JPH10101671A - Nitric oxide synthase inhibitor - Google Patents

Abstract

(57) [Summary] (Modified) [PROBLEMS] To provide a nitric oxide synthase inhibitor. SOLUTION: General formula (1) [R ¹ is a hydrogen atom, a substituent or an unsubstituted lower alkyl group, a cycloalkyl group, etc., R ² is a naphthyl group, a cycloalkyl group, a furyl group, a thienyl group, etc., and R ³ is a hydrogen atom, a phenyl group or a lower alkyl group. A group, R ⁴ is a hydrogen atom,
Lower alkyl group, lower alkoxycarbonyl group, a phenyl-lower alkyl group, R ⁵ is a hydrogen atom or a lower alkyl group, R ⁶ represents a hydrogen atom, a lower alkyl group, a phenyl-lower alkyl group or a substituted benzoyl group, R ¹ and R ⁵ may combine with each other to form a lower alkylene group;
Represents a carbonyl group or a sulfonyl group, A represents a single bond, a lower alkylene group or a lower alkenylene group, and n represents 0 or 1]. Inhibition of nitric oxide synthase containing a pyrazolo [1,5-a] pyrimidine derivative as an active ingredient Agent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a novel inhibitor of NO (nitric oxide) synthase, and more particularly to an agent which inhibits the induction of inducible NO synthase.

[0002]

2. Description of the Related Art In the early 1980s, during the study of nitrogen oxides in vivo, it was first discovered that NO (nitric oxide) was produced in vivo. Following that discovery, NO attracted the attention of many researchers, and in 1987, it was reported that NO was the main body of vascular endothelium-derived relaxing factor. At present, the relationship between NO physiological functions and disease states has been elucidated in a wide range of fields such as circulation, immunity, and nervous system.

Among them, it has been elucidated that, for example, NO constantly produced in the body plays an important role in maintaining circulatory dynamic homeostasis. On the other hand, in sepsis, it is said that a large amount of NO is produced by the action of cytokines activated by endotoxin, and this causes an endotoxin shock state such as endothelial cell damage and reduced myocardial contractility.

[0004] NO is produced from L-arginine by NO synthase (NOS). The enzymes are mainly divided into inducible NO related to NO production during disease states.
S (iNOS) and constitutive NOS (c
NOS).

[0005] As described above, NO is involved in various diseases such as sepsis. Therefore, NO has been studied for the purpose of elucidating its mechanism and, consequently, applying it to a therapeutic drug for those diseases.
Research on S inhibitors has been advanced, and typical examples thereof include arginine analogs such as N-ω-nitro-L-arginine.

[0006] However, most of the known NOS inhibitors including the above representative examples inhibit not only iNOS but also cNOS. Even the regulation of circulatory dynamics is suppressed, and it is not possible to avoid side effects such as an increase in blood pressure and a decrease in organ blood flow. In addition, there is a concern that the use of these methods may affect the central nervous system and impotence.

[0007] As described above, conventionally known NOS inhibitors cannot be evaluated as pharmaceuticals, and instead, a new substance capable of selectively inhibiting iNOS has been provided in the art. Requested.

[0008]

Accordingly, an object of the present invention is to provide a substance which can selectively inhibit only iNOS and a nitric oxide synthase inhibitor using the same, which is required in the art. It is in.

The research group of the present inventors has long been engaged in the synthesis of various compounds and the study and elucidation of their pharmacological actions for the purpose of developing active compounds for pharmaceutical preparations. Has successfully synthesized a series of pyrazolopyrimidine derivatives having a powerful analgesic effect and filed a patent application for an invention relating to this compound [WO95.
/ 35298].

[0010] In subsequent studies, the present inventors have determined by the fact that the above-mentioned series of compounds has an inhibitory effect on the induction of iNOS, separately and independently of the analgesic effect previously found. The present invention has been newly found, and the present invention has been completed.

[0011]

That is, according to the present invention,
Pyrazolo [1,5-a] represented by the following general formula (1)
A nitric oxide synthase inhibitor comprising a pyrimidine derivative as an active ingredient is provided.

[0012]

Embedded image

In the above formula (1), R ¹ is a hydrogen atom, a lower alkyl group which may have a thienyl group, a lower alkoxy group, a lower alkylthio group, an oxo group or a hydroxyl group as a substituent, a cycloalkyl group, a thienyl group. Group,
R ² represents a naphthyl group, a cycloalkyl group, a furyl group, a lower alkenyl group or a phenyl group which may have 1 to 3 groups selected from a lower alkyl group, a lower alkoxy group, a phenylthio group and a halogen atom as a substituent. , A furyl group, a thienyl group, a pyridyl group which may be substituted with a halogen atom, a phenoxy group which may be substituted with a halogen atom or a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, or a halogen substitution. One of groups selected from a lower alkyl group, a halogen-substituted lower alkoxy group, a lower alkoxycarbonyl group, a hydroxyl group, a phenyl lower alkoxy group, an amino group, a cyano group, a lower alkanoyloxy group, a phenyl group and a di-lower alkoxyphosphoryl lower alkyl group; A phenyl group which may have up to 3 , R ³ is a hydrogen atom, a phenyl group or a lower alkyl group, R ⁴ is a hydrogen atom, a lower alkyl group,
A lower alkoxycarbonyl group, a phenyl lower alkyl group, a phenyl group or a halogen atom which may have a phenylthio group as a substituent, R ⁵ is a hydrogen atom or a lower alkyl group, R ⁶ is a hydrogen atom, a lower alkyl group, a phenyl lower group; Alkyl or a benzoyl group having 1 to 3 groups selected from a lower alkoxy group, a halogen-substituted lower alkyl group and a halogen atom as a substituent, and R ¹ and R ⁵ are bonded to each other to form a lower alkylene group. Q may be a carbonyl group or a sulfonyl group,
A represents a single bond, a lower alkylene group or a lower alkenylene group, and n represents 0 or 1.

The derivative represented by the above general formula (1) is
Has an inhibitory action on nitric oxide synthesis, in particular, an action of selectively inhibiting inducible nitric oxide synthase (iNOS),
Therefore, it is characterized in that it hardly exhibits side effects such as increased blood pressure, decreased organ blood flow, and adverse effects on the central nervous system.

Examples of each group in the above general formula (1) representing the active ingredient of the nitric oxide synthase inhibitor of the present invention include the following groups. That is, examples of the lower alkyl group include linear or branched lower alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, and hexyl groups.

Examples of the cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
Examples include cycloheptyl and cyclooctyl groups.

Examples of the lower alkoxy group include methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentyloxy, hexyloxy and the like.

Examples of the lower alkylthio group include a methylthio, ethylthio, propylthio, butylthio, pentylthio, hexylthio group and the like.

The halogen atom includes fluorine, chlorine, bromine and iodine atoms.

Examples of the halogen-substituted lower alkyl group include trifluoromethyl, pentafluoroethyl, heptafluoropropyl, nonafluorobutyl, undecafluoropentyl, and tridecafluorohexyl.

The halogen-substituted lower alkoxy group includes
Examples thereof include trifluoromethoxy, pentafluoroethoxy, heptafluoropropoxy, nonafluorobutoxy, undecafluoropentyloxy, and tridecafluorohexyloxy groups.

Examples of the lower alkoxycarbonyl group include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl and the like.

Examples of the di-lower alkoxyphosphoryl lower alkyl group include dimethoxyphosphorylmethyl, diethoxyphosphorylmethyl, dipropoxyphosphorylmethyl, diisopropoxyphosphorylmethyl, dibutoxyphosphorylmethyl, dipentyloxyphosphorylmethyl, dihexyloxyphosphorylmethyl, (Dimethoxyphosphoryl) ethyl, 2- (diethoxyphosphoryl) ethyl,
Examples thereof include a 3- (diethoxyphosphoryl) propyl group.

The naphthyl group includes 1-naphthyl and 2-naphthyl groups.

Examples of the lower alkylene group include a methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene group and the like.

As the lower alkenylene group, vinylene,
Examples thereof include a propenylene group.

Examples of the pyridyl group which may be substituted with a halogen atom include 2-pyridyl, 3-pyridyl, 4-pyridyl, 6-chloro-2-pyridyl, 5-chloro-2-
Pyridyl, 4-chloro-2-pyridyl, 3-chloro-2
-Pyridyl, 6-chloro-3-pyridyl, 5-chloro-
3-pyridyl, 4-chloro-3-pyridyl, 2-chloro-3-pyridyl, 2-chloro-4-pyridyl, 3-chloro-4-pyridyl, 6-fluoro-3-pyridyl, 6-
Examples thereof include a bromo-3-pyridyl group and a 6-iodo-3-pyridyl group.

Examples of the phenoxy group which may be substituted by a halogen atom include phenoxy, 2-chlorophenoxy, 3-chlorophenoxy, 4-chlorophenoxy,
-Fluorophenoxy, 4-bromophenoxy, 4-iodophenoxy and the like.

The thienyl group includes 2-thienyl and 3-thienyl groups, and the furyl group includes 2-furyl and 3-furyl groups.

The lower alkenyl group includes vinyl, allyl, isopropenyl, 1-butenyl, 2-butenyl, 3
-Butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl group and the like.

Examples of the phenyl lower alkyl group include a benzyl, 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl, 4-phenylbutyl, 5-phenylpentyl, 6-phenylhexyl group and the like.

Examples of the phenyl lower alkoxy group include benzyloxy, 2-phenylethoxy, 3-phenylpropoxy, 4-phenylbutoxy, 5-phenylpentyloxy, 6-phenylhexyloxy and the like.

Examples of the lower alkanoyloxy group include acetoxy, propionyloxy, butyryloxy, valeryloxy, pivaloyloxy, hexanoyloxy, heptanoyloxy and the like.

The lower alkyl group which may have a thienyl group, a lower alkoxy group, a lower alkylthio group, an oxo group or a hydroxyl group as a substituent includes 2-thienylmethyl, 3-thienylmethyl,
-Thienylmethyl, 1- (2-thienyl) ethyl, 1-
(3-thienyl) ethyl, 2- (2-thienyl) ethyl, 2- (3-thienyl) ethyl, 3- (2-thienyl) propyl, 4- (2-thienyl) butyl, 5- (2
-Thienyl) pentyl, 6- (2-thienyl) hexyl, methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, pentyloxymethyl, hexyloxymethyl, 1-methoxyethyl, 2-methoxyethyl, 3-methoxypropyl, -Methoxybutyl, 5-
Methoxypentyl, 6-methoxyhexyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 3-hydroxybutyl,
4-hydroxypentyl, 5-hydroxyhexyl, methylthiomethyl, ethylthiomethyl, propylthiomethyl, butylthiomethyl, pentylthiomethyl, hexylthiomethyl, 2-methylthioethyl, 3-methylthiopropyl, 4-methylthiobutyl, 5- Methylthiopentyl, 6-methylthiohexyl, formyl, formylmethyl, acetyl, 2-formylethyl, 2-oxopropyl, propionyl, 3-formylpropyl, 3-oxobutyl, 2-oxobutyl, butyryl, 4-formylbutyl, 4-oxo Pentyl, 3-oxopentyl, 2-
Oxopentyl, valeryl, 5-formylpentyl, 5
Examples thereof include -oxohexyl, 4-oxohexyl, 3-oxohexyl, 2-oxohexyl, and a hexanoyl group.

The phenyl group which may have 1 to 3 groups selected from a lower alkyl group, a lower alkoxy group, a phenylthio group and a halogen atom as a substituent includes:
Phenyl, 2-methylphenyl, 3-methylphenyl,
4-methylphenyl, 4-ethylphenyl, 4-propylphenyl, 4-butylphenyl, 4-t-butylphenyl, 4-pentylphenyl, 4-hexylphenyl,
2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl, 2,6-dimethylphenyl, 3,4-dimethylphenyl, 3,5-dimethylphenyl, 2-methoxyphenyl, 3-methoxy Phenyl, 4-methoxyphenyl, 4-ethoxyphenyl, 4
-Propoxyphenyl, 4-butoxyphenyl, 4-pentyloxyphenyl, 4-hexyloxyphenyl,
2,3-dimethoxyphenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3,4,5-trimethoxy Phenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-iodophenyl, 4-fluorophenyl, 4- (phenylthio) phenyl, 3- (phenylthio) phenyl, 2- (phenylthio) phenyl group Etc. can be exemplified.

As a substituent, a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, a halogen-substituted lower alkyl group, a halogen-substituted lower alkoxy group, a lower alkoxycarbonyl group, a hydroxyl group, a phenyl-lower alkoxy group, an amino group, a cyano group Examples of the phenyl group which may have 1 to 3 groups selected from a lower alkanoyloxy group, a phenyl group and a di-lower alkoxyphosphoryl lower alkyl group include the following groups.

That is, phenyl, 2-methylphenyl, 3
-Methylphenyl, 4-methylphenyl, 4-ethylphenyl, 4-propylphenyl, 4-butylphenyl,
4-t-butylphenyl, 4-pentylphenyl, 4-
Hexylphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 4-ethoxyphenyl, 4-propoxyphenyl, 4-butoxyphenyl, 4-pentyloxyphenyl, 4-hexyloxyphenyl, 2,3-dimethoxy Phenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl, 2,6
-Dimethoxyphenyl, 3,4-dimethoxyphenyl,
3,5-dimethoxyphenyl, 2,3,4-trimethoxyphenyl, 2,3,5-trimethoxyphenyl, 2,
3,6-trimethoxyphenyl, 2,4,5-trimethoxyphenyl, 2,4,6-trimethoxyphenyl,
3,4,5-trimethoxyphenyl, 3,4,5-triethoxyphenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl, 2-bromo Phenyl, 3-bromophenyl, 4-bromophenyl, 4-iodophenyl, 2,3-dichlorophenyl,
2,4-dichlorophenyl, 2-nitrophenyl, 3-
Nitrophenyl, 4-nitrophenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl,
4-trifluoromethylphenyl, 4-pentafluoroethylphenyl, 4-heptafluoropropylphenyl, 4-nonafluorobutylphenyl, 4-undecafluoropentylphenyl, 4-tridecafluorohexylphenyl, 2-methoxycarbonylphenyl, 3-methoxycarbonylphenyl, 4-methoxycarbonylphenyl, 4-ethoxycarbonylphenyl, 4-propoxycarbonylphenyl, 4-butoxycarbonylphenyl, 4-pentyloxycarbonylphenyl, 4-hexyloxycarbonylphenyl, 2-biphenyl,
-Biphenyl, 4-biphenyl, 2- (diethoxyphosphorylmethyl) phenyl, 3- (diethoxyphosphorylmethyl) phenyl, 4- (diethoxyphosphorylmethyl) phenyl, 4- (dimethoxyphosphorylmethyl) phenyl, 4- (dimethoxyphosphorylmethyl) phenyl Isopropoxyphosphorylmethyl) phenyl, 3,5-dimethoxy-4-ethoxyphenyl,
3,5-dimethoxy-4-propoxyphenyl, 4-butoxy-3,5-dimethoxyphenyl, 3,5-dimethoxy-4-pentyloxyphenyl, 3,5-dimethoxy-4-hexyloxyphenyl, 2,3- Bis (trifluoromethyl) phenyl, 2,4-bis (trifluoromethyl) phenyl, 2,5-bis (trifluoromethyl) phenyl, 2,6-bis (trifluoromethyl) phenyl, 3,4-bis ( Trifluoromethyl) phenyl, 3,5-bis (trifluoromethyl) phenyl,
3,5-dimethoxy-4-hydroxyphenyl, 3,5
-Diethoxy-4-hydroxyphenyl, 3,5-dipropoxy-4-hydroxyphenyl, 4-benzyloxy-3,5-dimethoxyphenyl, 4-benzyloxy-3,5-diethoxyphenyl, 3,5-dimethoxy-
4- (2-phenylethoxy) phenyl, 4-acetoxy-3,5-dimethoxyphenyl, 3,5-dimethoxy-4-propionyloxyphenyl, 2-chloro-3,
5-dimethoxyphenyl, 4-chloro-3,5-dimethoxyphenyl, 4-bromo-3,5-dimethoxyphenyl, 3,5-dimethoxy-4-iodophenyl, 3,5
-Dichloro-4-methoxyphenyl, 3,5-dichloro-4-ethoxyphenyl, 2-aminophenyl, 3-aminophenyl, 4-aminophenyl, 2-cyanophenyl, 3-cyanophenyl, 4-cyanophenyl, -Trifluoromethoxyphenyl, 3-trifluoromethoxyphenyl, 2-trifluoromethoxyphenyl, 4-
Pentafluoroethoxyphenyl, 4-heptafluoropropoxyphenyl, 4-nonafluorobutoxyphenyl, 4-undecafluoropentyloxyphenyl, 4
-Tridecafluorohexyloxyphenyl, 3,5-
Bis (trifluoromethoxy) phenyl, 3,4,5-
A tris (trifluoromethoxy) phenyl group and the like can be exemplified.

The phenyl group which may have a phenylthio group as a substituent includes phenyl, 4- (phenylthio) phenyl, 3- (phenylthio) phenyl, 2- (phenylthio) phenyl,
A (phenylthio) phenyl group can be exemplified.

The benzoyl group having 1 to 3 groups selected from a lower alkoxy group, a halogen-substituted lower alkyl group and a halogen atom as a substituent includes 2-chlorobenzoyl, 3-chlorobenzoyl, 4-chlorobenzoyl, -Fluorobenzoyl, 2-bromobenzoyl,
2-iodobenzoyl, 2,4-dichlorobenzoyl,
3,4-dichlorobenzoyl, 2,5-dichlorobenzoyl, 2,6-dichlorobenzoyl, 2-trifluoromethylbenzoyl, 3-trifluoromethylbenzoyl, 4-trifluoromethylbenzoyl, 3,5-bis (trifluoro Methyl) benzoyl, 3,4,5-tris (trifluoromethyl) benzoyl, 2-methoxybenzoyl, 3-methoxybenzoyl, 4-methoxybenzoyl, 2,3-dimethoxybenzoyl, 2,4-dimethoxybenzoyl, 3,5 -Dimethoxybenzoyl,
Examples thereof include 3,4,5-trimethoxybenzoyl, 2-ethoxybenzoyl, 2-propoxybenzoyl, 2-butoxybenzoyl, 2-pentyloxybenzoyl, and 2-hexyloxybenzoyl groups.

The pyrazolo [1,5-a] pyrimidine derivative represented by the general formula (1) is a nitric oxide synthase inhibitor, particularly an inducible nitric oxide synthase (iNOS).
As a drug that selectively inhibits, it is useful for the treatment and prevention of sepsis, endotoxin shock, rheumatoid arthritis, etc., and has the advantage of having very few side effects as seen in conventional nitric oxide synthase inhibitors.

As the pyrazolo [1,5-a] pyrimidine derivative preferable as the nitric oxide synthase inhibitor, in the above general formula (1), R ⁵ and R ⁶ are each a hydrogen atom, and Q is a carbonyl group. , A is a single bond and n is 0. Among the said compounds, in particular with a lower alkyl group in which R ¹ which may have a hydroxyl group or a lower alkoxy group as a phenyl group or a substituted group, R ²
Is a phenyl group having 1 to 3 groups selected from a lower alkoxy group, a halogen-substituted lower alkyl group and a halogen atom as a substituent, wherein R ⁴ is a hydrogen atom or a phenyl group, more specifically, R ¹ Is a phenyl, methyl, ethyl, n-butyl or n-pentyl group, and R
² is a 4-ethoxy-3,5-dimethoxyphenyl group,
Compounds that are 3,4,5-trimethoxyphenyl, 2-methoxyphenyl, 2,4-dichlorophenyl or 2-trifluoromethylphenyl are preferred.

Specific examples of preferred pyrazolo [1,5-a] pyrimidine derivatives include, for example, 5-n-butyl-
7- (3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine, 5-phenyl-7-
(3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine and 5-n-butyl-7-
(2-trifluoromethylbenzoylamino) pyrazolo [1,5-a] pyrimidine can be exemplified, and among these, 5-n-butyl-7- (3,4,5-trimethoxybenzoylamino) pyrazolo [ [1,5-a] pyrimidine is optimal.

The active ingredient compound of the present invention represented by the general formula (1) can be produced by various methods.
As a specific example, the method described in the above-mentioned WO 95/35298 can be exemplified.

Typically, the process involves the condensation of an appropriate carboxylic acid ester with 3-aminopyrazolyl to give 7-hydroxypyrazolo [1,5-a] pyrimidines, which are then It can be carried out by halogenating to 7-halogenopyrazolo [1,5-a] pyrimidines, further treating them with aqueous ammonia or hydrazine to convert them into 7-amino forms, and reacting them with acid halides.

Specific examples of the active ingredient compound of the present invention obtained according to such a method are shown in Tables 1 to 5 below and are described in Example No. Each compound shown as 1-134 can be illustrated.

The compound represented by the general formula (1) is
It can be a pharmaceutically acceptable acid addition salt, and these salts are also included in the active ingredient compound of the nitric oxide synthase inhibitor of the present invention. Examples of the acid capable of forming the acid addition salt include inorganic acids such as hydrochloric acid, hydrobromic acid, and sulfuric acid, and organic acids such as oxalic acid, fumaric acid, maleic acid, tartaric acid, and citric acid. The salt formation reaction can be performed according to a conventional method.

In the compounds represented by the above general formula (1), those in which R6 is a hydrogen atom can be converted into an alkali metal salt such as a sodium salt, a potassium salt or the like in accordance with a conventional method. Calcium salts, magnesium salts and the like, and other copper salts and the like can be used, and these salts are also included in the active ingredient compound of the nitric oxide synthase inhibitor of the present invention.

In the compound represented by the general formula (1),
Some of the compounds in which A is an alkenylene group and some of the compounds in which R1 is a lower alkenyl group can have cis and trans isomer structures, and the nitric oxide synthase inhibitor of the present invention is effective in both cases. It can be a component.

Some of the compounds represented by the general formula (1) have optical isomers having an asymmetric center at a carbon atom. Both the active form and the racemic form can be used as the active ingredient.

The nitric oxide synthase inhibitor of the present invention comprises a compound represented by the above general formula (1) as an active ingredient, which is used together with a suitable nontoxic pharmaceutical carrier to prepare a general pharmaceutical preparation composition. Formed and put into practical use.

The above-mentioned pharmaceutical carriers used in the pharmaceutical preparation of the present invention include fillers, extenders, binders, humectants, disintegrants, surfactants, and the like, which are usually used depending on the use form of the preparation.
Diluents or excipients such as lubricants can be exemplified, and these are appropriately selected and used depending on the dosage unit form of the obtained preparation.

As the dosage unit form of the pharmaceutical preparation, various forms can be selected according to the purpose of treatment, and typical examples are tablets, pills, powders, solutions, suspensions, emulsions, granules, Capsules, suppositories, injections (solutions, suspensions, etc.),
Ointments and the like.

When formed into tablets, the above-mentioned pharmaceutical carriers include excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, potassium phosphate and the like.
Water, ethanol, propanol, single syrup, glucose solution, starch solution, gelatin solution, binders such as carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone, sodium carboxymethylcellulose, calcium carboxymethylcellulose, low-substituted hydroxypropylcellulose, dried Disintegrants such as starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, surfactants such as polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, and monoglyceride stearate; sucrose, stearin, cocoa butter; Disintegration inhibitors such as hydrogenated oil, quaternary ammonium bases, absorption enhancers such as sodium lauryl sulfate, glycerin, starch and the like Moisturizers, starch, lactose, kaolin, bentonite, adsorbent such as colloidal silicic acid, purified talc, stearates, boric acid powder, a lubricant such as polyethylene glycol can be used. Further, the tablets can be made into tablets coated with a usual coating, if necessary, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets or double tablets or multilayer tablets.

In the case of molding into a pill form, as a pharmaceutical carrier, for example, glucose, lactose, starch, cocoa butter,
Excipients such as hardened vegetable oil, kaolin and talc, gum arabic powder, tragacanth powder, binders such as gelatin and ethanol, and disintegrants such as laminaran and agar can be used.

For shaping in the form of suppositories, for example, polyethylene glycol, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides and the like can be used as pharmaceutical carriers.

Capsules are usually prepared by mixing the active ingredient compound of the present invention with the above-mentioned various pharmaceutical carriers and filling the mixture into hard gelatin capsules, soft capsules and the like according to a conventional method.

When prepared as an injection, such as a liquid, emulsion, or suspension, it is preferable that the preparation be sterilized and isotonic with blood. Water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like can be used. In this case, a sufficient amount of salt, glucose or glycerin to adjust the isotonic solution may be contained in the drug of the present invention, and ordinary solubilizers, buffers, soothing agents and the like are added. May be.

Further, the drug of the present invention may contain a coloring agent, a preservative, a fragrance, a flavoring agent, a sweetening agent and the like and other pharmaceuticals, if necessary.

When shaping into the form of an ointment such as a paste, cream or gel, for example, white vaseline, paraffin, glycerin, a cellulose derivative, polyethylene glycol, silicone, bentonite and the like can be used as a diluent.

The amount of the active ingredient compound represented by the general formula (1) to be contained in the drug of the present invention is not particularly limited and may be appropriately selected from a wide range. It is preferred that the content be contained to a certain degree.

The administration method of the above pharmaceutical preparation is not particularly limited, and is determined according to various preparation forms, age, sex and other conditions of the patient, degree of disease, and the like. For example, tablets, pills, solutions, suspensions, emulsions, granules, and capsules are orally administered, and injections are administered intravenously, alone or as a mixture with ordinary replenishers such as glucose and amino acids. Intramuscularly, intradermally, subcutaneously or intraperitoneally, and suppositories may be administered rectally alone.

The dose of the above pharmaceutical preparation is appropriately selected depending on the usage, age, sex and other conditions of the patient, the degree of the disease and the like.
The dosage is preferably about 0.5 to 20 mg per kg of body weight per day, and the preparation can be administered in 1 to 4 times a day.

[0063]

EXAMPLES In order to explain the present invention in more detail, preparation examples of the nitric oxide synthase inhibitor of the present invention will be given as preparation examples, and then pharmacological test examples.

[0064]

Formulation Example 1 Preparation of Capsule As an active ingredient compound, 5-n-butyl-7- (3,4,
5-trimethoxybenzoylamino) pyrazolo [1,5
-A] Using pyrimidine, 250 per capsule
Hard gelatin capsule containing 1000 mg (1000)
Was prepared according to the following formula.

Active ingredient compound 250 g Microcrystalline cellulose (Japanese Pharmacopoeia product) 30 g Corn starch (Japan Pharmacopoeia product) 17 g Talc (Japanese Pharmacopoeia product) 2 g Magnesium stearate (Japanese Pharmacopoeia product) 1 g Was finely powdered, mixed to form a uniform mixture, and filled in a gelatin capsule having desired dimensions for oral administration to prepare a desired capsule.

[0066]

Formulation Example 2 Preparation of tablet 5-n-butyl-7- (3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] as an active ingredient
Tablets (2000 tablets) containing 300 mg per tablet were prepared using pyrimidine according to the following formula.

Active ingredient compound 600 g Lactose (Japanese Pharmacopoeia) 67 g Corn starch (Japanese Pharmacopoeia) 33 g Carboxymethylcellulose calcium (Japanese Pharmacopoeia) 25 g Methylcellulose (Japanese Pharmacopoeia) 12 g Magnesium stearate (Japanese Pharmacopoeia) 3g That is, according to the above formula, the active ingredient compound, lactose, corn starch and carboxymethylcellulose calcium were sufficiently mixed, the mixture was granulated with an aqueous methylcellulose solution, passed through a 24-mesh sieve, and mixed with magnesium stearate. The tablets were pressed to obtain the target tablets.

[0068]

Formulation Example 3 Preparation of Capsules Using 5-n-butyl-7- (2-trifluoromethylbenzoylamino) pyrazolo [1,5-a] pyrimidine as an active ingredient, 200 mg per capsule is contained. Hard gelatin capsules (2000) were prepared according to the following formulation.

Active ingredient compound 400 g Microcrystalline cellulose (Japanese Pharmacopoeia) 60 g Corn starch (Japanese Pharmacopoeia) 34 g Talc (Japanese Pharmacopoeia) 4 g Magnesium stearate (Japanese Pharmacopoeia) 2 g Was finely powdered, mixed to form a uniform mixture, and filled into gelatin capsules for oral administration having desired dimensions to obtain the desired capsule.

[0070]

Formulation Example 4 Preparation of Tablet Using 5-n-butyl-7- (2-trifluoromethylbenzoylamino) pyrazolo [1,5-a] pyrimidine as an active ingredient, a tablet containing 300 mg per tablet. (2000 tablets) was prepared according to the following formulation.

Active ingredient compound 600 g Lactose (Japanese Pharmacopoeia) 67 g Corn starch (Japanese Pharmacopoeia) 33 g Carboxymethylcellulose calcium (Japanese Pharmacopoeia) 25 g Methylcellulose (Japanese Pharmacopoeia) 12 g Magnesium stearate (Japanese Pharmacopoeia) 3g That is, according to the above formula, the active ingredient compound, lactose, corn starch and carboxymethylcellulose calcium were sufficiently mixed, the mixture was granulated with an aqueous methylcellulose solution, passed through a 24-mesh sieve, and mixed with magnesium stearate. The tablets were pressed to obtain the target tablets.

[0072]

Pharmacological Test Example 1 Male Wistar rats (8 weeks old, 200-250 g) were sacrificed by cervical dislocation, and the thoracic aorta was immediately removed and the surrounding connective tissue was peeled off. Next, in order to eliminate the influence of cNOS existing in the vascular endothelial cells, the endothelial cells were removed by rubbing the vascular lumen with a cotton thread, and cut into 2 mm lengths to obtain ring-shaped specimens. This sample was used in Krebs-Henseleit solution (NaCl
118 mM, KCl 4.7 mM, CaCl ₂ 2.5 m
M, KH ₂ PO ₄ 1.2 mM, MgSO ₄ 1.2 mM,
NaHCO ₃ 25 mM, glucose 11 mM)
l under a pressure of 1 g in the organ bath containing O ₂ / CO ₂
(95% / 5%) The mixed gas was continuously ventilated.

First, with respect to this specimen, 3 × 10 ⁻⁷ M of phenylephrine was added to contract the blood vessel, and then 10 ⁻⁵ M of acetylcholine was added, and 10 ⁻⁵ M of L-arginine was added.
No relaxation occurs in any of the cases where
It was confirmed that both NOS and iNOS were lacking.

Next, 300 ng / ml of lipopolysaccharide (LPS) was added to the above sample, and 8 hours later, 10 ^-5 M of L-arginine was added to measure the relaxation of blood vessels and the cyclic GMP (cGMP) concentration ( Control group).

On the other hand, 30 minutes before the addition of LPS, 5-
3 × 10 ⁻⁵ M of n-butyl-7- (3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine (the compound of Example 1 in Table 1) was placed in an organ bath. Vascular relaxation and cG after addition of L-arginine
The MP concentration was measured in the same manner as described above (the present invention group).

The contraction and relaxation of the blood vessels were measured with an isotonic transducer (TD-111T, manufactured by Nihon Kohden), and a recorder (NIHO) was used.
N DENSI KAGAKU, U-228). Also, cGMP
The concentration was determined using a commercially available radioimmunoassay kit (Amersha
m, cGMP [ ¹²⁵ I] assay system).

As a result, in the group of the present invention, the relaxation of blood vessels was suppressed by 70% as compared with the control group. In addition, the cGMP concentration was also suppressed by 80% in the present invention group as compared with the control group.

From the above, the active ingredient compound of the present invention
It is clear that LPS inhibits the induction of iNOS.

[0079]

Pharmacological Test Example 2 Male Wistar rats (8 weeks old, 200-250 g) were sacrificed by cervical dislocation, and the thoracic aorta was immediately excised and the surrounding connective tissue was peeled off. Blood vessels were treated with collagenase 238 U / ml, esterase 22.5 U / ml, bovine serum albumin 0.2%
HEPES-Hanks solution containing CaCl ₂ .H ₂ O
185.5 mg / l, KCl 400.0 mg / l, K
H ₂ PO ₄ 60.0 mg / l, MgSO ₄ 97.7 mg
/ L, NaCl 8000.0 mg / l, NaHCO ₃ 3
50.0 mg / l, Na ₂ HPO ₄ 47.5 mg / l,
(Glucose 1000 mg / l) at 37 ° C. for 45 minutes.

Next, the endothelial cells and outer membrane were detached from the blood vessel in the HEPES / Hank's solution to remove only the medial smooth muscle, cut into small pieces, and contained the same collagenase, esterase and bovine serum albumin as described above. HEPE
The mixture was incubated in S. Hanks' solution at 37 ° C. for 70 minutes to perform enzyme digestion. This, Dulbecco's modified Eagle's medium containing fetal bovine serum 10% (MgSO ₄ · 7H ₂
O 200.0 mg / l, NaCl 6400 mg / l,
NaHCO ₃ 3700.0 mg / l, NaH ₂ PO ₄ 1
25.5 mg / l, Fe (NO ₃ ) ₃ .9H ₂ O 0.1 m
g / l, phenol red 15.0mg / l, folic acid
4.0 mg / l, nicotinamide 4.0 mg / l, calcium pantothenate 4.0 mg / l, pyridoxal HCl 4.0 mg / l, riboflavin 0.4 mg
/ L, Thiamine.HCl 4.0mg / l, choline chloride
4.0 mg / l, glucose 1000 mg / l, 1-
Inositol 7.0 mg / l, sodium pyruvate
110.0 mg / l), washed several times, 2 × 1
They were inoculated in a Petri dish at a rate of 0 ⁵ / ml. When the cells became confluent, they were passaged and used for the next experiment at the fourth passage.

That is, L-arginine 10 was added to the above cultured cells.
^-5 M was added, and LPS 300 ng / ml or interleukin-1β (IL-1β) 10 ng / ml was further added and left for 24 hours, and the amount of accumulated NO ₂ was measured (control group).

On the other hand, 30 minutes before the addition of LPS or IL-1β, 5-n-butyl-7- (3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine (first In the table, 3 × 10 ⁻⁵ M or 5-n-butyl-7- (2-trifluoromethylbenzoylamino) pyrazolo of the compound of Example 1 of the present invention group 1) [1,5-
a] 3 × 10 ⁻⁵ M of pyrimidine (the compound of Example 32 in Table 1, present invention group 2) was added to an organ bath, and L-
The amount of accumulated NO ₂ 24 hours after the addition of arginine or IL-1β was measured in the same manner as described above (the present invention group).

The amount of NO ₂ was determined by adding an equal amount of Gr to the culture supernatant.
iss reagent (0.1% N- (1-naphthyl) ethylenediamine dihydrochloride / H ₂ O + 1% sulfanylamine / 2.5% H ₃ PO ₄ ) was added, and 570 nm was added.
Quantitation was performed by measuring the absorbance at

The results are shown in FIG. 1 and FIG.

FIG. 1 shows data when LPS was added, and FIG. 2 shows data when IL-1β was added. In each figure, data of a group to which nothing was added as a control is also shown.

From FIGS. 1 and 2, it is apparent that the active ingredient compound of the present invention inhibits iNOS induction by LPS and IL-1β.

[0087]

[Pharmacological Test Example 3] Spraque Dawl
ey) Male rats (6-9 weeks old, 200-250 g) were sacrificed by cervical dislocation, and the thoracic aorta was immediately excised and the surrounding connective tissue was detached. Next, this was divided into 5 to 7 slices, each of which was cut vertically, and then the endothelial cells were removed by rubbing the vascular lumen with a cleaning swab in order to eliminate the influence of cNOS present in the vascular endothelial cells. Then, a specimen was prepared.

A Krebs-Henseleit solution (pharmacological test example 1) was prepared by adding a dimethyl sulfoxide solution of the active ingredient compound (test compound) of the present invention prepared at a concentration of 30 μM, and further adding L-arginine to a concentration of 400 μM. (The same composition as the sample), and incubated at 37 ° C. for 30 minutes. Subsequently, lipopolysaccharide (LPS) was added at a concentration of 1000 ng / ml and incubated at 37 ° C. for 24 hours (experimental group using test compounds, present invention group).

Next, the supernatant was placed on a 96-well plate and described in the literature [Shinogen Chemistry Experiment Course 10, vascular, endothelial and smooth muscle, 135
Page, the Japanese Biochemical Society, Tokyo Kagaku Dojin, in accordance with NO ₂ measurement method described in 1993], the NO ₂ were developed with grease (Griess) solution, Bio-Kinetic Sleater (Biokinet
ics reader, BIO-TEK Instruments, EL-340
And the amount of accumulated NO ₂ was calculated.

Further, a blood vessel piece of the specimen was dissolved in a 1N aqueous solution of sodium hydroxide, and a Bio-Rad DC protein assay kit (Bio-Rad DC protein assay kit,
d Laboratories) and color spectrophotometer (Spectrophotometer, HITACHI, U-300)
0 type) to calculate the amount of protein. From these values, the amount of NO ₂ produced per mg of protein was determined.

On the other hand, the same test was repeated for a control group to which dimethyl sulfoxide was added instead of the test compound and a control group to which no LPS was added.

From the NO ₂ production amount per mg of protein in each group obtained as described above, the iNOS induction inhibition rate was calculated according to the following equation.

Inhibition rate (%) = ｛1-[(value of the present invention group) −
(Control group value)] / [(Control group value)-(Control group value)]｝ × 100 The obtained results are shown in Table 6.

According to Table 6, the active ingredient compound of the present invention
It is clear that it inhibits iNOS induction by PS.

[0095]

[Table 1]

[0096]

[Table 2]

[0097]

[Table 3]

[0098]

[Table 4]

[0099]

[Table 5]

[0100]

[Table 6]

[0101]

[Table 7]

[0102]

[Table 8]

[0103]

[Table 9]

[0104]

[Table 10]

[0105]

[Table 11]

[0106]

[Table 12]

[0107]

[Table 13]

[0108]

[Table 14]

[0109]

[Table 15]

[0110]

[Table 16]

[0111]

[Table 17]

[Brief description of the drawings]

FIG. 1 is a graph showing the inhibitory effect of the active ingredient compound of the present invention on LPS-induced iNOS induction determined according to Pharmacological Test Example 2.

FIG. 2 is a graph showing the inhibitory effect of the active ingredient compound of the present invention on iNOS induction by IL-1β determined according to Pharmacological Test Example 2.

Claims (9)

[Claims] 1. A compound of the general formula Wherein R ¹ is a hydrogen atom, a thienyl group, a lower alkoxy group, a lower alkylthio group, a oxo group or a lower alkyl group which may have a hydroxyl group as a substituent, a cycloalkyl group, a thienyl group, a furyl group, a lower alkenyl. R ² represents a phenyl group which may have 1 to 3 groups selected from a lower alkyl group, a lower alkoxy group, a phenylthio group and a halogen atom as a group or a substituent; R ² represents a naphthyl group, a cycloalkyl group, a furyl group, a thienyl group; Group, a pyridyl group that may be substituted with a halogen atom, a phenoxy group that may be substituted with a halogen atom or a lower alkyl group, a lower alkoxy group, a halogen atom, a nitro group, a halogen-substituted lower alkyl group, Substituted lower alkoxy group, lower alkoxycarbonyl group,
A phenyl group which may have 1 to 3 of a group selected from a hydroxyl group, a phenyl lower alkoxy group, an amino group, a cyano group, a lower alkanoyloxy group, a phenyl group and a di-lower alkoxyphosphoryl lower alkyl group,
R ³ represents a hydrogen atom, a phenyl group or a lower alkyl group;
⁴ is a hydrogen atom, a lower alkyl group, a lower alkoxycarbonyl group, a phenyl lower alkyl group, a phenyl group or a halogen atom which may have a phenylthio group as a substituent, R ⁵ is a hydrogen atom or a lower alkyl group, R ⁶ is A hydrogen atom, a lower alkyl group, a phenyl lower alkyl group or a benzoyl group having 1 to 3 groups selected from a lower alkoxy group, a halogen-substituted lower alkyl group and a halogen atom as a substituent, and R ¹ and R ⁵ are It may combine with each other to form a lower alkylene group, Q represents a carbonyl group or a sulfonyl group, A represents a single bond, a lower alkylene group or a lower alkenylene group, and n represents 0 or 1. A pyrazolo [1,5-a] pyrimidine derivative represented by the formula (I) as an active ingredient, and an effective amount thereof together with a nontoxic carrier. 2. The compound according to claim 1, wherein R ⁵ and R ⁶ are each a hydrogen atom, Q is a carbonyl group, A is a single bond, and n is 0. The nitric oxide synthase inhibitor according to claim 1. 3. The active ingredient according to claim 1, wherein R ¹ is a phenyl group or a lower alkyl group which may have a hydroxyl group or a lower alkoxy group as a substituent, and R ² is a substituent. a lower alkoxy group, a phenyl group having 1 to 3 groups selected from a halogen-substituted lower alkyl group and a halogen atom, nitric oxide synthesis according to claim 2 R ⁴ is a compound which is a hydrogen atom or a phenyl group Enzyme inhibitors. 4. The active ingredient according to claim 1, wherein R ¹ is a phenyl group, methyl group, ethyl group, n-butyl group or n-pentyl group, and R ² is 4-ethoxy-3. ,
The monoxide according to claim 2, wherein the compound is a 5-dimethoxyphenyl group, 3,4,5-trimethoxyphenyl group, 2-methoxyphenyl group, 2,4-dichlorophenyl group or 2-trifluoromethylphenyl group. Nitrogen synthase inhibitors. 5. The method according to claim 5, wherein the active ingredient is 5-n-butyl-7-.
(3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine, 5-phenyl-7-
(3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine and 5-n-butyl-7-
The nitric oxide synthase inhibitor according to claim 4, which is a compound selected from (2-trifluoromethylbenzoylamino) pyrazolo [1,5-a] pyrimidine. 6. The active ingredient is 5-n-butyl-7-
The nitric oxide synthase inhibitor according to claim 5, which is (3,4,5-trimethoxybenzoylamino) pyrazolo [1,5-a] pyrimidine. 7. The nitric oxide synthase inhibitor according to claim 1, which selectively inhibits inducible nitric oxide synthase. 8. The pyrazolo [1,5-
a) A preventive and therapeutic agent for sepsis, which comprises an effective amount of a pyrimidine derivative together with a non-toxic carrier. 9. The pyrazolo [1,5-
a) An endotoxin shock ameliorating agent comprising an effective amount of a pyrimidine derivative together with a non-toxic carrier.