JPH0667838B2 - Lipid peroxide production inhibitor - Google Patents

Description

TECHNICAL FIELD The present invention relates to a lipid peroxide production inhibitor, and more specifically, to prevention of various ischemic diseases and various brain diseases, heart diseases and peripheral circulatory disorders associated therewith. The present invention relates to a lipid peroxide production inhibitor useful as a therapeutic agent.

[Prior Art and Problems Thereof] In circulatory disorder disease in the brain, heart or periphery, arachidonic acid and other unsaturated fatty acids released from the cell membrane due to ischemia (a state where blood is not supplied to the tissue) Active oxygen generated in the surrounding tissues (OH ^● radical superoxide, etc.) acts to produce lipid peroxide. Such changes progress not only during ischemia but also at an accelerated rate by blood-mediated reoxygenation after ischemia resumption, resulting in injury of living cells rich in unsaturated fatty acids, destruction of surrounding tissues, and destruction of vascular endothelium. , Causing vasospasm or edema,
It is known that the vicious cycle of these series of reactions progresses the pathology ("Cerebral ischemia and cell injury" edited by Takao Asano, edited by Neuron, 1980; "Brain ischemia and free radicals" edited by Takao Asano, edited by Neuron. Company, 1983).

Therefore, if the production of lipid peroxide due to active oxygen is suppressed,
It is possible to prevent tissue destruction, vascular endothelium destruction, vasospasm, edema, etc., and is a new type of circulation that acts on the cause of disease, completely different from conventional drugs that improve circulation by increasing blood flow. It becomes a preventive and / or therapeutic agent for disorders. In recent years, in particular, the effectiveness of increasing blood flow in the infarct region has been questioned, and it is said that it is rather counterproductive in acute-stage cerebrovascular accidents, and such drugs have become more important.

As a drug that suppresses lipid peroxidation due to active oxygen, vitamin E, the following formula: Idebenone (Biochemical and Biophysical Research Communications (Bioc
hmeical and Biophysical Research Communication
s) 125, 1046 (1984); Takeda Research Institute 44, 30 (1985)) and the following formula: In shown is Nizofenon (Journal of Neuro-Chemistry (Journal of Neurochemistry) 37, 934
(1981)) is known.

However, vitamin E has an insufficient action, idebenone and nizophenone have a long synthetic pathway, and idebenone is difficult to be solubilized in water, which may cause a problem in injectable preparation. Nizophenone has an inhibitory effect on the central nervous system. Is strong (Pharmaceutical Research 16 , 1 (1985)).

Various pyrazolone derivatives are known.

JP-A-51-13766 discloses the following formula (A): And a use thereof as an antithrombotic agent; JP-A-59-141517 discloses a compound for treating infarction, inflammation, asthma, etc. after myocardial ischemia. The use is; in Japanese Patent Laid-Open No. 59-175469, the following formula: (In the formula, X represents a group —CH ₂ CH ₂ O—, —CH ₂ CH ₂ S—,
R represents an aryl group. ) Is used as a pyrazolin-5-one derivative and its use as a lipoxygenase inhibitor; JP-B-59-512 discloses the following formula: (In the formula, R ₁ represents a hydrogen atom or an amino group, R ₂ represents an aryl group, and X represents a group —CH ₂ CH ₂ O—, etc.), and a pyrazolin-5-one derivative thereof and a diuretic thereof. Agent,
It is used as an antihypertensive agent and an antithrombotic agent; in German Patent Application No. 2836891, the following formula: (In the formula, R ₁ and R ₂ represent a hydrogen atom or a substituent.) And a use as a pyrazolin-5-one derivative and an anti-inflammatory agent; JP-A-52-31083 discloses formula: (In the formula, R ₁ and R ₂ represent a hydrogen atom or a substituent, and R ₃
Represents a lower alkyl group. ), Its use as an analgesic and anti-inflammatory agent, and the following formula as a starting material thereof: (In the formula, R ₁ and R ₂ have the same meanings as described above.) A compound represented by: is disclosed in JP-A-52-268. (In the formula, R ₁ and R ₂ represent a hydrogen atom or a substituent.) And its use as a therapeutic agent for buoyancy, hypertension and thrombosis, and a diuretic agent; JP-A-42-19593 In the official gazette, the following formula: The following formula of the compound represented by: Although the use of the anti-inflammatory analgesic represented by the formula (1) as a synthetic intermediate is described, there is no description about the action of suppressing lipid peroxidation by active oxygen. Further, the compound represented by the formula (A) is effective in a model of ischemic heart disease using rats, rabbits and dogs, but is ineffective in a model using pigs which is close to the circulatory dynamics of human heart, the results are well consistent with the report that it is invalid to ischemic heart disease in humans (European journal of Pharma roller diisopropyl (European journal of Pharmacology) 114, 1
89 (1985)). Further, there is no description of a specific action regarding protection after resumption of cerebral ischemia.

Therefore, as a result of intensive studies conducted by the present inventors for the purpose of providing a drug having an action of suppressing lipid peroxidation caused by active oxygen, the following formula (I): (In the formula, R ₁ represents a hydrogen atom, an aryl group, an alkyl group having 1 to 5 carbon atoms or an alkoxycarbonylalkyl group having 3 to 6 total oxygen; R ₂ represents a hydrogen atom, an aryloxy group, an arylmercapto. A group, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 1 to 3 carbon atoms; or, R ₁ and R ₂ jointly represent an alkylene group having 3 to 5 carbon atoms; and R ₃ is Heterocycles which are unsubstituted or substituted with 1 to 3 substituents which are the same or different and are selected from the group consisting of alkyl groups having 1 to 5 carbon atoms, alkoxy groups having 1 to 5 carbon atoms, aryl groups and halogen atoms. The pyrazolone derivative represented by () represents a strong inhibitory effect on peroxidation, and has a protective effect such as recovery of EEG in an animal model of cerebral ischemia resumption that is close to the actual pathological condition. Complete the present invention This has led to the.

[Structure of the Invention] The lipid peroxide production inhibitor of the present invention is characterized by comprising a pyrazolone derivative represented by the above formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound (I) used in the present invention has the following formula (I ′) or (I ″): It can also have the structure shown in. Therefore, the above formula (I ′)
Alternatively, the compound having the structure of (I ″) is also included in the active ingredient of the present invention.

In formula (I), examples of the aryl group in the definition of R ₁ include a phenyl group and a phenyl group substituted with a substituent such as a methyl group, a butyl group, a methoxy group, a butoxy group, a chlorine atom and a hydroxyl group. . The alkyl group having 1 to 5 carbon atoms in the definition of R ₁ and R ₂ is a methyl group,
Examples thereof include ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, tert-butyl group, pentyl group and the like. Examples of the alkoxycarbonylalkyl group having 3 to 6 carbon atoms in the definition of R ₁ include a methoxycarbonylmethyl group, an ethoxycarbonylmethyl group, a propoxycarbonylmethyl group, a methoxycarbonylethyl group and a methoxycarbonylpropyl group. R ₂
As the aryloxy group in the definition of, phenoxy group, p-methylphenoxy group, p-methoxyphenoxy group,
Examples thereof include p-chlorophenoxy group and p-hydroxyphenoxy group, and examples of the arylmercapto group include phenylmercapto group, p-methylphenylmercapto group, p-methoxyphenylmercapto group, p-chlorophenylmercapto group and p-hydroxyphenyl group. Examples thereof include a mercapto group. Examples of the hydroxyalkyl group having 1 to 3 carbon atoms in the definition of R ₂ include a hydroxymethyl group, a 2-hydroxyethyl group and a 3-hydroxypropyl group. The heterocyclic residue in the definition of R ₃ is preferably a monocyclic heterocyclic residue having 5 to 6 members containing heteroatoms 1 to 2 and a condensed heterocyclic residue containing the monocyclic heterocycle. Such monocyclic heterocyclic residue, a furyl group, a thienyl group, a pyrrolyl group, a pyrrolinyl group, a pyrrolidinyl group, an oxazolyl group, an isoxazolyl group, a thiazolyl group, an isothiazolyl group, an imidazolyl group, an imidazolinyl group, an imidazolidinyl group, a pyrazolyl group, 5 such as pyrazolinyl group and pyrazolidinyl group
And a 6-membered ring residue such as a pyridyl group, a pyrimidinyl group, a pyrazinyl group, a pyranyl group, a piperidyl group, a pyridazinyl group, a piperazinyl group and a morpholinyl group. The condensed heterocyclic residue is preferably a condensed heterocyclic residue in which the monocyclic heterocycle is condensed with a benzene ring, for example, a quinolyl group, an isoquinolyl group, a phthalazinyl group or a benzothiazolyl group. These heterocyclic residues are substituted with the same or different 1 to 3 substituents selected from the group consisting of an alkyl group having 1 to 5 carbon atoms, an alkoxy group having 1 to 5 carbon atoms, an aryl group and a halogen atom. May be. Examples of the alkyl group having 1 to 5 carbon atoms in the substituent of the heterocyclic residue include methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group,
tert-butyl group, and examples of the alkoxy group having 1 to 5 carbon atoms include a methoxy group, an ethoxy group, a propoxy group,
Examples thereof include an isopropoxy group and a butoxy group, and examples of the aryl group include a phenyl group, a methyl group, a methoxy group,
Examples thereof include a phenyl group substituted with a substituent such as a halogen atom, and examples of the halogen atom include a fluorine atom, a chlorine atom, and a bromine atom.

Specific examples of the compound (I) used in the present invention include the compounds shown below.

3-Methyl-1- (2-pyridyl) -2-pyrazolin-5-one 3,4-dimethyl-1- (2-pyridyl) -2-pyrazolin-5-
On 3-propyl-1- (2-pyridyl) -2-pyrazolin-5-one 3-phenyl-1- (2-pyridyl) -2-pyrazolin-5-one 3-methyl-4-phenoxy-1- ( 2-Pyridyl) -2-pyrazolin-5-one 3-methyl-1- (6-methyl-2-pyridyl) -2-pyrazolin-5-one 1- (6-butyl-2-pyridyl) -3-methyl -2
-Pyrazolin-5-one 1- (6-methoxy-2-pyridyl) -3-methyl-
2-Pyrazolin-5-one 1- (6-butoxy-2-pyridyl) -3-methyl-
2-Pyrazolin-5-one 1- (3-chloro-2-pyridyl) -3-methyl-2-pyrazolin-5-one 1- (5-chloro-2-pyridyl) -3-methyl-2-pyrazolin- 5-one 1- (6-chloro-2-pyridyl) -3-methyl-2-pyrazolin-5-one 3-methyl-1- (4-pyridyl) -2-pyrazolin-5-one 3,4-dimethyl -1- (4-pyridyl) -2-pyrazolin-5-
On 3-Methyl-1- (2-pyrazinyl) -2-pyrazolin-5-one 3-phenyl-1- (2-pyrazinyl) -2-pyrazolin-5-
On 3-methyl-1- (2-pyrimidinyl) -2-pyrazolin-5-
On 3-phenyl-1- (2-pyrimidinyl) -2-pyrazolin-
5-one 3-methyl-1- (3-pyridazinyl) -2-pyrazolin-5-
On 3-methyl-1- (6-methyl-3-pyridazinyl) -2-pyrazolin-5-one 1- (6-methoxy-3-pyridazinyl) -3-methyl-2-pyrazolin-5-one 1- ( 6-Butoxy-3-pyridazinyl) -3-methyl-2-pyrazolin-5-one 3-methyl-1- (6-phenyl-3-pyridazinyl) -2-pyrazolin-5-one 1- (6-methoxy- 3-pyridazinyl) -3-phenyl-2-
Pyrazolin-5-one 1- (6-methoxy-3-pyridazinyl) -3-methyl-2-pyrazolin-5-one 1- (6-chloro-5-methyl-3-pyridazinyl) -3-methyl-2- Pyrazolin-5-one 3-Methyl-1- (6-phenyl-3-pyridazinyl) -2-pyrazolin-5-one 1- (2-Imidazonilyl) -3-methyl-2-pyrazolin-
5-one 1- (2-imidazolylyl) -3-phenyl-2-pyrazolin-5-one 3-methyl-1- (2-thiazolyl) -2-pyrazolin-5-one 3-methyl-1- (4- Methyl-2-thiazolyl) -2-pyrazolin-5-one 1- (4-butyl-2-thiazolyl) -3-methyl-
2-Pyrazolin-5-one 3-methyl-1- (4-phenyl-2-thiazolyl) -2-pyrazolin-5-one 1- (4-methyl-2-thiazolyl) -3-phenyl-2-pyrazolin- 5-one 3-methyl-1- (2-quinolyl) -2-pyrazolin-5-one 3-methyl-1- (4-quinolyl) -2-pyrazolin-5-one 3-methyl-1- (4- Methyl-2-quinolyl) -2-pyrazolin-5-one 1- (7-chloro-4-quinolyl) -2-pyrazolin-5-one 1- (7-chloro-4-quinolyl) -3-methyl-2 -Pyrazolin-5-one 3-phenyl-1- (2-quinolyl) -2-pyrazolin-5-one 1- (1-isoquinolyl) -3-methyl-2-pyrazolin-5-
On 3-methyl-1- (1-phthalazinyl) -2-pyrazolin-5-
On 3-Methyl-1- (4-methyl-1-phthalazinyl) -2-pyrazolin-5-one 3-Methyl-1- (4-phenyl-1-phthalazinyl) -2-pyrazolin-5-one 3-phenyl -1- (1-phthalazinyl) -2-pyrazoline-
5-one 1- (2-benzothiazolyl) -3-methyl-2-pyrazolin-5-one 1- (2-benzothiazolyl) -3,4-dimethyl-2-pyrazolin-5-one 1- (2-benzothiazolyl) -3-Phenyl-2-pyrazolin-5-one Of the salts of compound (I) used in the present invention, pharmaceutically acceptable salts include mineral acids such as hydrochloric acid, sulfuric acid, hydrobromic acid and phosphoric acid. Salts of methanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, acetic acid, glycolic acid, glucuronic acid, maleic acid, fumaric acid, oxalic acid, ascorbic acid, citric acid, salicylic acid, nicotinic acid, tartaric acid, etc. Salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as magnesium and calcium; ammonia, tris (hydroxymethyl) aminomethane, N, N-bis (hydroxyethyl) piperazi 2-amino-2-methyl-1-propanol, ethanolamine, N- methylglucamine, and salts with amines such as L- glucamine.

The compound used in the present invention can be synthesized by any purposeful method, but one preferable method is shown below.

(In the formula, R ₁ , R ₂ and R ₃ have the same meanings as described above, and R ′ represents an alkyl group having 1 to 5 carbon atoms.) That is, the β-keto acid derivative represented by the formula (II) and the formula (II
The hydrazine derivative represented by I) is required in the presence of a solvent such as alcohols such as methanol and ethanol or aromatics such as benzene and toluene, or without a solvent. If necessary, potassium carbonate, sodium ethoxide, etc. , Sodium hydroxide, potassium hydroxide, sodium acetate and other bases, hydrochloric acid, sulfuric acid, hydrogen bromide and other mineral acids, acetic acid,
In the presence of a catalyst such as an organic acid such as paratoluenesulfonic acid,
Compound (I) can be obtained by reacting at a temperature of 10 to 200 ° C.

When clinically applying Compound (I), when used orally, 1 to 100 mg of Compound (I) once for an adult
It is preferable to administer 1 to 3 times a day. In the case of intravenous injection, 0.01 to 10 mg of Compound (I) is administered once to an adult.
It is preferable to administer 2 to 5 times a day or continuous infusion of these doses. In the case of rectal administration, 1 to 100 mg of Compound (I) is administered 1 to 3 times a day. preferable. Further, it is more preferable that the above dose is appropriately increased or decreased depending on age, disease state, and symptom.

In the case of oral or rectal administration, it may be used as a sustained release preparation.

Upon formulation, one or more compounds (I) or pharmaceutically acceptable salts thereof are usually used as a composition containing a commonly used pharmaceutical carrier, excipient or other additive. is there. The pharmaceutical carrier may be solid or liquid,
Examples of the solid carrier include lactose, white clay (kaolin), sucrose, crystalline cellulose, corn starch, talc, agar, pectin, acacia, stearic acid, magnesium stearate, lecithin, sodium chloride and the like.

Examples of liquid carriers include syrup, glycerin, peanut oil, polyvinylpyrrolidone, olive oil, ethanol, benzyl alcohol, propylene glycol, water and the like.

It can take various dosage forms, and when a solid carrier is used, it can be tablets, powders, granules, hard gelatin capsules, suppositories or troches. The amount of solid carrier may vary widely but preferably will be from about 1 mg to about 1 g.

When a liquid carrier is used, it can be a syrup, emulsion, soft gelatin capsule, sterile injectable solution such as ampoules, or aqueous or non-aqueous suspension.

In addition, the compound (I) can also be used by performing an operation such as putting it in a cyclodextrin inclusion complex or a liposome.

[Effect of the Invention] The lipid peroxide production inhibitor of the present invention has an excellent action,
Various ischemic diseases or various diseases based on them, that is,
Cerebral infarctions such as cerebral infarction and stroke, or various cerebral diseases such as cerebral dysfunction caused by them, vascular dementia, cerebral vascular tissue lesions with aging, myocardial infarction, various heart diseases based on myocardial ischemia such as heart failure It is also useful as a prophylactic / therapeutic agent for various peripheral circulatory disorders.

[Examples of the Invention] Hereinafter, the present invention will be described in more detail based on Synthesis Examples and Examples, but these do not limit the scope of the present invention.

Synthesis Example 1 Synthesis of 3-methyl-1- (2-pyridyl) -2-pyrazolin-5-one 5.96 g of ethyl acetoacetate was added to 5 g of 2-hydrazinopyridine, and heated and stirred at 100 ° C. for 2 hours. After cooling, it was recrystallized from ethanol to obtain 5.44 g of 3-methyl-1- (2-pyridyl) -2-pyrazolin-5-one (Compound No. 1) as colorless crystals.

Yield 68% Melting point 111-112 ° C Synthesis Examples 2-21 In the same manner as in Synthesis Example 1, compounds shown in Table 1 as Compound Nos. 2-21 were synthesized.

Example 1 Inhibition of Lipid Peroxidation (a) Preparation of Brain Homogenate Using Wistar male rats, a brain homogenate was prepared according to the following procedure. The chest was opened under anesthesia by intraperitoneal administration of pentobarbital sodium 45 mg / Kg, and a polyethylene tube was inserted into the aorta from the left atrium and fixed. It was then ice-cooled through this tube
Cerebral perfusion was performed with 50 mM phosphate buffered saline (pH 7.4) (hereinafter referred to as "PBS"), and the whole brain was removed. After removing the cerebellum, measure the wet weight of the cerebrum, add 9 times the amount of PBS,
It was homogenized by crushing with a Teflon homogenizer in ice water. This brain lonegenate at 2200rpm at 4 ℃
After centrifuging for 10 minutes, 0.3 ml of the supernatant was collected in a light-shielding test tube with a stopper, and brain honegenate for drug evaluation.

(B) Evaluation of test drug To the brain homogenate prepared in (a), 0.6 ml of PBS and 10 μl of a test drug in a tanol solution (final concentration 500 μM or 0.3 to 100)
μM concentration 3) was added and heated in a 37 ° C. warm bath for 30 minutes. Then, after adding 200 μl of 35% perchloric acid aqueous solution, centrifugation was performed at 2600 rpm for 10 minutes at 4 ° C. to obtain a supernatant. Also, for blank measurement, 10 μl of ethanol was added in place of 10 μl of the ethanol solution of the test drug (blank), and the same operation was performed.

(C) Quantification of lipid peroxide To 0.1 ml of the supernatant obtained in (b), 0.2 ml of 8.1% sodium dodecyl sulfate aqueous solution, 20 ml of 20% acetate buffer (pH 3.5), 0.67
% 2-Thiobarbituric acid aqueous solution (1.5 ml) and distilled water (0.7 ml) were added and mixed. Then, this mixture was heated in a boiling water bath for 60 minutes, rapidly cooled with ice water, 1.0 ml of distilled water and 5.0 ml of a pyridine-butanol mixture (1:15) were added, and after shaking for about 30 seconds, at 3000 rpm for 10 minutes. After centrifugation, the supernatant was used as a lipid peroxide measurement sample. In addition, lipoperoxide-
Test (Lipoperoxide-test) (Wako Pure Chemical Industries, Ltd .; 1,1,
0.1 ml of 3,3-tetraethoxypropane (containing 5 nmol / ml) was added in place of the brain homogenate obtained in (b) to prepare a standard solution.

Lipid peroxide is a fluorescence spectrophotometer (produced by Hitachi, Ltd. 204
Type), and measuring at an excitation wavelength of 515 nm and a fluorescence wavelength of 550 nm,
The amount of lipid peroxide (TBA value) was calculated according to the following formula.

F: Fluorescence intensity of standard solution f: Fluorescence intensity of test drug Then, the inhibition rate of each concentration of the test drug against the TBA value of the blank in (b) was determined, and the IC ₅₀ value was calculated according to the least squares method. The results are shown in Table 1.

Example 2 Formulation of Lipid Peroxide Production Inhibitor of the Present Invention (1) Tablet The following ingredients were mixed according to a conventional method and tableted using a conventional device.

Active ingredient of the present invention (Compound No. 1) 10 mg Crystalline cellulose 21 mg Corn starch 33 mg Lactose 65 mg Magnesium stearate 1.3 mg (2) Soft capsule The following ingredients were mixed according to a conventional method and filled into a soft capsule.

Active ingredient of the present invention (Compound No. 1) 10 mg Olive oil 105 mg Lecithin 6.5 mg (3) Injectable preparation The following ingredients were mixed according to a conventional method to prepare a 1 ml ampoule.

Active ingredient of the present invention (Compound No. 1) 0.7 mg Sodium chloride 3.5 mg Distilled water for injection 1.0 ml

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location A61K 31/50 7431-4C 31/505 7431-4C // C07D 401/04 231 7602-4C 403 / 04 231 7602-4C 417/04 231 9051-4C (C07D 401/04 213: 00 8217-4C 231: 00) 7431-4C (C07D 401/04 217: 00 7431-4C 231: 00) 7431-4C (C07D 401/04 215: 00 7019-4C 231: 00) 7431-4C (C07D 403/04 231: 00 7431-4C 237: 00) 8615-4C (C07D 403/04 231: 00 7431-4C 239: 00) 8615 -4C (C07D 403/04 231: 00 7431-4C 241: 00) 8615-4C (C07D 403/04 231: 00 7431-4C 233: 00) 9360-4C (C07D 417/04 231: 00 7431-4C 277 : 00) 9051-4C (72) Inventor Toshishi Sachi 8-5-1 Chuo, Ami-cho, Inashiki-gun, Ibaraki Mitsubishi Oil Kayaku Co., Ltd. Research Laboratory (72) Inventor Yoko Sakurai 8-5-1 Chuo, Ami-machi, Inashiki-gun, Ibaraki Prefecture Mitsubishi Petrochemical Co., Ltd. Research Laboratory

Claims (1)

[Claims] 1. The following formula: (In the formula, R ₁ represents a hydrogen atom, an aryl group, an alkyl group having 1 to 5 carbon atoms or an alkoxycarbonylalkyl group having 3 to 6 carbon atoms; R ₂ represents a hydrogen atom, an aryloxy group, an arylmercapto. A group, an alkyl group having 1 to 5 carbon atoms or a hydroxyalkyl group having 1 to 3 carbon atoms; or, R ₁ and R ₂ jointly represent an alkylene group having 3 to 5 carbon atoms; and R ₃ is Heterocycles which are unsubstituted or substituted with 1 to 3 substituents which are the same or different and are selected from the group consisting of alkyl groups having 1 to 5 carbon atoms, alkoxy groups having 1 to 5 carbon atoms, aryl groups and halogen atoms. A pyrazolone derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof is used as an active ingredient.