KR101397044B1 - Novel Use of Asterubine - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of type 2 diabetes comprising an asterubine or a pharmaceutically acceptable salt thereof as an active ingredient, and a food composition for the prevention or improvement of type 2 diabetes.
The astubin of the present invention has an effect of increasing phosphorylation of AMPK (AMP-activated Protein Kinase) and ACC (Acetyl-CoA Carboxylase) and increasing glucose absorption capacity of muscle cells. Based on the above effect, A pharmaceutical composition for preventing or treating diabetes, and a health functional food.

Description

Novel Use of Asterubine < RTI ID = 0.0 >

The present invention relates to a novel use of Asterubine, and more particularly to the use of Astellin for the prevention or treatment of type 2 diabetes.

Metabolic syndrome (MetS) is characterized by high levels of blood fat, hypertension, insulin resistance, and central obesity (excessive fat tissue in the abdomen), and all adult diseases such as obesity, hypertension, hyperlipidemia, hyperglycemia And it becomes the basis of. The metabolic syndrome is a simultaneous manifestation of dangerous adult diseases such as arteriosclerosis, hypertension, obesity, diabetes, and hyperlipidemia in one person. Metabolic syndrome patients are living with the risk of sudden death due to cardiovascular disease as well as chronic diseases such as diabetes and hypertension. Chronic metabolic diseases caused by elevated serum cholesterol and triglycerides are the root causes of chronic metabolic diseases such as obesity, diabetes and metabolic syndrome.

The metabolic syndrome is characterized by an increase in the incidence and prevalence due to the aging of the population and changes in the environment, the mortality rate is high, and the incidence of complications is very high, resulting in enormous economic and social losses. However, most of the drugs currently used are low in efficacy It is urgent to develop therapeutic drugs in terms of improving the quality of life because of the high incidence of adverse events during long - term use. In addition, there is a decrease in the demand for therapeutic drugs in the current situation where the obesity population is rapidly increasing. Considering the complaints about side effects and efficacy against the existing drugs, it is necessary to develop a new mechanism of obesity treatment to be.

AMP-activated protein kinase (hereinafter "AMPK") has recently been shown to be involved in carbohydrate and lipid metabolism and regulates glucose transport, and has attracted attention as a new target for the treatment of type 2 diabetes (Therapie, 62, 293 -310, 2007). AMPK is a phosphorylation enzyme whose activity is regulated in response to the energy state (AMP / ATP ratio) of the cell, which is changed by the nutritional status, movement and stress of the cell (Trends Mol Med., 14, 539-549, 2008; Cell Metab., 1, 15-25, 2005). AMPK is known to regulate the phosphorylation of various enzymes involved in energy metabolism in cells, thereby affecting various physiological activities such as glucose transport, fatty acid and cholesterol synthesis (Am J Physiol Endocrinol Metab., 292, 1308- 1317, 2007). In particular, recent studies have shown that AMPK in muscle tissue and cardiac muscle promotes muscle contraction and promotes glucose uptake by transferring glucose transporter 4 (hereinafter, 'GLUT4') to the plasma membrane, regardless of insulin action, (J. Appl. Physiol., 91, 1073-1083, 2001). This suggests that a substance that activates AMPK may have the effect of accelerating absorption and consumption of glucose in muscle cells, such as exercise without exercise. Metformin, which is widely used for the treatment of diabetes, is known to lower blood glucose by inducing AMPK activation and thereby inhibiting phosphorylation / activity of ACC2 and fatty acid oxidation by affecting AMPK signaling system (J Clin Invest., 108, 1167-1174, 2001).

Therefore, the discovery of a drug that activates AMPK may lead to the development of a new concept of therapeutic agent that can mimic the improvement effect of metabolic syndrome including type 2 diabetes through exercise. However, it is not easy to develop drugs that have both safety and efficacy. Therefore, an attempt to find an ingredient that meets the type 2 diabetes treatment strategy from a safety-proven natural product and use it as a drug may be a more efficient approach than developing a therapeutic agent from a synthetic agent.

Accordingly, the inventors of the present invention have conducted research to search for a natural substance having an excellent AMPK activation effect. As a result, asterubin isolated from the hydrothermal extract of Asterias amurensis has an excellent AMPK activation effect And can be used for the prevention and treatment of type 2 diabetes, thus completing the present invention.

Accordingly, the present invention provides a method for the prevention and treatment of type 2 diabetes mellitus of asteriobin.

In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition for the prevention or treatment of type 2 diabetes comprising an asterubine having a structure represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient .

[Chemical Formula 1]

The present invention also provides a food composition for preventing or ameliorating type 2 diabetes comprising an asterubine having the structure of the following formula (1) as an active ingredient.

[Chemical Formula 1]

The astubin of the present invention has an effect of increasing phosphorylation of AMPK (AMP-activated Protein Kinase) and ACC (Acetyl-CoA Carboxylase) and increasing glucose absorption capacity of muscle cells. Based on the above effect, A pharmaceutical composition for preventing or treating diabetes, and a health functional food.

Figure 1 is an HPLC chromatogram of Amur starfish hydrothermal extract showing astutenis and asturin rich fractions.
Figure 2 shows the fractionation and MPLC chromatograms of astellin from the Amur Starfish methanol extract.
Figure 3 shows the ¹ H NMR analysis of the fractionated asbestos.
4 shows the results of ¹³ C-NMR analysis of the fractionated asbestos.
FIG. 5 shows the results of MS data analysis of the fractionated asbestos.
FIG. 6 shows the results of examining the AMPK and ACC enzyme phosphorylation effects of astellin on muscle cells (A) and the glucose uptake enhancing effect (B).
FIG. 7 shows experimental results on weight change (A) and dietary intake change (B) of mice orally administered with astrobe.
FIG. 8 shows experimental results on blood sugar change (A) and blood insulin change (B) in mice orally administered with astrobe.

The present invention provides a pharmaceutical composition for the prevention or treatment of type 2 diabetes comprising an asterubine having a structure represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 1]

In one embodiment of the present invention, the astellin having the structure of formula (I) can be used in the form of a pharmaceutically acceptable salt, wherein the acid addition salt formed by a pharmaceutically acceptable free acid useful. As the free acid, inorganic acid and organic acid can be used. As the inorganic acid, hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid and the like can be used. As the organic acid, citric acid, acetic acid, maleic acid, fumaric acid, , Acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid and arpartic acid. Preferably, hydrochloric acid is used as the inorganic acid, and methanesulfonic acid is used as the organic acid.

In addition, the astellin having the structure of the formula (1) of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by a conventional method.

In another embodiment of the present invention, the asterin having the structure of formula (1) may be fractionated from a hydrothermal extract or a methanol extract of Asterias amurensis.

More specifically, the astellin can be fractionated by injecting a hot water extract of Amur starfish into HPLC (high performance liquid chromatography), and using a methanol and water as a mobile phase to give a concentration gradient.

However, it is apparent to those skilled in the art that the astubin according to the present invention can be obtained not only from the above-mentioned Amur starfish extract but also by a general chemical synthesis method.

The present invention relates to a method for preventing or treating type 2 diabetes through the mechanism of increasing the phosphorylation of AMPK (AMP-activated Protein Kinase) and ACC (Acetyl-CoA Carboxylase) Lt; / RTI >

Accordingly, the present invention relates to a pharmaceutical composition for the prevention or treatment of type 2 diabetes mellitus, comprising asterubine or a pharmaceutically acceptable salt thereof as an active ingredient, the use of astelbin for the manufacture of a therapeutic agent for type 2 diabetes, And administering to the subject a therapeutically effective amount of astelin or a pharmaceutically acceptable salt thereof. The present invention also provides a method of treating type 2 diabetes.

In another embodiment of the present invention, the pharmaceutical composition for prevention or treatment of type 2 diabetes comprising the above-mentioned asterubine or a pharmaceutically acceptable salt thereof as an active ingredient is characterized in that, for 100 parts by weight of the pharmaceutical composition, Or a pharmaceutically acceptable salt thereof may be contained in an amount of 0.01 to 90 parts by weight, 0.1 to 90 parts by weight, 1 to 90 parts by weight, or 10 to 90 parts by weight, but not limited thereto, And the degree of progress.

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In another embodiment of the present invention, the pharmaceutical composition for the prevention or treatment of type 2 diabetes comprising the astellin or a pharmaceutically acceptable salt thereof as an active ingredient may be a carrier, an excipient, a disintegrant, a sweetener, One or more adjuvants selected from the group consisting of a lubricant, a lubricant, a lubricant, a flavoring agent, an antioxidant, a buffer, a bacteriostatic agent, a diluent, a dispersant, a surfactant, a binder and a lubricant.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

In another embodiment of the present invention, the pharmaceutical composition for preventing or treating type 2 diabetes comprising astellin or a pharmaceutically acceptable salt thereof as an active ingredient may be in the form of granules, powders, coated tablets, tablets, pills , Capsules, suppositories, gels, syrups, juices, suspensions, emulsions, drops, or liquids.

According to one embodiment of the present invention, the pharmaceutical composition may be administered orally, intraarterally, intraperitoneally, intramuscularly, intraarterally, intraperitoneally, intrasternally, transdermally, nasally, inhaled, topically, rectally, ≪ / RTI > can be administered to the subject in a conventional manner.

The preferred dosage of the above astrubin may vary depending on the condition and body weight of the patient, the kind and degree of disease, the drug form, the administration route and the period, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, the daily dose may be 0.01 to 1,000 mg / kg, specifically 0.1 to 1,000 mg / kg, more specifically 0.1 to 100 mg / kg, though it is not limited thereto. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.

The present invention also provides a food composition for preventing or ameliorating type 2 diabetes comprising an asterubine having the structure of the following formula (1) as an active ingredient.

[Chemical Formula 1]

The above food composition has a preventive or ameliorative effect on type 2 diabetes and can be effectively used for the production of health functional food and general food which can exhibit an antidiabetic effect.

In another embodiment of the present invention, the food composition comprising the astellin as an active ingredient comprises 0.01 to 90 parts by weight, 0.1 to 90 parts by weight, 1 to 90 parts by weight of astellin per 100 parts by weight of the total food composition, Or 10 to 90 parts by weight, although the present invention is not limited thereto.

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In another embodiment of the present invention, the food composition may further comprise at least one additive selected from the group consisting of organic acids, phosphates, antioxidants, lactose casein, dextrin, glucose, sugar and sorbitol. The organic acid can be, but is not limited to, citric acid, fumaric acid, adipic acid, lactic acid or malic acid, and the phosphate can be sodium phosphate, potassium phosphate, acid pyrophosphate or polyphosphate (polymeric phosphate) But are not limited to, natural antioxidants such as polyphenols, catechins, alpha-tocopherol, rosemary extract, licorice extract, chitosan, tannic acid or phytic acid.

In still another embodiment of the present invention, the food composition containing the astellin as an active ingredient may contain various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants And a carbonating agent for use in carbonating beverages such as a thickening agent (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, can do. In addition, the food composition according to one embodiment of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. The proportion of such an additive is generally, but not limited to, 0 to 20 parts by weight, 5 to 20 parts by weight, or 5 to 15 parts by weight per 100 parts by weight of the composition of the present invention.

According to one embodiment of the invention, the formulation of the food composition may be in the form of solid, powder, granule, tablet, capsule, liquid or drink, although not limited thereto.

In addition, the food composition containing astelin as an active ingredient is not limited to a confectionery, a sugar, an ice cream, a dairy product, a meat product, a fish meat product, a tofu or a jelly, an edible oil, a noodle, Dried products, cut products, fruit juice, vegetable juice, mixed juice, chips, noodles, livestock processing of dried fruits, vegetables, fruits or vegetables, health foods, seasonings, ice, ginseng products, kimchi pickles, It can be used for the production of foods such as food, fish processing food, milk processed food, fermented milk food, soybean food, cereal food, fermented microorganism food, confectionery bakery, condiments, meat processing industry, acidic beverage, licorice and herb.

According to one embodiment of the present invention, when the food composition containing astellin as an active ingredient is used for the production of a health functional food, 0.01 to 15% by weight or 0.1 to 10% by weight of the total food can be used And when used for the production of beverages, 0.02 to 10% by weight or 0.3 to 1% by weight of the total beverage can be used.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> From the Amur Starfish Asturban's  Fraction

One. Heat number  Fractions from the extract

500 μl of Amur starfish hydrothermal extract (100 mg / ml) was injected into HPLC [C ₁₈ column (21.5 × 250 mm)] and concentration gradient solvent condition (MeOH 5% → 62%, 40 minutes) mg, 0.2%) and astrubin rich fraction. After fractionation, NMR and MS data analysis showed that the structure of the compound was Astellin.

The content of ascorbic acid in the aqueous starch extract of Amur starfish was found to be about 0.2%, and the HPLC chromatogram showing the astutenin and ascorbic acid rich fraction is shown in FIG.

2. Fraction from methanol extract

Amur starfish 6.5 kg (wet weight) was extracted twice with MeOH at room temperature for 2 weeks to obtain 241 g of MeOH extract. After that, the extract was suspended in water, and the solvent fractions were sequentially fractionated using n- hexane, EtOAc, and n- BuOH to prepare respective solvent fractions. A total of 12 fractions were obtained using a silica gel VLC: MC-MeOH gradient solvent. 8 fractions (4.4 g) were subjected to C ₁₈ preparative medium pressure liquid chromatography (MPLC) [MeOH-H ₂ O], and the resulting fractions were repeatedly subjected to C ₁₈ HPLC [C ₁₈ column (21.5 × 250 mm ), MeOH-H ₂ O] to separate the asterubine. The fractionation of astellin from the methanol extract and the MPLC chromatogram are shown in Fig.

The results of ¹ H NMR, ¹³ C-NMR and MS data analysis of the fractionated astelin are shown below and in Figures 3 to 5.

^{1 H-NMR (250 MHz,} D 2 O) δ H 2.82 (6H, s), 2.98 ( see Fig. 3) (2H, t, J = 6.45), 3.47 (2H, t, J = 6.34),

¹³ C-NMR (63 MHz, D ₂ O)? _C 37.83 (CH ₃ ) 38.11 (CH ₃ ) 49.51 (CH ₂ ) 156.16 (C)

FAB-MS m / z : 196 [M + H] &lt; ⁺ &gt;

< Example  2> Asturban's In muscle cells  AMPK phosphorylation promoting effect and In muscle cells  glucose Absorption capacity  Review of the enhancement effect

In order to measure AMPK, ACC enzyme activation and glucose uptake in L6 myotube cells against astrobe, the efficacy of the compounds as described below was measured by applying the method described in the literature. (Schlattner, et al ., J Biol Chem . 279, pp 43940-51, 2004; Smwar R et al ., Clin Ther . 20, 125-40, 1998).

As a result, as shown in FIG. 6A, it was confirmed that the concentration of AMPK and ACC increased in a concentration-dependent manner in the group treated with astrobase compared to the control group. Also, referring to FIG. 6B, it was confirmed that glucose uptake was also higher in the astubin-treated group than in the control group. This was similar to Metformin, a diabetes drug.

< Example  3> Type 2 diabetes model mice db / db mice For in vivo  Review effects

AMPK activation has been reported to increase intracellular glucose uptake and overcome insulin resistance and decreased glucose uptake as in type 2 diabetes. (Hutberca, et al., AM J Physiol 272, ppE262-E266, 1997). In addition, as a specific metabolic change according to AMPK activation, AMPK, which is a phosphorylated active form, has an effect of directly phosphorylating ACC protein producing malonyl-CoA as a key enzyme of fatty acid synthesis (Merrill G F., et al. , Am J Physiol 272, ppE1107-E1112, 1997).

Thus, based on the effect of Asterubine shown in Example 2, an in vivo experiment was conducted with db / db mice, a type 2 diabetic model rat. In the experimental group, 5-week-old rats were used for the experiment from 6 weeks of age after 1 week of adaptation period. Saline (0.9% NaCl) was orally administered to the control db / db group daily. Metformin group and Asterubine group were 250 mg / kg BW and 100 mg / kg BW, respectively. (Asan), triglyceride (Asan), and HDL cholesterol (Asan) were measured by measuring body weight, dietary intake, and blood glucose levels at 5 days during the experiment. Were measured using each ELISA kit.

The results are shown in Fig. 7, Fig. 8 and Table 1. Fig.

7, there was no difference in food intake (see FIG. 7B) compared to the control db / db when oral administration of astelin (100 mg / kg BW) was observed, but the weight was decreased by 10.7% (See FIG. 7A). In FIG. 8, the content of insulin in the blood relative to the control of blood glucose and blood glucose was 24.8% (see FIG. 8A) and 29.6% (blood insulin, see FIG. 8B) ), Respectively.

The results of analysis of various lipids in blood are shown in Table 1 below. Referring to Table 1, total cholesterol, triglyceride and LDL cholesterol were reduced by 26.0%, 50.3% and 42.5%, respectively, as compared with the db / db group .

Group Total cholesterol
(mg / dl) Triglyceride
(mg / dl) HDL cholesterol
(mg / dl) LDL cholesterol
(mg / dl) Wild type (Lean) 140.4 ± 11.7 61.3 ± 5.0 82.1 ± 4.7 46.0 + - 6.7 db / db 20.8 ± 9.2 167.7 ± 11.1 76.1 ± 7.0 111.1 ± 11.4 db / db + metformin
(250 mg / kg) 194.3 ± 14.7
(10.4% ↓) 93.7 ± 9.2
(44.2% ↓) 113.0 + - 8.6
(48.4% ↑) 62.6 ± 4.0
(43.7% ↓) db / db + Asterubine
(100 mg / kg) 163.3 ± 13.6
(26.0% ↓) 83.4 ± 1.5
(50.3% ↓) 82.7 ± 10.2
(8.6% ↑) 63.9 ± 13.0
(42.5% ↓)

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (6)

1. A pharmaceutical composition for the prevention or treatment of type 2 diabetes comprising an asterubine having the structure of the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Chemical Formula 1]

The method according to claim 1,
Wherein the astellin is fractionated from a hydrothermal extract or a methanol extract of Asterias amurensis. The method according to claim 1,
Wherein the astellin increases phosphorylation of AMP-activated Protein Kinase (ACC) and Acetyl-CoA Carboxylase (ACC) and increases glucose absorption capacity of muscle cells. delete 1. A food composition for preventing or improving type 2 diabetes comprising an asterubine having a structure of the following formula (1) as an active ingredient.
[Chemical Formula 1]

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