KR20170039457A - Composition for preventing or treating diabetes comprising a flavone-based compound from Kaempferia parviflora - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of diabetes, which comprises a black ginger-derived flavonoid compound. More particularly, the present invention relates to a pharmaceutical composition for preventing and treating diabetes mellitus comprising 6-hydroxy-7,4'-dimethoxyflavone or a pharmaceutically acceptable salt thereof. The present invention relates to a pharmaceutical composition for preventing and treating diabetes, which can reduce blood sugar, increase insulin secretion amount, and has a low toxicity and less side effects.

Description

[0001] The present invention relates to a pharmaceutical composition for prevention and treatment of diabetes, comprising a flavonoid compound derived from black ginger,

The present invention relates to a pharmaceutical composition for the prevention and treatment of diabetes.

The black ginger (kaempferia parviflora Wall. Ex Baker) is a root-stemmed plant of the native plant of Thailand. It shows purple or black inside when cut immediately after collection. The black ginger has been known to help maintain mental and physical stability and stable blood pressure, to treat dysentery and to treat abdominal pain.

However, it has not yet been reported that black ginger has an effect on diabetes. The major part of the study of black ginger is the methoxy flavone, which is a representative component of the gangrene.

As a major study on the active ingredient, it has been reported that Lupeol is effective in the treatment of inflammation in rats. Lupeol has been reported to inhibit cancer by suppressing NF-kB, P13K, and AKt, which are the skin cancer mechanisms, and animal experiments that protect the liver by the effects of antioxidants. There is also a study on the components of Oxyresveratrol, which studies the effects of antioxidants to inhibit oxidation, but it has been found that 6-hydroxy-7,4'-dimethoxyflavone isolated from black ginger of the present invention or its pharmaceutically acceptable Is different from a composition for treating diabetes containing a salt as an active ingredient.

The present invention relates to a composition for treating diabetes mellitus containing 6-hydroxy-7,4'-dimethoxyflavone compound, a pharmaceutically acceptable salt thereof or a black ginger extract containing the compound as an active ingredient. More particularly, 6-hydroxy-7,4'-dimethoxyflavone isolated from black ginger, which has excellent diabetic effect, may be useful as a pharmaceutical composition for the prevention and treatment of diabetes or as a health food for the prevention of diabetes.

In the present invention, 6-hydroxy-7,4'-dimethoxyflavone among the components extracted from black ginger was tested for antidiabetic effect in streptozotocin (STZ) induced diabetic rats. As a result, STZ-induced diabetic rats received 50 mg / / Kg and 150 mg / Kg, respectively. After 4 weeks of blood glucose measurement, insulin, Triglyceride and C-peptide were obtained from the serum obtained from the dissection after the measurement. The recommended triglyceride levels were not significantly different, and the insulin, C-peptide, and blood glucose levels were directly measured. In the above results, the 6-hydroxy-7,4'-dimethoxyflavone component extracted from black ginger appears to be effective for the treatment of diabetes by increasing the insulin concentration and decreasing the blood glucose in the STZ-induced diabetic rats. It has succeeded in isolating and purifying 6-hydroxy-7,4'-dimethoxyflavone, a novel low-toxicity diuretic substance of new active ingredient.

Published Patent Application No. 2015-0040618 (Apr. 15, 2015)

It is an object of the present invention to provide a pharmaceutical composition for the prevention and treatment of diabetes.

It is another object of the present invention to provide a food composition for improving and preventing diabetes.

It is another object of the present invention to provide a method for extracting 6-hydroxy-7,4'-dimethoxyflavone from black ginger.

1. A composition for the prevention and treatment of diabetes comprising 6-hydroxy-7,4'-dimethoxyflavone or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition for preventing or treating diabetes according to 1 above, wherein the 6-hydroxy-7,4'-dimethoxyflavone is obtained from black ginger extract.

3. The pharmaceutical composition for preventing or treating diabetes according to 2 above, wherein the black ginger extract is extracted with an organic solvent.

4. The pharmaceutical composition for prevention or treatment of diabetes according to 3 above, wherein the black ginger extract is obtained from the ethyl acetate fraction by further performing an ethyl acetate fraction after the organic solvent extraction.

5. The pharmaceutical composition for preventing or treating diabetes according to item 3 above, wherein the organic solvent is a C1 to C4 alcohol.

6. The pharmaceutical composition for preventing or treating diabetes according to item 5 above, wherein the C1 to C4 alcohol is methanol.

7. The pharmaceutical composition for preventing or treating diabetes according to 1 above, wherein a single dose of the composition is such that 6-hydroxy-7,4'-dimethoxyflavone is contained in an amount of 50 to 200 mg per 1 kg of body weight of the subject to be administered.

8. A food composition for improving or preventing diabetes comprising 6-hydroxy-7,4'-dimethoxyflavone.

9. Extracting black ginger with methanol; Suspending the methanol extract in water; And extracting 6-hydroxy-7,4'-dimethoxyflavone from black ginger comprising adding ethyl acetate to the suspension and obtaining an ethyl acetate fraction.

The composition of the present invention can be used for prevention and treatment of diabetes.

The composition of the present invention can reduce blood sugar in diabetic patients.

The composition of the present invention can increase insulin secretion amount in a diabetic patient.

The compositions of the present invention are less toxic and less of a side effect problem.

According to the extraction method of the present invention, 6-hydroxy-7,4'-dimethoxyflavone can be extracted from black ginger.

FIG. 1 shows the result of 6-hydroxy-7,4'-dimethoxyflavone as a result of 1 H-NMR analysis.
FIG. 2 is a graph comparing changes in blood glucose level when 6-hydroxy-7,4'-dimethoxyflavone is treated with streptozotocin (STZ) in induced diabetic rats.
Figure 3 compares C-peptide changes in 6-hydroxy-7,4'-dimethoxyflavone treated with streptozotocin (STZ) induced diabetic rats.
Figure 4 compares insulin changes in 6-hydroxy-7,4'-dimethoxyflavone treated with streptozotocin (STZ) induced diabetic rats.
FIG. 5 compares the changes of 6-hydroxy-7,4'-dimethoxyflavone with Triglyceride in streptozotocin (STZ) -induced diabetic rats.

The present invention relates to a composition for preventing and treating diabetes mellitus containing a black ginger-derived flavonoid compound, and more particularly, to a composition for preventing and treating diabetes mellitus comprising 6-hydroxy-7,4'-dimethoxyflavone or a pharmaceutically acceptable salt thereof, Which can increase the insulin secretion amount, and which has a low toxicity and thus less of side effects.

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

The present invention provides a composition for treating diabetes comprising 6-hydroxy-7,4'-dimethoxyflavone compound represented by the following formula (1), a pharmaceutically acceptable salt thereof or a black ginger extract containing the compound as an active ingredient .

[Chemical Formula 1]

In the composition for treating diabetes mellitus of the present invention, the 6-hydroxy-7,4'-dimethoxyflavone compound represented by the formula (1) can be isolated from black ginger. In the separation method, the black ginger is extracted with an organic solvent, and the extracted extract is suspended in water, followed by treatment with ethyl acetate to obtain an ethyl acetate fraction. From the obtained ethyl acetate fraction, the compound of Formula 1 can be obtained. But is not limited thereto.

The organic solvent is preferably an alcohol such as an alcohol of C1 to C4, and the alcohol is more preferably methanol, but is not limited thereto.

The present invention also relates to the use of the 6-hydroxy-7,4'-dimethoxyflavone compound represented by the above formula (1), a pharmaceutically acceptable salt thereof or a black ginger extract containing the above compound as an active ingredient for the prevention Or a pharmaceutical composition for therapeutic use. In the pharmaceutical composition for preventing or treating diabetes mellitus according to the present invention, the 6-hydroxy-7,4'-dimethoxyflavone has a function of treating diabetes mellitus, and the disease can be prevented or treated by this function.

"Pharmaceutically acceptable salt" as used herein refers to any salt of the compound of the present invention that does not exhibit toxicity or is unsuitable for pharmaceutical use while maintaining the biological properties of the compounds of the present invention. Such salts may be derived from, and include, various organic and inorganic counterions known in the art.

Such salts include (1) organic or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfamic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, But are not limited to, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) Butanedicarboxylic acid, 4-chlorobenzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, benzenesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butyl acetic acid, lauryl sulfuric acid, Gluconic acid, benzoic acid, glutamic acid, hydroxynaphthoic acid, Salicylic acid, stearic acid, cyclohexylsulfamic acid, quinic acid, muconic acid and the like; Or (2) the acidic proton present in the parent compound is selected from the group consisting of (a) metal ions (e.g., alkali metal ions, alkaline earth metal ions or aluminum ions), alkali metal or alkaline earth metal hydroxides (e.g., sodium, potassium, calcium, magnesium (B) an organic base such as an aliphatic, alicyclic or aromatic organic amine such as ammonia, methylamine, dimethylamine, diethylamine, picoline , Ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N, N'-dibenzylethylene-diamine, chloropropane, diethanolamine, procaine, N- Amine, N-methylglucamine piperazine, tris (hydroxymethyl) -aminomethane, salts formed when coordinating with tetramethylammonium hydroxide, and the like. Examples of salts include sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium salts. When the compound contains a basic functional group, salts with non-toxic organic acids or inorganic acids such as hydrohalides (for example, Chloride or hydrobromide), sulfate, phosphate, sulfamate, nitrate, acetate, trifluoroacetate, trichloroacetate, propionate, hexanoate, cyclopentylpropionate, glycolate, glutarate, (4-hydroxybenzoyl) fumarate, tartarate, tartrate, malate, succinate, ascorbate, malate, maleate, fumarate, tartrate, citrate, benzoate, Citrate, benzoate, 3- (4-hydroxybenzoyl) benzoate, picrate, cinnamate, , Phthalate, laurate, methanesulfonate mesylate), ethanesulfonate, 1,2-ethane-disulfonate, 2-hydroxyethanesulfonate, benzenesulfonate (besylate), 4-chlorobenzenesulfonate, 2-naphthalenesulfonate, 4-toluenesulfonate, camphorate, camphorsulfonate, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylate, glucoheptonate, 3- Propionate, trimethylacetate, tert-butyl acetate, lauryl sulfate, gluconate, benzoate, glutamate, hydroxynaphthoate, salicylate, stearate, cyclohexyl sulfamate, quinate, .

Also, in the present invention, the compounds of the present invention may have one or more asymmetric centers, and thus such compounds may be prepared from individual (R) -enantiomers or (S) -enantiomers, or mixtures thereof. Unless otherwise indicated, for example, the description or designation of certain compounds in the specification and claims by stereochemical nomenclature at any position in the formulas may be applied to individual enantiomers and their racemic mixtures or other mixtures, . Stereochemical crystallization methods and stereoisomeric separation methods are well known in the art. The present invention also provides stereoisomers of the compounds shown in this invention upon treatment with a base.

The term "therapeutic agent (s) or therapeutic composition" as used herein refers to any agent (s) that can be used to treat or prevent a disorder or one or more symptoms thereof. In certain embodiments, the term "therapeutic agent" refers to a compound of the invention. In another particular embodiment, the term "therapeutic agent" does not refer to a compound of the invention. In one embodiment, the therapeutic agent is an agent that is known to be useful for treating or preventing a disorder or one or more of its symptoms, or that has been or has been used for that purpose.

"Therapeutically effective amount" means an amount of a compound, complex or composition sufficient to be effective in treating the disease when administered to a subject for treatment of the disease. The "therapeutically effective amount" may vary depending, inter alia, on the compound, the disease and its severity, and the age, body weight, In one embodiment, "treating" any disease or disorder refers to ameliorating a disease or disorder present in the subject. In another embodiment, "treatment" refers to ameliorating one or more body parameters, and the subject may not be aware of it. In another embodiment, "treatment" is intended to include the treatment of a disease or disorder with a physiological control (e. G., Stabilization of a physical parameter) It represents both.

In another embodiment, "treatment" refers to delaying the onset of the disease or disorder. The term "prophylactic agent (s) " as used herein refers to any agent (s) that can be used to prevent the disorder or one or more symptoms thereof. In certain embodiments, the term "prodrug" refers to a compound of the invention.

In another specific embodiment, the term "preventive composition" does not refer to a compound of the present invention. For example, the composition for prevention is an agent which has been known to be useful for preventing or delaying the onset, development, progression and / or deepening of a disorder, or which has been or has been used for this purpose.

The term "prevention ", as used herein, refers to administration of one or more symptoms of a disorder in a subject by administering a combination of therapies (e.g., prophylactic or therapeutic agents) or combinations of therapies (e.g., ≪ RTI ID = 0.0 > recurrence, < / RTI > A "prophylactically effective amount" refers to an amount sufficient to prevent the development, recurrence, or onset of one or more symptoms associated with a disorder, or to increase or improve the prophylactic or therapeutic effect (s) of another therapeutic agent (For example, prophylactic agent). The compounds used in the method of the present invention are preferably used in combination with one or more compounds of formula I, where appropriate in salt form, either alone or in combination with one or more suitable pharmaceutical acceptable carriers such as diluents or adjuvants , ≪ / RTI > or another nerve cell protective use.

The compounds of the present invention may be used as tablets, pills, hard gelatine capsules, powders or granules as a solid composition for oral administration. In these compositions, the compounds according to the invention are mixed with one or more inert diluents or adjuvants such as sucrose, lactose or starch. These compositions may contain, in addition to diluents, materials such as, for example, lubricants such as magnesium stearate, or coatings for controlled release.

As a liquid composition for oral administration, pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing an inert diluent such as water or liquid paraffin can be used. These compositions may also contain, in addition to diluents, substances such as, for example, wetting agents, sweeteners or flavoring agents. In addition, the composition of the present invention is a pharmaceutical composition or a single unit dosage form.

The pharmaceutical compositions and single unit dosage forms of the invention comprise a prophylactically or therapeutically effective amount of one or more prophylactic or therapeutic agents (e.g., a compound of the invention, or other prophylactic or therapeutic agent), and typically one or more pharmaceutically acceptable Carrier or excipient.

In certain embodiments and contexts, the term "pharmaceutically acceptable" refers to a pharmacologically acceptable pharmacopoeia approved by a government or government regulatory agency that may be used in an animal, more particularly a bird or mammal, And the like. The term "carrier" refers to a diluent, such as a diluent, such as Freund ' s adjuvant (complete and incomplete adjuvant), excipient or vehicle, which is administered with the therapeutic. Such pharmaceutical carriers may be sterile liquids such as water, and oils, including oils or synthetic oils, of petroleum, animal or vegetable origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously.

Saline solution, and dextrose and glycerol aqueous solutions can also be used as a liquid carrier, especially a injectable solution. An example of a suitable pharmaceutical carrier is described in "Remington ' s Pharmaceutical Sciences" by E. W. Martin.

Conventional pharmaceutical compositions and dosage forms comprise one or more excipients. Suitable excipients are well known to those skilled in the art of pharmacy and non-limiting examples of suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, hull, silica gel, sodium stearate, glycerol monostearate, Talc, sodium chloride, skim milk, glycerol, propylene, glycol, water, ethanol and the like. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form will depend upon a variety of factors known in the art including the manner in which the dosage form is administered to the subject and the particular active ingredient involved in the dosage form,

Optionally, the composition or single unit dosage form may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The present invention also encompasses anhydrous pharmaceutical compositions and dosage forms comprising the active ingredient. For example, the addition of water is widely accepted in the pharmaceutical industry as a means of simulating long-term storage to determine the properties of the formulation (e.g., shelf life or stability) over time. For example, Jens T. Carstensen, Drug Stability: Principles Practice, 2d. Ed., Marcel Dekker, NY, NY, 1995, pp. 37980]. In fact, water and heat accelerate the decomposition of some compounds. Thus, the effects of water on the formulation can be very important, as water and / or moisture are typically generated during manufacture, handling, packaging, storage, transport and use of the formulation.

Anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low moisture components and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms comprising one or more active ingredients, including lactose, and a primary or secondary amine, are contemplated to be in an anhydrous form if they are expected to come into substantial contact with moisture and / or moisture during manufacture, packaging and / .

Anhydrous pharmaceutical compositions should be prepared and stored to maintain their anhydrous properties. It is therefore desirable to prevent the anhydrous composition from being exposed to water and to be packaged using known materials to be included in a suitable standard kit. Examples of suitable packaging include, but are not limited to, sealed foils, plastics, unit dose containers (e.g. vials), blister packs and strip packs.

The invention also encompasses pharmaceutical compositions and dosage forms comprising one or more compounds that reduce the rate of degradation of the active ingredient. Such compounds (referred to as "stabilizers" in the present invention) include, but are not limited to, antioxidants (e.g., ascorbic acid), pH buffers or salt buffers.

The pharmaceutical compositions and single unit dosage forms may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations and the like. Formulations for oral administration may include standard carriers such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like.

Such compositions and dosage forms will contain a prophylactically or therapeutically effective amount of a prophylactic or therapeutic agent, preferably in purified form, in a suitable amount and in a form suitable for administration to a subject.

The formulation should be appropriate for the mode of administration. A typical dosage form of the invention comprises a compound of the invention, or a pharmaceutically acceptable salt, solvate or hydrate thereof, in an amount ranging from about 0.1 mg to about 1000 mg / day, Administered in a single dose or, preferably, with a meal for one day. Certain dosage forms of the invention comprise a compound of Formula 1 at about 0.1, 0.2, 0.3, 0.4, 0.5, 1.0, 2.0, 2.5, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 100, 200, 250, Respectively.

Suitable pharmaceutical compositions of the present invention for oral administration may be in separate dosage forms such as tablets (e.g., chewing tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain a predetermined amount of the active ingredient and may be prepared by the preparation methods well known to those skilled in the art. Generally, see Remington ' s Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton PA (1990).

In certain embodiments, the oral dosage form is a solid and is prepared under anhydrous conditions using anhydrous ingredients as described in detail in the paragraph above. Examples of excipients that may be used in the oral dosage forms of the present invention include, but are not limited to, binders, fillers, disintegrants, and lubricants. Suitable binders for use in pharmaceutical compositions and dosage forms include corn starch, potato starch or other starch, gelatin, natural and synthetic gums such as acacia gum, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, Cellulose and derivatives thereof (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose (e.g., No. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

The present invention also provides a method for preventing or ameliorating diabetes mellitus comprising 6-hydroxy-7,4'-dimethoxyflavone compound of the above formula 1, its pharmaceutically acceptable salt or a black ginger extract containing the above compound as an active ingredient Provide health food. Examples of excipients that may be used in the oral dosage forms of the present invention include, but are not limited to, binders, fillers, disintegrants, and lubricants.

Suitable binders for use in pharmaceutical compositions and dosage forms include corn starch, potato starch or other starch, gelatin, natural and synthetic gums such as acacia gum, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, Cellulose and derivatives thereof (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropylmethylcellulose (e.g., No. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

The present invention also provides a method for preventing or ameliorating diabetes mellitus comprising 6-hydroxy-7,4'-dimethoxyflavone compound of the above formula 1, its pharmaceutically acceptable salt or a black ginger extract containing the above compound as an active ingredient Provide health food. Examples of foods to which the compound can be added include various foods, beverages, gums, tea, vitamin complexes, and health functional foods.

The amount of the compound in the food or beverage may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml. The health functional beverage composition of the present invention is not particularly limited to the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tautatin, stevia extract, for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above . The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. In addition to the above, the compound of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like.

In addition, the compounds of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. The present invention also relates to a process for the production of 6-hydroxy-7,4 ', 6'-dihydroxy-7,4'-dicarboxylic acid from the ethyl acetate fraction by extracting black ginger with an organic solvent, suspending the extracted ginger in water, treating with ethyl acetate to obtain an ethyl acetate fraction, hydroxy-7,4'-dimethoxyflavone compound represented by the formula (1), which comprises separating the compound of formula (1). The organic solvent is preferably an alcohol such as an alcohol of C1 to C4, and the alcohol is more preferably methanol, but is not limited thereto.

Hereinafter, the present invention will be described in more detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  One: Black ginger  Separation of the active component of the extracted fraction

In order to investigate the substance identification of the ethyl acetate fraction of black ginger exhibiting the effect of diabetes treatment in the present invention, black ginger was added to a concentrated flask containing 600 g of the sample at room temperature (20 to 25 degrees Celsius) . After confirming that the frozen sample is in a state of a little liquid, 300ml of the second distilled water is firstly melted and then transferred to the 2L fraction funnel. Then, 300ml of the second distilled water and the fraction 300 ml of chloroform, the solvent to be used for the first time, was added to dissolve and most of it was dissolved and transferred to a fractionation funnel in a slightly diluted liquid state. After transferring, the remaining distilled water and 300 ml of chloroform were added to the fractionation funnel, After all was recovered, all the remaining fractions were put into the fraction funnel, and the solution in the fraction funnel Secondary distilled water was made so even example by filling the remaining portion in a 2L 2L of total volume to 900ml and 600ml chloroform chloroform 500ml.

Since the chloroform is more dense than water due to the difference in density, the solution is separated by density. The solution is recovered through the coke beneath the fractionation funnel in order to recover only the chloroform solution. In the process, only the lower part of the boundary was recovered.

The process was repeated 2-3 times, and at the end, it was slowed down to the boundary point to receive the unmixed part. After the fraction of chloroform layer was finished, the water layer and ethyl acetate were added, and the mixture was shaken with a fraction funnel. The fraction of solvent in the fractionation funnel was added to the fractionation funnel with a higher amount of ethyl acetate than the amount of water layer.

Unlike the fractionation of the chloroform layer, the density of the ethyl acetate is lower than the density of the water, and it is located on the top. First, the water layer is recovered through the bottom of the coke. This time, when recovering from the Chloroform layer, Recall a little above the border. After recovery, shake the fraction funnel and shake it to recover even the water layer components on the wall. After shaking, the water layer was recovered once more and then the ethyl acetate layer was carefully withdrawn through the stomach. This process was repeated two or three times, and the end was slowed down to the boundary point to receive the unmixed part.

After finishing the ethyl acetate layer, the water layer and buthanol were added to the fractionation funnel at the same ratio or by increasing the amount of buthanol. Buthanol, like Ethyl acetate, has a lower density than water and was recovered in the same way. Unlike Ethyl acetate, Buthanol finishes in one step, finishing one time as Buthanol continues to dissolve the ingredients in water only.

The concentration of chloroform and ethylacetate was estimated to be only in the fraction of osajin sample and pomiferin sample. After completion of the fractionation of Maclura pomifera, three solvents (chloroform, Ethyl acetate, Buthanol), and 4 fractions of water were concentrated by water. The solutions were analyzed by thin layer chromatography using 6-hydroxy-7,4'-dimethoxyflavone samples. chloroform and ethylacetate were re-dissolved and concentrated into one, which was again dissolved in methanol and melted, divided into two groups, divided into two groups Lead (Ⅱ) acetate was added and stored frozen at -20 ° C. , And the mixture was divided into four groups, which were methanol, methanol, and methanol. The four groups were divided into 6 groups, 6-hydroxy-7,4'-dimethoxyflavone sample As a result, it was presumed that there was a component in the supernatant which was not dissolved in methanol.

Thin layer chromatography was used to concentrate the mixture. When methanol and chloroform were mixed at a ratio of 1: 19, the highest Rf value was found. The ratio of Rf value was large and divided into two components. This is presumed to be 6-hydroxy-7,4'-dimethoxyflavone.

It was confirmed that the supernatant was not dissolved in the methanol obtained from the thin layer chromatography. Then, the solution was filtered twice with methanol to remove crystals that might be present in the lower layer. The filtrate was taken twice at a ratio of the solvent obtained through thin layer chromatography. At the beginning, about 40 g of methanol and chloroform Was obtained in a ratio of 1: 19 in the ratio of 30 to 40 ml. In the second step, about 11 g of ethyl acetate and chloroform were collected in a beaker at a ratio of 1: 3 in a ratio of 1: 3. The difference between the amounts was not so large, and the same solvent was used, and 0.5 ml of each solution was received by a test tube, and the width of the point was increased more than when receiving with a 10 ml beaker. The sections came in two, concentrated and identified as 6-hydroxy-7,4'-dimethoxyflavone.

[Table 1]

[Table 2]

Example 2: 6 - hydroxy -7,4'- dimethoxyflavone  Identification of structure of compound

The final product of 6-hydroxy-7,4'-dimethoxyflavone, which was finally obtained in column chromatography, was dissolved in DMSO and analyzed by 6-hydroxy-7,4'-dimethoxyflavone by 1 H- .

Example  3: From black ginger  The isolated 6- hydroxy -7,4'- dimethoxyflavone  Investigation of biological activity of diabetic therapeutic substance

The normal group showed an obvious increase in body weight because of the use of growing animals. However, the group treated with STZ showed a significant difference in weight before STZ, and it was found that the increase in weight was less obvious than the other group when it became 7 days later. It can be seen that the degree of increase in body weight is small. It has been pointed out that STZ treatment is a typical phenomenon occurring in diabetes-induced rats in a short time, which is caused by abnormality of saccharide metabolism.

The changes of blood glucose according to administration of 6-hydroxy-7,4'-dimethoxyflavone are summarized in FIG. Changes in blood glucose levels in the normal group are not much different from those in the normal group. On the other hand, it can be seen that there is not a large difference between the STZ treatment on the 0th day (the 5th day after the STZ re-administration) and the treatment with the STZ and the drug. The first dose of the drug is 6-hydroxy-7,4'-dimethoxyflavone. In the third week, 6-hydroxy-7,4'-dimethoxyflavone lowers blood sugar in diabetes. And the STZ diabetic group also shows some natural fallen adaptation to some degree.

Compared to the STZ diabetic group, the clearance of the drug treatment was significantly lower than that of the STZ diabetic group. The 6-hydroxy-7,4'-dimethoxyflavone level was not significantly higher than the normal level, 6-hydroxy-7,4'-dimethoxyflavone seems to be effective in diabetes even at low doses.

Approximately 2.5 ml of supernatant is obtained after about 2 hours of blood obtained by dissecting mice and 2000 ml of blood for 10 minutes. As a result of analysis, C-peptide was not significantly different according to administration of 6-hydroxy-7,4'-dimethoxyflavone. The results of C-peptide measurement according to administration of 6-hydroxy-7,4'-dimethoxyflavone are summarized in FIG.

Insulin is low in diabetes. The results showed that the STZ group had the lowest value and the drug group was slightly higher than the normal group. As a result, it was confirmed that 6-hydroxy-7,4'-dimethoxyflavone has a strong therapeutic effect on diabetes. The results of insulin measurement according to administration of 6-hydroxy-7,4'-dimethoxyflavone are summarized in FIG.

Triglyceride is elevated when there is diabetes. The results showed that the STZ group had the highest value, and the drug group showed a slight decrease by concentration, and the normal group showed the lowest value. The results of the measurement of the triglyceride according to the administration of 6-hydroxy-7,4'-dimethoxyflavone are summarized in FIG.

Claims (9)

6-hydroxy-7,4'-dimethoxyflavone or a pharmaceutically acceptable salt thereof for the prevention and treatment of diabetes.
The pharmaceutical composition for preventing or treating diabetes according to claim 1, wherein the 6-hydroxy-7,4'-dimethoxyflavone is obtained from black ginger extract.
The pharmaceutical composition for preventing or treating diabetes according to claim 2, wherein the black ginger extract is extracted with an organic solvent.
[Claim 4] The pharmaceutical composition for preventing or treating diabetes according to claim 3, wherein the black ginger extract is obtained from the ethylacetate fraction by further performing an ethyl acetate fraction after the organic solvent extraction.
The pharmaceutical composition for preventing or treating diabetes according to claim 3, wherein the organic solvent is a C1 to C4 alcohol.
The pharmaceutical composition for preventing or treating diabetes according to claim 5, wherein the C1 to C4 alcohol is methanol.
The pharmaceutical composition for preventing or treating diabetes according to claim 1, wherein a single dose of the composition is such that 6-hydroxy-7,4'-dimethoxyflavone is contained in an amount of 50 to 200 mg per 1 kg of body weight of the subject to be administered.
6-hydroxy-7,4'-dimethoxyflavone.
Extracting black ginger with methanol;
Suspending the methanol extract in water; And
Adding ethyl acetate to the suspension and obtaining an ethyl acetate fraction;
To extract 6-hydroxy-7,4'-dimethoxyflavone from black ginger.

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