KR20070075077A - Composition comprising tyrosol derivatives for anti oxidation - Google Patents

Abstract

An antioxidation composition comprising tyrosol derivatives is provided to remove DPPH(diphenylpicrylhydrazyl) free radical and peroxide ion, and inhibit oxidation by TEAC(trolox-equivalent antioxidant capacity) method. The antioxidation composition for preventing and treating diseases associated with oxidation including cancer, aging, coronary arteriosclerosis, diabetes, epilepsy and neurodegenerative disease comprises 0.1 to 50 wt.% of a tyrosol derivative selected from para-tyrosyl gallate represented by the formula(1), meta-tyrosyl gallate represented by the formula(2) and ortho-tyrosyl gallate represented by the formula(3), wherein the composition is a pharmaceutical or health food composition having formulation of powder, granule, tablet, capsule or drink, and the daily dosage of the tyrosol derivatives is 0.01 mg/kg to 10 g/kg.

Description

Composition comprising tyrosol derivatives for anti oxidation

1 is a diagram showing the chemical structures of para-tyrosol, para-tyrosyl acetate, gallic acid and para-tyrosyl gallate,

2 is a diagram showing DPPH free radical scavenging activity against para-, meta-, ortho-tyrosyl gallate, para-tyrosyl acetate and gallic acid,

3 is a diagram showing the peroxide ion scavenging activity of para-, meta-, ortho-tyrosyl gallate, para-tyrosyl acetate and gallic acid,

4 is a diagram showing the antioxidant activity of para-tyrazole, para-tyrosyl acetate, para-tyrosyl gallate and gallic acid by TEAC assay.

The present invention relates to an antioxidant pharmaceutical composition and a dietary supplement comprising the tyrosol derivative compound.

Oxidative activity refers to an activity that prevents the production of free radicals in vivo and prevents oxidative phenomena that cause irreparable damage to cells. Steady state oxygen (triplet oxygen) is a superoxide radical, hydroxyl radical, hydrogen peroxide (H ₂ O ₂ ) due to environmental and biochemical factors such as enzymes, reduction metabolism, chemicals, pollutants, photochemical reactions, etc. It is known to convert into reactive oxygen species (ROS), such as ROS, to oxidize various cell components, such as lipids, proteins, and DNA, causing inflammation or damaging various organs (Beckman JS, et al., Proc) . ... Natl Acad Sci USA, 87, pp 1620 ~ 1624, 1990;. Sagar S, et al, Mol Cell Biochem, 111, pp 103 ~ 108, 1992;..... Ames BN, et al, Proc Natl . Acad, USA, 90, pp 7915 ~ 7922, 1993). Examples include superoxide anions (O ₂ ), hydrogen peroxide, hydroxyl radicals, singlet oxygen (IO ₂ ), alkoxyl radicals produced by lipid peroxidation, Peroxyl radicals (ROO) and nitrogen peroxide ions (ONOO-).

The action of these free radicals affects the action of antioxidant enzymes such as superoxide dismutase (SOD), catalase, and peroxidase, and vitamin C, E, and glutathione. However, if the biodefense is abnormal or exposed to excessive free radicals, the balance is broken, and the free radicals irreversibly destroy substrates, proteins, DNA, etc., and as a result, aging , Cancer, multiple atherosclerosis, arthritis and parkinson's disease have been reported to cause a variety of diseases (Griffiths HR, et al., FEBS Lett ., 388 , pp 161-164, 1996; Squadrito GL, et al., Free Radic . Biol . Med ., 25 , pp 392-493, 1998; Choi JS, Phytochem . Res., 16 , pp 232-235, 2002; Dreher D, et al., Eur . J. Cancer , 32A (1) , pp 30-38, 1996; Sohal RS, et al., Free Radic . Biol . Med ., 33 (1) , pp 37-44, 2002).

In most cases, the cells age before they become cancerous. Thus, many studies have suggested that aging and cancer may be holding both ends of the same spectrum (Shay JW et al., Radiat . Res. 155 , pp 188-193, 2001; Ershler WB et al., J. Natl . 89 , pp 1489-1497 , 1997; Campisi J., J. Am. Geriatr . Soc . 45 , pp 482-488 , 1997). The molecules that cause aging are very diverse, including environmental as well as genetic (Camplejohn RS et al., Br. J. Cancer 88 , pp 487-490, 2003). DNA must be replicated with high fidelity in order to transcribe accurate genetic information into daughter cells, and the decrease in activity and fidelity of DNA pol α has been shown to be an important part of genetic degeneration as an endogenous factor during aging. Several studies have shown a causal relationship between antioxidants and aging (Finkel T et al., Nature 408 , pp 239-247 , 2000; Srivastava VK et al., Experim . Gerontology 38 , pp 1285-1297 , 2003). . The mechanism of action of antioxidants includes the following three. (1) inhibits the production of free radicals by chelating enzymes or inhibitors associated with free radicals (2) removing free radicals (3) up-regulating antioxidant defenses (upregulating) or promoting. Free radicals, including superoxide anions, hydroxy radicals, singlet oxygen, and hydrogen peroxide, are produced in all mammalian cells from exposure to mitochondrial oxygen breathing or toxins. Some of these free radicals in vivo play a positive role in the production of energy and in the presentation of intracellular signaling pathways for apoptosis, stress, and proliferation or in the synthesis of biologically important components (Pietta PG., J Nat. Prod. 63 , pp 1035-1042, 2000). Intracellular defense mechanisms from free radicals include correction systems, detoxifying enzymes such as superoxide dismutases (SOD), and scavengers such as glutathione. When the generation of free radicals and their protection mechanisms are not balanced, it is suggested that the production of unnecessary free radicals is strongly linked to the processes of aging and degenerative diseases such as cardiovascular disease and cancer. In addition, free radicals can attack DNA by inducing cell membrane damage such as peroxidation of cell lipids and decreased fluidity of cell membranes through oxidation, which can also lead to denaturation of DNA that causes cancer (Burdon RH., Free Radic). ... Biol Med 18, pp 775 ~ 794, 1995; Russo A et al, Cell Biol Toxicol 16, pp 91 ~ 98, 2000;..... Sanchez-Moreno C., Food Res Int 32, pp 407 ~ 412, 1999). These observations suggest that free radicals may be an important target of preventive chemotherapy for aging-related diseases.

In order to prevent diseases caused by oxidative stress and prevent oxidative deterioration of foods, many studies are being conducted to isolate, identify and develop powerful and harmless natural antioxidants from edible or medicinal plants. Plant-derived natural compounds are also reported to retard oxidation by free radicals and scavengers or reducing agents of reactive oxygen species (Pratt, DE, American Chemical Society, pp 54-71, 1994: Larson, RA, Phytochem ., 27 , pp 969-978, 1988).

Synthetic antioxidants developed so far include butylated hydroxy anisole (BHA), butylated hydroxy toluene (BHT) and nordihydro-guaiaretic acid (NDGA). Antioxidant enzymes such as oxidase and non-enzymatic antioxidants such as tocopherol (vitamin E), vitamin C (vitamin C, ascorbic acid), carotenoids and glutathione.

Phenolic compounds, widely distributed in nature and considered the most representative antioxidants, are secondary metabolites with one or more hydroxyl groups that can be substituted in the ring structure. These compounds act as inhibitors and promoters for a variety of mammalian enzyme systems and as scavengers and metal chelators of oxygen radicals. Since this antioxidant activity is related to conjugated rings and hydroxyl groups, it is not surprising that most phenolic compounds have antioxidant activity (Sanchez-Moreno C et al., Food Res. Int . 32 , pp 407-412 , 1999; Decker EA., Nutr . Rev. 53 , pp 49-58 , 1995).

Para-tyrosol (p-Tyrosol, 4-hydroxyphenylethanol) is a well-known phenolic compound found in several dietary sources, such as virgin olive oil, wine, and sedum (Rhodiola species). .

Gallic acid, 3,4,5-trihydroxybenzoic acid, which is naturally abundant in plants, is also a well-known phenolic compound. It exists as a gallic acid in plants, but it is also a major by-product of gallotannin (Wiggins NL et al., J. Chem. Ecol . 29 , pp 1447-1464, 2003). Gallic acid not only has thrombolytic ability (Yun HS et al., Yakhak) Hoeji 37 , pp 453 ~ 457, 1993) It is known to prevent oxidative damage induced by free radicals and induce apoptosis of some tumor cells (HL-60RG, HeLa, KB cells, etc.) with better selectivity than normal cells. (Inoue M et al., Biochem . Biophys. Res. Commun . 204 , pp 898-904, 2002).

Excessive oxidative stress, a major cause of unnecessary free radical production, is strongly linked to degenerative diseases, including many types of cancer, and the aging process. Thus, in this research, the antioxidant activity of tyrosol and its derivatives was studied as part of the research and development efforts on anti-aging agents. To synthesize para-tyrosyl gallate combined with para-tyrosol and gallic acid and to investigate the relationship between structure and activity, meta- and ortho-tyrosyl gallate and para-tyrosyl acetate Also synthesized and studied. In this study, DPPH, NBT, and TEAC assays were performed to compare the radical scavenging ability of these compounds with vitamin C, a well-known antioxidant. We completed the present invention by confirming that para-tyrosyl gallate has high antioxidant activity in the tyrosol derivatives.

The present invention provides an antioxidant pharmaceutical composition comprising a tyrosol derivative compound exhibiting DPPH free radical scavenging activity, peroxide ion scavenging activity, and antioxidant activity.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of oxidation-related diseases comprising a tyrosol derivative compound.

The tyrosol derivative compound includes tyrosyl gallate and isomers thereof. The structural isomers also include para-tyrosyl gallate, meta-tyrosyl gallate or ortho-tyrosyl gallate represented by the following structural formulas (1) to (3).

The compound provides a pharmaceutical composition comprising 0.1 to 50% by weight of the compound based on the total weight of the composition.

Such oxidation-related diseases include cancer, aging, coronary atherosclerosis, diabetes, epilepsy, neurodegenerative diseases, preferably cancer, aging.

The present invention provides a health functional food comprising a tyrosol derivative compound selected from para-tyrosyl gallate, meta-tyrosyl gallate and ortho-tyrosyl gallate, which shows the effect of preventing and treating oxidative diseases. do.

The composition containing the tyrosol derivative compound of the present invention can be used in various ways, such as drugs, foods and beverages effective for preventing and improving oxidation-related diseases. In this case, the amount of the mixture in the food or beverage is generally added to the health food composition of the present invention 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably It can be added at a ratio of 0.3 to 1 g. Examples of the food to which the compound of the present invention may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

The tyrosol derivative compounds of the present invention may be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art. As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equivalent molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered. In this case, organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid, and methanesulfonic acid, p-toluene sulfonic acid, acetic acid, trifluoroacetic acid, and citric acid may be used as the organic acid. , Maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. Alkali metal or alkaline earth metal salts are obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the non-soluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salt, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the above compounds include salts of acidic or basic groups which may be present in the compound, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts. It can be prepared through.

The composition of the present invention preferably contains 0.01 to 99.9% of the tyrosol derivative compound, more preferably 0.1 to 90%. However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

Compositions comprising a tyrosol derivative compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The composition comprising the tyrosol derivative compound according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively. Carriers, excipients and diluents which may be formulated in a form and may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, Gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. Or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.

As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the tyrosol derivative compounds of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Since the tyrosol derivative compound itself of the present invention has almost no toxicity and side effects, it is a drug that can be used safely even when taken for a long time for the purpose of prevention and improvement.

The health beverage composition of the present invention, in addition to containing the compound as an essential ingredient in the indicated proportions, has no particular limitation on the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol.

As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Example  1. Preparation of Compounds

Tyrosol derivatives (p-, m-, and O-tyrosyl gallate and p-tyrosyl acatate) were synthesized from the laboratory of Prof. Bum-jong Lee of the Department of Biomedical Chemistry, Inje University, and para-, meta-, ortho-tyrosol ( Tyrosol, hydroxyphenethyl alcohol) and gallic acid were purchased from Aldrich (Aldrich. Chem. Co.) and used as samples in the following experimental example (FIG. 1). All samples were dissolved in ethanol and stored at 5 ° C. The samples were immediately diluted before all experiments were performed, and the final concentration of ethanol in each experiment did not exceed 1%. Other general reagents were commercially available Japanese-made express reagents.

Experimental Example  One. DPPH  freedom Radical  Scavenging activity measurement

1-1. Experiment preparation

This experiment makes it possible to measure the donating activity of hydrogen atoms (or single electrons) to determine the antioxidant activity of free radical scavenging. DPPH is a purple stable free radical that is reduced to yellow diphenyl picryl hydrazine. Various concentrations of tyrosol and its derivatives were reacted for 30 minutes at room temperature in a reaction mixture containing 0.1 mM DPPH-ethanol solution and 50 mM pH 7.4 Tris-HCl buffer 2: 1. By measuring the absorbance at 517 nm after the reaction, the degree of reduction of DPPH free radicals can be measured. Vitamin C was used as a positive control in this experiment and the inhibition rate (%) was calculated by the following equation:

% Inhibition = [(absorbance of control-absorbance of sample) / absorbance of control] x 100

1-2. Experiment result

The DPPH free radical scavenging experiment is one of the methods used to identify which substance can act as a free radical trap, which measures the conversion of DPPH free radicals to 2,2-diphenyl-1-picryl hydrazine by antioxidants. The results show the values obtained by performing three experiments. Para-, meta-, ortho-tyrazole did not show any radical scavenging activity even at the maximum concentration of 100 μM. However, para-, meta-, ortho-tyrosyl gallate showed excellent radical scavenging activity and the difference in activity according to the position of the phenolic hydroxyl group was not seen significantly (Fig. 2a).

As shown in FIG. 2B, para-tyrosyl acetate did not show any effect in the concentration range from 0.63 μM to 10 μM, and para-tyrosyl gallate and gallic acid showed relatively small differences and both had strong radical scavenging activity. Showed. Vitamin C showed 50% scavenging activity at 13.57 μM, whereas para-tyrosyl gallate and gallic acid showed about 50% scavenging at concentrations of 6.64 μM and 6.56 μM, respectively. These results show that para-tyrosyl gallate can have a strong free radical trap by the synthesis of para-tyrazole which did not show radical scavenging activity and gallic acid which showed strong scavenging activity. And that if the phenol with two or more hydroxyl groups relative to phenolic (phenol) with the hydroxyl groups of less than one or more strong antioxidant hypothesis that has the activity (Rice-Evans CA et al. , Free Radical Biol. Med . 20, pp 933 ~ 956, 1996) and the support.

Experimental Example  2. NBT  method( nitroblue - tetrazolium  assay)

2-1. Experiment preparation

This experiment is the result of competition between the sample and NB for peroxide anions produced by the xanthine-xanthine oxidase system. The change in absorbance of NBT over time was observed. Inhibition percentage for the rate values obtained from the control without samples is indicated by the relative peroxide anion scavenging ability of these samples. Vitamin C was used as a positive control. Reagents for this experiment were prepared at 50 ° C. in a 50 mM KH ₂ PO ₄ / KOH buffer (pH 7.4) at 25 ° C. and the reaction mixture was 20 μl of 15 mM Na ₂ EDTA, 50 μl of 0.6 mM NBT, Each sample was dissolved in 30 μl of 3 mM hypoxanthine and 3 μl of ethanol and the reaction was carried out by 50 μl of xanthine oxidase (1 unit / 10 ml buffer) at 25 ° C. Started. The absorbance was measured at 570 nm for 10 minutes at 30 second intervals using a Multi Microplate Reader (synergy HT, BIO-TEK®). The control group uses the same amount of sample solvent ethanol instead of the sample and the result is expressed as a relative inhibition rate (%) relative to the control group was calculated by the following equation:

% Inhibition = [(control response-sample response) / control response] x 100

2-2. Experiment result

Peroxide ions and hydroxyl radicals are the most representative free radicals. In an intracellular oxidation reaction, peroxide ions are usually formed first, which can increase the effects of these peroxide ions because they produce oxides and other harmful free radicals. Xanthine oxidase is one of the main enzymatic causes of these free radicals in vivo. In the NBT method, hypoxanthine-xanthine oxidase generates peroxide ions, and the peroxide ions reduce NBT to form a blue forazan. Tyrosol and its derivatives inhibited the reduction of NBT by hypoxanthine-xanthine oxidase in a concentration dependent manner (FIGS. 3A and 3B).

At concentrations of 0.625 μM, para-tyrosyl gallate and gallic acid showed about 50% inhibition, but vitamin C and para-tyrosyl acetate showed no inhibition at all concentrations (0.625-20 μM) in this experiment. It was not shown (FIG. 3A). In particular, unlike other antioxidant activity experiments, this NBT method showed a slight difference in antioxidant activity of para-, meta-, ortho-tyrosyl gallate depending on the position of the hydroxyl group attached to the phenol group (FIG. 3b). 0.625 μM of para-tyrosyl gallate showed 12% higher scavenging activity than meta-tyrosyl gallate and 8% higher than ortho-tyrosyl gallate. However, para-, meta- and ortho-tyrosol did not show any peroxide ion scavenging activity as the result of DPPH assay.

Experimental Example  3. TEAC  Measurement of values

3-1. Experiment preparation

Determination of the trolox equivalent antioxidant capacities (TEAC) values for tyrosol and its derivatives was performed according to methods that have been studied in the art (Re R et al., Free Radic . Biol . Med . 26 , pp 1231-1237 , 1999). ABTS. + (ABTS radical cation) is produced by the reaction of 2.45 mM potassium persulfate with ABTS solution dissolved in water at a concentration of 7 mM and prepared by placing it in a dark place at room temperature 12 hours before use. . The ABTS. + Solution was diluted with 5 mM PBS to a absorbance value of 0.70 ± 0.02 at 734 nm, and then 10 μl of sample or standard trolox was added to 1 ml of this ABTS. + Solution. After reacting at 30 ° C. for 6 minutes, absorbance was measured at 734 nm. Vitamin C was used as a positive control, and the antioxidant activity of tyrosol and its derivatives was expressed as TEAC value using standard curves calculated by different concentrations of trolox.

3-2. Experiment result

The TEAC method by the generation of ABTS radicals is one of the spectrophotometric methods that have been used to measure the total antioxidant activity of a sample. In the same experimental state, the antioxidant activity of the sample was determined by comparing the scavenging reactivity of the ABTS radical (2,2 -azinobis (3-ethylbenzo-thiazoline-6-sulfonic acid) of the standard trolox antioxidants in%. All compounds in the TEAC experiment showed ABTS radical scavenging activity in a concentration dependent manner (FIG. 4) Para-tyrosol showed the highest TEAC value in gallic acid and no noticeable antioxidant activity in other experiments. And para-tyrosyl acetate showed higher TEAC values than vitamin C. This may be because para-tyrosol and para-tyrosyl acetate have better binding to ABTS radicals than vitamin C.

The results of this experiment showed that para-tyrosyl gallate had about three times stronger antioxidant activity than trolox. Para-, meta-, ortho-tyrosyl gallate showed the same TEAC value.

Formulation example  One. Powder  Produce

p-tyrosyl gallate ........................ 20 mg

Lactose ......................................... 100 mg

Talc ........................................ 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

m-tyrosyl gallate .................................. 10 mg

Corn starch ........................ 100 mg

Lactose ......................................... 100 mg

Magnesium Stearate ......................................... 2 mg

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation example  3. Manufacture of capsule

o-tyrosyl gallate .................................. 10 mg

Crystalline Cellulose ......................................... 3 mg

Lactose ............... 14.8 mg

Magnesium Stearate ............ 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation example  4. Preparation of Injectables

p-tyrosyl gallate ......................................... 10 mg

Mannitol ......................................... 180 mg

Sterile distilled water for injection ........................ 2974 mg

Na ₂ HPO _{4 12} H ₂ O ... 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation example  5. Liquid  Produce

m-tyrosyl gallate .................................. 20 mg

Isomerized sugar ......................................... 10 g

Mannitol ......................................... 5 g

Purified water ...............

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of health food

o-tyrosyl gallate ..................... 1000 mg

Vitamin Blend ........................

Vitamin A Acetate ......... 70 μg

Vitamin E ........................ 1.0 mg

Vitamin B1 ........................ 0.13 mg

Vitamin B2 ........... 0.15 mg

Vitamin B6 ............... 0.5 mg

Vitamin B12 ......................... 0.2 μg

Vitamin C ............... 10 mg

Biotin ......................................... 10 μg

Nicotinic acid amide ......... 1.7 mg

Folic acid ......................... 50 μg

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture ........................

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium Carbonate ............... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dicalcium Phosphate Dibasic ............... 55 mg

Potassium Citrate ... 90 mg

Calcium Carbonate ... 100 mg

Magnesium Chloride ............... 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

p-tyrosyl gallate ........... 100 mg

Vitamin C ......................................... 15 g

Vitamin E (powder) ..................... 100 g

Iron Lactate ............... 19.75 g

Zinc Oxide ......................... 3.5 g

Nicotinic acid amide ...................... 3.5 g

Vitamin A ......................... 0.2 g

Vitamin B1 ......... 0.25 g

Vitamin B2 ............. 0.3g

Water ............................................

After mixing the above components in accordance with the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare a healthy beverage composition of the invention.

Although the composition ratio is mixed with a component suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As mentioned above, the tyrosol derivative compound of the present invention exhibits DPPH free radical scavenging activity, peroxide ion scavenging activity, and antioxidant activity by TEAC method, and thus can be usefully used as a more effective and powerful antioxidant composition. have.

Claims (6)

A pharmaceutical composition for the prevention and treatment of oxidative related diseases comprising a tyrosol derivative compound selected from para-tyrosyl gallate, meta-tyrosyl gallate and ortho-tyrosyl gallate. The pharmaceutical composition of claim 1, wherein the compound comprises 0.1 to 50% by weight of the compound, based on the total weight of the composition. The pharmaceutical composition of claim 1, wherein the oxidation-related disease is cancer, aging, coronary atherosclerosis, diabetes, epilepsy, or neurodegenerative disease. The pharmaceutical composition according to claim 3, wherein the oxidation-related disease is cancer or aging. A health functional food comprising a tyrosol derivative compound selected from para-tyrosyl gallate, meta-tyrosyl gallate, and ortho-tyrosyl gallate, which shows the effect of preventing and treating oxidation-related diseases. A dietary supplement according to claim 5 which is a powder, granule, tablet, capsule or beverage.

Images