KR20220073219A - A composition for improving, preventing and treating of obesity comprising Centella asiatica fermentation extract - Google Patents

Abstract

The present invention relates to a composition for improving, preventing or treating obesity and diabetes, and more particularly, by including a fermented extract of Centella asiatica as an active ingredient, improving, preventing diseases that can be caused by obesity and diabetes Or because it can be treated, it can be used as a food composition, furthermore, a health functional food or a pharmaceutical composition.

Description

A composition for improving, preventing or treating obesity and diabetes, comprising a fermented extract of Centella asiatica comprising Centella asiatica fermentation extract

The present invention relates to a composition for improving, preventing, or treating obesity and diabetes, comprising a fermented Centella asiatica extract fermented with probiotics as an active ingredient.

Obesity is the cause of various lethal metabolic diseases such as cardiovascular disease, high blood pressure and Type II diabetes. Adipocytes not only store lipids as a long-term energy source but also produce various adipokines involved in the metabolism and inflammation of adipocytes and non-adipocytes. It causes chronic inflammation and related metabolic diseases such as insulin resistance.

In recent years, the incidence of metabolic syndrome, also known as Type II diabetes, is increasing worldwide due to obesity due to lack of exercise along with a high-calorie diet. It is predicted to do Obesity caused by abnormal local fat accumulation, such as abdominal obesity, provides various causes that can seriously induce arterial atheropathy, such as high blood pressure, hyperlipidemia, blood clotting disorder, vascular inflammation, and insulin resistance with abnormal increase in insulin secretion. Ultimately, it becomes a factor in increasing the prevalence of fatal cardiovascular disease.

Excessive intake of fat causes the accumulation of triglyceride (TG) in various tissues, accompanied by an increase in lipolysis and an increase in circulating fatty acids in the blood, and insulin resistance in adipocytes, resulting in muscle and pancreas. and fat accumulation in non-adipocytes such as the liver. When insulin resistance is induced in adipocytes, excess fatty acid binding and transport proteins lead to increased fatty acid uptake by non-adipocytes, particularly in the case of muscle cells, which adversely affects insulin-mediated glucose metabolism, while at the same time, the pancreas converts free fatty acids into free fatty acids. Long-term exposure repeats a vicious cycle in which insulin secretion is impaired by a so-called lipotoxicity mechanism. In this case, a high concentration of free fatty acids is also accumulated in the liver, resulting in resistance to insulin, and a large amount of glucose begins to be liberated from the liver. Accumulation of TG in hepatocytes causes non-alcoholic fatty liver disease (NAFLD), and causes fat accumulation in hepatocytes mainly responsible for glucose metabolism, resulting in secondary steatohepatitis, and finally It causes fibrosis by necrosis of hepatocytes. Therefore, the balance of hepatocyte lipid synthesis and decomposition has been an important therapeutic target that can suppress the induction of insulin resistance and NAFLD caused during metabolic syndrome.

Various metabolic syndrome treatments are currently being developed, and in recent years, proper control of oxidative stress, which has been pointed out as a major cause of diabetes and related complications, along with proper blood sugar control, has emerged as the most important method for diabetes treatment, so side effects are low and The development of more effective α-glucosidase blockers or antioxidants is being attempted.

The treatment of obesity includes drug therapy, etc. Drug therapy is a treatment method in which an anti-obesity agent is administered to an obese patient. Obesity drugs include appetite suppressants, metabolic suppressants, absorption inhibitors, and thermogenesis promoters, but problems such as lethargy and personality changes occur when consumed for a long time.

Therefore, there is a need for a composition for improving, preventing or treating obesity and diabetes using a natural material that does not cause the above problems.

Republic of Korea Patent No. 2132066 Republic of Korea Patent No. 1915071

An object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity and diabetes containing a ferment extract of Centella asiatica as an active ingredient.

In addition, another object of the present invention is to provide a food composition for preventing or improving obesity and diabetes containing a fermented extract of Centella asiatica as an active ingredient.

The pharmaceutical composition capable of preventing or treating obesity and diabetes of the present invention for achieving the above object may contain a fermented extract of Centella asiatica as an active ingredient.

The fermented Centella asiatica extract may be a fermented extract obtained by fermenting the Centella asiatica solvent extract as a strain.

The strain is Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus sakei ( Lactobacillus sakei ), Leuconostoc mesenteroides ( Leuconostoc mesenteroides ) and Lactobacillus buchneri ( Lactobacillus buchneri ) at least one selected from the group consisting of have.

The centella asiatica solvent extract may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The centella asiatica solvent extract may be processed for 5 to 60 minutes with an ultrasonicator of a frequency of 20 to 50 kHz and a power of 50 to 700 W.

The extraction temperature during the ultrasonic treatment may be 50 to 90 ℃.

In addition, the food composition capable of preventing or improving obesity and diabetes of the present invention for achieving the above other object may contain a fermented extract of Centella asiatica as an active ingredient.

Since the composition for improving, preventing or treating obesity and diabetes comprising the fermented Centella asiatica extract of the present invention as an active ingredient can improve, prevent or treat diseases that may be caused by obesity and diabetes, a food composition, furthermore, a health functional food Or it may be utilized as a pharmaceutical composition.

1 is a diagram showing the expression of glucose transporter 4 (GLUT4) in a fermented extract of Centella asiatica prepared according to Example 1 of the present invention.
2 is a view showing the expression of PPAR-α in the fermented Centella asiatica prepared according to Example 1 of the present invention.

The present invention relates to a composition for improving, preventing, or treating obesity and diabetes, comprising a fermented Centella asiatica extract fermented with probiotics as an active ingredient.

Hereinafter, the present invention will be described in detail.

The composition capable of improving, preventing or treating obesity and diabetes of the present invention contains a fermented Centella asiatica extract as an active ingredient.

The centella asiatica ( Centella asiatica ) is a perennial creeping herb, belonging to the Umbelliferae family, and is a plant that grows mainly in hot and humid places, and inhabits wetlands such as Africa, India, and Sri Lanka. In Korea, it mainly lives on Jeju Island and southern islands, and madecassoside in Centella asiatica has been used for the treatment of skin ulcers and wounds since a long time ago.

The fermented Centella asiatica extract is a solvent extract fermented with a strain.

The Centella asiatica solvent extract is extracted through sonication under an extraction solvent. Specifically, Centella asiatica and an extractant are mixed in a weight ratio of 1: 5 to 25, preferably 1: 10 to 20, and 20 to 50 kHz, preferably Treated with an ultrasonicator with a frequency of 30 to 40 kHz and a power of 50 to 700 W, preferably 150 to 400 W, at 50 to 90° C., preferably 60 to 70° C. for 5 to 60 minutes, preferably 15 to 30 minutes it has become

When extracting Centella asiatica not by ultrasonication, but by solvent or ultra-high pressure extraction, not only a small amount of active ingredient is extracted, but also anti-obesity and anti-diabetic effects may be low.

When the weight ratio of Centella asiatica to the extraction solvent is out of the above range, the active ingredient of Centella asiatica may be extracted in a small amount in the extract.

In addition, when the frequency and power of the ultrasonicator are less than the lower limit, the active ingredient of Centella asiatica may be extracted in a small amount, and if it exceeds the upper limit, other substances in addition to the active ingredient may be extracted in a large amount, thereby reducing the effect.

In addition, when the extraction temperature and extraction time are less than the lower limit, the active ingredient of Centella asiatica can be extracted in a small amount, and when the extraction temperature and extraction time are above the upper limit, the functionality may be reduced due to heat deformation of polyphenol and alpha glucosidase. .

The extraction solvent for extracting the extract is water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof. The lower alcohol may include 20 to 99% by volume of methanol, ethanol, butanol or propanol aqueous solution, and preferably 50 to 80% by volume of ethanol aqueous solution having excellent anti-obesity effect.

Centella asiatica fermented extract of the present invention is the centella asiatica ultrasonic solvent extract, preferably, after inoculating the strain in the centella asiatica ultrasonic ethanol extract and fermented for 10 to 30 days at 25 to 50 ℃ 100 to 300 rpm.

As the strain, Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus sakei ( Lactobacillus sakei ), Leuconostoc mesenteroides ( Leuconostoc mesenteroides ) and Lactobacillus buchneri ( Lactobacillus buchneri ) at least one selected from the group consisting of and beta-When using a strain other than the strain known to produce glucosidase, the anti-obesity and anti-diabetic effect may be low or absent.

When the temperature, mixing speed, and time during fermentation are less than the lower limit, the active ingredient of Centella asiatica may be extracted in a small amount, and if it exceeds the upper limit, the desired effect may not be exhibited at all due to the decomposition of the physiologically active substances.

The fermented Centella asiatica extract of the present invention may be used in food compositions in addition to pharmaceutical compositions.

The term 'extract' used in the present invention includes an extract obtained by extracting the components contained in Centella asiatica using the solvent, a fraction fractionated therefrom, a concentrate obtained by additionally concentrating these extracts or fractions, and purified or separated purified products. , is used in the sense of including a dried product of the extract, fraction, concentrate or purified product or a powder obtained by pulverizing the same.

For the production of the purified product, various additionally performed, such as passing through an ultrafiltration membrane having a molecular weight cut-off value, or separation by various chromatography (those prepared for separation according to size, charge, hydrophobicity or affinity) A purification method may be added.

The present invention relates to a food composition for preventing or improving obesity comprising a fermented Centella asiatica extract as an active ingredient.

The 'food composition' includes, in addition to the fermented Centella asiatica extract as an active ingredient, food raw materials that can be used as food listed in the standards and specifications ('Food Code') for food commonly used in food manufacturing, and food additives described in the Food Additives Code. do.

Although not particularly limited, examples thereof include proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. The carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sugar, lactose and the like; oligosaccharides or polysaccharides such as dextrin, starch syrup, cyclodextrin and the like; Sugar alcohols such as xylitol, sorbitol, erythritol and the like can be used. As the flavoring agent, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be used.

In the case of preparing a food composition using the fermented Centella asiatica extract as an active ingredient, the fermented Centella asiatica extract does not need to be particularly limited as long as the content can prevent or treat obesity and diabetic diseases, for example, 0.1 to 99% by weight, 0.5 to 95 Weight %, 1 to 90% by weight, 2 to 80% by weight, 3 to 70% by weight, 4 to 60% by weight, 5 to 50% by weight may be included.

The fermented Centella asiatica extract, which is an active ingredient in the food composition, varies depending on the condition, weight, presence, severity and duration of the ingestion, but may be appropriately selected by a person skilled in the art. For example, it may be 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, still more preferably 20 to 800 mg, and most preferably 50 to 500 mg based on the daily dose. And, the number of administration does not need to be particularly limited, but can be adjusted by those skilled in the art within the range of 3 times a day to once a week. In the case of long-term intake for the purpose of health and hygiene or health control, it may be less than the above range.

The food composition is not particularly limited, but may be, for example, a powder, granules, tablets, capsules, pills, extracts, jelly formulations, tea bag formulations or beverage formulations.

In addition, the fermented Centella asiatica extract may be added to general food to give the function of preventing or improving obesity and diabetes, and the food that can be added is not particularly limited, but for example, the food according to Article 7 of the Food Sanitation Act. Confectionery, bread or rice cakes, cocoa processed products or chocolates, edible meat or egg products, fish meat products, tofu or jelly products, noodles, teas, coffee, beverages, special purpose foods, sauces exemplified in the standards and specifications ('Food Code') , seasoned foods, dressings, kimchi, salted fish, pickled foods, stewed foods, alcoholic beverages, raisins, and other foods. In addition, it can be added to dairy products, processed meat products, packaged meat, and processed eggs exemplified in the processing standards and ingredient specifications of livestock products according to Article 4 of the Livestock Products Sanitation Control Act ('Livestock Products Code').

On the other hand, the food composition containing the fermented Centella asiatica extract as an active ingredient may be used alone as "health functional food for preventing or improving obesity and diabetes".

The 'health functional food' refers to food manufactured (including processing) according to legal standards using raw materials or ingredients useful for the human body (Article 3, Item 1 of the Health Functional Foods Act). The term 'health functional food' may differ in terms or scope of each country, but 'Dietary Supplement' in the US, 'Food Supplement in Europe', 'Health functional food' or 'Health functional food' in Japan It may correspond to 'Food for Special Health Use (FoSHU)' and 'health food' in China.

The above food composition or health functional food may additionally contain food additives, and the suitability as a food additive is determined by the standards and standards for the item in accordance with the general rules and general test methods of the 'Food Additives Codex', unless otherwise specified. follow

In addition, in the health functional food, as a raw material announced as a 'functional raw material' or an individually recognized raw material used in "health functional food for the prevention or improvement of obesity" together with the fermented Centella asiatica extract, Lactobacillus gasseri BNR17, L-carnitine tartrate , Garcinia cambogia bark extract, conjugated linolenic acid (free fatty acid), conjugated linolenic acid (triglyceride), green mate extract, green coffee bean extract, perilla leaf extract, green tea extract, soybean germ extract, etc. purified protein), lemon balm extract mixed powder, mate hot water extract, seaweed, etc. complex extract (xantijen), fermented vinegar pomegranate complex, puer's tea extract, seomoktae (snow bean) peptide complex, vegetable oil diglyceride, wild mango seed extract , medium chain fatty acid (MCFA) containing fat, Coleus forskohlii extract, chitosan, chitooligosaccharide, green apple extract polyphenol, finger root extract powder, hibiscus complex extract, etc. It is possible to manufacture health functional food for prevention or improvement.

The present invention also provides a method for treating obesity by administering the composition to a human or non-human animal.

The present invention also provides a novel use of the fermented Centella asiatica extract for the prevention or treatment of obesity and diabetes, or for the manufacture of a medicament for animals.

The 'pharmaceutical composition' or 'medicine' may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions, in addition to the fermented Centella asiatica extract as an active ingredient.

The 'carrier' is a compound that facilitates the addition of the compound into a cell or tissue. The 'diluent' is a compound that is diluted in water to not only stabilize the biologically active form of the compound of interest, but also to dissolve the compound.

The carrier, excipient and diluent are not particularly limited, but for example, lactose, glucose, sugar, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.

The amount of the pharmaceutical composition, medicine, pharmaceutical composition for animals or veterinary medicine may vary depending on the age, sex, and weight of the patient or animal to be treated, and above all, the condition of the subject to be treated, a specific category of the disease to be treated, or It will depend on the type, route of administration, and the nature of the therapeutic agent used.

The pharmaceutical composition, medicament, pharmaceutical composition for animals or medicament for animals is appropriately selected according to the absorption rate of the active ingredient in the body, the rate of excretion, the age and weight of the patient or the animal to be treated, sex and condition, the severity of the disease to be treated, etc. , It is generally preferred to administer 0.1 to 1,000 mg/kg, preferably 1 to 500 mg/kg, more preferably 5 to 250 mg/kg, and most preferably 10 to 100 mg/kg per day. The unit dosage form formulated in this way can be administered several times at regular time intervals as needed.

The pharmaceutical composition, medicament, pharmaceutical composition for animals or medicament for animals may be administered individually as a prophylactic or therapeutic agent or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

The pharmaceutical composition, medicine, pharmaceutical composition for animals or medicine for animals may be formulated into oral dosage forms such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc. have. In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, and such solid preparations include at least one excipient to the compound, for example, starch, calcium carbonate, sugar or lactose, gelatin. It can be prepared by mixing and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

The method for treating obesity and diabetes is to administer the composition to a human or non-human animal, particularly a mammal, for example, administering the composition to an obese subject to be treated.

The dosage, administration method, and frequency of administration for the treatment may refer to the dosage, administration method, and frequency of administration of the pharmaceutical composition, medicine, pharmaceutical composition for animals or medicament for animals.

Hereinafter, preferred examples are presented to help the understanding of the present invention, but the following examples are merely illustrative of the present invention, and it will be apparent to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It goes without saying that such variations and modifications fall within the scope of the appended claims.

Example 1. Centella asiatica fermented extract

culture medium

Leukonostok mecenteroides was inoculated with 1% MRS broth and cultured at 37° C. for 24 hours to be used as a culture medium.

fermented extract

Centella asiatica and 50% by volume ethanol aqueous solution were mixed in a weight ratio of 1:10, put into an ultrasonicator (JAC Ultrasinic, Hwaseng, Korea), and then extracted for 30 minutes using ultrasonic waves (40 kHz, 250 W) at 60 ° C. Centrifugation at rpm for 5 minutes to recover the supernatant to obtain an ultrasonic ethanol extract of Centella asiatica. After the obtained Centella asiatica ultrasonic ethanol extract was cooled at room temperature, the Centella asiatica ultrasonic ethanol extract and Leukonostok mecenteroides were mixed in a weight ratio of 1: 2 to adjust the pH to 6.95 to 7.05, and then 1 weight per 100 parts by weight of the mixture After fermenting at 37 ℃ for 48 hours, crushed using a homogenizer (HG-15A, DAIHAN, Korea), adjusted to pH 4.78±0.05, fermented at 45 ℃ for 48 hours, and then centrifuged to recover the supernatant. An extract was obtained.

Example 2. Centella asiatica fermented extract_ultrasound omitted

culture medium

Leukonostok mecenteroides was inoculated with 1% MRS broth and cultured at 37° C. for 24 hours to be used as a culture medium.

fermented extract

Centella asiatica and 50% by volume ethanol aqueous solution were mixed in a weight ratio of 1:10 and extracted for 50 minutes at 80 ° C. Then, the extract was centrifuged at 1000 rpm for 5 minutes to recover the supernatant, thereby obtaining an ethanol extract of Centella asiatica. After cooling the ethanol extract of Centella asiatica obtained above at room temperature, the ethanol extract of Centella asiatica and Leukonostok mecenteroides were mixed in a weight ratio of 1: 2 to adjust the pH to 6.95 to 7.05, and then 1 part by weight of sugar was added to 100 parts by weight of the mixture. and fermented at 37 ° C for 48 hours, then crushed using a Homogenizer (HG-15A, DAIHAN, Korea), adjusted to pH 4.78 ㅁ 0.05, fermented at 45 ° C for 48 hours, and then centrifuged to recover the supernatant to obtain a fermented Centella asiatica extract. obtained.

Comparative Example 1. Ultrasonic Extract

Centella asiatica and 50% by volume ethanol aqueous solution were mixed in a weight ratio of 1:10, put in an ultrasonicator (JAC Ultrasinic, Hwaseng, Korea), and then extracted for 30 minutes using ultrasonic waves (50 kHz, 700 W) at 80 ° C. Centrifugation at rpm for 5 minutes to recover the supernatant to obtain an ultrasonic centrifugal extract.

<Test Example>

Test Example 1. Measurement of polyphenol content and antioxidant activity

1-1. Polyphenol content (mg GAE/g DW): Polyphenol is a secondary metabolite of plants, and its representative physiologically active functions include anti-mutation, anti-cancer, anti-inflammatory and anti-obesity. It is known that it has anti-obesity functions by delaying the digestion and decomposition of carbohydrates and lipids by inhibition.

The Folin-Denis method was modified and measured, and the molybdenum blue color reaction generated by adding a folin-ciocalteu's phenol solution (Folin &Ciocalteu'sphenol; Sigma-Aldrich, USA) to the sample was reduced by a polyphenol compound was used as a principle. After adding 0.2 N FC solution to 0.14 ml of the sample and leaving it for 10 minutes, 0.56 ml of 7.5% Na ₂ CO ₃ was added and reacted for 1 hour, and the absorbance value was measured at 756 nm. Gallic acid (Sigma-Aldrich, USA) was used as a standard material, and the unit was expressed as mg gallic acid equivalent (GAE)/g dry weight (DW) compared with the prepared gallic acid calibration curve.

1-2. Antioxidant activity (%): After adding 0.1 M DPPH (1,1-Diphenyl-2-picrylhydrazyl) solution to the extract, the color was developed in a dark room for 20 min. Absorbance was measured at a wavelength of 517 nm, and ascorbic acid was used as a control. Therefore, the antioxidant capacity was expressed as a percentage using the formula below based on the absorbance of the control group.

[Equation 1]

Radical scavenging activity (%)=[(1-absorbance of sample)/absorbance of control group)]X100

division Example 1 Example 2 Comparative Example 1 polyphenols
(mg GAE/g DW) 28.10±1.12 21.11±0.85 17.15±0.63 antioxidant capacity
(%) 42.26±2.46 34.15±1.20 25.53±1.43

As shown in Table 1 above, it was confirmed that the fermented Centella asiatica ferment extract prepared according to Examples 1 and 2 of the present invention was superior to Comparative Example 1 in the content of polyphenols known to have anti-obesity function and antioxidant activity.

Test Example 2. α-glucosidase inhibitory activity (GAI) and lipase inhibitory activity (LAI) measurement

2-1. α-glucosidase inhibitory activity (%): α-glucosidase inhibitors are distributed in some plants and microorganisms and are substances that inhibit the decomposition of oligosaccharides into monosaccharides. Inhibition of α-glucosidase activity inhibits the smooth absorption of saccharides in the intestine, thereby preventing obesity and suppressing postprandial blood glucose rise.

α-glucosidase and potassium phosphate buffer (pH 6.8) were mixed in 50 μL of the sample and reacted at 37 °C for 15 minutes, then the substrate p-nitrophenyl α-D-glucopyranoside was added and reacted at 37 °C for 10 minutes. The reaction was stopped by adding 0.1 M NaOH to the reaction solution, and the absorbance was measured at 405 nm to measure p-nitrophenol. For activity comparison, acarbose was used as a positive control.

2-2. Lipase inhibitory activity (LAI) (%): Lipase secreted from the body is an enzyme that hydrolyzes triacylglycerol, a fat, into 2-monoacylglycerol and fatty acid when ingested. It is absorbed from the mucosal cells of the small intestine and used as an energy source and a component in the human body. It is possible to prevent obesity through the accumulation of fat in the body through excretion by preventing the decomposition of triglyceride and absorption by inhibiting the activity of lipase, which is a major enzyme in fat metabolism.

For LIA measurement, 10 mM MOPS, 1 mM EDTA (pH 6.8), and tris buffer were added to porcine pancreatic lipase and reacted at 37 °C for 15 minutes. After the reaction, p-nitrophenyl butyrate was added and the reaction was carried out again at 37 °C, and the degree of hydrolysis of P-Nitrophenyl butyrate to p-nitrophenol was measured by absorbance. At this time, orlistat was used as a positive control for activity comparison.

division Example 1 Example 2 Comparative Example 1 α-glucosidase inhibitory activity (%) 57.48±3.22 49.66±1.29 37.02±1.30 Lipase inhibitory activity (%) 61.59±2.06 57.80±2.02 44.42±2.82

As shown in Table 2 above, it was confirmed that the fermented Centella asiatica ferment extract prepared according to Examples 1 and 2 of the present invention had superior α-glucosidase inhibitory activity compared to Comparative Example 1. If the activity of lipase of alpha-amylase, which acts as a key enzyme in carbohydrate metabolism, is inhibited, polysaccharides cannot be decomposed and fat accumulation in the body can be prevented, thereby preventing obesity. Therefore, it is judged that the fermented Centella asiatica extract of Examples 1 and 2 has a high possibility of preventing obesity by inhibiting and controlling monosaccharide production through effective α-glucosidase inhibition.

In addition, since obesity is caused by the accumulation of triglyceride (TG) in adipocytes by the differentiation and adipogenesis process of preadipocytes, it is known that controlling the adipogenesis mechanism is an effective method for the treatment of obesity. Since the fermented Centella asiatica ferment extract prepared according to Examples 1 and 2 has superior lipase inhibitory ability compared to Comparative Example 1, it is effective in treating obesity.

Test Example 3. Anti-obesity protein expression confirmation (Western-blotting)

After culturing and differentiation of 3T3-L1 cells, lysis buffer was added to lyse them, and then the protein was quantified using the Bradford method, the protein concentration was adjusted to the same level, and electrophoresis was performed using SDS-PAGE. Proteins separated by electrophoresis are transferred using transfer buffer using a transfer membrane, blocked with 5% non-fat skim milk solution, and then reacted with primary antibodies GLUT4, PPAR-α, β-actin to the membrane. It was treated with an ECL detection kit and developed by exposure to an X-ray film to compare protein expression levels.

3T3-L1, a pre-adipocyte, accumulates triglycerides in the cell due to various transcription factors and hormones that form adipocytes. PPAR-α protein is expressed in muscle and hepatocytes and functions to regulate beta oxidation in mitochondria. Recently, it has been reported that weight loss is induced by suppressing appetite by forming a feeling of satiety, and PPAR-α is recognized as a major protein in anti-obesity action in the development of an anti-obesity agent. In addition, the expression of glucose transporter 4 (GLUT4), which moves glucose in the blood into cells, promotes diffusion of circulating glucose from the cell surface to muscle and fat cells in a concentration gradient, thereby promoting the consumption of sugar and controlling blood sugar. It plays an important role in controlling obesity and blood sugar.

1 is a view showing the expression of glucose transporter 4 (GLUT4) in a fermented Centella asiatica extract prepared according to Example 1 of the present invention.

As shown in Figure 1, the fermented extract of Centella asiatica of Example 1 affects the expression of GLUT4 (glucose transporter 4), which plays a role in delivering glucose in the blood into cells, and plays an important role in the insulin signaling system. When transporter 4 (GLUT4) is disrupted, hyperglycemia is caused by the accumulation of blood glucose in the blood.

When the effect of the fermented Centella asiatica extract of Example 1 on GLUT4, which is a major protein, when moving glucose in the blood into cells was confirmed, it was confirmed that the expression of GLUT4 increased as the concentration of the fermented Centella asiatica extract of Example 1 increased. Therefore, the fermented Centella asiatica extract of Example 1 has anti-obesity and hypoglycemic effects by decreasing the activity of enzymes related to digestion of carbohydrates and lipids as well as increasing the expression of GLUT4 protein.

2 is a view showing the expression of PPAR-α of the fermented Centella asiatica extract prepared according to Example 1 of the present invention.

PPAR-α protein is expressed in muscle and hepatocytes and functions to regulate beta oxidation in mitochondria. Recently, there has been a report that it induces weight loss by suppressing appetite by forming a feeling of satiety, and PPAR-α is recognized as a major protein in anti-obesity action in the development of an anti-obesity agent.

As shown in Figure 2, for the evaluation of the anti-obesity mechanism of the fermented Centella asiatica extract of Example 1, the fermented Centella asiatica extract of Example 1 was treated at 0.0-0.8 mg/mL and the level of PPAR-α expression after cell culture was assessed by Western blotting. was confirmed using . As a result, PPAR-α expression is increased depending on the concentration of the fermented extract of Centella asiatica of Example 1, so there is an anti-obesity effect.

The fermented Centella asiatica extract of Example 1 activates fat beta oxidation by increasing the expression of PPAR-α to promote fatty acid metabolism, and inhibits fat differentiation and reduces lipid content by reducing blood sugar through GLUT4 expression. It was confirmed to have anti-obesity and anti-diabetic effects.

Hereinafter, a formulation example of a composition comprising the extract of the present invention will be described, but the present invention is not intended to limit the present invention, but merely to describe it in detail.

Formulation Example 1: Preparation of powder

20 mg of extract powder of Example 1

Lactose 100 mg

talc 10 mg

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

Formulation Example 2: Preparation of tablets

10 mg of extract powder of Example 1

100 mg cornstarch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

Formulation Example 3: Preparation of capsules

10 mg of extract powder of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg magnesium stearate

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

Formulation Example 4: Preparation of granules

1,000 mg of extract powder of Example 1

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

50 μg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

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

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is mixed with ingredients suitable for health functional food in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the ingredients are mixed according to a conventional health functional food manufacturing method. Next, the granules can be prepared and used in the preparation of a health functional food composition according to a conventional method.

Formulation Example 5: Preparation of beverage formulations

1,000 mg of extract powder of Example 1

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to a conventional beverage manufacturing method, after stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and then refrigerated. used in the manufacture of functional beverage compositions.

Claims (7)

Centella asiatica A pharmaceutical composition for preventing or treating obesity and diabetes, characterized in that it contains a fermented extract as an active ingredient. The pharmaceutical composition for preventing or treating obesity and diabetes according to claim 1, wherein the fermented Centella asiatica extract is a fermented extract obtained by fermenting the Centella asiatica solvent extract as a strain. According to claim 2, wherein the strain is Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus sakei ( Lactobacillus sakei ), Leuconostoc mesenteroides ( Leuconostoc mesenteroides ) and Lactobacillus buchneri ( Lactobacillus buchneri ) group consisting of A pharmaceutical composition for the prevention or treatment of obesity and diabetes, characterized in that at least one selected from. The pharmaceutical composition for preventing or treating obesity and diabetes according to claim 2, wherein the centella asiatica solvent extract is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. [Claim 3] The pharmaceutical composition for preventing or treating obesity and diabetes according to claim 2, wherein the centella asiatica solvent extract is treated for 5 to 60 minutes with an ultrasonicator of a frequency of 20 to 50 kHz and a power of 50 to 700 W. [Claim 6] The pharmaceutical composition for the prevention or treatment of obesity and diabetes according to claim 5, wherein the extraction temperature during the ultrasonic treatment is 50 to 90 °C. Centella asiatica Food composition for preventing or improving obesity and diabetes, characterized in that it contains a fermented extract as an active ingredient.

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