KR20120113320A - Composition comprising the extract of capsosiphon fulvescens for preventing or treating diabetes and diabetes complication, and antioxidant - Google Patents

Abstract

PURPOSE: A pharmaceutical composition and a health functional food composition containing Capsosiphon fulvescens extract is provided to ensure bio-affinity and low toxicity expression and to prevent and treat diabetes/diabetes complication. CONSTITUTION: A composition for preventing or treating diabetes and diabetes complication contains Capsosiphon fulvescens extract as an active ingredient. The composition includes a pharmaceutical composition or a food composition. A method for preparing the extract comprises: a step of extracting Capsosiphon fulvescens with alcohol and pulverizing; a step of extracting the powder with one or more kinds of organic solvents; and a step of a step of performing gel filtration chromatography. The organic solvent extract is a hexane fraction.

Description

Composition comprising the extract of Capsosiphon fulvescens for preventing or treating diabetes and diabetes complication, and antioxidant}

The present invention relates to an antioxidant agent containing an extract as an active ingredient, and a composition for preventing or treating diabetes mellitus and diabetic complications. More specifically, the present invention relates to an antioxidant agent containing no extract of side effects, ensuring safety, and having excellent efficacy as an active ingredient, a pharmaceutical composition for preventing, treating or improving diabetes and diabetic complications, and a health functional food composition.

The Maial reaction, also called non-enzymatic gliding or browning, was first discovered in 1912 by the French scientist Louis Camille Mailard. Working on the reaction in 1912, the scientist Mailard noticed a change in color from yellow to brown when reducing sugars were heated with amino acids at high temperatures (Maillard, 1912, Comptes Rendus Hebdomadaires Des Seances De L Academie Des Sciences). 154: 66-68).

The Mial response has previously been widely used in the food industry to control the texture of food, but more recently is known for its association with various diseases. In particular, diseases that are closely related to the MIAL response include neurodegenerative diseases such as Alzheimer's disease and diabetes and related complications (Reddy and Beyaz, 2006, Drug Discovery Today 11: 646-54). Structural alteration of glycated proteins leads to cell damage by altering various receptors present in the cell, resulting in loss of physiological activity and toxicity, (Vlassara et al., 1994, Laboratory Investigation 70: 138-151). Glycosylated proteins are formed on short and long-lived proteins such as albumin, skin collagen, immunoglobulins, eyes, kidneys, and vascular tissues (Thorpe and Baynes, 2003, Amino Acids 25: 275-281).

The gliation reaction is a non-enzymatic reaction between the carbonyl group of the reducing sugar and amino groups such as proteins, nucleic acids, and phospholipids to form the final glycated end-product (AGEs), which forms the initial Schiff base and oxidizes. , Condensation, dislocation and rearrangement result in the formation of an Amadori compound and finally an irreversible final glycosylation product (Reddy and Beyaz, 2006, Drug Discovery Today 11: 646-54).

Many studies have shown that gliding is closely related to the progression of various diseases associated with aging, including diabetes, arteriosclerosis, end stage renal failure, rheumatoid arthritis, and neurodegenerative diseases. Gliding and final glycation products mainly affect the function of long living proteins. Final glycation end products and final lipid oxidation products (ALEs) accelerate the progression of microvasculopathy and macrovasculopahty observed in diabetes. AGEs are found in the blood and tissues of people with diabetes, as well as in various pathological conditions such as atherosclerosis, Alzheimer's, end-stage renal failure, rheumatoid arthritis, and cirrhosis of the liver (Sebekova et al., 2002, J Hepatol 36:66 -71).

It is known that final glycation products have chemical modifications and cross-links that affect the structure and function of proteins, lipids and DNA tissues, resulting in decreased tissue function and diabetes and diabetic complications (Jakus and Rietbrock). , 2004, Physiol Res 53: 131-42).

Figure 1 shows the reaction process of the non-enzymatic Maillard reaction and the production of AGEs. In the initial stage of the reaction, the non-enzymatic reaction between the carbonyl group of the reducing sugar and the first amino acid (lysine arginine) group of the protein is used to shift-base. Which further forms Amadori rearrangement. Then a complex process of intermolecular oxidation, dehydrogenation and condensation produces some fluorescent, condensates, such as AGRs (Derbre et al., 2010, Analytical and Bioanalytical Chemistry 398: 1747-1758).

On the other hand, the use of a transition metal chelator or free radical scavenging ability or by breaking the cross-linking of AGEs prevents oxidation, ie an AGE inhibitor or breaker that inhibits reactive dicarbonyl species. Many studies have been conducted.

Aminoguanidine (AG), pyridoxamine, 2,3-diamino-phenazine, OPB-9195, and tenilsetam are reactive carbonyl intermediates N-phenacetylthiazolium bromide (PTB) and ALT-711 act as AGE-cross-link breakers by eliminating reactive carbonyl intermediates.

Signaling through RAGE may be inhibited by antisense oligodeoxynucleotides (AS-ODNs), RAGE antibodies or soluble RAGEs.

Aminoguanidines (AG) are well known end glycosylation inhibitors, in vitro and in vivo products such as methylglyoxal, Amadori products such as 3-deoxyglucosone, intermediates, and Reaction with the carbonyl group of the reducing sugars inhibited the formation of the final glycosylated product.

2 shows the process of inhibiting AGEs production by aminoguanidine, and illustrates the process by which protein binding by AGEs is inhibited by aminoguanidine.

On the other hand, green algae, Capsosiphon fulvescens (CF), is one of the important seaweeds for food in Korea and Japan. It has a unique flavor and soft texture, and is widely consumed as a traditional folk food in Korea. Essential and free amino acids and proteins are very high, whereas lipids, carbohydrates and dietary fiber are very low (Hwang et al., 2008, Applied Phycology 20: 147-151).

It is known to induce apoptosis of gastric cancer cells through the IGF-IR-mediated PI3K / Akt pathway using the AGS cell line, and the polysaccharides contained in the maestro stimulate the growth of IEC-6 cells by activating the MAPK signaling pathway. (Go et al., 2010, Mar Biotechnol (NY)). In addition, studies have been conducted to verify the effect of extracts of falcons on alcohol-related diseases (Hwang et al., 2008, Food and Chemical Toxicology 46: 2653-2657).

However, although Maesui has been consumed as a food for a long time, the treatment and prevention effects of diabetes / diabetic complications have not been studied or known to date.

Diabetes mellitus is one of the most common chronic adult diseases, and it is estimated that about 5% of the total population in Korea is at least 2.5 million. In advanced countries, the number of diabetic patients is increasing every year, and in Korea, the number of patients is expected to increase gradually with the improvement of living standard and westernized lifestyle. Diabetes is a metabolic disease in which the body's ability to use glucose normally is impaired and blood sugar levels increase, causing excess sugar to be excreted in the urine as blood sugar levels increase. Therefore, diabetes occurs when insulin is deficient or insulin resistance occurs. Substances that lower insulin glucose levels in the blood by replacing insulin are used as diabetes treatments. Conventional diabetes treatments are limited in their use due to various side effects. In addition, since the risk of diabetes is due to various complications caused by diabetes, prevention and treatment of complications are very important.

Therefore, there is a continuing need to develop new antioxidant agents that are equivalent to or better than the conventional antidiabetic agents and more safe for the human body, and materials for preventing, treating or improving diabetes / diabetic complications.

The present inventors have repeatedly studied to develop a material from natural products that have no side effects and ensure safety and excellent antioxidant effect, treatment and prevention effect of diabetes / diabetic complications, and as a result, a large number of habitats throughout Korea The present invention has been completed by confirming the anti-oxidant effect and the efficacy of treating, preventing and improving diabetes / diabetic complications that have not been achieved.

Accordingly, an object of the present invention is to provide a composition for preventing, treating or improving diabetes and diabetic complications, which has no toxicity and no side effects and has excellent efficacy as an active ingredient extract.

Another object of the present invention is to provide a pharmaceutical composition and a health functional food composition containing the composition.

Still another object of the present invention is to provide an antioxidant agent composition having no toxicity and no side effects, and having excellent potency extract as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

In order to achieve the above object, according to an aspect of the present invention, the present invention provides a composition for preventing or treating diabetes mellitus and diabetic complications containing the extract as an active ingredient.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus and diabetic complications comprising the above composition containing the extract as an active ingredient.

According to another aspect of the present invention, the present invention provides a health functional food composition for preventing or improving diabetes mellitus and diabetic complications comprising the composition containing the extract as an active ingredient.

According to another aspect of the present invention, the present invention provides an antioxidant agent composition containing the extract as an active ingredient.

According to another aspect of the present invention, the present invention comprises the steps of i) obtaining and powdering an alcoholic extract of perennial, ii) dissolving the powder obtained in step i) in water and then using a polar organic solvent. Extracting sequentially accordingly; And iii) a method for preparing an antioxidant preparation composition comprising the extract of the perennial extract as an active ingredient, or a method for preparing or preventing diabetic / diabetic complications, comprising performing gel filtration chromatography on the organic solvent extract obtained in step ii). to provide.

Hereinafter, the present invention will be described more specifically.

We extracted perennial using 70% ethanol and used solvent fractionation to isolate the active material of perennial for gliding. The total polyphenol content of the extracts and the hexane fractions isolated from the maeseng was confirmed the antioxidant effect and the inhibitory effect using the reducing power of iron ions to complete the present invention (Fig. 3 to 6).

Accordingly, the present invention provides a composition for preventing or treating diabetes mellitus and diabetic complications containing the extract as an active ingredient.

The present invention also provides an antioxidant agent composition containing the extract as an active ingredient.

In the present invention, the composition is preferably a pharmaceutical composition, health functional food composition or cosmetic composition, but is not limited thereto.

The composition of the present invention is characterized by inhibiting the anti-oxidative effect and the gliding to inhibit the production of the final glycation products associated with diabetes and diabetes complications.

In the present invention, the extract should include any of the extracts, fractions and purified products obtained in each step of extraction, fractionation and purification, any dilution or concentrate thereof, or dried products thereof.

In the present invention, the extraction solvent is preferably selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms and mixed solvents thereof, and the extraction is more preferably extracted using 70% ethanol as a solvent. It is not limited to this.

The alcohol extract may be further fractionated using an organic solvent having a different polarity, and may be further purified by various chromatography. Although not limited thereto, the composition is characterized in that it comprises a hexane (hexane) fraction of the perennial.

Although not limited thereto, the hexane fraction may be separated and purified using a known technique such as silica gel 60 or sepadex LH-20 column. In addition, the composition is not limited thereto, and the composition includes a fraction eluted with a mixed solution of hexane and ethyl acetate using chromatography. More preferably the composition comprises a silica gel 80:20 (hexane: ethyl acetate) fraction.

In the composition of the present invention, preferably, the perilla extract may be included in an amount of 0.0001 to 90% by weight based on the total weight of the composition.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating diabetes mellitus and diabetic complications comprising the above composition containing the extract as an active ingredient.

As discussed above, the maesengyi of the present invention and the maeyi extracts (or fractions) comprising the same inhibit the production of the final glycation product AGE, the composition of the present invention is diabetic or diabetic retinopathy resulting from the production of the final glycation products It may be useful for the prevention and treatment of diabetic complications, such as diabetic cataracts, diabetic nephropathy, diabetic neuropathy, diabetic fatty liver, and the like.

The pharmaceutical compositions of the present invention may be formulated in the form of powders, granules, tablets, capsules, injections, creams, gels, patches, sprays or ointments by conventional methods known in the art.

In addition, the pharmaceutical composition containing the extract of the present invention as an active ingredient may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Typically carriers, excipients and diluents include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose , Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose, or the like in the extract. Or lactose, gelatin, or the like is mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. 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 base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts 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. For the preferred effect, the extract of the present invention is administered at 0.001 to 2000 mg / kg on a daily basis, it is preferable to administer at 0.01 to 50 mg / kg to be more effective. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

Extracts of the invention can be administered by a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

According to another aspect of the present invention, the present invention provides a health functional food composition for preventing or improving diabetes mellitus and diabetic complications comprising the composition containing the extract as an active ingredient.

As discussed above, the maesengyi of the present invention and the maeyi extracts (or fractions) comprising the same inhibit the production of the final glycation product AGEs, the composition of the present invention is diabetic or diabetic retinopathy resulting from the production of the final glycation products It can be useful for the prevention and improvement of diabetic complications, such as diabetic cataract, diabetic nephropathy, diabetic neuropathy, diabetic fatty liver, and the like.

In the present specification, the term "food" means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. Includes food, food additives, nutraceuticals and beverages. In the present specification, the health functional food means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and the term "functionality" refers to the structure of the human body and It means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action with respect to function.

In the food composition of the present invention, the mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). The extract is not limited thereto and may be included in an amount of 0.0001 to 90% by weight based on the total weight of the composition. The food of the present invention may further comprise a food acceptable food supplement, and may be formulated in the form of pills, powders, granules, acupuncture, tablets, capsules or beverages.

For example, the health functional food in the form of tablets may be prepared as it is or by adding an excipient, a binder, a disintegrating agent or other additives to make a granular form in a suitable manner, and then compressing by adding a lubricant or the like or preparing a health in a tablet form. It is made by directly compressing the functional food as it is or by mixing it evenly with an excipient, binder, disintegrant or other suitable additives, or mixing the health functional food as it is or evenly adding the appropriate additive to the prepared granules, The excipient, binder or other suitable additives are added to the functional food, and the powder mixed evenly is wetted with a solvent, the wet powder is molded into a mold at low pressure, and then dried and prepared by a suitable method. In addition, the nutraceutical can be added to the health functional food in the form of tablets, if necessary, can be avoided with a suitable epidermis.

Among the health functional foods in the form of capsules, the hard capsules are usually prepared by mixing the capsules evenly with the health functional foods or excipients suitable for the health functional foods, granulated by a suitable method or granulated with a suitable epidermal agent as it is or Soft capsules are prepared by filling them, and soft capsules are usually filled with capsules containing glycerin or sorbitol in a capsule form containing gelatin or appropriate excipients suitable for health functional foods or health functional foods. It is molded and prepared, and a coloring agent preservative etc. can be added to the said capsule base as needed.

The health functional food in the form of a cyclic form is usually prepared by mixing excipients, binders, disintegrants and the like evenly in the health functional foods and forming them into spherical forms in a suitable manner. Or they may be greeted with a suitable substance.

In addition, the composition of the present invention can be used as a food supplement of health supplements for the prevention of related diseases. Foods to which the extract of the present invention may be added include various foods, for example, beverages, gums, teas, vitamin complexes, dietary supplements, and the like, which are pills, powders, granules, tablets, tablets, capsules or beverages. Available in form.

At this time, the amount of the extract in the food or beverage, in general, can be added to 0.0001 to 50% by weight of the total food weight of the food composition of the present invention, 0.0001 to 20% by weight based on 100ml for the health beverage composition Can be added.

Food additives other than the extracts defined in the present invention include food additives customary in the art, such as flavors, flavors, colorants, fillers, stabilizers and the like.

In the case of the beverage composition, there are no particular limitations on the liquid components contained in the indicated ratios other than the extract as essential components, and may contain various flavors or natural carbohydrates, etc. as additional components, as in general beverages. Examples of the natural carbohydrates include, for example, monosaccharides such as glucose and fructose, for example, disaccharides such as maltose and sucrose, for example, conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and xylitol. Sugar alcohols such as sorbitol and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., 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 drinks, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a cosmetic composition containing the essence of the present invention as an extract active ingredient. As described above, since the composition according to the present invention has an antioxidant effect, it can be usefully used as a cosmetic composition.

The components included in the cosmetic composition include components commonly used in cosmetic compositions in addition to the hexane fractions separated from maeseng and hexane fractions, and the silica gel 80:20 (hexanes: ethyl acetate) fractions separated from the hexane fractions, for example, antioxidants, Conventional adjuvants such as stabilizers, solvents, vitamins, pigments and flavorings, and carriers. In addition, the cosmetic composition may further include a skin absorption promoter to promote its effect.

Cosmetic compositions of the present invention can be prepared in any formulation conventionally prepared in the art, for example solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, oils, powder foundations, emulsions It may be formulated as a liquid foundation, wax foundation or spray. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

The hexane fractions and hexane fractions separated therefrom and the silica gel 80:20 (hexane: ethyl acetate) fractions separated from the hexane fractions according to the present invention may be used in an amount of 0.001 to 50% by weight relative to the total weight of the cosmetic, but are not limited thereto. It is not.

According to another aspect of the present invention, the present invention comprises the steps of i) obtaining and powdering an alcoholic extract of perennial, ii) dissolving the powder obtained in step i) in water and then using a polar organic solvent. Extracting sequentially accordingly; And iii) a method for preparing an antioxidant preparation composition comprising the extract of the perennial extract as an active ingredient, or a method for preparing or preventing diabetic / diabetic complications, comprising performing gel filtration chromatography on the organic solvent extract obtained in step ii). to provide.

Although not limited thereto, the organic solvent extraction may be performed in the order of hexane, chloroform, ethyl acetate, and butanol (see FIG. 3).

Although not limited thereto, the gel filtration chromatography may be performed using a silca gel 60 or Sepadex LH-20 column.

Although not limited thereto, the organic solvent extract in step iii) is preferably a hexane fraction. Hexane fractions were shown to have excellent antioxidant and antiglyzing effects (see FIGS. 4 and 5).

Although not limited thereto, the eluting of the gel filtration chromatography in step iii) is performed using a mixed solution of hexane and ethyl acetate, and the composition preferably comprises a silica gel 80:20 (hexane: ethyl acetate) fraction. Do. The antigelling effect of the silica gel 80:20 (hexane: ethyl acetate) fraction of the hexane layer was found to be excellent (see FIG. 6).

As described above, the maesengyi extract according to the present invention is effective in preventing or treating diabetes and diabetic complications because it has a gliding inhibitory effect. In addition, the maesengyi yi extract according to the present invention as a natural product extract has a very low side effects and toxic expression rate and can promote the effect of preventing or treating diabetes / diabetic complications biocompatible.

In addition, the maesengyi extract according to the present invention has a high total polyphenol content and has an antioxidant effect, so can be effectively used as an antioxidant agent.

In addition, the present invention is expected to increase the value of the perennial by increasing the application range of the extract from simple food to prophylactic or therapeutic drugs and to increase the income of workers in related industries.

Figure 1 shows the reaction of the non-enzymatic Maillard reaction and the production of AGEs.
Figure 2 shows the process of inhibiting the production of AGEs by aminoguanidine.
Figure 3 is a schematic of the extraction, separation and purification steps to obtain a large fraction of the antioxidant and antiglycation effect of the larvae.
Figure 4 shows the results of evaluating the antioxidant capacity using the reducing power of the total polyphenols and iron ions of the fractions with different polarity and life. Each value was expressed as the average ± standard deviation of the results of three replicates. The iron ion reducing power (FRAP) was expressed as mole FeSO ₄ / μg sample, and the total polyphenols were expressed as μg-gallic acid equivalents / mg of the extract.
Figure 5 shows the results of evaluating the anti-gliding ability of the fractions of different life and polarity. Antiguan activity of quantitative control aminoguanidine (10 mM) and 70% ethanol extract (CFE) of C. falciparum and CFE-H, CFE-C, CFE-E, CFE-B and CFE-D.
Figure 6 shows the results of evaluating the anti-glitability of the silica gel column fractions of the hexane layer of the perennial active fraction. The percentage of gliding inhibition activity was calculated in comparison to the blank group (100%), the value was mean (n = 3) ± SD, and the final concentration of aminoguanidine was 10 mM. P <0.05

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1 Extraction, Separation, and Purification to Obtain Fractions with High Antioxidant and Antiglycation Effects

Figure 3 is a schematic of the extraction, separation, and purification steps to obtain a fraction of the anti-oxidant and anti-gliding effect of the larvae. After washing with water three times or more, 500 g of Capsosiphon fulvescens dried using a lyophilizer was obtained by reflux cooling for 3 hours using 5% of 70% ethanol, followed by centrifugation at 7,500 rpm, 30 min, and 4 ° C. Got. The supernatant was again used to remove insoluble matter that may remain in the centrifuged supernatant. 42 filtered.

The filtered supernatant was distilled under reduced pressure and then lyophilized to obtain 70% ethanol extract (CFE) in powder form. The CFE thus obtained was dissolved in water, and then the polarity of the organic solvent was sequentially changed. -D) The primary fractions were separated according to polarity as fractions. After adsorbing CFE-H, which has the highest antioxidant and anti-glycolytic activity, on silica gel chromatography, 10 fractions were obtained by varying the concentration of <hexane: ethyl acetate> mixed eluent. The highest <80% hexane: 20% ethyl acetate> fraction (CFE-H-2) was separated again using Sephadex LH-20 column (2 × 35 cm).

Example 2 Antioxidative Effect of Total Lifespan and Fractions Separated therefrom (Total Polyphenol, Iron Ion Reducing Power)

Antioxidant activity was measured by total polyphenol content and iron ion reducing power. Total polyphenol content was measured using Folin-Ciocalteau reagent (Anesini et al., 2008, Agricultural and Food Chemistry 56: 9225-9229). 0.1 ml of sample, 2 ml of methanol, and 0.2 ml of 2 N Folin-Ciocalteau reagent were added to each tube and reacted for 3 minutes. After adding 1 ml of 15% Na ₂ CO ₃ and reacting at room temperature for 2 hours, the absorbance was measured at 765 nm using Gallic acid as a standard. The iron ion reducing power was measured for 5 minutes with the sample diluted in each concentration of FRAP test solution dissolved in 0.3 M acetate buffer in 10 mM 2, 4, 6-tripyridyl-s-triazine (TPTZ) and 20 mM FeCl ₃ -H ₂ 0. The reaction was measured for absorbance at 593 nm.

As shown in Figure 4, the result of measuring the total polyphenol content CFE showed a polyphenol content capable of exhibiting a large amount of antioxidant capacity with a 18.61 ㎍ gallic acid / mg sample, and again among the fractions fractionated by organic solvent The CFE-H layer (hexane fraction) and the CFE-E (ethyl acetate fraction) layer showed high antioxidant indices of 24.25 and 24.53 μg gallic acid / mg samples. In iron ion reducing power (FRAP), CFE was 71.42 mole FeSO ₄ / mg sample, which showed high iron ion reducing power, that is, antioxidant effect. Among the fractions, CFE-H (hexane layer) layer was 73.34 mole FeSO ₄ / Mg sample showed excellent antioxidant capacity. Based on this, the maeseng and fractions fractionated from it can be seen that the antioxidant capacity is higher than any known antioxidants, and excellent antioxidants, and based on this, it has anti-oxidative and diabetic / diabetic complications.

Example 3 Anti-Gliding Effect of Maizei and Fractions Separated from It

Antiglycation was incubated at 37 ° C after mixing BSA 10 mg, 10 mM glyceraldahyde 1.0 ml, 1 mM DTPA and 0.02% sodium azide dissolved in 0.1 M phosphate buffer (pH 7.4). 1 ml of this mixture was transferred to 1.5 ml ep-tube (Corning Costar Co.), sealed, and reacted for 7 days in an O ₂ incubator. 100 μl of each was divided into 3 wells and transferred to a 96well plate, and the fluorescence was measured at an excitation wavelength of 370 nm and an emission wavelength of 440 nm using a spectrofluorometer (VICTOR3 ™, PerkinElmer, USA). Glycation inhibitor aminoguanidine was dissolved in distilled water.

Of CFE, which is a perennial extract, CFE-H (hexane layer), CFE-C (chloroform layer), CFH-E (ethyl acetate layer), CFE-B (butanol layer), and CFE-D (water layer) fractionated therefrom. Antiglycolytic activity experiments were conducted.

Negative control group was considered to have 100% inhibitory effect because no sugar was added. In the results of FIG. 5, CFE inhibitory effects of 100, 500 and 1000 ㎍ / ml concentration were 16.48, 51.26, 66.34% and IC ₅₀ value, respectively. Is 635.37 μg / ml. Through this, it was confirmed that the maesenggi has anti-gliding activity, and the CFE-H IC ₅₀ 571.38 ㎍ / ㎖ was the highest anti-gliding effect in the other fractionation layer in the re-fractionated layer. Based on the result of the fraction having an inhibitory effect on the formation of glycerol, it is necessary to separate and purify the active material using a silica gel column.

Example 4 Anti-Gliding Effect of Silica Gel Column Fractions of the Hexane Layer with the Largest Anti-Gliding Activity

CFE-H (hexane layer) was separated and purified by stepwise gradient CFE-H using silica gel open column, which is the best glysis inhibitory effect among the fractions. The solvent was separated into 10 aerobic fractions using ethyl acetate and hexane, and the anti-gliding activity experiments were performed for each fraction.

As shown in FIG. 6, the result showed the highest antiglycolytic activity in the hexane and ethyl acetate 80:20 (v / v) fractions (CFE-H-2). In addition, the fraction showed more than 70% of the anti-glare activity at 500 μg / ml concentration. Therefore, it was estimated that CFE-H-2 had an active substance and separated and purified by Sephadex LH-20 column chromatography.

Formulation Example 1 Preparation of Tablet

100 mg of Maeseng Yiyi extract, corn starch 90.0 mg, lactose 175.0 mg, 15.0 mg of L-hydroxypropyl cellulose, 905.0 mg of polyvinylpyrrolidone and an appropriate amount of ethanol were granulated by wet granulation and magnesium stearate 1.8 MG was added and mixed, and then one tablet was compressed to 400 mg.

Formulation Example 2 Preparation of Health Functional Food

500 mg of falcon extract

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

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

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

24.8 mg of magnesium chloride

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health functional food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above components may be mixed according to a general health functional food manufacturing method. The granules can then be prepared and used to prepare the nutraceutical composition according to conventional methods.

Although specific portions of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific techniques are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. It is therefore intended that the scope of the present invention be defined by the appended claims and their equivalents.

Claims (9)

A composition for the prevention or treatment of diabetes mellitus and diabetic complications containing the extract as an active ingredient. The composition for preventing or treating diabetes mellitus and diabetic complications of claim 1, wherein the composition inhibits gliding. A pharmaceutical composition for preventing or treating diabetes mellitus and diabetic complications comprising the composition according to claim 1. A health functional food composition for preventing or improving diabetes and diabetic complications, comprising the composition according to claim 1. Antioxidant composition containing the extract as an active ingredient. The method of claim 5, wherein the composition is an antioxidant agent composition, characterized in that the cosmetic composition. i) obtaining and powdering an alcoholic extract of perennial;
ii) dissolving the powder obtained in step i) in water and sequentially extracting the powder obtained by polarity using at least one organic solvent; And
iii) a method for preparing an antioxidant agent composition comprising a medicinal herb extract as an active ingredient comprising performing filtration by gel filtration of the organic solvent extract obtained in step ii) or a composition for preventing or treating diabetes and diabetic complications. 8. The method of claim 7, wherein the organic solvent extract in step iii) is a hexane fraction, the method of producing an antioxidant agent composition, or a composition for preventing or treating diabetes and diabetic complications, containing the extract as an active ingredient. The method of claim 8, wherein the eluting of the gel filtration chromatography in step iii) is carried out using a mixed solution of hexane and ethyl acetate, the antioxidant agent composition containing the extract of the essence as an active ingredient, or diabetes mellitus and diabetes mellitus Method for preparing a composition for preventing or treating complications.

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