KR100759468B1 - Composition for prevention and treatment of anti-gout comprising purple pigments isolated from purple sweet-potato - Google Patents

Abstract

A composition comprising an ethanol extract of purple sweet potato or an anthocyanin-based purple pigment isolated therefrom is provided to decrease the blood uric acid, a main causing material of gout, and inhibit the expression of gelatinase B induced by uric acid crystals. A composition or health food for preventing and treating gout comprises an anthocyanin-based purple pigment represented by the formula(1) as an effective ingredient. In the formula(1), R1 is a group represented by the structural formula(1-a), and R2 is a group represented by the structural formula(1-b), (1-c) or (1-d).

Description

Composition for prevention and treatment of anti-gout comprising purple pigments isolated from purple sweet-potato}

1 is a result of confirming gelatinase B expression by zymography after treatment for each concentration of purple pigment.

2 is a diagram showing the decrease in blood uric acid in purple pigment-treated mice.

The present invention relates to a composition for preventing and treating gout containing a purple pigment separated from purple sweet potatoes, and more particularly, a purple sweet potato ethanol extract or purple pigment isolated therefrom is used as an enzyme that is important for the formation of uric acid. It can be used as a material for medicines and health foods by inhibiting the expression of gelatinase B, which is highly expressed in patients with gouty arthritis, by inhibiting blood uric acid, thereby reducing blood uric acid. A new medicinal use of sweet potato ethanol extract or purple pigment.

Pigments present in many fruits, leaves, stems, and flowers of many plants have been suggested as potential sources of pigments for anthocyanins. Among them, many pigments such as cranberries, strawberries, cherries, carrots, red potatoes, purple sweet potatoes, etc. Research has been done. The purple sweet potato, which has recently attracted attention as a new natural food coloring source, was cultivated by introducing a variety known as Sancheonja, which was native to Kyushu, Japan. The original purple sweet potato has a normal yield of sweet potato. Compared to about 70% and the pigment content is not high, it is known to have low economic value, but recently, breeding has been developed excellent in terms of unit production and pigment content, so that it can be developed as a new natural food coloring source. It is expected to be used as a raw material for various processed foods. Purple sweet potatoes are dark purple not only in the epidermis but also in the flesh.

The purple sweet potato, which has recently been attracting attention as a new natural food coloring source, is a cultivated variety introduced in Korea, known as a wild-spring, which is native to Kyushu. Purple sweet potato is not only easy to cultivate and inexpensive, but also has a deep purple color of epidermis and flesh, and is known to be higher than other pigment sources. Also, anthocyanin pigment of purple sweet potato is peonydin ( The basic structure of peonidin) has a structure in which dicylation of ferulic acid and caffeic acid is formulated (formula 1), and thus it is highly available as a pigment in a very stable form.

이고, R₂는

,In Formula 1, R ₁

R ₂ is

,

또는

이다.

or

to be.

Gout is a term used to describe a group of diseases that are found only in humans. Monosodium urate crystals in synovial fluid leukocytes with symptoms of increased serum uric acid, recurrent acute arthritis. The presence and presence of monosodium urate crystals (tophi) in and around the joints, which sometimes leads to joint malformation and disability, accompanied by kidney disease, and manifests in different combinations.

Hyperuricemia is an increase in the concentration of uric acid in the blood. This occurs when the concentration of monosodium urate in serum exceeds the limit of solubility. At 37 ° C., the uric acid saturation of plasma is about 7 mg / dl. Therefore, if this concentration is exceeded, it becomes physicochemically supersaturated. Serum uric acid concentrations are known to be relatively high when they exceed +2 SD above the average serum uric acid concentration of normal subjects. In most epidemiological studies, the upper limit for men is 7 mg / dL and for women 6 mg / d. For this reason, the substantial upper limit for hyperuricemia is defined as more than 7.0 mg / dL. Gout is called "King of Diseases" and "Kings of Kings", a disease that occurs more often with age, and is accompanied by tremendous pain caused by inflammatory reactions caused by uric acid crystals of the big toe. Drugs having undesired side effects are being used as therapeutic agents, and thus, there is a need for the development of a substance that inhibits the expression of gelatinase B, a protease that is caused by monosodium uric acid crystals and is involved in tissue destruction while having no side effects.

Accordingly, the present inventors, while searching for a substance having a gout treatment effect from a plant material, the purple sweet potato ethanol extract or an anthocyanin-based purple pigment isolated therefrom reduces uric acid in the blood while reducing blood uric acid, which is the main source of gout. The present invention was completed by confirming the dual effect of inhibiting the expression of gelatinase B, a protease induced by the present invention.

[Formula 1]

이고, R₂는

,In Formula 1, R ₁

R ₂ is

,

또는

이다.

or

to be.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing and treating gout containing purple sweet potato ethanol extract as an active ingredient.

It is also an object of the present invention to provide a health food for gout prevention and treatment containing purple sweet potato ethanol extract as an active ingredient.

The present invention is characterized by a pharmaceutical composition for preventing and treating gout containing purple sweet potato ethanol extract as an active ingredient.

In addition, gout preventive and therapeutic health foods containing purple sweet potato ethanol extract as an active ingredient.

Referring to the present invention in more detail as follows.

In the present invention, purple sweet potato ethanol extract or purple pigment isolated therefrom inhibits acidic oxidase which is an important enzyme for the formation of uric acid, thereby reducing blood uric acid, thus exhibiting anti-inflammatory action without side effects and being highly expressed in patients with gouty arthritis. The present invention relates to a new medicinal use of purple sweet potato ethanol extract or purple pigment, which can be usefully used as a material for medicines and health foods by exhibiting a gout prevention and treatment action that suppresses the expression of.

First, the preparation method of the compound represented by Formula 1 according to the present invention

1) extracting the fine purple sweet potato with ethanol to obtain an ethanol extract;

2) filtering the ethanol extract with a membrane filter and then completely removing the ethanol with a vacuum evaporator;

3) performing column chromatography on the residue remaining after ethanol removal, adsorbing anthocyanin-based violet pigment and eluting with ethanol;

4) concentrating the eluate under reduced pressure to obtain an anthocyanin violet pigment.

The anthocyanin-based purple pigment thus separated is a compound represented by the following formula (1).

[Formula 1]

이고, R₂는

,In Formula 1, R ₁

R ₂ is

,

또는

이다.

or

to be.

The purple sweet potato ethanol extract thus obtained or the anthocyanin-based purple pigment of Formula 1 isolated therefrom inhibits acid oxidase and reduces uric acid in the blood, thus exhibiting anti-inflammatory action without side effects and expressing the expression of gelatinase B, which is frequently expressed in patients with gouty arthritis. By inhibiting it can be prepared a composition for preventing and treating gout containing it as an active ingredient.

Meanwhile, the composition for preventing and treating gout comprising the purple sweet potato ethanol extract or the anthocyanin-based purple pigment of Formula 1 as an active ingredient is administered orally or parenterally during clinical administration, for example, intravenous, subcutaneous, intraperitoneal or topical. It can be applied in the form of general medicine and health food.

The compositions may be formulated for oral administration such as tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs. Is formulated. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for formulation into tablets and capsules; Excipients such as dicalcium phosphate; Disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. Capsules contain liquid carriers, such as fatty oils, in addition to the substances mentioned above. In addition, the composition of the present invention can be administered parenterally, parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method. To formulate into a parenteral formulation, the composition is prepared in solution by mixing in water with stabilizers or buffers and formulated into unit dosage forms of ampoules or vials.

The effective dose of the purple sweet potato ethanol extract or the compound of Formula 1 may vary depending on the age, physical condition, weight, etc. of the patient, but is generally administered within the range of 0.1 to 30 mg / kg (weight) / day. In addition, within a daily effective dosage range is divided into once a day or several times a day.

In addition, the present invention includes a purple sweet potato ethanol extract or a health food containing an anthocyanin-based purple pigment represented by the formula (1) as an active ingredient.

The health food is a food prepared by adding the purple sweet potato ethanol extract or the compound represented by the formula (1) to food materials such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, etc. If it means to have a specific effect on health, but unlike the general medicine has the advantage that there is no side effect that can occur when the long-term use of the drug as a raw material.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Reference Example 1 Extraction of Purple Pigment from Purple Sweet Potato

Separation of the pigment from the purple sweet potato was added for about 10 times the volume of 20% ethanol solution compared to the raw purple sweet potato sample cut to 2-3 mm thickness and then left at room temperature and extracted for 8 hours. The extract was filtered through a membrane filter, followed by complete removal of ethanol at 40 ° C. with a vacuum evaporator, and then anthocyanin pigments adsorbed onto a column filled with Diion HP-20 resin. The dye adsorbed on HP-20 resin was eluted with 100% ethanol, and the solution was concentrated under reduced pressure with a vacuum evaporator to dryness to obtain an anthocyanin-based violet pigment. The obtained purple pigment is a compound of the formula (1), J. Food. Sci Nutr. Vol. 2, No. 2, p. 83-88 (1997).

[Formula 1]

이고, R₂는

,In Formula 1, R ₁

R ₂ is

,

또는

이다.

or

to be.

Example 1 Inhibition of Xanthine Oxidase by Purple Pigment

An enzyme substrate reaction experiment was carried out using xanthine as a substrate and xanthine oxidase as an enzyme.

50 mM phosphate buffer solution (pH 7.5), sample (purple pigment) and xanthine oxidase (0.5 unit / ml) were mixed and reacted at 25 ° C. for 15 minutes to allow enzyme and substrate to bind, followed by 15 μM of xanthine After the reaction was carried out at 25 ℃ 30 minutes, 1N hydrochloric acid solution was added to stop the reaction and the absorbance was measured at 290 nm. The xanthine oxidase inhibitory activity can be calculated as 50% inhibition value ((1-sampled enzyme activity / unsampled enzyme activity)) × 100. The concentration of the purple pigment that inhibits xanthine oxidase by 50% was 15.3 μΜ.

Example 2 Inhibition by Purple Pigment of Gelatinase B Expression Induced by Urea Salt

To investigate the effect of purple pigment on the amount of gelatinase B production, the mouse macrophage line RAW264.7 was passaged in 48 well plates and treated with different concentrations. It was confirmed.

Before treating the purple pigment of the present invention was changed from 10% serum medium to 1% serum medium, and then treated with 2 mg / ml monosodium urate, and cultured for 17 hours after 3 hours of sample treatment. After incubation, the shape and cytotoxicity of the cells were examined, and only the culture medium was taken and mixed with the filling buffer containing glycerol and the coloring reagent, respectively, and electrophoresed, and MMP9 expression inhibitory activity was compared with the control. In the control group, monosodium urate was used as an inducer to activate the expression of MMP9 in macrophages. Visually confirmed that the induction was divided into treatment and non-treatment groups, and the expression pattern of gelatinase B was observed in the culture medium containing the sample. It was confirmed by -PAGE. The results are shown in Figure 1, the treatment of gelatinase B expression inducing agent monosodium urate in macrophages significantly increased the expression of gelatinase B after 24 hours, but simultaneously treated with monosodium urate and purple pigment It was confirmed that gelatinase B expression was inhibited by 50% at 0.3 μg / ml in proportion to the treatment concentration of purple pigment.

Example 3: Experimental Animals and Treatments

     As a test animal, a 7 week old db / db mouse was used. In addition, seven animals in each group were used for experiments by performing group separation by a random method according to the weight range. Breeding environment is 21 ± 2 ℃, humidity 55 ± 5%, ventilation number 15-17 times / Hour, illuminance 150-300 Lux, and lighting 12 hours contrast (lighting up: 06:00, lights out: 18:00) Adjustments were made to keep SPF (specific pathogen free) constant throughout the experiment. Solid feed (Harlan, USA) and water (Autoclaved water) were fed freely and watered and fasted 12 hours before autopsy and sampling. Seven males were reported per group, and purple pigments were added orally to the Harlan feed at 0.1%. The control group received only Harlan feed.

Example 4 Measurement of Blood Uric Acid Lowering Effect of Purple Pigment by Animal Experiment

As a result of the animal experiment by the method of Example 3, the purple pigment separated from the purple sweet potato appeared to inhibit the concentration of uric acid related to gout by 85% compared to the control group (FIG. 2), thereby preventing and treating gout. It may be usefully used as a material for functional foods and pharmaceuticals.

Example 5: Toxicity Test

The anthocyanin-based purple pigment represented by Formula 1 was dissolved in dimethyl sulfoxide (DMSO), diluted with water, and then administered to the mice (10 mice per group) at 100 mg / kg, respectively, and observed for 7 days. There was no.

Formulation Example 1 Preparation of Syrup

A syrup containing purple sweet potato ethanol extract or a compound of formula 1 as an active ingredient 2% (weight / volume) according to the present invention was prepared by the following method.

Acid addition salt, saccharin and sugar of the purple sweet potato ethanol extract or the compound represented by the formula (1) were dissolved in 80 g of warm water. After the solution was cooled, a solution consisting of glycerin, saccharin, spices, ethanol, sorbic acid and distilled water was prepared and mixed thereto. Water was added to this mixture to 100 ml. The addition salt can be replaced with other salts according to the examples.

Formulation Example 2: Preparation of Tablet

A tablet containing 15 mg of active ingredient is prepared by the following method.

250 g of purple sweet potato ethanol extract or the compound of formula 1 according to the present invention was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

Formulation Example 3 Preparation of Capsule

A capsule was prepared by filling a hard gelatin capsule with 500 mg of purple sweet potato ethanol extract or the compound of Formula 1 according to the present invention.

Formulation Example 4 Preparation of Injection

200 mg of purple sweet potato ethanol extract or the compound of formula 1 according to the present invention was dissolved in 200 mg of physiological saline containing polyoxyethylene hydrogenated castro oil to prepare an injection containing a mixed extract at a concentration of 0.1.

Formulation example  5: beverage manufacturing

After dissolving the purple sweet potato ethanol extract according to the present invention or 500 mg of the compound of the formula (1) in an appropriate amount of water, vitamin C as an auxiliary component, citric acid, sodium citrate, oligosaccharides as an auxiliary agent is added, and an appropriate amount of sodium benzoate as a preservative After the addition, water was added to make 100 ml of the total amount to prepare a beverage composition. At this time, taurine, myo-inositol, folic acid, pantothenic acid, etc. may be added alone or together.

As can be seen from the above description, the purple sweet potato ethanol extract or the anthocyanin-based purple pigment of the formula 1 according to the present invention is expected to be very useful for preventing and treating gout due to its excellent hypoglycemic effect.

Claims (4)

delete Gout prevention and treatment composition comprising an anthocyanin-based purple pigment represented by the following formula (1) as an active ingredient; [Formula 1]

In Formula 1, R ₁

R ₂ is

,

or

to be. delete Gout prevention and improvement health food, characterized in that containing an anthocyanin-based purple pigment represented by the formula (1) as an active ingredient; [Formula 1]

In Formula 1, R ₁

R ₂ is

,

or

to be.

Images