KR20180113014A - A composition for treating, improving and preventing of diabetes mellitus comprising Gracilariopsis chorda ohmi extract or Gracilariopsis chorda ohmi fraction - Google Patents

Abstract

The present invention relates to a composition for effectively preventing or treating diabetes mellitus comprising a Gracilariopsis chorda Ohmi extract or a Gracilariopsis chorda Ohmi fraction, which can treat or prevent diabetes mellitus to effectively manufacture a competitive medicinal composition and a food composition.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for preventing and treating diabetes mellitus, which comprises, as an active ingredient, a plant extract or fraction thereof. The present invention also relates to a pharmaceutical composition for preventing and treating diabetes mellitus comprising Gracilariopsis chorda ohmium extract or Gracilariopsis chorda ohmi fraction,

The invention more kkosiraegi (Gracilariopsis chorda (Holmes) Ohmi ) extract or fraction thereof as an active ingredient to thereby treat, ameliorate or prevent diabetes.

Diabetes mellitus (diabetes mellitus) is insufficient or insensitive to insulin and metabolism of the carbohydrate metabolism is a disease that is abnormal. Diabetes mellitus is classified as type 1 diabetes (insulin-dependent) and type 2 diabetes (non-insulin dependent) according to the developmental stage. Oral hypoglycemic agents currently used in patients with type 1 and type 2 diabetes include α-glucosidase inhibitors and sulfonylureas And dipeptidyl-peptidase 4 (DPP4) inhibitors. The use of a therapeutic agent for diabetes such as α-glucosidase inhibitors, which have been used clinically, may cause side effects such as abdominal bloating, vomiting and diarrhea in some patients when used over a long period of time. Until now, the cure method of diabetes has not been established, and due to the side effect of drug treatment, there are many limitations on the use of drugs, so much attention has been paid to research for searching for diabetes drugs from natural products.

Furthermore, the prolongation of the current lifespan makes it unavoidable that diabetes progresses to complications. In general, after 10 to 20 years of diabetes, almost all the organs in the body are damaged, causing diabetic retinopathy, diabetic cataract, diabetic nephropathy, diabetic neuropathy, Diabetic neuropathy, heart disease, cancer, and osteoporosis. Therefore, a therapeutic agent for diabetic complications which can prevent, treat or delay diabetic complications from diabetic patients is also considered important.

As part of this research, research has been conducted on the possible use of lignans, polabonoids, alkaloids, and terpenoids derived from medicinal plants to alleviate diverse symptoms of diabetes. The present inventors have made efforts to find a substance having a therapeutic effect on diabetes originating from natural products, and as a result, the effect of treating diabetes was confirmed in a pharmaceutical composition containing an effective ingredient derived from a canine.

Patent Document 1. Korean Patent Publication No. 10-1354167

The present invention has been conceived to solve the above-mentioned problems, and an object of the present invention is to provide a composition capable of treating or preventing diabetes by containing an extract of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

In addition, another object of the present invention is to provide a composition that can treat or prevent diabetes by containing a fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

It is still another object of the present invention to provide a food composition containing an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient to improve or prevent diabetes.

In order to achieve the above object, the present invention provides a pharmaceutical composition for treating or preventing diabetes, which comprises an extract of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

The extract of Gracilariopsis chorda (Holmes) Ohmi can be obtained by immersing a gracilariopsis chorda (Holmes) Ohmi in an extraction solvent at 20 to 26 ° C and then immersing it in an extraction solvent at 50 to 80 ° C. have.

The extract of Gracilariopsis chorda (Holmes) Ohmi) may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The mixed solvent may be 20 to 95% by volume of an aqueous solution of methanol, ethanol, butanol or propanol.

The diabetes may be type 2 diabetes caused by reduction of the pancreas induced by alloxan or type 2 diabetes caused by excessive insulin-induced decrease in pancreatic duct.

To achieve these and other advantages and in accordance with the purpose of the present invention, as embodied and broadly described herein, there is provided a pharmaceutical composition for treating or preventing diabetes, comprising a fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

The Gracilariopsis chorda (Holmes) Ohmi fraction may be obtained by fractionation of Gracilariopsis chorda (Holmes) Ohmi extract with hexane, methylene chloride, ethyl acetate, n-butanol or water.

The Gracilariopsis chorda (Holmes) Ohmi fraction is a hexane fraction obtained by sequential fractionation of Gracilariopsis chorda (Holmes) Ohmi extract with hexane, methylene chloride, ethyl acetate and n-butanol, a methylene chloride fraction And water fractions.

The diabetes may be type 2 diabetes caused by reduction of the pancreas induced by alloxan or type 2 diabetes caused by excessive insulin-induced decrease in pancreatic duct.

The pharmaceutical composition for the treatment or prevention of diabetes may be a liquid preparation, an external preparation for skin or an oral preparation.

In order to accomplish the above-mentioned other objects, the present invention provides a food composition for improving or preventing diabetes, which comprises an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

The diabetes may be type 2 diabetes caused by reduction of the pancreas induced by alloxan or type 2 diabetes caused by excessive insulin-induced decrease in pancreatic duct.

According to another aspect of the present invention, there is provided a method of extracting gracilariopsis chorda (Holmes) Ohmi with ethanol or water to prepare an extract of canine corn.

Sequentially fractionating the above-mentioned dog food ethanol extract with hexane, methylene chloride, ethyl acetate and n-butanol and water; And

Concentrating any one or more selected from the above fractions to obtain a seaweed sprout fraction,

The method of manufacturing a food composition for preventing or improving diabetes mellitus according to any one of claims 1 to 3, wherein the open mouth increases the size of the pancreas and increases intracellular glucose uptake.

The extract or fraction according to the present invention is effective for the prevention or treatment of diabetes mellitus, and is highly effective for treating or preventing diabetes, and thus is effective for manufacturing competitive pharmaceutical compositions and food compositions.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic view showing the entire process of preparing the extracts and fractions of canine corn extract of Example 1 and Examples 3 to 7. Fig.
FIG. 2 shows the results of the diabetic zebrafish model induced by alloxan. In order to confirm the anti-diabetic effect of the diabetic zebrafish model induced by the ethanol extract prepared in Example 1 and the hot-water extract prepared in Example 2, Of the pancreatic islet.
FIG. 3 is a graph showing the results of the diabetic zebra fish model induced by alloxan. In order to confirm the anti-diabetic effect of the diabetic zebra fish model induced by the ethanol extract prepared in Example 1 and the hot water extract prepared in Example 2, Of the pancreatic islet.
FIG. 4 is a graph showing changes in the pancreatic islet size of each group in order to confirm the anti-diabetic effect when the diabetic zebra fish model induced by alloxan was administered for a short period of time, to be.
FIG. 5 is a graph showing the results of measuring the fluorescence intensity of pancreatic islets in each group in order to confirm the antidiabetic effect when short-term administration of the dog cauliflower fractions prepared in Examples 3 to 7 in the aloic acid-induced diabetic zebrafish model to be.
6 is a graph showing the results of an experiment to determine the antidiabetic effect when the long-acting dog extract (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Example 1 and Example 2 was orally administered in the dirogenic zebrafish model induced by alloxan , And pancreatic islet size change of each group.
7 is a graph showing the results of an experiment to determine the antidiabetic effect when the dog extract (1 μg / ml, 10 μg / ml) prepared from Example 1 and Example 2 was orally administered to a diabetic zebrafish model induced by alloxan , And pancreatic islet fluorescence of each group.
8 is a graph showing the results obtained when the dog extract (1 μg / ml, 10 μg / ml) prepared in Examples 3 to 7 was administered for a long period in an allograft-induced diabetic zebrafish model. Fig. 2 is a graph showing changes in pancreatic islet size of the group.
FIG. 9 shows the results obtained when the diabetic zebra fish model induced by alloxan was subjected to long-term administration of the macrocarpal fractions (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Examples 3 to 7, Lt; RTI ID = 0.0 &gt; fluorescence < / RTI &gt;
10 is a graph showing the effect of the extracts of dogs (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Examples 1 and 2 on the diabetic zebrafish model induced by excessive insulin, Fig. 2 is a graph showing changes in size of pancreatic islets.
Fig. 11 shows the effect of the extract of dogs (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Examples 1 and 2 on the diabetic zebrafish model induced by excessive insulin, And the fluorescence intensity of the pancreatic islet is measured.
Figure 12 shows the effect of the diabetic fraction produced from Examples 3 to 7 (1 占 퐂 / ml, 10 占 퐂 / ml) on the diabetic zebra fish model induced by excessive insulin in each group Fig. 2 is a graph showing changes in size of pancreatic islets.
FIG. 13 is a graph showing the anti-diabetic effect of the extract of dogs (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Examples 1 and 2 in a diabetic zebrafish model induced by excessive insulin. And the fluorescence intensity of the pancreatic islet is measured. ^{(## p <0.01, ### p} <0.001; control group and bigyodoem), (* p <0.05, ** p <0.01, *** p <0.001; bigyodoem and untreated control group)

Accordingly, the present inventors have made intensive research to develop a novel diabetes therapeutic agent, and as a result, discovered a pharmaceutical composition for the treatment or prevention of diabetes according to the present invention, and completed the present invention.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to a pharmaceutical composition for treating or preventing diabetes, which comprises an extract of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

It is called Gracilariopsis chorda (Holmes) Ohmi . It is native to Korea, China, and Japan. Especially, it is found in the southern coastal waters of Korea.

The open kkosiraegi (Gracilariopsis chorda (Holmes) Ohmi) extracts one kkosiraegi (Gracilariopsis chorda (Holmes) Ohmi) and then immersed in the extraction solvent of 20 to 26 ℃ by immersion in the extraction solvent of 50 to 80 ℃ be obtained extract have. When the extraction conditions for obtaining the extract are out of the above range, the effective amount of the extract can be extracted in a small amount.

For example, the extracts of the present invention are immersed in an extraction solvent at 20 to 26 캜, cooled for 10 minutes to 5 hours, then immersed in an extraction solvent at 50 to 80 캜 for 10 minutes to 5 hours, . At this time, the extraction solvent used for the cold beating and the extraction solvent used for warming are preferably the same extraction solvent.

Each of the extraction solvents for extracting the extract may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. As the above-mentioned extraction solvent, although not particularly limited, an extract extracted with ethanol or water is preferably used for the improvement, prevention or treatment of diabetes, that is, insulin-dependent diabetes (type 1) or non-insulin dependent diabetes mellitus (type 2).

In the present invention, the term "diabetes" means a disease that occurs when insulin secretion is insufficient or insulin action and function are not sufficiently achieved. In case of this disease, excessive decomposition of glycogen, protein, Resulting in diabetes and ketoneuria, resulting in abnormal blood and electrolyte metabolism, and concomitant conditions such as circulatory disturbances and renal disorders, as well as blood concentration due to electrolyte loss. Insulin is secreted in the beta cells of the islets of the islets in the pancreas, secreted when the glucose level in the blood is increased, and secreted is suppressed when the blood glucose level is increased. The disease is divided into insulin - dependent diabetes mellitus (type I) and non - insulin dependent diabetes mellitus (type II). Diagnosis of diabetes is usually made by measuring blood glucose levels, which vary according to standards. In humans, diabetes is usually diagnosed when the blood glucose level is more than 200 mg / dl, and fasting is more than 140 mg / dl. Thus, the damaged pancreatic beta cells can be repaired or the pancreas can also be treated to increase or decrease the intracellular glucose uptake, thereby treating or preventing diabetes. The inventors of the present invention have found that when the extract of the present invention is treated or the fraction thereof, the size of the damaged pancreas and the damaged pancreas increase intracellular glucose uptake, the composition can be effectively used for preventing or treating insulin-dependent diabetes mellitus Respectively.

The above-mentioned mixed solvent is not particularly limited, but an extract obtained by extracting with 20 to 95% by volume of methanol, ethanol, butanol or propanol aqueous solution, more preferably 60 to 95% by volume of ethanol aqueous solution is preferable.

The term &quot; extract &quot; used herein in reference to a sprout includes not only the crude extract obtained by treating the extraction solvent but also the extract of the extract of Gracilariopsis chorda (Holmes) Ohmi . For example, extracts of Gracilariopsis chorda (Holmes) Ohmi may be prepared in powder form by further processes such as vacuum distillation and lyophilization or spray drying.

In addition, the extract of the present invention has a meaning including a powder of a dog cocoon formulated to be able to be administered to an animal in a broad sense, such as a dog cocoon powder.

It will be appreciated by those skilled in the art that although the present invention has been tested with a twisted pair, the desired effect can be achieved in the form of a Gracilariopsis chorda (Holmes) Ohmi work piece.

As used herein, the term &quot; comprising as an active ingredient &quot; is meant to include an amount sufficient to achieve the efficacy or activity of the extract. For example, the extract is used at a concentration of 10 to 1500 μg / ml, preferably 100 to 1000 μg / ml. Since the extract of Puppet is a natural product, there is no adverse effect on human body even when it is administered in an excessive amount, so that the quantitative upper limit of the Puppet extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

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

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

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

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .

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

The pharmaceutical composition of the present invention can be prepared in unit dose form by formulating it with a pharmaceutically acceptable carrier and / or excipient or can be manufactured by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

Another aspect of the present invention relates to a pharmaceutical composition for the treatment or prevention of diabetes characterized by containing a fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

The term "fraction " of the present invention means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents. In the present invention, specifically, extracts of the above-mentioned dogs can be obtained by various methods such as a solvent fractionation method, an ultrafiltration fractionation method, and a chromatography fractionation method, but the solvent fractionation method is used in the present invention.

The Gracilariopsis chorda (Holmes) Ohmi fraction is mixed with 5 to 30-fold (w / w), preferably 8 to 20-fold (w / w) water in the concentrated canine extract, , A conventional fractionation process using a nonpolar solvent is performed to obtain a polar solvent fraction and a nonpolar solvent fraction, respectively. Examples of the polar solvent include n-butanol and water, and examples of the non-polar solvent include hexane, methylene chloride, ethyl acetate and the like. Most preferably, the Gracilariopsis chorda (Holmes) Ohmi fraction is obtained by sequentially fractionating the extract of Gracilariopsis chorda (Holmes) Ohmi with hexane, methylene chloride, ethyl acetate and n-butanol, Hexane fraction, methylene chloride fraction, or water fraction. Compared with the other fractions, the increase in pancreatic ductility was markedly greater than in the untreated control, and the size of the reduced pancreatic duct was significantly increased , And increased intracellular glucose uptake was confirmed.

The term "fraction" as used herein in reference to a sprout includes not only fractions obtained by treating extraction solvents, but also fractions of fractions of Gracilariopsis chorda (Holmes) Ohmi . For example, the Gracilariopsis chorda (Holmes) Ohmi fraction may be prepared in powder form by further processes such as vacuum distillation and lyophilization or spray drying.

In addition, the canine fraction of the present invention has a meaning including a powder of a canine, which is formulated so that the canine can be administered to an animal in a broad sense, for example, a powder of canine corn.

It will be appreciated by those skilled in the art that although the present invention has been tested with a twisted pair, the desired effect can be achieved in the form of a Gracilariopsis chorda (Holmes) Ohmi work piece.

By the term &quot; comprising as an active ingredient &quot;, as used herein, is meant to include an amount sufficient to achieve the efficacy or activity of the cuticle fraction. For example, the open-eye cut fraction is used at a concentration of 10 to 1500 μg / ml, preferably 100 to 1000 μg / ml. Since the fraction of Pythagorea sp. Is a natural product, there is no adverse effect on the human body even when administered in an excessive amount. Therefore, the quantitative upper limit of the Pythagoreaceae fraction contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

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

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

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

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .

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

The pharmaceutical composition of the present invention can be prepared in unit dose form by formulating it with a pharmaceutically acceptable carrier and / or excipient or can be manufactured by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

In addition, the present invention provides a food composition for improving or preventing diabetes containing an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolates, pizza, ram noodles, other noodles, gums, ice cream, .

The food composition of the present invention may contain, as an active ingredient, an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi , as well as components which are conventionally added in the manufacture of a food product, for example, proteins, carbohydrates, fats, Nutrients, flavoring agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and a beverage, it may contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, and various kinds of extracts or fractions of Gracilariopsis chorda (Holmes) Plant extracts, and the like.

The present invention provides a health functional food comprising a food composition for improving, preventing or treating diabetes comprising the extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient. Health functional foods are foods prepared by adding extracts or fractions of Gracilariopsis chorda (Holmes) Ohmi to food materials such as beverages, tea, spices, gum and confectionery, or by encapsulation, powdering, suspension and the like When taken, it means bringing about a certain effect on health, but unlike general medicine, there is an advantage that there is no side effect that can occur when a medicine is taken for a long time by using food as a raw material. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely. The amount of the extract or fraction of Gracilariopsis chorda (Holmes) Ohmi in such a health functional food can not be uniformly determined depending on the type of the health functional food to be targeted, but it is preferably within a range that does not impair the original taste of the food And is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight based on the target food. In the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added usually in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

The present invention also provides a method of improving, preventing or treating diabetes comprising administering to a mammal an effective amount of an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi .

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

As used herein, the term "effective amount" refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal, or human, as contemplated by a researcher, veterinarian, physician or other clinician, &Lt; / RTI &gt; inducing a reduction of the symptoms of the disease or disorder. The effective amount and the administration frequency for the active ingredient of the present invention can be changed according to the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the prevention, treatment, or improvement method of the present invention, a dose of 0.001 g / kg to 10 g / kg of a Gracilariopsis chorda (Holmes) Ohmi extract or fraction is administered once to several times a day, Lt; / RTI &gt;

The composition comprising the extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient in the treatment method of the present invention can be administered orally or rectally, intravenously, intraarterally, intraperitoneally, intramuscularly, intrasternally, percutaneously, Can be administered in a conventional manner via intraocular or intradermal routes.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example 1. Preparation of ethanol extract of canola.

The freeze-dried canine corn (2.84 kg) was sieved, and the whole was frozen twice with ethanol (17 L) at room temperature. Then, it was warmed once at 60 ° C in a water bath and concentrated to obtain an ethanol extract of canola (hereinafter referred to as "&Lt; / RTI &gt;

Example 2: Preparation of hot water extract of corn borer.

The lyophilized canola (100 g) was sieved and extracted twice with 3 L of hot water (50 L) for 3 hours. The extracted solution was filtered to collect only the extract, which was then concentrated to give a sea tangle extract Called hot-water extract).

Examples 3 to 7. Preparation of cornflakes fractions.

The freeze-dried canine corn (2.84 kg) was sieved, and the whole was frozen twice with ethanol (17 L) at room temperature. Then, it was warmed once at 60 ° C in a water bath and concentrated to obtain an ethanol extract of canola (hereinafter referred to as " To obtain 166.58 g.

161.58 g of the above extract was suspended in 1.6 L of distilled water and fractionated three times with hexane, methylene chloride, ethyl acetate and n-butanol, respectively, according to a conventional method to obtain 27.64 g of a hexane fraction (Example 3) and a methylene chloride fraction 4), 0.15 g of the ethyl acetate fraction (Example 5), 4.80 g of the n-butanol fraction (Example 6) and 126.18 g of the water fraction (Example 7). Its specific procedure is shown in Fig.

Experimental Example 1. Measurement of zebrafish pancreatic islet change

Alloxan monohydrate, sea salt, and insulin were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

1) Zebrafish breeding

The adult zebrafish was maintained in a zebra fish system S type (W 1500 X D 400 X H 2050) (Genomic Design Bioengineering Co., Korea), and was repeatedly lighted for 14 hours at 28.5 ° C and dark for 10 hours. To obtain the zebra fish larvae, two pairs of adult zebra fish were placed in the scattering box overnight. The next day, zebrafish began laying eggs while giving a half-hour light. Then, for the cultivation of zebrafish fish, zebrafish fish were collected 3 hours after fertilization, and they were collected in an incubator at 28.5 ° C in a light and dark photoperiod of 14:10 and stored in a solution containing 0.03% sea salt Respectively.

2) Zebrafish pancreatic islet change experiment

The zebrafish larvae were divided into groups treated with the zebrafish larvae, the aloe vera or the insulin-treated untreated control, the canine extracts and the fractions, and the staining with 2-NBDG was followed by fluorescence microscopy to measure changes in the pancreatic islets . Specific experimental conditions were described for each experimental example.

3) Statistical analysis

Statistical analysis was performed using Graphpad Prism (version 5). The data are expressed as averages of the standard error of mean (SEM). Efficacy was assessed using the Tukey HSD test and one-way ANOVA. The likelihood level for statistical validity was P <0.05.

Experimental Example 2. Analysis of antidiabetic effect by short-term administration of an extract of dog leucocyte in an allose-induced diabetic zebrafish

The zebrafish larvae were divided into the zebrafish larval normal group, the unoxidized control group induced by alloxan, and the group treated with the ethanol extract of the dog corn (Example 1) and the hot water extract of the dog corn (Example 2).

1) Experimental method

Five days post-fertilization zebrafish larvae were placed in 96 wells and stained with 25 μM 2-NBDG for 12 hours. After exposure to 100 μM alloys for 15 min, the cells were replaced with 0.03% sea salt solution and exposed for up to 5 h. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation, and 25 μM 2-NBDG was re-stained for 1 h before fluorescence microscopy. After the first microscope photographing, 10 μg / mL of each of the extracts of Example 1 and Example 2 was exposed for 1 hour, and the group treated with the ethanol extract of Example 1 (Example 1) and the group treated with hot water extract of canine starch (Example 2) Was prepared. To observe changes in pancreatic islets in each of the above groups, a second microscopic photograph was taken and 25 μM 2-NBDG was re-stained for 1 hour before fluorescence microscopy as in the first microscopic photograph. Images obtained through fluorescence microscopy were analyzed for pancreatic islet size in zebrafish using Focus Lite software.

2) Results

In order to confirm the anti-diabetic effect when the short-term administration of the ethanol extract of the dog fish prepared in Example 1 and the hot water extract of the dog fish prepared in Example 2 in the alooxan-induced diabetic zebrafish model, FIG. 3 is a graph showing the measurement of pancreatic islet fluorescence of each group. FIG.

As shown in FIG. 2 and FIG. 3, zebrafish having impaired pancreatic islets was prepared by using alooxan for the analysis of antidiabetic effect. When aloxanic acid was used, the pancreas of zebrafish was 200 times As shown in Fig.

Among the canine extracts prepared from Example 1 and Example 2, the extract of Example 1 was found to have a strong antidiabetic activity at a concentration of 10 μg / ml. In the group treated with the ethanol extract of cane extract of Example 1, it was confirmed that the pancreatic islet was more than 7 times more than the normal group, even though the effect of pancreatic islet reduction induced by alloxan was overcome. The hot-water extract of canine corn in Example 2 also tended to overcome the decrease in pancreatic islets, but no significant difference was observed.

Experimental Example 3. Anti-diabetic effect analysis by short-term administration of the macrocyclic fraction in the aloic acid-induced diabetic zebrafish

The zebrafish larvae were divided into the zebrafish larval normal group, the aloic acid-derived untreated control group, and the group treated with each of the dog cauliflower fractions prepared in Examples 3 to 7, respectively.

1) Experimental method

Five days post-fertilization zebrafish larvae were placed in 96 wells and stained with 25 μM 2-NBDG for 12 hours. After exposure to 100 μM alloys for 15 min, the cells were replaced with 0.03% sea salt solution and exposed for up to 5 h. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation, and 25 μM 2-NBDG was re-stained for 1 h before fluorescence microscopy. After the primary microscopic photographing, the dog cuticles prepared from Examples 3 to 7 were exposed to each 10 占 퐂 / ml for 1 hour to prepare groups treated with respective canine fractions (Examples 3 to 7). To observe changes in pancreatic islets in each of the above groups, a second microscopic photograph was taken and 25 μM 2-NBDG was re-stained for 1 hour before fluorescence microscopy as in the first microscopic photograph. Images obtained through fluorescence microscopy were analyzed for pancreatic islet size in zebrafish using Focus Lite software.

2) Results

Diabetic zebra fish model induced by alloxan was used for confirming antidiabetic effect when short-term administration of the dog cauliflower fractions prepared in Examples 3 to 7 was performed. Fig. 4 is a graph showing changes in size of pancreatic islets in each group And FIG. 5 is a graph showing measurement of fluorescence intensity of pancreatic islets in each group.

The cuticle cuts of Examples 3 to 7 showed a tendency to overcome the decrease of pancreatic islets, and the cuttlefish hexane fraction prepared from Example 3 showed significant antidiabetic effect. The group treated with the open-cut hexane fraction of Example 3 showed that the pancreatic islets were more increased than the normal group, even though they overcome the effects of pancreatic islet induced by alloxan.

Experimental Example 4. Antidiabetic effect by long-term administration of an extract of dog cauliflower in an allose-induced diabetic zebrafish

Zebrafish larvae were divided into groups treated with zebrafish larvae normal, aloic acid or insulin-treated untreated control group, and the extracts of dogs prepared from Example 1 and Example 2 were treated with 1 μg / ml and 10 μg / ml, respectively Respectively.

1) Experimental method

Five days post-fertilization zebrafish larvae were placed in 96 wells and exposed to 100 μM alloys for 6 hours. Then, each group was treated with the extract of corn borer (Ex. 1, 2) (1, 10 占 퐂 / ml) by exposing the corn borer extract to a concentration of 1, 10 占 퐂 / ml for 12 hours. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation and 40 μM 2-NBDG was stained for 30 min before fluorescence microscopy and exposed to 0.03% saline solution for 20 min. Images obtained through fluorescence microscopy were analyzed for the intensity of the pancreatic islets of the zebrafish and the intensity of the fluorescence intensity of the pancreatic islets using Focus Lite and Image J software.

2) Results

The diabetic zebrafish model induced by alloxan was used to confirm the anti-diabetic effect when the dog extract prepared from Example 1 and Example 2 was administered for a long time. Fig. 6 shows the change of pancreatic islet size in each group And FIG. 7 is a graph showing the measurement of the fluorescence intensity of pancreatic islets in each group.

As a result, the pancreatic islet size of the group treated with 10 ㎍ / ㎖ of the extract of Example 1 was significantly higher than that of the untreated control group, and the fluorescence of the pancreatic islets was also significantly increased.

In addition, the fluorescence intensity of the pancreatic islets was significantly increased even in the group treated with 10 ㎍ / ㎖ of the hot-water extract of canine corn extract of Example 2, but it was found to be relatively weaker than that of the canine corn ethanol extract of Example 1 .

Experimental Example 5. Analysis of antidiabetic effect by long-term administration of the macrocyclic fraction in the aloic acid-induced diabetic zebrafish

The zebrafish larvae were divided into a group treated with zebrafish normal group, an untreated control group induced with alloxan, and a group treated with 1 ㎍ / ㎖ and 10 / / ㎖ of the dog cauliflower fractions prepared in Examples 3 to 7, respectively Respectively.

1) Experimental method

Five days post-fertilization zebrafish larvae were placed in 96 wells and exposed to 100 μM alloys for 6 hours. After that, the cuticle fractions prepared from Examples 3 to 7 were treated with each of the cuticle fractions (Examples 3 to 7) (1, 10 占 퐂 / ml) for 12 hours at a concentration of 1, 10 占 퐂 / Group. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation and 40 μM 2-NBDG was stained for 30 min before fluorescence microscopy and exposed to 0.03% saline solution for 20 min. Images obtained through fluorescence microscopy were analyzed for the intensity of the pancreatic islets of the zebrafish and the intensity of the fluorescence intensity of the pancreatic islets using Focus Lite and Image J software.

2) Results

In order to confirm the antidiabetic effect when the dog extract (1 μg / ml, 10 μg / ml) prepared in Examples 3 to 7 was administered for a long time in the dirogenic zebrafish model induced by alloxan, FIG. 9 is a graph showing the measurement of pancreatic islet fluorescence of each group. FIG.

As shown in Figs. 8 and 9, the anti-diabetic activity test results of the dog's cuticle solvent fractions obtained in Examples 3 to 7 showed that the pancreatic islet size was significantly higher at a concentration of 1 / / ml of the open mouth methylene chloride fraction (Example 4) (Example 4), the methylene chloride fraction (Example 4), the water fraction (Example 7), and the methylene chloride fraction (Example 4), respectively, and the fluorescence intensity of the pancreatic islets was increased to 1 μg / (10 μg / ml) of the methylene chloride fraction (Example 4) showed an effect similar to that of glymefamide, which is a therapeutic agent for diabetes mellitus.

Experimental Example 6: Antidiabetic effect of the extract of dogs on the insulin-induced diabetic zebrafish.

Zebrafish larvae were treated with 1 ㎍ / ㎖ and 10 ㎍ / ㎖ of zebrafish larvae normal group, insulin-treated untreated control group, dog canine ethanol extract (Example 1) and canine hot water extract (Example 2) Group.

1) Experimental method

Three days post-fertilization zebrafish larvae were placed in 6 wells and exposed to 10 μM insulin for 48 hours. Then, the extracts prepared in Examples 1 and 2 were exposed to a concentration of 1, 10 / / ml for 48 hours. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation and stained with 40 μM 2-NBDG for 30 min before fluorescence microscopy. Images obtained through fluorescence microscopy were analyzed for pancreatic islet size of zebrafish using Focus Lite software.

2) Results

In order to confirm the antidiabetic effect of the extract of dogs (1 占 퐂 / ml, 10 占 퐂 / ml) prepared in Examples 1 and 2 in a diabetic zebra fish model induced by excessive insulin, Fig. FIG. 11 is a graph showing fluorescence intensity of pancreatic islets in each group. FIG.

As shown in Figs. 10 and 11, it was confirmed that the pancreatic islet size and the fluorescence intensity of the pancreatic islet significantly increased in the group treated with 1 占 퐂 / ml of the ethanol extract of cane extract of Example 1.

Experimental Example 7: Anti-diabetic effect of the extracorporeal fraction in diabetic zebrafish induced by excessive insulin.

The zebrafish larvae were analyzed by dividing into zebrafish larval normal group, insulin-induced untreated control group, and the group of dogs of Examples 3 to 7 treated with 1 占 퐂 / ml and 10 占 퐂 / ml, respectively.

1) Experimental method

Three days post-fertilization zebrafish larvae were placed in 6 wells and exposed to 10 μM insulin for 48 hours. Then, the cuticle fractions prepared in Examples 3 to 7 were exposed for 48 hours at a concentration of 1, 10 μg / ml. Fluorescence microscopy (Olympus 1 × 70, Olympus, Japan) was used for direct pancreatic islet observation and stained with 40 μM 2-NBDG for 30 min before fluorescence microscopy. Images obtained through fluorescence microscopy were analyzed for pancreatic islet size of zebrafish using Focus Lite software.

2) Results

In order to confirm the antidiabetic effect of the dog cauliflower fractions (1 占 퐂 / ml, 10 占 퐂 / ml) prepared from Examples 3 to 7 in a diabetic zebra fish model caused by excessive insulin, Fig. FIG. 13 is a graph showing the fluorescence intensity of the pancreatic islets measured in each group. FIG. ^{(## p <0.01, ### p} <0.001; control group and bigyodoem), (* p <0.05, ** p <0.01, *** p <0.001; bigyodoem and untreated control group)

As shown in Figs. 12 and 13, the results of the antidiabetic activity tests of the dog cage fractions obtained in Examples 3 to 7, the size of pancreatic islets in the dog cage hexane fraction of Example 3 or the dog cage methylene chloride fraction of Example 4 And fluorescence of pancreatic islets tended to increase.

As a result, it can be seen that the extracts or fractions of Examples 1 to 6 according to the present invention can be stably used as a pharmaceutical composition, a cosmetic composition or a food composition, since the raw materials are based on natural products .

Experimental Example 8. Confirmation of Side Effects of Extracts

Morphological changes and other toxic responses were not observed when zebrafish embryos were treated with the same concentrations used in Example 1, 2 corn extracts and the cornflower fractions of Examples 3 through 7 in the antidiabetic efficacy test.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

Claims (9)

A pharmaceutical composition for the treatment or prevention of diabetes, which comprises an extract of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient. The method according to claim 1,
Wherein the extract of Gracilariopsis chorda (Holmes) Ohmi) is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. 3. The method of claim 2,
Wherein the mixed solvent is an aqueous solution of methanol, ethanol, butanol or propanol in an amount of 20 to 95% by volume. The method according to claim 1,
Wherein said diabetes is type 2 diabetes caused by decreased type 1 diabetes caused by reduction of the degree of pancreas induced by alloxan or by reduction of pancreas induced by excessive insulin. . A pharmaceutical composition for the treatment or prevention of diabetes characterized by containing a fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient. 6. The method of claim 5,
The Gracilariopsis chorda (Holmes) Ohmi fraction is a hexane fraction obtained by sequential fractionation of Gracilariopsis chorda (Holmes) Ohmi extract with hexane, methylene chloride, ethyl acetate, n-butanol, methylene chloride and And water fraction. The pharmaceutical composition for treating or preventing diabetes according to claim 1, 6. The method of claim 5,
Wherein said diabetes is type 2 diabetes caused by decreased type 1 diabetes caused by reduction of the degree of pancreas induced by alloxan or by reduction of pancreas induced by excessive insulin. . A composition for improving or preventing diabetes, which comprises an extract or fraction of Gracilariopsis chorda (Holmes) Ohmi as an active ingredient. 9. The method of claim 8,
Wherein said diabetes is type 2 diabetes caused by reduction of pancreas induced by alloxan induced type 1 diabetes or excess insulin induced decrease in pancreas level .

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