JP4814501B2 - Antioxidant function improver - Google Patents

Description

  The present invention relates to an anti-oxidant function improving agent useful for preventive treatment of lifestyle diseases such as arteriosclerosis, cancer, diabetes, abnormal pigmentation such as stains and freckles, skin inflammation, and skin aging.

  It is widely known that active oxygen typified by superoxide radical and hydrogen peroxide has cytotoxicity and causes cancer, rheumatism, stains, wrinkles and the like. Further, it is known that LDL carrying cholesterol is changed to oxidized LDL by active oxygen, and the changed oxidized LDL causes arteriosclerosis.

Therefore, it is said that a drug having an antioxidant action is useful as a prophylactic and therapeutic agent for lifestyle-related diseases such as arteriosclerosis, cancer, diabetes, abnormal pigmentation such as stains and freckles, skin inflammation, and skin aging. Various antioxidant components have been found. For example, natural products such as vitamin E and vitamin C, synthetic products such as BHT (3,5-tert-butyl-4-hydroxytoluene) and BHA (2,3-tert-butyl-hydroxyanisole), herbal medicines, etc. It is known to have an antioxidant effect (Patent Document 1).
Japanese Patent Laid-Open No. 10-139678

However, although vitamin E and vitamin C are safe as natural products, their effects as antioxidants are insufficient, and BHT and BHA have problems such as suspected carcinogenicity. Moreover, the effect as an antioxidant was insufficient for those derived from herbal medicines.
Accordingly, an object of the present invention is to provide an antioxidant function improver that can be safely ingested for a long time.

  Thus, the present inventors have studied the pharmacological action of various amino acids, and as a result, superoxide dismutase (SOD) in which highly safe δ-amino acids represented by 5-aminolevulinic acid are known as antioxidant enzymes. Or the glutathione peroxidase (GPx) activity was improved, it discovered that it had the outstanding antioxidant function improvement, and came to complete this invention.

  That is, the present invention provides an antioxidant function improver comprising a δ-amino acid, a derivative thereof or a salt thereof as an active ingredient.

  ADVANTAGE OF THE INVENTION According to this invention, the antioxidant function of the animal in which the antioxidant function declined, for example, the animal including the person in which the antioxidant function fell with aging, can be improved, arteriosclerosis, cancer, diabetes, etc. It is possible to prevent or ameliorate life-style related diseases, stains, freckles and other abnormal pigmentation, skin inflammation, and skin aging.

  The active ingredient of the antioxidant function improver of the present invention is a δ-amino acid, a derivative thereof or a salt thereof. The δ-amino acids include the following formula (1)

(Wherein R ¹ represents a hydrogen atom or an acyl group, and R ² represents a hydrogen atom or an alkyl group which may have a substituent)
The 5-aminolevulinic acid represented by these or its salt is mentioned.
The 5-aminolevulinic acids are photosensitizers in photodynamic therapy (Japanese Patent Publication No. 2004-505105), plant growth regulators (JP 07-53487 A), herbicides (JP 05-117110 A), It is known that it is useful as a fish pathogenic microorganism, parasitic infection treatment (Japanese Patent Laid-Open No. 2001-316255), pig growth promoter (Japanese Patent Laid-Open No. 2003-40770), etc. Is not known at all.

In formula (1), examples of the acyl group represented by R ¹ include an alkanoyl group having 1 to 24 carbon atoms, an aromatic acyl group, and a benzyloxycarbonyl group. Specific examples of preferable acyl groups include acetyl group, n-propanoyl group, n-butanoyl group, n-pentanoyl group, n-hexanoyl group, n-nonanoyl group, benzyloxycarbonyl group and the like. Among these, a C1-C6 alkanoyl group is more preferable.

Moreover, as an alkyl group which may have a substitution shown by R < ² >, for example, a C1-C24 alkyl group which has a linear or branched or cyclic structure which may have a substituent Etc. Examples of the substituent that this alkyl group may have include a hydroxy group, an alkoxy group, and a phenyl group. Preferable examples of the alkyl group which may have such a substituent include, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-pentyl group, n-hexyl group, cyclohexyl group, n- Examples include a heptyl group, an n-octyl group, an n-nonyl group, an n-dodecyl group, an n-hexadecyl group, a benzyl group, a phonethyl group, a 3-phenylpropyl group, a hydroxyethyl group, and an ethoxyethyl group. Among these, a C1-C6 alkyl group is more preferable. R ¹ represents a substituent of an amino group, and R ² represents a substituent of a carboxylic acid group. These exemplified substituents are not only 5-aminolevulinic acid but also a δ-amino acid substituent. But there is.

  Examples of salts of δ-amino acids or derivatives thereof include hydrochloride, hydrobromide, hydroiodide, phosphate, nitrate, sulfate, acetate, propionate, toluenesulfonate, and succinic acid. Acid addition salts such as salt, oxalate, lactate, tartrate, glycolate, methanesulfonate, butyrate, valerate, citrate, fumarate, maleate, malate, and Examples thereof include metal salts such as sodium salt, potassium salt and calcium salt, ammonium salt, alkylammonium salt and the like. These salts are used as an aqueous solution or powder at the time of use.

  The above δ-amino acids, derivatives thereof or salts thereof may form hydrates or solvates, and any of them may be used alone or in combination of two or more.

  δ-amino acids, derivatives thereof or salts thereof (hereinafter referred to as “δ-amino acids”) can be produced by any method of chemical synthesis, production by microorganisms, and production by enzymes. Among the δ-amino acids or salts thereof, 5-aminolevulinic acid is disclosed in JP-A-48-92328, JP-A-62-111954, JP-A-2-76841, and JP-A-6-172281. It can be produced according to the method described in JP-A-7-188133 and JP-A-11-42083. The δ-amino acid or a salt thereof or a derivative thereof produced as described above, a chemical reaction solution or a fermentation solution before purification thereof can be used as they are without separation and purification as long as they do not contain harmful substances. . Moreover, a commercial item etc. can also be used.

  As will be described later in Examples, the δ-amino acids enhance the SOD activity or GPx activity of mice that are aged and have a decreased antioxidant function, and improve the decreased antioxidant function. Therefore, the above-mentioned δ-amino acids are useful as an antioxidant function improver for animals including humans, and are particularly useful as an antioxidant function improver for animals including persons whose antioxidant functions have decreased due to aging.

  Moreover, the antioxidant function improvement agent of this invention can improve the effect further by containing a mineral or ingesting simultaneously. Examples of the mineral include iron, zinc, copper, phosphorus, calcium, magnesium, potassium, selenium, chromium, manganese, iodine, boron, silicon, vanadium, molybdenum, cobalt, and the like, particularly preferably copper, zinc, manganese, Selenium. These mineral components can be used alone or in combination of two or more. Any chemical properties of minerals may be used as long as they do not harm the organism.

  The antioxidant function improver of the present invention can contain other antioxidants and, if necessary, nutrients. Examples of other antioxidants include polyphenols such as ubiquinone and ferulic acid, and flavonoids such as N-acetylcysteine, cysteine, catechol, tocopherol, catechin, and quercetin. Examples of nutrients include essential amino acids, non-essential amino acids, vitamins, herbs, proteins, various enzymes, and the like.

  The antioxidant function improver of the present invention comprises a powder of δ-amino acids, an aqueous solution in which δ-amino acids are dissolved in water, and a fermentation liquid containing δ-amino acids produced by the above method as a carrier such as an excipient. It can also be used after being adsorbed. As a kind of carrier, it may be a general one, crystalline cellulose, gelatin, starch, dextrin, oil cake, baker's yeast, brewer's yeast, liquor yeast, wine yeast, skim milk powder, lactose, animal and vegetable oils and fats, Anhydrous calcium phosphate, calcium carbonate, magnesium stearate, magnesium aluminate silicate, magnesium metasilicate aluminate and the like can be mentioned.

  Examples of the dosage form of the antioxidant function improver of the present invention include injections, tablets, capsules, fine granules, syrups, and suppositories. These can be produced according to a conventional method using a solvent, a dispersion medium, an extender, an excipient and the like as appropriate. Moreover, you may ingest as a form of food.

  When the antioxidant function improver of the present invention is prepared as an aqueous solution, when δ-amino acids are 5-aminolevulinic acid, in order to prevent decomposition of 5-aminolevulinic acid, its derivative or a salt thereof as an active ingredient, Care must be taken so that the aqueous solution does not become alkaline. When it becomes alkaline, decomposition of the active ingredient can be prevented by removing oxygen.

  The antioxidant function-improving agent of the present invention is not limited as long as the agent can be ingested to improve its antioxidant function, and the method of using the agent is not limited, but preferred embodiments are shown below.

  The target animals for the antioxidant function improver using this agent are not particularly limited, but vertebrates such as mammals, reptiles, birds, amphibians and fish are preferred. Examples of these include freshwater fish such as humans, cows, pigs, sheep, goats, mice, rats, rabbits, dogs, cats, chickens, salmon, rainbow trout, carp, eel, char, coho salmon, yellowtail, red sea bream, mackerel, tuna And saltwater fish.

  The use of the antioxidant function improver of the present agent is possible at any time during the growth of the animal, but preferably after the antioxidant function, for example, SOD activity or GPx activity is lowered. The use of the antioxidant function improver is preferably 15 years old or later, and particularly preferably 35 years old or later, for humans.

  The method for ingesting the antioxidant function-improving agent of the present invention is not particularly limited, and examples thereof include oral ingestion, ingestion or intestinal ingestion, and oral ingestion is particularly preferable.

  This drug shows a sufficient effect even if it is taken once, but it can be taken multiple times to further enhance the effect. The effect per ingested agent is more effective when taken multiple times, and it is an efficient usage method to take a small amount every day.

  The amount of intake of this drug per 1 kg of the target animal is preferably 0.001 mg to 1000 mg, more preferably 0.001 mg to 100 mg, and particularly preferably 0.001 mg to 50 mg. The intake of this drug should be increased as the period of growth increases and as the number of intakes decreases. Ingestion beyond the appropriate range is not desirable because it is not economical and can cause sun injury.

  Moreover, when using together minerals, you may use simultaneously and may use separately. The kind of mineral to be used, the method of using the same, and the amount of the mineral used may be the same as those of commercially available minerals. For example, in the case of copper, the amount used may be 0.5 to 10 mg per day for an adult male, with 1 to 5 mg being preferred. In the case of zinc, it may be 1 to 40 mg, and 5 to 20 mg is preferable. In the case of manganese, it may be 0.5 to 11 mg, preferably 2 to 8 mg. In the case of selenium, it may be 10 to 250 μg, and preferably 20 to 100 μg.

  EXAMPLES Next, although an Example is given and this invention is demonstrated further in detail, this invention is not limited to this at all.

Example 1
Mice preliminarily raised for one week (70 weeks old, BALB / cAJcl) were ingested with 10 mg of 5-aminolevulinic acid (ALA) hydrochloride per kg of mouse body weight once a day for 7 consecutive days. ALA hydrochloride was adjusted to a concentration of 0.5 mg / mL with distilled water and orally administered to mice. After the test, the mouse was dissected, the mouse thymus was collected, and the tissue superoxide dismutase (SOD) activity was measured. To the collected thymus, 1.0 mL of a 0.25M sucrose aqueous solution was added, and after homogenization, the supernatant obtained after centrifugation at 10,000 G for 10 minutes was used as an enzyme solution. The measurement was performed by the luminescence reagent MPEC (2-methyl-6-p-methoxyphenylethynylimidazopyrazinone; mw = 279.1) method (manufactured by Ato Co., Ltd.) for measuring antioxidant capacity. Specifically, 0.1 M potassium phosphate buffer pH 7.5, 125 μL, enzyme solution 10 μL, 0.1 units / mL xanthine oxidase 60 μL, distilled water 45 μL, 0.3 mM MPEC 10 μL, reaction final amount 250 μL, Luminescencer-PSN (Ato shares) And 50 μL of 0.72 mM hypoxanthine pH 7.5 was added to cause light emission. For the calculation of the luminescence inhibition rate, a buffer solution was used instead of the enzyme solution. For the calculation, a standard curve was prepared using a standard SOD, and the concentration was calculated from this.
In addition, glutathione peroxidase (GPx) activity was also measured. A 0.6 mL reaction mixture (containing 0.05 M K-phosphate buffer, pH 7.0, 1 mM NaN ₃ , 1 mM EDTA, 4 mM GSH) and 2 mM H ₂ O ₂ kept at 37 ° C. in the spectrophotometer cell. 0.1 mL and 50 μL of enzyme solution were added, and a total amount of 1 mL was directly scanned with a spectrophotometer at a wavelength of 340 nm (ODat 340 nm) for 2 minutes (20 mm / min). The empty sample used distilled water instead of the enzyme solution.
The calculation is performed by calculating the slope of the straight line ΔOD (ΔOD = Δcm (y-axis change)) × ODMax
/ Fullcm) using the NADPH molecular extinction coefficient a = 6.3 cm ² / μmole.

(Equation 1)
GPx activity = V / adv × ΔOD / Δt
(V is the amount of reaction solution, d = light path = 1 cm, v = enzyme amount, Δt = time change min)

Sought by.
The test was carried out with 5 male mice and 5 female mice per group, for a total of 10 mice, and the values showed average values. In the group treated with ALA, it was confirmed that SOD activity or GPx activity was improved in both male and female cases.

Claims (8)

The following formula (1)

(Wherein R ¹ represents a hydrogen atom, and R ² represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms)
The antioxidant function improvement agent which uses the compound or its salt represented by these as an active ingredient. The antioxidant function improving agent of Claim 1 which uses 5-aminolevulinic acid or its salt as an active ingredient.   Furthermore, the antioxidant function improving agent of Claim 1 or 2 which contains a mineral.   The antioxidant function improving agent according to claim 3, wherein the mineral is copper, zinc, manganese or selenium.   The antioxidant function improving agent according to any one of claims 1 to 4, which is an agent for oral, injection or enteral ingestion. The following formula (1)

(Wherein R ¹ represents a hydrogen atom, and R ² represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms)
In a compound or a salt thereof, an anti-oxidation function improving agent according to any one of claims 1 to 5 is also of you intake 0.001~1000mg per body weight 1kg per day. The following formula (1)

(Wherein R ¹ Represents a hydrogen atom, R ² Represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms)
The superoxide dismutase activity enhancer which uses the compound or its salt represented by these as an active ingredient. The following formula (1)

(Wherein R ¹ Represents a hydrogen atom, R ² Represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms)
The glutathione peroxidase activity enhancer which uses the compound or its salt represented by these as an active ingredient.