JP5156298B2 - Muscle cell sugar transport promoting composition - Google Patents

Description

  The present invention relates to a composition for promoting muscle cell sugar transport, and more particularly to a composition for promoting muscle cell sugar transport, characterized by containing cardamonin as an active ingredient.

Muscles (skeletal muscles, myocardium), especially skeletal muscles, are the largest organs in the human body that occupy about half of their body weight and are places where most of the glucose (sugar) obtained from food is stored or used as energy. Therefore, it is known that sugar uptake in skeletal muscle greatly regulates blood glucose concentration (blood glucose level).
When the absorbed sugar enters the portal vein after meals, additional secretion of insulin occurs instantaneously from the pancreas, and the liver switches its action to suppress the release of sugar and conversely promote the uptake of sugar.
On the other hand, excessive caloric intake due to overeating or lack of exercise results in a loss of energy balance, fat cell size or number increases with obesity, and excessive accumulation of neutral fat in hepatocytes and skeletal muscles. This results in an insulin resistance state in which the action of insulin that enhances muscle blood glucose uptake is reduced.
If the liver has insulin resistance, even if sugar flows into the liver after meals, sugar intake into the liver does not go well, and the amount of sugar released from the liver does not decrease, so the amount of sugar that flows into the blood In addition, if insulin resistance exists in the skeletal muscle on the side where sugar is used, the sugar is not taken into the skeletal muscle well, and the blood glucose level rapidly increased after eating falls into a state where it does not fall easily.
If this condition continues for a long time, the pancreas further increases insulin secretion and promotes glucose uptake in the blood. As a result, when the burden on the pancreas exceeds the limit, the insulin secretion function rapidly declines and diabetes occurs .

  As such a therapeutic / ameliorating composition for diabetes, a composition that promotes differentiation of preadipocytes into adipocytes and promotes glucose metabolism is known (Patent Document 1). In addition, it has been clarified that the amount of sugar uptake in skeletal muscle is decreased in diabetic patients (Non-Patent Documents 1 to 3), and screening for antidiabetic drugs using muscle cell sugar transport promoting action as an index is also possible. (Non-Patent Document 4).

On the other hand, cardamonin is contained in Alpinia katsumadai, Boesenbergia pandurata, etc. of Zingiberaceae, a kind of herbal medicine, and has antitumor activity, antimutagenicity, anti-inflammatory activity. Have been reported to have an antioxidant effect (Patent Document 2, Non-Patent Documents 5 to 7).
However, the action of cardamonin on skeletal muscle is not known.

Japanese Patent No. 3069686 JP-A-6-279270 The Journal of clinical investigation 76, 149-155 (1985) Diabetologia 43, 821-835 (2000) Biochemical Society transactions 33, 354-357 (2005) J. Med. Chem. 1998 Nov 5; 41 (23): 4556-4566 J. Agric. Food Chem., 49 (6), 3046-3050, 2001 British Journal of Pharmacology (2006) 149, 188-198 Z. Naturforsch. 59c, 811-815 (2004)

  An object of this invention is to provide the composition which accelerates | stimulates the glucose transport effect | action by a muscle cell (skeletal muscle or a cardiac muscle).

As a result of intensive studies in order to solve the above problems, the present inventors have found that skeletal muscle cells stimulated by cardamonin have an excellent blood glucose uptake effect. Furthermore, the present inventors have also found that the composition for promoting muscle cell sugar transport containing cardamonin of the present invention is useful for the prevention or treatment of diabetes.
This preventive or therapeutic effect of diabetes is based on the action of cardamonin on muscle cells and as a result, it works directly into the muscles to promote glucose transport. The blood glucose level at the time can also be lowered. Therefore, the present invention is also a hypoglycemic agent or an agent for preventing or treating diabetes, which contains cardamonin and has a muscle cell sugar transport promoting action.
In addition, examples of tissues that promote glucose uptake in an insulin-dependent manner include fat and skeletal muscle, but when adipocyte sugar uptake is promoted, adipocytes enlarge and secrete adipocytokines that cause insulin resistance. It is not an essential solution for the treatment of diabetes because it leads to failure and enhancement. However, since skeletal muscle is reported to take up 85% of glucose absorbed in the body, it is known to be the most important organ for lowering blood glucose level. unknown. Therefore, it can be said that the method of promoting the sugar transport of muscle cells is the most effective method for the prevention or treatment of blood glucose lowering and diabetes not related to food intake. Based on these findings, the inventors of the present invention have further advanced research and have completed the present invention.

That is, the present invention
[1] A composition for promoting muscle cell sugar transport, comprising cardamonin as an active ingredient,
[2] The composition according to [1], which is a medicine,
[3] The composition according to the above [2], which is a hypoglycemic agent,
[4] The composition according to the above [2], which is a preventive or therapeutic agent for diabetes,
[5] The composition according to [1], which is a food, and [6] the composition according to [1], which is a food material.

  The composition of the present invention can promote glucose transport in muscle cells (skeletal muscle or cardiac muscle). When a person or animal ingests the composition, medicine or food of the present invention, sugar transport to muscle cells (skeletal muscle or myocardium) is promoted, blood glucose level can be lowered, and diabetes is prevented or treated. Can do.

Cardamomonin used in the present invention is contained in Zingiberaceae, a kind of herbal medicine, such as Alpinia katsumadai, Boesenbergia pandurata, and the following formula [I]:
It is 2 ', 4'-dihydroxy-6'-methoxychalcone (2', 4'-dihydroxy-6'-methoxy chalcone) shown by these.

Cardamomonin used in the present invention can be extracted from a plant containing it, using polar organic solvents (methanol, ethanol, etc.), water, nonpolar organic solvents, etc., and used in the form of the resulting extract It is also possible to use a chemically synthesized product. The extract can be used as it is in the composition of the present invention, or may be further used after being subjected to processing such as drying, powdering, granulating, and solution-forming by a conventional method.
The amount of cardamonin added to the composition of the present invention is such that, for example, when the composition is used as a beverage or the like, cardamonin is contained in an amount of about 0.00001 to 10% by weight, preferably about 0.0001 to 1% by weight. It is desirable to set. In addition, when the composition is used for oral administration in the form of a solid, powder, granule, paste or the like as a food, food material, or pharmaceutical product, it contains about 0.01% by weight or more of cardamonin. The upper limit is 100% by weight, that is, cardamonin may be used alone. Further, when the composition is used for parenteral administration, it is desirable to set it so that cardamonin is contained at about 0.00001 to 10% by weight, preferably about 0.0001 to 1% by weight. In the present specification, the term “ingestion” includes administration.

  In the present invention, the cardamonin may be used as it is as the composition for promoting the muscle cell sugar transport of the present invention, and other physiologically active substances may be blended. For example, excipients, pH adjusters, cooling agents, suspending agents, thickeners, solubilizers, disintegrating agents, binders, lubricants, antioxidants, coating agents, coloring agents, flavoring agents , A sweetener, a surfactant, a plasticizer, a gum base, a gelling agent, a fragrance and the like.

  Examples of the excipient include sugar alcohols such as D-sorbitol, D-mannitol or xylitol, sugars such as glucose, sucrose, lactose or fructose, crystalline cellulose, carmellose sodium, calcium hydrogen phosphate, wheat starch, rice starch. Corn starch, potato starch, dextrin, β-cyclodextrin, light silicic anhydride, titanium oxide, magnesium aluminate metasilicate, and the like.

  Examples of the pH adjuster include acetate, propionate, ammonium salt and the like. Examples of the refreshing agent include l-menthol or mint water. Examples of the suspending agent include kaolin, carmellose sodium, xanthan gum, methylcellulose, and tragacanth. Examples of the thickener include xanthan gum, tragacanth, methylcellulose, and dextrin. Examples of the solubilizer include ethanol, sucrose fatty acid ester, macrogol and the like. Examples of the disintegrant include low-substituted hydroxypropylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, hydroxypropyl starch, and partially pregelatinized starch.

  Examples of the binder include methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, gelatin, gum arabic, ethylcellulose, polyvinyl alcohol, pullulan, pregelatinized starch, agar, tragacanth, sodium alginate, or propylene glycol alginate. Can be mentioned. Examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, polyoxyl stearate, cetanol, talc, hydrogenated oil, sucrose fatty acid ester, glycerin fatty acid ester, lecithin, dimethylpolysiloxane, beeswax and white beeswax. Can be mentioned. Examples of the antioxidant include dibutylhydroxytoluene (BHT), propyl gallate, butylhydroxyanisole (BHA), tocopherol, and citric acid. Examples of the coating agent include hydroxypropylmethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carboxymethylethylcellulose, cellulose acetate phthalate, polyvinyl acetal diethylaminoacetate, aminoalkyl methacrylate. Copolymer, hydroxypropyl methylcellulose acetate succinate, methacrylic acid copolymer, polyvinyl acetate diethylaminoacetate or shellac.

  Examples of the colorant include turmeric extract, riboflavin, titanium oxide, or carotene solution. Examples of the flavoring agent include organic acids, and specifically, citric acid, malic acid, tartaric acid, succinic acid, lactic acid, acetic acid, phosphoric acid, maleic acid, gluconic acid, aspartic acid, adipic acid, Examples thereof include glutamic acid and fumaric acid. Examples of the sweetener include fructose-glucose liquid sugar, sugar, starch syrup (reduced starch syrup), glucose, stevia, licorice and the like. Examples of the surfactant include polyoxyethylene hydrogenated castor oil, glyceryl monostearate, sorbitan monostearate, sorbitan monolaurate, polyoxyethylene polyoxypropylene, polysorbates, sodium lauryl sulfate, macrogol or sucrose fatty acid. Examples include esters. Examples of the plasticizer include triethyl citrate, polyethylene glycol, triacetin, and cetanol. Examples of the gum base include ester gum, chicle or beeswax. Examples of the gelling agent include agar, carrageenan, fur celerane, gellan gum, pectin, starch, xanthan gum, locust bean gum, guar gum, tara gum, tamarind seed gum, cassia gum or konjac mannan. Examples of the fragrances include natural fragrances such as animal fragrances and vegetable fragrances, and synthetic fragrances such as isolated fragrances and pure synthetic fragrances.

  The dosage form is appropriately selected according to the administration form or usage form, and is a known dosage form, for example, a solid dosage form such as powder, granule, tablet, capsule, troche, suppository or ointment, for example, injection. It can be processed into liquid dosage forms such as agents (solutions or suspensions), gels, sols and the like. The administration form is generally oral administration, but is not limited thereto.

  In the present invention, the composition containing cardamomonin may be a food, may be a food additive alone, or may be a pharmaceutical composition (a hypoglycemic agent or a preventive or therapeutic agent for diabetes). For example, it is used as a food production raw material by a known means such as adding a composition containing cardamonin to a food, its foodstuff, or its food intermediate material.

  Examples of the food include confectionery (for example, snacks including potato chips, baked confectionery such as biscuits or cookies, chocolate, gum, gummy, candy, etc.), desserts (for example, pudding, jelly, yogurt or ice cream). In addition to taste foods such as noodles (for example, buckwheat, udon, ramen or pasta), food equivalent to staple foods such as cereals (for example, corn flakes or oatmeal), seasoned food (for example, soup, curry or stew) Processed agricultural products (eg jams), milk oil foods (eg spreads or cheese), health foods (eg protein or fiber), calorie-adjusted foods, non-alcoholic beverages (eg soybean roasted tea drinks, grain teas, coffee drinks) , Tea drinks, green tea drinks, Tea drinks, matcha tea drinks, vegetable juice drinks, fruit juice drinks (orange juice, grapefruit juice etc.), soft drinks etc., or alcoholic drinks (eg beer, wine, sake, sparkling wine, plum wine, whiskey, brandy, shochu, vodka Lamb, gin, liqueur or cocktail).

  Furthermore, carbohydrates, proteins, herbal medicines, minerals, vitamins, amino acids, and the like may be appropriately blended in the muscle cell sugar transport promoting composition, food or food additive.

  Examples of the carbohydrate include polysaccharides such as starch or corn starch, and other saccharides such as dextrin, sucrose, glucose or fructose. Examples of the protein include a combination of animal protein and plant protein, and more specifically, milk protein (milk, skim milk powder, etc.), brown rice germ oil, wheat-derived gluten, corn-derived zein, Examples include soy protein or egg protein. Examples of the herbal medicine include valerian, toki, glaze, peony, ginseng and the like. Examples of the minerals include calcium, magnesium, iron, sodium, potassium, zinc, copper, chromium, selenium, manganese, molybdenum, nickel, and vanadium. Examples of the vitamins include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, niacin, nicotinic acid, lipoic acid, biotin, ubiquinone, pantothenic acid or folic acid. Etc. Examples of the amino acid include glutamine, leucine, isoleucine, valine, tryptophan, phenylalanine, lysine, threonine, methionine, histidine, glycine, alanine, serine, aspartic acid, glutamic acid, asparagine, arginine, cystine, cysteine, tyrosine, proline or hydroxy And proline.

  When the pharmaceutical composition (hypoglycemic agent or diabetes preventive or therapeutic agent) is prepared, for example, as an injection, in addition to a solvent (eg, purified water), a dispersant (eg, Tween 80 (Atlas powder) Manufactured by the United States), HCO60 (manufactured by Nikko Chemicals), polyethylene glycol, carboxymethylcellulose, sodium alginate, etc., preservatives (methylparaben, propylparaben, benzyl alcohol, etc.), isotonic agents (sodium chloride, mannitol, sorbitol, glucose) Etc.) can be used as appropriate.

  The intake of the composition of the present invention varies depending on the sex, age, health condition, etc. of the person who takes it. However, in the case of oral intake, considering the absorption rate, about 0 per day for an adult as cardamonin. It is desirable to set the dose to be in the range of about 0.01 mg to 5000 mg, preferably about 0.1 mg to 1000 mg, more preferably about 1 mg to 200 mg, and the desired effect can be obtained. In the case of parenteral intake such as injections or infusions, it is set so that cardamonin is administered at about 0.001 mg to 2500 mg, preferably about 0.01 mg to 500 mg, more preferably about 0.1 mg to 100 mg. Is desirable. Moreover, the frequency | count of ingestion may be once per day, or may be multiple times. The cardamonin used in the present invention is itself obtained from an edible material, and it can be said that its safety is very high.

Hereinafter, the present invention will be described with reference to Examples and Production Examples, but the present invention is not limited thereto. The cardamonin used in the following Examples and Production Examples was cardamonin extracted and purified from Alpinia katsumadai by the following method.
Extraction was performed with 400 ml of methanol from 25 g of dried seeds of sawtooth, and 4.5 g of extract was obtained. The extract was partitioned with ethyl acetate / water, and the ethyl acetate layer was concentrated to dryness (yield 2.5 g). Next, fractionation was performed by silica gel column chromatography using ethyl acetate as the mobile phase (yield 1.5 g), and fractionation was further performed by silica gel column chromatography using chloroform as the mobile phase to obtain 617 mg of a fraction containing cardamomonin. . This was fractionated by reverse phase column chromatography (ODS, 50 μm, 120 A, 30 × 200 mm, methanol / water (70:30, v / v), 8 ml / min) to obtain 170.5 mg of cardamonin.

[Example 1]
(Preparation of L6 skeletal muscle cells)
L6 myoblasts were adjusted with α-MEM medium (containing 10% v / v fetal bovine serum) to a concentration of 2.5 × 10 ⁴ / ml, and placed on a 96-well tissue culture plate coated with collagen type I. After inoculating 200 μl per well and culturing for 2 days in a 5% carbon dioxide incubator at 37 ° C. and becoming confluent, the medium was changed to α-MEM medium (containing 2% v / v fetal calf serum). L6 skeletal muscle cells were prepared by culturing in a 5% carbon dioxide incubator at 5 ° C. for 5 days.

(Confirmation of glucose transport promoting action in L6 skeletal muscle cells)
Cardamonin was dissolved in dimethyl sulfoxide and diluted 100-fold in α-MEM medium (containing 2% v / v fetal calf serum) to prepare α-MEM medium containing 0, 10, and 37 μM cardamonin. The α-MEM medium containing cardamonin was added to each L6 skeletal muscle cell culture plate in an amount of 170 μl per well, and the plate was incubated for 4 hours in a 37 ° C. 5% carbon dioxide incubator. Next, Krebs-Ringer-phosphate-Hepes (KRRH) buffer (pH 7.4, 20 mM Hepes, 5 mM KH ₂ PO ₄ , 1 mM MgSO ₄ , 1 mM CaCl ₂ , 136 mM NaCl, 4.7 mM KCl containing 0.1% BSA ) Was washed twice with 200 μl per well, and 100 μl per well of KRPH buffer (pH 7.4) containing 1 mM 2-deoxyglucose (2-DG) and 0.1% BSA was added. Incubated for 20 minutes in a 5% carbon dioxide incubator. Further, after washing twice with 200 μl per well with KRPH buffer (pH 7.4) containing 0.1% BSA, 25 μl of 0.1N NaOH was added per well, and the plate was frozen in a freezer. . After freezing, the plate is taken out of the freezer, returned to room temperature, heated in a hot water bath at 85 ° C. for 40 minutes, allowed to cool to room temperature, 25 μl of 0.1N HCl is added per well, and then 150 mM. A sample to which 25 μl of triethanolamine buffer (pH 8.1) was added per well was used as a sample. By quantifying 2-DG in each sample as follows and comparing it with the 2-DG uptake of control (α-MEM medium (containing 2% v / v fetal bovine serum only)), the effect of promoting sugar transport I investigated.

(Quantification of 2-deoxyglucose)
The sample was added to a 96-well tissue culture plate at 50 μl per well, and assay cocktail 150 μl (50 mM triethanolamine buffer (pH 8.1), 50 mM KCl, 0.5 mM MgCl ₂ , 0.02% BSA, 670 μM ATP, 120 μM). NADP +, 25 μM resazurin sodium, 5.5 units / ml hexokinase, 16 units / ml glucose-6-phosphate dehydrogenase, 1 units / ml diaphorase), stirred with a microplate mixer, and cultured at 37 ° C. in 5% carbon dioxide gas After incubation for 90 minutes in a vessel, the quantification was performed by measuring the fluorescence value (excitation wavelength: 530 nm, measurement wavelength: 590 nm) with a microplate reader.

In addition, as a control compound, alpinetine, naringenin and naringenin chalcone having a structure similar to cardamonin were used, and the sugar transport promoting action was examined in the same manner as described above using 2-DG uptake as an index. The structure of the reference compound is as follows:
Alpinetin (Alpinetin, 5-methoxy-7-hydroxy favanone)
Naringenin (5,7,4'-trihydroxy favanone)
Naringenin chalcone (5,7,4'-trihydroxy chalcone)

(result)
The sugar transport promoting action (2-DG uptake amount) of cardamonin and the control compound is as shown in Table 1 and FIG. In FIG. 1, * indicates that there was a significant difference at a significance level of 5%, and ** indicates that there was a significant difference at a significance level of 1%.
*) The 2-DG uptake amount was displayed as a relative value (%) when the 2-DG uptake amount was 100 when no sample compound was added.

  From the above results, when skeletal muscle cells were stimulated with cardamonin, flavanones such as alpinetine and naringenin and derivatives of naringenin, and when skeletal muscle cells were stimulated with naringenin chalcone, a structural analog of cardamonin As compared with the above, it was confirmed that the uptake amount of 2-deoxyglucose (2-DG) increased significantly, and cardamonin promoted sugar transport in muscle cells.

[Production Example 1]
Each raw material was mixed well in the blending amounts shown in Table 2 below, and then tableted to produce 1 g of 1 tablet (containing cardamomonin 50 mg / tablet).

[Production Example 2]
Soft drinks were produced with the blending amounts shown in Table 3 below.

[Production Example 3]
Soft capsules were produced with the blending amounts shown in Table 4 below.

[Production Example 4]
Yogurt was produced with the blending amounts shown in Table 5 below.

[Production Example 5]
Candy was manufactured with the compounding amounts shown in Table 6 below.

[Production Example 6]
Gummy was produced with the blending amounts shown in Table 7 below.

[Production Example 7]
Gum was produced with the blending amounts shown in Table 8 below.

  According to the present invention, a composition having an excellent sugar transport promoting action in muscle cells can be provided.

FIG. 1 shows the amount of 2-DG taken up by skeletal muscle cells when stimulated with cardamomon, alpinetine, naringenin or naringenin chalcone of Example 1.

Claims (4)

  A composition for promoting muscle cell sugar transport, comprising cardamonin as an active ingredient.   The composition according to claim 1, which is a medicine.   The composition according to claim 2, which is a hypoglycemic agent.   The composition according to claim 2, which is a preventive or therapeutic agent for diabetes.