JP2013245213A - Clathrate of eriobotrya japonica leaf culture extract and cyclodextrin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a clathrate of an Eriobotrya japonica leaf culture extract and a cyclodextrin.SOLUTION: A clathrate of an Eriobotrya japonica leaf culture extract and a cyclodextrin is provided. A blood glucose level rise inhibitor, functional food, and cosmetics each including the clathrate are also provided.

Description

  The present invention relates to inclusions of loquat tissue, in particular loquat leaf culture extract and cyclodextrin.

Cyclodextrin (CD) is a cyclic oligosaccharide obtained by allowing cyclodextrin glucanotransferase to act on corn or potato starch, in which glucose is α-1,4-linked. CD has hydrophilicity on the outer side of the ring and hydrophobicity on the inner side of the ring, and has a property (inclusion action) of encapsulating various molecules mainly including a fat-soluble substance in the hydrophobic cavity. By this inclusion action, it is possible to improve the characteristics of various pharmacologically active substances, and improvements in stability, sustained release, solubilization, powdering, hygroscopicity and the like are expected (Non-Patent Document 1).
In addition, patent applications relating to various inclusion compounds have been made for the purpose of improving the water solubility, stability, physiological activity and the like of the target substances (Patent Documents 1 to 7).

On the other hand, corosolic acid is a triterpene present in plants and can be obtained mainly from banaba (Lagerstroemia speciosa). Corosolic acid is known to lower blood glucose levels and suppress re-elevation of blood glucose levels (Non-patent Document 2: Antidiabetic activity of a standardized extract (Glucosol) from Lagerstroemia speciosa leaves in Type II diabetics. A dose- Judy WV. et al., Journal of Ethnopharmacology 87: 115 117, 2003), and attempts have been made to extract corosolic acid in large amounts from loquat cell suspension culture (Patent Document 8). Furthermore, a blood sugar level increase inhibitor containing corosolic acid and / or other triterpenes and cyclodextrins is also known (Patent Document 9). In Patent Document 9, corosolic acid and / or other triterpenes are extracted from loquat. It can be isolated from the extract.
Furthermore, an attempt to produce a skin cosmetic containing corosolic acid has also been made (Patent Document 10).

  However, since the above-mentioned blood sugar level increase inhibitor and cosmetics need to use isolated and purified corosolic acid and / or other triterpenes, which are active ingredients, it requires a processing device and takes time for processing work. Cost. In addition, isolated triterpenes, particularly tormic acid, have low oxidation stability, and there is a problem that deterioration occurs in handling in the atmosphere.

Japanese Patent No.4859006 JP 2010-24240 JP 2012-19695 Gazette JP-A-8-259829 JP 2010-174105 A JP-A-62-267261 JP 2010-126492 A Special Publication 2009-500013 Publication JP 2006-347967 JP 2006-265232 A

Daisuke Nakata, Keiji Terao, Vitamins, Vol. 84, No. 2 (February) 2010, 61-70 Judy WV. Et al., Journal of Ethnopharmacology 87: 115, 117, 2003

  It is not clear whether a loquat tissue culture extract that is easily prepared without isolating and purifying corosolic acid has a predetermined pharmacological effect. In addition, the culture extract of loquat tissue was encapsulated with cyclodextrin, whether the inclusion has oxidative stability, and further, in the human intestinal tract, solubilization in the digestive tract is promoted and absorbability It is not clear whether or not will increase. Therefore, an object of the present invention is to provide an inclusion product of loquat tissue culture extract and cyclodextrin. Another object of the present invention is to provide a blood sugar level increase inhibitor comprising the inclusion product or loquat tissue culture extract.

  As a result of intensive studies to solve the above-mentioned problems, the present inventor has found that loquat tissue culture extract suppresses an increase in blood glucose level, and inclusions of loquat tissue culture extract and cyclodextrin are high in the intestinal tract. As a result, it has been found that it has an action of lowering blood glucose level, and the present invention has been completed.

That is, the present invention is as follows.
(1) An inclusion product of a culture extract of loquat tissue and cyclodextrin or a derivative thereof.
(2) The inclusion according to (1), wherein the loquat tissue is loquat leaves.
(3) Corosolic acid, maslinic acid, tormentic acid, ursolic acid, 2α, 19α-dihydroxy-3-oxours-12-en-28-oic acid, oleanolic acid and fupendic acid and their pharmaceutically acceptable An inclusion product of at least one selected from the group consisting of salts and cyclodextrin or a derivative thereof.
(4) The clathrate according to any one of (1) to (3), wherein the water solubility is improved more than the water solubility of the non-inclusion loquat leaf culture extract.
(5) The inclusion according to (4), wherein the intestinal absorptivity is improved more than the intestinal absorptivity of the non-encapsulated loquat leaf culture extract by improving water solubility.
(6) As described in (5), the intestinal absorbability is improved, so that the amount of components transferred in the inclusion is increased in blood than the amount of components in the non-inclusion loquat leaf culture extract. Inclusions.
(7) The clathrate according to (6), wherein a component in the clathrate is transferred into the blood, thereby exhibiting an effect of suppressing an increase in blood sugar level.
(8) A blood sugar level increase inhibitor comprising a culture extract of loquat tissue or an inclusion product according to any one of (1) to (7).
(9) The inhibitor according to (8), wherein the loquat tissue is loquat leaves.
(10) The inhibitor according to (8) or (9), wherein the blood glucose level is a fasting blood glucose level.
(11) The inhibitor according to (10), which is used for a human having a fasting blood glucose level of at least 100 mg / dl.

  According to the present invention, an inclusion product of loquat tissue culture extract and cyclodextrin is provided. The present invention also provides a blood sugar level increase inhibitor containing loquat tissue culture extract.

  In the inclusion product of the present invention, the water solubility in the digestive tract is improved more than the water solubility of the non-inclusion loquat leaf culture extract, thereby improving the absorbability in the intestinal tract. And the amount of the active ingredient in the inclusion contained in the blood is higher than the amount of the active ingredient in the non-inclusion loquat leaf culture extract, and as a result, the blood sugar level, particularly the fasting blood sugar level is lowered. Has an effect. Furthermore, the clathrate of the present invention has oxidative stability. Therefore, the clathrate of the present invention is extremely useful for functional foods for the purpose of suppressing an increase in blood glucose level.

It is a figure which shows that loquat leaf culture extract reduces a fasting blood glucose level.

Hereinafter, the present invention will be described in detail.
1. Outline The present invention is a culture extract of loquat tissue itself, or an inclusion product of a culture extract of loquat tissue and cyclodextrin, and can be used as a blood sugar level increase inhibitor, a functional food and the like.

  In this example, a loquat leaf culture extract was prepared, encapsulated and used for humans, and it was found that the loquat leaf culture extract has an effect of suppressing an increase in blood glucose level. However, since loquat leaf culture extract is insoluble or hardly soluble in water, it is necessary to eat a large amount of extract in order to obtain intestinal absorbability. This not only increases the manufacturing cost but also burdens consumers.

  Therefore, in the present invention, the loquat leaf culture extract was encapsulated with cyclodextrin in order to increase the water solubility of the culture extract and improve the intestinal absorbability. As a result, the present inventors have found that even a small amount of loquat leaf culture extract increases the amount of active ingredients transferred into the blood and suppresses an increase in blood glucose level. Thereby, the manufacturing cost of a loquat leaf culture extract can be held down and a consumer's burden is also reduced.

In the present invention, a loquat tissue culture extract-cyclodextrin inclusion product is prepared by mixing cyclodextrin and loquat tissue culture extract. Loquat tissue culture extracts (eg loquat leaf culture extract) do not dissolve in water, but cyclodextrin inclusions dissolve in water and water-alcohol. Alternatively, the dispersibility is good in water or an aqueous medium. In the present invention, an inclusion can be prepared by using any of α, β, and γ-cyclodextrin, but it is preferable in view of the highest water solubility of the γ-cyclodextrin inclusion.
In addition, the clathrate of the present invention has improved intestinal absorbability, and stability against heat, oxygen, pH, etc. is improved, and improvement in efficacy and efficacy resulting from this is expected.

2. Loquat tissue culture extract The plant used in the present invention is loquat (Eriobotrya japonica).
Biwa is an evergreen Takagi of the Rosaceae family, the fruit part is edible, and the leaf part is used for the treatment of gastrointestinal weakness, neuralgia, diarrhea and the like. Loquat is native to Japan, Taiwan, China, etc., and can be easily obtained from these regions. The tissue of loquat that can be used as the raw material for extraction is not particularly limited, for example, leaf parts, branch parts, bark parts, trunk parts, stem parts, fruit parts, seed parts, flower parts, etc. Although the mixture of these site | parts etc. are mentioned, It is preferable to use a leaf part among these.

The loquat tissue culture extract used in the present invention, particularly the loquat leaf culture extract, is an extract from the culture and can be obtained by the following method.
First, callus is induced from loquat plant tissue, and a cell line for suspension culture is prepared from this callus. Next, this cell line is suspended and cultured in a liquid medium. Then, a culture extract is extracted from the obtained suspension culture. The culture extract can be obtained by an extraction method generally used for plant extraction.
Callus induction of loquat tissue can be performed with reference to a known technique (for example, JP-T-2009-500013).

For example, when loquat leaves are used as the tissue, the loquat leaves are sterilized and then cultured on a nutrient medium containing a plant growth regulator. As the nutrient medium, a medium widely used for culturing plant cells, for example, Murashige-Skoog medium (MS medium), Gamborg medium (B5 medium) and the like can be used. If necessary, various additives can be added to the medium, or some of the components can be removed for use. Examples of the additive include natural auxin such as indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), α-naphthalene acetic acid (NAA), and the like.
The preferred growth temperature for callus induction is 20-30 ° C, preferably 22-27 ° C.

  After the callus has grown sufficiently, the callus is transferred to a liquid medium and subjected to liquid suspension culture to induce a suspension culture cell line. As the medium used for liquid culture, a medium having the same composition as that of the medium used at the time of callus induction, excluding solidified components such as agar, can be used. Suspension culture can be performed by a known method such as batch culture, continuous culture, fed-batch culture, or the like, and the culture conditions can also be the same as the callus induction.

After suspension culture, the cells are collected by centrifugation or the like, and the loquat tissue culture extract is extracted using the collected cells.
As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water, a hydrophilic organic solvent, and the like. These may be used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. Is preferred.

  Examples of water that can be used as the extraction solvent include pure water, tap water, and the like, and those subjected to various treatments are included. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. The water that can be used as the extraction solvent in the present invention includes physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of the hydrophilic organic solvent that can be used as the extraction solvent include lower aliphatic alcohols having 1 to 5 carbon atoms (eg, methanol, ethanol, isopropanol, butanol), lower aliphatic ketones (acetone, methyl ethyl ketone, etc.), and carbon. Examples thereof include polyhydric alcohols of 2 to 5 (1,3-butylene glycol, propylene glycol, glycerin and the like), and ethanol is preferable in consideration of use for food.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 0 to 50 parts by mass of water with respect to 50 to 100 parts by mass of the lower aliphatic alcohol, and water and a lower aliphatic ketone. Is preferably mixed with 50 to 100 parts by mass of a lower aliphatic ketone in an amount of 0 to 50 parts by weight of water, and when a mixture of water and a polyhydric alcohol is used. It is preferable to mix 0 to 30 parts by mass of water with 70 to 100 parts by mass of the polyhydric alcohol.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent in an amount of 0.1 to 10 times the extraction raw material (mass ratio), the soluble components are extracted at room temperature or under reflux heating, and then the extraction residue is removed by filtration. Can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

In the loquat tissue (eg loquat leaf) culture extract obtained in the present invention, triterpene is contained at a ratio of about 80 to 90%. Examples of triterpenes include tormentic acid, corosolic acid, maslinic acid, ursolic acid, 2α, 19α-dihydroxy-3-oxours-12-ene-28-oic acid, oleanolic acid, hupenic acid and the like. In addition, the culture extract contains plant sterols, fatty acids, phospholipids, and the like.
This culture extract is included by using cyclodextrin described later.

  The culture extract of loquat tissue contains various triterpenes, and these are used in the form of a composition without isolation.In the present invention, tormentic acid, corosolic acid, maslinic acid, ursolic acid, At least one triterpene selected from the group consisting of 2α, 19α-dihydroxy-3-oxours-12-ene-28-oic acid, oleanolic acid and fupendic acid, or a pharmaceutically acceptable salt thereof alone or as appropriate It can also be included in combination.

  In the case of a triterpene which is unstable with a single substance, it can be stabilized by making a mixture with other triterpenes. In this regard, in the present invention, it was confirmed that the oxidation stability of tormentic acid in the culture extract was improved. Therefore, triterpene (tormentic acid) is present in the culture extract, so that the oxidative stability is improved as compared with the case where it is isolated.

The triterpene single item can be isolated from loquat tissue culture extract using a method such as HPLC.
Of the above-mentioned triterpenes, corosolic acid, ursolic acid, and oleanolic acid are commercially available and can be purchased (Corporation Research Institute, Wako Pure Chemical Industries, Ltd., etc.).

  As a pharmaceutically acceptable salt, a salt with an alkali can be used. For example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, trimethylamine, triethylamine, pyridine, picoline, dicyclohexylamine, N, N'-dibenzylethylenediamine, arginine, lysine, etc. Organic base salts, ammonium salts, and the like.

3. Inclusion Product The production of the loquat tissue culture extract-cyclodextrin inclusion product can be performed using a known method.
Examples of the cyclodextrin compound include α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin, and among them, γ-cyclodextrin is preferable.

  In the present invention, a derivative of cyclodextrin can also be used. Cyclodextrin derivatives include chemically modified products such as hydroxypropylated, acetylated, triacetylated, methylated, monochlorotriazinated cyclodextrins. These derivatives can also be used by purchasing, for example, commercially available products (Cyclochem).

  As a method for inclusion, for example, a clathrate can be produced by forming a clathrate by a kneading method, a liquid phase mixing method, a solvent evaporation method or a freeze-drying method, and then drying, preferably a kneading method. (For example, Japanese Patent No. 4485006).

  The kneading method is performed according to a conventional method, for example, using a kneader, a mortar or the like, adding loquat leaf culture extract, cyclodextrin, and an appropriate amount of water and / or an organic solvent, and kneading until a paste is formed. By doing.

  As the organic solvent to be used, ethanol, methanol, THF (tetrahydrofuran), acetone, ethyl acetate, hexane, acetonitrile, chloroform, dichloromethane, and the like can be used, and ethanol is preferable in consideration of use for food. .

  The liquid phase mixing method is performed by adding a loquat leaf culture extract, cyclodextrin, an organic solvent and / or water in a container such as a flask or a reaction kettle and stirring vigorously in the aqueous phase according to a conventional method.

  The solvent evaporation method is performed by mixing loquat tissue culture extract, cyclodextrin, an organic solvent and / or water in an apparatus capable of reducing pressure, such as a rotary evaporator, and then distilling off the solvent under reduced pressure.

  The lyophilization method is performed by mixing the loquat tissue culture extract, cyclodextrin, organic solvent and / or water using a freeze dryer, for example, and then distilling off the solvent under reduced pressure.

  The mixing ratio of loquat tissue culture extract and cyclodextrin is, for example, 0.1 to 10 parts by mass of cyclodextrin with respect to 1 part by mass of loquat tissue culture extract, and the amount of water is not particularly limited. 0.001-1 mass part is preferable with respect to a mass part.

  The temperature condition for inclusion is preferably performed under a temperature condition in which the loquat tissue culture extract is not denatured, and is, for example, 4 to 50 ° C, preferably 4 to 25 ° C.

  In the present invention, water solubility can be improved by refining a loquat tissue culture extract and preparing a nanoparticle suspension which is an ultrafine particle preparation having an average particle size of 1 μm or less, preferably 100 nm or less. This method utilizes the fact that the dissolution rate of a solid is proportional to its surface area, and improves the dissolution rate by increasing its surface area by refining the solid drug. As the dissolution rate is improved, the intestinal absorbability is also improved.

Examples of the miniaturization method include a method using an airflow pulverizer (jet mill) used in a dry pulverization method, a supercritical freeze granulation method, a wet pulverization method, and the like. In addition, it can be miniaturized into nano-order particles using a nanomizer.
In addition, micelles can be formed to increase the water solubility of loquat tissue culture extract.

  When micelles have a hydrophilic part and a hydrophobic part in the molecule (amphiphile) dissolved in water, the hydrophilic group is removed outside at a certain concentration (critical micelle concentration). It is an aggregate made up of molecules.

  In the present invention, the culture extract can be micelled by treatment with a dispersing agent. Although a dispersing agent is not specifically limited, Surfactant, polymer | macromolecule, saccharides, alcohols, glyceride, an acid, a base, salts, etc. are mentioned. More preferably, the emulsifier used for food is good, for example, lecithin, lysolecithin, glycerin fatty acid ester, glycerin organic acid fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, polyoxyethylene Sorbitan fatty acid esters, calcium stearoyl lactate and the like can be used.

  The water solubility of the clathrate of the present invention can be confirmed by quantitatively determining the triterpene concentration in the loquat tissue culture extract in an aqueous solution by HPLC in addition to a general solubility measurement method. And when it dissolves in water at the time of solubility measurement, and the triterpene peak is confirmed in the HPLC chromatogram, the water-solubility of the inclusion product of the present invention is the non-inclusion loquat leaf culture extract (non-inclusion loquat leaf) It is judged that the water solubility of the cultured extract was improved.

  In addition, the intestinal absorbability of the inclusion of the present invention is determined, for example, by using Caco-2 cells (a cell line derived from human colon cancer), which is a model of intestinal epithelial cells, on the luminal side of the Caco-2 cell membrane. Confirm by quantifying the triterpene concentration in the solution permeated to the basement membrane side after adding a solution (for example, HBSS buffer, culture solution, artificial digestion solution) and leaving it for 2-3 hours. Can do. Since the permeability coefficient in Caco-2 cells is correlated with intestinal absorption and bioavailability, it can be used for screening of gastrointestinal membrane permeability of drugs. When the triterpene concentration determined by LC / MS is higher than when the non-inclusion loquat leaf culture extract is used, the inclusion product of the present invention has improved intestinal absorbability due to improved water solubility. Judging from the intestinal absorbability of the non-inclusion loquat leaf culture extract.

  The blood transferability of components (for example, each triterpene) in the inclusion product of the present invention can be confirmed, for example, by measuring the blood triterpene concentration after ingestion using LC / MS. Since blood contains many substances that interfere with LC / MS measurement, it is separated into serum by centrifugation, extracted from the serum with a solvent, further removed, and concentrated by triterpene. can do. And when the blood triterpene concentration is higher than when using the non-inclusion loquat leaf culture extract, the components in the inclusion of the present invention are improved in intestinal absorption, It is judged that the amount of blood transferred to the blood component has increased more than the amount of blood transferred to the component of the non-inclusion loquat leaf culture extract.

  The fact that the water-solubility of the clathrate of the present invention is improved, the intestinal absorbability of the clathrate of the present invention is improved, or the amount of components in the clathrate of the present invention that is transferred to blood is increased. This means that even if the tissue culture extract itself is not eaten in large quantities, the effect of suppressing the increase in blood glucose level can be exhibited in a small amount, and it can be provided to consumers at a small amount and at a low price.

4). Fasting blood glucose level inhibitor, postprandial blood glucose level increase inhibitor, food and cosmetics (1) Blood glucose level increase inhibitor The blood glucose level increase inhibitor of the present invention contains loquat tissue culture extract itself or the inclusion of the present invention. It is a waste. The blood glucose level includes postprandial blood glucose level and fasting blood glucose level, and the blood glucose level elevation inhibitor of the present invention can be applied to both blood glucose levels, but is preferably used for the purpose of suppressing the increase in fasting blood glucose level. The

The clathrate of the present invention is excellent in intestinal absorbability, and the intestinal absorbability of the sparingly soluble loquat tissue culture extract can be increased as compared with the non-clasping loquat leaf culture extract. As a result, an increase in blood glucose level can be suppressed by components such as corosolic acid contained in the loquat tissue culture extract.
As the blood glucose level elevation inhibitor of the present invention, the loquat tissue culture extract or inclusion product of the present invention can be used as it is, but usually a mixture prepared by mixing an appropriate additive can also be used.

  Examples of the additive include excipients, binders, lubricants, disintegrating agents, coloring agents, flavoring agents, emulsifiers, surfactants, solubilizers, suspending agents, etc. Examples include tonicity agents, buffering agents, preservatives, antioxidants, stabilizers, absorption promoters, and the like, and these can be used in appropriate combinations as desired.

  Examples of the dosage form of the blood sugar elevation inhibitor of the present invention include oral dosage forms such as powders, granules, pills, soft capsules, hard capsules, tablets, chewable agents, quick disintegrating agents, syrups, and liquids. These preparations can be prepared according to a conventional method.

  Additives that can be used for formulation include animal and vegetable oils such as soybean oil, safflower oil, olive oil, germ oil, sunflower oil, grape seed oil, beef tallow, sardine oil, polyethylene glycol, propylene glycol, Surfactant such as polyhydric alcohol such as glycerin and sorbitol, sorbitan fatty acid ester, sucrose fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, purified water, lactose, starch, crystalline cellulose, D-mannitol, lecithin, gum arabic Excipients such as sorbitol liquid and sugar liquid, other sweeteners, colorants, pH adjusters, and fragrances can be exemplified. Tablets and granules may be coated by a known method, or may be wrapped with a sol or gel substance.

When using a blood sugar elevation inhibitor, the administration method is preferably oral administration, and the subject of administration is not particularly limited, and can be used for the prevention of blood sugar elevation if it is a healthy person, If it is a test subject who has a high blood glucose level, it can aim at reducing a blood glucose level. For example, in the case of a subject having a high blood glucose level, it is used for a human having a fasting blood glucose level of at least 100 mg / dl. Preferably, it is used for a human having a fasting blood glucose level of 100 to 140 mg / dl, more preferably 100 to 126 mg / dl.
Moreover, the blood sugar level increase inhibitor of the present invention can suppress an increase in blood sugar level without lowering the blood sugar level below the normal value range, and thus without causing hypoglycemia.

The dosage of the blood glucose level elevation inhibitor of the present invention varies depending on the degree of symptoms, the age or sex of the patient or subject, the body weight, the timing of administration, the administration interval, etc., and is not particularly limited.
The dose when administering the loquat tissue (eg loquat leaf) culture extract of the present invention is usually at least 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg per day for an adult (body weight 60 kg), 160 mg, 170 mg, 180 mg, 190 mg or 200 mg, preferably 90 to 450 mg, more preferably 200 to 450 mg, particularly preferably 444 mg, which is usually administered once a day or twice to 3 times a day Can be administered separately.
Therefore, when the loquat tissue culture extract itself of the present invention is administered, the blood sugar level increase inhibitor of the present invention is orally administered to humans at least 90 mg of loquat leaf culture extract per day 1 to 3 times a day. It is used so that it may be used.

When administering the clathrate of the present invention, the amount of loquat tissue culture extract is usually at least 10 mg, 11 mg, 12 mg, 13 mg, 14 mg or 15 mg per day for an adult (body weight 60 kg), preferably 15 mg to 60 mg. More preferably, it is 20 to 40 mg, which can be usually administered once a day or divided into 2 to 3 times a day.
Therefore, the blood glucose level increase inhibitor of the present invention when administering the inclusion product of the present invention is such that at least 10 mg of loquat tissue culture extract per day is orally administered to humans 1 to 3 times a day. It is used.

Furthermore, the blood glucose level increase inhibitor of the present invention when each triterpene is clathrated alone or appropriately in combination and the clathrate is administered is at least 8.8 mg of triterpene per day for an adult (body weight 60 kg). 9.6 mg, 10.5 mg, 11.4 mg, 12.3 mg or 13.1 mg, preferably 13.1 mg to 52.5 mg, more preferably 17.5 to 35.0 mg, which is usually administered once a day, or 2 times a day It can be divided into 3 to 3 doses.
Therefore, in the case of administering the clathrate of the present invention, at least 8.8 mg of triterpene (single product or combination) per day is orally administered to humans 1 to 3 times a day. It is used so that it may be used.

(2) Food The food of the present invention contains the loquat tissue culture extract itself or the inclusion product of the present invention, and is used as a functional food, supplement, etc. especially for the purpose of suppressing an increase in fasting blood glucose level. The
Examples of the form of food containing the inclusion product of the present invention (particularly functional food) include supplements (powder, granules, soft capsules, hard capsules, tablets, chewable tablets, quick-disintegrating tablets). , Beverages (tea, carbonated beverages, lactic acid beverages, sports beverages, etc.), confectionery (gummy, jelly, gum, chocolate, cookies, candy, etc.), oil, fat and oil foods (mayonnaise, dressing, butter, cream, margarine, etc.), seasonings Food (ketchup, sauce, etc.), liquid food, dairy products (milk, yogurt, cheese, etc.), breads, noodles (noodles, soba, ramen, pasta, fried noodles, kishimen, samen, cold wheat, rice noodles, etc.) . However, it is not limited to these forms.

As the functional food containing the inclusion product of the present invention, various nutrients, various vitamins (vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin E, etc.) as required Minerals, dietary fiber, polyunsaturated fatty acids, other nutrients (coenzyme Q10, carnitine, sesamin, α-riboic acid, inositol, D-kaileunositol, pinitol, phosphatidylserine, phosphatidyl DHA, phosphatidylinositol, taurine, glucosamine, Chondroitin sulfate, S-adnosylmethionine, etc.), stabilizers such as dispersants, emulsifiers, sweeteners, taste ingredients (citric acid, malic acid, etc.), flavors, royal jelly, propolis, agarics, etc. can be blended. . Also, herbs such as peppermint, bergamot, chamomile, lavender and thyme may be blended. In addition, materials such as theanine, dehydroepiandosterone, and melatonin can be blended.
When the clathrate of the present invention is used as a food or a supplement, the usage and dosage are not particularly limited, but the usage and dosage described in the section on inhibitors for increasing blood glucose level can be applied.

(3) Cosmetic Product The cosmetic product of the present invention contains the loquat tissue culture extract itself or the inclusion product of the present invention.
Examples of cosmetics containing the inclusion of the present invention include creams, emulsions, lotions, microemulsion essences, bathing agents and the like, and fragrances and the like may be mixed.
When the clathrate of the present invention is used as a cosmetic, an amount of 1 mg to 20 g, preferably 10 mg to 2.0 g, more preferably 100 mg to 200 mg per 100 g of cosmetic can be blended.
Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.

Production of loquat leaf culture extract In this example, an extract was extracted from callus culture of loquat leaf.
Calli culture of loquat leaves was performed according to the description in JP-T-2009-500013.
The culture was dried and washed with 50% ethanol. Next, after extracting with ethanol, it filtered, the solution part was dried with the vacuum dryer, and this was made into the loquat leaf culture extract.
As a result of analyzing the obtained extract by the method shown in Table 1, it was shown that it had the composition shown in Table 1.

Production of inclusions (1) Inclusion 1
100 mg of loquat leaf culture extract obtained in Example 1, 500 mg of 5-fold amount of γ-cyclodextrin, 2 ml of THF-methanol (volume ratio = 20: 1) and 2 ml of ion-exchanged water are mixed and well in a mortar for about 20 minutes. Kneaded to make a paste. The solvent added in the final stage was almost volatilized, and the residue (remaining powder) was dried with a vacuum dryer for 5 hours.

(2) Inclusion thing 2
Biwa leaf culture extract 100 mg, 10 times the amount of β-cyclodextrin 1 g, THF-methanol (volume ratio = 20: 1) 2 ml and ion-exchanged water 2 ml were mixed and kneaded well in a mortar for about 20 minutes to form a paste. . The solvent added in the final stage was almost volatilized, and the residue (remaining powder) was dried with a vacuum dryer for 5 hours.
Table 2 shows the relationship between the clathrate prepared and water solubility.

In water solubility, “◯” is 0.05 mg / ml or more and 0.5 mg / ml or less (dissolves well by stirring or ultrasonic wave), “Δ” is 0.01 mg / ml or more and 0.05 mg / ml or less (stirring or ultrasonic wave or addition) “X” (dissolves by temperature) indicates 0.0 mg / ml (cannot be dissolved by diffusion and ultrasonic or heating). “Control” is loquat leaf culture extract itself which is not included.
As a result of measuring the triterpene content in the inclusion product of loquat leaf culture extract-cyclodextrin by HPLC, it was confirmed that it was dissolved in water at a theoretical weight without being decomposed in the production process.

<Production Example 1>
The loquat leaf culture extract prepared in Example 1 was encapsulated to produce a capsule preparation containing 444 mg of loquat leaf culture extract per 10 capsules.
Table 3 shows the composition per 10 capsules.

Suppresses blood glucose level
Changes in fasting blood glucose levels when eating loquat leaf culture extract were studied in healthy adult men and women aged 20 to 64 years old.
The number of subjects was 22, and 11 healthy adult subjects (test group 1) and 11 subjects with a fasting blood glucose level of 100 to 126 mg / dL (test group 2) were used.
Under the supervision of doctors, the study will give full consideration to the privacy of the subject and the protection of personal information, and will be reviewed and approved by the ethical review board of the medical institution, and the subject shall obtain informed consent. I did it.

The capsules produced in Production Example 1 as test food were given to the subject 10 capsules per day. In 1 capsule, loquat leaf culture extract contains 44.4mg. The implementation period was 4 weeks. The blood glucose level was measured by an ordinary clinical test method (hexokinase method).
The results are shown in FIG.
In healthy subjects (Study Group 1), fasting blood glucose increased 0.4 ± 5.8 mg / dL at the fourth week after ingestion.
In subjects with high blood glucose levels (Test Group 2), fasting blood glucose decreased by 14.5 ± 13.8 mg / dL at the fourth week after ingestion.
It was confirmed that ingestion of loquat leaf culture extract 444 mg / day significantly reduced fasting blood glucose level.
This example showed that fasting blood glucose levels were reduced by food. Such a blood glucose level lowering action is the first example in food.

  The loquat tissue culture extract of the present invention or the inclusion product of the culture extract and cyclodextrin has high intestinal absorption and is useful as a functional food for lowering blood glucose level.

Claims (13)

  An inclusion product of a culture extract of loquat tissue and cyclodextrin or a derivative thereof.   The inclusion according to claim 1, wherein the loquat tissue is loquat leaves.   Consists of corosolic acid, maslinic acid, tormic acid, ursolic acid, 2α, 19α-dihydroxy-3-oxours-12-en-28-oic acid, oleanolic acid and hupenic acid and their pharmaceutically acceptable salts An inclusion product of at least one selected from the group and cyclodextrin or a derivative thereof.   The clathrate according to any one of claims 1 to 3, wherein the water solubility is higher than the water solubility of the non-inclusion loquat leaf culture extract.   The clathrate according to claim 4, wherein the intestinal absorbability is improved as compared with the intestinal absorbability of the non-clathed loquat leaf culture extract by improving water solubility.   6. Inclusion according to claim 5, wherein the intestinal absorbability is improved, so that the amount of components transferred in the clathrate is increased in blood than the amount of components in the non-inclusion loaf leaf culture extract. object.   The clathrate according to claim 6, which exhibits an effect of suppressing an increase in blood sugar level when components in the clathrate are transferred into the blood.   The blood sugar level rise inhibitor containing the culture extract of loquat tissue, or the clathrate of any one of Claims 1-7.   The inhibitor according to claim 8, wherein the loquat tissue is loquat leaves.   The inhibitor according to claim 8 or 9, wherein the blood glucose level is a fasting blood glucose level.    The inhibitor according to claim 10, which is used for a human having a fasting blood glucose level of at least 100 mg / dl.   A food containing a culture extract of loquat tissue or an inclusion product according to any one of claims 1 to 7. Cosmetics containing the culture extract of loquat tissue, or the inclusion of any one of Claims 1-7.