined is bonded to a hydroxyl group.

When the above-mentioned heterocyclic compounds have an asymmetric carbon atom in the structure, isomers derived from the asymmetric carbon atom and mixtures thereof (racemic compounds) exist, and any of these compounds is included in the compounds of the present invention.

In addition, the heterocyclic compound used as the active ingredient of the present invention may be in the form of an acid addition salt as a pharmaceutically acceptable salt. Suitable acid addition salts include inorganic acid salts such as hydrochloride, sulfate, hydrobromide, nitrate, phosphate and the like and organic acid salts such as acetate, oxalate, propionate, glycolate, lactate, pyruvate, malonate, succinate, maleate, fumarate, malate, tartrate, citrate, benzoate, cinnamate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, salicylate and the like.

The compounds useful as the effective ingredient of the present invention include, but are not limited to, the following heterocyclic compounds:

2- (2-methylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 1)

2- (benzimidazol-1-yl) -4- (trans-2, 3-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine (Compound 2)

4, 6-dimorpholino-2- (2-hydroxymethylbenzoimidazol-1-yl) -1, 3, 5-triazine (Compound 3)

2- (2-Difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 4)

2- (2-aminobenzimidazol-1-yl) -4, 6-dimorpholinopyrimidine (Compound 5)

2- (2-Aminobenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 6)

2- (2-Difluoromethylbenzimidazol-1-yl) -4-piperidino-6-morpholino-1, 3, 5-triazine (Compound 7)

2- (6-amino-2-difluoromethylbenzimidazol-1-yl) -4- (cis-2, 3-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine (Compound 8)

2- (2-difluoromethyl-6-ethoxybenzoimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 9)

2- (2-difluoromethyl-4-methylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 10)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4-morpholino-6- (2, 6-dimethylmorpholino) pyrimidine (Compound 11)

2- (2-difluoromethyl-5-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 12)

2- (6-amino-2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine (Compound 13)

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 14)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 15)

2- (2-difluoromethyl-6-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 16)

2- (5-amino-2-difluoromethylbenzimidazol-1-yl) -4, 6-bis (2, 6-dimethyl-morpholino) pyrimidine (Compound 17)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4, 6-dimorpholinopyrimidine (Compound 18)

2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethyl-morpholino) -6-morpholino-1, 3, 5-triazine (Compound 19)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethyl-pyrrolidin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 20)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholinopyrimidine (Compound 21)

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (3, 3-dimethylmorpholino) -6-morpholinopyrimidine (Compound 22)

2- (2-difluoromethyl-5-methoxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 23)

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (cis-2, 6-dimethyl-morpholino) -6-morpholinopyrimidine (Compound 24)

2- (2-Difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (5-oxo-1, 4-diazepin-1-yl) -1, 3, 5-triazine (Compound 25)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- (4-hydroxypiperidin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 26)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- [4- (2-hydroxyethyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine (Compound 27)

4- (4-Acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 28)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- (4-benzylcarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 29)

4- (4-Acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 30)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- [4- (2-furoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine (Compound 31)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine (Compound 32)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- (4-methoxyacetylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 33)

2- (2-Difluoromethylbenzimidazol-1-yl) -4- [4- (3-hydroxypropyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine (Compound 34)

2- (2-Difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (4-propionylpiperazin-1-yl) -1, 3, 5-triazine (Compound 35)

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholinopyrimidine (Compound 36)

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (cis-2, 3-dimethyl-morpholino) -6-morpholinopyrimidine (Compound 37)

2- (2-difluoromethyl-4-ethoxybenzoimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholinopyrimidine (Compound 38)

As demonstrated in the examples described below, the agent of the present invention inhibits activation of T cells and B cells induced by Con a, LPS, anti-IgM antibody, anti-CD 3 antibody + anti-CD 28 antibody, and thus exhibits PI3K inhibitory behavior on immune cells. Therefore, the medicament of the present invention can be used for the treatment and prevention of immune system disorders due to hyperactivity of PI 3K.

Immune system disorders due to hyperactivity of PI3K may be: autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; organ dysfunction associated with autoimmune diseases such as uveitis, glomerulonephritis, thyroiditis, pancreatitis, bone destruction, etc.; rejection after tissue transplantation, graft versus host disease after bone marrow transplantation; inflammatory bowel disease such as ulcerative colitis or crohn's disease; inflammatory or allergic skin diseases such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as chronic obstructive pulmonary disease or asthma; allergic conjunctivitis or rhinitis; hematological tumors derived from immune cells such as B cell lymphoma, T cell lymphoma, myeloid leukemia; sepsis triggered by gram negative bacterial or coronavirus infection, severe acute respiratory syndrome, fulminant hepatitis, and the like.

Although the agent of the present invention can be applied to mammals such as human, dog, cat, rabbit, hamster, rat, mouse, etc., the administration schedule, preparation and dosage applied to human will be specifically explained below.

The agent of the present invention can be administered orally or parenterally, and tablets, coated tablets, powdered drugs, granules, capsules, microcapsules, slurries and the like can be used as a dosage form for oral administration, while eye drops, inhalants, injectable forms (including lyophilized preparations for injection dissolved at the time of application), suppositories, poultices and the like can be used as a dosage form for parenteral administration. The formulation of these dosage forms may be affected using pharmaceutically acceptable excipients, such as lactose, sucrose, starch, dextrin, crystalline cellulose, kaolin, calcium carbonate, talc, magnesium stearate, distilled water or saline, binders, lubricants, disintegrants, suspending agents, emulsifiers, preservatives, stabilizers and dispersants.

When used in oral dosage forms, the dose of the effective ingredient varies depending on the symptoms, age, body weight, etc. of a patient, but for an adult having a body weight of 60kg, a daily dose of 10 to 500mg may be administered in the form of 2 to 3 parts. In addition, in the case of ophthalmic solutions, inhalation to the lungs or nasal cavity, and injection into inflamed joint cavities, the dosage also varies depending on the symptoms of the patient, but for adults, a daily dosage of 1-100 μ g may be administered in 2-3 parts.

Examples

Preparation examples

Some examples of the heterocyclic compounds represented by the general formula (I) are prepared according to the methods disclosed in examples of patent documents 3, 4 and 5, and are described below.

The synthesis was carried out with reference to patent documents 1 to 3.

Preparation of example 1

2- (2-methylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 1)

Melting point: 218-20 deg.C (decomposition)

NMR(CDCl₃)δ：3.03(3H，s)，3.7-3.9(16H，m)，7.2-7.4(2H，m)，7.7-7.8(1H，m)，8.1-8.3(1H，m)

MS m/z：381(M⁺)

Preparation of example 2

Melting point: 147 deg.C, 150 deg.C

NMR(CDCl₃)δ：1.1-1.5(6H，m)，2.7-3.0(1H，m)，3.4-3.6(1H，m)，3.7-4.0(8H，m)，4.1-4.3(1H，m)，4.4-4.7(2H，m)，7.3-7.4(2H，m)，7.7-7.9(1H，m)，8.3-8.4(1H，m)，8.98(1H，s)

Preparation of example 3

Melting point: 208 + 210 deg.C (decomposition)

NMR(CDCl₃)δ：3.7-3.9(16H，m)，4.59(1H，t，J＝6Hz)，5.15(2H，d，J＝7Hz)，7.2-7.4(2H，m)，7.7-7.8(1H，m)，8.3-8.4(1H，m)

MS m/z：397(M⁺)

Preparation of example 4

Melting point: 211 ℃ C. 214 ℃ C

NMR(CDCl₃)δ：3.79(8H，t，J＝4Hz)，3.88(8H，t，J＝4Hz)，7.3-7.4(2H，m)，7.56(1H，t，J＝53Hz)，7.88(1H，d，J＝7Hz)，8.32(1H，d，J＝7Hz)

MS m/z：417(M⁺)

Preparation of example 5

2- (2-aminobenzimidazol-1-yl) -4, 6-dimorpholinopyrimidine (Compound 5)

Melting point: 237 plus 239 DEG C

NMR(CDCl₃)δ： 3.59(8H，t，J＝5Hz)，3.84(8H，t，J＝5Hz)，5.46(1H，s)，6.65(2H，brs)，7.06(1H，t，J＝7Hz)，7.18(1H，t，J＝7Hz)，7.37(1H，d，J＝7Hz)，8.1(1H，d，7Hz)

MS m/z：381(M⁺)

Preparation of example 6

2- (2-Aminobenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine (Compound 6)

Melting point: 298 + 300 deg.C (decomposition)

NMR(CDCl₃)δ：3.7-3.9(16H，m)，6.74(2H，brs)，7.05(1H，t，J＝7Hz)，7.20(1H，t，J＝7Hz)，7.39(1H，d，J＝7Hz)，8.20(1H，d，7Hz)

MS m/z：382(M⁺)

Preparation of example 7

Melting point: 190 ℃ to 192 DEG C

NMR(CDCl₃)δ：1.5-1.8(6H，m)，3.7-3.9(12H，m)，7.3-7.5(2H，m)，7.61(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.34(1H，d，8Hz)

MS m/z：415(M⁺)

Preparation of example 8

Melting point: 220 + 222 deg.C (decomposition)

NMR(CDCl₃)δ：1.22(3H，d，J＝9Hz)，1.26(3H，d，J＝9Hz)，3.1-3.4(1H，m)，3.5-4.1(11H，m)，4.3-4.5(1H，m)，4.5-4.7(1H，m)，6.77(1H，dd，J＝2Hz，J＝9Hz)，7.49(1H，t，J＝54Hz)，7.62(1H，d，J＝9Hz)，7.64(1H，d，J＝2Hz)

MS m/z：460(M⁺)

Preparation of example 9

Melting point: 222 ℃ and 224 DEG C

NMR(CDCl₃)δ：1.46(3H，t，J＝7Hz)，3.7-3.9(16H，m)，4.08(2H，q，J＝7Hz)，7.00(1H，dd，J＝9Hz，3Hz)，7.52(1H，t，J＝54Hz)，7.74(1H，d，J＝9Hz)，7.89(1H，d，J＝3Hz)

MS m/z：461(M⁺)

Preparation of example 10

Melting point: 259 ℃ 260 DEG C

NMR(CDCl₃)δ：2.72(3H，s)，3.7-3.9(16H，m)，7.1-7.5(2H，m)，7.56(1H，t，J＝54Hz)，8.15(1H，d，8Hz)

MS m/z：431(M⁺)

Preparation of example 11

Melting point: 265 ℃ 267 DEG C

¹H NMR(CDCl₃)δ：3.7-3.9(16H，m)，3.86(3H，s)，7.02(1H，dd，J＝3，9Hz，)，7.52(1H，t，J＝53Hz，)，7.75(1H，d，J＝9Hz，)，7.91(1H，d，J＝3Hz，)

MS m/z：447(M⁺)

Preparation of example 12

Melting point: 272 sub-274 deg.C

NMR(CDCl₃)δ：3.7-3.9(16H，m)，7.26-7.29(2H，m)，7.54(1H，t，J＝54Hz)，8.20(1H，d，8Hz)

MS m/z：433(M⁺)

Example 13

Melting point: 226 ℃ in 227 deg.C (decomposition)

NMR(CDCl₃)δ：1.28(6H，s)，3.6-3.8(14H，m)，6.7-6.8(1H，m)，7.2-7.7(3H，m)

MS m/z：460(M⁺)

Example 14

Melting point: 214 ℃ 216 ℃ (decomposition)

NMR(CDCl₃)δ：3.7-3.9(16H，m)，4.48(2H，brs)，6.63(1H，d，J＝8Hz)，7.21(1H，t，J＝8Hz)，7.55(1H，t，J＝54Hz)，7.64(1H，d，J＝8Hz)

MS m/z：432(M⁺)

Preparation of example 15

Melting point: higher than 250 DEG C

NMR(DMSO-d₆)δ：3.70-3.90(16H，m)，6.76(1H，d，J＝8Hz)，7.73(1H，t，J＝8Hz)，7.70(1H，t，J＝54Hz)，7.74(1H，d，J＝8Hz)，10.24(1H，brs)

MS m/z：433

Preparation of example 16

Melting point: higher than 250 DEG C

NMR(DMSO-d₆)δ：3.70-3.90(16H，m)，6.86(1H，d，J＝8Hz)，7.61(1H，d，J＝8Hz)，7.70(1H，t，J＝54Hz)，7.73(1H，s)，9.81(1H，brs)

MS m/z：433

Preparation of example 17

2- (5-amino-2-difluoromethylbenzimidazol-1-yl) -4, 6-bis (2, 6-dimethylmorpholino) -pyrimidine (Compound 17)

Melting point: 157 ℃ C. & 160℃.

NMR(CDCl₃)δ：1.30(6H，d，J＝9Hz)，2.6-2.8(4H，m)，3.6-4.2(8H，m)，5.45(1H，s)，6.7-6.8(1H，m)，7.5-7.7(2H，m)，7.42(1H，t，J＝53Hz)

MS m/z：487(M⁺)

Preparation of example 18

Melting point: higher than 250 DEG C

NMR(DMSO-d₆)δ：3.60-3.80(16H，m)，5.98(1H，s)，6.72(1H，d，J＝8Hz)，7.22(1H，t，J＝8Hz)，7.62(1H，d，J＝8Hz)，7.65(1H，t，J＝54Hz)，10.17(1H，brs)

MS m/z：432

Preparation of example 19

2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine (Compound 19)

(1) 6-amino-4-chloro-2-difluoromethylbenzimidazole (500mg, 2.3mmol) was dissolved in acetone (50ml), and 2, 4-dichloro-6-morpholino-1, 3, 5-triazine (542mg, 2.3mmol) was added at-15 ℃ followed by further addition of potassium carbonate (500 mg). After gradually raising the temperature to room temperature, the mixture was stirred at room temperature for 5 hours. The solvent was evaporated in vacuo and the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate 1: 4) to give 2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4-chloro-6-morpholino-1, 3, 5-triazine (272mg, yield 28%).

(2) The resulting 2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4-chloro-6-morpholino-1, 3, 5-triazine (150mg, 0.36mmol) was dissolved in DMF (6ml), to which was added 2, 2-dimethylmorpholine hydrochloride (150mg, 1.0mmol) at-15 ℃ and then further added potassium carbonate (500 mg). After stirring at room temperature overnight, water was added to the reaction mixture, extracted several times with ethyl acetate, washed with brine, and dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate 1: 2) to give the title compound (130mg, yield 73%) as colorless crystals.

Melting point: 238 deg.C (decomposition)

NMR(CDCl₃)δ：1.27(6H，s)，3.68(2H，s)，3.7-3.9(12H，m)，6.82(1H，d，J＝2.3Hz)，7.42(1H，dt，J＝9.6Hz，J＝53Hz)，7.50(1H，d，J＝2.3Hz)

MS m/z：494(M⁺)

Preparation of example 20

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 20)

Melting point: 245 ℃ (decomposition)

NMR(CDCl₃)δ：1.9-2.1(4H，m)，3.5-4.0(12H，m)，4.7-4.8(1H，m)，5.1-5.3(1H，m)，6.89(1H，d，J＝9Hz)，7.30(1H，t，J＝9Hz)，7.50(1H，brs)，7.55(1H，t，J＝54Hz)，7.83(1H，d，J＝9Hz)

MS m/z：447(M⁺)

Preparation of example 21

Melting point: 185 ℃ C., 187 ℃ C

NMR(CDCl₃)δ：1.29(6H，s)，3.48(2H，s)，3.59-3.64(6H，m)，3.81-3.87(6H，m)，5.47(1H，s)，6.86(1H，m)，7.26-7.32(1H，m)，7.49(1H，t，J＝53Hz)，7.72(1H，d，8Hz)

MS m/z：460(M⁺)

Preparation of example 22

Melting point: 204 ℃ and 206 DEG C

NMR(CDCl₃)δ：1.48(6H，s)，3.50(2H，s)，3.6-3.8(6H，m)，3.8-4.0(6H，m)，5.76(1H，s)，6.68(1H，d，J＝7Hz)，7.29(1H，d，J＝7Hz)，7.49(1H，t，J＝54Hz)，7.66(1H，d，7Hz)

MS m/z：460(M⁺)

Preparation of example 23

Melting point: 206- & ltD & gt 207 & deg.C

NMR((CD₃)₂CO)δ：1.17(6H，d，J＝6Hz)，2.5-2.8(4H，m)，3.6-4.4(10H，m)，5.95(1H，s)，6.77(1H，d，J＝8Hz)，7.23(1H，t，J＝8Hz)，7.66(1H，t，J＝53Hz)，7.75(1H，d，J＝8Hz)，8.9(1H，s)

MS m/z：447(M⁺)

Preparation of example 24

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (cis-2, 6-dimethylmorpholino) -6-morpholinopyrimidine (Compound 24)

(1) 2-difluoromethyl-4-methoxybenzimidazole (9.03g, 45.6mmol) was dissolved in DMF (100ml), 60% NaH (1.82g, 45.6mmol) was added and stirred for 30 min. The reaction mixture was added to a solution of 2, 4, 6-trichloropyrimidine (15.7g, 92.1mmol) in DMF (100ml) under ice-cooling, stirred on an ice bath for 30 minutes, and then further stirred at room temperature for 2 hours. Water was added to the reaction mixture, and the precipitated crystals were filtered, washed thoroughly with hexane and ether, and then air-dried to give 2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4, 6-dichloropyrimidine (12.3g, yield 78%).

(2) The obtained 2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4, 6-dichloropyrimidine (12.3g, 35.7mmol) was dissolved in DMF (150ml), and cis-2, 6-dimethylmorpholine (6.63ml, 53.7mmol) was added at room temperature, followed by further addition of potassium carbonate (7.35 g). After stirring at room temperature for 30 minutes, water was added to the reaction mixture, extracted several times with ethyl acetate, washed with brine, and dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was washed thoroughly with hexane then ether in order, then air dried to give 4-chloro-2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -6- (cis-2, 6-dimethyl-morpholino) pyrimidine (14.4g, yield 95%).

(3) To the obtained 4-chloro-2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -6- (cis-2, 6-dimethylmorpholino) pyrimidine (14.4g, 34mmol), morpholine (275ml, 3.15mol) was added, and stirred at room temperature for 30 minutes, followed by further stirring at 80 ℃ for 30 minutes. Water was added to the reaction solution, and the precipitated crystals were filtered, followed by thorough washing with hexane, ether and ethyl acetate in this order, and air-drying to give 2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (cis-2, 6-dimethyl-morpholino) -6-morpholinopyrimidine (13.7g, yield 86%).

Melting point: 180 ℃ and 181 DEG C

NMR(CDCl₃)δ：1.28(6H，d，J＝6Hz)，2.6-2.7(2H，m)，3.6-3.7(6H，m)，3.80-3.86(4H，m)，4.04(3H，s)，4.10-4.14(2H，m)，5.49(1H，s)，6.78(1H，d，J＝8Hz)，7.32(1H，d，J＝8Hz)，7.41(1H，t，J＝52Hz)，7.77(1H，d，J＝8Hz)

MS m/z：474(M⁺)

Preparation of example 25

Melting point: 235-37 deg.C

¹H NMR(CDCl₃)δ：2.7-2.8(2H，m)，3.4-3.5(2H，m)，3.8-4.2(12H，m)，5.97(1H，brs)，7.2-7.5(2H，m)，7.52(1H，t，J＝54Hz)，7.8-8.0(1H，m)，8.2-8.4(1H，m)

MS m/z：444(M⁺)

Preparation of example 26

Melting point: 219-21 deg.C

¹H NMR(CDCl₃)δ：3.4-3.5(2H，m)，3.7-4.1(16H，m)，7.3-7.5(2H，m)，7.59(1H，t，J＝50Hz)，7.8-8.0(1H，m)8.3-8.4(1H，m)

MS m/z：431(M⁺)

Preparation of example 27

Melting point: 174-77 deg.C

¹H NMR(CDCl₃)δ：2.6-2.7(8H，m)，3.6-3.9(12H，m)，3.91(1H，br)，7.3-7.5(2H，m)，7.58(1H，t，J＝54Hz)，7.9-8.0(1H，m)，8.3-8.4(1H，m)

MS m/z：460(M⁺)

See patent document 4.

Preparation of example 28

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholinopyrimidine

(1) 4-t-Butyldimethylsilyloxy-2-difluoromethylbenzimidazole (1.49g, 5.0mmol) was dissolved in DMF (10ml), 2, 4, 6-trichloropyrimidine (0.91g, 5.0mmol) was added at room temperature, followed by further addition of potassium carbonate (0.55g), and the mixture was stirred for 5 hours. Water was added to the reaction mixture, extracted several times with ethyl acetate, washed with brine, and then dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate 8: 1) to give 2- (4-tert-butyldimethylsilyloxy-2-difluoromethylbenzimidazol-1-yl) -4, 6-dichloropyrimidine (1.12g, yield 50%).

(2) The obtained 2- (4-tert-butyldimethylsilyloxy-2-difluoromethylbenzimidazol-1-yl) -4, 6-dichloropyrimidine (386mg, 0.87mmol) was dissolved in DMF (6ml), and 2-pyrrolidinemethanol (0.13ml, 1.3mmol) was added at room temperature, followed by further addition of potassium carbonate (179 mg). After the mixture was stirred at room temperature for 30 minutes, water was added to the reaction mixture, extracted several times with ethyl acetate, washed with brine, and dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate 1: 1) to give 2- (4-tert-butyldimethylsilyloxy-2-difluoromethylbenzimidazol-1-yl) -4- (2-hydroxy-methylpyrrolidin-1-yl) -6-chloropyrimidine (291mg, yield 64%).

(3) To the obtained 2- (4-tert-butyldimethylsilyloxy-2-difluoromethylbenzimidazol-1-yl) -4- (2-hydroxymethyl-pyrrolidin-1-yl) -6-chloropyrimidine (281mg, 0.54mmol) was added morpholine (4.4g, 50mmol), and the mixture was stirred at room temperature for 9 hours. To the reaction mixture was added water, extracted several times with ethyl acetate, washed with brine, and then dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate 2: 3) to give 2- (4-tert-butyldimethylsilyloxy-2-difluoromethylbenzimidazol-1-yl) -4- (2-hydroxy-methylpyrrolidin-1-yl) -6-morpholinopyrimidine (216mg, yield 72%).

2- (4-tert-Butyldimethylsilyloxy-2-difluoromethyl-benzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholinopyrimidine (213mg, 0.38mmol) was dissolved in anhydrous THF (7ml), tetra-n-butylammonium fluoride (0.4ml) (1M THF solution) was added at room temperature, and the mixture was stirred for 30 minutes. To the reaction mixture was added water, extracted several times with ethyl acetate, washed with brine, and dried over anhydrous magnesium sulfate. After evaporation of the solvent in vacuo, the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate: 1: 4) to give the title compound (101mg, yield 60%) as colorless crystals.

Melting point: 195 ℃ C. & 198 ℃ C

NMR(CDCl₃)δ：2.0-2.1(4H，m)，3.4-4.0(12H，m)，4.0-4.1(1H，m)，4.3-4.4(1H，m)，5.36(1H，s)，6.85(1H，d，J＝8Hz)，7.28(1H，t，J＝8Hz)，7.58(1H，brs)，7.58(1H，t，J＝54Hz)，7.73(1H，d，J＝8Hz)

MS m/z：446(M⁺)

Reference 4 realizes the preparation.

Preparation of example 29

2- (5, 6-dimethyl-2-difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine

Melting point: 217 ℃ 220-

¹H NMR(CDCl₃)δ：2.39(3H，s)，2.40(3H，s)，3.7-3.9(16H，m)，7.53(1H，t，J＝54Hz)，7.62(1H，s)，8.12(1H，s)

MS m/z：445(M⁺)

Preparation of example 30

2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholino-1, 3, 5-triazine (STK922)

Melting point: 256 deg.C (decomposition)

NMR (CD₃OD-CDCl₃(1∶1))δ：1.9-2.2(4H，m)，3.68(2H，s)，3.5-4.0(11H，m)，4.39(1H，brs)，6.84(1H，d，J＝2.1Hz)，7.58(1H，t，J＝53Hz)，7.64(1H，d，J＝2.1Hz)

MS m/z：480(M⁺)

Preparation of example 31

2- (4-chloro-2-difluoromethyl-5-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazi ne

Melting point: higher than 250 DEG C

NMR(CDCl₃)δ：3.7-3.9(16H，m)，5.63(1H，s)，7.15(1H，d，J＝9Hz)，7.51(1H，t，J＝53Hz)，8.14(1H，d，J＝9Hz)

MS m/z：467(M⁺)

Preparation of example 32

Synthesis of 4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 28)

A mixture of 6-chloro-2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-1, 3, 5-triazine (3.66g, 10mmol), 1-acetylpiperazine (1.40g, 11mmol), potassium carbonate (1.38g, 10mmol) and DMF (30ml) was stirred at room temperature for 16 hours. The reaction mixture was poured into water and extracted with dichloromethane. The extract was dried over anhydrous sodium sulfate, concentrated in vacuo, and the residue was purified by silica gel column to give 4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine (4.08g, 9.0mmol) as colorless crystals in 90% yield.

Naturally, triazine or pyrimidine derivatives bearing such piperazine groups can also be synthesized according to the following scheme.

For example, a mixture of 6-chloro-2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-1, 3, 5-triazine (3.66g, 10mmol), piperazine (3.45g, 40mmol) and acetone (50ml) was stirred at room temperature for 16 hours. The reaction mixture was poured into water and the precipitated crystals were filtered and washed with methanol to give 2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (piperazin-1-yl) -1, 3, 5-triazine (3.87g, 9.3mmol) as colorless crystals in 93% yield.

¹H NMR(CDCl₃)δ：3.8-4.1(16H，m)，7.3-7.5(2H，m)，7.59(1H，t，J＝54Hz)，7.9-8.0(1H，m)，8.3-8.4(1H，m)

MS m/z：416(M⁺)

To a mixture of 2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (piperazin-1-yl) -1, 3, 5-triazine (417mg, 1.0mmol) and THF (10ml) was added acetyl chloride (0.14ml, 2.0mmol) dropwise. The reaction mixture was stirred at room temperature for 22 hours. The reaction mixture was poured into water and extracted with dichloromethane. The extract was dried over anhydrous sodium sulfate, concentrated in vacuo, and the residue was purified by silica gel column to give 4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine (354mg, 7.7mmol) as a colorless crystalline product in 77% yield.

Melting point: 223 deg.C

¹H NMR(CDCl₃)δ：2.18(3H，s)，3.6-4.0(16H，m)，7.3-7.5(2H，m)，7.55(1H，t，J＝53.5Hz)，7.9-8.0(1H，m)，8.3-8.4(1H，m)

MS m/z：458(M⁺)

The following compounds were prepared in a similar manner to that for the preparation of example 34.

Preparation of example 33

2- (2-difluoromethylbenzimidazol-1-yl) -6- (4-formylpiperazin-1-yl) -4-morpholino-1, 3, 5-triazine

Melting point: 228-30 deg.C

¹H NMR(CDCl₃)δ：3.8-4.1(16H，m)，7.2-7.5(2H，m)，7.54(1H，t，J＝54Hz)，7.8-8.0(1H，m)，8.17(1H，s)，8.3-8.4(1H，m)

MS m/z：444(M⁺)

Preparation of example 34

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (3-oxopiperazin-1-yl) -1, 3, 5-triazine

Melting point: 255-57 deg.C

¹H NMR(CDCl₃)δ：3.5-3.9(14H，m)，6.48(1H，brs)，7.2-7.5(2H，m)，7.59(1H，t，J＝54Hz)，7.8-7.9(1H，m)，8.2-8.4(1H，m)

MS m/z：430(M⁺)

Preparation of example 35

2- (2-difluoromethylbenzimidazol-1-yl) -6- (3, 5-dioxopiperazin-1-yl) -4-morpholino-1, 3, 5-triazine

Melting point: 230-32 deg.C

¹H NMR(CDCl₃)δ：3.5-3.9(12H，m)，7.2-7.5(2H，m)，7.59(1H，t，J＝55Hz)，7.9-8.0(1H，m)，8.3-8.4(1H，m)

MS m/z：444(M⁺)

Preparation of example 36

Melting point: 178 ℃ C. 181 ℃ C

¹H NMR(CDCl₃)δ：3.81(2H，s)，3.5-3.9(16H，m)，7.2-7.5(7H，m)，7.52(1H，t，J＝54Hz)，7.89(1H，d，J＝8Hz)，8.30(1H，d，J＝8Hz)

MS m/z：534(M⁺)

Preparation of example 37

Melting point: 79-81 deg.C

NMR(CDCl₃)δ：2.19(3H，s)，2.60(2H，s)，3.70-4.00(16H，m)，7.30-7.50(2H，m)，7.57(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.33(1H，d，J＝8Hz)

MS m/z：472

Preparation of example 38

Melting point: 220 ℃ and 222 DEG C

NMR(CDCl₃)δ：3.80-4.00(16H，m)，6.53(1H，d，J＝2Hz)，7.01(1H，d，J＝2Hz)，7.30-7.60(4H，m)，7.80(1H，d，J＝8Hz)，8.34(1H，d，J＝8Hz)

MS m/z：510

Preparation of example 39

2- (2-Difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine (Compound 32)

Melting point: 203-

NMR(CDCl₃)δ：2.90(6H，s)，3.30-3.40(4H，m)，3.80-4.00(12H，m)，7.30-7.40(2H，m)，7.56(1H，t，J＝54Hz)，7.89(1H，d，8Hz)，8.34(1H，d，8Hz)

MS m/z：487

Preparation of example 40

Melting point: 72-75 deg.C

NMR(CDCl₃)δ：3.46(3H，s)，3.60-4.00(16H，s)，4.17(2H，s)7.30-7.50(2H，m)，7.55(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.34(1H，d，J＝8Hz)

MS m/z：488

Preparation of example 41

NMR(CDCl₃)δ：1.17(2H，m)，2.60-2.70(4H，m)，4.70(1H，brs)，3.50-4.00(16H，m)，7.30-7.50(2H，m)，7.56(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.33(1H，d，J＝8Hz)

MS m/z：474

Preparation of example 42

Melting point: 198-202 deg.C

NMR(CDCl₃)δ：1.20(3H，t，J＝7Hz)，2.42(2H，q，J＝7Hz)， 3.50-4.00(16H，m)，7.30-7.50(2H，m)，7.56(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.33(1H，d，J＝8Hz)

MS m/z：472

Preparation of example 43

2- (2-difluoromethylbenzimidazol-1-yl) -4- (4-methoxycarbonylpiperazin-1-yl) -6-morpholino-1, 3, 5-triazine

Melting point: 255 ℃ C

NMR(CDCl₃)δ：3.59(4H，brs)，3.76(3H，s)，3.70-3.95(12H，s)，7.30-7.50(2H，m)，7.55(1H，t，J＝54Hz)，7.90(1H，d，J＝8Hz)，8.34(1H，d，J＝8Hz)

MS m/z：474

The following compounds were prepared in a similar manner to that of preparation example 24.

Preparation of example 44

Melting point: 166 ℃ 168-

NMR(CDCl₃)δ：1.30(6H，s)，3.49(2H，s)，3.4-3.9(12H，m)，4.05(3H，s)，5.47(1H，s)，6.79(1H，d，J＝8Hz)，7.32(1H，t，J＝8Hz)，7.41(1H，t，J＝54Hz)，7.78(1H，d，J＝8Hz)

MS m/z：474(M⁺)

Preparation of example 45

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (cis-2, 3-dimethylmorpholino) -6-morpholinopyrimidine (Compound 37)

Melting point: 176-

NMR(CDCl₃)δ：1.20(3H，d，J＝5Hz)，1.22(3H，d，J＝5Hz)，3.6-3.7(1H，m)，3.6-4.1(13H，m)，4.05(3H，s)，5.47(1H，s)，6.79(1H，d，J＝8Hz)，7.32(1H，t，J＝8Hz)，7.42(1H，t，J＝53Hz)，7.78(1H，d，J＝8Hz)

MS m/z：474(M⁺)

Preparation of example 46

Melting point: 114-

NMR(CDCl₃)δ：1.56(3H，t，J＝7Hz)，3.49(2H，s)，3.5-3.9(12H，m)，4.32(2H，q，J＝7Hz)，5.47(1H，s)，6.78(1H，d，J＝8Hz)，7.30(1H，t，J＝8Hz)，7.41(1H，t，J＝53Hz)，7.76(1H，d，J＝8Hz)

MS m/z：488(M⁺)

Drug effectiveness determination

Next, an analysis scheme for the drug efficacy and toxicity of the heterocyclic compound represented by the general formula (I) and the results thereof will be described below. Here, the compound number of each test substance corresponds to the compound number designated for each heterocyclic compound described above.

Measurement example 1

Proliferation inhibition assay for mouse splenocytes

Splenocytes (2X 10) prepared from C57BL/6N female mice (8 weeks old, purchased from Charles River Laboratories Japan Inc.)⁶cells/mL) were suspended in RPMI1640 medium (containing 10% fetal bovine serum, 10mM HEPES, 1mM pyruvate, 4.5g/L glucose, 100 units/mL penicillin, and 0.1mg/mL streptomycin) and seeded at 0.225mL per well in wells of a 96-well plate. Serial dilutions of the test substances were added to the wells, followed by concanavalin A (Con A, 3. mu.g/mL), lipopolysaccharide (LPS, 100. mu.g/mL) or anti-mouse IgM antibody (100. mu.g/mL). Then, the cells were incubated at 37 ℃ for 3 days with 5% carbon dioxide. Then, Alamar Blue solution was added at 50. mu.L per 150. mu.L, and after 1 day of incubation, the fluorescence intensity at 590nm was measured by Cytofour 4000(Applied Biosystems) using an excitation wavelength of 530 nm. The results are shown in table 1, showing that the s-triazine analogue inhibits proliferation of mouse splenocytes induced by Con a, LPS or anti-IgM antibodies.

[ Table 1]

Test compounds	Inhibition of Con A response	Inhibition of LPS response	Inhibition of anti-IgM antibody response
				Compound 1	+	+	nd
Compound 2	+	+	+
				Compound 3	+	+	nd
Compound 4	++	++	++
				Compound 5	+	+	+
Compound 6	+	+	+
				Compound 7	+	+	+
Compound 8	++	+	++
				Compound 9	++	++	++
Compound 10	+	±	+
				Compound 12	++	++	++
Compound 13	++	++	++
				Compound 17	+	±	+
Compound 19	++	++	nd
				Compound 20	+++	+++	+++
Compound 21	+++	+++	+++
				Compound 22	+++	+++	+++
Compound 24	+	+	+
				Compound 25	++	+	nd
Compound 26	+++	++	nd
				Compound 27	+++	+	nd
Compound 28	+++	++	nd

Evaluation criteria：±：10μM＜IC50≤100uM

+：0.1μM＜IC50≤10μM

++：0.001μM＜IC50≤0.1μM

+++：IC50≤0.001μM

nd: not tested

Measurement example 2

Proliferation inhibition assay for human peripheral blood mononuclear cells

4mL of blood collected from healthy individuals was placed on 3mL of MonoPoly isolation medium, centrifuged, and a mononuclear cell (PBMC) fraction was collected. After washing with brine, PBMC (1X 10)⁵cells/mL) were suspended in RPMI1640 medium (containing 10% fetal bovine serum, 10mM HEPES, 1mM pyruvate and 4.5g/L glucose). Then, after adding an anti-CD 28 antibody (1. mu.g/mL) to the suspension, it was seeded on an anti-human CD3T cell activation plate (BD Bioscience) at a volume of 0.135mL per well. Serial dilutions of the test substance were then added to the wells and incubated at 5% carbon dioxide and 37 ℃.After 3 days, Alamar Blue solution was added at 50. mu.L per well, and after 1 day of incubation, the fluorescence intensity at 590nm was measured by Cytofour 4000(Applied Biosystems) using an excitation wavelength of 530 nm. The results are shown in table 2, showing that the s-triazine analogue inhibits growth of human T cells induced by anti-CD 3 antibody and anti-CD 28 antibody.

[ Table 2]

Measurement example 3

Activation inhibition assay for human peripheral blood mononuclear cells

PBMC (2X 10) were isolated from human peripheral blood according to the method of measurement example 2⁶cells/mL) were suspended in RPMI1640 medium containing 10% fetal bovine serum, 10mM hepes, 1mM pyruvate and 4.5g/L glucose) and seeded at 0.225mL per well in wells of a 96-well plate. Test substances were added to the wells separately, followed by anti-CD 3 antibody (2. mu.g/mL) and anti-CD 28 antibody (1. mu.g/mL). Then, after incubation for 6 hours under conditions of 5% carbon dioxide and 37 ℃ temperature, expression of CD40L and CD69, activation markers, was analyzed by flow cytometry. The result was that CD40L was expressed on 14.4% of PBMCs when cells were stimulated by both anti-CD 3 antibody and anti-CD 28 antibody, which was also CD4 positive. However, when the cells were treated with the test substance, the proportion of cells expressing CD40L decreased (ZSTK 474: 4.9%; 913: 3.7%, 1213: 3.6%). Although CD69 is expressed on 28.8% (CD 4-positive) and 50.9% (CD 4-negative) PBMCs, the proportion of CD 4-positive cells (ZSTK 474: 18.8%, 913: 10.1%, 1213: 17.7%) and CD 4-negative cells (ZSTK 474: 17.7%, 913: 10.5%, 1213: 22.4%) expressing CD69 is reduced when the cells are treated with the test substance. Thus, the s-triazine analogue was shown to inhibit lymphocyte activation (fig. 1).

Measurement example 4

Adjuvant-induced arthritis inhibition

Lyophilized Mycobacterium butyricum (Mycobacterium butyricum) suspended in Freund's incomplete adjuvant was subcutaneously administered to the caudal root of Lewis male rats (7 weeks old, purchased from Charles River Laboratories Japan Inc.) to induce adjuvant-induced arthritis. The test substance suspended in 0.5% hydroxypropyl cellulose (HPC) was then orally administered for several consecutive days 10 days after adjuvant-induced arthritis induction. In addition, after the onset of arthritis, the volume of the hind paw was measured with a hind paw edema volume meter (TK105, Physio-Tech). The results are statistically significant compared to the control group (ANOVA, Dennett test P < 0.05) for the test substances after day 14 for the model study, as shown in FIG. 2.

Measurement example 5

Collagen-induced arthritis inhibitory action

Bovine type II collagen suspended in incomplete freund's adjuvant was administered subcutaneously to the caudal roots of DBA1 male mice (7 weeks old, purchased from Charles River Laboratories Japan Inc.) on days 1 and 21. Then, from day 28 onward, when 50% of mice developed arthritis, the test substance suspended in 0.5% Hydroxypropylcellulose (HPC) was orally administered for several consecutive days. Here, effectiveness was assessed by arthritis scoring. Specifically, for each paw, the degree of arthritis was evaluated as follows: no symptoms: 0; redness or swelling of one joint: 1; redness or swelling of two or more joints: 2; redness or swelling of the entire paw: 3; and maximal redness or swelling of the entire paw: 4. the results show that ZSTK 474 inhibits the progression of collagen-induced arthritis in a dose-dependent manner. Statistically significant effects (dunnett test P < 0.05) were observed after day 30 in the groups dosed with 50 or 100mg/kg of the above-mentioned compound. However, for the group administered with 50mg/kg of the above compound, the situation worsened after day 40, and no significant difference was observed after day 44 compared with the control group (fig. 3).

Measurement example 6

Inhibition assay for rabbit synoviocytes proliferation

Rabbit synovial cells HIG-82 (4X 10) suspended in HAM medium (containing 10% fetal bovine serum, 25mM HEPES and 0.1mg/mL kanamycin)⁴cells/mL) were seeded into wells of a 96-well plate at 0.135mL per well. Then 15. mu.L of each serial dilution of the test substance was added to the wells and incubated at 5% carbon dioxide and 37 ℃. Alamar Blue solution was added at 50. mu.L per well on day 0 and day 3 and incubated for 1 day, and then the fluorescence intensity at 590nm was measured by Cytofour 4000(Applied Biosystems) using an excitation wavelength of 530 nm. As shown in table 3, the s-triazine analogue was demonstrated to inhibit cell division of rabbit synoviocytes.

[ Table 3]

Measurement example 7

Allogenic mixed lymphocyte reaction

Splenocytes prepared from C57BL/6N female mice (8-10 weeks old, purchased from Charles River Laboratories Japan Inc.) and peripheral lymph node monocytes prepared from BALB/C female mice (8-10 weeks old, purchased from Charles River Laboratories Japan Inc.) were used as the stimulator and responder cells, respectively. The cells were suspended in RPMI1640 medium (containing 10% fetal bovine serum, 100 units/mL penicillin and 0.1mg/mL streptomycin), respectively (2X 10)⁶cells/mL). Then, stimulated cells (50. mu.L) treated with mitomycin C (50mg/mL, 30 min) were added to the responsive cells (100. mu.L). Then adding into the pores respectivelySerial dilutions of the test substances were incubated for 86 hours at 37 ℃ in 5% carbon dioxide. Finally, cell proliferation was detected using the BrdU cell proliferation kit (Calbiochem). As a result, as shown in Table 4, it was found that the s-triazine analogue inhibited the allogeneic mixed lymphocyte reaction.

[ Table 4]

Measurement example 8

Proliferation inhibitory Activity on blood tumor cells

Daudi cells (5X 10) suspended in RPMI1640 medium (containing 10% fetal bovine serum, 10mM HEPES, 1mM pyruvate, 4.5g/L glucose, 100 units/mL penicillin and 0.1mg/mL streptomycin)⁵cells/mL), Jurkat cells (5X 10)⁵cell/mL), THP-1 cell (5X 10)⁵cell/mL), U937 cells (5X 10)⁵cell/mL) and HL 60 cells (5X 10)⁵cells/mL) were seeded in wells of a 96-well plate at a capacity of 0.135mL per well. Serial dilutions of the test substance were added to wells at 15 μ L per well and incubated for 3 days at 5% carbon dioxide and 37 ℃. Then 50. mu.L of Alamar Blue solution was added, and after 1 day of incubation, the fluorescence intensity at 590nm was measured by Cytofour 4000(Applied Biosystems) using an excitation wavelength of 530 nm. As shown in table 5, the results show that the s-triazine analogue inhibits the proliferation of hematologic tumor cells.

[ Table 5]

Nd: not tested.

Measurement example 9

Therapeutic effect on human B lymphoma xenograft model

Daudi cells (1X 10) cultured in RPMI1640 medium (containing 10% fetal calf serum, 10mM HEPES, 1mM pyruvate, 4.5g/L glucose, 100 units/mL penicillin and 0.1mg/mL streptomycin)⁷cells/mL) were implanted subcutaneously into the chest of 8-week-old NOD/SCID mice. After day 20, when the tumor had grown to about 800mm³At volume (b), compound 8(400mg/kg) was administered orally. The results are shown in fig. 4, where the increase in tumor volume was suppressed in this model.

Measurement example 10

Fulminant hepatitis model

BALB/c male mice (7 weeks old, purchased from Charles River Laboratories Japan Inc.) were used in this experiment. After oral administration of the test substance suspended in 5% HPC, galactosamine (800mg/kg) and LPS (110. mu.g/kg) were administered intraperitoneally. Then, the survival rate was obtained 72 hours after the administration. As shown in Table 6, the test substances improved the lethal effect caused by galactosamine and LPS.

[ Table 6]

Measurement example 11

Toxicity testing of single oral administration

A single oral dose toxicity of typical heterocyclic compounds was examined with SD male rats (6 weeks of age, body weight 162-188g) with the result that no instances of death were observed even at 1200mg/kg for compound 8 and LD for compound 14₅₀600-900 mg/kg.

Measurement example 12

Ames test

The test substances (compound 8, compound 10, compound 12 and compound 14) were tested for mutagenesis using 5 strains of Salmonella typhimurium TA98, TA100, TA1535, TA1537 and E.coli WP2uvrA according to the preincubation method. As a result, no increase in the number of colonies of back mutation was observed in any of the tested strains, regardless of the presence or absence of metabolic activation due to S-9, even in 5000. mu.g/plate (maximum dose), so that the mutagenesis effect was negative. Thus, in addition to in vivo tests, the s-triazine analogs were found to be safer compounds as measured by rat adjuvant-induced arthritis, mouse collagen-induced arthritis, human lymphoma type B xenograft models, fulminant hepatitis inhibition assays, and single oral toxicity tests.

The agent of the present invention inhibits the responses to T cells and B cells induced by Con a, LPS, anti-IgM antibody, anti-CD 3 antibody + anti-CD 28 antibody, and thus shows an inhibitory effect on immune cells PI 3K. In particular, the medicaments of the invention are useful for the treatment and prevention of immune system disorders due to hyperactivity of PI 3K. Immune system disorders due to hyperactivity of PI3K may be: autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome; organ dysfunction associated with autoimmune diseases such as uveitis, glomerulonephritis, thyroiditis, pancreatitis, bone destruction, etc.; rejection after tissue transplantation, graft versus host disease after bone marrow transplantation; inflammatory bowel disease such as ulcerative colitis or crohn's disease; inflammatory or allergic skin diseases such as psoriasis or atopic dermatitis; inflammatory or allergic respiratory disorders such as chronic obstructive pulmonary disease or asthma; allergic conjunctivitis or rhinitis; hematological tumors derived from immune cells such as B cell lymphoma, T cell lymphoma, myeloid leukemia; sepsis triggered by gram negative bacterial or coronavirus infection, severe acute respiratory syndrome, fulminant hepatitis, and the like.

Measurement example 13

Test for measuring blood levels

Pharmacokinetic studies were performed with 6 week old BDF1 male mice. The test substance was mixed with hydroxypropyl cellulose (low molecular weight form) [ hpc (l) ] 2.5 times the weight of the drug, and dissolved in dichloromethane. The solvent was evaporated to dryness. To prepare the administration form, the residue was suspended in distilled water to prepare a drug level of 20 mg/mL. The test compound was administered orally to mice starved for 16 hours at a dose of 200 mg/kg. Blood was collected from the orbit of two mice 1 hour after the administration to obtain serum. To 100. mu.L of the obtained serum were added an internal standard solution and 1ml of distilled water, followed by extraction with diethyl ether. The solvent was evaporated in vacuo and the residue was dissolved in the eluent to prepare the sample for HPLC assay. HPLC was performed using a reverse phase type column, and acetonitrile-phosphate buffer (pH 2.5) was used as a mobile phase. Drug levels in sample sera were calculated using regression curves from standards (Y ═ aX + b). The results are shown in Table 7 below.

[ Table 7]

As shown in the above test results, the compound of the present invention having an acyl group at the 4-position of the piperazine ring rapidly showed high blood levels 1 hour after administration, compared to the known control compounds 2, 3 and 9.

Measurement example 14

Proliferation inhibitory Activity on solid tumor cells

MCF-7 cells used in this assay were established from human breast cancer and were incubated at 37 ℃ and 5% CO₂Under the conditions of (1), conventional culture was carried out in MEM medium supplemented with 10% fetal bovine serum, 25mM HEPES and 0.1mg/ml kanamycin. MCF-7 cells in logarithmic growth phase were treated with insulin/EDTA to prepare single cell suspensions adjusted to 4.0X 10 in MEM medium (supplemented with 10% fetal bovine serum, 25mM HEPES and 0.1mg/ml kanamycin)⁴Cells/ml. Test compounds were dissolved in DMSO and diluted to 2.0X 10 with RPMI1640 medium (supplemented with 10% fetal bovine serum, 25mM HEPES and 0.1mg/ml kanamycin)^-4To 2.0X 10^-9The concentration of M.

The cell suspension was injected into a 96-well microplate at 0.1ml per well and cultured for 24 hours to attach the cells to the microplate. Then 0.1ml of the sample solution was added and incubated at 37 ℃ and 5% COx for 72 hours.

The 50% growth inhibitory concentration (GI) was calculated from the growth inhibition at different sample concentrations₅₀μ M). The results are shown in Table 8.

In the foregoing, when cells other than MCF-7 were used, the following medium was used instead of adding 10% fetal bovine serum, and the following single cell suspension was prepared.

PC-3 prostate cancer cells: 10% fetal bovine serum in F12K Medium, 2X 10⁴A single cell suspension of cells;

a549 lung cancer cell: 10% fetal bovine serum in DMEM medium, 1.5X 10⁴A single cell suspension of cells;

WiDr colon cancer cells: 10% fetal bovine serum in MEM Medium, 3X 10⁴Of cellsA single cell suspension;

B16F10 melanoma cells RPMI1640 culture medium 10% fetal bovine serum, 1X 10⁴Single cell suspensions of cells.

[ Table 8]

GI₅₀(μM)

	PC-3	B16F10	WiDr	A549	MCF-7
						Compound 25	0.27	3.21	0.92	0.99	0.98
Compound 27	0.31	4.63	1.32	1.12	1.14
						Compound 28	0.27	2.66	1.27	0.83	1.21
Compound 29	0.32	3.93	2.58	1.32	0.70
						Compound 30	0.77	3.32	3.74	1.52	0.41
Compound 31	0.32	1.51	1.43	0.65	0.05
						Compound 32	0.38	2.14	1.33	0.83	＜0.04
Compound 33	0.51	2.91	1.46	1.05	＜0.04
						Compound 34	0.55	2.91	2.07	1.38	0.14
Compound 35	0.73	2.84	2.10	1.22	0.10

INDUSTRIAL APPLICABILITY

The agents of the invention are useful in the prevention or treatment of disorders of the immune system in which PI3K is involved, such as autoimmune diseases, organ transplantation, allergic or inflammatory diseases, hematological neoplasms, sepsis, and the like. In addition, they are useful for the treatment of solid tumors. In addition, they may be used as PI3K inhibitors for the treatment of various disorders.

Claims

1. An immunosuppressant comprising a heterocyclic compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient:

wherein the content of the first and second substances,

x represents a nitrogen atom or CH;

R₁and R₂Two or any one of them represents a hydrogen atom, a hydroxyl group, a halogenAmino group, C₁-C₆Alkylamino radical, C₁-C₆Alkoxy radical, C₁-C₆An alkyl group or a cyano group;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

R₆representing one or two C₁-C₆Morpholino optionally substituted with an alkyl group; by hydroxy radicals C₁-C₆Pyrrolidinyl optionally substituted with alkyl; by one or two oxygen atoms, hydroxy groups, formyl groups or C₁-C₆Alkyl optionally substituted piperidino; optionally substituted by one or two oxygen atoms, and the nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆A piperazinyl group optionally substituted with a substituent of an oxoalkyl group, an aromatic carbonyl group, a benzylcarbonyl group and a substituted carbamoyl group; or optionally substituted by one or two oxygen atoms, and the nitrogen in position 4 being selected from formyl, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆1, 4-diazepinyl optionally substituted with the substituents of oxoalkyl, aromatic carbonyl, benzylcarbonyl and substituted carbamoyl.

2. The immunosuppressant of claim 1, wherein R₁Or R₂Any of which is a hydroxyl group.

3. The immunosuppressant of claim 1 or 2, wherein R₁Or R₂Any of which is a hydroxyl group, and R₃Is difluoromethyl.

4. The immunosuppressant of claim 1, wherein R₁And R₂Are all hydrogen, and R₃Is difluoromethyl.

5. The immunosuppressant of any one of claims 1-4, wherein R₆Is 4-acetyl piperazine.

6. The immunosuppressant of claim 1, wherein the heterocyclic compound is selected from the group consisting of:

2- (2-methylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (piperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (3-oxopiperazin-1-yl) -1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -6- (4-formylpiperazin-1-yl) -4-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -6- (3, 5-dioxopiperazin-1-yl) -4-morpholino-1, 3, 5-triazine;

2- (benzimidazol-1-yl) -4- (trans-2, 3-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6-piperidino-1, 3, 5-triazine;

2- (6-amino-2-difluoromethylbenzimidazol-1-yl) -4- (cis-2, 3-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-6-ethoxybenzoimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-4-methylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-5-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-aminobenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (6-amino-2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholino-1, 3, 5-triazine;

2- (2-methylbenzimidazol-1-yl) -4, 6-dimorpholinopyrimidine;

2- (2-hydroxymethylbenzoimidazol-1-yl) -4, 6-dimorpholinopyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (piperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (5-oxo-1, 4-diazepin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -4-morpholino-6- (3-oxopiperazin-1-yl) pyrimidine;

2- (2-difluoromethylbenzimidazol-1-yl) -6- (4-formylpiperazin-1-yl) -4-morpholinopyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholinopyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (3, 3-dimethylmorpholino) -6-morpholinopyrimidine;

2- (2-aminobenzimidazol-1-yl) -4, 6-dimorpholinopyrimidine;

2- (2-difluoromethyl-4-methoxybenzimidazol-1-yl) -4- (2, 6-dimethylmorpholino) -6-morpholinopyrimidine;

2- (5-amino-2-difluoromethylbenzimidazol-1-yl) -4, 6-bis (2, 6-dimethylmorpholino) pyrimidine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -6-morpholinopyrimidine;

2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholino-1, 3, 5-triazine;

2- (6-amino-4-chloro-2-difluoromethylbenzimidazol-1-yl) -4- [ methyl (1-methylpiperidin-4-yl) amino ] -6-morpholino-1, 3, 5-triazine;

2- (4-chloro-2-difluoromethyl-5-hydroxybenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (5, 6-dimethyl-2-difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (4-amino-2-difluoromethylbenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- [4- (N, N-dimethylcarbamoyl) piperazin-1-yl ] -6-morpholinopyrimidine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -6-morpholino) -1, 3, 5-triazine;

4- (4-acetylmethylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6- (cis 2, 3-dimethylmorpholino) pyrimidine;

7. The immunosuppressant of claim 1, wherein the heterocyclic compound is selected from the group consisting of:

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2-hydroxymethylpyrrolidin-1-yl) -morpholino-1, 3, 5-triazine;

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (2, 2-dimethylmorpholino) -6-morpholinopyrimidine; and

2- (2-difluoromethyl-4-hydroxybenzimidazol-1-yl) -4- (3, 3-dimethylmorpholino) -6-morpholinopyrimidine.

8. The immunosuppressant of claim 1, wherein the heterocyclic compound is selected from the group consisting of:

2- (2-difluoromethylbenzimidazol-1-yl) -4, 6-dimorpholino-1, 3, 5-triazine;

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholinopyrimidine; and

4- (4-acetylpiperazin-1-yl) -2- (2-difluoromethylbenzimidazol-1-yl) -6-morpholino-1, 3, 5-triazine.

9. The immunosuppressant of any one of claims 1-8, wherein the condition of interest is rejection or graft-versus-host disease; inflammatory bowel disease; inflammatory or allergic skin diseases; inflammatory or allergic respiratory disorders; (ii) an autoimmune disease; malignant lymphoma, multiple myeloma, leukemia; sepsis, fulminant hepatitis.

10. The immunosuppressant of claim 9, wherein the inflammatory bowel disease is ulcerative colitis or crohn's disease.

11. The immunosuppressant according to claim 9, wherein the inflammatory or allergic skin disease is psoriasis or atopic dermatitis.

12. The immunosuppressant of claim 9 wherein the inflammatory or allergic respiratory disorder is obstructive pulmonary disease or asthma.

13. The immunosuppressant of claim 9, wherein the autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or sjogren's syndrome.

14. A heterocyclic compound represented by the general formula (II):

wherein the content of the first and second substances,

n represents 0 to 2;

x represents a nitrogen atom or CH;

y represents- (CH)₂)n₁-, in which n₁Is 1 to 2;

R₄and R₅Represents a hydrogen atom or C₁-C₆An alkyl group;

R₇represents a hydrogen atom, C₁-C₆Alkyl radical, formyl radical, C₁-C₆Hydroxyalkyl radical, C₁-C₆Alkoxycarbonyl group, C₁-C₆Oxoalkyl, aromatic carbonyl, benzylcarbonyl or substituted carbamoyl, with the proviso that when n ═ 0 and n₁When 1, a hydrogen atom or C₁-C₆Alkyl groups are excluded.

15. The compound of claim 14, wherein the heterocyclic compound is:

16. An antitumor agent comprising the compound according to claim 14 or 15 as an active ingredient.