KR20230009739A - Food Composition Comprising Marmelo Extract for Preventing or Alleviating Metabolic Disease - Google Patents

Abstract

The present invention relates to: a therapeutic and/or food composition which contains a Cydonia oblonga extract as an active component, and thus can be used for preventing, alleviating and/or treating related diseases such as weight and/or body fat loss, lowering blood sugar, lowering blood cholesterol, and the like; and a manufacturing method thereof.

Description

Food Composition Comprising Marmelo Extract for Preventing or Alleviating Metabolic Disease}

A composition for weight and/or body fat reduction, a composition for lowering blood sugar, a composition for lowering blood cholesterol, and/or for preventing, improving, and/or treating obesity and/or metabolic diseases, comprising a quince extract as an active ingredient It's about the composition.

The number of obese people is rapidly increasing due to excessive energy accumulation in the body due to frequent intake of instant foods and a meat-oriented diet, and reduction of energy consumption due to lack of exercise.

As such, excessively stored energy in the body appears in the form of various 　diseases as well as obesity, and 　metabolic 　diseases including diseases such as obesity, diabetes, hyperlipidemia, and metabolic syndrome are typical.

Chlorogenic acid is one of the most consumed phenolic compounds in the human diet, and various studies have been conducted on the physiological activity of chlorogenic acid. Various physiological activities have been identified.

Anti-obesity drugs and metabolic disease treatments developed so far cause side effects by acting on sympathetic nerves or hormones, but there is a problem that the effect is insignificant in the case of herbal ingredients with few side effects.

Accordingly, there is a need for the development of drugs with excellent effects and fewer side effects.

Korean Patent Publication No. 10-2015-0057119 (2015.05.28)

Under the above background, the present inventors have repeatedly studied the anti-obesity effect of chlorogenic acid. As a result, the content of chlorogenic acid in the quince extract was confirmed, and the quince extract containing chlorogenic acid inhibited weight gain, improved lipid metabolism, improved obesity-related hormone levels, reduced blood sugar, and reduced blood cholesterol, etc., thereby confirming the present invention. completed.

One example provides a composition for weight and/or body fat reduction comprising a quince extract as an active ingredient.

Another example provides a composition for lowering blood sugar containing a quince extract as an active ingredient.

Another example provides a blood cholesterol lowering composition comprising a quince extract as an active ingredient.

Another example provides a composition for preventing, improving, and/or treating obesity comprising a quince extract as an active ingredient.

Another example provides a composition for preventing, improving, and/or treating metabolic diseases comprising a quince extract as an active ingredient.

The composition may be a pharmaceutical composition or a food composition (eg, health functional food).

Another example provides a weight loss method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need of weight and/or body fat reduction.

Another example provides a blood glucose lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a blood cholesterol lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a method for preventing, improving, and/or treating obesity, comprising administering a quince extract to a subject in need of prevention, improvement, and/or treatment of obesity.

Another example provides a method for preventing, improving, and/or treating metabolic diseases, comprising administering a quince extract to a subject in need of such prevention, improvement, and/or treatment of metabolic diseases.

In the present specification, the quince extract administered with a high-fat diet effectively suppresses the increase in body weight and/or body fat and adipose tissue weight, blood sugar, blood cholesterol, blood insulin and leptin content induced by the high-fat diet, By confirming that the size increase is inhibited, anti-obesity, weight and / or body fat reduction, blood sugar reduction, blood cholesterol reduction, prevention, improvement, and / or treatment of metabolic diseases of the quince extract are provided.

Accordingly, one example provides a composition for weight and/or body fat reduction comprising a quince extract as an active ingredient.

Another example provides a composition for lowering blood sugar containing a quince extract as an active ingredient.

Another example provides a blood cholesterol lowering composition comprising a quince extract as an active ingredient.

Another example provides a composition for preventing, improving, and/or treating obesity comprising a quince extract as an active ingredient.

Another example provides a composition for preventing, improving, and/or treating metabolic diseases comprising a quince extract as an active ingredient.

The composition may be a pharmaceutical composition or a food composition (eg, health functional food).

Another example provides a method for reducing body weight and/or body fat comprising administering a pharmaceutically effective amount of a quince extract to a subject in need of weight loss.

Another example provides a blood glucose lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a blood cholesterol lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a method for preventing, improving, and/or treating obesity, comprising administering a quince extract to a subject in need of prevention, improvement, and/or treatment of obesity. Another example provides a method for preventing, improving, and/or treating metabolic diseases, comprising administering a quince extract to a subject in need of such prevention, improvement, and/or treatment of metabolic diseases.

Hereinafter, the present application will be described in more detail.

Quince Extract

In the present specification, a quince extract is provided as an active ingredient for reducing body weight and/or body fat, reducing blood sugar, reducing blood cholesterol, preventing, improving, and/or treating obesity and/or metabolic diseases.

Quince ( Cydonia oblonga ) is a fruit tree of the Rosaceae family, and is also called quince, Aiva, or European quince, and has a similar appearance to the fruit of quince and apple, but is distinguished from them.

Quince extract is all or part of quince (eg, at least one selected from the group consisting of fruits, seeds, pulp, peel, leaves, stems, roots, etc.) It may be a solvent extract extracted using.

The quince extract may be prepared by removing the quince juice after pressing the quince and extracting the remaining solid quince with an extraction solvent, but is not limited thereto.

In the present specification, "quince juice" means a liquid in the form of juice generated by pressing a quince fruit.

The extraction solvent may be at least one selected from the group consisting of water and straight-chain or branched-chain alcohol having 1 to 4 carbon atoms (eg, ethanol, fermented alcohol). In one embodiment, the extraction solvent may be at least one selected from the group consisting of water and an aqueous solution of straight-chain or branched-chain alcohol having 1 to 4 carbon atoms (e.g., aqueous ethanol solution, fermented alcohol), and the linear or branched-chain alcohol solution having 1 to 4 carbon atoms. Or branched chain alcohol aqueous solution, for example, 10 to 70 (v / v)%, 15 to 70 (v / v)%, 20 to 70 (v / v)%, 25 to 70 (v / v)%, 30 to 70 (v/v)%, 10 to 65 (v/v)%, 15 to 65 (v/v)%, 20 to 65 (v/v)%, 25 to 65 (v/v)%, 30 to 65 (v/v)%, 10 to 60 (v/v)%, 15 to 60 (v/v)%, 20 to 60 (v/v)%, 25 to 60 (v/v)%, 30 to 60 (v/v)%, 10 to 55 (v/v)%, 15 to 55 (v/v)%, 20 to 55 (v/v)%, 25 to 55 (v/v)%, 30 to 55 (v/v)%, 10 to 50 (v/v)%, 15 to 50 (v/v)%, 20 to 50 (v/v)%, 25 to 50 (v/v)% or 30 to 50 (v/v)% of a straight-chain or branched-chain alcohol having 1 to 4 carbon atoms (eg, ethanol, alcohol, fermented alcohol).

The amount of the extraction solvent used for the extraction is 1 to 10 times the volume of quince, 2 to 10 times, 3 to 10 times, 4 to 10 times, 1 to 9 times, 2 to 9 times, 3 to 9 times the volume, 4 to 9 times the volume, 1 to 8 times the volume, 2 to 8 times the volume, 3 to 8 times the volume, 4 to 8 times the volume, 1 to 7 times the volume, 2 to 7 times the volume, 3 to 7 times the volume It may be 4 to 7 times by volume, 1 to 6 times by volume, 2 to 6 times by volume, 3 to 6 times by volume, or 4 to 6 times by volume, but is not limited thereto.

The extraction time is, but is not limited to, 1 to 24 hours, 1 to 12 hours, 1 to 6 hours, 1 to 5 hours, 3 to 24 hours, 3 to 12 hours, 3 to 6 hours for sufficient extraction of active ingredients. time, 3 to 5 hours, 5 to 24 hours, 5 to 12 hours or 5 to 6 hours. The extraction temperature is, but is not limited to, a temperature range selected from room temperature to the boiling point of the extraction solvent, for example, 50 to 100 ° C, 55 to 100 ° C, 60 to 100 ° C, 65 to 100 ° C, for effective extraction of active ingredients. , 50 to 95 ℃, 55 to 95 ℃, 60 to 95 ℃, 65 to 95 ℃, 50 to 90 ℃, 55 to 90 ℃, 60 to 90 ℃, 65 to 90 ℃, 50 to 85 ℃, 55 to 85 ℃ , 60 to 85 °C, 65 to 85 °C, 50 to 80 °C, 55 to 80 °C, 60 to 80 °C or 65 to 80 °C.

An extraction method for obtaining the extract may be selected from all known extraction methods commonly used in the art to which the present invention belongs. For example, conventionally used extraction methods such as reflux extraction, immersion extraction, heating extraction, cold precipitation, hot water extraction, ultrasonic extraction, filtration, pressurized extraction, supercritical extraction, and electrical extraction may be used.

The extraction may be performed once, or may be performed two or more times (eg, 2, 3, 4, or 5 times) with the same or different extraction solvents and/or conditions. When the extraction is performed two or more times, the extraction conditions such as the extraction solvent used in each extraction, the extraction time for each extraction, and the temperature may be adjusted identically or differently within the range described above, but are not limited thereto.

In one embodiment, the extract is 10 to 100 brix, 20 to 100 brix, 30 to 100 brix, 40 to 100 brix, 50 to 100 brix, 60 to 100 brix, 70 to 100 brix, 80 to 100 brix, 90 to 100 brix 100 brix, 10 to 90 brix, 20 to 90 brix, 30 to 90 brix, 40 to 90 brix, 50 to 90 brix, 60 to 90 brix, 70 to 90 brix, 80 to 90 brix, 10 to 80 brix, 20 to 80 brix, 30 to 80 brix, 40 to 80 brix, 50 to 80 brix, 60 to 80 brix, 70 to 80 brix, 10 to 70 brix, 20 to 70 brix, 30 to 70 brix, 40 to 70 brix, 50 to It may have a sugar content of 70 brix or 60 to 70 brix, but is not limited thereto.

In the present specification, the quince extract is not only the solvent extract described above, but also the solvent extract is concentrated by a conventional method (eg, vacuum concentration, vacuum concentration, etc.), dried (eg, spray drying, freeze-drying, etc.), and / or pulverized (powdered) concentrates, dried substances, and/or pulverized substances.

The quince extract provided as an active ingredient in the present specification has a chlorogenic acid content measured by HPLC of 0.2 mg/g or more, 0.3 mg/g or more, based on the total weight of the extract (eg, concentrate weight). 0.4 mg/g or more, 0.5 mg/g or more, 0.6 mg/g or more or 0.7 mg/g or more, such as 0.2 to 2 mg/g, 0.3 to 2 mg/g, 0.4 to 2 mg/g, 0.5 to 2 mg/g, 0.6 to 2 mg/g, 0.7 to 2 mg/g, 0.8 to 2 mg/g, 0.2 to 1.5 mg/g, 0.3 to 1.5 mg/g, 0.4 to 1.5 mg/g, 0.5 to 1.5 mg/g, 0.6 to 1.5 mg/g, 0.7 to 1.5 mg/g, 0.2 to 1.2 mg/g, 0.3 to 1.2 mg/g, 0.4 to 1.2 mg/g, 0.5 to 1.2 mg/g, 0.6 to 1.2 mg/g, 0.7 to 1.2 mg/g, 0.2 to 1.0 mg/g, 0.3 to 1.0 mg/g, 0.4 to 1.0 mg/g, 0.5 to 1.0 mg/g, 0.6 to 1.0 mg/g, 0.7 to 1.0 mg /g, 0.2 to 0.8 mg/g, 0.3 to 0.8 mg/g, 0.4 to 0.8 mg/g, 0.5 to 0.8 mg/g, 0.6 to 0.8 mg/g or 0.7 to 0.8 mg/g.

medicinal use

One embodiment provides a pharmaceutical composition for reducing body weight and / or body fat containing a quince extract as an active ingredient.

Another example provides a pharmaceutical composition for lowering blood sugar containing a quince extract as an active ingredient.

Another example provides a pharmaceutical composition for lowering blood cholesterol containing a quince extract as an active ingredient.

Another example provides a pharmaceutical composition for preventing, improving, and/or treating obesity comprising a quince extract as an active ingredient.

Another example provides a pharmaceutical composition for preventing, improving, and/or treating metabolic diseases comprising a quince extract as an active ingredient.

Another example provides a weight loss method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need of weight and/or body fat reduction.

Another example provides a blood glucose lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a blood cholesterol lowering method comprising administering a pharmaceutically effective amount of a quince extract to a subject in need thereof.

Another example provides a method for preventing, improving, and/or treating obesity, comprising administering a pharmaceutically effective amount of a quince extract to a subject in need of prevention, improvement, and/or treatment of obesity.

Another embodiment provides a method for preventing, improving, and/or treating metabolic diseases, comprising administering a pharmaceutically effective amount of a quince extract to a subject in need of such prevention, improvement, and/or treatment of metabolic diseases. .

A pharmaceutically effective amount of the quince extract can be used to (1) reduce body weight and/or body fat, (2) lower blood sugar, (3) lower blood cholesterol, and/or (4) prevent, improve, and/or prevent obesity and/or metabolic diseases. Alternatively, in the step of administering to a subject in need of treatment, the content of the quince extract is 1 to 400 mg/kg (body weight; BW), 50 to 400 mg/kg, 100 to 400 mg based on the body weight of the subject to be administered. /kg, 150 to 400 mg/kg, 200 to 400 mg/kg, 1 to 300 mg/kg (body weight; BW), 50 to 300 mg/kg, 100 to 300 mg/kg, 150 to 300 mg/kg , 200 to 300 mg/kg, 1 to 200 mg/kg (body weight; BW), 50 to 200 mg/kg, 100 to 200 mg/kg, and 150 to 200 mg/kg may be administered, limited thereto. it is not going to be

A pharmaceutically effective amount of the quince extract can be used to (1) reduce body weight and/or body fat, (2) lower blood sugar, (3) lower blood cholesterol, and/or (4) prevent, improve, and/or prevent obesity and/or metabolic diseases. Alternatively, in the step of administering to a subject in need of treatment, if the subject is a human, the dose may be determined by calculating by applying a body conversion factor (0.08). Accordingly, in the step of administering to a human in need of prevention, improvement, and/or treatment of obesity and/or metabolic disease, the content of the quince extract is 0.0048 to 1.92 g based on 60 kg of the body weight of the human to be administered. /day, 0.24 to 1.92 g/day, 0.48 to 1.92 g/day, 0.72 to 1.92 g/day, 0.96 to 1.92 g/day, 0.0048 to 1.44 g/day, 0.24 to 1.44 g/day, 0.48 to 1.44 g/day 0.72 to 1.44 g/day, 0.96 to 1.44 g/day, 0.0048 to 0.96 g/day, 0.24 to 0.96 g/day, 0.48 to 0.96 g/day or 0.72 to 0.96 g/day, It is not limited thereto.

The quince extract was as described above.

The blood glucose lowering refers to a concept including a decrease in blood glucose concentration, a decrease in blood insulin content, a decrease in insulin resistance, and/or an increase in insulin sensitivity. Insulin resistance can be calculated by Equation 2 of Example 6.3 below, and insulin sensitivity can be calculated by Equation 3 of Example 6.3 below.

The lowering of blood cholesterol means a decrease in total cholesterol content or low-density lipoprotein (LDL) cholesterol content in blood.

The obesity refers to a state in which body fat is abnormally large, and refers to a state in which a large amount of calories are consumed but not consumed and stored in the form of fat in the body.

In one example, obesity may mean a state in which a body mass index (BMI; weight divided by height squared; kg/m ² ) is about 25 kg/m ² or more, but is not limited thereto.

The metabolic disease may be a general term for diseases that cause abnormalities in each organ as metabolites accumulate in the body or specific organs in the body due to disorders or functional abnormalities of hormones or internal organs involved in metabolism.

In one example, the metabolic disease may be a disease caused by the accumulation of one or more types selected from the group consisting of glucose, cholesterol, and triglycerides in blood or all of them. For example, the metabolic disease may be diabetes, hyperlipidemia, atherosclerosis, hyperinsulinemia or metabolic syndrome.

Diabetes mellitus is a kind of systemic metabolic disease caused by genetic or environmental factors. It is a disease caused by an absolute or relative lack of insulin in the body, and refers to a state in which blood sugar concentration is abnormally high. In the present specification, diabetes may include all of type 1 diabetes, type 2 diabetes, and gestational diabetes, and in particular, may be type 2 diabetes related to insulin resistance.

Hyperlipidemia may refer to a condition in which more than necessary fat substances are present in the blood or cause inflammation. For example, hyperlipidemia can be diagnosed when fasting serum cholesterol is 220 　mg/dl or more or triglyceride is 150 　mg/dl or more, but is not limited thereto. Hyperlipidemia may refer to hypercholesterolemia, hypertriglyceridemia, or both.

Arteriosclerosis is a general term for diseases in which arterial blood vessels are narrowed for reasons such as the formation of blood clots in arteries. In the present specification, it may be related to high cholesterol and/or triglyceride content in blood.

Hyperinsulinemia refers to a state in which blood insulin levels are high, and is characterized by organic hyperinsulinemia caused by proliferation (adenoma, hypertrophy) of islets of Langerhans, and functional disorders caused by dysfunction of the autonomic nervous system and digestive system without islet adenoma. It can be divided into hyperinsulinemia.

Metabolic syndrome refers to a syndrome in which risk factors such as hyperlipidemia, hypertension, abnormal glucose metabolism, and obesity appear together. Metabolic syndrome is a disease presumed to be caused by insulin resistance.

As used herein, 'treatment' includes improvement, alleviation or stabilization of a disease state or symptom, partial or complete recovery, prolongation of survival, reduction of disease extent, delay or alleviation of disease progression, and other beneficial treatment results. used in meaning 'Prevention' is used in the meaning of including all mechanisms and/or effects that prevent the occurrence of a specific disease by acting on a subject who does not have a specific disease, delay the onset of the specific disease, or reduce the incidence of the specific disease.

The content of the quince extract used as an active ingredient in the pharmaceutical composition provided herein can be appropriately adjusted depending on the type and purpose of use, the condition of the subject to be used, the type and severity of symptoms, etc., and the solid content (after removing the solvent from the extract) Solid component) 0.001 to 99.9% by weight, 0.001 to 90% by weight, 0.001 to 75% by weight, 0.001 to 50% by weight, 0.01 to 99.9% by weight, 0.01 to 90% by weight, 0.01 to 75% by weight, 0.01 to 0.01% by weight 50 wt%, 0.1 to 99.9 wt%, 0.1 to 90 wt%, 0.1 to 75 wt%, 0.1 to 50 wt%, 1 to 99.9 wt%, 1 to 90 wt%, 1 to 75 wt%, 1 to 50 wt% %, 5 to 99.9%, 5 to 90%, 5 to 75%, 5 to 50%, 10 to 99.9%, 10 to 90%, 10 to 75%, or 10 to 50% It may be, but is not limited thereto.

The pharmaceutical composition or active ingredient, the quince extract, is used in humans, primates such as monkeys, rodents such as mice, rats, rabbits, other mammals including dogs, cats, cows, pigs, sheep, horses, goats, chickens, etc. , birds including ducks and geese, reptiles including snakes, lizards, turtles, crocodiles, etc., amphibians, and vertebrates such as fish, etc., may be administered to an administration subject selected from various routes. The method of administration of the pharmaceutical composition may be any method commonly used, for example, oral administration, or parenteral administration such as intravenous, intramuscular, subcutaneous, or intraperitoneal injection may be administered. In one example, the pharmaceutical composition may be for oral administration, but is not limited thereto.

The pharmaceutical composition may be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, or parenteral formulations in the form of sterile injectable solutions according to conventional methods, respectively. there is.

In addition to the active ingredients described above, the pharmaceutical composition is generally selected from the group consisting of pharmaceutically and / or physiologically acceptable carriers, excipients, diluents, etc., selected in consideration of the method of administration and standard pharmaceutical practice. It may be administered in combination with one or more selected adjuvants. For example, the pharmaceutically and/or physiologically acceptable carrier may be lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, It may contain at least one selected from the group consisting of cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. However, it is not limited thereto, and may be any carrier commonly used in the pharmaceutical field.

The pharmaceutically and / or physiologically acceptable diluent and / or excipient is from the group consisting of all fillers, extenders, binders, wetting agents, disintegrants, lubricants, surfactants, etc. commonly used in the appropriate formulation of pharmaceutical compositions. It may be one or more selected species.

For example, in the case of solid preparations for oral administration such as tablets, pills, powders, granules, or capsules, the excipient is at least one material for formulating such solid preparations, such as starch, calcium carbonate , sucrose, lactose, and may be one or more selected from the group consisting of gelatin. In addition, lubricants such as magnesium styrate, talc, and the like may also be used. In addition, in the case of formulation of liquid preparations for oral administration such as suspensions, internal solutions, emulsions, syrups, etc., at least one selected from the group consisting of commonly used simple diluents such as water, liquid paraffin, etc. may be used. , Various excipients commonly used, for example, wetting agents, sweeteners, aromatics, and / or preservatives may be included, but are not limited thereto.

For example, the pharmaceutical composition may be in the form of a tablet containing starch or lactose, or in the form of a capsule containing suitable excipients, or in the form of an elixir or suspension containing a flavoring or coloring chemical, orally, orally. It can be administered intra- or sublingually. Such liquid formulations may be prepared by suspending agents (e.g. methylcellulose, semi-synthetic glycerides such as Witepsol, or mixtures of Apricot kernel oil and PEG-6 esters or PEG-8 and caprylic/capric acid). It can be formulated with pharmaceutically acceptable excipients such as mixtures of glycerides), but is not limited thereto.

food composition

One example provides a food composition for weight and / or body fat reduction containing a quince extract as an active ingredient.

Another example provides a food composition for lowering blood sugar containing a quince extract as an active ingredient.

Another example provides a food composition for lowering blood cholesterol containing a quince extract as an active ingredient.

Another example provides a food composition for preventing and/or improving obesity comprising a quince extract as an active ingredient.

Another example provides a food composition for preventing and/or improving metabolic diseases comprising a quince extract as an active ingredient.

Another example is a food composition comprising a quince extract as an active ingredient for (1) weight and/or body fat reduction, (2) blood sugar lowering, (3) blood cholesterol lowering and/or (4) obesity and/or metabolic disease. Provided is a method for weight loss comprising administering to a subject in need of prevention, improvement and/or treatment.

The food composition may be used for (1) reducing body weight and/or body fat, (2) lowering blood sugar, (3) lowering blood cholesterol, and/or (4) preventing, improving, and/or treating obesity and/or metabolic disease. In the step of administering to the subject, the content of the quince extract is 1 to 400 mg/kg (body weight; BW), 50 to 400 mg/kg, 100 to 400 mg/kg, 150 to 400 based on the body weight of the subject to be administered. mg/kg, 200 to 400 mg/kg, 1 to 300 mg/kg (body weight; BW), 50 to 300 mg/kg, 100 to 300 mg/kg, 150 to 300 mg/kg, 200 to 300 mg/kg kg, 1 to 200 mg/kg (body weight; BW), 50 to 200 mg/kg, 100 to 200 mg/kg, and 150 to 200 mg/kg may be administered, but is not limited thereto.

The food composition may be used for (1) reducing body weight and/or body fat, (2) lowering blood sugar, (3) lowering blood cholesterol, and/or (4) preventing, improving, and/or treating obesity and/or metabolic disease. In the step of administering to a subject, if the subject is a human, the dosage may be determined by calculating by applying a body conversion factor (0.08). Accordingly, in the step of administering to a human in need of prevention, improvement, and/or treatment of obesity and/or metabolic disease, the content of the quince extract is 0.0048 to 1.92 g based on 60 kg of the body weight of the human to be administered. /day, 0.24 to 1.92 g/day, 0.48 to 1.92 g/day, 0.72 to 1.92 g/day, 0.96 to 1.92 g/day, 0.0048 to 1.44 g/day, 0.24 to 1.44 g/day, 0.48 to 1.44 g/day 0.72 to 1.44 g/day, 0.96 to 1.44 g/day, 0.0048 to 0.96 g/day, 0.24 to 0.96 g/day, 0.48 to 0.96 g/day or 0.72 to 0.96 g/day, It is not limited thereto.

The quince extract, lowering blood sugar, lowering blood cholesterol, and metabolic diseases are as described above.

In the food composition provided herein, the term "food" means an edible natural product or processed product containing one or more nutrients, and as a conventional meaning, various general foods, health functional foods, beverages, food additives, beverages It may be used to mean one or more selected from the group consisting of additives and the like. The term “food composition” may refer to a combination of ingredients for preparing the food product. In one example, the food may mean health functional food.

The content of the quince extract in the food composition can be appropriately adjusted depending on the type of food use, purpose, etc., for example, 0.00001 to 99.9% by weight, 0.0001 to 99.9% by weight, 0.001 to 99.9% by weight, or 0.001% by weight based on the solid content. to 50% by weight, but is not limited thereto.

The administration 'subject' of the food composition is a human, primates such as monkeys, rodents such as mice, rats, rabbits, other mammals including dogs, cats, cows, pigs, sheep, horses, goats, chickens, ducks, Birds including geese, reptiles including snakes, lizards, turtles and crocodiles, amphibians, and vertebrates such as fish, cells, tissues, or cell cultures isolated from the objects.

The quince extract provided herein reduces or effectively suppresses the increase in body weight and / or body fat and adipose tissue weight, blood sugar, blood cholesterol, blood insulin and leptin, thereby reducing obesity and improving lipids. It further suggests that there is a preventive, therapeutic, and / or ameliorative effect on metabolic diseases, and future anti-obesity efficacy and / or a preventive, therapeutic, and / or ameliorative effect on metabolic diseases Possibility of development as a health functional material presents

1a and 1b are graphs showing the results of analyzing the chlorogenic acid content in the quince extract by HPLC chromatography. Figure 1a shows the chlorogenic acid standard form, Figure 1b shows the results of sample c (Table 4 in Example 2.2).
Figure 2 is a graph showing the weekly weight change according to the type of diet. Group 1 is the control diet group, group 2 is the high-fat diet group, and group 3 is the high-fat diet group administered with mdB-44.
3 is a photograph of histological changes in epididymal adipose tissue observed using an optical microscope (Carl Zeiss).

Hereinafter, the present invention will be described in more 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 quince extract

Quince fruits (LICORICE EXTRACT (Tashkent, Republic of Uzbekistan)) were weighed, sorted, washed and decontaminated. Thereafter, the quince was pressed with a press at a pressure of 1 to 5 kg/cm ² to remove quince juice (liquid in the form of juice produced by pressing the quince fruit), and the solids were separated. The solid was extracted twice for 5 to 6 hours at 65 to 80 ° C. with 30 to 50 (v / v)% alcohol in an amount of 4 to 6 volumes. The extract was filtered through a 10 μm standard filter and vacuum concentrated to 65 brix or more using a vacuum evaporator (EYELA, Tokyo, Japan) to obtain a quince concentrate. The concentrate was powdered through a spray dryer. The extract or powdered quince extract prepared as described above was used in the following examples.

Example 2. Analysis of chlorogenic acid content of quince extract

2.1 HPLC analysis method

The chlorogenic acid (CGA) content of the quince extract prepared in Example 1 was analyzed using an LC-20A high-performance liquid chromatography (HPLC) system (Shimadzu, Japan). HPLC analysis conditions are shown in Table 1 (analysis conditions) and Table 2 (mobile phase gradient conditions) below.

Items Conditions Column Capcell Pak C ₁₈ column (Shiseido, 5μm, 250×4.6mm) Column Temperature 40℃ Flow 1.4 mL/min Detector Diode Array Detector(DAD) 330 nm Injection Volume 5 μL Mobile Phase A: phosphoric acid : water = 0.5 : 99.5 (v/v),
B : phosphoric acid : AcN = 0.5 : 99.5 (v/v) Run time 40min

Time (Min) A solvent (%) B solvent (%) 0 95 5 7 95 5 27 70 30 28 10 90 30 10 90 31 95 5 40 95 5

5 mg of chlorogenic acid standard was weighed and dissolved in 50 mL of methanol. 5mL of this solution was taken and transferred to a volumetric flask, and 5mL of methanol was added, and 20mL of distilled water was added thereto and used as a standard stock solution. The solution of the stock solution was diluted step by step using the standard stock solution to prepare a calibration curve to prepare a chlorogenic acid standard solution (final concentration: 0.00833, 0.00416, 0.00208, 0.00104, 0.01666 mg / mL), and the corresponding information is shown in Table 3 below. In addition, 30 mg of the powdered quince extract of Example 1 was weighed, dissolved in 5mL of 30% methanol, and filtered through a 0.45um filter to be used as a test solution (final concentration 30mg/5mL) (quince extract). The corresponding information is shown in Table 3 below.

sample Pretreatment concentration (Concentration) Chlorogenic acid standard solution 0.00833, 0.00416, 0.00208, 0.00104, 0.01666 mg/mL in 30% MeOH Quince Extract 30mg/5mL in 30% MeOH (HPLC grade)

2.2 Experimental results

The experiment was performed three times by the HPLC analysis method of 2.1 above, and the obtained values are shown in Table 4, Fig. 1a (chlorogenic acid standard form) and 1b (sample c).

sample name Content (mg/g) sample a 0.82 sample b 0.7 sample c 0.73

(Samples a, b, and c refer to test solutions containing the quince extract used in each experiment when the above experiment is performed three times)

As shown in Table 4 and FIGS. 1A and 1B, the chlorogenic acid content in the quince extract prepared in Example 1 was 0.82 mg/g for sample a, 0.7 mg/g for sample b, and 0.73 mg for sample c. /g, it was confirmed that the distribution was in the range of ±20% based on the case of sample c, which had a median value of chlorogenic acid content. The results show that the quince extract obtained in Example 1 contains chlorogenic acid in a certain range (e.g., 0.75 (mg/g) ± 20%), and this result indicates that even after performing the extraction several times, the chlorogenic acid contained in the extract It shows that the acid content is in the same range.

Example 3. Preparation of test substances and experimental animals

3.1 Test substance preparation and animal test approval

As a test substance, the powdered quince extract (powder; hereinafter, 'mdB-44') prepared in Example 1 was used, and all animal experiments in the following examples were conducted under the approval of the Animal Experimentation Ethics Committee of Hallym University. It was performed according to regulations (Hallym 2019-67).

3.2 Test group and test substance administration

Five-week-old, male C57BL/6 mice free from specific pathogens were purchased from Dooyeol Biotech Co., Ltd. and used. After a week of quarantine and adaptation, healthy animals without weight loss were selected and used in the experiment. Experimental animals were raised in a breeding environment set at a temperature of 23 ± 3 ° C, relative humidity of 50 ± 10%, ventilation frequency of 10 to 15 times / hour, lighting time of 12 hours (08:00 to 20:00), and illumination of 150 to 300 Lux. . During the adaptation period, the experimental animals were allowed to freely consume solid feed for experimental animals (Cargill Agripurina Co., Ltd.) and drinking water. After a 1-week adaptation period, healthy animals were selected and subjected to control diet (CD) (Group 1), high-fat diet (HFD) control group (Group 2), high-fat diet + They were classified into the 200mg/kg body weight (BW) mdB-44 (test substance of Example 3.1) administration group (Group 3). During the entire test period, the energy ratio (kcal%) (protein:carbohydrate:fat) of the control diet was 20:70:10, and the energy ratio (kcal%) (protein:carbohydrate:fat) of the high-fat diet was 20:20: 60, and both diets are from Research Diets, Inc. (New Brunswick, NJ, USA). The composition of the control diet group and the high-fat diet are shown in Table 5, and the experimental design is shown in Table 6.

Control Diet (CD) (10 kcal% fat) Content of Each Composition (g) Control diet (CD) (10 kcal% fat) Calorie content of each composition (kcal) High-fat diet (HFD) (60 kcal% fat) Content of each composition (g) High Fat Diet (HFD) (60 kcal% fat) Calorie content of each composition (kcal) Casein, 80 Mesh 200 800 200 800 L-Cystine 3 12 3 12 Corn starch 315 1260 0 0 Maltodextrin 10 35 140 125 500 Sucrose 350 1400 68.8 275.2 Cellulose, BW200 50 0 50 0 Soybean oil 25 225 25 225 Lard* 20 180 245 2205 Mineral mix 10 0 10 0 Dicalcium phosphate 13 0 13 0 Calcium carbonate 5.5 0 5.5 0 Potassium citrate, 1H ₂ O 16.5 0 16.5 0 Vitamin mix V10001 10 40 10 40 Choline bitartrate 2 0 2 0 Sum 1055.05 4057 773.85 4057

(*Lard: Cholesterol is calculated as 0.95mg/g)

group 1 group 2 group 3 control group (normal) negative control group experimental group empirical diet CD HFD HFD test substance
(mg/kg BW) - - mdB-44 200 number of animals 10 10 10

Drinking water was freely consumed during the entire test period, and the test substance of Example 3.1 was dissolved in drinking water at 200 mg per 1 kg of experimental animals and orally administered at a certain time for 8 weeks (Group 3). Statistical processing in the following examples using the corresponding experimental animals was expressed as mean ± standard error. The collected results were analyzed using the GraphPad Prism 5.0 (GraphPad software) program. Student's t-test and one-way analysis of variance (ANOVA) were used to compare the difference between the test substance administration group and the control group. Only when p < 0.05 or more was judged statistically significant.

Example 4. Measurement of body weight and food intake

The body weights of the experimental animals prepared in Example 3 were measured at 0 and 8 weeks, and food intake was measured every 2 days for 8 weeks. The daily food intake and daily energy intake were calculated by calculating the intake amount during the entire test period. Food efficiency ratio (FER) was calculated by dividing the weight gain during the test period by the amount of food consumed during the same period as shown in Equation 1 below.

[Equation 1]

Dietary efficiency = weight gain (g) / dietary intake (g)

The measured weight results and the resulting total weight gain and daily weight gain are shown in Table 7, and the dietary intake and dietary efficiency measured for 8 weeks are shown in Table 8, and in particular, the weight results are shown in FIG. 2 .

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 0 weeks 23.14 ± 0.35 23.06 ± 0.24 23.10 ± 0.27 8 weeks 31.20 ± 0.67 41.91 ± 0.91 ^*** 36.45 ± 0.91 ^### Total weight gain (g) 8.1 ± 0.7 18.9±0.9 ^*** 13.4±1.1 ^## Daily weight gain (g/day) 0.146 ± 0.013 0.343 ± 0.016 ^*** 0.243 ± 0.020 ^##

(The results in Table 8 are expressed as mean ± standard error, ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 Group 1 (normal control group); Group 2 (negative control group); Group 3: experimental group, (mdB-44 administration group)

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 Total dietary intake (g) 145.4 ± 1.4 129.1 ± 1.2 ^*** 112.7 ± 1.5 ^### Daily dietary intake (g/day) 2.64 ± 0.03 2.35±0.02 ^*** 2.05 ± 0.03 ^### Daily energy intake (kcal/day) 10.18 ± 0.10 12.30 ± 0.11 ^*** 10.73 ± 0.15 ^### Dietary efficiency (weight gain/diet intake) 0.055 ± 0.005 0.146 ± 0.007 ^*** 0.119 ± 0.009 ^#

(The above figures are expressed as mean ± standard error, and mean ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 )

As shown in Tables 7 and 8, during the test period, the experimental animals of all test groups continuously increased their body weight and showed a normal body weight change. Compared to the control group (Group 1), the weight of the high-fat diet control group (Group 2) increased significantly more. On the other hand, in group 3, the test group administered with the quince extract, the body weight (weight gain) of the experimental animals increased by the high-fat diet was significantly decreased compared to the control group (group 2) fed the high-fat diet. Through this, it was confirmed that even if a high-fat diet was consumed in parallel with the quince extract, weight gain was suppressed.

In addition, the food intake of group 3, the test group administered with the quince extract, decreased significantly compared to the negative control group (group 2), but the dietary efficiency (weight gain/food intake) also decreased, indicating that only a decrease in food intake reduced body weight. does not mean that it has decreased.

Example 5. Measurement of body fat

After anesthetizing the experimental animals 1 day before the end of the experimental animal test in Example 3.2, body composition components were measured using dual-energy X-ray absorptiometry (DEXA, PIXImusTM, GE Lunar) to evaluate body fat percentage, and the results are in the table 9.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 Fat (%) 29.58 ± 0.84 42.84 ± 1.66 ^*** 37.97 ± 1.46 ^#

(The above figures are expressed as mean ± standard error, and mean ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 )

The body fat percentage in the high-fat diet control group (Group 2) was 42.84 ± 1.66% compared to 29.58 ± 0.84% in the control group (Group 1), and the body fat percentage significantly increased compared to the high-fat diet control group (Group 2), mdB- 44 The percentage of body fat decreased significantly in the 200mg/kg BW group (Group 3).

Example 6. Blood and fat analysis

Example 6.1 Blood collection and tissue extraction

After completion of the test of the test animal in Example 3, the test animal was sacrificed. Before sacrifice, orbital blood was collected after anesthesia using an anesthetic prepared by diluting tribromoethanol with tert-amyl alcohol. The blood was placed in a serum separate tube (Becton Dickinson), left at room temperature for 30 minutes, and then centrifuged at 5,000 rpm for 10 minutes to separate the serum and stored at -70°C until analysis. After blood collection, the experimental animals were sacrificed, and white adipose tissue (epididymal fat, visceral fat, retro-abdominal fat, inguinal fat) was extracted, rinsed with cold physiological saline, and after removing excess water with filter paper, the weight was measured. Epididymal adipose tissue was divided into three parts, some were fixed in 4% paraformaldehyde (PFA), embedded in paraffin, and tissue staining was performed. Some of them were subjected to real-time RT-PCR after total RNA was isolated. After separation, Western blot was performed and stored at -70°C for future further analysis.

6.2 Tissue (adipose) weighing

The weight of the adipose tissue was measured in Example 6.1, and the results are shown in Table 10.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 epididymal fat 1.030 ± 0.070 2.471 ± 0.079 ^*** 2.023 ± 0.109 ^## retroperitoneal fat 0.447 ± 0.037 1.184 ± 0.063 ^*** 0.954 ± 0.074 ^# visceral fat 0.603 ± 0.050 1.603 ± 0.114 ^*** 0.975 ± 0.073 ^### Seohyebu region 0.210 ± 0.031 0.544 ± 0.033 ^*** 0.490 ± 0.052 total weight of white fat 2.29 ± 0.14 5.80±0.21 ^*** 4.44 ± 0.21 ^###

(The above figures are expressed as mean ± standard error, and mean ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 )

Body fat is divided into white adipose tissue and brown adipose tissue according to its shape or function. White adipose tissue mainly stores surplus energy in the body as fat, and brown adipose tissue produces heat, and the weight of white adipose increases. The more you do it, the more weight you gain. The weights of epididymal fat, retroperitoneal fat, visceral fat, and inguinal fat, which are white adipose tissues, were all significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1). On the other hand, group 3 administered with the quince extract showed a decrease in body fat in all items tested compared to the high-fat diet control group (group 2).

6.3 Analysis of blood glucose related values

The glucose content in the blood collected in Example 6.1 was measured using a blood biochemical analyzer (KoneLab 20 XT, Thermo Fisher Scientific), and the insulin content was presented by the manufacturer using a measurement kit purchased from Millipore Corporation. It was measured according to one method.

Since insulin resistance is the most important factor in metabolic syndrome, measurement of blood insulin concentration is very useful for measuring insulin sensitivity and resistance, and in this study, insulin resistance (HOMA-IR) and insulin The sensitivity (QUICKI) was calculated. Insulin resistance (homeostatic model assessment for insulin resistance, HOMA-IR) and insulin sensitivity (quantitative insulin sensitivity check index, QUICKI) were calculated using Equations 2 and 3 below using fasting blood glucose and blood insulin content, and the measurement The results are shown in Table 11.

[Equation 2]

Insulin resistance (HOMA-IR) = [insulin (mU/L) × glucose (mg/dL)] / 405

[Equation 3]

Insulin sensitivity (QUICKI) = 1 / [log insulin (mU/L) + log glucose (mg/dL)]

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 Glucose (mg/dL) 149.0 ± 10.8 196.8 ± 7.8 ^** 185.2 ± 7.5 Insulin (ng/mL) 2.62 ± 0.21 9.12 ± 0.97 ^*** 5.67±0.81 ^# HOMA-IR 22.9 ± 2.2 107.1 ± 12.4 ^*** 62.3±9.3 ^## QUICKI 0.253 ± 0.002 0.218 ± 0.004 ^*** 0.230 ± 0.003 ^#

Glucose was significantly increased in the high fat diet control group (group 2) compared to the control group (group 1), and decreased in group 3 compared to the high fat diet control group (group 2).

Blood insulin content increased significantly to 9.12 ± 0.97ng/mL in the high-fat diet control group (Group 2) compared to 2.62 ± 0.21ng/mL in the control group (Group 1), whereas in the mdB-44 administration group (Group 3) 5.67 ± 0.81 ng / mL, the high-fat diet significantly decreased compared to the control group.

Insulin resistance significantly increased from 22.9 ± 2.2 in the control group (Group 1) to 107.1 ± 12.4 in the high-fat diet control group (Group 2), whereas in the mdB-44 administration group (Group 3), the high-fat diet decreased compared to the control group. . Insulin sensitivity was significantly reduced to 0.218 ± 0.004 in the high-fat diet control group (Group 2) compared to 0.253 ± 0.002 in the control group (Group 1), whereas in the mdB-44 administration group (Group 3), the high-fat diet compared to the control group increased. Through this, it was confirmed that in the case of the quince extract-administered group, impaired glucose tolerance was improved by suppressing insulin resistance caused by a high-fat diet.

6.4 Numerical analysis of fat

The contents of triglyceride (TG), total cholesterol (CHOL), and HDL-cholesterol (HDL-CHOL) in the blood collected in Example 6.1 were measured using a blood biochemistry analyzer (KoneLab 20 XT, Thermo Fisher Scientific) was measured, and the arteriosclerosis index was calculated through Equation 4 below using the blood cholesterol content and the HDL-cholesterol content.

[Equation 4]

Atherosclerosis index = total cholesterol content / HDL-cholesterol content

The obtained results are shown in Table 12.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 TG (mg/dL) 77.1 ± 3.3 90.4±3.4 ^* 77.3±3.2 ^# CHOL (mg/dL) 165.7 ± 8.4 188.4 ± 4.7 ^* 171.8±5.3 ^# HDL-CHOL (mg/dL) 151.6 ± 6.2 126.9±5.8 ^** 150.9±6.1 ^# arteriosclerosis index 1.09 ± 0.04 1.52±0.11 ^** 1.15±0.04 ^##

In general, when a high-fat diet is ingested, the supply of fatty acids increases and the concentrations of triglyceride and total cholesterol in the blood increase, whereas the blood concentration of HDL-cholesterol tends to decrease, which causes cardiovascular and coronary vascular diseases.

Blood triglyceride and blood total cholesterol contents were significantly increased in the high-fat control group (Group 2) compared to the control group (Group 1), whereas in Group 3 administered with mdB-44, there was a significant increase compared to Group 2. decreased, and blood triglyceride levels were measured very similarly to those of the control group (Group 1). Blood HDL-cholesterol content was significantly decreased in the high-fat diet control group (group 2) compared to the control diet group (group 1), whereas it was significantly increased in group 3 administered with mdB-44 compared to group 2, It was measured very similarly to the control group (group 1). As a result of calculating the arteriosclerosis index (see Equation 4), which is the most effective predictor of coronary artery disease, using the blood cholesterol content and the HDL-cholesterol content, it was compared with 1.09±0.04 of the control diet group (Group 1). The control group (Group 2) significantly increased to 1.52 ± 0.11, while the mdB-44 200mg/kg BW group (Group 3) significantly decreased to 1.15 ± 0.04 (Group 3) compared to Group 2.

6.5 Analysis of serum leptin and adiponection content

Adipose tissue is recognized as an endocrine organ that affects the metabolic process of the body by synthesizing and secreting various adipokines that play a very important physiological role as well as a place to store surplus energy. Leptin, one of the representative adipokines secreted from adipose tissue, is an important substance regulating energy intake and storage, insulin sensitivity, and metabolic rate. Another major adipokine, adiponectin, is known as an important hormone that increases fatty acid oxidation in muscle and prevents chronic diseases such as diabetes and metabolic syndrome through its role as an anti-inflammatory cytokine.

The content of leptin and adiponectin in serum of the blood collected in Example 6.1 was measured using a measurement kit purchased from R&D Systems according to the method suggested by the manufacturer, and the results are shown in Table 13 below.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 Adiponectin (ng/mL) 10.45 ± 0.26 9.45 ± 0.25 ^* 10.84±0.37 ^## Leptin (ng/mL) 21.14 ± 2.96 116.04 ± 9.49 ^*** 69.53 ± 5.13 ^###

Blood leptin content was significantly increased in the high-fat diet control group (Group 2) compared to the control group (Group 1), and significantly decreased in the mdB-44 administration group (Group 3) compared to Group 2. Blood adiponectin content was significantly decreased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and compared to the high-fat diet control group (Group 2), mdB-44 administration group (Group 3) increased. It is known that the increase in the size, number, and quantity of adipocytes in visceral adipose tissue has a positive correlation with the amount of leptin secretion, and a negative correlation with the amount of adiponectin secretion. It significantly reduced the level of leptin in the blood and increased the secretion of adiponectin, thereby exhibiting an anti-obesity effect.

6.6 Histological analysis of epididymal adipose tissue

Fixed epididymal adipose tissue excised from the experimental animal in Example 6.1 was embedded in paraffin, and 5 μm tissue sections were prepared from the embedded tissues. After deparaffinization, tissues were hydrated by sequentially lowering the percentage of alcohol, starting with 100% alcohol to 0% alcohol (H ₂ O). Subsequently, tissues were stained using Accustain® Hematoxylin and Eosin Stains (Sigma-Aldrich Co.) according to the method suggested by the manufacturer (hematocylin & eosin staining, H&E staining), and each tissue was examined using an optical microscope (Carl Zeiss). Histological changes were observed. In addition, the size and number of adipocytes were measured using the AxioVision Imaging analysis system for epididymal adipose tissue pictures obtained by H&E staining. The histological changes of the epididymal adipose tissue are shown in FIG. 3, and the number of cells according to the size of the adipocytes in the measured adipose tissue is shown in Table 14.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 <80μm 161.0 ± 7.1 1.0 ± 1.0 ^*** 41.7 ± 2.2 ^### 80~100μm 16.0±1.7 9.3±1.5 ^* 45.0 ± 3.5 ^### 100~120μm 0.0 ± 0.0 28.0±4.0 ^* 12.7±1.2 ^# >120μm 0.0 ± 0.0 12.7±1.2 ^** 0.0±0.0 ^## equal area fat cells 177.0 ± 8.7 51.0 ± 5.7 ^*** 99.3±5.2 ^## Mean fat globules size (μm) 80.9 ± 0.1 108.5 ± 1.0 ^*** 88.3 ± 0.4 ^###

(The above figures are expressed as mean ± standard error, and mean ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 )

As shown in Figure 3, compared to the control group (Group 1), the size of adipocytes in the epididymal adipose tissue of the high-fat diet control group (Group 2) was significantly increased, and in the mdB-44 administration group (Group 3), adipocytes showed a decreasing trend in size.

In addition, as shown in Table 14, which shows the results of measuring the number of cells according to the size of adipocytes in epididymal adipose tissue, the control group (group 1) had 161.0 ± 7.1 cells with a size of <80 μm, whereas the number of cells with a size of <80 μm was 161.0 ± 7.1. The number of adipocytes >100 μm was not observed, confirming that the size of adipocytes was generally small. The number of cells with a size of 100-120 μm in the high-fat diet control group (group 2) was 28.0 ± 4.0, and the number of cells with a size >120 μm was 12.0 ± 2.1, confirming that the size of the fat cells was large. Compared to the high-fat diet control group (Group 2), the number of cells by size of adipocytes in the test substance-administered group (Group 3) was compared. , the number of cells 100-120 μm in size was significantly decreased. The number of >120μm adipocytes was not observed in the mdB-44 200mg/kg BW administered group (Group 3).

The total number of adipocytes compared to the same area was significantly reduced to 51.0 ± 5.7 in the high-fat diet control group (Group 2) compared to 177.0 ± 8.7 in the control group (Group 1), and the mdB-44 200mg/kg BW administration group (Group 2) 3) increased significantly. The average fat globule size was 108.5 ± 1.0 µm in the high-fat diet control group (Group 2) compared to 80.9 ± 0.1 µm in the control group (Group 1), and the fat globule size significantly increased, mdB-44 200mg/kg In the BW-administered group (Group 3), the size of fat globules was significantly reduced to 88.3 ± 0.4 μm (Group 3).

From the above results, it was confirmed that the administration of the quince extract not only exhibits an anti-obesity effect, but also has an effect of suppressing hypertrophy of white adipose tissue caused by a high-fat diet.

6.6 Analysis of mRNA expression of genes related to energy metabolism in adipose tissue

Total RNA was isolated using TRIzol® Reagent (Thermo Fisher Scientific) from the epididymal adipose tissue obtained in Example 6.1, and then quantified using a micro-volume spectrophotometer (BioSpec-nano, SHIMADZU), and the OD260/280 value was 1.8. The above RNA was used in the experiment. After obtaining cDNA from total RNA (2 μg) using HyperScriptTM RT master mix kit (GeneAll Biotechnology), real-time PCR using Rotor-Gene 300 PCR (Corbett Research) and Rotor-GeneTM SYBR Green kit (QIAGEN) was performed. Primer information used in the experiment is shown in Table 15. Quantitative analysis of gene expression was performed using the Rotor-Gene 6000 Series System Software program (Corbett Research).

mRNA Base sequence (5' → 3') Genebank no. sequence number C/EBPa-FOR primer TGGACAAGAACAGCAACGAGTAC XM_021168520.2 One C/EBPa-REV primer GCAGTTGCCCATGGCCTTGAC XM_021168520.2 2 PPARγ-FOR primer CAAAACACCAGTGTGAATTA XM_021164279.2 3 PPARγ-REV primer ACCATGGTAATTTTCTTGTGA XM_021164279.2 4 SREBP-1c-FOR primer CACTTCTGGAGACATCGCAAAC NM_011480.4 5 SREBP-1c-REV primer ATGGTAGACAACAGCCGCATC NM_011480.4 6 CPT1β-FOR primer GTGCTGGAGGTGGCTTTGGT NM_009948.2 7 CPT1β-REV primer TGCTTGACGGATGTGGTTCC NM_009948.2 8 FAS-FOR primers AGGGGTCGACCTGGTCCTCA NM_007988.3 9 FAS-REV primers GCCATGCCCAGAGGGTGGTT NM_007988.3 10 aP2-FOR primer GGATTTGGTCACCATCCGGT NM_024406.3 11 aP2-REV primer TTCACCTTCCTGTCGTCTGC NM_024406.3 12 HSL-FOR primers CCGTTCCTGCAGACTCTCTC XM_030242181.1 13 HSL-REV primers CCACGCAACTCTGGGTCTAT XM_030242181.1 14 GAPDH-FOR primer AGTTGTTCTCCTGCGACT XM_029478683.1 15 GAPDH-REV primer TGCTGTAGCCGTATTCATTGTCA XM_029478683.1 16 ACC1-FOR primer GGAGATGTACGCTGACCGAGAA AY451393.1 17 ACC1-REV primer ACCCGACGCATGGTTTTCA AY451393.1 18 ACL-FOR primers TGGATGCCACAGCTGACTAC BC056378.1 19 ACL-REV primer GGTTCAGCAAGGTCAGCTTC BC056378.1 20

The effect of the test substance on the expression of mRNA related to fat synthesis, degradation and energy metabolism in epididymal adipose tissue was analyzed, and the results are shown in Table 16.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 C/EBPα 1.00 ± 0.36 4.10 ± 0.05 ^*** 2.20±0.48 ^## PPARγ 1.00 ± 0.12 2.25±0.20 ^** 1.11±0.06 ^## SREBP-1c 1.00 ± 0.12 2.41±0.23 ^** 1.51±0.17 ^# CPT1 1.00 ± 0.19 0.37±0.07 ^* 2.28 ± 0.14 ^### FAS 1.00 ± 0.52 2.90 ± 0.39 ^* 0.25 ± 0.10 ^### aP2 1.00 ± 0.11 1.98±0.38 ^* 0.94±0.13 ^# HSL 1.00 ± 0.07 0.45±0.04 ^*** 0.78±0.07 ^## ACC1 1.01 ± 0.08 3.66±0.42 ^*** 2.82 ± 0.35 ACL 0.96 ± 0.14 1.49±0.15 ^* 0.75±0.16 ^#

(The above results were calculated as the expression amount of each substance compared to the amount of GAPDH expression, (the expression amount of each substance / the expression amount of GAPDH), and were expressed as mean ± standard error, ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001)

Differentiation from pre-adipocytes to mature adipocytes takes place only when transcriptional activators are activated in the regulatory regions of adipocyte genes. Activated Receptor) and C/EBPs (CCAAT/enhancer binding protein). SREBPs include SREBP-1 and SREBP-2, among which SREBP-1 is rapidly induced in the early stage of preadipocyte differentiation, promotes adipocyte differentiation, and promotes fat metabolism by increasing the expression of genes related to fat metabolism. do.

As shown in Table 16, SREBP-1c mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and the high-fat diet control group (Group 2) Compared to the mdB-44 200mg / kg BW administration group (group 3) significantly decreased. C/EBPβ is expressed by hormone induction (insulin, dexamethasone, etc.) in the early stage of adipocyte differentiation, and when C/EBPβ is activated, the expression of C/EBPα and PPARγ, transcription factors of adipocytes, is induced in the late stage of differentiation. C/EBPα has an effect by interacting with PPARγ. C/EBPα increases before the expression of specific genes in adipose tissue to regulate energy homeostasis. PPARγ is a major regulator of adipogenesis and is highly expressed in white adipose tissue. C/EBPα and PPAR-γ mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and compared to the high-fat diet control group (Group 2). It was significantly decreased in the mdB-44 200mg/kg BW administration group (Group 3).

Fatty acid synthase (FAS) is an enzyme involved in the biosynthesis of fatty acids. When fatty acids are synthesized in the liver, they are converted to triglycerides and transport them to adipose tissue in the form of VLDL, circulating blood vessels. FAS mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and mdB-44 200mg/kg BW compared to the high-fat diet control group (Group 2) It was significantly decreased in the administration group (group 3).

Known as a target gene for fat synthesis, aP2 is expressed in adipocytes and macrophages and is known to be involved in inflammation and metabolic reactions. The aP2 mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and mdB-44 200mg/kg compared to the high-fat diet control group (Group 2). In the BW administration group (Group 3), aP2 mRNA expression was significantly decreased (Table 16).

CPT-1 (Carnitine palmitoyl transferase-1) is a rate-limiting step enzyme that imports fatty acids into mitochondria. In order for fatty acids to enter cells and be oxidized, membrane transferase is required. play a major role in its use. CPT-1 mRNA expression in epididymal adipose tissue was significantly decreased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and CPT-1 mRNA expression increased in the mdB-44 administration group (Group 3). High-fat diet was significantly increased compared to the control group (Group 2).

HSL (hormone-sensitive lipase) is distributed in various organs, but is mainly activated in adipose tissue, and becomes active HSL by secretion of hormones such as epinephrine and glucagon. It breaks down into fatty acids. HSL mRNA expression in epididymal adipose tissue was significantly decreased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1). Compared to the high-fat diet control group (Group 2), the HSL mRNA expression significantly increased in the mdB-44 200mg/kg BW group (Group 3) (Table 16).

ACC1 is mainly present in adipose tissue where fatty acid synthesis is important, and ACC2 is mainly present in oxidative tissues such as skeletal muscle and heart to regulate fatty acid synthesis and oxidation. ACC1 mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and mdB-44 200mg/kg compared to the high-fat diet control group (Group 2) ACC1 mRNA expression decreased in the BW administration group (Group 3)

ACL is an enzyme involved in the synthesis of cytosolic acetyl-CoA by converting citrate to acetyl-CoA, and is used in important biosynthetic pathways such as fatty acids and cholesterol. ACL mRNA expression in epididymal adipose tissue was significantly increased in the high-fat diet control group (Group 2) compared to the control diet group (Group 1), and mdB-44 200mg/kg compared to the high-fat diet control group (Group 2). ACL mRNA expression was significantly decreased in the BW administration group (Group 3).

6.7 Analysis of protein expression in adipose tissue

To investigate protein expression in epididymal adipose tissue, lysis buffer (Pierce® IP Lysis buffer, Thermo Scientific) was added to epididymal adipose tissue and then homogenized with a homogenizer. The homogenized solution was centrifuged at 12,000 rpm for 10 minutes, and the supernatant was taken to obtain epididymal adipose tissue lysates. The amount of protein in the tissue lysate was measured using the BCA protein assay kit (Thermo Scientific).

Protein (50μg) was separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride membrane (Milipore). After blocking the membrane in 5% skim milk-TBST (20 mmol/L Tris HCl, pH 7.5, 150 mmol/L NaCl, 0.1% Tween 20) for 1 hour, each antibody to be measured was added to the temperature at 4°C. Stir for 16 hours or at room temperature for 1 hour. Antibody information used in the experiment is shown in Table 17. After that, horseradish peroxidase (HRP)-linked anti-rabbit IgG or HRP-linked anti-mouse IgG was added and stirred for 1 hour, and each protein band was visualized by enhanced chemiluminscence method using LuminataTM Forte Western HRP Substrate (Millipore). . Protein expression levels were quantified using ImageQuantTM LAS 500 imaging systems (GE Healthcare Bio-Sciences AB).

antibody name details manufacture company p-AMPK Phospho-AMPKα (Thr172) #2535 Cell Signaling Technology AMPK AMPKα #2532 Cell Signaling Technology β-actin Beta-actin #3700 Cell Signaling Technology

AMP-activated protein kinase (AMPK) is a factor that improves insulin resistance. Activated AMPK is known to activate ATP production pathways such as glycolysis and fatty acid oxidation while blocking ATP consumption pathways such as fat and cholesterol synthesis. . In particular, it has been reported to play an important role in reducing body fat due to increased fatty acid oxidation by inhibiting the activity of acetyl-CoA carboxylase (ACC) enzyme through protein phosphorylation in relation to lipid metabolism.

In order to investigate the effect of the test substance on AMPK activity in epididymal adipose tissue, Western blot was performed and quantified using epididymal adipose tissue lysate, and the results are shown in Table 18.

group 1 group 2 group 3 empirical diet CD HFD HFD Test substance (mg/kg BW) - - mdB-44 200 p-AMPK/AMPK Ratio 1.00 ± 0.0 0.64 ± 0.04 ^* 1.50±0.22 ^#

(The above figures are expressed as mean ± standard error, and mean ^* p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001 )

As shown in Table 18, as a result of evaluating AMPK activity through the ratio of p-AMPK / AMPK, the activity of AMPK decreased in the high-fat diet control group (Group 2) was found in the mdB-44 200mg / kg BW administration group (Group 3). increased significantly.

As confirmed above, the quince extract administered with the high-fat diet effectively suppressed the increase in body weight, liver and adipose tissue weight, blood cholesterol, blood insulin and leptin content induced by the high-fat diet, and It was confirmed that the size increase was inhibited. This suggests that quince has anti-obesity and lipid-improving effects on dietary obesity, and further has preventive, therapeutic, and/or ameliorative effects on metabolic diseases, and that it has future anti-obesity effects and/or preventive, It suggests the possibility of development as a health functional material having therapeutic and/or improving effects.

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Claims (18)

A food composition for lowering blood sugar or blood cholesterol, comprising a quince extract. According to claim 1, wherein the quince extract is an extract obtained by extracting quince with at least one extraction solvent selected from the group consisting of water and C1-C4 straight-chain or branched-chain alcohol. The method of claim 1, wherein the quince extract is
An extract obtained by extracting quince with 10 to 70 (v/v)% alcohol,
A food composition for lowering blood sugar or blood cholesterol. The food composition for lowering blood sugar or blood cholesterol according to any one of claims 1 to 3, wherein the quince is in the form of a solid remaining after removing the quince juice after pressing. The food composition according to claim 2 or 3, wherein the alcohol extraction is performed by adding 1 to 10 times the volume of alcohol at 50 to 100 ° C and extracting for 1 to 24 hours. According to any one of claims 1 to 3, wherein the extract has a sugar content of 10 to 100 brix, blood sugar or blood cholesterol lowering food composition. According to any one of claims 1 to 3, wherein the food is a health functional food, blood sugar or blood cholesterol food composition for lowering. The food composition for lowering blood sugar or blood cholesterol according to any one of claims 1 to 3, which has an effect of preventing or improving metabolic diseases. According to claim 8, wherein the metabolic disease is diabetes, hyperlipidemia, arteriosclerosis, hyperinsulinemia or metabolic syndrome, blood sugar or blood cholesterol lowering food composition. A food composition for preventing or improving metabolic diseases, comprising a quince extract. 11. The method of claim 10, wherein the quince extract is an extract obtained by extracting quince with one or more extraction solvents selected from the group consisting of water and C1-C4 straight-chain or branched-chain alcohol. 11. The method of claim 10, wherein the quince extract is
An extract obtained by extracting quince with 10 to 70 (v/v)% alcohol,
A food composition for preventing or improving metabolic diseases. The food composition for preventing or improving metabolic diseases according to any one of claims 10 to 12, wherein the quince is in the form of a solid remaining after removing the quince juice after pressing. The food composition according to claim 11 or 12, wherein the alcohol extraction is performed by adding 1 to 10 volumes of alcohol at 50 to 100 ° C and extracting for 1 to 24 hours. According to any one of claims 10 to 12, wherein the extract has a sugar content of 10 to 100 brix, metabolic disease preventing or improving food composition. According to any one of claims 10 to 12, wherein the food is a health functional food, a food composition for preventing or improving metabolic diseases. The food composition for preventing or improving metabolic diseases according to any one of claims 10 to 12, which has an effect of lowering blood sugar or blood cholesterol. According to claim 10 to claim 12, wherein the metabolic disease is diabetes, hyperlipidemia, arteriosclerosis, hyperinsulinemia or metabolic syndrome, metabolic disease preventing or improving food composition.

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