JP5765879B2 - Composition for oral consumption containing bee, propolis, and royal jelly - Google Patents

Description

  The present invention relates to a composition for ingestion comprising bees, propolis, and royal jelly.

  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 maintaining health and beauty, and a search for materials having various antioxidative actions has been actively conducted.

  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 of color and flavor, lowering of nutritional value, etc. 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”. Bee pups are foods having a specific amino acid composition and have been reported to have a fatigue recovery effect and the like (Patent Documents 1 to 3).

  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 external preparations for skin such as cosmetics and quasi drugs (Patent Documents 4 to 7).

  It is known that propolis, which is a sticky substance used as a material for beehives, contains various sap activities including antioxidants as a component of plant sap collected by bees from the field (Patent Documents 8 to 8). 11). For this reason, propolis is used as a raw material for health foods.

JP 2002-17296 A JP 2006-197911 A JP 2001-204421 A JP 2001-292712 A JP 2002-20226 A JP 2003-211981 A JP 2006-280249 A JP 2000-44479 A JP 2002-235084 A JP 2002-306092 A Japanese Patent Laid-Open No. 2005-220087

  There is a need for a composition that has an antioxidant activity for protecting the body from oxidative stress, and that can suppress the oxidation of a composition for oral intake and is safe and can be taken on a daily basis as a food. An object of the present invention is to provide a composition that has a synergistic antioxidant activity by including a combination of specific raw materials and can be safely ingested as a food. A further object of the present invention is to provide a composition that exhibits an anti-aging effect, a skin beautifying effect, and an improvement / relief effect of various symptoms caused by menopause based on excellent antioxidant activity.

  As a result of diligent research to solve the above problems, the present inventors have found a good antioxidant effect and anti-aging effect in a composition containing bee, propolis, and royal jelly, and have completed the present invention. .

  According to one aspect of the present invention, a composition for ingestion is provided comprising a bee cub, propolis, and royal jelly.

  According to another aspect of the present invention, there is provided a food or drink comprising bees, propolis, and royal jelly.

  The composition for oral consumption of the present invention has an excellent antioxidant action. Thereby, discoloration of the composition for oral intake, deterioration of flavor, and reduction in nutritional value can be prevented, and the effect of the active ingredient contained in the composition for oral intake can be stabilized for a long period of time. Various injuries caused by active oxygen present in the body (myocardial infarction, arteriosclerosis, diabetes, cancer, stroke, cataract, stiff shoulder, coldness, hypertension, senile) Can treat, improve and / or prevent dementia, spots, freckles, wrinkles, melasma, dullness, and other skin weakness; skin firmness, skin gloss, makeup paste, skin texture, Can improve and / or maintain skin elasticity; can improve and / or prevent skin roughness, dullness, skin pimples / acne; and symptoms of menopause such as face Hot flashes, sweatyness, coldness in the hips and limbs, palpitation, shortness of breath, dizziness, sleep disorders (bad sleep, light sleep, etc.), anxiety, mood instability, reverse feeling, depression, headache, headache, nausea Fatigue, stiff shoulders Back pain, it is possible to improve and / or prevention of such limb pain.

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. It is a graph which shows the menopausal index (SMI) variation | change_quantity of the administration group (test food group) and the placebo group of the composition of this invention. An asterisk (*) in the figure indicates a significant difference p <0.05 with respect to placebo.

  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, 10-24 days after hatching, preferably 20-22 days after hatching Can be used.

  The collected bees can be used, for example, by drying, pulverizing and powdering. For example, bees prepared by the method disclosed in Japanese Patent Application Laid-Open No. 2002-17296 (paragraph [0006]) can be used. A child powder may be used in the present invention. In one aspect of the present invention, the bees frozen at −80 ° C. to −5 ° C. after collection are naturally thawed before use, and after mixing with a stirrer, the resulting mixture is boiled and sterilized for 10 to 120 minutes, The powder obtained by sieving (10-200 mesh) after lyophilization 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. As excipients that can be used in the present invention, excipients that can be used in medicine or food, for example, dextrin, corn starch, rice flour, lactose, potato starch, reduced starch saccharified product, silicon dioxide and the like can be used.

  The propolis used in the present invention is not particularly limited as long as it is a propolis commonly used in foods. For example, honey bees (especially bees and subspecies thereof (Apis melifera, Apis mellifera adanoniii, Apis mellifera scutellata). Propolis taken from)) can be used. Propolis origin plants are not particularly limited, and for example, propolis originating from plants such as baccaris, 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 Nisshin Honey Co., Ltd. can be used in the present invention.

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

  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 powder obtained by obtaining a solid substance, optionally adding an excipient to the solid substance (propolis extract), then spray-drying, granulating and sieving can be used as the propolis of the present invention.

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

  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 Labeling of Royal Jelly” (prepared by the Royal Jelly Fair Trade Council). "Raw royal jelly", "dried royal jelly" and "prepared royal jelly" can be used, both of which include, for example, bees of the honey bee family (especially Apis melifera, Apis mellifera adanoni, Apis) Royal jelly collected from Mellifera scutellata)), preferably from honeybees 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 thawing a stock solution of royal jelly frozen at −80 to −5 ° C. after collection before use and sieving (10 to 200 mesh) after lyophilization is used in 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 may be used. it can.

  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 composition for oral consumption containing 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 per 1 part by weight of propolis, The amount is more preferably 12 parts by weight or more, further 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 is in this range, it can be set as the composition for oral intake which has the outstanding antioxidant effect.

  Each raw material used in the present invention contains the following 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% by weight is contained in a dried bee child powder.

  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 composition for oral consumption of the present invention is, for example, 0.0005% by weight or more, particularly 0.001% by weight or more. 1% by weight or less, particularly 0.05% by weight or less. Further, the amount of p-coumaric acid derived from propolis contained in the composition for oral consumption 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, In particular, it is 0.002% by weight or less. Furthermore, the amount of decenoic acid derived from royal jelly contained in the composition for oral consumption 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%. 0.08% by weight or less.

  In addition, each component in the composition for oral intake of this invention can be measured by a method well-known to those skilled in the art, and a compounding quantity can be computed based on the measured value.

  In a further aspect of the present invention there is provided a composition for oral consumption in the form of a food or drink comprising bees, propolis, and royal jelly.

  The composition for oral consumption in the form of a food or drink according to the present invention can be provided as it is or prepared as a food (health food (nutrient functional food, food for specified health use, supplement, etc.), food for the sick, etc.). .

  The composition for oral consumption of the present invention contains bees, propolis and royal jelly (preferably 0.0003 wt% or more as an ascorbic acid equivalent), and does not contain a known antioxidant such as ascorbic acid or tocopherol. It has a very good antioxidant effect, is safe, and has excellent long-term stability in terms of color, flavor and nutritional value. In addition, since the composition for oral intake of the present invention has high antioxidant activity, various disorders (myocardial infarction, arteriosclerosis) caused by active oxygen present in the living body due to the intake of the composition for oral intake. For improvement and / or prevention of diabetes, cancer, stroke, cataract, stiff shoulders, coldness, hypertension, senile dementia, spots, buckwheat, wrinkles, melasma, dullness, other skin deterioration, etc .; Skin gloss, makeup paste, skin texture, skin elasticity improvement and / or maintenance; skin dullness, skin roughness, skin breakout / acne improvement and / or prevention; and menopause disorders Symptoms such as hot flashes in the face, sweat, low back and limbs, palpitation, shortness of breath, dizziness, sleep disorders (bad sleep, light sleep, etc.), anxiety, mood instability, reverse feeling, depression, Headache, headache, nausea, fatigue , It can be taken stiff shoulders, lower back pain, for the improvement and / or prevention of such limb pain.

  The composition for oral intake of the present invention can be blended with components such as conventionally known colorants, preservatives, fragrances, flavoring agents, and coating agents, if necessary.

  In addition, the composition for oral intake of the present invention can contain a processed plant product, a functional component, or a medicinal component. In addition, these components may be used alone or in combination with the composition for oral intake, or two or more kinds may be used in combination with the composition for oral intake.

  Examples of plants that can be used in combination include plant extracts having an antioxidant action and the like (for example, red-crowned wrinkles, oysters, mulberries, hypericum perforatum, camellia, arabesque garage, yaba santa, rosehip, Plants such as Asunaro, Azalea, Itadori, Ichiakuso, Apricot, Keikanka, Hakukayumatou, Birch, Hawthorn, Achillea millefolium, Tarayou, Dokudami, Tormentilla, Bakumondo, Hiba, Grape, Mukuroji, Mokka, Ganoderma, Roman cutlet, etc. Extracts (eg, grape seeds, hijiki, moroheiya, hamasge, cereals (eg, beans, rice and wheat), herbs (eg, myrrh, basil, thyme, etc.), cacao extracts, Hiekisu, plum extract, hawthorn extract, and plant extracts such as Chlorella); soybean extract having estrogenic or black cohosh extract; blood pressure lowering effect and rice germ extract having tranquilizers and the like.

  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 hypoglycemic agents include, but are not limited to, indigestible dextrin, guava leaf, wheat albumin, L-arabinose, bean extract, mulberry leaf, ginger, salacia, α-linolenic acid, macaque, 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, Gua Gum, Chondroitin Sulfate, Mackerel, Alfalfa, Ginkgo, Psyllium, Barley, Oats, Olives, Gajutsu, 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.), and the like. 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 order to facilitate continuous ingestion, the composition for oral ingestion according to the present invention is, for example, granules (including dry syrup), capsules (soft capsules, hard capsules), tablets (including chewable agents), It is desirable to prepare in the form of various solid preparations such as powders (powder) and pills, or liquid preparations such as liquids for internal use (including liquids, suspensions and syrups). The tablet form is preferable from the viewpoint of stability of each component and ease of ingestion, but is 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 when preparing in other forms, conventional methods may be followed.

  Furthermore, the composition for oral consumption of the present invention in the form of granules, powders, liquids, etc., for example, various foods and drinks such as beverages, confectionery, breads and soups; various pet foods such as dog foods and cat foods, etc. It can also be added to and 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 intake 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, based on an adult weighing 60 kg, a total of 100 to 3000 mg / day, preferably 300 to 2000 mg / day, more preferably 500 to 1000 mg / day of bee pups, propolis and royal jelly is used as a guide. .

  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 naturally thawed before use were mixed with a stirrer and then sterilized by boiling for 30 to 60 minutes. After lyophilization, sieving (50 mesh) was performed to obtain a bee powder. The bee powder was used in the following tests (“antioxidation activity measurement test”, “effectiveness confirmation test in humans”, “mouse skin layer thickness measurement test”, etc.).

  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 beehive box 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 Extract). A dextrin agent having the same weight as that of the solid material was added to the obtained solid material 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 3 and 5 in the “antioxidant activity measurement test” indicates the amount of propolis extract contained in the propolis powder (propolis extract content: 50%). 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 Kanto Branch of the Analytical Chemistry Society of Japan), and the decenoic acid content was measured. Was 5.9% by weight (per dry powder weight).

[Test Example 1] Antioxidant activity measurement test [Preparation of sample solution]
Hereinafter, according to Tables 1 to 5, 100 μL is taken from the dimethyl sulfoxide solution of each sample powder prepared to have a predetermined concentration, and after centrifugation (5000 rpm × 5 minutes), 10 μL is taken from the supernatant, As added.

[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 the concentration (% by weight) of L-ascorbic acid, Abs represents the absorbance of the sample at 550 nm, and (Bla-Abs (550 nm)) represents (blank absorbance measurement value-sample absorbance measurement 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 measurement test of bee, propolis, and royal jelly
Antioxidant activity was measured using each of the bee powder, propolis powder, and royal jelly powder as sample solutions, and the L-ascorbic acid equivalent was calculated. The results are shown in Tables 2-4.

  From the results in Table 2, it was confirmed that the bees had antioxidant activity. Propolis was also confirmed to have an antioxidant effect (Table 3). On the other hand, the antioxidant effect of royal jelly was very low (Table 4).

Antioxidant activity measurement test of a mixture of bee, propolis and royal jelly Antioxidant activity was measured using a mixture of bee powder, propolis powder and royal jelly powder as a sample. The results are shown in Table 5. The theoretical values in the table are the sum of Asc (%) [EQ] measured values (Tables 2 to 4) when bee, propolis, and royal jelly are used alone at each concentration. Asc (%) [EQ] of the measured value and the ratio of the theoretical value.

  In Table 5, in the case of containing 6 to 240 parts by weight of bee powder per 1 part by weight of propolis extract, or in the case of containing 0.3 to 60 parts by weight of bee powder per 1 part by weight of royal jelly powder The antioxidant effect exceeding the theoretical value was confirmed. From these results, it was found that the antioxidant activity is synergistically increased by mixing these three components.

  Here, ascorbic acid, which is generally used as an antioxidant for foods and beverages and cosmetics and its raw materials, for example, is oxidized at about 0.0003 to 0.0005% by weight when injected or blended into food materials. It is effective as an inhibitor (see Food Additives Official Document (8th Edition), Akio Tanimura, Yodogawa Shoten, 2007).

  As a result of this test, the composition for oral consumption containing bee, propolis, and royal jelly has excellent antioxidant activity similar to L-ascorbic acid, and the antioxidant capacity of the composition for oral consumption itself. In addition, the composition was found to be useful as a composition for oral consumption for treating, improving and / or preventing various disorders such as skin caused by oxidation.

[Test Example 2] Efficacy confirmation test in humans By taking a composition having an antioxidant activity, various disorders caused by oxidation can be treated, improved and / or prevented. 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 raw material 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 skin irritation, skin luster, makeup paste, skin roughness, skin texture, skin dullness, skin pimples and acne all after one month of ingestion compared to the time of ingestion. There was a tendency for evaluation to increase in the items.

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

[Measuring method]
The measurement was carried out after washing the face and acclimatized for 20 minutes in a room adjusted to 60% humidity and 25 ° C., and the elasticity value was measured using a Cutometer MPA580 (Courage + Kazaka electronic) according to the attached manual. . The higher the skin elasticity, the higher the Uf value. The moisture value was measured as electrical conductivity (μS) using Skicon 200EX (manufactured by IBI S Co., Ltd.) according to the attached manual. The higher the moisture content on the skin surface, the higher the electrical conductivity.

  The results are shown in FIG. 1 and FIG. The skin elasticity was significantly improved at the first and second months of ingestion compared to the start of ingestion (p <0.05). Although there was no significant difference in the water level, an increasing tendency was confirmed one month after ingestion.

  The above test revealed that the composition for oral consumption 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 composition of the present invention on the skin tissue in more detail, the following feeding test was conducted using mice.

[Preparation of sample]
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 bee pups, royal jelly and propolis mixture was significantly increased compared to group B and group C mice fed the diet containing the same amount of protein. did. Moreover, it can be seen from microscopic observation that the thickness of the dermis layer containing a large amount of collagen is increased in the group A mice.

[Test Example 4] Effect confirmation test for menopausal symptoms In order to confirm the effect of the intake of the composition of the present invention on menopausal symptoms, 25 female monitors aged 40 to 50 years old were tested food group 12 people, placebo group 13 people Divided into. Tablets prepared by the above preparation method with the formulation of Table 6 above were used as test foods. Placebo tablets were prepared in the same manner as the test food with the formulation shown in Table 10 below.

  Subjects continued taking 8 tablets of any tablet daily for 8 weeks. The simple menopause index (SMI) questionnaire shown in Table 11 was conducted before intake, 4 weeks after the start of intake, and 8 weeks after the start of intake (Takao Koyama, Takeshi Aso, Ayumi of Obstetrics and Gynecology Kampo Studies, 1992; 9 Volume: 30-34; Tsutomu Doji, Hideki Mizunuma, Toshinori Fujino, Kenzo Okura, Journal of Obstetrics and Gynecology, September 2007, Vol. 59, No. 9, N-482-N-487). The total points were calculated from the questionnaire results in Table 11.

  For those who showed a menopause tendency in the pre-ingestion assessment (determined according to the climacteric index judgment table in Table 12 except “normal”, score 26 points or more; 8 test food groups, 9 placebo groups) The data after 4 weeks of intake and after 8 weeks of intake were compared. The amount of change in numerical values based on the score before ingestion is shown in Table 13 and FIG.

  From Table 13 and FIG. 5, it was confirmed that the test food group significantly improved menopausal symptoms as compared to the placebo group 4 weeks and 8 weeks after ingestion.

[Prescription example]
Formulation Examples 1 to 13 (tablets)
Compositions having the formulations shown in Table 14 (Prescription Examples 1 to 13) 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 then tableted according to a conventional method to prepare a tablet.

Formulation Examples 14 to 26 (granule)
Compositions (formulation examples 14 to 26) comprising the formulations described in Table 15 were prepared as granules according to conventional methods. 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 27-34 (Liquid)
Compositions (formulation examples 27 to 34) having the formulation described in Table 16 were prepared as liquids according to a conventional method. Specifically, each component was blended according to the formulation and mixed according to a conventional method to form a liquid preparation.

Claims (2)

A composition for oral consumption comprising bee pups, propolis, and royal jelly, comprising 16 to 240 parts by weight of bee pups per part by weight of propolis.   The composition for oral ingestion according to claim 1, comprising 0.3 to 60 parts by weight of bees per 1 part by weight of royal jelly.

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