KR101271478B1 - Composition for promoting leptin secretion comprising 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine as effective component - Google Patents

Abstract

The present invention is an appetite suppressant containing one or more components selected from the group consisting of 3-hydroxyflavone, 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine Leptin secretion-promoting composition and one or more components selected from the group consisting of 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine It relates to a functional food for promoting appetite suppressing hormone leptin (leptin) secretion containing as an active ingredient.

Description

Composition for promoting leptin secretion comprising 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine as effective component}

The present invention is an appetite suppressant containing one or more components selected from the group consisting of 3-hydroxyflavone, 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine It relates to a composition for promoting the secretion of leptin which is a hormone and a functional food for promoting the secretion of leptin.

Obesity is the highest in the United States (32% of the population), with a body mass index (BMI) of 30 or higher, and Mexico, 30%, the UK, 23%, Greece, Australia, and New Zealand, 21%. have. In Korea, 1.5% to 2 million people, 3.5% of the total population, are obese, but the population of obesity is increasing very rapidly due to westernization and diversification of dietary life.

Obesity has led to adult disease, including diabetes, hypertension, cardiovascular disease, cancer and mental distress, to some of the causes of death, and also affects national productivity, activity and competitiveness. In particular, the obesity of children and adolescents in Korea, which has undergone many changes in their diet and educational environment, is increasing rapidly, and the recovery of treatment after the onset of pediatric diabetes, etc., is not easy.

Obesity-related treatments have been tried in various ways such as drug administration, exercise, diet, psychotherapy, and surgery, but it is difficult to improve by specific methods, and complex treatments and patient will are important. In the case of drug administration, the drug was developed on the basis of lipolysis or digestive enzyme inhibition, but it has side effects.

As a solution to obesity, a method of degrading and accumulating mast cells or inhibiting digestive enzymes has been generally attempted. If food is supplied to the body, digestion or lipolysis has a limited role compared to the rate and amount of ingestion.

Thus, appetite control (appetite control) to fundamentally regulate the intake of food can be an approach to the fundamental solution, the appetite suppression by the food ingredient composition as a first attempt is meaningful as an invention with novelty, originality and progressiveness Have

Currently, reductil has been released as a drug to control obesity due to appetite control, but it is required to access food or natural products with side effects such as loss of appetite, insomnia, depression, and blood pressure. Therefore, there is a need for a composition for appetite suppression that can contribute to suppression of obesity by creating a new value as an appetite suppressing food material while minimizing side effects.

The present invention first developed an appetite suppression method by a food ingredient composition as a solution to obesity suppression, and in the method of proving appetite suppression effect, a test system that promotes the secretion of leptin that suppresses appetite for the first time to discover cells According to the degree of differentiation, food ingredients were applied and appetite suppression effect was scientifically verified.

Korean Patent Publication No. 2004-0097813 discloses an anti-obesity functional composition having three functions such as appetite suppression, body fat reduction, and bowel activation. Korean Patent Publication No. 2004-0036111 discloses ginseng and polyphenol-based substances or bioflavonoids. Foods for improving lipid metabolism and anti-obesity including powders or extracts of plants containing systemic substances have been disclosed, but leptin secretion comprising 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine of the present invention as active ingredients It is different from the composition for promotion.

The present invention is derived from the above requirements, the present invention is desired to develop a functional material that can suppress the appetite, but many limitations due to safety problems such as limitations and side effects of body fat decomposition, digestion inhibition method Nowadays, at least one selected from the group consisting of 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine, which are food ingredients and can suppress appetite The present invention was completed by developing a composition for promoting leptin secretion, which is an appetite suppressing hormone containing an ingredient as an active ingredient.

In addition, the present invention first developed an appetite suppression method by a food ingredient composition as a solution to obesity suppression, and also for the first time to discover a test system that promotes the secretion of leptin to suppress appetite in the method of proving appetite suppression effect According to the degree of cell differentiation, food ingredients were applied and appetite suppression effect was scientifically verified.

In order to solve the above problems, the present invention is one selected from the group consisting of 3-hydroxyflavone, 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine It provides a composition for promoting leptin secretion that is an appetite suppressing hormone containing the above components.

In addition, the present invention contains an active ingredient containing at least one component selected from the group consisting of 3-hydroxyflavone, hesperidin, hesperidin, quercetin, catechin and caffeine It provides a functional food for promoting the secretion of leptin, an appetite suppressing hormone. In the method of proving appetite suppression effect, a test system that promotes leptin, which suppresses appetite, was first discovered, and the food component was applied according to the degree of cell differentiation, and the appetite suppression effect of the food component was scientifically verified.

According to the present invention, the composition comprising 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine of the present invention as an active ingredient is leptin It is effective in suppressing appetite by promoting secretion, and since it uses food ingredients, no side effects appear even after long-term use, and safety can be ensured, and thus, it will be a very useful invention in functional food industry for appetite suppression.

In addition, in the method of proving appetite suppression effect, a leptin test system that suppresses appetite was first discovered and a food model was applied according to the degree of cell differentiation to provide a model for scientifically verifying the appetite suppression effect.

Figure 1 shows the concentration of leptin (leptin) compared to the control when 3-hydroxyflavone (3-hydroxyflavone) is added to the concentration of 0.01 ~ 10 ㎍ / ㎖ to 3T3-L1 cells, respectively.
Figure 2 is a measure of the concentration of leptin (leptin) compared to the control when hesperidin (hesperidin) is added to the concentration of 0.01 ~ 10 ㎍ / ㎖ to 3T3-L1 cells, respectively.
Figure 3 shows the concentration of leptin (leptin) compared to the control when quercetin (quercetin) is added to the concentration of 0.01 ~ 10 ㎍ / ㎖ to 3T3-L1 cells, respectively.
Figure 4 is a measure of the concentration of leptin (leptin) compared to the control when the catechin (catechin) at a concentration of 0.01 ~ 10 ㎍ / ㎖ to 3T3-L1 cells, respectively.
Figure 5 is a measure of the concentration of leptin (leptin) compared to the control when added to caffeine (caffeine) at a concentration of 0.01 ~ 10 ㎍ / ㎖ to 3T3-L1 cells.
Figure 6 shows the weight change and dietary intake of the rats in the normal, high fat diet (control), high fat diet + 3-hydroxyflavone (3-hydroxyflavone) administration group (A) respectively .
Figure 7 shows the rats in the normal diet group (normal), high fat diet (control), high fat diet + 3-hydroxyflavone (3-hydroxyflavone) administration group (A), respectively, showing the activity of the rat.

In order to achieve the object of the present invention, the present invention is selected from the group consisting of 3-hydroxyflavone, 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine It provides a composition for promoting leptin secretion which is an appetite suppressing hormone containing one or more components.

Since the composition of the present invention promotes leptin secretion, which is an appetite suppressing hormone, it can be used for appetite suppression.

In the composition of the present invention, the optimum concentration of each of the 3-hydroxyflavone (3-hydroxyflavone), hesperidin (hesperidin) and catechin (catechin) is characterized in that 0.1 ~ 10 ug / ml, the 3-hydroxyflavone The optimal concentration is preferably 1 ug / ml, the optimal concentration of hesperidin is preferably 0.1 ug / ml, and the optimal concentration of catechin is 0.1-1 ug / ml. In addition, the optimal concentration of quercetin (quercetin) is characterized in that 5 ~ 15 ug / ml, preferably 10 ug / ml, the optimal concentration of the caffeine (caffeine) is 0.01 ~ 10 ug / ml, preferably Is 0.1 ug / ml.

The composition of the present invention may comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical dosage forms of the compositions of the present invention may also be used in the form of their pharmaceutically acceptable salts and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable set.

The composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., oral preparations, suppositories and sterilized injection solutions according to a conventional method have. Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl And various compounds or mixtures including cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. 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 formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, 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. . 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.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the composition of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg. 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.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention also contains, as an active ingredient, at least one component selected from the group consisting of 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine. It provides a functional food for promoting the secretion of leptin, an appetite suppressing hormone.

When the composition of the present invention is used as a food additive, the composition may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, in the manufacture of food or beverages the components of the invention are added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight relative to the raw materials. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the 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 foods in a conventional sense.

The functional beverage composition containing the composition of the present invention may contain various flavors, natural carbohydrates, and the like as additional components, as in general beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio 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 the composition of the present invention.

In addition to the above, the composition of the present invention includes 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, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, the composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

1. Securing Sample

(1) Selection of appetite suppression material

The food ingredient for appetite suppression test system is a single ingredient, and it is possible to anti-obesity, 3-hydroxyflavone, hesperidin, quercetin, catechin and caffeine. (caffeine) was obtained.

(2) Preparation of Cell Test Sample Solution

The target food ingredients were dissolved in DMSO, diluted and used for each concentration. The final concentration of DMSO was 0.1% or less to prepare a cell test sample solution.

2. Appetite Suppression Hormones Leptin  Secretion test

(1) 3T3-L1 cell culture

3T3-L1 cells, a cell line derived from mouse embryos, were used by Korea Cell Line Bank (KCLB, Korea), and were used at 100 ° C. in DMEM containing 10% FBS and 1% PS at 37 ° C and 5%. The cells were cultured in an incubator maintained at% CO ₂ . When the cells were confluent, the cells were passaged by treatment with trypsin-EDTA, and the medium was exchanged every 2 to 3 days. When the cells were 80% confluent, 0.5 mM methylisobutylxanthine, 0.25 uM dexamethasone and 10 ug / ml insulin were used to observe the differentiation of 3T3-L1 cells by appetite suppressant. differentiation was induced by exchanging with 10% FBS, 1% PS-DMEM containing), and then replaced with a culture solution containing 10 ug / ml insulin every other day. That is, when 3T3-L1 cells were confluent, they were treated with 0.5 mM methyl isobutyl xanthine, 0.25 uM dexamethasone, and 10 ug / ml insulin to induce differentiation for 10 days as a control. In order to see the results of the appetite suppression material treatment, the different concentrations of samples were treated and compared for 48 hours incubation. After the end of the experiment, the culture, cell lysate and triglyceride samples were taken and stored at −80 ° C. until used in the experiment.

(2) Leptin assay

After the end of the experiment, the recovered culture was centrifuged (3,000 rpm, 20 min, 4 ° C) to recover only the supernatant, and leptin was measured by human Leptin kit biosource (invitrogen). Leptin is measured as follows. That is, 100 ul of standard diluent buffer and 100 ul of sample were put in a well, and 100 ul of a biotinylated anti-human leptin (Biotin Conjugate) solution was added. After 2 hours of incubation at room temperature, the culture solution was completely removed and washed 4 times with diluted wash solution. After adding 100 ul of Streptavidin-HRP working solution, washing was repeated for 4 minutes at room temperature for 30 minutes. After adding 100 ul of stabilized chromogen and reacting for 30 minutes in the dark at room temperature, 100 ul of stop solution was added and the absorbance was measured at 450 nm within 2 hours.

3. Animal experiments

(1) Experimental animal breeding

The experimental animals were 4 weeks old C57BL / 6J male mice, and the day and night cycle (12 hours light / 12 hours night) was controlled and the indoor temperature was raised in the animal breeding room of 22 ~ 25 ℃. One week of adaptation mice were fed high fat diets supplemented with 40% bee tallow to normal diet for 8 weeks to induce the high fat diet group. During the experiment, the experimental animals were divided into the normal group and the high-fat diet group, and the dietary intake and body weight were measured periodically.

Experimental dietary composition (g / Kg diet) ingredient Normal diet ^{1 )} High fat diet ^{2 )} Casein 200 200 DL-Methionine 3 3 Corn starch 150 150 Sucrose 500 345 Cellulose 50 50 Corn oil 50 - Beer tallow - 205 Salt mixture 35 35 Vitamin mixture 10 10 Choline bitartrate 2 2 Fat% (Calories) 11.7 40.0

¹⁾ Normal diet: AIN-76A diet # 100000 (Dyets Inc., Bethlehem, PA, USA)

²⁾ High-fat diet: AIN-76 diet # 100496 (Dyets Inc., Bethlehem, PA, USA)

(2) Selection of candidate material for preliminary animal experiment

In the laboratory, 3-hydroxyflavone, a food ingredient expected to have anti-obesity effect, was treated by intraperitoneal administration to high-fat diet mice at a concentration of 10 mg / kg for 3 weeks.

(3) measuring locomotor activity

Use the S-mart program (Pan Lab, Spain), a program that measures all movements of a given place using a computer sensor, for 1 hour in a white matt acrylic box of 40 × 40 × 40 cm in width, length, and height. The behavior of was measured after 3 weeks of food ingredient treatment. The image obtained from the digital camera installed about 2.5m above the box is transferred to the computer and tracked according to the movement of the subject by recognizing the center point of the white subject image using the principle of contrasting the white subject against the black background. It was. Then, the trajectory of the movement of the test animal was quantified to quantify the distance moved.

Example  One: When adding food ingredients Leptin  Concentration change

When 3T3-L1 adipocytes are differentiated, leptin is produced and secreted into cells. The secretion of leptin in the culture medium after sample treatment was 0.1 to 10 ug / ml of caffeine, and 3-hydroxyflavone, hesperidin and catechin at 0.1 to 10 ug / ml compared to the control group. In the concentration of ug / ml ~ 10 ug / ml, quercetin (quercetin) showed a higher leptin content than the control at a concentration of 10 ug / ml was found to be effective in suppressing appetite. Specifically, 3-hydroxyflavone showed the highest concentration of leptin at 1 ug / ml with a 39% increase compared to the control, and 7-8 at 0.1 ug / ml and 10 ug / ml. % Increased. Increasing leptin concentrations resulted in a 13% increase in quercetin at 10 ug / ml compared to the control, while hesperidin was 25% at 0.1 ug / ml and 8 at 1-10 ug / ml. % Increased. In addition, catechin was increased by 10% compared to the control at 0.1 ~ 1 ug / ml, caffeine (15% at 0.01 ug / ml, 27% at 0.1 ug / ml, 1 ~ 10 ug compared to the control) 7-8% increase in / ml.

The increased leptin content in the cell culture was synthesized in adipocytes, and the increase in leptin was estimated to be the primary increase factor. In addition, these components promote the secretion of leptin (leptin) was determined to have an appetite suppression effect along with signaling in the hypothalamus (Figs. 1 to 5).

Example  2: Changes in Dietary Intake and Weight of Laboratory Animals

As a result of comparing the weight change according to the normal diet group, the high fat diet group, and the high fat diet + 3-hydroxyflavone administration group, the weight gain was increased by 8-week high fat diet. Significantly increased, and when administered to 3-hydroxyflavone (3-hydroxyflavone), a decrease in dietary intake was shown, resulting in a decrease in weight gain (Fig. 6).

Example  3: amount of gait activity locomotor activity ) Measure

After 3 weeks of 3-hydroxyflavone administration to the high-fat diet group to determine whether the inhibition of weight gain following 3-hydroxyflavone administration was due to increased energy consumption In addition, the amount of walking activity was measured. After 1 hour of spatial adaptation, there was no significant difference in any group in the measurement of movement for 2 hours. Therefore, it was confirmed that the effect of increasing the weight increase by the administration of 3-hydroxyflavone (3-hydroxyflavone) was due to a decrease in dietary intake (Fig. 7).

Claims (5)

Promotes the secretion of leptin, an appetite suppressing hormone containing at least one component selected from the group consisting of 3-hydroxyflavone, hesperidin, quercetin and caffeine as an active ingredient. Food composition. According to claim 1, wherein the concentration of each of the 3-hydroxyflavone (3-hydroxyflavone) and hesperidin (hesperidin) is a composition for food, characterized in that 0.1 ~ 10 ug / ml. According to claim 1, wherein the concentration of quercetin (quercetin) is a composition for food, characterized in that 5 ~ 15 ug / ml. According to claim 1, wherein the concentration of caffeine (caffeine) is a composition for food, characterized in that 0.01 ~ 10 ug / ml. delete

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