JPH09227377A - Scavenger of active oxygen and free radical - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a scavenger of active oxygen or free radical having a hydroxy radical removing effect and useful for treating a disease, etc., associated with active oxygen or free radical. SOLUTION: This scavenger contains a hydantoin derivative of the formula (R1 and R2 are each H, an alkyl or a cycloalkyl; X and Y are each H, an alkyl, OH or an alkoxy, or X and Y together form an oxo group) or its salt as an active component. As an example of a compound of the formula, 5- hydroxy-1-methylhydantoin is cited. The scavenger is effective for the treatment of various kinds of diseases such as myocardial infarction, reperfusion damage, cerebral infarction, autoimmune disease, fibrosis, pulmonary disease, skin disease. arthritis, side effect of an anticancer agent, radiation hazard, bacterial shock, inflammatory disease and cataract. The scavenger has little side effect and can be easily formulated. The daily dose of the scavenger for adult is 1-1000mg, preferably 5-600mg of the active component in oral administration. The dose in the case of parenteral administration, e.g., in the case of an injection, is sufficiently from a third to a tenth of the dose in oral administration.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel active oxygen / free radical scavenger containing a hydantoin derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0002]

2. Description of the Related Art In recent years, there has been strong interest in damage to biomolecules, membranes, tissues caused by free radicals and active oxygen, and various diseases derived therefrom. Free radicals are chemical species that have unpaired electrons and are unstable and highly reactive. Examples of active oxygen species include superoxide, hydroxy radical, hydrogen peroxide, and singlet oxygen. Among them, superoxide and hydroxy radical are radical species, and hydroxy radical is known as a highly reactive free radical. Has been.
In vivo target molecules for damage due to free radicals and active oxygen are diverse, including lipids, nucleic acids, enzymes, proteins, and sugars. In particular, highly unsaturated fatty acids localized in the lipids of cell membranes are susceptible to attack and oxidative modification. As a result, lipid peroxide is produced. As a result, the cell membrane is damaged, and not only the destruction of the membrane structure but also the enzymatic action of the protein localized and active there and the receptor function are greatly damaged. It is also known that the produced lipid peroxide is locally released into the blood and causes secondary lesions such as vascular lesions.

Many reports have been made on the relationship between free radicals and active oxygen and diseases, and the usefulness of substances that eliminate free radicals and active oxygen as pharmaceuticals has received a great deal of attention. In vivo superoxide scavenger SOD
(Superoxide dismutase) is a typical free radical / active oxygen scavenging substance, and its development as a drug is in progress. The therapeutic targets of SOD include myocardial infarction, reperfusion injury, cerebral infarction, autoimmune disease (collagen disease, Behcet's disease, ulcerative colitis), fibrosis, lung disease (pulmonary edema, pulmonary fibrosis), skin disease ( Various diseases such as burns, trauma, keloids, sun sensitivity, dermatitis), joint diseases, side effects of anticancer agents, radiation damage, bacterial shock, inflammatory diseases, and cataracts are mentioned (for example, “pathology of free radicals”). Biochemistry and Clinics, Inflammation and Anti-Inflammation ", Japanese Clin, 46, No. 10, pp. 93-97, 1988).

[0004]

Since SOD is a protein preparation and has problems such as a short half-life in blood and difficulty in being taken up into cells, various preparation techniques have been studied. The current situation. Therefore, it has been desired to develop a new free radical / active oxygen scavenger that is easy to formulate and has few side effects.

[0005]

The object of the present invention is to provide such a novel free radical / active oxygen scavenger.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION The active ingredient of the active oxygen / free radical scavenger of the present invention is a hydantoin derivative represented by the following general formula (I).

Embedded image [In the formula, R ₁ and R ₂ are the same or different and each represents hydrogen, an alkyl group or a cycloalkyl group, and X and Y are the same or different, each is hydrogen, an alkyl group, a hydroxyl group or an alkoxy group, or X and Y. Represents an oxo group. ]

In the above general formula (I), R ₁ and R ₂
Each is the same or different and is hydrogen, an alkyl group, preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, neopentyl, t-pentyl, hexyl, dimethylbutyl, heptyl, Octyl, nonyl,
A linear or branched alkyl group having 1 to 20 carbon atoms such as decyl or stearyl, or a cycloalkyl group, preferably having 3 to 8 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. Represents a cycloalkyl group.

X and Y are the same or different and each is hydrogen, an alkyl group, preferably a linear or branched carbon atom having 1 to 3 carbon atoms such as methyl, ethyl, propyl and isopropyl.
A linear or branched carbon number 1 such as an alkyl group, a hydroxyl group or an alkoxy group, preferably methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, t-butoxy, pentoxy, isopentoxy, neopentoxy, etc. To alkoxy groups, or X and Y represent an oxo group.

Preferred compounds in the present invention are as follows. [Compound 1] Hydantoin [Compound 2] 1-Methylhydantoin [Compound 3] 3-Methylhydantoin [Compound 4] 1-Ethylhydantoin [Compound 5] 1-Propylhydantoin [Compound 6] 1-Butylhydantoin [Compound 7] 1 -T-Butylhydantoin [Compound 8] 1-hexylhydantoin [Compound 9] 1- (1,3-dimethylbutyl) hydantoin [Compound 10] 1-decylhydantoin [Compound 11] 1-stearylhydantoin [Compound 12] 1, 3-Dimethylhydantoin [Compound 13] 1,5-Dimethylhydantoin [Compound 14] 3,5-Dimethylhydantoin

[Compound 15] 1-Cyclopentylhydantoin [Compound 16] 1-Cyclohexylhydantoin [Compound 17] 1-Cyclohexyl-3-methylhydantoin [Compound 18] 3-Cyclohexylhydantoin [Compound 19] 1,3-Dicyclohexylhydantoin

[Compound 20] 5-hydroxyhydantoin [Compound 21] 5-hydroxy-1-methylhydantoin [Compound 22] 5-hydroxy-3-methylhydantoin [Compound 23] 5-hydroxy-1-ethylhydantoin [Compound 24] ] 5-Hydroxy-1-propylhydantoin [Compound 25] 5-Hydroxy-1-butylhydantoin [Compound 26] 5-Hydroxy-1-t-butylhydantoin [Compound 27] 5-Hydroxy-1-hexylhydantoin [Compound 28] ] 5-Hydroxy-1- (1,3-dimethylbutyl) hydantoin [Compound 29] 5-Hydroxy-1-decylhydantoin [Compound 30] 5-Hydroxy-1-stearylhydantoin [Compound 31] 5-Hydroxy-1- Cyclopentylhydan In [Compound 32] 5-hydroxy-1-cyclohexyl hydantoin [Compound 33] 5-hydroxy-1-cyclohexyl -
3-Methylhydantoin [Compound 34] 5-Hydroxy-1,3-dimethylhydantoin [Compound 35] 5-Hydroxy-1,5-dimethylhydantoin [Compound 36] 5-Hydroxy-3,5-dimethylhydantoin [Compound 37] 5-hydroxy-1,3-dicyclohexylhydantoin

[Compound 38] 5-methoxyhydantoin [Compound 39] 5-methoxy-1-methylhydantoin [Compound 40] 5-methoxy-3-methylhydantoin [Compound 41] 5-methoxy-1-ethylhydantoin [Compound 42] 5-methoxy-1-propylhydantoin [Compound 43] 5-methoxy-1-butylhydantoin [Compound 44] 5-methoxy-1-cyclohexylhydantoin [Compound 45] 5-methoxy-3-cyclohexylhydantoin [Compound 46] 5 -Ethoxyhydantoin [Compound 47] 5-Ethoxy-1-methylhydantoin [Compound 48] 5-Ethoxy-3-methylhydantoin [Compound 49] 5-Ethoxy-1-ethylhydantoin [Compound 50] 5-Ethoxy-1-propyl Hydantoin [Compound 51] 5-Ethoxy-1-butylhydantoin [Compound 52] 5-Propoxyhydantoin [Compound 53] 5-Propoxy-1-methylhydantoin [Compound 54] 5-Propoxy-3-methylhydantoin [Compound 55] 5- Propoxy-1-ethylhydantoin [Compound 56] 5-Propoxy-1-propylhydantoin [Compound 57] 5-Propoxy-1-butylhydantoin [Compound 58] 5-Butoxyhydantoin [Compound 59] 5-Butoxy-1-methylhydantoin [Compound 60] 5-Butoxy-3-methylhydantoin [Compound 61] 5-t-butoxyhydantoin [Compound 62] 5-t-butoxy-1-methylhydantoin [Compound 63] 5-t-butoxy-3-butylhydantoin

[Compound 64] Imidazolidinetrione [Compound 65] 1-Methylimidazolidinetrione [Compound 66] 1-Ethylimidazolidinetrione [Compound 67] 1-Butylimidazolidinetrione [Compound 68] 1-Isobutylimidazolidinetrione [Compound 69] 1-t-butylimidazolidinetrione [Compound 70] 1-hexylimidazolidinetrione [Compound 71] 1- (1,3-dimethylbutyl) imidazolidinetrione [Compound 72] 1-decylimidazolidinetrione [Compound 72] Compound 73] 1-Cyclopentylimidazolidinetrione [Compound 74] 1-Cyclopentyl-3-ethylimidazolidinetrione [Compound 75] 1-Cyclohexylimidazolidinetrione [Compound 76] 1,3-Dimethylimidazo Jintorion [Compound 77] 1,3-dicyclohexyl-imidazolidine-trione

The hydantoin derivative of the present invention includes the salt represented by the above general formula (I), for example, hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, phosphoric acid, perchloric acid, thiocyanic acid, boric acid, Formic acid, acetic acid, haloacetic acid, propionic acid, glycolic acid, citric acid, tartaric acid, succinic acid, gluconic acid, lactic acid, malonic acid, fumaric acid, anthranilic acid, benzoic acid,
Addition salts with acids such as cinnamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid and sulfanilic acid, alkali metals such as sodium and potassium, alkaline earth metals such as calcium, magnesium and barium, or aluminum and zinc. And salts with metals and the like. These salts can be prepared from the free hydantoin derivative of the present invention by a known method, or can be converted into each other.

The present invention is also applicable when stereoisomers such as cis-trans isomers, optical isomers and conformational isomers exist in the compounds of the present invention, or when they exist in the form of hydrates or complex compounds. Any of its stereoisomers, hydrates,
It also includes complex compounds. The hydantoin derivative of the present invention and the method for producing the same are disclosed in JP-A-61-212275, JP-A-62-14 and the like.

The compound of the present invention can be made into a medicament by appropriately combining with a suitable pharmaceutical carrier or diluent.
It can be formulated by any ordinary method, and can be formulated as an oral preparation such as tablets, capsules, powders and liquids, or as a parenteral preparation for subcutaneous, intravenous, intramuscular, rectal and intranasal administration. it can. In the formulation, the compound of the present invention may be used in the form of a pharmaceutically acceptable salt thereof, the compound of the present invention can be used alone or in an appropriate combination, and a compounding agent with other pharmaceutically active ingredient can be used. May be

For oral administration preparations, as they are or together with suitable additives, for example, conventional excipients such as lactose, mannitol, corn starch, potato starch, calcium citrate and the like, cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose and the like. , Binders such as gum arabic, corn starch and gelatin, disintegrants such as corn starch, potato starch, carboxymethyl cellulose calcium, lubricants such as talc and magnesium stearate, other extenders, wetting agents, buffers, preservatives Tablets, powders, granules or capsules can be prepared by appropriately combining agents and flavors.

As the injection, an aqueous solvent such as distilled water for injection, physiological saline, or glucose injection, or a solution, suspension or non-aqueous solvent such as vegetable oil, synthetic fatty acid glyceride, higher fatty acid ester or propylene glycol is used. It can be an emulsion, and if necessary, commonly used additives such as a solubilizing agent, a tonicity agent, a suspending agent, an emulsifying agent, a stabilizer and a preservative may be appropriately added. Furthermore, depending on the type of disease and the patient, other dosage forms, syrups and
It can be formulated into suppositories, inhalants, aerosols, eye drops, external preparations (ointments, gels, patches, etc.) and the like.

Although the desirable dose of the compound of the present invention varies depending on the administration subject, dosage form, administration method, administration period and the like, in order to obtain a desired effect, generally, the dose of the active ingredient is 1 to 1 per day for an adult. 1000 mg, preferably 5 to 600 m
It can be administered orally. In the case of parenteral administration (eg, injection), generally, a smaller amount can be expected to be effective than oral administration, and therefore, for example, 3 to 1/10 of the above oral dose is expected.
Dose levels of 10% are considered sufficient.

Formulation examples of pharmaceutical compositions containing the compound of the present invention as an active ingredient are shown below, but the present invention is not limited thereto.

[Table 1]

Preferred embodiments of the active oxygen / free radical scavenger of the present invention containing the compound represented by the general formula (I) as an active ingredient will be described below. (1) An active oxygen / free radical scavenger containing, as an active ingredient, a compound in which one of X and Y in the general formula (I) is hydrogen. (2) An active oxygen / free radical scavenger in which, of the compounds described in (1) above, the other one of X and Y is a hydroxyl group. (3) An active oxygen / free radical scavenger in which one of R ₁ and R ₂ is an alkyl group and the other is hydrogen among the compounds described in (2) above. (4) An active oxygen / free radical scavenger in which R ₁ is an alkyl group among the compounds described in (3) above. (5) Of the compounds described in (4) above, R ₁ has ₁ carbon atom.
An active oxygen / free radical scavenger having 4 to 4 alkyl groups. (6) An active oxygen / free radical scavenger of the compound described in (5) above, wherein R ₁ is a methyl group.

(7) An active oxygen / free radical scavenger containing, as an active ingredient, a compound of the general formula (I) in which both X and Y are hydrogen. (8) Among the compounds described in (7) above, an active oxygen / free radical scavenger in which one of R ₁ and R ₂ is an alkyl group and the other is hydrogen. (9) An active oxygen / free radical scavenger in which R ₁ is an alkyl group among the compounds described in (8) above. (10) An active oxygen / free radical scavenger in which R ₁ is an alkyl group having 1 to 4 carbon atoms, among the compounds described in (9) above. (11) An active oxygen / free radical scavenger of the compound described in (10) above, wherein R ₁ is a methyl group.

(12) Of the compounds described in (1) above,
An active oxygen / free radical scavenger in which the other one of X and Y is an alkoxy group. (13) An active oxygen / free radical scavenger in which R ₁ is an alkyl group and R ₂ is hydrogen among the compounds according to the above (12). (14) An active oxygen / free radical scavenger of the compound described in (13) above, wherein R ₁ is a methyl group.

(15) Among the compounds described in (1) above,
An active oxygen / free radical scavenger in which the other one of X and Y is an alkyl group. (16) An active oxygen / free radical scavenger in which R ₁ is an alkyl group and R ₂ is hydrogen, among the compounds described in (15) above. (17) An active oxygen / free radical scavenger of the compound described in (16) above, wherein R ₁ is a methyl group.

(18) An active oxygen / free radical scavenger containing, as an active ingredient, a compound represented by the general formula (I), wherein X and Y are oxo groups. (19) An active oxygen / free radical scavenger in which R ₁ is an alkyl group and R ₂ is hydrogen among the compounds described in (18). (20) An active oxygen / free radical scavenger of the compound described in (19) above, wherein R ₁ is a methyl group. (21) A hydroxy radical scavenger containing a compound represented by the general formula (I) as an active ingredient. (22) A hydroxy radical scavenger containing the compound described in each of (1) to (20) above as an active ingredient.

A particularly preferred compound is 5-hydroxy-1-methylhydantoin, which has been confirmed to have low toxicity and almost no side effects.

[0027]

【Example】

(1) Measurement of hydroxy radical scavenging ability The hydroxy radical scavenging ability of the test substance was measured by using the ESR spin trapping method which is commonly used for the detection of hydroxy radicals. The generation of hydroxy radicals
The Fenton reaction in which divalent iron chelated with DTPA (diethyleneamine-pentaacetic acid) reacts with hydrogen peroxide to generate hydroxy radicals was performed. DM
PO (dimethyl-pyrroline-N-oxide) reacts rapidly with the generated hydroxyl radical, and the product of the reaction shows a four-line ESR signal with a characteristic intensity ratio of 1: 2: 2: 1. When a test substance having a hydroxy radical scavenging action coexists in this reaction system, the above-mentioned characteristic signal is suppressed, and thus the hydroxy radical scavenging ability can be measured using this as an index.

That is, 75 μl of a mixed solution of 1 mM ferrous sulfate and DTPA [dissolved in 100 mM phosphate buffer (pH 7.8)] was placed in a test tube, and 50 μl of a sample solution containing a test substance. , And 1 mM or 2 mM DMP
20 μl of O was added, and finally 1 mM hydrogen peroxide solution 75
The reaction was initiated by the addition of μl. The hydrogen peroxide solution was prepared immediately before each experiment and the reaction was performed at room temperature. The ESR spectrum in this reaction system was measured by a conventional method. DMP
The concentration of the test substance (IC ₅₀ ) at which the signal intensity by O-OH was suppressed to 50% in the presence of the test substance as compared with the case of the absence was determined.

An example of the results obtained with 1 mM DMPO is shown below. Dimethyl sulfoxide, which is known to have a hydroxy radical scavenging action, is shown as a comparative drug.

[Table 2]

(2) Inhibitory action of endothelial cells by activated leukocytes The inhibitory action of cell damage by hydroxy radicals was measured using the following experimental system. The vascular endothelial cells obtained by enzymatic treatment from bovine descending aorta were cultured in a 48-well plate using MEM Earle containing 15% fetal bovine serum. To the confluent endothelial cells, ⁵¹ Cr (sodium chromate) 3.7 KBq was added and cultured for 18 hours to incorporate ⁵¹ Cr into the endothelial cells. After removing the chromium in the medium by washing, the test drug, human leukocytes and phorbol ester (phorbol myristate acetate) were added, and the mixture was cultured for 6 hours. The radioactivity of chromium released into the medium from endothelial cells damaged by activated leukocytes was measured by a γ counter, and the damage rate of endothelial cells was calculated by the following formula.

The failure rate in the absence of the test drug is 1
An example of the result when it is set to 00% (control group) is shown in FIG.

FIG.

[0032]

From the above test results, it was shown that the substance of the present invention has a hydroxy radical scavenging action. In the pharmacological test (1), when the concentration of DMPO added was changed, the IC _{50 of the} compound of the present invention was also changed in proportion to the change. Therefore, the hydroxy radical scavenging action of the present compound is a competitive antagonistic action with DMPO. It was suggested that there is.

Therefore, the hydantoin derivative of the present invention having an active oxygen / free radical scavenging action is a disease associated with active oxygen or free radicals, such as myocardial infarction, reperfusion injury, cerebral infarction, which is mentioned as an indication of SOD. Autoimmune diseases (collagen disease, Behcet's disease, ulcerative colitis), fibrosis, lung diseases (pulmonary edema, pulmonary fibrosis), skin diseases (burns, trauma, keloids, sun sensitivity, dermatitis), arthritis, anticancer Side effects of the drug, radiation damage, bacterial shock,
It is useful as a drug for treating various diseases such as inflammatory diseases and cataracts. The compound of the present invention, unlike dimethyl sulfoxide, which has a cytotoxic effect, has extremely low toxicity, is highly safe, and can be orally administered, so that it can be used for chronic diseases requiring long-term administration. Is highly useful.

[Brief description of drawings]

FIG. 1 shows an example of the results showing the inhibitory effect of the compound of the present invention on the cell damage caused by hydroxy radical.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication location A61K 31/415 ABJ A61K 31/415 ABJ ABL ABL ABS ABS ACD ACD ADA ADA AD ADU ADU ADU ADZ // C07D 233/74 C07D 233/74 233/80 233/80 233/96 233/96

Claims (1)

[Reference number] PC-261 [Claims] 1. An active oxygen / free radical scavenger containing a hydantoin derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. Embedded image [In the formula, R ₁ and R ₂ are the same or different and each represents hydrogen, an alkyl group or a cycloalkyl group, and X and Y are the same or different, each is hydrogen, an alkyl group, a hydroxyl group or an alkoxy group, or X and Y. Represents an oxo group. ]