KR102599530B1 - A composition for weight control comprising benzopyran - Google Patents

Abstract

The present invention relates to a composition for weight loss, and by containing a compound represented by the following [Chemical Formula 1] or an acceptable salt thereof as an active ingredient, it is possible to not only control body weight but also improve, prevent or treat obesity. Additionally, since it is non-toxic, it can be consumed in food form.
[Formula 1]

Description

A composition for weight loss comprising a benzopyrene derivative as an active ingredient {A composition for weight control comprising benzopyran}

The present invention relates to a composition capable of reducing body weight by containing a benzopyrene derivative as an active ingredient.

Westernized eating habits and adults' failure to control their weight lead to a rapid increase in obesity rates and cause many negative consequences for our health and quality of life. As metabolic diseases caused by obesity gradually increase, social costs increase. It's a trend.

Obesity can be defined as a type of disease that poses a threat to an individual's health due to abnormally excessive fat accumulation in the body. Obesity is divided into simple obesity, which is mainly caused by overeating and lack of exercise, and symptomatic obesity, which is caused by endocrine diseases.

Causes of simple obesity include incorrect eating habits such as overeating, binge eating, snacking, late-night snacking, and irregular eating, lack of exercise, and side effects of medications.

There have been many recent changes in eating habits due to economic growth and changes in lifestyle. In particular, busy modern people are increasingly overweight and obese due to high-calorie diets such as fast food and little exercise. According to the World Health Organization (WHO), more than 1 billion adults worldwide are overweight, of which at least 3 million are clinically obese, especially in Europe, where 250,000 people are overweight each year, and more than 2.5 million worldwide. There have been reports of deaths related to being overweight.

Overweight and obesity increase blood pressure and cholesterol levels, which can cause or worsen various chronic complications such as heart disease, diabetes, arthritis, fatty liver, hyperlipidemia, and cancer. Additionally, overweight and obesity are known to be major factors that increase the incidence of arteriosclerosis, high blood pressure, hyperlipidemia, or heart disease not only in adults but also in children and adolescents.

Therefore, the need to recognize obesity as a disease and actively treat it is increasing.

To date, the substances used to treat obesity are drugs that reduce fat absorption in the stomach or suppress appetite. However, Xenical (Roche) and Alli (GlaxoSmithKline), which work to prevent fat absorption by inhibiting lipase in the small intestine and pancreas, have been reported to cause side effects such as gastrointestinal side effects and reduced absorption of fat-soluble vitamins.

In addition, lorcaserin (5-HT2c receptor agonist) and Qnexa (Phentermine/Topiramate), which suppress obesity by suppressing appetite, may cause side effects such as attention deficit or memory impairment, and when taken together with antidepressants or migraine drugs, It has been reported that it has the potential to cause serious side effects such as heart valve disease, and sibutramine (serotin noradrenalin reuptake inhibitor) was withdrawn in 2010 due to an increase in the incidence of cardiovascular disease and cerebral infarction.

Republic of Korea Patent Publication No. 2019-0022085 Republic of Korea Registered Patent No. 1761155

The purpose of the present invention is to provide a food composition for weight loss containing a benzopyrene derivative as an active ingredient.

In addition, another object of the present invention is to provide a food composition for improving or preventing obesity containing a benzopyrene derivative as an active ingredient.

In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity containing a benzopyrene derivative as an active ingredient.

In addition, another object of the present invention is to provide a veterinary composition for weight loss in animals other than humans containing a benzopyrene derivative as an active ingredient.

In addition, another object of the present invention is to provide a feed additive for weight loss in animals other than humans containing a benzopyrene derivative as an active ingredient.

The food composition for weight loss of the present invention for achieving the above object may contain a benzopyrene derivative, such as a compound represented by the following [Formula 1] or an acceptable salt thereof as an active ingredient;

[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group,

R ₂ to R ₅ are a group consisting of a hydroxy group, C _1-8 alkyl group, cyano group, nitro group, amino group, amidino group, hydrazine, hydrazone, carboxyl group or salts thereof, sulfonic acid group or salts thereof, phosphoric acid or salts thereof are independently selected to be the same or different from each other.

In addition, the food composition for improving or preventing obesity of the present invention for achieving the other purposes described above may contain the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient.

The composition can inhibit the formation of lipid droplets and differentiation into adipocytes when inducing differentiation of preadipocytes into adipocytes.

The composition can inhibit the expression of lipid synthesis-related genes such as ACCα (acetyl CoA carboxylase α), ACLY (ATP-citrate synthase), and FAS (fatty acid synthase); It can suppress the expression of transcription-related genes caused by lipid accumulation of C/EBP-β, C/EBP-α, PPAR-γ1, PPAR-γ2, and LXRα.

In addition, the composition can inhibit the expression of genes related to neutral lipid (TG) synthesis, such as GPAT, AGPAT2, DGAT1, and MOGAT1; It can suppress the expression of lipid transport-related genes such as aP2/FABP4, FSP27, and CD36.

In addition, the pharmaceutical composition for preventing or treating obesity of the present invention to achieve the above-described other purpose may contain the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient.

In addition, the veterinary composition for weight loss in animals other than humans of the present invention to achieve the above-described other purpose may contain the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient.

In addition, the feed additive for weight loss in animals other than humans of the present invention to achieve the above-described other purpose may contain the compound represented by [Chemical Formula 1] or an acceptable salt thereof as an active ingredient.

The composition of the present invention has a significant effect in preventing or treating obesity because it reduces body weight and suppresses the expression of obesity-related genes.

Additionally, since the present invention has no side effects, it can be used long-term for weight loss and obesity.

Figure 1 is a graph measuring the body weight of the sample administration group, normal group, and control group mice administered 0.1 g/kg and 0.25 g/kg of tamaricetin of Example 1, respectively, for 10 weeks.
Figure 2 shows the control group, MDI group, and MDI+Example 1 (50 uM) stained with Oil Red O solution to confirm the inhibitory effect of tamaricetin on lipid droplet formation when inducing adipocyte differentiation of 3T3-L1 preadipocytes. and an absorbance graph measured after dissolving 3T3-L1 preadipocytes of MDI+Example 1 (100 uM).
Figure 3 shows the control group, MDI group, and MDI+Example 1 ( This is a view confirming the group treated with MDI + Example 1 (100 uM) under a microscope.
Figure 4a shows the FAS gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 4b shows the ACLY gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 4c shows the ACCα gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.
Figure 5a shows C/EBP-β observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). about genes It is a graph; Figure 5b shows C/EBP-α observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). about genes It is a graph; Figure 5c shows the PPAR-γ1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 5d shows the PPAR-γ2 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 5e shows the LXRα gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.
Figure 6a shows the GPAT gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6b shows the AGPAT2 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6c shows the DGAT1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6d shows the MOGAT1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.
Figure 7a shows the aP2/FABP4 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 7b shows the FSP27 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 7c shows the CD36 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.

The present invention relates to a composition capable of reducing body weight by containing a benzopyrene derivative as an active ingredient.

Tamaricetin or its acceptable salt, which is one of the benzopyrene derivatives, is only used as a composition for preventing or treating sepsis or septic shock, a composition for preventing or treating tuberculosis, etc. Tamaricetin or its acceptable salt is used for weight loss. It has never been used as a composition.

Hereinafter, the present invention will be described in detail.

The composition for weight loss of the present invention contains a compound represented by the following [Formula 1] or an acceptable salt thereof as an active ingredient.

[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group,

R ₂ to R ₅ are a group consisting of a hydroxy group, C _1-8 alkyl group, cyano group, nitro group, amino group, amidino group, hydrazine, hydrazone, carboxyl group or salts thereof, sulfonic acid group or salts thereof, phosphoric acid or salts thereof are independently selected to be the same or different from each other.

In addition, the present invention can be used as a composition that can improve, prevent, or treat obesity by containing the compound represented by [Chemical Formula 1] or an acceptable salt thereof as an active ingredient.

The composition that can improve, prevent or treat obesity inhibits lipid accumulation and inhibits the expression of ACCα (acetyl CoA carboxylase α), ACLY (ATP-citrate synthase) and FAS (fatty acid synthase), which are representative genes related to intracellular lipid synthesis. It inhibits the activity of transcription-related genes C/EBP-β, C/EBP-α, PPAR-γ1, PPAR-γ2, and LXRα due to lipid accumulation, as well as genes related to neutral lipid (TG) synthesis. It was confirmed that it inhibits the activities of GPAT, AGPAT2, DGAT1, and MOGAT1. In addition, it was confirmed that the activities of lipid transport-related genes aP2/FABP4, FSP27, and CD36 were also inhibited.

Meanwhile, in this specification, the term ‘containing as an active ingredient’ means containing a sufficient amount to achieve the efficacy or activity of the compound represented by [Formula 1] or an acceptable salt thereof. For example, the compound represented by [Formula 1] or an acceptable salt thereof is used at a concentration of 10 to 1500 μg/ml, preferably 50 to 1000 μg/ml. Since the compound represented by [Formula 1] or an acceptable salt thereof has no side effects on the human body, the upper quantitative limit of the compound represented by [Formula 1] or an acceptable salt thereof contained in the composition of the present invention can be selected within an appropriate range by a person skilled in the art. It can be implemented.

The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable auxiliaries in addition to the active ingredients, and the auxiliaries include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, Lubricants or flavoring agents can be used.

For administration, the pharmaceutical composition may be preferably formulated as a pharmaceutical composition containing one or more pharmaceutically acceptable carriers in addition to the active ingredients described above.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, etc. Additionally, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacance or sodium oleate, sodium stearate, magnesium stearate, sodium Includes benzoate, sodium acetate, sodium chloride, etc. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, etc.

Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these ingredients can be mixed and used, and other common additives such as antioxidants, buffers, and bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Furthermore, it can be preferably formulated according to each disease or ingredient using a method disclosed by Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, as an appropriate method in the field.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., and is preferably administered orally. .

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. Usually, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention can be prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or can be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, or capsule, and may additionally contain a dispersant or stabilizer.

In addition, the present invention provides a food composition for weight loss or for improving, preventing or treating obesity, containing the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient.

The food composition according to the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, and health supplements. etc.

The food composition of the present invention may contain not only the compound represented by [Chemical Formula 1] or an acceptable salt thereof as an active ingredient, but also ingredients commonly added during food production, such as proteins, carbohydrates, fats, Contains nutrients, seasonings and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured as a drink or beverage, in addition to the compound represented by [Formula 1] of the present invention, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts are added. It can be included as .

The present invention provides a health functional food comprising a food composition for weight loss or for improving, preventing or treating obesity, containing the compound represented by the above [Formula 1] or an acceptable salt thereof as an active ingredient. Health functional foods are foods manufactured by adding a compound represented by [Chemical Formula 1] or an acceptable salt thereof to food ingredients such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending them. It means that it brings about specific health effects when ingested, but unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of the compound represented by [Chemical Formula 1] or its allowable salt in such health functional foods cannot be uniformly defined as it varies depending on the type of health functional food being targeted, but within the range that does not damage the original taste of the food. It can be added and is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight, relative to the target food. In addition, in the case of health functional foods in the form of pills, granules, tablets, or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule, or beverage.

In addition, the present invention provides the use of the compound represented by [Chemical Formula 1] or an acceptable salt thereof for the production of medicine or food for weight loss or for the improvement, prevention or treatment of obesity. As described above, the compound represented by [Chemical Formula 1] or an acceptable salt thereof can be used for weight loss or for the improvement, prevention or treatment of obesity.

Additionally, the present invention provides a weight control method comprising administering an effective amount of the compound represented by [Formula 1] or an acceptable salt thereof to a mammal.

As used herein, the term “mammal” refers to a mammal, preferably a human, that is the subject of treatment, observation or experiment.

As used herein, the term “effective amount” means the amount of an active ingredient or pharmaceutical composition that is believed by a researcher, veterinarian, physician, or other clinician to induce a biological or medical response in a tissue system, animal, or human, which means that It includes amounts that induce relief of symptoms of a disease or disorder. The effective amount and frequency of administration of the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by a person skilled in the art, and can be determined based on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of dosage form, and the patient's age, weight, and general health. It can be adjusted according to various factors, including condition, gender and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs. In the prevention, treatment or improvement method of the present invention, for adults, the compound represented by [Formula 1] or an acceptable salt thereof is administered once or several times a day at a dose of 0.001 g/kg to 10 g/kg. It is preferable to administer.

In the treatment method of the present invention, the composition containing the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, It can be administered in the conventional manner via intraocular or intradermal routes.

In addition, the veterinary composition capable of reducing body weight in animals other than humans according to the present invention includes a compound represented by [Chemical Formula 1] or an acceptable salt thereof as an active ingredient.

The veterinary composition containing the compound represented by [Chemical Formula 1] or an acceptable salt thereof of the present invention may further include appropriate excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition containing the compound represented by [Formula 1] or an acceptable salt thereof of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, Maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate. Salt, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben and mineral oil.

The veterinary composition containing the compound represented by [Formula 1] or an acceptable salt thereof according to the present invention may further include fillers, anti-coagulants, lubricants, wetting agents, spices, emulsifiers, preservatives, etc. Veterinary compositions can be formulated using methods well known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to an animal, and the dosage form may be powders, granules, tablets, capsules, It may be in the form of suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, suppository, sterile injectable solution, sterile topical solution, etc.

The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, but can be administered in an amount of 0.1 to 100 mg/kg once or several times a day, and the dosage depends on the route of administration and the degree of disease. , may increase or decrease depending on gender, weight, age, etc. Accordingly, the above dosage does not limit the scope of the present invention in any way.

In addition, the present invention relates to a feed additive for weight loss in animals other than humans, comprising the compound represented by [Formula 1] or an acceptable salt thereof as an active ingredient.

The feed additive may be in the form of a highly concentrated liquid, powder or granule containing 20 to 90% by weight of the compound represented by [Formula 1] or an acceptable salt thereof.

The feed additive of the present invention is organic acids such as citric acid, humalic acid, adipic acid, lactic acid, and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polymerized phosphate), polyphenol, catechin, and alpha. -It may additionally contain one or more selected from natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid.

The animal feed additive containing the compound represented by [Formula 1] or an acceptable salt thereof of the present invention and the feed containing the same contain amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, anti-fungal enzymes, etc. as auxiliary ingredients. It can be used with substances such as digestion and absorption enhancers, growth promoters, and disease prevention agents.

The feed additive may be administered to animals alone or in combination with other feed additives in an edible carrier. In addition, the feed additives can be easily administered as a top dressing or by mixing them directly into animal feed, or separately from the feed, in a separate oral formulation, by injection or transderm, or in combination with other ingredients. Typically, single daily doses or divided daily doses may be used, as is well known in the art. When the feed additive is administered separately from animal feed, dosage forms of the extract can be prepared in immediate-release or sustained-release formulations by combining them with non-toxic pharmaceutically acceptable edible carriers, as is well known in the art. These edible carriers can be solid or liquid, such as corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the dosage form of the extract may be tablets, capsules, powders, turmeric or sugar-containing tablets or a top dressing in microdisperse form. If a liquid carrier is used, the dosage form may be a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. Additionally, the dosage forms may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, etc. Additionally, animal feed containing the compound represented by [Formula 1] or an acceptable salt thereof as a feed additive may be any protein-containing organic grain flour commonly used to meet the dietary needs of animals. These protein-containing flours typically consist primarily of corn, soybean flour, or corn/soybean flour mixes.

The above feed additives can be used by adding them to the animal feed by dipping, spraying, or mixing. The veterinary composition or feed additive of the present invention can be applied to the diet of companion animals.

Additionally, the present invention relates to a weight control method comprising administering a compound represented by [Formula 1] or an acceptable salt thereof to an entity other than a human.

Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are merely illustrative of the present invention, and it is clear to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the attached patent claims.

Example 1.

It is a composition containing tamaricetin as an active ingredient.

<Test example>

[Animal testing]

The animals were 7-week-old male ob/ob mouse models weighing 18-22 g that were used in the experiment after being adapted to the laboratory environment for one week. Solid feed and water were provided ad libitum. The laboratory was maintained at a constant temperature (22 ± 2 °C) and humidity (50 ± 10%) and a 12-hour day/night cycle.

Test Example 1. Weight measurement

The experimental animals were selected only from healthy animals with consistent weight changes and divided into normal group, control group, and sample administration group by random placement method, and 7 animals were used in each experimental group. For the normal animal model experiment, the normal group fed a normal diet, the control group fed a high fat-high sucrose diet, and the high fat-high sucrose diet and Example 1 were tested by concentration (0.1 g/kg and The samples administered at 0.25 g/kg were divided into groups.

Administration was by intravenous injection into the tail vein once a day for 10 consecutive weeks, and the body weight of the mice was measured for 10 weeks.

Figure 1 is a graph measuring the body weight of the sample administration group, normal group, and control group mice administered 0.1 g/kg and 0.25 g/kg of tamaricetin of Example 1, respectively, for 10 weeks.

As shown in Figure 1, the weight of the control group fed only a high-fat diet increased the most, and the sample administration group fed with a high-fat diet and tamaricetin of Example 1 gained less weight than the control group. confirmed.

In particular, it was confirmed that from the 9th week, among the administration groups, the body weight of Example 1 0.25 g/kg decreased more than that of Example 1 0.1 g/kg.

[Cell experiment]

cell culture

3T3-L1 preadipocytes, which are mouse fibroblasts, were provided by ATCC (American Type Culture Collection, Manassas, VA, USA). 3T3-L1 preadipocytes were cultured in 90% DMEM medium (Dulbecco's Modified Eagle's Medium, GKibco BRL, Grand Island, NY, USA), 10% bovine calf serum (BCS, Gibco BRL, Grand Island, NY, USA), and 1% penicillin and streptomycin (Gibco BRL, Grand Island, NY, USA) and cultured under 37°C and 5% CO₂ conditions. In order to eliminate overdensity due to proliferation of cells, the growth medium was replaced every 48 hours to maintain an appropriate number of cells.

cell differentiation

To differentiate 3T3-L1 preadipocytes into adipocytes, 3T3-L1 preadipocytes were cultured to a confluent state using growth medium containing 10% bovine serum and 1% penicillin and streptomycin. In a confluent state, the cells were further cultured for 2 days using differentiation medium containing 10% fetal bovine serum (FBS, Gibco BRL, Grand Island, NY, USA) and 1% penicillin and streptomycin.

Afterwards, it was replaced with MID differentiation medium containing 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.1 μM dexamethasone, and 10 μg/ml insulin. The cells were cultured for 2 days, and then every 2 days, the cells were replaced with differentiation medium containing 10 μg/ml insulin.

In addition, in order to analyze the degree of inhibition of differentiation of 3T3-L1 preadipocytes by tamaricetin, the medium was treated with an appropriate concentration of tamaricetin when exchanged for differentiation medium containing MDI and insulin. 3T3-L1 preadipocytes induced to differentiate as described above were used for various experimental analyses.

oil red

Oil Red O solution and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) were purchased from Sigma-Aldrich (USA).

The term "Oil Red O staining" in the present invention refers to a type of staining method that stains intracellular fat red using Oil Red O. In addition, the term "Oil Red O solution" in the present invention refers to a solution in which Oil Red O is dissolved in a solvent such as isopropyl alcohol, ethanol, methanol, propylene glycol, etc., and the solvent is preferably isopropyl alcohol. The amount of propyl alcohol (isopropyl alcohol) included and dissolved may vary depending on the type of solvent and dyeing conditions, but it is preferable to dissolve it to the point of saturation in consideration of economics while maximizing dyeing efficiency.

Oil Red O staining is a dye-binding method used to quantify total lipids. Due to its characteristic of binding specifically to lipids, it is used to analyze the frequency of fat produced in adipocytes. Therefore, in order to analyze the effect of tamaricetin on lipid droplets generated in 3T3-L1 adipocytes induced by differentiation, Oil Red O staining was performed.

Test example 2. Confirmation of inhibition of lipid accumulation_quantitative confirmation

Figure 2 shows the control group, MDI group, and MDI+Example 1 (50 uM) stained with Oil Red O solution to confirm the inhibitory effect of tamaricetin on lipid droplet formation when inducing adipocyte differentiation of 3T3-L1 preadipocytes. and an absorbance graph measured after dissolving 3T3-L1 preadipocytes of MDI+Example 1 (100 uM).

Specifically, the following test was performed to quantitatively confirm intracellular lipid accumulation in the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM).

The control and 3T3-L1 adipocytes treated with different concentrations of tamaricetin had the medium removed, washed with phosphate buffered saline (PBS), and fixed with 3.7% formalin for 1 hour. After fixation, cells were washed with 60% isopropanol, then treated with Oil Red O solution, and staining was performed for 20 minutes at room temperature. After staining, the Oil Red O solution was removed, washed three times with distilled water, and the stained cells were photographed using a microscope.

In addition, in order to quantitatively analyze the degree of inhibition of triglyceride production induced by tamaricetin, the Oil Red O solution stained in the cells was dissolved using 100% isopropanol, and then 200 μl was added to a 96 well plate. The absorbance was measured at 520 nm using a microplate reader.

As shown in Figure 2, preadipocytes differentiated by MDI were treated at different concentrations using tamaricetin of Example 1 and changes in lipid accumulation were measured. As a result, the amount of lipid accumulation in the MDI group increased compared to the control group. , it was confirmed that when the MDI group was treated with Example 1 at concentrations of 50 uM and 100 uM, respectively, the amount of lipid accumulation was suppressed compared to the MDI group.

This means that the tamaricetin of Example 1 shows an excellent lipid accumulation inhibition effect.

Test Example 3. Confirmation of inhibition of fat accumulation_Photo

The cells of the control group, MDI group, MDI + Example 1 (50 uM) and MDI + Example 1 (100 uM) stained according to the method of Test Example 1 above were washed with 1 ml of water and then analyzed under a red visible imaging microscope (Olympus, Tokyo). , Japan).

Figure 3 shows the control group, MDI group, and MDI+Example 1 ( This is a view confirming the group treated with MDI + Example 1 (100 uM) under a microscope.

As shown in Figure 3, it was confirmed that the MDI + Example 1 (50 uM) group and the MDI + Example 1 (100 uM) group had significantly lower intracellular lipid accumulation than the MDI group. In particular, it was confirmed that almost no lipid accumulation was observed in the MDI+Example 1 (100 uM) group.

These results mean that Example 1 of the present invention has an excellent lipid accumulation inhibition effect.

Test example 4. FAS, ACLY and ACCα Expression measurements

Figure 4a shows the FAS gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 4b shows the ACLY gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 4c shows the ACCα gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.

Specifically, genes related to lipid synthesis In order to confirm the decrease in lipid synthesis due to decreased expression, changes in the expression of FAS, ACLY , and ACCα genes, which are representative genes related to lipid synthesis, were performed in the following test.

3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM) and MDI + Example 1 (100 uM) were collected, RNA was extracted, and reverse transcriptase was used using 2 ug of the extracted RNA as a template. After synthesizing cDNA, using this as a template, changes in expression of FAS, ACLY , and ACCα genes were observed using real-time qPCR (quantitative polymerase chain reaction).

As shown in Figures 4A to 4C, it was confirmed that the expression of FAS, ACLY , and ACCα genes due to lipid accumulation in the group treated in Example 1 was suppressed compared to the MDI group. In particular, it was confirmed that the MDI + Example 1 (100 uM) group suppressed the expression of FAS, ACLY , and ACCα genes better than the MDI + Example 1 (50 uM) group.

Test example 5. C/EBP-β, C/EBP-α, PPAR-γ1, PPAR-γ2 and LXRα Expression measurements

Figure 5a shows C/EBP-β observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). about genes It is a graph; Figure 5b shows C/EBP-α observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). about genes It is a graph; Figure 5c shows the PPAR-γ1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 5d shows the PPAR-γ2 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 5e shows the LXRα gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.

Specifically, transcription-related genes due to lipid accumulation In order to confirm the decrease in lipid accumulation due to decreased expression, representative genes related to transcription by lipid accumulation, such as C/EBP-β, C/EBP-α, PPAR-γ1, PPAR-γ2, and LXRα, were examined. Changes in gene expression were performed in the same manner as Test Example 3 above.

As shown in Figures 5A to 5E, C/EBP-β, C/EBP-α, PPAR-γ2, and LXRα in the group treated with Example 1 It was confirmed that the expression of genes was suppressed compared to the MDI group. In particular, it was confirmed that the MDI+Example 1 (100 uM) group suppressed the expression of C/EBP-β, C/EBP-α, PPAR-γ2, and LXRα genes better than the MDI+Example 1 (50 uM) group.

On the other hand, it was confirmed that the expression of the PPAR-γ1 gene was not suppressed in the group treated in Example 1.

Test example 6. GPAT, AGPAT2, DGAT1 and MOGAT1 Expression measurements

Figure 6a shows the GPAT gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6b shows the AGPAT2 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6c shows the DGAT1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 6d shows the MOGAT1 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.

Specifically, genes related to neutral lipid (TG) synthesis GPAT, AGPAT2, DGAT1 and MOGAT1 by reduced expression Changes in expression of genes were conducted in the same manner as Test Example 3 above.

As shown in Figures 6A to 6D, GPAT, AGPAT2, DGAT1 and MOGAT1 in the group treated with Example 1 It was confirmed that the expression of genes was suppressed compared to the MDI group. In particular, it was confirmed that the MDI + Example 1 (100 uM) group suppressed the expression of AGPAT2, DGAT1, and MOGAT1 genes better than the MDI + Example 1 (50 uM) group.

Test example 7. aP2/FABP4, FSP27 and CD36 Expression measurements

Figure 7a shows the aP2/FABP4 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). About It is a graph; Figure 7b shows the FSP27 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It is a graph; Figure 7c shows the CD36 gene observed by PCR (quantitative polymerase chain reaction) in 3T3-L1 preadipocytes treated with the control group, MDI group, MDI + Example 1 (50 uM), and MDI + Example 1 (100 uM). It's a graph.

Specifically, lipid transport-related genes To confirm the decrease in expression, changes in the expression of aP2/FABP4, FSP27 , and CD36 genes were performed in the same manner as Test Example 3.

As shown in Figures 7A-7C, aP2/FABP4, FSP27 and CD36 in the group treated with Example 1 It was confirmed that the expression of genes was suppressed compared to the MDI group. In particular, it was confirmed that the MDI+Example 1 (100 uM) group suppressed the expression of aP2/FABP4, FSP27 , and CD36 genes better than the MDI+Example 1 (50 uM) group.

Test Example 8. Confirmation of cytotoxicity

In the MDI group (induced group), 3T3-L1 cells were inoculated into a 24-well plate, 0.5 mM MDI was added to low glucose DMEM medium supplemented with 1% BSA, and cultured for 24 hours at 37°C and 5% CO ₂ conditions. Next, add 10 ㎕ of 5 mg/㎖ MTT stock, remove the medium after 3 hours, check the cells that turned purple, dissolve them in DMSO, transfer 100 ㎕ to a 96-well plate, and measure the absorbance at 540 nm to confirm cytotoxicity. .

In addition, for each treatment group, the DMEM medium of the MDI group was treated with tamaricetin of Example 1, cultured for 24 hours at 37 ° C. and 5% CO ₂ , and then 10 ㎕ of 5 mg / ㎖ MTT stock was added and 3 After a period of time, the medium was removed to confirm that the cells had turned purple, then dissolved in DMSO, transferred to 96-well plates (100 ㎕), and cytotoxicity was confirmed by measuring the absorbance at 540 nm.

division control group MDI MDI+Example 1
(50 uM) MDI+Example 1
(100 uM) cell viability
(%) 100 95.7 98.9 98.1

As shown in Table 1 above, it was confirmed that tamaricetin of Example 1 of the present invention did not show toxicity.

Below, a formulation example of a composition containing the powder of the present invention is described, but the present invention is not intended to be limited, but merely explained in detail.

Formulation Example 1. Preparation of powder

Tamaricetin of Example 1 500 mg

100 mg lactose

Talc 10 mg

The above ingredients are mixed and filled into an airtight bubble to prepare a powder.

Formulation Example 2. Preparation of tablets

Tamaricetin 300 mg of Example 1

Corn starch 100 mg

100 mg lactose

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are manufactured by compressing them according to a typical tablet manufacturing method.

Formulation Example 3. Preparation of capsules

Tamaricetin 200 mg of Example 1

3 mg crystalline cellulose

Lactose 14.8 mg

Magnesium stearate 0.2 mg

Capsules are prepared by mixing the above ingredients and filling them into gelatin capsules according to a typical capsule manufacturing method.

Formulation Example 4. Preparation of injections

Tamaricetin 600 mg of Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to the usual manufacturing method for injections.

Formulation Example 5. Preparation of liquid formulation

4 g of tamaricetin of Example 1

10 g isomerized sugar

5 g mannitol

Proper amount of purified water

According to the usual liquid preparation method, add and dissolve each ingredient in purified water, add an appropriate amount of lemon flavor, mix the above ingredients, add purified water, adjust the total to 100g by adding purified water, and then fill it in a brown bottle and sterilize it. to produce a liquid.

Formulation Example 6. Preparation of granules

Tamaricetin 1,000 mg of Example 1

Vitamin mixture dosage

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Monobasic Potassium Phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above vitamin and mineral mixture is a mixture of components relatively suitable for granules in a preferred embodiment, but the mixing ratio may be modified arbitrarily. The above components are mixed according to a typical granule manufacturing method, and then the granules are mixed. It can be prepared and used to manufacture a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

Tamaricetin 1,000 mg of Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g plum concentrate

1 g taurine

Add purified water to make a total of 900 mL.

After mixing the above ingredients according to a typical health drink manufacturing method, stirring and heating at 85°C for about 1 hour, the resulting solution was filtered, placed in a sterilized 2 L container, sealed, sterilized, stored in the refrigerator, and then refrigerated. Used for manufacturing the functional beverage composition of the invention.

The composition ratio is a preferred embodiment of mixing ingredients relatively suitable for beverages of preference, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, country of demand, and intended use.

Claims (11)

A food composition for weight loss containing a compound represented by the following [Chemical Formula 1] or an acceptable salt thereof as an active ingredient;
[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group, and R ₂ to R ₅ are each independently a hydroxy group. A food composition for improving or preventing obesity containing a compound represented by the following [Formula 1] or an acceptable salt thereof as an active ingredient;
[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group, and R ₂ to R ₅ are each independently a hydroxy group. The food composition for improving or preventing obesity according to claim 2, wherein the composition inhibits the formation of lipid droplets and differentiation into adipocytes when inducing differentiation of preadipocytes into adipocytes. The food for improving or preventing obesity according to claim 2, wherein the composition inhibits lipid synthesis-related genes of ACCα (acetyl CoA carboxylase α), ACLY (ATP-citrate synthase), and FAS (fatty acid synthase). Composition. The method of claim 2, wherein the composition improves or prevents obesity by suppressing transcription-related genes caused by lipid accumulation of C/EBP-β, C/EBP-α, PPAR-γ1, PPAR-γ2, and LXRα. For food composition. The food composition for improving or preventing obesity according to claim 2, wherein the composition inhibits genes related to triglyceride (TG) synthesis of GPAT, AGPAT2, DGAT1, and MOGAT1. The food composition for improving or preventing obesity according to claim 2, wherein the composition inhibits lipid transport-related genes of aP2/FABP4, FSP27, and CD36. A pharmaceutical composition for preventing or treating obesity containing a compound represented by the following [Chemical Formula 1] or an acceptable salt thereof as an active ingredient;
[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group, and R ₂ to R ₅ are each independently a hydroxy group. The pharmaceutical composition for preventing or treating obesity according to claim 8, wherein the composition inhibits the formation of lipid droplets and differentiation into adipocytes when inducing differentiation of preadipocytes into adipocytes. A veterinary composition for weight loss in animals other than humans containing a compound represented by the following [Formula 1] or an acceptable salt thereof as an active ingredient;
[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group, and R ₂ to R ₅ are each independently a hydroxy group. A feed additive for weight loss in animals other than humans containing a compound represented by the following [Chemical Formula 1] or an acceptable salt thereof as an active ingredient;
[Formula 1]

In Formula 1, R ₁ is a C _1-8 alkyl group, and R ₂ to R ₅ are each independently a hydroxy group.