KR100733336B1 - Composition for Prevention or Treatment of Obesity or Hyperlipidemia Comprising Fermentation Product of Puerariae Radix or Puerariae Flos Extract - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of obesity or hyperlipidemia containing lactic acid bacterium fermented product of the root or brownish extract as an active ingredient, more specifically, hot water extraction after adding water to the root or browned and lactic acid bacteria to the water extract It relates to a pharmaceutical composition and health food for the prevention or treatment of obesity or hyperlipidemia containing lactic acid bacteria fermented product of the root or browned water extract prepared by inoculating fermentation. Fermented extract of brown root or brown leaf extract has an excellent effect on obesity and hyperlipidemia that cannot be obtained by brown root or brown leaf extract alone, and it is highly purified to apply to human body without using organic solvents such as alcohol, especially toxic methanol. Since it is not necessary, it can be usefully used for the prevention or treatment of obesity and hyperlipidemia.

Brown root, browning, extract, fermented product, obesity, hyperlipidemia

Description

Composition for prevention or treatment of obesity or hyperlipidemia Comprising Fermentation Product of Puerariae Radix or Puerariae Flos Extract}

Figure 1 is a graph showing the effect on the growth of MCF-7 cells after the fermented product of the root (A) and brown (B) water extract fermented by different lactic acid bacteria at a concentration of 6 ㎍ / ㎖.

Figure 2 is a graph showing the effect on the growth of MCF-7 cells after the addition of fermented products of the root (A) and brown (B) water extract fermented by different lactic acid bacteria at a concentration of 25 ㎍ / ㎖.

Figure 3 is a graph showing the change in weight over time of the test animal extracted ovary.

Figure 4 is a photograph showing whether the atrophy of the uterus occurred after administration of the sample to the test animal.

Figure 5 is a photograph showing the change in body fat after administration of the sample to the test animal.

The present invention relates to a composition that can be effectively used for obesity or hyperlipidemia, and more particularly to a composition for the prevention or treatment of obesity or hyperlipidemia containing the microorganism fermentation of the extract of the root or browning as an active ingredient.

Obesity refers to a condition in which energy intake and consumption are not balanced and excess energy is accumulated as fat, resulting in abnormally high body fat and metabolic abnormalities. Obesity is a serious health problem in Korea as well as in the West, where meat is a staple food, and about 30-40% of modern people are known to have obesity. The causes of obesity include the lack of exercise and other endocrine and pathophysiological abnormalities due to the development of civilization in pursuit of high fat and high calorie diet patterns and convenience according to changes in the environment and society. In addition, the fact that genetic factors act as an important cause of obesity can be seen in the fact that obesity is associated with households, and in addition to environmental factors, genetic factors affect at least 30-50% of the effects of obesity. It is known to be crazy.

Obesity affects all age groups, especially in older adults. In the Orient, there was not much obesity due to vegetarian-oriented lifestyle, but in modern times, obesity is gradually increasing in Asians as the dietary habits and the living environment change due to the influence of Western culture. In addition, modern man has been reported that physical strength is reduced due to overwork, heavy drinking, stress, etc., about 30-40% have obesity, about 10% of them have been reported severe or pathological obesity.

Obesity is classified into simple obesity and secondary obesity (symptomatic obesity). The causes of simplicity obesity include overeating, lack of exercise, and decreased metabolism. Most cases of clinically judged obesity are simplicity obesity, and when the obesity manifests and persists, it causes various health disorders. Secondary obesity is obesity caused by any underlying disease, and may include endocrine obesity, hypothalamic obesity, hereditary obesity, or obesity due to drugs. Obesity is a complication due to various diseases, but obesity itself causes diabetes, hypertension, cardiovascular disease, pseudotumor cerebri, hyperlipidemia, sleep apnea, cancer, pulmonary hypertension, cholecystitis, and osteoarthritis. It is known to cause various diseases or cause problems such as deactivation of activity.

Obesity patients usually lose weight through diet and exercise, and other behavioral, psychotherapy, drug or surgical therapies (such as atrophy) are combined. Diet is a treatment method that controls the total energy intake, but the expected metabolic rate decreases, so weight loss cannot be achieved, and even if the weight loss is successful, Or physical and mental pains such as malnutrition, fasting or stress. Exercise therapy not only increases energy consumption, but also improves metabolic rate at rest, corrects insulin resistance, and decreases body fat.However, aerobic exercise for 20 minutes or more is performed three times a week. It is not easy to carry out continuously because it is necessary. Behavioral or psychotherapy is a way to maintain diet or exercise therapy, but it is generally difficult to achieve sufficient effects.

When the above treatment method is not effective or when urgent treatment for extreme obesity is necessary, drug therapy or surgical therapy is performed, but the surgical therapy is very burdensome to the patient. The anti-obesity drugs used for drug therapy include appetite suppressants, digestive absorption inhibitors, obesity-inhibiting agents, or metabolic accelerators, but these anti-obesity drugs have a risk of causing side effects such as drug dependence. In addition, there is a problem in that resistance can be developed in a short period of time to a patient to be administered, so that it cannot be used continuously for a long time. In addition, in the case of exercise therapy, if the obese person is forced to violent exercise, the risk of injury to the lower extremity joint increases, so it is generally preferable to perform light swimming. In this case, activating energy metabolism, increasing muscles by performing light exercise, such as a drug with a low side effect and light swimming, and promoting energy consumption by increasing basic metabolism are considered as one preferable method for improving obesity.

Therefore, there is an urgent need for the development of new methods or medicines that can be used continuously for a long period of time without burdening the human body, and which effectively promote weight loss.

On the other hand, the root roots are dried roots by removing the bark of the perennial herb, Pueraria lobata Ohwi, which is a herb that grows in the mountains of Korea, China, and Japan. In Chinese medicine, the various parts of the 칡 are classified into dry roots, roots, brown flowers, brown or brown fruit, brown leaves, brown leaves, and vines, which are used differently depending on the symptoms and treatment methods of the disease.

In the oriental medicine, the roots are used to relieve the muscles, so it is used for the purpose of releasing cold body swelling, rash, anti-alcoholic pain, etc. (Herbology, 148, 1995). In ideology, it is classified as a drug of Taeumin and is used for basic prescriptions such as Galeunhaegitang, thermal Hantang, Galgunseunggitang, Carved Daehwangtang, and Galgunbupyeongtang (Theory of Thought, 343-50, 1985).

Brown root contains a large amount of isoflavonoides (Chinese modern research in clinical applications, 626, 1994), and studies on the components mainly related to isoflavonoids have been reported. Pharmacological actions associated with carpal roots include antidipsotropic action (Proceeding of National Academy of Sciences of USA, 95 (5): 2198-203, 1998; Alcoholism: Clinical & Experimental Research 40 (4): 659-63, 1996), hepatoprotective action (Planta Medica, 64 (5): 413-6, 1998), hangover inhibitory action (Proceeding of National Academy of Sciences of USA, 94 (5): 1675-9, 1997), antioxidant activity ( Chem. Pharm. Bull. 40 (3): 721-4, 1992). In addition, the relaxation of smooth muscle, coronary artery dilation and blood pressure drop in the cardiovascular system, cardiac rhythm control, platelet aggregation inhibitory effect, hyperglycemic action, antipyretic action are known (Chinese modern research clinical application, 626-632, 1994). It is also applied to headaches, antiganglia, limb sensory disturbances, lons, and diabetes in old age diseases (Anti-Blood Pharmacology, 186-187, 1989). However, until now, there have been no reports of using rooting or browning directly for alleviating symptoms such as obesity and hyperlipidemia.

It is an object of the present invention to provide a composition for the prevention or treatment of obesity or osteopathy, which contains lactic acid bacteria fermented product of an extract of brown root or gallium as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of obesity or hyperlipidemia, containing the lactic acid bacteria fermented product of the root or brown extract as an active ingredient.

In another aspect, the present invention provides a health food for the prevention or treatment of obesity or osteopathy containing lactic acid bacteria fermentation of the extract of the roots or browning as an active ingredient

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for the prevention or treatment of obesity or hyperlipidemia containing lactic acid bacteria fermented product of the water extract prepared by extracting the root or browning with water as an active ingredient.

In the above, in order to prepare lactic acid bacteria fermented product of the root or browned water extract (hereinafter, abbreviated as "rooted fermented product" and "browned fermented product" respectively), first, the rooted or browned, preferably dried rooted or browned 2 After adding 10 times the amount of water to prepare a water extract extracted from hot water at 70 to 100 ℃. After cooling the extract, the lactic acid bacteria were inoculated at 1 to 5% by weight, preferably 2% by weight of the sample weight, and fermented at 30 to 40 ° C, preferably at 37 ° C for 24 to 52 hours, preferably 48 hours. Brown or brown fermentation products are prepared.

In the above, as the lactic acid bacteria, various strains or commercially available lactic acid bacteria can be used without limitation. Specifically, Bifidobacterium, Lactobacillus, Streptococcus, Leukonostock, Pediococcus and Lactococcus microorganisms, preferably Bifidobacterium breve, Bifidobacterium longugum (B. longum), Bifidobacterium infantis, Latobacillus bulgaricus, L. acidophilus, Streptocuccus thermophilus, Streptocuccus S. faecium, S. faecalis, Leuconostoc mesenteroides, Pediococcus cerevisiae and Clostridium butyricum It may be selected from among, it is preferable to use the microorganism of the genus Bifidobacterium.

The rooted or browned fermented product may be administered orally or parenterally during clinical administration and may be used in the form of a general pharmaceutical formulation. That is, the rooting or grinding fermentation product of the present invention can be administered in various oral and parenteral formulations during actual clinical administration, and when formulated, fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used, are formulated. It is prepared using diluent or excipient. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, It is prepared by mixing gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

Dosage units may, for example, contain 1, 2, 3 or 4 times the individual dose, or 1/2, 1/3 or 1/4 times. Individual dosages contain amounts in which the effective drug is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dose.

The human dose of the rooting or browning fermentation product is appropriately selected according to the absorption rate, inactivation rate and excretion rate of the active ingredient in the body, the age, sex, condition, degree of disease, etc. of the patient, 10 to 300 mg / kg to adults It is preferably 20 to 100 mg / kg, it may be administered 1 to 6 times a day divided.

As a result of conducting oral toxicity and the intraperitoneal administration of the rooted or browned fermented product to mice, all 50% of the lethal dose (LD ₅₀ ) by oral toxicity test was at least 5 g / kg or more as a safe substance. It turned out.

In a preferred embodiment of the present invention, in order to select the lactic acid bacteria strains that are most effective in producing the root or brown fermentation products, preliminarily selected strains having excellent activity of β-glucosidase among 30 kinds of lactic acid bacteria and then the roots and browning Fermentation was carried out to determine the estrogen phase activity. As a result, it was confirmed that when the fermented roots and browning fermented products were significantly increased in vitro, the estrogen phase activity was higher, and the estrogen-like activity was higher in the browned fermented products than the roots (see Fig. 1). In addition, as a result of experiments with various lactic acid bacteria, it was confirmed that the largest estrogen-phase activity in Bifidobacterium K111 among them (see Fig. 2).

Next, the effect of the rooting or browning fermentation on obesity or hyperlipidemia was examined using test animals from which ovaries were removed. Continuous oral administration of the test substance did not show any particular clinical signs and deaths, but the weight of the ovary-removed test animals was significantly increased from 1 week compared to the control group. It is thought that the body weight of the test animal is increased due to the rapid change of hormone caused by ovarian extraction. However, the weight gain was significantly reduced when the root fermentation was administered (see FIG. 3). It is believed that this is because the rooting fermentation product has a similar action to estrogen, thereby inhibiting weight gain.

As autopsy findings, uterine atrophy due to ovarian extraction was clearly observed in the negative control group, but the atrophy of the atrophic fermentation group was significantly reduced compared to the negative control group (see FIG. 4). In addition, spleen organs were increased in the ovarian extraction group compared to the normal control group, but the enlarged organs were significantly decreased when the oral root fermentation was administered orally (see Table 3). This means that the rooting fermentation has an estrogen replacement effect against changes in the uterus and spleen organs due to estrogen deficiency.

Abdominal fat was significantly increased in the ovary-negative negative control group than in the normal control group, but significantly decreased in the case of the administration of the root fermentation product. In the negative control group, the amount of abdominal fat was increased by more than 250% compared to the normal control group, but when the fermented roots were administered, the abdominal fat was reduced by 20% compared to the normal control group (see FIG. 5 and Table 4). In addition, the estrogen content of the ovary extraction was reduced by more than 20% compared to the normal control group, but increased slightly compared to the normal control group in the carp fermentation group (see Table 5).

Stress, aging, pregnancy and hormonal imbalances have a large impact on cholesterol levels. When administration of brown or brown fermented products, cholesterol concentration was significantly reduced compared to the negative control group. From this, it was speculated that the browned fermented products promote the metabolism of cholesterol, which is a precursor of steroid hormone and sex hormone, into estrogen. In addition, the amount of LDL-cholesterol and triglycerides, which are important causes of atherosclerosis, were reduced by more than 30% compared to the negative control group (see Table 6). In other words, all kinds of cholesterol were statistically significantly reduced when the root fermentation product was administered.

According to the present invention, it was shown that obesity and hyperlipidemia were prevented and treated effectively when the fermented roots were administered to the phytoestrogens of phytoestrogens, which were higher than the browned fermented products. In addition to increased phytoestrogens, it can be predicted that certain potent ingredients that give synergistic effects in the prevention and treatment of obesity and hyperlipidemia have increased. Therefore, the composition of the present invention is considered to act as a substance for maintaining a certain amount of hormone balance, and is expected to be useful for preventing and treating various endocrine diseases as well as obesity and hyperlipidemia.

In addition, the present invention provides a health food for the prevention or treatment of obesity or hyperlipidemia, containing the root or brown fermented product as an active ingredient.

The rooted or browned fermented product may be added to a health food for the purpose of improving obesity or hyperlipidemia, and when the rooted or browned fermented product is used as a food additive, these may be added as it is or used with other food or food ingredients, and It can be suitably used according to the phosphorus method. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, in the preparation of foods or beverages, brown or brown fermentation products are added in an amount of 30% by weight or less, preferably 10% by weight or less based on the raw materials. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .

There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. Natural carbohydrates described above include glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of rooted or browned fermentation.

In addition, the rooting or browning fermentation products are various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol And carbonation agents used in carbonated beverages. In addition, the root root fermentation may contain a flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is usually selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the rooting or grinding fermentation product.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Example 1 Preparation of Root Fermentation

<1-1> Root extract of hot water

Domestic dried roots were purchased and used as a sample. 5 L of water was added to 1 kg of the selected roots, followed by hot water extraction at 100 ° C. to obtain an extract.

<1-2> Strain Culture

Bifidobacterium breb K-111 (Kim, Kyung Hee University), Bifidobacterium brb K-110 (KCCM 10097) and Streptococcus Passium (KCTC 3077) strains were selected first. Selected isolates and commercially available Lactobacillus, Bifidobacterium, and Streptococcus microorganisms were passaged in standard medium (aerobic bacteria; MRS medium, anaerobic bacteria; GAM medium), and the initial bacterial count was adjusted to 10 ⁸ Used for.

<1-3> Preparation of Water Extract Fermentation (1)

After cooling the water extract of the root roots prepared in Example <1-1>, 50 g (wet weight) of the lactic acid bacteria incubated in the standard medium of Example <1-2> was added and reacted at 37 ° C. for 6 to 96 hours. And lyophilized to obtain the rooted fermented powder of the present invention.

<1-4> Preparation of Water Extract Fermentation (2)

After cooling the water extract of the roots prepared in Example <1-1>, the lactic acid bacteria cultured in the standard medium of Example <1-2> were inoculated with 1% of the weight of the sample of the roots root extract, and at 37 ° C, 36. After fermentation for a time lyophilized to prepare the powder of the brown root fermentation of the present invention.

Example 2: Preparation of Fermented Water of Extractive Water

Except for using browning instead of brown root, the same method as in Example 1 was prepared to prepare a powder of fermented lyophilized water extract.

Comparative Example 1: Preparation of Fermented Soybean Extract

Except for using soybeans instead of the roots was treated in the same manner as in Example 1 to prepare a fermented product of the lyophilized soybean water extract (hereinafter referred to as "soybean fermented product") powder.

Comparative Example 2: Preparation of Root, Brownled and Soybean Extract

In the same manner as in Example <1-1>, water extracts of brown, brown and soybean were prepared, and then lyophilized to prepare their water extract powders.

Experimental Example 1: Analysis of Isoflavonoid Components of Root Extract and Fermentation thereof

The fermented root extract of the root extract obtained in Example <1-1> and the root extract of the extract produced in Example <1-3> (Hereinafter, unless otherwise stated, the fermented root of Example <1-3>) The content of isoflavones contained in and acting as phytoestrogens was analyzed using HPLC (Mitsubishi Corporation, Japan). Specifically, a column of 4.6 × 250 mm size was used, and a solvent in which acetonitrile containing 0.1% acetic acid and secondary distilled water containing 0.1% acetic acid was mixed in a 30:70 ratio was used as the mobile phase. The injection amount of the sample was 10 µl, and the flow rate was adjusted to 1.0 ml / min. The content of isoflavonoids contained in the sample was detected by a UV detector at a wavelength of 254 nm, and the isoflavone standard was used by Sigma, USA. The results are shown in Table 1.

Changes in Isoflavonoid Contents during Lactic Acid Bacteria Fermentation (Unit: ㎎ / dry powder 100 g) Fermentation time component 0 12 24 36 48 Puerin 1606.0 1619.0 1615.0 1624.2 1665.4 DIDJIN 774.6 - - - - Glycidine 237.1 - - - - Jannistin 24.9 - - - - Marronyl Dyedjin 197.0 - - - - Here - - - - 207.1 Dyed Jane 476.7 3545.9 3759.2 3849.4 3850.4 Calcide - 259.3 278.9 270.2 273.4 Genistein - 36.8 46.9 66.4 48.2

From the above results, it is confirmed that the fermented product of the present invention causes a lot of changes in the isoflavonoid component due to lactic acid bacteria fermentation, and in particular, the contents of the active ingredients Dyzezein and Genistein are greatly increased.

Experimental Example 2: Determination of Estrogen Effects on Water Extracts and Fermentations

Estrogen effect was measured using the fermented product of the present invention MCF-7 cell line breast cancer cell line.

To this end, MCF-7 cells were _first cultured under conditions of 5% CO ₂ , 37 ° C. using DMEM medium (Dullbecco's modified Eagle's medium, Sigma, USA) containing 10% FBS. Cells were washed with PBS buffer prior to hormone induction and cells for 2 days in phenol red-free DMEM containing 10% CD-FBS (charcoal-dextran stripped FBS) to remove all estrogen sources prior to sample processing. Was incubated. The cultured cells were treated with 0.25% trypsin-EDTA and centrifuged at 1,000 rpm for 5 minutes, and 0.1 ml of the cells were measured in 96-well culture plates to measure 5 × 10 ³ cells / well. After 24 hours, the samples were treated with 6 μg / ml and 25 μg / ml, respectively, and cultured for 6 days (144 hours), and then the proliferation of the cells was measured by MTT assay. At this time, the sample was used by dissolving the ethyl acetate fraction of the fermented water and the extract of brown and brown water extract prepared in Examples and Comparative Examples in DMSO so that the final concentration is 0.1% or less.

For MTT assay, 50 μl of 2 mg / ml of MTT reagent was added to the cells incubated for 6 days and reacted in an incubator for 30 minutes. After removing the medium, 100 μl of DMSO (dimethyl sulfoxide) was added thereto, and the absorbance was measured using an ELISA reader at a wavelength of 540 nm.

As a result, when the sample was used at a concentration of 6 ㎍ / ㎖, the estrogen effect was significantly increased when the extract was fermented than the simple water extract (G, K) in both rooting and browning, and the overall estrogenous effect was browning than the rooting. It was confirmed that the excellent (Fig. 1). In addition, as Lactic acid bacteria, Bifidobacterium K111 (G111, K111) was found to increase the estrogen-phase efficacy most.

On the other hand, even when a high concentration of samples was used, the overall estrogen effect was superior to browning than browning, and in case of browning, Bifidobacterium K110 was used as lactic acid bacterium, and in the case of browning, Bifidobacterium K111 was used. In this case, the estrogen phase effect was greatly increased (FIG. 2).

Experimental Example 3: efficacy test for obesity and hyperlipidemia using experimental animals

<3-1> Establishment of Osteoporosis Induced Model and Visual Observation

<3-1-1> Establishment of Osteoporosis Induction Model

The present inventors investigated the effect of the rooting or browning fermentation product prepared in the above example on obesity and hyperlipidemia in an animal model in which ovary was removed and osteoporosis was induced. Osteoporosis-induced animal model is postmenopausal Type I. Experimental animals were used to purchase 8-week-old SD (Sprague-Daweley) rat females from Orient Co., Ltd. Osteoporosis was induced by ovary at 9 weeks of age, followed by an experiment at 10 weeks of age after acclimatization for 1 week. At this time, the group without ovaries was used as a normal control group and the group without ovaries and no drug treatment was used as a negative control group.

Specifically, anesthetics (ketamine hydrochloride 50 mg / ml, Yuhan + xylazine 20 mg / ml, Bayer Korea Co., Ltd.) were subcutaneously injected into the back of the test animal for general anesthesia, and the hair of the lower abdomen was removed. The ovaries were removed after disinfection of the surgical site with iodine (Samil Pharmaceuticals) while the animals were lying on their backs. The skin, abs and peritoneum were incised about 1 cm from the lower abdomen around the midline of the test animal and the ovaries were exposed by sterile tweezers to ligation of the fallopian tubes, and the left and right ovaries were removed. To prevent infection, 0.4 ml of antibiotic (Jampaporte-4, Unichem Co., Ltd.) was injected intraperitoneally and the peritoneum, abs and skin were sutured with silk thread.

<3-1-2> Administration of Sample

A sample of 5 g / kg of dry content (sample amount before extraction) prepared in Examples and Comparative Examples was dissolved in 10 ml / kg of physiological saline and used in an animal experiment as a dosage. The number of test animals per test group and the dosage of test substance are shown in Table 2 below. Sterilized physiological saline (10 ml / kg) was administered to the normal and negative controls. Experimental animals were fixed by the dorsal skin fixation method, and forced oral administration into the stomach using metallic sonde and injection tube for oral administration. Dosing period was administered once a week for 7 weeks and 14 weeks, the dosage was calculated based on the body weight measured once a week for the dosage for each group.

group Animal count (horses) Animal Number Dosage amount (g / kg) Dose (ml / kg) C 15 1 to 15 0 10 N.C 15 16-30 0 10 T1 15 31-45 5 10 T2 15 46-60 5 10 T3 15 61-75 5 10 T4 15 76-90 5 10

C; Normal control, N.C; Negative control, T1; Soy extract administration group, T2; Soybean fermented group, T3; Reed fermentation group, T4; Group of fermentation fermentation.

<3-1-3> Weight change measurement

All animals used in the test were weighed at 10 am once a week to investigate the effect of the administration of the sample on the weight change of the test animals. All experimental groups were weighed using an animal weighing scale (Mettler, AT261) and the mean and standard deviation were calculated. Statistical significance between the control and test groups was tested by Student's t-test in all the experiments below. At this time, p <0.05 was determined as a significant difference between the experimental groups.

As a result, the weight of the negative control group from which the ovary was extracted increased significantly (p <0.01) from 1 week compared to the normal control group, but the weight gained from the continuous administration of the rooting fermented products for 2 weeks decreased significantly (p <0.01). It was confirmed that (Fig. 3).

<3-1-4> Autopsy Findings

At 7 and 14 weeks after sample administration, the lower abdomen was opened under ether anesthesia, and blood was collected from the abdominal aorta, and internal organs were visually observed. After the blood collection, the blood was bleeding and the weights of the uterus, brain, pituitary gland, spleen, adrenal gland, kidney, liver, lung and heart were measured using a scale.

As a result, no abnormal findings related to the administration of test substance were observed in all experimental groups except that uterine atrophy due to ovarian extraction was observed. In the negative control group, the uterus was remarkably contracted as compared with the uterus of the normal control group. However, when the root fermentation product of the present invention was administered, uterine atrophy was significantly suppressed compared to other sample administration groups (FIG. 4). It was confirmed that the atrophy of the uterus was significantly suppressed compared to other experimental groups, although it was not normal even from the organ weight (Table 3).

gender female Experimental group C N.C T1 T2 T3 T4 Capacity (g / kg / day) 0 0 5 5 5 5 The number of animals 5 5 5 5 5 5 Womb 0.65 (0.18) ^** 0.10 ^a (0.02) ^b 0.10 (0.01) 0.10 (0.01) 0.23 ^* (0.04) 0.11 (0.02) lungs 1.24 (0.12) 1.31 (0.24) 1.45 (0.15) 1.52 (0.14) 1.32 (0.08) 1.40 (0.08) Heart 0.85 ^* (0.06) 1.05 (0.09) 1.10 (0.09) 1.09 (0.09) 0.87 ^* (0.08) 1.11 (0.16) liver 6.47 (0.63) 7.55 (0.9) 8.4 (0.61) 8.39 (1.42) 7.76 (0.48) 8.17 (0.66) spleen 0.40 ^** (0.05) 0.58 (0.07) 0.62 (0.10) 0.58 (0.1) 0.58 (0.09) 0.64 (0.06) Left kidney 0.77 (0.07) 0.79 (0.07) 0.89 (0.09) 0.87 (0.07) 0.80 (0.07) 0.92 (0.13) Right kidney 0.77 (0.05) 0.79 (0.08) 0.87 (0.06) 0.88 (0.04) 0.81 (0.08) 0.92 (0.13) Adrenal gland (left) 0.03 (0.01) 0.03 (0.02) 0.03 (0.0) 0.03 (0.01) 0.02 (0.0) 0.03 (0.0) Adrenal gland (right) 0.03 (0.0) 0.03 (0.01) 0.03 (0.01) 0.03 (0.01) 0.02 (0.01) 0.02 (0.01)

a; Numerical values are expressed as mean, b; ± S.D., Statistically significant difference with the negative control (* p <0.05, ** p <0.01).

<3-1-5> Abdominal fat measurement

At necropsy, the abdominal wall fat and abdominal fat of test animals were separated and weighed using an animal organ measuring scale.

As a result, the abdominal fat was increased by 264% in the negative control group from which the ovary was extracted (p <0.01). (FIG. 5 and Table 4).

gender female Experimental group C N.C T1 T2 T3 T4 Capacity (g / kg / day) 0 0 5 5 5 5 The number of animals 10 10 10 10 10 10 Abdominal wall fat (g) 3.83 ^** (1.85) 9.72 ^a (2.74) ^b 7.31 (1.67) 7.04 (1.87) 3.48 ^** (1.23) 8.68 (3.97) Abdominal fat (g) 6.7 ^** (1.76) 18.03 (7.36) 14.34 (4.6) 12.98 ^* (3.94) 5.05 ^** (2.55) 17.22 (3.31)

a; Numerical values are expressed as mean, b; ± S.D., Statistically significant difference with the negative control (* p <0.05, ** p <0.01).

<3-2> Hematological tests related to obesity and hyperlipidemia

<3-2-1> Hormone Concentration

Blood estrogen content was measured from blood collected at necropsy. As a result, the estrogen content was decreased by 21.37% in comparison with the normal control group. In the soy extract administration group (T1) and soybean fermentation administration group (T2), estrogen concentrations decreased by 4.49 and 7.62%, respectively, compared to the normal control group, and increased compared to the negative control group. In the fermented product group (T4), 100.46% and 117.65%, respectively, were significantly increased than the negative control group. In particular, the T4 group was statistically significant (p <0.01) increased (Table 5).

gender female Experimental group C N.C T1 T2 T3 T4 Capacity (g / kg / day) 0 0 5 5 5 5  14 week doses Estrogen (pg / ㎗) 183.38 ^a (44.37) ^b 144.2 (48.64) 175.14 (38.86) 169.4 (38.06) 184.22 (32.36) 215.75 ^** (42.88)

a; Numerical values are expressed as mean values, b; ± S.D., Statistical significance compared to negative control (** p <0.01).

<3-2-2> Quantitative Analysis of Total Cholesterol in Blood

Plasma cholesterol levels are indicative of coronary artery disease, thyroid function, liver function, bile secretion function, intestinal absorption and adrenal disease. Stress, aging, pregnancy and hormonal balance can affect cholesterol levels. Cholesterol esters in plasma are hydrolyzed to cholesterol and free fatty acids by cholesterol estase, and cholesterol is oxidized to cholesterol and 4-en-3-one and hydrogen peroxide by cholesterol oxidase. The hydrogen peroxide thus produced is combined with hydroxybenzoic acid and 4-aminoantipyrine in the reagent to develop color, and the amount of total cholesterol can be quantified by quantifying it at 510 nm / 620 nm.

In the present invention, Roeschlau's method (Roeschlau P, et al., Clin. Chem., 12: 226, modified from Allain's method (Allain CC, et al., Clin. Chem., 20: 470-475, 1974) 1974) to quantify total cholesterol. Specifically, three mice were orally administered for three days with one mouse, and physiological saline was orally administered to the control group. After fasting for 16 hours, Triton WR-1339 200 mg / kg was dissolved in sterile saline solution and injected into the tail vein, and normal group was injected with physiological saline instead of Triton WR-1339. Blood was collected. Samples were administered once daily for three days, but the final administration of the samples was made 1 hour prior to injection of Triton WR-1339. At this time, as a comparative group was used (Jenical (Roche Corporation)), the results are shown in Table 6.

<3-2-3> Quantification of HDL-Cholesterol in Blood

Aerobic exercise and eating foods high in vegetables, fruits and unsaturated fatty acids increase the amount of HDL-cholesterol in the blood. HDL-cholesterol is quantified by polyanion precipitation, in which lipoproteins, including positively charged apoprotein B, are combined with polyanions (Burstein M, et al. , J Lipid Res., 11: 583, 1970). PEG with an average molecular weight of 6,000 daltons can separate HDL-cholesterol by precipitating macromolecules such as lipoproteins at specific concentrations based on the size of the molecules or particles. The amount of HDL-cholesterol was quantified by the obtained supernatant. At this time, Xenical was used as a comparison group, and the results are shown in Table 6.

<3-2-4> LDL-cholesterol quantitative analysis

LDL-cholesterol is a factor that causes arteriosclerosis by accumulating on the blood vessel wall, and the amount of LDL-cholesterol in plasma is increased by ingestion of animal viscera, liver, eggs, red lean meat and saturated fatty acid. After separating serum from the blood, the present inventors separated the chylomicron by high-speed centrifugation at 13,000 rpm for 30 minutes, and separated the chylomicron at 2 ° C. using a density (1.063 g / ml) at 35,000 ° C. LDL-cholesterol was obtained by high-speed centrifugation for 24 hours at rpm. Subsequently, LDL-cholesterol was purified using a Cobas Mira autoanalyzer, Boehringer Mannheim GmbH Diagnostica, Mannheim, Germany, Kit No. 310328 and a Wako Chemical GmbH kit (Neuss, Germany, Kit No. 909-54009). Quantification At this time, Xenical was used as a comparison group, and the results are shown in Table 6.

<3-2-5> Quantitative Analysis of Triglycerides in Blood

Triglyceride (TG) is a type of blood lipid produced by the conversion of extra calories in the body and is associated with the development of obesity, hyperglycemia, heart disease and fatty liver. The inventors measured the amount of triglycerides using a specific chemistry analyzer (Kone, Finland) using an enzymatic colorimetric method (Boehringer Diagnostica, Mannheim, GmbH, FRG, kit No. 236691, 310328, 701912). At this time, Xenical was used as a comparison group, and the results are shown in Table 6.

As a result, in the case of the brown or brown fermentation group, the total cholesterol, LDL-cholesterol and triglyceride content all decreased significantly compared to the negative control group, and the HDL-cholesterol content tended to increase compared to the negative control group (Table 6 ).

Dose (mg / kg) Blood concentration (mg / dl) TC TG HDL LDL Normal control - 69.5 ± 5.2 48.5 ± 5.9 34.0 ± 1.1 26.5 ± 1.2 Negative Control - 193.6 ± 5.0 159.9 ± 10.43 26.8 ± 0.8 143.0 ± 0.9 L. Ashdophyllus 10 172.9 ± 11.2 147.2 ± 9.9 - - L. Bulgaricus 10 193.6 ± 5.0 159.9 ± 10.43 - - B. Breb K-111 10 153.2 ± 8.9 139.5 ± 9.2 - - B. Long Gum 10 183.5 ± 6.2 157.2 ± 9.5 - - S. Passion 10 173.3 ± 11.2 149.2 ± 7.8 - - L. Mesenteroides 10 183.5 ± 4.0 136.5 ± 11.2 - - Root Extract 100 129.1 ± 17.4 118.0 ± 18.6 29.3 ± 0.8 87.2 ± 2.4 Brown water extract 100 137.0 ± 4.5 105.8 ± 8.5 30.0 ± 0.5 95.7 ± 3.0 Brown root fermented product example <1-3> 100 (lactic acid bacteria 10 mg) 112.1 ± 15.3 105.0 ± 16.2 28.2 ± 1.7 85.8 ± 11.2 Brown root fermentation example <1-4> 100 (lactic acid bacteria <1 mg) 92.1 ± 15.3 75.0 ± 16.2 29.5 ± 1.5 65.8 ± 7.1 Fermented Fermentation (1) 100 (lactic acid bacteria 10 mg) 98.5 ± 17.8 88.3 ± 12.4 31.2 ± 3.2 81.2 ± 9.2 Fermented Fermentation (2) 100 (lactic acid bacteria <1 mg) 138.5 ± 7.8 102.3 ± 7.4 32.1 ± 1.5 91.8 ± 12.1 Zenical 10 87.6 ± 3.3 67.6 ± 6.8 31.5 ± 2.3 64.5 ± 2.2

Preparation Example 1 Preparation of a Pharmaceutical Composition Containing Brown Root or Browning Fermentation

<1-1> Preparation of Syrup

A syrup containing 2% (weight / volume) of an active ingredient of brown root or browned fermentation was prepared by the following method. First, the brown root or browned fermented powder, saccharin, and sugar prepared in Example 1 were dissolved in 80 g of warm water. After the solution was cooled, a solution consisting of glycerin, saccharin, spices, ethanol, sorbic acid and distilled water was prepared and mixed thereto. Water was added to this mixture to 100 ml.

The components of the syrup are as follows.

Brown or brown fermentation --------------------------------- 2 g

Saccharin ---------------------------------------------- 0.8 g

Party ------------------------------------------------- 25.4 g

Glycerin -------------------------------------------- 8.0 g

Spice --------------------------------------------- 0.04 g

Ethanol ---------------------------------------------- 4.0 g

Sorbate -------------------------------------------- 0.4 g

Distilled Water ----------------------------------------------- Quantification

<1-2> Preparation of Tablet

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

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

The components of the tablet are as follows.

Brown or brown fermentation ------------------------- 250 g

Lactose ------------------------------------ 175.9 g

Potato Starch -------------------------------------- 180 g

Colloidal Silicate -------------------------------- 32 g

10% gelatin solution

Potato Starch -------------------------------------- 160 g

Talc ------------------------------------------- 50 g

Magnesium Stearate ----------------------------- 5 g

<1-3> Preparation of Injection Solution

Injectable solutions containing 10 mg of the active ingredient were prepared in the following manner.

1 g of brown or brown fermentation, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

The components of the injection solution are as follows.

Brown or brown fermentation product --------------------------- 1 g

Sodium Chloride ------------------------------------ 0.6 g

Ascorbic acid ---------------------------------- 0.1 g

Distilled Water ----------------------------------------- Quantitative

Preparation Example 2 Preparation of Health Food Containing Brown Root or Brown Fermentation

<2-1> Preparation of food

Foods containing brown or brown fermentation were prepared as follows.

1. Preparation of Cooking Seasonings

Health promotion cooking seasonings were prepared from 20-95% by weight of brown or brown fermented products.

2. Preparation of Tomato Ketchup and Sauce

Health care tomato ketchup or sauce was prepared by adding 0.2 to 1.0% by weight of brown or brown fermented fermentation to tomato ketchup or sauce.

3. Manufacturing of Flour Foods

0.5 to 5.0% by weight of the brown or fermented fermentation product was added to the flour, and the mixture was used to prepare bread, cake, cookies, crackers, and noodles to prepare health promoting food.

4. Preparation of soups and gravy

0.1-5.0% by weight of brown or brown fermented fermentation was added to the soup and gravy to prepare a health promoting meat product, a soup of noodles and a gravy.

5. Preparation of Ground Beef

Health supplemented ground beef was prepared by adding 10% by weight of brown root or ground fermented product to ground beef.

6. Manufacture of Dairy Products

5-10% by weight of the brown or brown fermentation product was added to the milk, and the milk was used to prepare various dairy products such as butter and ice cream.

7. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh. The dried root or browned fermented product prepared in Example was decompressed and concentrated in a vacuum concentrator, and the dried product obtained by drying with spray and hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder. The grains, seeds, and dry powder of brown or fermented fermentation prepared above were blended in the following ratio.

Cereals (30% by weight brown rice, 15% by weight barley, 20% by weight barley),

Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),

Dry powder (3% by weight) of brown or brown fermentation products,

Ganoderma lucidum (0.5% by weight),

Sulfur (0.5 wt%)

<2-2> Preparation of Drink

1. Preparation of Carbonated Drinks

5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make a syrup, and the syrup is 85-98. After sterilization at 20 ° C. for 180 seconds, the mixture was mixed with cooling water at a ratio of 1: 4, and 0.5 to 0.82% of carbon dioxide was injected to prepare a carbonated beverage containing brown or brown fermentation product.

2. Manufacture of health drinks

Immediately sterilize by homogeneously mixing subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%), and brown or brown fermented products. Healthy drinks were prepared by packing them in small packaging containers such as glass bottles and PET bottles.

3. Preparation of Vegetable Juice

5 g of brown or brown fermented product was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

4. Preparation of Fruit Juice

1 g of brown or brown fermentation was added to 1,000 ml of apple or grape juice to prepare a fruit juice for health promotion.

As discussed above, the brown or brown fermented products show an excellent obesity or hyperlipidemia prevention or treatment effect that cannot be obtained only by the brown or brown extract, and are fermented after extracting from water compared to conventional extracts extracted with alcohol, especially toxic methanol aqueous solution. Therefore, there is an advantage that does not require an additional advanced purification process for application to the human body.

Claims (10)

For the prevention or treatment of obesity or hyperlipidemia containing lactic acid bacteria fermented product of the root or brown water extract as an active ingredient, which is prepared by adding hot water to the root or brown water and inoculating the water extract with fermentation of lactic acid bacteria. Pharmaceutical compositions. The pharmaceutical composition of claim 1, wherein the lactic acid bacterium is selected from the group consisting of Bifidobacterium, Lactobacillus, Streptococcus, Leukonostock, Pediococcus and Lactococcus microorganisms. The pharmaceutical composition according to claim 2, wherein the lactic acid bacteria is a microorganism in the genus Bifidobacterium. The pharmaceutical composition according to any one of claims 1 to 3, wherein the lactic acid bacteria are inoculated in an amount of 1 to 5% by weight of the sample weight. The pharmaceutical composition according to any one of claims 1 to 3, wherein the lactic acid bacteria are fermented at a temperature of 30 to 40 ° C for 24 to 52 hours. For the prevention or improvement of obesity or hyperlipidemia containing lactic acid bacteria fermented product of the root or brown water extract as an active ingredient, which is prepared by adding hot water to the root or brown water and inoculating the water extract by inoculating the lactic acid bacteria. Health food. The health food according to claim 6, wherein the lactic acid bacteria is selected from the group consisting of Bifidobacterium, Lactobacillus, Streptococcus, Leukonostock, Pediococcus and Lactococcus microorganisms. The health food according to claim 7, wherein the lactic acid bacteria is a microorganism in the genus Bifidobacterium. The health food according to any one of claims 6 to 8, wherein the lactic acid bacteria are inoculated in an amount of 1 to 5% by weight of the sample weight. The health food according to any one of claims 6 to 8, wherein the lactic acid bacteria are fermented at a temperature of 30 to 40 ° C for 24 to 52 hours.

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