KR101890853B1 - A composition for prevention or treatment of obesity comprising protamine and chitooligosaccharide - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating obesity comprising protamine and chito oligosaccharide as an active ingredient, and a food composition for preventing or ameliorating obesity comprising protamine and chito oligosaccharide as an active ingredient.

Description

TECHNICAL FIELD The present invention relates to a composition for prevention or treatment of obesity comprising protamine and chito oligosaccharide,

The present invention relates to a pharmaceutical composition for preventing or treating obesity comprising protamine and chito oligosaccharide as an active ingredient, and a food composition for preventing or ameliorating obesity comprising protamine and chito oligosaccharide as an active ingredient.

In recent years, as the national income has improved along with the economic growth and the eating habits have become westernized, the intake of various animal foods and processed foods is increasing. This diet leads to overweight. In fact, WHO (World Health Organization), in 2005, 54% of Korean women and 45% of men are overweight, and by 2015, 67% of women and 66% of men . One of the reasons for the increase in overweight is the westernized diet and the traditional social culture of Korea. In other words, with the development of trade and traffic, western foods and western food culture became widely known, making it possible to easily access high-fat foods such as fast food and bread. In addition, it is frequent to drink high fat foods such as meat through drinking, which is one of the traditional social cultures of Korea. While eating these high-fat foods frequently, fat is constantly accumulating in the human body due to lack of exercise. Excessively accumulated fat leads to overweight, and overweight develops into obesity that causes diseases such as cardiovascular disease, cancer, hyperlipidemia, diabetes, sexual dysfunction, and arthritis. Obesity is not only a problem of personal habits but also a social problem and a global health problem. For this reason, many efforts have been made to prevent and treat obesity.

Pancreatic lipase (pancreatic lipase) is an enzyme that is very important for the absorption of fat, hydrolysis of the body into the body to absorb the role of the body. Fats hydrolyzed by pancreatic lipase migrate to the liver and circulate and accumulate throughout the body in the form of triglycerides and cholesterol. In this regard, much research has been conducted to find a substance having an anti-obesity effect by inhibiting pancreatic lipase.

Protamine is a basic protein that exists in the mature sperm nucleus of vertebrates. Protamine is used as an antidote to heparin. It is also used as a food preservative because of its strong antibacterial effect. . The protamine is known to inhibit absorption of ingested fat, thereby reducing fat accumulation and further reducing body weight.

Chitooligosaccharide is an oligosaccharide obtained by hydrolyzing polychitosan. Recently, chitooligosaccharide has attracted attention as a new biomedical material because of its good solubility in water and various therapeutic effects. Among the therapeutic effects of chitooligosaccharide, it is possible to reduce the intake of antidiabetic drugs and to prevent the ingestion of anti-cancer, anti-bacterial, anti-fungal, antioxidant and anti- There is anti-mutagenic. As a health food, chitooligosaccharide is known to restore blood pressure, improve immune function and prevent liver disease caused by alcohol.

Accordingly, the present inventors have made intensive efforts to develop a composition having an anti-obesity effect. As a result, the present inventors have found that a composition comprising protamine and chito oligosaccharide as an effective ingredient effectively inhibits pancreatic lipase activity and inhibits triglyceride and LDL-cholesterol And the level of HDL-cholesterol was increased to confirm the effect of preventing or treating obesity, and the optimum weight ratio of protamine and chito oligosaccharide was determined to complete the present invention.

The present invention provides a pharmaceutical composition for preventing or treating obesity comprising protamine and chito oligosaccharide as an active ingredient.

The present invention also provides a food composition for preventing or ameliorating obesity comprising protamine and chito oligosaccharide as an active ingredient.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating obesity comprising protamine and chito oligosaccharide as an active ingredient.

The term "protamine " in the present invention means a basic protein derived from Oncorhynchus testis, which has a small molecular weight of 4,000 to 5,000 and serves to prevent DNA damage. For the purpose of the present invention, the protamine may mean a substance having an anti-obesity effect, preferably a substance inhibiting the enzyme activity of pancreatic lipase. The protamine of the present invention can be obtained by digesting a basic protein bound to a nucleic acid in a testis such as Oncorhynchus, Clupea , Scomber, Cyprinus, and Oncorhynchus into an acid and then neutralizing it . The protamine may be obtained by neutralizing the raw material with hydrolysis under hydrothermal conditions, followed by salting out to obtain a protein component of protamine, and may be used in powder form.

The term "chitooligosaccharide " in the present invention means an oligosaccharide obtained by hydrolyzing polychitosan. The chitooligosaccharide has a small molecular weight and is characterized by good absorbability within the human body and low viscosity. For the purpose of the present invention, the chitooligosaccharide may mean an oligosaccharide having an anti-obesity effect, preferably a substance inhibiting the enzyme activity of pancreatic lipase.

In the present invention, chitooligosaccharide is deacetylated with chitin obtained by deproteinization and decalcification of bones of crustaceans (crabs, shrimp, etc.), mollusk (squid, squid, etc.) according to a known method, Hydrolyzed by enzymatic treatment can be used.

The term "pharmaceutical composition for the prevention or treatment of obesity" in the present invention means an effect of at least one of weight loss, reduction of blood lipid concentration, reduction of liver tissue and body fat, reduction of glucose concentration in plasma, improvement of liver lipid, and inhibition of leptin secretion And the composition can be used to improve, prevent, or treat various metabolic diseases such as diabetes, hyperlipidemia, heart disease, stroke, arteriosclerosis, fatty liver, etc. derived from obesity-related diseases such as obesity and obesity. For the purpose of the present invention, the composition means a composition comprising protamine and chito oligosaccharide as an active ingredient. Diet drink foods and health functional foods each containing protamine or chito oligosaccharide have been commercialized recently, but a composition comprising the mixture of the above two components at a certain ratio as an active ingredient has not been known.

The protamine and chito oligosaccharide in the composition according to the present invention may be included at various weight ratios which can cause synergistic action with each other. Since each component exhibiting an anti-obesity function has a specific mechanism of action, when various components are mixed and acted at a certain ratio before the action, there may be an antagonistic or synergistic action, and it is necessary to find a suitable ratio for the synergistic effect.

The protamine and chito oligosaccharide may be contained in a weight ratio of preferably 5 to 92: 8 to 95, more preferably 5 to 88: 12 to 60, and still more preferably 1: 3 to 1 : 12, and more preferably 1: 6 by weight, but the present invention is not limited thereto.

It is difficult to expect an anti-obesity effect due to mutual synergistic effect when the minimum recommended amount of each ingredient is used. In case of using the amount exceeding the recommended maximum use amount of each ingredient, the effect of the anti-obesity is not directly increased May not be suitable.

In one embodiment of the present invention, protamine and chito oligosaccharide were mixed at a weight ratio of 1: 3 to 1:12 (examples), and the inhibitory activity of pancreatic lipase was measured. As a result, It was confirmed that the effect was most excellent when the weight ratio was 1: 6 to 1: 8 (Experimental Example 1). In addition, the composition containing protamine and chito oligosaccharide as active ingredients showed a much lower level of triglyceride and LDL-cholesterol in the blood of rats fed with fat compared to the case of using protamine or chito oligosaccharide, respectively, while the HDL-cholesterol level (Experimental Example 2).

The mixture of protamine and chitooligosaccharide at a certain ratio is more effective in inhibiting lipidase inhibitory action and absorption of fat in the body than when each of the above components is used alone. Therefore, when the above protamine and chito oligosaccharide are used as an active ingredient Is useful as an anti-obesity composition. Since protamine and chito oligosaccharide are natural physiologically active substances, unlike conventional lipase inhibitors such as orlistat, protamine and chito oligosaccharide are safe and can be used without problems even in large doses because there is no fear of side effects or toxicity when ingested have.

The term "obesity " in the present invention means a state in which the fat tissue is excessively accumulated in the body. The obesity includes a case where the body mass index (BMI) is 25 or more.

The term " prevention "in the present invention means all the actions of suppressing or delaying obesity by the administration of the composition, and" treatment "means that all the actions it means.

The pharmaceutical composition comprising protamine and chito oligosaccharide as an active ingredient may have a total content of protamine and chito oligosaccharide of 0.2% by weight to 100% by weight based on the total weight of the composition.

The pharmaceutical composition may comprise a pharmaceutically acceptable carrier. The composition comprising a pharmaceutically acceptable carrier can be of various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical composition may be in the form of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories It can have one formulation.

The pharmaceutical compositions may be administered by non-oral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal routes, and for localized immunosuppressive treatment, by a suitable method, including, if necessary, by intralesional administration. Non-oral injections include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Preferred modes of administration are intravenous, subcutaneous, intradermal, intramuscular, and drip.

The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. The composition of the present invention is preferably administered in divided doses of 1 to 500 mg, preferably 50 to 100 mg per body weight per day, preferably 1 to 3 times per day in the case of oral administration, The dosage may be administered in a dose of 0.5 mg to 250 mg, preferably 1 to 3 times per 1 kg of body weight, but is not limited thereto. In addition, the composition of the present invention can be used alone or in combination with the methods of using surgery, hormone therapy, drug therapy and biological response modifiers for the prevention or treatment of obesity.

In one embodiment of the present invention, a composition comprising protamine and chito oligosaccharide was prepared on the basis of a lipid complex (Experimental Example 2-4). As a result of administering the composition to rats, triglyceride and LDL-cholesterol And increased the level of HDL-cholesterol. Thus, it was confirmed that the composition of the present invention can be used for prevention or treatment of obesity (Experimental Example 2). In addition, administration of a composition comprising protamine and chito oligosaccharide at a ratio of 1: 6 showed improved effects on body weight, body fat, triglyceride, total cholesterol, and HDL cholesterol as compared with the placebo control group. It was found that the composition containing both components was effective in improving obesity. From these results, it was confirmed that the composition of the present invention can be used for prevention or treatment of obesity (Experimental Example 3).

The present invention also provides a food composition for preventing or ameliorating obesity comprising protamine and chito oligosaccharide as an active ingredient.

The protamine and chito oligosaccharide are as described above. More specifically, the mixture of protamine and chito oligosaccharide of the present invention may be added to the food composition for the purpose of preventing or improving obesity.

When the protamine and chito oligosaccharide of the present invention are used as a food additive, the protamine and chito oligosaccharide can be directly added or used together with other foods or ingredients, and they can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use. Generally, the total content of protamine and chito oligosaccharide in the manufacture of food or beverage can be adjusted to 0.2 to 100% by weight based on the total weight of the food composition. However, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the amount can also be used in the above-mentioned range.

The kind of the food of the present invention is not particularly limited. Examples of foods to which the protamine and chito oligosaccharide can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen and other noodles, gums and ice cream, health functional foods, , Beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and may include all foods in a conventional sense, and foods used as feed for animals. The content of the protamine and chito oligosaccharide according to the present invention in the composition of the present invention is not particularly limited as long as the content of the protamine, And can be adjusted to 0.2 to 100% by weight based on the total weight. There are no particular restrictions on other components other than the protamine and chito oligosaccharide according to the present invention as an active ingredient, and may include various conventional flavors or natural carbohydrates as an additional ingredient.

In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but can be selected in the range of 0 to 20 parts by weight per 100 parts by weight of the composition of the present invention. These components can be used independently or in combination. As an example of the composition, ordinary soft capsules were prepared by mixing the protamine and chitooligosaccharide main components with soybean oil, beeswax (yellow lead), lecithin, etc. commonly used as excipients, and then injecting the mixture into the coating of the gelatin base Production Example 1).

The composition of the present invention comprising protamine and chito oligosaccharide in a certain ratio includes a natural physiologically active substance and can be safely used without adverse effects on the human body unlike a conventional lipase inhibitor. In addition, by containing protamine and chiroooligosaccharide in an appropriate ratio, it effectively inhibits the body lipase and inhibits excessive fat absorption, thereby lowering the body's neutral lipids, total cholesterol, LDL-cholesterol and increasing HDL cholesterol. Can be effectively used for the treatment of obesity and the like.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example : Protamine  And Chitosaurus  Preparation of mixtures

Protamine and chito oligosaccharides were mixed according to the weight ratios shown in Table 1 below to prepare the mixture shown in Table 1 below. Protamine and chitooligosaccharide were purchased and used.

Protamine Chitooligosaccharide Example 1 One 0 Example 2 One 3 Example 3 One 5 Example 4 One 6 Example 5 One 8 Example 6 One 12 Example 7 0 One Control group 0 0

Experimental Example  One: Protamine  And Chitooligosaccharide  Inhibition of lipid-degrading enzymes of the mixture

In order to confirm whether the above Examples 1 to 7 and the control group had an effect on lipase inhibition, the following experiment was carried out.

Measurement of pancreatic lipase inhibitory activity is a method of evaluating how much lipid ingested is hydrolyzed to fatty acids such as triglyceride and then prevented from being absorbed in the intestines. In other words, when the pancreatic lipase is inhibited, it is prevented from being hydrolyzed by fatty acid to be absorbed into the body through the intestines. Therefore, it can be deduced that a large pancreatic lipase inhibiting activity has a good effect on obesity. The pancreatic lipase used in the experiment was porcine pancreatic lipase (Sigma-Aldrich), and 4-MU (4-methylumbelliferyl oleate, Sigma-Aldrich) was used as the substrate of the lipase.

Specifically, 5 μL each of the above Examples 1 to 7 and the control group was added to a 96-well plate, and 0.1 mM 4-MU solution (0.1 mM 4-MU, 13 mM Tris-HCl, 150 mM NaCl, and 1.3 mM CaCl ₂ , pH 8.0) and 25 μL of porcine pancreatic lipase were added thereto, followed by incubation at 37 ° C. for 30 minutes. After incubation, 0.1 M sodium citrate (pH 4.2) was added to stop the reaction. Absorbance was measured at a wavelength of 450 nm using an ELISA reader (VERSA man, Molecular Devices, Sunnyvale, Calif., USA), and the results are shown in Table 2 below. The lipase activity of the following Table 2 indicates the relative enzyme activity in% of the control group as 100%.

Lipase activity (%) Example 1 (1: 0) 60 Example 2 (1: 3) 53 Example 3 (1: 5) 50 Example 4 (1: 6) 40 Example 5 (1: 8) 43 Example 6 (1:12) 48 Example 7 (0: 1) 70 Control group 100

As shown in Table 2, the lipase activity was superior to that of protamine or chito oligosaccharide when protamine or chito oligosaccharide was used alone. When the ratio of protamine to chitooligosaccharide was 1: 3 to 1: 6 (Examples 2 to 4), the lipase activity was continuously decreased and the chitooligosaccharide ratio was recovered to some extent, , It was shown that the lipase-inhibiting effect was superior to the case where the two components were mixed at a total weight ratio of 1: 3 to 1:12.

Experimental Example  2: Protamine  And Chitooligosaccharide  Confirmation of inhibition of fat absorption of mixture Animal room Hum

2-1: Preparation of materials for animal experiments

The lipid complexes orally administered with the test substance were prepared with corn oil, cholic acid, cholesteryloleate and margarine. Corn oil, cholic acid and cholesterylate were all available from Sigma-Aldrich, and margarine from Seoul Milk Co., Ltd. was used. Corn oil is composed of oleic acid, linoleic acid, palmitic acid, etc. These fatty acids are contained in meat fat such as pork belly. Cholinic acid acts to emulsify lipids with bile, and cholesterylate refers to cholesterol and esters of oleic acid. Margarine contains trans fatty acids. In this study, a fat complex was selected for this reason. It can be assumed that when the meat is consumed by containing corn oil, it can be assumed that cholesterol-containing food is ingested by adding cholesterylate thereto. In addition, the effects of protamine and chito oligosaccharides when various lipids were ingested by adding a small amount of margarine could be expected.

2-2: Preparation of experimental animals

Seven-week-old male SD rats were used as experimental animals. This was tested in 8-week-old SD-rats after an adaptation period of one week in an animal laboratory of a conventional area system. Each rat was housed four per cage and raised in an environment controlled at a temperature of 23.0 ± 1 ° C, a humidity of 40 ± 60%, and an illumination of 150 to 300 Lux. During the breeding, all experimental animals were allowed to eat solid feed freely, and the tap water was ingested in a negative water.

2-3: Dietary regulation  And fasting step

Twelve hours before the start of the experiment, we entered the fasting period except for the negative. As the experiment began, 12 g of feed per serving was provided. In this experiment, animals were fed three days prior to the start of the experiment. Three days before the experiment, the feed was fed at a rate of 60 g per cage. During the experiment, one cage was separated and fed with 12 g of feed per 24 hours for the purpose of reducing the error between animals.

2-4: Experimental diet  Produce

Experimental diets were prepared by adding protamine and chito oligosaccharide to each concentration. The lipid complex consisted of 6 ml of corn oil, 80 mg of choline, 2 mg of cholesterylate, 1 mg of margarine, and 6 ml of saline solution. Protamine and chito oligosaccharide were measured in the lipid complexes prepared in the above manner and then the amount of protamine and chito oligosaccharide was measured in each concentration and stored in the tube.

2-5: Serum Triglyceride ( triglyceride ) And cholesterol ( total -cholesterol measurement

Blood was collected from the tail vein of SD-rats to measure serum triglyceride and cholesterol. At this time, 700 cc of blood was collected from the tail vein 0, 3, 9, and 24 hours after the oral administration of the experimental diet. 700 cc of collected blood was placed in a SST tube (Green Separation Tube), left at room temperature for 1 to 2 hours, and centrifuged at 3000 rpm and 4 ° C for 20 minutes. Then, the supernatant serum was separated, and the separated serum was stored at -20 ° C. Then, blood triglyceride, total-cholesterol, high density lipoprotein (HDL) -cholesterol, and LDL (low density lipoprotein) -cholesterol were measured using a blood auto analyzer Hitachi 7080 (Hitachi sciene system Lad.). Were measured. By measuring triglyceride and total cholesterol, we were able to confirm the level of metabolism of orally administered lipid complexes. We found that protamine and / or chitooligosaccharide lowered cholesterol and LDL-cholesterol, which is known to accumulate in the walls of the blood vessels and cause arteriosclerosis , And cholesterol was transported to the terminal tissues and HDL-cholesterol, which prevents accumulation of cholesterol in the blood vessel wall, was increased. The results are shown in Tables 3 to 6 below.

The blood neutral lipid content (%) in the following Table 3 represents the ratio of 100% before consumption.

sample Serum triglyceride content (%) Before ingestion 3 hours 24 hours Example 1 (1: 0) 100 275 150 Example 2 (1: 3) 100 220 120 Example 3 (1: 5) 100 205 90 Example 4 (1: 6) 100 200 50 Example 5 (1: 8) 100 210 86 Example 6 (1:12) 100 210 100 Example 7 (0: 1) 100 250 200 Control group 100 420 250

As shown in Table 3, the blood triglyceride content of the control group was changed to 420% and 250% after 3 hours and 24 hours from the initial level after the fat loading. In the group administered with protamine, chitooligosaccharide or protamine and chitooligosaccharide mixture, The content was lower. Among them, a mixture of protamine and chito oligosaccharide showed a lower content than that of the case of administration of protamine and chito oligosaccharide, respectively. When protamine and chito oligosaccharide are mixed, the effect of preventing the absorption of triglyceride into blood Was better. At this time, when the ratio of protamine and chito oligosaccharide is 1: 3 to 1: 6 (Examples 2 to 4), the blood triglyceride content continues to decrease, and when the chito oligosaccharide mixture ratio further increases, But also at that time, the content was much lower than that of protamine and chitooligosaccharide, respectively.

The total cholesterol content (%) in blood shown in Table 4 below represents the ratio of 100% before consumption.

sample Total cholesterol content in blood (%) Before ingestion 3 hours 24 hours Example 1 (1: 0) 100 120 115 Example 2 (1: 3) 100 115 118 Example 3 (1: 5) 100 105 110 Example 4 (1: 6) 100 80 100 Example 5 (1: 8) 100 100 105 Example 6 (1:12) 100 110 115 Example 7 (0: 1) 100 115 120 Control group 100 140 140

As shown in Table 4, the total cholesterol content of the control group increased to 140% and 140% after 3 hours and 24 hours after the fat loading, respectively. However, in the group administered with protamine, chitooligosaccharide or a mixture of protamine and chitooligosaccharide, The total cholesterol content of the blood was lower than that of the control group.

Among them, a mixture of protamine and chito oligosaccharide shows a lower content than that of each of protamine or chito oligosaccharide, showing that when protamine and chito oligosaccharide are mixed, the blood does not increase the total cholesterol content in blood Indicating that the effect is better. At this time, when the ratio of protamine and chito oligosaccharide is 1: 3 to 1: 6 (Examples 2 to 4), the blood cholesterol content is continuously decreased, and when the chito oligosaccharide mixture ratio is further increased, But it was lower than that of protamine and chitooligosaccharide, respectively. Particularly, when the weight ratio was 1: 6 (Example 4), the lowest total cholesterol content in blood was shown.

The blood LDL cholesterol content (%) in Table 5 below represents the ratio of 100% before consumption.

sample Blood LDL cholesterol content (%) Before ingestion 3 hours 24 hours Example 1 (1: 0) 100 102 110 Example 2 (1: 3) 100 60 90 Example 3 (1: 5) 100 75 85 Example 4 (1: 6) 100 90 80 Example 5 (1: 8) 100 100 94 Example 6 (1:12) 100 125 100 Example 7 (0: 1) 100 103 108 Control group 100 140 130

As shown in Table 5, the blood LDL cholesterol of the control group was changed to 140% and 130% after 3 hours and 24 hours from the initial fat load, respectively. However, in the group administered with protamine and chitooligosaccharide or with a mixture of protamine and chito oligosaccharide, Low LDL cholesterol level in the blood.

In particular, in the case of the mixture of protamine and chito oligosaccharide, the blood LDL cholesterol content was generally lower after 3 hours as compared with the group administered with protamine or chito oligosaccharide. Especially when the ratio was 1: 3 to 1: 6, Low. After 24 hours, a mixture of all protamine and chito oligosaccharides showed lower LDL cholesterol content than the control, Examples 1 and 7. In addition, when the ratio of protamine and chito oligosaccharide was 1: 3 to 1: 6, the content of LDL cholesterol was continuously decreased. When the mixing ratio was further increased, the level of cholesterol in blood also increased to some extent, Which was significantly lower than that of the control group.

The HDL cholesterol content (%) in the blood in Table 6 below represents the ratio when the pre-ingestion was regarded as 100%.

sample Serum HDL cholesterol content (%) Before ingestion 3 hours 24 hours Example 1 (1: 0) 100 120 135 Example 2 (1: 3) 100 130 150 Example 3 (1: 5) 100 123 175 Example 4 (1: 6) 100 110 190 Example 5 (1: 8) 100 137 180 Example 6 (1:12) 100 145 150 Example 7 (0: 1) 100 125 140 Control group 100 115 120

As shown in Table 6, HDL cholesterol in the control group was changed to 115% and 120% after 3 hours and 24 hours after the fat loading, respectively. However, in the group administered with protamine and chitooligosaccharide or with a mixture of protamine and chito oligosaccharide, And showed a tendency to increase with time. In addition, HDL cholesterol content was higher when a mixture of protamine and chito oligosaccharide was administered than when protamine and chito oligosaccharide were respectively administered, and thus it was found that the above effect was better when the above mixture was used. After 24 hours, when the ratio of protamine to chito oligosaccharide is 1: 3 to 1: 6 (Examples 2 to 4), the content of HDL cholesterol in the blood continuously increases, and when the ratio of chito oligosaccharide is increased further, Cholesterol levels also decreased to some extent.

2-6: Animal Fecal Triglyceride ( Triglyceride ) And cholesterol ( Total - cholesterol ) Measure

The feces were collected from the feces (nomal) secreted during one week of adaptation period and the feces after 0, 3, 9, and 24 hours after the start of the experiment. Because 18 - hour fasting and minimum feed were supplied, fecal incidence was not generated much, and feces were collected every hour per group. Fecal samples were collected over 5g per hour. The collected feces were stored at -20 ° C. The stored feces were measured by using a Rose-Gottlieb method to extract crude fat and using a kit capable of measuring triglyceride and cholesterol.

The crude fat was extracted from the feces collected by the above method using the Rösinger-Gotlib method, one of the fat analysis methods. The extracted crude fat was measured using a kit capable of measuring triglyceride and cholesterol. 5 g of the feces was cut into small pieces, and the crude fat was extracted with concentrated ammonia water, 95% alcohol, diethly ether, and petroleum ether. Triglyceride Assay kit (Cayman chemical company) and Enzychrome Cholesterol Assay (Enzychrome) were used for crude fat, and triglyceride and cholesterol were analyzed by each experimental method. The results of the analysis are shown in Tables 7 and 8 below.

The fecal triglyceride in the following Table 7 shows the results when the control group was regarded as 100%, and the total cholesterol in the feces of the following Table 8 shows the relative ratio when the pre-intake was 100%.

sample Fecal triglyceride (%) Before ingestion 24 hours Example 1 (1: 0) 100 200 Example 2 (1: 3) 100 300 Example 3 (1: 5) 100 230 Example 4 (1: 6) 100 200 Example 5 (1: 8) 100 210 Example 6 (1:12) 100 250 Example 7 (0: 1) 100 250 Control group 100 150

sample Total cholesterol in feces (%) Before ingestion 24 hours Example 1 (1: 0) 100 185 Example 2 (1: 3) 100 150 Example 3 (1: 5) 100 130 Example 4 (1: 6) 100 120 Example 5 (1: 8) 100 130 Example 6 (1:12) 100 140 Example 7 (0: 1) 100 130 Control group 100 90

In the above Tables 7 and 8, the triglyceride and cholesterol content in the feces did not show a specific tendency depending on the mixing ratio of protamine and chito oligosaccharide, but showed an increasing tendency compared to the control. Especially in the case of triglyceride, protamine compared to the control. Respectively, and a significant increase in fecal emission during administration of each of the chitooligosaccharides or the combination of protamine and chitooligosaccharide.

Experimental Example  3: Human test  Checking for the improvement of body fat through

In order to confirm the body fat improvement effect of protamine and chito oligosaccharide complex, a human body test was conducted on subjects having body mass index (BMI) of 23 or more, serum triglyceride 1.65 mmol / L and blood cholesterol of 5.2 mmol / L . The mixing ratio of protamine and chito oligosaccharide was determined according to the above Example 4, and the daily intakes of the complex were 500 mg, and the mice were fed with 500 mg tablets per day for 8 weeks. The subjects were divided into two groups: 20 in the intake group and 20 in the placebo control group. The exercise and meals were not controlled.

After 8 weeks, body weight and body fat mass were measured using a body composition analyzer (Inbody) as a measurement index, and blood cholesterol, triglyceride, LDL cholesterol, and HDL cholesterol were analyzed using a blood automation analyzer. It was the same.

sample weight
(kg) Body fat
(kg) Triglyceride
(mmol / L) Total cholesterol
(mmol / L) HDL cholesterol
(mmol / L) Placebo control -1.1 - 1.8 0.14 0.5 0.03 Complex intake group -2.2 - 2.2 -1.5 -0.9 1.05

As shown in Table 9, when the complex was used for 8 weeks, the body weight was reduced to 2.2 kg, body fat was reduced to 2.2 kg, blood triglyceride was decreased to 1.5 mmol / L, total cholesterol was decreased to 0.9 mmol / L, HDL cholesterol was decreased to 1.05 mmol / Respectively. In other words, it was confirmed that protamine and chito oligosaccharide showed the effect of reducing the body weight and body fat by bar.

Therefore, it was confirmed that when a composition containing protamine and chito oligosaccharide according to the present invention is used at a certain ratio, it is possible to effectively prevent or suppress obesity.

Manufacturing example  1: Preparation of soft capsules

Soft capsules were prepared using soybean oil, waxy beeswax (liquor) and lecithin, which contain protamine and chito oligosaccharide as main components and are commonly used in soft capsule formulation, in the contents shown in Table 10 below.

Raw material name Formulation ratio (%) Content (mg) Mixed content
(500 mg)

Protamine 7.1 35.5 Chitooligosaccharide 42.9 214.5 Soybean oil 45.0 225 Beeswax 4.0 20 lecithin 1.0 5 (sum) 100.0 500 Soft capsule mechanism
(185 mg) gelatin 68.850 glycerin 21.862 Manatol 7070 8.197 Ethyl vanillin 0.164 Titanium dioxide 0.344 Food color blue No. 1 0.004 Food color blue No. 3 0.028 Food Color Red No. 40 0.551 sum 100,000

As shown in Table 10, the mixed contents were mixed and mixed well on a stirrer, and then injected into a soft capsule base to prepare a conventional soft capsule. The above Preparation Example is a preparation example for producing a conventional soft capsule containing a composition having the above-mentioned anti-obesity effect, and is not intended to limit the formulations and raw materials of the present invention.

Manufacturing example  2: Preparation of Hard Capsule

Hard capsules were prepared using the excipients conventionally used in hard capsules containing protamine and chito oligosaccharide as main components and the contents as shown in Table 11 below.

Raw material name Formulation ratio (%) Content (mg) Protamine 9.9 49.5 Chitooligosaccharide 59.1 295.5 Crystalline cellulose 27.0 135 Magnesium stearate 1.0 5.0 sum 100 500

As shown in Table 11, the contents of the mixture were mixed and mixed well in an agitator to granulate, and then injected into a hard capsule base to prepare a conventional hard capsule. The above preparation example is a preparation example for producing a conventional hard capsule containing a composition having the above-mentioned anti-obesity effect, and thus does not limit the formulations and raw materials of the present invention.

Manufacturing example  3: Preparation of tablets

Tablets were prepared with the contents as shown in Table 12 below, using a tablet, a sweetener, a flavoring agent, and the like, which contain protamine and chito oligosaccharide as main components and are excipients commonly used in tablets.

Raw material name Formulation ratio (%) Content (mg) Protamine 7.1 35.5 Chitooligosaccharide 42.9 214.5 Xylitol 15.0 75 Lactose 10.0 50 Apple taste powder 14.0 70 Apple fruit powder 9.4 47 HPMC 0.6 3 Magnesium stearate 1.0 5 gun 100 500

As shown in Table 12, the contents of the mixture were mixed and granulated thoroughly in an agitator, followed by injection into a tablet machine to prepare a conventional tablet. The preparation examples are not intended to limit the formulations and raw materials of the present invention to preparations for producing conventional tablets containing the composition having the anti-obesity effect of the present invention.

Manufacturing example  4: Manufacture of beverages

A flavoring agent, a preservative, an acidity adjusting agent, a thickening agent, etc., which contain protamine and chito oligosaccharide as main components and which are commonly used as beverages in drinks,

Raw material name content(%) Content (mg) Protamine 0.7 3.5 Chitooligosaccharide 4.1 20.5 Sweetener (xylitol) 2.0 10 Sweetener (D-sorbitol solution) 1.5 7.5 Flavor (apple) 0.1 0.5 Flavor (fructose) 1.0 5 Preservative (sodium benzoate) 0.06 0.3 Acidity regulator (citric acid) 0.1 0.5 Thickener (gum arabic) 0.2 1.0 Distilled water 90.24 451.2 sum 100.00 500

As shown in Table 13, the contents of the mixture were uniformly mixed in distilled water using a high-speed stirrer, sterilized at high temperature, and put into bottles or cans to prepare ordinary beverages. The above preparation examples are preparative examples for preparing ordinary beverages containing a composition having the above-mentioned anti-obesity effect, and are not intended to limit the formulations and raw materials of the present invention.

Claims (8)

A pharmaceutical composition for preventing or treating obesity comprising protamine and chito oligosaccharide as an active ingredient,
Wherein the weight ratio of protamine and chito oligosaccharide is from 1: 3 to 1:12.
The composition of claim 1, wherein the weight ratio of protamine and chito oligosaccharide is from 1: 3 to 1: 6.
3. The composition of claim 2 wherein the weight ratio of protamine and chito oligosaccharide is 1: 6.
The composition of claim 1, wherein the total content of protamine and chito oligosaccharide is from 0.2% to 100% by weight, based on the total weight of the pharmaceutical composition.
A food composition for preventing or ameliorating obesity comprising protamine and chito oligosaccharide as an active ingredient,
Wherein the weight ratio of protamine and chito oligosaccharide is from 1: 3 to 1:12.
6. The composition of claim 5, wherein the weight ratio of protamine and chito oligosaccharide is from 1: 3 to 1: 6.
7. The composition of claim 6, wherein the weight ratio of protamine and chito oligosaccharide is 1: 6.
6. The composition of claim 5, wherein the total content of protamine and chito oligosaccharide is from 0.2% to 100% by weight, based on the total weight of the food composition.