JPH0447646B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to antioxidants. For living organisms, oxygen is essential for maintaining life, such as energy production and metabolism. The oxygen becomes so-called active oxygen species such as oxygen anion radicals, peroxide ions, and hydroxyl radicals through reactions in energy production systems, oxygen reactions, reactions with ultraviolet rays, radiation, and the like. While these reactive oxygen species are useful for living organisms, such as oxygenating enzymes and bactericidal effects on white blood cells, they also contain unsaturated substances that form phospholipids in biological membranes, such as oleic acid, linoleic acid, linolenic acid, and arachidonic acid, which are abundant in living organisms. Promotes peroxidation of fatty acids and forms lipid peroxide. This lipid peroxide causes generation of alkoxy radicals and hydroxyl radicals like the above-mentioned active oxygen species, attacks biological membranes, and causes membrane damage and deactivation of various useful enzymes [Metabolism, 15 (10)] ,
(See 1978 Special Feature on Active Oxygen). However, there are enzymes involved in the metabolic deactivation of the above-mentioned reactive oxygen species, such as superoxide dismutase (SOD), catalase, and glutathione peroxidase, and enzymes such as α-tocopherol (vitamin E), etc. There are various vitamins and other substances that have antioxidant abilities, and their effects normally maintain the body's normal body, but for some reason, the enzymes, vitamins, etc. mentioned above may cause a deficiency in the appropriate defense mechanism. It is often observed that the generation of reactive oxygen species and the production and accumulation of lipid peroxides occur in a manner that exceeds the capacity of these defense mechanisms. When a deficiency in such a defense mechanism occurs, the peroxidation reaction progresses in a chain reaction, leading to serious disorders such as various diseases caused by platelet aggregation, inflammation, liver damage, arteriosclerosis, hemolysis, aging and senile diseases, retinopathy,
Lung damage, heart and lung damage caused by certain drugs, ischemic vascular disease, etc. occur. It scavenges active oxygen species (radicals), which have traditionally been thought to be the main cause of the various disorders listed above.
Compounds that have the effect of preventing or reducing the production and accumulation of lipid peroxides in living bodies are generally called antioxidants, and there have been many reports of their use in preventing and treating the various diseases listed above.
Reported antioxidants include enzyme agents such as the above-mentioned SOD [Superoxide and Medicine, Yoshihiko Oyanagi, 1981, Kyoritsu Shuppansha, pp. 137-141], butylated hydroxytoluene (BHT), and butylated hydroxyl. Anisole (BHA), α-tocopherol (vitamin E), etc. [Makoto Mino, Hidetaka Tanaka, Pharmaceutical Journal, 19 (12), 1983, p. 2351-2359 and Toshihiko Suematsu, Ibid., 19 (5), 1983, p909-914〕
There is. The present inventors have discovered that a series of indane derivatives newly produced by the present inventors have an antioxidant effect similar to that of the above-mentioned conventionally known antioxidants, and have hereby completed the present invention. I came to the conclusion. That is, the present invention is based on the general formula [In the formula, R ¹ represents an amino group, and R ² and R ³ are the same or different and represent a hydrogen atom, a lower alkyl group, or a halogen atom, respectively. ] The present invention relates to an antioxidant containing an indane derivative represented by the following and its salt as an active ingredient. The antioxidant active ingredient compound of the present invention has the effect of removing active oxygen species and preventing or reducing lipid peroxide production in the body. Therefore, the anti-acid agent of the present invention can be used as a prophylactic and therapeutic agent for various disorders and diseases caused by excessive generation of active oxygen species, accumulation of lipid peroxides in the body, or lack of defense mechanism against these, for example, anti-acid Pharmaceuticals such as sclerosing agents, cancer prevention agents, anticancer agents, anti-inflammatory agents, analgesics, autoimmune disease treatment agents, platelet aggregation inhibitors, antihypertensive agents, antihyperlipidemia agents, and agents for preventing and treating retinopathy of prematurity and cataracts. It is useful as Furthermore, the antioxidant of the present invention can be used not only as the above-mentioned pharmaceuticals, but also as
For example, it is effective for use as an antioxidant for fats and oils contained in processed foods. The above general formula (1) used as an active ingredient of the antioxidant of the present invention
In the indane derivative represented by R ₂ and
The lower alkyl group represented by R ₃ includes methyl,
ethyl, propyl, isopropyl, butyl, tert
-butyl, pentyl, hexyl, etc. with 1 or more carbon atoms
Examples include straight chain or branched alkyl groups of 6.
Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. The indane derivative represented by the above general formula (1) is a new compound that has not been described in any literature, and can be produced, for example, by the method shown in the following reaction scheme-1. [In the formula, R ² and R ³ are the same as above. ] Known compound of general formula (2) and hydroxyamine (3)
The reaction with can be carried out in a suitable inert solvent in the presence or absence of a basic compound. Basic compounds used in this case include, for example, inorganic basic compounds such as sodium hydroxide, potassium hydroxide, sodium carbon, potassium carbon, etc., piperidine, pyridine, triethylamine, 1,5-diazabicyclo[4,3,0] Nonen-5 (DBN),
1,5-diazabicyclo[5,4,0]undecene (DBU), 1,4-diazabicyclo[2,2,
2] Examples include organic bases such as octane (DABCO). Any inert solvent may be used as long as it does not adversely affect the reaction, but examples include lower alcohols such as methanol, ethanol, and isopropanol, and ethers such as dioxane, tetrahydrofuran, diethyl ether, and ethylene glycol monomethyl ether. Examples include aromatic hydrocarbons such as benzene, toluene, and xylene, hydrogen halides such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride, dimethylformamide, dimethylsulfoxide, and hexamethylphosphoric triamide. The amount of hydroxylamine (3) to be used is usually at least equimolar, preferably equimolar to 5 times the molar amount of the compound of general formula (2). The reaction temperature is
The temperature is usually room temperature to 200°C, preferably 50 to 150°C, and the reaction is generally completed in about 1 to 10 hours. Reduction of the compound of general formula (1a) can be carried out by catalytic hydrogenation in a suitable solvent in the presence of a catalyst. The solvent used is
For example, water, alcohols such as acetic acid, methanol, ethanol, and isopropanol, hydrocarbons such as hexane and cyclohexane, ethers such as diethylene glycol dimethyl ether, dioxane, tetrahydrofuran, and diethyl ether, esters such as ethyl acetate and methyl acetate, and dimethylformamide. Examples include aprotic polar solvents such as. Further, as the catalyst, for example, palladium, palladium-black, palladium-carbon, platinum, platinum oxide, copper chromite, Raney nickel, etc. are used. The amount of the catalyst to be used is generally about 0.02 to 1 times the amount of the compound of general formula (1a). The reaction temperature is usually -20°C to around room temperature, preferably 0°C to around room temperature, the hydrogen pressure is usually 1 to 10 atm, and the reaction is generally completed in about 0.5 to 10 hours. In addition, the indane derivative represented by the general formula (1) is
It can also be produced by the method shown in Reaction Scheme-2 below. [In the formula, R ¹ is the same as above. R ² ' and R ³ ' are the same or different and represent a hydrogen atom or a lower alkyl group. however
R ² ′ and R ³ ′ cannot both be lower alkyl groups. R ² ″ and R ³ are the same or different and represent a hydrogen atom, a lower alkyl group, or a halogen atom. However, R ² ″ and R ³ ″ cannot both be a lower alkyl group,
In addition, at least one of R ² ″ and R ³ ″ represents a halogen atom. ] The halogenation reaction of the compound of general formula (1c) is carried out in the presence of a conventional halogenating agent. A wide variety of known halogenating agents can be used as the halogenating agent used in such a reaction, such as halogen molecules such as bromine and chlorine, iodine monochloride, sulfuryl chloride, thionyl chloride, N-bromosuccinimide, N-chlorosuccinimide, etc. Examples include halogenating agents such as N-halogenosuccinimide and the like. The amount of the halogenating agent to be used is usually about 1 molar to 10 times, preferably 1 molar to 5 times the molar amount of the compound of general formula (1c). Examples of the solvent used in the reaction include dichloromethane, dichloroethane, chloroform, halogenated hydrocarbons such as carbon tetrachloride, acetic acid, propionic acid, water, and the like. In this reaction, the reaction temperature is usually 0°C to the boiling point of the reaction solvent,
Preferably the temperature is 0 to 40°C, usually 1 to 10°C.
The reaction completes in about an hour. The target products obtained in each step can be easily isolated and purified by conventional separation means. Examples of the separation means include solvent extraction, dilution, recrystallization, column chromatography, and preparative thin layer chromatography. Note that the compound of general formula (1) naturally includes optical isomers. The indane derivative represented by general formula (1) can be easily converted into an acid addition salt by the action of a pharmaceutically acceptable acid, and in the present invention, this acid addition salt can also be used as an active ingredient. can. In the above, examples of acids include hydrochloric acid, sulfuric acid, phosphoric acid,
Inorganic acids such as hydrobromic acid, organic acids such as acetic acid, oxalic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, malonic acid, methanesulfonic acid, benzoic acid, etc. can be used. In the present invention, the antioxidant is usually used in the form of a common pharmaceutical preparation. The formulation is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and lubricants. Various forms of this pharmaceutical preparation can be selected depending on the therapeutic purpose, and representative examples include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, and injections ( (solutions, suspensions, etc.), ointments, etc. When forming tablets, a wide variety of carriers known in this field can be used, including excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, and silicic acid. agent, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, binder such as polyvinylpyrrolidone, dried starch, sodium alginate, agar powder, laminaran powder, Sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch,
Disintegrants such as lactose, disintegration inhibitors such as sucrose, stearin, cocoa butter, hydrogenated oil, etc., absorption enhancers such as quaternary ammonium bases and sodium lauryl sulfate, humectants such as glycerin and starch, starch, lactose, kaolin Examples include adsorbents such as bentonite and colloidal silicic acid, purified talc, stearate, boric acid powder, and lubricants such as polyethylene glycol. Furthermore, the tablets may be coated with a conventional coating, if necessary, such as sugar-coated tablets, gelatin-encapsulated tablets, enteric-coated tablets, film-coated tablets, double tablets, or multilayer tablets. When shaping into a pill, a wide variety of conventionally known carriers can be used, such as excipients such as glucose, lactose, starch, cacao butter, hydrogenated vegetable oil, kaolin, talc, powdered gum arabic, powdered tragacanth,
Examples include binders such as gelatin and ethanol, and disintegrants such as laminar agar. When forming into a suppository, a wide variety of conventionally known carriers can be used, such as polyethylene glycol, cacao butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, and the like. When prepared as injections, solutions and suspensions are preferably sterilized and isotonic with blood, and when forming these solutions, emulsions, and suspensions, this agent is used as a diluent. All those commonly used in the field can be used, including water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and the like. In this case, a sufficient amount of salt, glucose, or glycerin to prepare an isotonic solution may be included in the pharmaceutical preparation, and usual solubilizing agents, buffers, and soothing sugars may be added. It's okay. Furthermore, coloring agents, preservatives, perfumes, flavoring agents, sweeteners, etc., and other pharmaceuticals may be included in the pharmaceutical preparation, if necessary. When forming pastes, creams and gels, a wide variety of diluents conventionally known in this field can be used, such as white petrolatum, paraffin,
Examples include glycerin, cellulose derivatives, polyethylene glycol, silicon, and bentonite. General formula (1) to be contained in the pharmaceutical formulation of the present invention
The amount of the compound and its salt is not particularly limited and can be appropriately selected from a wide range, but usually 1.
It is preferable that the amount is in the range of ~70% by weight. The method of administering the above pharmaceutical preparation is not particularly limited, and is appropriately determined depending on the various preparation forms, age, sex and other conditions of the patient, the severity of the disease, and the like. For example, tablets, pills, solutions, suspensions, emulsions, granules, and capsules are administered orally. In the case of an injection, it is administered intravenously alone or mixed with a normal replacement fluid such as glucose or amino acids, and furthermore, if necessary, it is administered alone intramuscularly, intradermally, subcutaneously, or intraperitoneally. Suppositories are administered rectally. The dosage of the above pharmaceutical preparation is appropriately selected depending on the usage, patient's age, sex and other conditions, degree of disease, etc., but usually the amount of the compound of general formula (1) and its salt, which is the active ingredient, per day's body weight. Approximately 0.2 to 200 per 1 kg
It is best to use around mg. Below, examples of production of the indane derivative represented by general formula (1) and formulation examples of the antioxidant of the present invention containing the same are listed, followed by pharmacological test examples and toxicity test examples. Production Example 1 Add 28 g of hydroxylamine hydrochloride and 56 g of potassium carbonate to 400 ml of methanol, and heat under reflux for 30 minutes. After cooling, the supernatant is separated and a hydroxylamine methanol solution is prepared. Add 7-hydroxy-4- to this hydroxylamine methanol solution.
Add 16.2 g of methyl-1-indanone and heat under reflux for 5 hours while stirring. The reaction mixture is concentrated to dryness under reduced pressure. Add 200 ml of ethyl acetate to the residue to remove insoluble materials. The liquid was concentrated to dryness under reduced pressure, and the residue was crystallized from methanol to give colorless needle-like crystals of 7-hydroquine-4-methyl-1-indanone oxime.
Obtain 17.6g. mp.148-149.5°C The compound of Production Example 2 is obtained in the same manner as Production Example 1 using appropriate starting materials. Production example 2 4,6-dimethyl-7-hydroxy-1-indanone oxime mp.155-156℃ Colorless needle crystals Production example 3 15.0 g of 7-hydroxy-4-methyl-1-indanone oxime was added to 200 ml of acetic acid. Dissolve, add 1.0 g of platinum oxide catalyst, and heat at room temperature under 5 atmospheres of hydrogen pressure.
Time contact reduction. After removing the catalyst, the liquid is concentrated to dryness under reduced pressure. Add 200 ml of ethanol to the residue, dissolve it, and saturated it by blowing in hydrochloric acid gas. The solvent was concentrated to dryness under reduced pressure, and the residue was recrystallized from ethanol to obtain 3.30 g of 1-amino-7-hydroxy-4-methylindane hydrochloride in the form of colorless needles. mp.221-223°C Compounds listed in Table 1 below are obtained in the same manner as in Production Example 3 using appropriate starting materials.

[Table] Production Example 8 Dissolve 1 g of 1-amino-7-hydroxy-4-methylindane hydrochloride in 20 ml of water, and while stirring vigorously at room temperature - 3N containing 0.85 g of iodine chloride.
Add 5 ml of hydrochloric acid solution dropwise. After stirring at the same temperature for 2 hours, cool on ice. The precipitated crystals are collected and converted into hydrochloride, then washed with ether and dried. 0.70 g of 1-amino-7-hydroxy-6-iodo-4-methylindane hydrochloride is obtained in the form of yellow needles. Example of decomposition production at mp.200℃ or higher 9 1-Amino-7-hydroxyindan hydrochloride
Dissolve 1.0 g in 60 ml of acetic acid, and add 1.53 g of sulfuryl chloride dropwise while stirring under ice cooling. After stirring at the same temperature for 3 hours, the mixture was concentrated under reduced pressure. The residue was dissolved in 50 ml of ethanol saturated with hydrochloric acid gas, and then dried under reduced pressure. Recrystallization from isopropanol-ether gives 0.43 g of colorless prismatic crystals of 1-amino-4,6-dichloro-7-hydroxyindan hydrochloride. mp.238-239℃ (decomposition) Formulation example 1 Compound of Production Example 1 200mg Grape, etc. 250mg Distilled water for injection Appropriate amount Total volume 5ml After dissolving the compound obtained in Production Example 1 and glucose in distilled water for injection, 5ml The mixture is injected into an ampoule, replaced with nitrogen, and sterilized under pressure at 121°C for 15 minutes to obtain an injection having the above composition. Formulation Example 2 Compound of Production Example 3 100g Avicel (trade name, manufactured by Asahi Kasei Corporation) 40g Cornstarch 30g Magnesium stearate 2g TC-5 (trade name, manufactured by Shin-Etsu Chemical Factory Co., Ltd., hydroxypropyl methyl cellulose) 10g Polyethylene glycol - 6000 3g Castor oil 40g Methanol 40g The compound obtained in Production Example 33, Avicel, cornstarch and magnesium stearate are mixed and polished, and then tableted using a sugar coater with a radius of 10mm. the obtained tablets
A film coated tablet having the above composition is produced by coating with a film coating agent consisting of TC-5, polyethylene glycol-6000, castor oil and methanol. Formulation Example 3 Compound of Production Example 7 2g Purified lanolin 5g White beeswax 5g White petrolatum 88g Total amount 100g White beeswax was heated to make it liquid, then the compound obtained in Production Example 7, purified lanolin and white petrolatum were added to make it liquid. After heating until , the mixture is stirred until it begins to solidify to obtain an ointment having the above composition. Pharmacological Tests It is well known that luminol is a luminescent reagent that emits intense light due to hydrogen peroxide in the presence of a hemin catalyst. This test uses linoleic acid hydroperoxide, which is known to be a strong oxidizing agent like hydrogen peroxide, to determine how much the compound to be tested (antioxidant) inhibits the oxidation of luminol by this peroxide. The antioxidative ability of the compound was determined by measuring the amount. Vitamin E (VE), which was used as a control drug (antioxidant), is known to reduce blood peroxidized lipids induced by alloxan in vivo, based on its antioxidant ability. It is recognized that compounds with similar antioxidant ability can similarly reduce blood peroxidized lipids. The test method is as follows. (1) Test method Test compound at a concentration of 1 to 1 x 10 ^-6 mg/ml and linoleic acid hydroperoxide at a concentration of 1.0 x 10 ^-9
Prepare a methanol solution containing a concentration of mol/ml (hereinafter referred to as test solution). Also, 1 x 10 ^-4 M luminol in 0.1 M sodium carbonate buffer, and
FCS (fetal bovine serum, Gibco) at 1.25×10 ^-6
Prepare a 0.1 M sodium carbonate buffer containing a concentration of g/ml. These prepared solutions were inhaled and mixed in the order of test solution, FCS solution, and luminol solution in 0.4 ml portions into an automatic droplet using the flow system shown in Figure 1. After mixing the final luminol solution, a photo counter (R649S : Hamamatsu Photonics Co., Ltd.) to measure the amount of luminescence. In Figure 1, 1 is a photo counter, 2 is a cell, 3 is a mixer, and 4 is a photo counter.
5 is the test solution, 5 is the luminescent reagent (luminol solution)
, 6 is a catalyst (FCS solution), 7 is a washing buffer (0.1M sodium carbonate buffer), 8 is a syringe, 9 is a drain, and 10 is a valve. Each compound shown in Table 2 was used as a test compound at a predetermined concentration, and the results of measuring the amount of luminescence at each concentration are also shown in Table 2. Each luminescence amount was calculated as a percentage (%) with respect to the luminescence amount when the above-mentioned test solution containing no test compound was used as 1 according to the following formula, and the value was shown as the value. Luminescence amount = C-B/A-B x 100 (%) A: Number of counts when no test compound is added and lipid peroxide is added B: Number of counts when neither test compound nor lipid peroxide is added C: Count number when test compound and lipid peroxide are added together

[Table] The test compounds in Table 2 are as follows. The present invention...obtained in Production Example 3 [general formula (1),
R ₁ = NH ₂ , R ₂ = H, R ₃ = CH ₃ ] The present invention...What was obtained in Production Example 4 [General formula (1),
R ₁ = NH ₂ , R ₂ = CH ₃ , R ₃ = CH ₃ ] The present invention...What was obtained in Production Example 6 [General formula (1),
R ₁ = NH ₂ , R ₂ = I, R ₃ = CH ₃ ] The present invention...What was obtained in Production Example 7 [General formula (1),
R ₁ = NH ₂ , R ₂ = Cl, R ₃ = Cl] Control BHT...butylated hydroxytoluene Control VE...vitamin E (2) In addition, in the same test above, linoleic acid hydroperoxide 1 x 10 ^-9 mol/ 50μ in ml
The results of determining the 50% inhibition rate (that is, the concentration of each test compound that inhibits the oxidation ability of the peroxide by 50%, the antioxidant ability IL ₅₀ ) for each test compound are shown in Table 3 below.

[Table] From Tables 2 and 3 above, it can be seen that the indane derivatives used in the present invention (the present invention) all have strong antioxidant ability. Furthermore, each of the above compounds was found to exhibit strong activity in vivo as well as BHT and VE. This shows that the antioxidant of the present invention is useful as a prophylactic and therapeutic agent for various diseases caused by lipid peroxides and reactive oxygen species. 1- obtained by
Amino-7-hydroxy-4-methylindane hydrochloride was administered orally to ICR male mice. As a result, the above compound
The _LD50 value was 600mg/Kg. Almost the same results were obtained for other compounds of the present invention. From this, the compound of the present invention can reduce excessive generation of active oxygen species.
It can be said that there are no serious problems in terms of toxicity at doses that are effective or higher in the treatment of various diseases caused by bioaccumulation of lipid peroxides or deficiencies in defense mechanisms against lipid peroxides.

[Brief explanation of the drawing]

FIG. 1 shows a flow path diagram of an apparatus for measuring the antioxidant ability of the active ingredient compound of the antioxidant of the present invention.

Claims (1)

[Claims] 1. General formula [In the formula, R ¹ represents an amino group, and R ² and R ³ are the same or different and represent a hydrogen atom, a lower alkyl group, or a halogen atom, respectively. ] An antioxidant containing an indane derivative or a salt thereof as an active ingredient.