KR20010079464A - Antioxidant composition comprising anti-aging preparation - Google Patents

Abstract

PURPOSE: An antioxidant composition comprising lycopene, vitamin C, vitamin E(alpha-tocopherol), coenzyme Q10 and lipoic acid is provided, which has excellent aging preventing effect higher than that of conventional antioxidant compositions. CONSTITUTION: This antioxidant composition having excellent ageing preventing effect contains 3 to 20 parts by weight of lycopene, 100 to 1,000 parts by weight of vitamin C, 100 to 800 parts by weight of vitamin E(alpha-tocopherol), 5 to 80 parts by weight of coenzyme Q10 and 5 to 100 parts by weight of lipoic acid. The effective daily intake amount of the composition is orally 700 to 1,300mg as tablets.

Description

Antioxidant composition comprising anti-aging preparation

The present invention relates to five anti-aging natural antioxidant formulations having different redox potentials, and more specifically, known antioxidants such as lycopene, vitamin C (L-ascorbic acid) and vitamin E (α-tocopherol). ), And coenzyme Q ₁₀ and lipoic acid (lipoic acid) in a suitable composition to the antioxidant formulation composition useful for anti-aging.

In addition, the anti-aging antioxidant formulation composition of the present invention prevents cell aging due to free radicals generated in the body and has a great effect in preventing skin aging due to ultraviolet rays.

Lycopene, vitamin C (L-ascorbic acid), vitamin E (α-tocopherol), coenzyme Q ₁₀ and lipoic acid, which are components of the present invention, are already known as antioxidants. However, these antioxidant effects are known only for the antioxidant effect of each component, and when formulated to an appropriate composition, the anti-aging effect due to the synergistic effect of each antioxidant component is not yet known.

In order to maintain physical and mental health, various essential nutrients, compounds such as coenzyme and cofactor should be properly ingested in the body, and good air should be inhaled by the human body. Oxygen in the air is essential for supplying nutrients by oxidizing various carbohydrates, proteins and fats in the body.

Aging is generally promoted by disruption of metabolic balance in cells, incomplete oxidation or lack of metabolic / redox intermediates, and the like, which also cause various diseases such as cancer, heart disease, mental illness or skin aging.

Generally, reactive oxygen species (ROS), such as superoxide radicals, hydroxyl radicals or hydrogen peroxide, arise from incomplete oxidative metabolites from intracellular redox reactions or from mitochondrial electron delivery systems. Insufficient levels of reducing agents such as NADH, succinate and the like promote the formation of poor ROS.

Such reactive free radicals are often the cause of cancer or heart disease, and minimizing the level of ROS is very important for maintaining a healthy body and preventing skin aging. Various antioxidants are required for the reduction of ROS, and vitamin C, E or various cofactors are known to be useful as antioxidants.

Representative antioxidants include lycopene, which is a carotenoid pigment in tomato, grapefruit or watermelon. Lycopene is known to be useful for the prevention of heart disease or prostate cancer. It is also useful for the prevention of atherosclerosis.

Vitamin C is an antioxidant widely used in various foods and is known to be useful for the prevention and treatment of colds, various cancers, and heart diseases. It is also known to be useful for strengthening arteries in adults after middle age and for the prevention of cataracts.

Vitamin E is generally divided into four types, which can be divided into alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol and the like. In general, alpha-tocopherol is a major component of vitamin E. Vitamin E is useful as an anticoagulant and is also a powerful antioxidant. Vitamin E is also effective in preventing cancer, heart disease, cataracts, and atherosclerosis. Recent studies have shown that vitamin E can reduce the responsiveness of heart attacks to less than one-third and it is clear that it prevents the oxidation of low-density cholesterol (LDL), or bad cholesterol.

Oxidation of LDL leads to the formation of lipid plaques on the artery walls, leading to atherosclerosis. Therefore, vitamin E not only prevents atherosclerosis, but recent studies have shown that it is effective in boosting immunity and helps prevent Alzheimer's disease.

In addition, coenzyme Q (ubiquinone) is coenzyme Q in the cytochrome reductases of NADH-Q reductase, cytochrome reductase and cytochrome oxidase, which are three protein complexes used in the energy-producing pathway in the body mitochondria. Accept electrons through ₁₀ or ubiquinone. Therefore, the supply of ubiquinone is very important for high energy demanding cells such as the heart. Coenzyme Q ₁₀ is also useful as an antioxidant that prevents cell death caused by ROS as it ages. Ubiquinone plays a very important role in intracellular metabolism because the lack of these substances leads to the leakage of electrons from the mitochondrial electron transport system, leading to the formation of undesirable superoxides and other ROS.

Lipoic acid is a substitutive family of lipoic acid transacetylases, one of the components of the paruribate dehydrogenase complex that plays a very important role in operating the citric acid cycle during body metabolism. Therefore, it is involved in the hydrogen carrier and succinyl transfer, and lipoic acid is a catalyst between the formation of the acyl group and the transition reaction performed by these complexes. In addition, lipoic acid is considered to be useful for the prevention of heart disease, shock due to diabetes, cataracts, etc. and for the treatment of chronic diabetes including neurological damage.

Therefore, lycopene, vitamin C (L-ascorbic acid), vitamin E (α-tocopherol), coenzyme Q ₁₀ and lipoic acid, which are components of the present invention, all exhibit important physiological activities in the human body. Already known, they are all known to have antioxidant effects.

However, it is not yet known whether these five ingredients together or taken together are more effective than anticipated antioxidant effects.

The technical problem to be achieved by the present invention is lycopene (lycopene), vitamin C (L- ascorbic acid), vitamin E (α- tocopherol), coenzyme Q ₁₀ and lipoic acid (lipoic acid) when ingested in an appropriate ratio The present invention has been completed by discovering that the anti-aging effect is superior to the anti-aging effect by the antioxidant effect.

Accordingly, an object of the present invention is to provide five anti-aging natural antioxidant composition having different redox potential, more specifically 3 to 20 parts by weight of lycopene, 100 to 1000 parts by weight of L-ascorbic acid, α-tocopherol An anti-aging nutritional drug formulation containing 100 to 800 parts by weight, 5 to 80 parts by weight of coenzyme Q _{10 and} 5 to 100 parts by weight of lipoic acid is provided.

More preferably, 7 to 13 parts by weight of lycopene, 400 to 600 parts by weight of L-ascorbic acid, 300 to 500 parts by weight of α-tocopherol, _{10 to} 60 parts by weight of coenzyme Q and 50 to 100 parts by weight of lipoic acid are contained. It is to provide an anti-aging nutritional drug formulation.

The daily dose of the preparation of the present invention is preferably 700 to 1300 mg of the preparation when ingested in the form of a tablet.

Hereinafter, the present invention will be described in more detail.

The present invention relates to antioxidants that inhibit harmful free radicals, including free radicals, which are known to cause aging phenomena (skin wrinkles, physical health weakness, immune decline and memory loss, etc.). It relates to the preparation of.

Bioenzymatic reactions to sustain life occur cooperatively. Therefore, the synergistic action of vitamins and cofactors directly required for enzyme reactions ensures that the biofunction functions normally. If only one of the ten necessary vitamins and cofactors is missing, the pathway or cycle of the bioreaction (for example, the glycolytic pathway of carbohydrate metabolism and the citric acid cycle that oxidizes all three nutrients) loses the co-operation of the life-supporting enzyme reaction The reaction is stopped.

For this reason, vitamins and cofactors can be taken as multivitamin tablets instead of separately to lubricate the coordination of bioreactions. Based on the cooperative effects of the living body, the anti-aging agent has invented five antioxidants based on recent scientific results to prevent the aging phenomenon.

The present anti-aging antioxidant compositions are constructed as follows according to the rank of their redox potentials.

Lycopene-1.0 Volt Coenzyme Q10 (Ubiquinone) + 0.10 Volts Vitamin C + 0.08 Vitamin E-0 ∼ Lipoic acid-0.29 〃

* Reduction potential of lycopene cation radical ～ 1000mV

(Mortenson et al, 2001)

As seen from the redox potential above, the formulation contains a wide range of antioxidants from +1 Volt of lycopene to -0.29 Volt of lipoic acid. Therefore, free radicals generated in ROS and other various in vivo can be efficiently processed. For example, ROS O ₂ , OH and H ₂ O ₂ with different redox potentials can be synergistically treated with antioxidants with a wide redox potential range.

In particular, by treating the cationic radicals resulting from the antioxidant of lycopene in the order shown above (lycopene → coenzyme Q10 → vitamin C → vitamin E → lypoic acid) can exhibit a synergistic effect and anti-aging efficacy can be expected.

Hereinafter, the present invention will be described in detail through examples. However, these examples do not limit the scope of the present invention.

(Production Examples 1 to 3)

To prepare an antioxidant formulation having the composition of Table 1. Antioxidant agents can be added to the appropriate form of excipients for animal experiments and the active ingredients of each composition are shown in Table 1. The antioxidant preparations prepared in each preparation example were named S1, S2, S3.

S1 S2 S3 Lycopene 10 mg 15 mg 5 mg L-ascorbic acid 500 mg 500 mg 500 mg α-tocopherol 400 I.U. 200 I.U. 600 I.U. Coenzyme Q ₁₀ 30 mg 10 mg 60 mg Lipoic acid 50 mg 80 mg 10 mg

(Comparative Examples 1 to 5)

An antioxidant formulation having the composition of Table 2 was prepared. Antioxidant agents can be added to the appropriate form of excipients for animal experiments and the active ingredients of each composition are shown in Table 2. The antioxidant preparations prepared in each of the comparative examples were named C1, C2, C3, C4, C5.

C1 C2 C3 C4 C5 Lycopene - - 5 mg - 10 mg L-ascorbic acid 510 mg 525 mg 510 mg 550 mg 510 mg α-tocopherol 400 I.U. 200 I.U. - - - Coenzyme Q ₁₀ 30 mg - 60 mg - 30 mg Lipoic acid 50 mg 80 mg - 50 mg 50 mg

As shown in Table 2, the L-ascorbic acid content was added by the amount of the remaining antioxidant component removed so that the content of the overall antioxidant formulation was similar to that of the formulation prepared in the preparation examples.

Example 1 Invitro Free Radical Scavenging Experiment

The free radical scavenging ability of the formulations of Preparation Examples and Comparative Examples was measured as follows. The method used was carried out by the 1,1-Diphenyl-2-Pycryl-Hydrazyl (DPPH) method (Blois, M.S. Nature 181, 1190 (1958)). DPPH is a relatively stable free radical which, when present in the radical state, exhibits maximum absorption at 517 nm and loses its absorbance when erased. DPPH was used by Sigma Co., Ltd. and dissolved in methyl alcohol at a concentration of 0.15 mM. First, the preparation of Preparation Examples and Comparative Examples was prepared in 10 ㎍ / ㎖ 100 ㎕ to each well of the 96 well plate. 100 μl of DPPH solution was added thereto, and the mixture was left at room temperature for 30 minutes, and then absorbance at 517 nm was measured using a Micro plate reader (BioTek EL-340).

In this case, only 100 μl of methyl alcohol was used as a control to evaluate the antioxidant properties of the Preparation Examples and Comparative Preparation Examples. Preparation Example and Preparation Example The concentration of the preparation example and preparation example when the absorbance of the sample of the preparation was half the absorbance of the control group was expressed as IC ₅₀ (Inhibitory Concentration 50). Is shown in Table 3 below.

Free radical scavenging capacity of the preparation sample (unit: w / v%) S1 S2 S3 C1 C2 C3 C4 C5 IC ₅₀ at 30 min 0.0030 0.0027 0.0032 0.0015 0.0012 0.0017 0.0009 0.0011 IC ₅₀ at 60 min 0.0028 0.0025 0.0030 0.0010 0.0009 0.0015 0.0008 0.0010 IC ₅₀ at 12 hr 0.0020 0.0015 0.0021 0.0007 0.0006 0.0010 0.0006 0.0008

As shown in Table 3, it can be seen that the samples prepared in the preparation examples maintain free radical scavenging ability of about 2 times or more than the samples prepared in the preparation comparative example.

Example 2 Inhibition of Production of Intracellular Active Oxygen Induced by Hydrogen Peroxide

Preparation Examples and Comparative Preparation Examples The following experiments were carried out to investigate the effect of inhibiting intracellular free radicals induced by hydrogen peroxide of the preparation. The degree of oxidation by ROS was measured using H2DCFDA (2 ', 7'-dichlorodihydrofluorescein diacetate, Molecular Probes, Eugene, OR, USA). H2DCFDA, which is nonpolar, changes into membrane impermeable H2DCF (2 ', 7'-dichlorodihydrofluorescein) by intracellular esterases through the cell membrane. Since H2DCF is changed to 2 ', 7'-dichlorofluorescein (DCF), which is low fluorescence due to low fluorescence or intracellular active oxygen, the degree of oxidation of H2DCFDA can be known from the degree of generation of DCF. The cells used in the experiments were endothelial HUVECs (ATCC), which is endothelial cells, or rat thoracic aorta smooth muscle cell lines (ATCC), A7r5, which is a vascular smooth muscle cell. HUVEC was supplemented with A7r5 in Kaighn's F12K medium containing 10% FBS (fetal bovine serum), heparin sodium salt (0.1 mg / ml), endothelial cell growth aid (0.03-0.05 mg / ml) and 1% antibiotic. Was cultured using DMEM (Dulbecco's Modified Eagle's Medium) medium containing 10% FBS (fetal bovine serum) and 1% antibiotic.

After removing the medium of the cell monolayer, preparative Examples and Preparation Examples (10 ^-6 M) for 30 minutes at 37 ℃ and added to H ₂ O ₂ to the final concentration of 10 ^-5 M 20 minutes Treated. After blocking the light for 2 hours at 37 ° C. in 50 mM phosphate buffer containing 5 μM of H2DCFDA, the degree of oxidation of H2DCFDA was measured by a fluorescence analyzer (Fluorescence reader, FL600, Biotek Instruments) (485 nm excitation, 530 nmemission). ). The experimental results are shown in Table 4 below.

Inhibitory effect of the production of free radicals induced by hydrogen peroxide Test formulation cell % Inhibition cell % Inhibition S1 HUVEC 67.4 A7r5 66.5 S2 70.3 67.4 S3 71.4 72.4 C1 49.5 59.7 C2 55.4 67.0 C3 59.8 60.4 C4 60.4 61.5 C5 58.4 54.9

As shown in Table 4, the preparations prepared in the preparation example inhibited the generation of free radicals in HUVEC cells induced by hydrogen peroxide better than the preparations prepared in the preparation comparison example. In addition, A7r5 cells in the preparation of the preparation example showed a free radical inhibition than the preparation of the comparative example.

Example 3 Inhibition of Lipid Peroxidation in Biofilm

Preparation Examples and Comparative Examples Microsomes isolated from livers of rats were used as biofilms for measuring the lipid peroxidation inhibitory activity of the preparations. Microsomes were separated by fractional centrifugation. That is, the livers of the rats were extracted, washed in 2-3 M sucrose solution at 4 ° C., washed 2-3 times, and then chopped. Then, the sucrose solution of 8 times the weight of the liver, 5 mM tris-hydrochloric acid buffer solution ( pH 7.4) and 0.1 mM EDTA mixed solution were added and ground using a tissue grinder. This was centrifuged at 7,000 g for 10 minutes, the supernatant was taken, ultracentrifuged at 77,000 g for 60 minutes, and the precipitate was taken up and washed three times with Tris-HCl buffer. The suspension was diluted with Tris-hydrochloric acid buffer solution and the protein was quantified by Lowry method to 1 mg / ml. Lipid peroxidation inhibitory activity of the samples was measured using the isolated liver macrosomes of rats as biofilm lipid sources. The reaction was initiated by adding 10 μl of microsome solution, 10 μl of sample in each concentration dissolved in methanol, 0.5 ml of 100 mM Tris-hydrochloric acid buffer (pH 7.5), 450 μl of distilled water, and 30 μl of 4mM FeSO ₄ solution. . The reaction was carried out for 30 minutes in a 37 ° C. water bath, and 0.3 ml of 3M trichloroacetic acid-2N hydrochloric acid 1: 1 mixture was added to stop the reaction. After centrifugation at 3000 g for 10 minutes, 1 ml of the supernatant was added, and 0.3 ml of 0.67% thiobarbituric acid (TBA) was added to react for 10 minutes in a boiling water bath. After measuring the absorbance at a wavelength of 530 nm using a spectrophotometer, the lipid peroxidation inhibitory activity (%) was calculated by the following formula.

% Inhibition = (A-C) / (A-B)

A: absorbance without the inhibitor

B: absorbance without inhibitor and microsome

C: absorbance of the inhibitor

As a result, Table 5 shows the lipid peroxidation inhibitory activity of the Preparation Examples and Comparative Preparation Preparations with the concentration of 50% inhibiting lipid peroxidation (IC ₅₀ ) of the microsomes extracted from the liver of rats.

Biofilm Lipid Peroxidation Inhibition Test formulation IC ₅₀ Test formulation IC ₅₀ S1 0.8 μg / ml C2 1.5 μg / ml S2 0.9 μg / ml C3 1.4 μg / ml S3 0.7 μg / ml C4 1.6 μg / ml C1 1.1 μg / ml C5 1.3 μg / ml

Through such an embodiment, it can be seen that the anti-aging agent of the present invention has an excellent antioxidant effect as compared to conventional antioxidants. Therefore, the nutritional drug formulation consisting of the five antioxidant components of the present invention can significantly lower the content of free radicals in the body, which is sufficient to exhibit an anti-aging effect when consumed by the general public.

The combination of these five antioxidants shows higher antioxidant effects compared to conventional antioxidants alone or in combination, characterized by the combination of vitamin C, lycopene and α-tocopherol, in combination with coenzyme Q ₁₀ and lipoic acid. Due to its antioxidant and anti-aging effects will be seen to be maximized.

The effects of the present invention can be achieved by lycopene, vitamin C (L-ascorbic acid), vitamin E (α-tocopherol), coenzyme Q ₁₀ and lipoic acid, when ingested at an appropriate ratio. It is to provide a formulation having an anti-aging effect even better than the anti-aging effect.

Claims (3)

Lycopene 3 to 20 parts by weight of L- ascorbic acid 100 to 1000 parts by weight of α- tocopherol 100-800 parts by weight of coenzyme Q ₁₀ and lipoic acid 5 to 100 parts by weight of 5~80 anti-aging nutritional pharmaceutical preparation containing parts by weight According to claim 1, containing 7 to 13 parts by weight of lycopene, 400 to 600 parts by weight of L-ascorbic acid, 300 to 500 parts by weight of α-tocopherol, _{10 to} 60 parts by weight of coenzyme Q and 50 to 100 parts by weight of lipoic acid. Anti-aging nutritional drug formulations The anti-aging nutritional drug formulation according to claim 1, wherein the daily adult dosage of the anti-aging nutritional drug formulation is 700 to 1300 mg when taken in the form of a tablet.