JP5405067B2 - Antioxidant composition containing bee pup - Google Patents

Description

  The present invention relates to an antioxidant containing bees as an active ingredient, an antioxidant containing bees and propolis, an antioxidant containing bees, propolis and royal jelly, and an oral intake containing the antioxidant. Relates to the composition.

  In general, oxygen is essential for living organisms, but on the other hand, it is also known to cause harm to living organisms. In particular, it has been pointed out that peroxides generated by oxidation of active oxygen such as hydroxy radicals and lipids cause serious damage to living bodies. In recent years, it is known that oxidative stress, particularly vascular endothelial cell oxidative stress is also involved in the development of thrombosis and cerebral circulation-related diseases which are positioned as lifestyle diseases. Furthermore, it is known that oxidative stress is involved in aging promotion and induces skin wrinkles, spots, dullness and the like. For this reason, controlling a harmful oxidation reaction in a living body is very useful for health maintenance and beauty, and various antioxidants are attracting attention.

  In addition, it is known that oxidation is involved in quality deterioration of compositions for oral intake such as food. In particular, the oxidation of fats and oils causes deterioration in color and flavor and nutritional value, and in some cases, peroxides generated by oxidation may cause digestive disorders. As an antioxidant for preventing oxidation of the composition for oral intake, ascorbic acid, tocopherol, catechin, dibutylhydroxytoluene and the like are used.

  Traditionally, in some parts of Japan, bee larvae or moths are used for food as “bees”. It has been reported that a bee is a food having a specific amino acid composition and has a fatigue recovery effect (Patent Documents 1 to 3).

  It is known that propolis, which is a sticky substance used as a material for beehives, contains various sap activity including antioxidants as a component, including sap of plants collected by bees from the field (Patent Documents 4 to 4). 7). For this reason, propolis is used as a raw material for health foods.

Royal jelly is a milky white creamy substance secreted from the secretory gland (pharyngeal gland) of the worker's head, and is a substance that feeds the bee queen. Royal jelly is often used in health foods for the purpose of nutrition and tonic, and is also used as a component of skin external preparations such as cosmetics and quasi drugs (Patent Documents 8 to 11).
JP 2002-17296 A JP 2006-197911 A JP 2001-204421 A JP 2000-44479 A JP 2002-235084 A JP 2002-306092 A Japanese Patent Laid-Open No. 2005-220087 JP 2001-292712 A JP 2002-20226 A JP 2003-211981 A JP 2006-280249 A

  In order to protect the body from oxidative stress and to prevent the oxidation of oral compositions, there is a need for antioxidants that can be taken on a daily basis and that are safe. An object of the present invention is to provide an antioxidant that can be safely ingested. A further object of the present invention is to provide an antioxidant that synergistically exerts an antioxidant effect by containing specific raw materials in combination.

The present inventor has conducted extensive studies to solve the above problems, and has found a good antioxidant effect and anti-aging effect in a composition containing a bee, and has completed the present invention.
According to one aspect of the invention, an antioxidant comprising a bee pup is provided.

  The bee kid used in the present invention is not particularly limited as long as it is a bee larva or moth used for food. For example, honey bees (for example, Apis melifera), honey bees ( Apis cerana), honey bee (Apis koschevnikovi), honey bee (Apis nurenis), honey bee (Apis doras) Apis andreniformis)); hornets (eg, Vespa mandaria japonica, Himesuzu) Bigeye (Vespa ducalis), Kiirosuzumebachi (Vespa simillima xanthoptera), Kogatasuzumebachi (Vespa analis), Mont hornet (Vespa crabro), Tenebrio hornet (Vespa dybowskii), black hornet (Vespula flaviceps), Kio Biho tailed wasp (Dolichovespula media), sharks Hornets (Provespa spp.); And wasps of the waspidae family (e.g., Polists jadwigae, Polists rothneyi), puffer bees (Polistes chinensis), nell ni), it can be used larvae or pupae, such Kiboshi paper wasp (Polistes mandarinus)). In the present invention, for example, honey bees (especially honey bees), giant hornets, killer bees or cross bees, preferably bees of honey bees can be used.

  For example, a queen bee, a worker bee and a male bee, preferably a male bee larva or a moth can be used as the bee cub, and since it has an excellent antioxidant effect, for example, 10 to 24 days after hatching, preferably 20 to 22 Can be used after a day.

  The collected bee kid can be used, for example, by drying, pulverizing, and powdering. For example, the bee cub powder disclosed in Japanese Patent Application Laid-Open No. 2002-17296 (paragraph number [0007]) is used. You may use for this invention. In one embodiment of the present invention, bees frozen at −80 to −5 ° C. after collection are naturally thawed before use, mixed with a stirrer, and the resulting mixture is boiled and sterilized for 10 to 20 minutes, and then frozen. The powder obtained by sieving (10-200 mesh) after drying can be used.

  In the present invention, a bee extract obtained from the bee powder using, for example, a lower alcohol (eg, methanol, ethanol, propanol, isopropanol, butanol, etc.) and / or water as an extraction solvent. Can be used.

In the present invention, an enzyme degradation product obtained by degrading the bee powder with an enzyme such as protease, cellulase or amylase can be used.
In one embodiment of the present invention, water is added to a bee, heated and extracted at 25 to 50 ° C. for 30 minutes to 3 hours with stirring, cooled to room temperature, and then filtered (bee extract). A liquid obtained by optionally adding an excipient to the liquid, followed by spray drying, granulating and sieving can be used as the bees of the present invention.

  In the present invention, handling in production can be facilitated by adding an excipient to the bee extract. Examples of excipients that can be used in the present invention include excipients that can be used in medicine or food, such as dextrin, corn starch, rice flour, lactose, potato starch, reduced starch saccharified product, and silicon dioxide.

  The amount of bees blended in the antioxidant of the present invention is not particularly limited, but is preferably 50% by weight or more, more preferably 60% by weight or more, still more preferably 70% by weight or more, and particularly preferably 80% by weight. It is preferable to contain above.

According to a further aspect of the invention, an antioxidant comprising bees and propolis is provided.
The propolis used in the present invention is not particularly limited as long as it is a propolis commonly used in foods. From the bees of the honey bee family, for example, honeybee and its subspecies (Apis melifera, Apis mellifera adanoni, Apis mellifera scutella). Harvested propolis can be used. Propolis origin plants are not particularly limited, and for example, propolis originating from plants such as bacchus, poplar, hawthorn and eucalyptus can be used.

  As the propolis, for example, a powdered propolis bulk or a propolis extract obtained by a suitable extraction method from the propolis bulk can be used. For example, a lower alcohol (for example, methanol, ethanol, propanol, Propolis extracts obtained using isopropanol, butanol, etc.) and / or water as the extraction solvent can be used. For example, a propolis extract with ethanol and a propolis extract obtained by the method described in JP-A No. 2002-306092 can be used in the present invention. As the propolis extract, for example, those commercially available from Morikawa Kendo Co., Ltd., Al Techno Industry Co., Ltd., and Nissin Honey Co., Ltd. can be used in the present invention.

  Moreover, in this invention, the propolis enzyme degradation product obtained by decomposing | disassembling the said propolis extract with enzymes, such as protease, a cellulase, and an amylase, can be used, for example.

  In one embodiment of the present invention, ethanol is added to the propolis bulk, heated at 30 to 70 ° C. for 1 to 3 hours with stirring, cooled to room temperature, and then the solution obtained by filtration is concentrated to a waxy form. A solid was obtained. The powder obtained by adding an excipient | filler arbitrarily to the said solid substance (propolis extract), spray-drying after that, and granulating and sieving can be used as the propolis of this invention.

  In the present invention, handling in production can be facilitated by adding an excipient to the propolis extract. Examples of excipients that can be used in the present invention include excipients that can be used in medicine or food, such as dextrin, corn starch, rice flour, lactose, potato starch, reduced starch saccharified product, and silicon dioxide.

  The blending ratio of the antioxidant including the bee and propolis of the present invention is not particularly limited, but the bee is preferably 24 parts by weight or more with respect to 1 part by weight of propolis, and 40 parts by weight or more. More preferably, it is more preferably 60 parts by weight or more. Moreover, it is preferable that it is 240 weight part or less with respect to 1 weight part of propolis, and it is more preferable that it is 120 weight part or less. If it is this range, it can be set as the antioxidant which shows the outstanding antioxidant effect.

  The blending amount of bees and propolis in the antioxidant of the present invention is not particularly limited, but is preferably 50% by weight or more in total, more preferably 60% by weight or more, further preferably 70% by weight in total. As described above, the total content is preferably 80% by weight or more.

In a further aspect of the invention, an antioxidant is provided comprising bees, propolis and royal jelly.
The royal jelly used in the present invention is not particularly limited as long as it is a royal jelly provided for edible use. For example, it is described in “Fair Competition Rules for the Labeling of Royal Jelly” (prepared by the National Royal Jelly Fair Trade Council). "Raw royal jelly", "dried royal jelly" and "prepared royal jelly" can be used, both of which include, for example, honey bees (especially the bees and their subspecies (Apis melifera, Apis mellifera adanisonii, Royal jelly collected from Apis melifera scutellata)), preferably royal jelly collected from honey bees can be used. In the present invention, for example, it is preferable to use a powder obtained by drying royal jelly collected from honey bees.

  In one embodiment of the present invention, a powder obtained by pulverizing a stock solution of royal jelly frozen at −80 to −5 ° C. after collection with a stirrer after freeze-drying and sieving (10 to 200 mesh) is obtained according to the present invention. Can be used as a royal jelly.

  In the present invention, a royal jelly extract obtained from the royal jelly powder using, for example, a lower alcohol (for example, methanol, ethanol, propanol, isopropanol, butanol, etc.) and / or water as an extraction solvent can be used.

  In the present invention, an enzyme degradation product obtained by degrading the royal jelly powder and the royal jelly extract with an enzyme such as protease, cellulase, or amylase can be used.

  In one embodiment of the present invention, water is added to royal jelly, heated and extracted at 25 to 50 ° C. for 30 minutes to 3 hours with stirring, cooled to room temperature, and filtered to obtain a solution (royal jelly extract). The powder obtained by adding an excipient, followed by spray drying, granulating and sieving can be used as the royal jelly of the present invention.

  In the present invention, handling in production can be facilitated by adding an excipient to the royal jelly extract. Examples of excipients that can be used in the present invention include excipients that can be used in medicine or food, such as dextrin, corn starch, rice flour, lactose, potato starch, reduced starch saccharified product, and silicon dioxide.

  The blending ratio of the antioxidant including the bee, propolis and royal jelly of the present invention is not particularly limited, but the bee is preferably 6 parts by weight or more with respect to 1 part by weight of propolis, and 12 parts by weight. More preferably, it is more preferably 16 parts by weight or more, and particularly preferably 24 parts by weight or more. Moreover, it is preferable that it is 240 weight part or less with respect to 1 weight part of propolis, and it is more preferable that it is 120 weight part or less. Furthermore, the amount of bees is preferably 0.3 parts by weight or more with respect to 1 part by weight of royal jelly, more preferably 0.6 parts by weight or more, and further preferably 0.86 parts by weight or more. . Moreover, it is preferable that it is 60 weight part or less with respect to 1 weight part of royal jelly, and it is more preferable that it is 2 weight part or less. If it exists in this range, it can be set as the antioxidant which shows the outstanding antioxidant effect.

  The blending amount of bees, propolis and royal jelly in the antioxidant of the present invention is not particularly limited, but is preferably 50% by weight or more in total, more preferably 60% by weight or more, and further preferably 70% in total. It is preferable to contain 80% by weight or more, particularly preferably 80% by weight or more.

Each raw material used in the present invention contains components specific to each raw material.
As a component peculiar to a bee child, chitin which is an outer shell component of a moth or an adult is mentioned, and 1 to 10 weight% is contained in the dried powder of a bee child.

As a component peculiar to propolis, p-coumaric acid is mentioned, and 0.5 to 5 weight% is contained in the powder of 50 weight% of propolis extract.
As a component peculiar to royal jelly, decenoic acid is mentioned, and 2 to 10% by weight is contained in the royal jelly dry powder.

  In one embodiment of the present invention, the amount of chitin derived from bees contained in the antioxidant of the present invention is, for example, 0.0005% by weight or more, particularly 0.001% by weight or more, for example, 0.1% by weight. % Or less, particularly 0.05% by weight or less. The amount of p-coumaric acid derived from propolis contained in the antioxidant of the present invention is, for example, 0.00001% by weight or more, particularly 0.00004% by weight or more, for example, 0.005% by weight or less, particularly 0 0.002% by weight or less. Furthermore, the amount of decenoic acid derived from royal jelly contained in the antioxidant of the present invention is, for example, 0.0015% by weight or more, particularly 0.003% by weight or more, for example 0.3% by weight or less, particularly 0.08%. % By weight or less.

In addition, each component in the antioxidant of this invention can be measured by a method well-known to those skilled in the art, respectively, and a compounding quantity can be computed based on the measured value.
Since the antioxidant of the present invention exhibits an excellent antioxidant effect, it can be used as it is or in a composition for oral consumption. For example, when the antioxidant of the present invention is used in a composition for oral consumption, it is not only used for treatment, improvement and / or prevention of various disorders caused by active oxygen, but also for oral intake. It can also be used effectively as an antioxidant for the composition itself. The compounding amount of the antioxidant in the composition for oral intake of the present invention is not particularly limited, but in order to obtain an excellent antioxidant effect, the ascorbic acid equivalent is preferably 0.0003% by weight or more. Here, the composition for oral intake includes, for example, a food composition, a pharmaceutical composition, and the like.

The food composition of the present invention can be a health food (such as a nutritional functional food, a food for specified health use, or a supplement) having an antioxidant action, a food for the sick, and the like.
The food composition of the present invention contains the antioxidant of the present invention (preferably 0.0003 wt% or more as an ascorbic acid equivalent), and is excellent even without a known antioxidant such as ascorbic acid or tocopherol. It has an antioxidant effect, is safe, and has excellent long-term stability in terms of color, flavor and nutritional value. Moreover, since the food composition of the present invention has high antioxidant activity, various disorders caused by active oxygen present in the living body (myocardial infarction, arteriosclerosis, diabetes, cancer) due to the ingestion of the food composition , Stroke, cataracts, stiff shoulders, coldness, hypertension, senile dementia, spots, buckwheat, wrinkles, melasma, dullness, other skin deterioration, etc.), and skin firmness, skin gloss It can be ingested for cosmetic paste, skin roughness, skin texture, skin dullness, skin breakout / acne, skin elasticity improvement and / or prevention.

  The pharmaceutical composition of the present invention contains the antioxidant of the present invention (preferably 0.0003% by weight or more as an ascorbic acid equivalent) and has a very excellent antioxidant activity. , Various disorders caused by active oxygen present in the body (myocardial infarction, arteriosclerosis, diabetes, cancer, stroke, cataract, stiff shoulder, coldness, hypertension, senile dementia, spots, freckles, wrinkles, melasma, For treatment, improvement and / or prevention of skin dullness, other skin weakness, etc., as well as skin firmness, skin gloss, makeup paste, skin roughness, skin texture, skin dullness, skin pimples / acne, It can be taken for the treatment, improvement and / or prevention of skin elasticity. Moreover, the pharmaceutical composition of the present invention itself is hardly oxidized, and can prevent oxidation or discoloration of active ingredients other than the antioxidant of the present invention for a long period of time.

The composition for oral intake of the present invention can be blended with components such as conventionally known colorants, preservatives, fragrances, flavors, coating agents and the like as long as they do not impair the effects of the present invention.
Moreover, the composition for oral intake of this invention can mix | blend the processed material of a plant, a functional component, or a medicinal component in addition to the said antioxidant in the range which does not impair the effect of this invention. In addition, these components may be used alone or in combination with the above antioxidant, or two or more kinds may be arbitrarily combined and used in combination with the above antioxidant.

  Examples of plants that can be used in combination are plant extracts having an antioxidant action and the like (e.g., Akamegashi, Oyster, Mulberry, Hypericum perforatum, Rhizoma, Sakuyo garage, Yaba Santa, Rosehip, Extracts such as Asunaro, Azalea, Itadori, Ichiakuso, Apricot, Keikanka, Hakukayumatou, Birch, Hawthorn, Achillea millefolium, Tarayou, Dokudami, Tormentilla, Bakumondo, Hiba, Grape, Mukuroji, Mokka, Ganoderma, Roman cuttlefish Plant extracts having an action (for example, grape seed, hijiki, moroheiya, hamasge, cereals (for example, beans, rice and wheat), herbs (for example, myrrh, basil, thyme, etc.), cacao extract, coffee sauce Vinegar, plum meat extract, extracts such as hawthorn extract, and chlorella); soybean extract having a female hormone-like action and black cohosh extract; such as blood pressure-lowering effect and rice germ extract having a mental stability can be mentioned.

  Examples of ingredients that improve the nutritional aspect of the composition for oral consumption of the present invention include phytic acid, black garlic, black vinegar, royal jelly, black pepper, sorghum, tonkat ant, paffia, muirapuama, tahibo, taichi Examples include carrots, red scenic heavens, rahan fruit, litchi, bitter gourd, caiapo, yacon, yerba mate, sugina and reindeer horns.

Examples of functional components include antioxidants, hypoglycemic agents, anticholesterol agents, and immunostimulants.
Here, examples of antioxidants include, but are not limited to, dry yeast, glutathione, lipoic acid, quercetin, catechin, coenzyme Q10, enzodinol, proanthocyanidins, anthocyanidins, anthocyanins, carotenes, lycopene, flavonoids, resaveratrol , Isoflavones, zinc, ginkgo biloba, moon peach leaf, hibiscus, and melatonin.

  Examples of the hypoglycemic agent include, but are not limited to, indigestible dextrin, guava leaf, wheat albumin, L-arabinose, bean extract, mulberry leaf, ginger, salacia, α-linolenic acid, amacha, barley, kidachi aloe, dandelion , Daidai, Korean thistle, garlic, pearl barley, banaba, bilberry, black cohosh, makomo, kotarahim, and tanaka.

  Examples of anticholesterol agents include, but are not limited to, soy protein, phospholipid-bound soy peptide, chitosan, plant sterol ester, plant sterol, plant stanol ester, resistant digestive dextrin, sodium alginate, psyllium seed coat, astaxanthin, inositol, coenzyme A, Calcium, Magnesium, Carnitine, Silk protein, Taurine, Methionine, α-Linolenic acid, Guam gum, Chondroitin sulfate, Achachaul, Alfalfa, Ginkgo, Psyllium, Barley, Oats, Olives, Gadju, Gymnema, Cats claw, Cuco, Chlorella Spirulina, hawthorn, soy saponin, chili, garlic, bilberry, safflower, yucca, ruffa, agaricus, red yeast rice.

  Examples of immunostimulants include agaricus, lactoferrin, cordyceps, arginine, tryptophan, valine, leucine, chitin, chitosan, aloe, kidachi aloe, echinacea, ogi, cat's claw, wolfberry, spirulina, pearl barley, safflower, maca, makomo, rafuma Can be mentioned.

  Examples of medicinal ingredients include vitamins, amino acids, peptides, proteins, minerals (iron, zinc, magnesium, iodine, etc.) and fatty acids (EPA, DHA, etc.).

  Examples of vitamins include vitamins belonging to the vitamin A group (for example, retinal, retinol, retinoic acid, carotene, dehydroretinal, lycopene and pharmacologically acceptable salts thereof (for example, retinol acetate, retinol palmitate). Etc.), vitamins belonging to the vitamin B group (for example, thiamine, thiamine disulfide, dicetiamine, octothiamine, chicotiamine, bisibtiamine, bisbenchamine, prosultiamine, benfotiamine, fursultiamine, riboflavin, flavin adenine Dinucleotide, pyridoxine, pyridoxal, hydroxocobalamin, cyanocobalamin, methylcobalamin, deoxyadenocobalamin, folic acid, tetrahydrofolate, dihydrofolate, nicotinic acid, nicotinic acid Nicotinic alcohol, pantothenic acid, panthenol, biotin, choline, inositol, pangamic acid and their pharmacologically acceptable salts thereof (eg thiamine hydrochloride, thiamine nitrate, dicetiamine hydrochloride, fursultiamine hydrochloride, Riboflavin butyrate, flavin adenine dinucleotide sodium, pyridoxine hydrochloride, pyridoxal phosphate, calcium pyridoxal phosphate, hydroxocobalamin hydrochloride, hydroxocobalamin acetate, calcium pantothenate, sodium pantothenate, etc.), vitamins belonging to the vitamin C group (ascorbic acid And derivatives thereof, erythorbic acid and derivatives thereof and pharmacologically acceptable salts thereof (for example, sodium ascorbate, sodium erythorbate, etc.), Vitamins belonging to the group Tamine D (for example, ergocalciferol, cholecalciferol, hydroxycholecalciferol, dihydroxycholecalciferol, dihydrotachysterol and pharmacologically acceptable salts thereof), vitamins belonging to the vitamin E group [For example, tocopherol and its derivatives, ubiquinone derivatives and pharmacologically acceptable salts thereof (such as tocopherol acetate, tocopherol nicotinate, tocopherol succinate, tocopherol calcium succinate, etc.), other vitamins (eg, carnitine, Ferulic acid, γ-oryzanol, orotic acid, rutin (vitamin P), eriocitrin, hesperidin and pharmacologically acceptable salts thereof (such as carnitine chloride).

  Examples of amino acids include leucine, isoleucine, valine, methionine, threonine, alanine, phenylalanine, tryptophan, lysine, glycine, asparagine, aspartic acid, serine, glutamine, glutamic acid, proline, tyrosine, cysteine, histidine, ornithine, hydroxyproline. , Hydroxylysine, glycylglycine, aminoethylsulfonic acid (taurine), cystine or pharmacologically acceptable salts thereof (for example, potassium aspartate, magnesium aspartate, cysteine hydrochloride, etc.). Preferred examples include branched chain amino acids such as valine, leucine and isoleucine, glutathione, cysteine, glutamic acid, glycine, serine, tryptophan, tyrosine, phenylalanine, histidine, methionine, threonine, lysine, cystine, arginine, alanine, aspartic acid, proline, Aminoethylsulfonic acid.

  In addition, in the composition for oral intake of the present invention, the above-mentioned antioxidant may be used as it is as a composition for oral intake, and for example, a granule (including dry syrup) so as to facilitate continuous intake. , Various solid preparations such as capsules (soft capsules, hard capsules), tablets (including chewables), powders (powder), pills, or liquids for internal use (liquids, suspensions, syrups) It is desirable to prepare it in the form of a liquid preparation such as a liquid preparation. Among them, capsules and tablets are preferable from the viewpoint of stability of each component and ease of ingestion, but are not particularly limited. .

  The composition for oral consumption of the present invention in the form of capsules and tablets can use a known pharmaceutically acceptable carrier, and is a usual formulation method adopted in the fields of medicine and food (especially supplements). Can be applied. For example, a tablet can be prepared by adding and blending each component according to the formulation, crushing, granulating, drying, sizing and mixing, and tableting the resulting preparation mixture.

  Furthermore, additives for formulation, for example, excipients, lubricants, binders, disintegrating agents, fluidizing agents, dispersing agents, wetting agents, preservatives, thickeners, pH adjustments as necessary An agent, a coloring agent, a flavoring agent, a surfactant, a solubilizing agent and the like can be blended. Moreover, when making it into the form of a liquid agent, thickeners, such as pectin, xanthan gum, and guar gum, can be mix | blended. Thereby, even if it is a case where many components which are hard to melt | dissolve in water, such as a bee, it can disperse | distribute successfully in a liquid agent, and stability can also be improved. Moreover, it can also be set as a coating tablet using a coating agent, or it can also be set as a paste-form glue. Furthermore, even if it is a case where it prepares in another form, what is necessary is just to follow the conventional method.

  Furthermore, the composition for oral consumption of the present invention in the form of granules, powders, liquids, and the like includes, for example, various foods and beverages such as beverages, confectionery, breads and soups; various pet foods such as dog foods and cat foods, etc. In addition, it is prepared as various foods and drinks. The method for producing these foods and drinks is not particularly limited as long as the effects of the present invention are not impaired, and may follow the methods used by those skilled in the art for each application.

  The dose of the composition for oral intake of the present invention can be appropriately selected depending on the body shape, age, physical condition, severity, etc. of the subject. For example, the antioxidant of the present invention based on an adult weighing 60 kg. Is used at a dose of 100 to 3000 mg / day, preferably 300 to 2000 mg / day, more preferably 500 to 1000 mg / day. The composition for oral ingestion may be administered in a single dose or multiple doses, and may be used in combination with other drugs such as other antioxidants.

  In still another aspect of the present invention, there is provided a method for enhancing antioxidation of propolis, characterized by adding bees to propolis, and a method for enhancing antioxidation of propolis, comprising adding bees and royal jelly to propolis To do.

  The above-mentioned bees, propolis and royal jelly that can be used in the method for enhancing antioxidation of propolis of the present invention can be used. In the method for enhancing antioxidation of propolis of the present invention, the mixing ratio of propolis and bee and the mixing ratio of propolis and royal jelly are not particularly limited, but are preferably as described above.

  The antioxidant of the present invention has an excellent antioxidant action. As a result, (1) discoloration, deterioration of flavor, and nutritional value of the composition for oral intake containing the antioxidant are prevented, and the effect of the active ingredient contained in the composition for oral intake is prolonged. (2) Various disorders caused by active oxygen present in the living body (myocardial infarction, arteriosclerosis, diabetes, Cancer, stroke, cataract, stiff shoulders, coldness, hypertension, senile dementia, spots, buckwheat, wrinkles, melasma, dullness, other skin deterioration, etc.), treatment and improvement and / or prevention, and skin firmness, skin It is possible to improve and / or prevent skin paste, makeup paste, skin roughness, skin texture, skin dullness, skin breakout / acne, skin elasticity and the like.

  EXAMPLES Hereinafter, although the preferable Example of this invention is described in detail, this invention is not limited to these Examples. In addition, the percentage shown in this specification means weight% unless there is particular mention.

The bees, propolis, and royal jelly used for the preparation test of each sample were prepared by the following method.
[Bees]
20 to 22 days after hatching of honeybees were collected and frozen at -18 ° C. Thereafter, the bees that were naturally thawed before use were mixed with a stirrer, and the resulting mixture was sterilized by boiling for 60 minutes. Thereafter, it was freeze-dried and sieved (50 mesh) to obtain a bee powder. The bee powder was used in the following tests ("Antioxidant activity measurement test", "Efficacy confirmation test in humans", and "Mouse skin layer thickness measurement test").

  In addition, the chitin content of the obtained bee powder was measured in accordance with “Chapter 5 Analysis of chitin and chitosan” in the Chitin / chitosan experiment manual (Technical Publishing, edited by the chitin / chitosan study group). It was 0% by weight (per dry powder weight).

[Propolis]
Ethanol was added to the propolis bulk collected from the honeycomb and heated at 50 ° C. with stirring for 2 hours. After cooling to room temperature, the solution obtained by filtration was concentrated to obtain a waxy solid (propolis extraction). object). A dextrin having the same weight as the solid was added to the obtained solid to obtain a propolis powder. The propolis powder was used in the following tests.

  Further, the obtained propolis powder was subjected to a high performance liquid chromatography method (see High Performance Liquid Chromatography Handbook (Revised 2nd edition), edited by the Japan Society for Analytical Chemistry, Kanto Chapter), and the content of p-coumaric acid was measured. The content of coumaric acid was 2.1% by weight (per 50% powder weight of propolis extract).

  Moreover, the amount of propolis in Tables 2 to 6 in the following “antioxidant activity measurement test” indicates the amount of propolis extract contained in the propolis powder (propolis extract content: 50% by weight). It has been confirmed that the dextrin used as an excipient has no antioxidant activity.

[Royal Jelly]
The solid obtained by freeze-drying the royal jelly stock solution frozen at −18 ° C. after collection was pulverized with a pulverizer, and the powder obtained by sieving (50 mesh) was used as the royal jelly. The resulting royal jelly powder was subjected to a high performance liquid chromatography method (see High Performance Liquid Chromatography Handbook (Revised 2nd edition), edited by the Japan Society for Analytical Chemistry, Kanto Branch), and the content of decenoic acid was measured. The rate was 5.9% by weight (per dry powder weight).

[Test Example 1] Antioxidant activity measurement test [Preparation of sample solution]
In accordance with Tables 1 to 6 below, 100 μL is taken from the powdered dimethyl sulfoxide solution of each sample prepared to have a predetermined concentration, and after centrifugation (5000 rpm × 5 minutes), 10 μL is taken from the supernatant to obtain the sample solution. did.

[Measuring method]
Diphenyl-2-picrylhydrazyl (hereinafter referred to as DPPH; Nacalai Tesque, # 13933-61) was dissolved in ethanol (Wako, special grade) to prepare a 200 μM DPPH / ethanol solution. A DPPH solution (190 μL) and a sample solution (10 μL) are added to each well of a 96-well microplate, stirred for 5 minutes, and allowed to stand at room temperature for 30 minutes. Thereafter, a microplate reader (manufactured by Tecan Japan, product name: The absorbance at a wavelength of 530 to 550 nm was measured with GENios.

[Create calibration curve]
First, changes in concentration and absorbance were measured using L-ascorbic acid as a sample solution. The results are shown in Table 1. In the table, Asc represents a concentration (% by weight) of L-ascorbic acid, Abs represents a measured value of absorbance at 550 nm of the sample, and (Bla-Abs (550 nm)) represents (blank absorbance measured value−sample absorbance measured value). In addition, a calibration curve was prepared from Table 1, the following formula was obtained, and the antioxidant activity of each sample was shown as L-ascorbic acid equivalent (Asc (%) [EQ]).
(Blank absorbance measurement value−Sample absorbance measurement value) = 24.566 × (L-ascorbic acid concentration) +0.0029

Antioxidant activity of bee powder Antioxidant activity was measured using bee powder as a sample solution, and L-ascorbic acid equivalent was calculated. The results are shown in Table 2.

From the above results, it was confirmed that the bees had antioxidant activity.
Here, ascorbic acid generally used as an antioxidant for foods and beverages and cosmetics and its raw materials, for example, the amount used when injected or blended into a food material as an antioxidant is 0.0003 to About 0.0005% by weight (see Food Additives Official Document (8th Edition), Akio Tanimura, Yodogawa Shoten, 2007). As a result of this test, an antioxidant activity equivalent to 0.0003% by weight of L-ascorbic acid was obtained at 0.05% by weight of the bee, so that the bee is excellent as an antioxidant. I understood.

Antioxidant activity measurement test of propolis and royal jelly Antioxidant activity was measured using each of propolis powder and royal jelly powder as a sample solution, and L-ascorbic acid equivalent was calculated. The results are shown in Table 3.

Propolis has been confirmed to have an antioxidant effect, but royal jelly has a very low antioxidant effect, which is insufficient for use as a single antioxidant.
Antioxidant activity measurement test of a mixture of bee and propolis Antioxidant activity was measured as a sample solution of a mixture of bee powder and propolis powder. The results are shown in Table 4. The theoretical values in the table are the sum of the Asc (%) [EQ] measurements (Tables 2 and 3) when bee and propolis are used alone at each concentration, and the synergistic effect is the Asc of the mixture. (%) [EQ] Ratio of measured value to theoretical value.

  In Table 4, in the mixture containing 24 to 240 parts by weight of bees with respect to 1 part by weight of propolis, the antioxidant effect exceeding the theoretical value was confirmed. From this result, it was confirmed that the antioxidant activity is synergistically increased by mixing the bee and the propolis.

Antioxidant activity measurement test of a mixture of bee, propolis, and royal jelly Antioxidant activity of a mixture of bee powder, propolis powder, and royal jelly powder as a sample solution was measured in Table 5 below. The results are shown in Table 5. The theoretical values in the table are the sum of Asc (%) [EQ] measurements (Tables 2 and 3) when bee, propolis, and royal jelly are used alone at each concentration, and the synergistic effect is It is the ratio of the Asc (%) [EQ] measured value of the mixture to the theoretical value.

  In Table 5, in the mixture containing bee, propolis, and royal jelly, when the bee child is included in an amount of 6-240 parts by weight with respect to 1 part by weight of propolis, or 0.0. In the case of containing 3 to 60 parts by weight, the antioxidant effect exceeding the theoretical value was confirmed. Furthermore, comparing the results of Table 5 with the results of Table 4, it can be seen that mixing three kinds of bees, propolis, and royal jelly provides higher antioxidant activity than the two mixtures.

[Test Example 2] Efficacy confirmation test in humans Antioxidants treat, ameliorate and / or prevent various disorders caused by oxidation in animals receiving such antioxidants. Therefore, in order to confirm the effectiveness of the composition for oral intake of the present invention, an intake test using a human monitor was conducted to examine the effect on the skin.

[Preparation of tablets]
Tablets with the formulations shown in Table 6 were prepared according to conventional methods. Specifically, each component was blended according to the formulation, pulverized, granulated, dried, sized and mixed, and then tableted by a conventional method to form a tablet. As the bee powder, propolis powder, and royal jelly powder, those prepared as described above were used. The blending ratio of each component is 65.6 parts by weight of the honey powder with respect to 1 part by weight of the propolis extract, and 1.3 parts by weight of the honey powder with respect to 1 part by weight of the royal jelly powder.

Evaluation of skin by questionnaire Questionnaire about subjective facial skin condition at the start of ingestion and 1 month after ingestion to 19 female monitors aged 20 to 40 years who took 5 tablets a day for 1 month. Went. The results are shown in Table 7.

  From the results in Table 7, all items of skin firmness, skin shine, makeup paste, skin roughness, skin texture, skin dullness, skin pimples and acne are observed at the beginning of intake compared to the start of intake. The tendency for the evaluation to increase was observed.

Measurement of changes in skin elasticity and water content In order to measure objective changes in skin condition, 11 monitors were tested for facial skin (cheekbone) at the start of administration and at 1 month after the start of administration. Elasticity and moisture content were measured.

[Measuring method]
The measurement was carried out after washing for 20 minutes in a room adjusted to a humidity of 60% and a temperature of 25 ° C., and the elasticity value was measured using a Cutometer MPA580 (Courage + Khazaka electronic) and the Uf value was measured according to the attached manual. . The higher the skin elasticity, the higher the Uf value. The moisture value was measured as electrical conductivity (μS) according to the attached manual using Skicon200EX (manufactured by IBIS Corporation). The higher the moisture content on the skin surface, the higher the electrical conductivity.

  The results are shown in FIG. 1 and FIG. It was confirmed that the skin elasticity was significantly improved at the first month of ingestion compared to the start of ingestion (p <0.05). As for the moisture value, no significant difference was confirmed, but an increasing tendency was confirmed.

From the above test, it was revealed that the composition for oral intake of the present invention has a cosmetic effect.
[Test Example 3] Mouse skin layer thickness measurement test In order to examine the effect of the antioxidant of the present invention on the skin tissue in more detail, the following feeding test was performed using mice.

[Sample preparation]
Samples shown in Table 8 below were prepared. As the bee powder, the royal jelly powder and the propolis powder, those prepared as described above were mixed at a weight ratio of 6: 5: 0.3 (0.15 as the propolis extract). Vitamin Mix and Mineral Mix were used for CLEA purification by CLEA Japan. The milk casein used contained 86.2% protein by weight, corn starch contained 86.3% by weight carbohydrate, corn oil contained 100% by weight of fat, and feeds 1 to 3 all had a protein content of 20% by weight. It was prepared as follows.

[Test method]
Hairless mice (Japan LSC Co., Ltd .: BALB / c Slc-nu / nu (SPF) 3 weeks old) were divided into groups A to E (each group n = 6). Feeded according to the feeding schedule. Specifically, each group was given a low protein diet (containing 6% protein) for 3 weeks to acclimatize, then group A received feed 1, group B fed 2 and group C fed 3 The D group continued to receive a low protein diet. Group E was sacrificed after 3 weeks as a control group and the skin layer thickness was measured.

  Seven weeks after the start of the test, the mice in groups A to D were sacrificed, and the skin layer thickness (dermis layer thickness) was measured. Specifically, cut about 1 cm square of the skin on the right side from the spinal line at the center of the back, fix it with formalin, prepare an Elastica-Wangyson (EVG) -stained specimen, and make a micrograph of the specimen (Figure 3). I took a picture. The thickness of the dermis layer was determined by measuring the distance at any 10 locations in the dermis layer portion stained dark purple on the photograph and dividing the average value by the magnification of the microscope.

  The measurement results are shown in FIG. The thickness of the dermis layer of group A mice fed diet 1 containing a mixture of bee powder, royal jelly powder and propolis powder compared to group B and group C mice fed a diet containing the same amount of protein. Significantly increased (FIGS. 3 and 4).

[Prescription example]
Formulation Examples 1 to 26 (tablets)
Compositions having the formulations shown in Table 10 (Prescription Examples 1 to 26) were prepared as tablets according to a conventional method. Specifically, each component was blended according to the formulation, pulverized, granulated, dried, sized and mixed, and tableted according to a conventional method to prepare a tablet.

Formulation Examples 27-41 (granule)
Compositions (formulation examples 27 to 41) having the formulations described in Table 11 were prepared as granules according to a conventional method. Specifically, each component was blended according to the formulation, and mixed, granulated, dried and sized according to a conventional method to form a granule.

Formulation Examples 42-53 (Liquid)
Compositions (formulation examples 42 to 53) having the formulations described in Table 12 were prepared as liquids according to conventional methods. Specifically, each component was blended according to the formulation and mixed according to a conventional method to form a liquid preparation.

It is a graph which shows the test result regarding the change of the skin elasticity value of the face by administration of the composition of this invention. An asterisk (*) in the figure indicates a significant difference p <0.05. It is a graph which shows the test result regarding the change of the skin skin moisture value by administration of the composition of this invention. It is an example of the microscope picture of the skin tissue cross section of the AE group mouse of Table 12. The scale bar indicates 200 μm. It is a graph which shows the test result regarding the skin layer thickness (dermis layer thickness) of the mouse | mouth which administered the composition of this invention. In the figure, an asterisk (**) indicates a significant difference p <0.01, and (***) indicates a significant difference p <0.001.

Claims (6)

  Antioxidants including bees and propolis.   The antioxidant of Claim 1 containing 6-240 weight part of bees with respect to 1 weight part of propolis.   Furthermore, the antioxidant of Claim 1 or 2 containing a royal jelly. A method for enhancing the antioxidant activity of propolis used as an antioxidant (excluding a method for administration to humans), which comprises adding a bee to propolis.   The method for enhancing the antioxidant activity of propolis according to claim 4, wherein 6 to 240 parts by weight of bees are added to 1 part by weight of propolis.   The method for enhancing the antioxidant activity of propolis according to claim 4 or 5, further comprising adding royal jelly.