WO2019230907A1 - 肥満抑制剤、抗認知症剤、消臭剤、抗老化剤、抗糖化剤、抗i型アレルギー剤、抗高血圧剤、風味向上剤、筋肉増強剤、及び骨代謝改善剤 - Google Patents
肥満抑制剤、抗認知症剤、消臭剤、抗老化剤、抗糖化剤、抗i型アレルギー剤、抗高血圧剤、風味向上剤、筋肉増強剤、及び骨代謝改善剤 Download PDFInfo
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- bagasse
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
Definitions
- the present invention relates to an obesity inhibitor, an antidementia agent, a deodorant, an anti-aging agent, an anti-glycation agent, an anti-type I allergic agent, an antihypertensive agent, a flavor enhancer, a muscle enhancer, and a bone metabolism improver.
- Obesity especially obesity due to accumulation of visceral fat, has become a global health problem as a cause of metabolic syndrome.
- the mechanism of obesity is examined systematically, it indicates a state in which fat cells accumulate a large amount of fat and enlarge, and further, the number of enlarged fat cells greatly increases.
- Patent Document 1 discloses that a kiwi extract is effective in suppressing obesity.
- dementia refers to a state in which various obstacles occur due to death of brain cells due to various causes, or work becomes difficult, resulting in difficulties in living. Physical function may also be lost due to atrophy of the entire brain with the onset of dementia.
- Patent Document 2 discloses that royal jelly exhibits antidementia activity.
- Patent Document 3 discloses a deodorant containing a sweet potato-derived distillate as an active ingredient.
- the skin has a three-layer structure of epidermis, dermis, and subcutaneous tissue.
- type I collagen gathers to form a bundle, thereby playing a role as a strut of the dermis, and there are components such as elastin and hyaluronic acid around it.
- these components are reduced or decomposed due to aging or irradiation with ultraviolet rays, the skin tension and gloss are lost, and wrinkles are easily formed.
- Patent Document 4 discloses an anti-aging agent characterized by containing an extract from straw and / or oil tea as an active ingredient.
- Patent Document 5 discloses an anti-aging agent containing an extract of one or more kinds of plants selected from a plant belonging to the genus Macchiaceae.
- Patent Document 6 discloses an anti-aging agent characterized by containing an extract of Ibusuki Sou as an active ingredient.
- Patent Document 7 discloses a collagen production promoter, collagenase inhibitor, and elastase inhibitor characterized by containing an extract from a carambola leaf as an active ingredient.
- saccharification is also called Maillard reaction, and is a non-enzymatic chemical reaction between amino acid or protein and reducing sugar discovered by French scientist L.C.LMaillard in 1912. Saccharification has attracted attention in the field of food chemistry, such as changes in color, aroma, or flavor that occur during the heating of food.
- Patent Document 8 discloses an anti-glycation agent containing a concentrated extract of shochu residue moromi as an active ingredient.
- allergy is defined as “systemic or local damage to a living body based on an immune response”. Allergic reactions are classified into types I to IV. Among them, type I, type II and type III allergies are humoral immunity involving serum antibodies, and type IV allergy is cellular immunity due to sensitized lymphocytes.
- Type I allergy is also called immediate allergy or anaphylactic allergy. Symptoms of type I allergy include hay fever and urticaria. However, since the population having these symptoms is relatively large, an effective anti-type I allergic agent is required.
- Patent Document 9 discloses a cosmetic or skin external preparation having an antiallergic effect by using a carnitine derivative and / or the carnitine derivative in combination with an effect promoter.
- High blood pressure is a state in which blood pressure exceeding the normal range is maintained.
- High blood pressure is one of lifestyle-related diseases, and causes major complications in major organs such as the brain, heart, and kidney, and is therefore a major problem.
- Patent Document 10 discloses an antihypertensive agent containing a mung bean protein degradation product.
- Patent Document 11 is a flavor improving agent for foods and drinks containing a sweet potato-derived distillate as an active ingredient, and a distillate obtained by distilling sweet potato soup is filled with a synthetic adsorbent as a fixed carrier.
- a flavor improver characterized by being a fraction obtained by passing through a column and eluting the component adsorbed on the synthetic adsorbent with a solvent selected from water, ethanol and a mixture thereof is described. ing.
- Patent Document 12 discloses a method of adding a sweet potato-derived extract as an active ingredient to a food or drink, and improving the flavor of the food or drink, wherein the sweet potato-derived extract contains a sweet potato juice, a sweet potato solvent extract, and It is a fraction obtained by processing raw materials selected from sugar cane derived from sugar cane by column chromatography using a fixed carrier, and the improvement of the flavor of food and drink is improved, the salt flavor of salt-containing food and drink is improved, and the egg content
- the method described above is one of improving the egg flavor of a food or drink and improving the flavor of a flavor-containing food or drink.
- muscle is formed by differentiation of muscle cells.
- myoblasts which are progenitor cells, proliferate to a certain number of cells, move to a predetermined muscle region, and fuse with each other to form differentiated myotube cells.
- genes representing muscle-specific traits such as muscle contraction proteins and specific enzymes are expressed in myotubes, and a muscle contraction apparatus is constructed.
- the mitochondria in muscle cells are closely related to muscle functions.
- the main function of mitochondria is energy (ATP) production in the TCA cycle.
- ATP energy
- Patent Document 13 discloses a muscle differentiation induction promoter containing a rice bran and / or rice germ fermentation extract as an active ingredient.
- Patent Document 14 discloses a method of activating mitochondria during exercise using glutathione.
- bone is reconstructed (bone remodeling) by bone metabolism that repeats “bone formation” by osteoblasts (cells that make bones) and “bone resorption” by osteoclasts (cells that break bones), Bone mass is maintained by the balance of these two cells.
- Osteoblasts are derived from mesenchymal stem cells as well as fat and muscle cells, while osteoclasts are derived from blood cells as well as erythrocytes and leukocytes.
- Bone metabolism is initiated by the appearance of a membrane protein called RANKL on the surface of osteoblasts.
- RANKL binds to a receptor called RANK, which is a blood cell
- the blood cell differentiates into an osteoclast.
- Differentiated and mature osteoclasts break bone (bone resorption) and then form the same amount of bone as osteoblasts are absorbed.
- bone mass bone density
- Patent Document 15 discloses an osteogenesis promoter containing as an active ingredient at least one of an acai extract or a matababi extract.
- JP 2001-299264 A Japanese Patent Laid-Open No. 2003-265135 JP 2017-193947 A JP 2018-27983 A JP-A-2018-150240
- the present invention is a novel anti-obesity agent, antidementia agent, deodorant, anti-aging agent, anti-glycation agent, anti-type I allergic agent, anti-hypertensive agent, flavor enhancer, muscle enhancer, and bone metabolism improver
- the purpose is to provide an agent.
- the inventors of the present invention conducted an in vitro test using a bagasse degradation extract that suppresses obesity, anti-aging, anti-glycation, anti-type I allergic, anti-hypertensive, muscle-enhancing, and bone metabolism improvement. It was found to have an action.
- a bagasse degradation extract has an effect of improving short-term memory impairment caused by accumulation of amyloid ⁇ protein by an in vivo test.
- the bagasse decomposition extract has an excellent deodorizing action, in particular, a deodorizing action on food and drink. Furthermore, the present inventors have also found that the bagasse decomposition extract has an effect of improving the flavor of food and drink different from the components described in Patent Documents 11 and 12.
- 1st aspect of this invention provides the obesity inhibitor which contains the decomposition extract of bagasse as an active ingredient as one aspect
- the obesity inhibitor of the present invention may be based on at least an action of inhibiting fat accumulation.
- the obesity inhibitor of the present invention has at least an action of suppressing the accumulation of fat in fat cells. Since suppression of excessive fat accumulation can be an effective means for preventing obesity, obesity is suppressed by the obesity inhibitor of the present invention.
- the first aspect of the present invention provides a fat accumulation inhibitor containing a bagasse decomposition extract as an active ingredient.
- the second aspect of the present invention provides an antidementia agent containing a bagasse decomposition extract as an active ingredient.
- the second aspect of the present invention can also be said to provide a short-term memory disorder improving / suppressing agent containing a bagasse decomposition extract as an active ingredient.
- the third aspect of the present invention provides a deodorant containing a bagasse decomposition extract as an active ingredient.
- the fourth aspect of the present invention provides an anti-aging agent containing a bagasse decomposition extract as an active ingredient.
- the anti-aging agent of this invention is excellent in the anti-aging effect by including the decomposition extract of bagasse as an active ingredient.
- the fourth aspect of the present invention can also provide an extracellular matrix degrading enzyme inhibitor containing a bagasse degradation extract as an active ingredient as another embodiment.
- the fourth aspect of the present invention can also be said to provide a fibroblast activator as still another embodiment.
- the fifth aspect of the present invention relates to an anti-glycation agent containing a bagasse decomposition extract as an active ingredient. Since the anti-glycation agent of the present invention contains a bagasse degradation extract as an active ingredient, it has excellent anti-glycation activity.
- the anti-glycation agent is excellent in anti-glycation activity, it can be suitably used as a food or drink for anti-glycation.
- the sixth aspect of the present invention provides, as one aspect, an anti-type I allergic agent containing a bagasse decomposition extract as an active ingredient.
- an anti-type I allergy agent of the present invention the symptoms of type I allergy can be suppressed (treated, alleviated or prevented).
- the anti-type I allergic agent of the present invention may be based on the degranulation inhibitory action of mast cells or basophils.
- the mechanism of type I allergic reaction is as follows. (1) When an antigen (allergen) such as pollen or mite enters the living body, a helper T cell (Th2 cell) issues a command to differentiate the B cell into an immunoglobulin E (IgE) antibody-producing cell. (2) IgE antibody specific to the antigen is produced from IgE antibody-producing cells. (3) An IgE antibody binds to mast cells or basophils, and an antigen binds there again, whereby chemical mediators such as histamine and leukotriene are secreted (degranulated) and allergic symptoms appear.
- allergen allergen
- Th2 cell helper T cell
- IgE antibody specific to the antigen is produced from IgE antibody-producing cells.
- An IgE antibody binds to mast cells or basophils, and an antigen binds there again, whereby chemical mediators such as histamine and leukotriene are secreted (degranulated) and allergic symptoms appear.
- the anti-type I allergic agent of the present invention has at least an action of suppressing degranulation of mast cells or basophils. Therefore, according to the anti-type I allergy agent of the present invention, the symptoms of type I allergy can be effectively suppressed.
- the sixth aspect of the present invention can also be said to provide a degranulation inhibitor for mast cells or basophils, which contains a bagasse degradation extract as an active ingredient, as another embodiment.
- the seventh aspect of the present invention provides, as one aspect, an antihypertensive agent containing a bagasse decomposition extract as an active ingredient.
- the antihypertensive agent of this invention is excellent in the antihypertensive effect by including the decomposition extract of bagasse as an active ingredient.
- the seventh aspect of the present invention can also provide an angiotensin converting enzyme inhibitor containing a bagasse degradation extract as an active ingredient as another embodiment.
- the eighth aspect of the present invention provides a flavor improver containing a bagasse decomposition extract.
- the flavor improver may be a flavor improver that enhances the preferred flavor of food and drink.
- the flavor enhancer may be a flavor enhancer that reduces the taste of food and drink.
- the eighth aspect of the present invention can also be said to provide a food or drink containing the flavor improver.
- the eighth aspect of the present invention can also be said to provide a preferred flavor enhancer for foods and drinks containing a bagasse decomposition extract.
- the eighth aspect of the present invention can also be said to provide a taste-reducing agent for foods and drinks containing a bagasse decomposition extract.
- the ninth aspect of the present invention provides a muscle strengthening agent containing a bagasse decomposition extract as an active ingredient.
- the ninth aspect of the present invention provides a myotube differentiation promoting agent containing a bagasse degradation extract as an active ingredient. It can also be said that the ninth aspect of the present invention provides a mitochondrial activator containing a bagasse degradation extract as an active ingredient.
- the tenth aspect of the present invention provides a bone metabolism improving agent containing a bagasse decomposition extract as an active ingredient.
- the tenth aspect of the present invention provides an osteogenesis promoter containing a bagasse decomposition extract as an active ingredient.
- the 10th side surface of this invention can also be said to provide the bone resorption inhibitor containing the decomposition extract of bagasse as an active ingredient.
- the bagasse decomposition extract in the present invention may be a decomposition treatment liquid obtained by at least one decomposition treatment selected from the group consisting of alkali treatment, hydrothermal treatment, acid treatment, subcritical water treatment and explosion treatment.
- the decomposition extract of bagasse may be a fraction obtained by passing the decomposition treatment liquid through a column packed with a fixed carrier.
- the fixed carrier is preferably a synthetic adsorbent or an ion exchange resin.
- the bagasse decomposition extract elutes the components adsorbed on the synthetic adsorbent with at least one solvent selected from the group consisting of water, methanol, ethanol and mixtures thereof. It is also possible to obtain a fraction obtained by making it occur.
- the synthetic adsorbent is preferably an aromatic resin, an acrylic acid-based methacrylic resin, or an acrylonitrile aliphatic resin.
- the decomposition extract of bagasse is obtained by passing the decomposition treatment liquid through a column packed with a synthetic adsorbent as a fixed carrier and eluting the components adsorbed on the synthetic adsorbent with a mixed solvent of ethanol and water.
- the synthetic adsorbent is an unsubstituted aromatic resin
- the column temperature is 20 to 60 ° C.
- the volume ratio of ethanol and water may be 50/50 to 60/40.
- a novel anti-obesity agent, antidementia agent, deodorant, anti-aging agent, anti-glycation agent, anti-type I allergy agent, anti-hypertensive agent, flavor enhancer, muscle enhancer, and bone Metabolism improving agents can be provided.
- FIG. 5 is a photomicrograph showing the results of staining accumulated lipid droplets in the adipocytes of progenitor cells (undifferentiated a1-1), positive control a1-1, comparative example a1-1 and example a1. It is a graph which shows the fat accumulation rate of comparative example a1-1, positive control a1-1, and Example a1.
- 6 is a micrograph showing the result of staining accumulated lipid droplets in the adipocytes of progenitor cells (undifferentiated a1-2), Comparative Example a1-2, and Example a2. It is a graph which shows the fat accumulation rate of comparative example a1-2 and Example a2.
- Bagasse typically refers to bagasse discharged in the sugar production process in the raw sugar production process.
- the bagasse discharged in the sugar production process in the raw sugar factory includes not only the final bagasse that has left the final press, but also shredded sweet potato that has been bitten by the subsequent press including the first press.
- a suitable bagasse is bagasse discharged after squeezing the sugar juice in the raw sugar factory.
- the bagasse has different moisture content, sugar content and composition ratio depending on the type of sweet potato, harvest time, etc., but in the present invention, these bagasse can be used arbitrarily.
- the raw material bagasse as in the raw sugar factory, for example, the bagasse remaining after the sweet potato pressing discharged in the brown sugar manufacturing factory, or the sugar solution was pressed from the sweet potato by small-scale implementation at the laboratory level Later bagasse can also be used.
- the bagasse decomposition extract may be a decomposition solution of bagasse (and / or a processed product thereof).
- the decomposition treatment liquid can be obtained by at least one decomposition treatment selected from the group consisting of alkali treatment, hydrothermal treatment, acid treatment, subcritical water treatment, and explosion treatment.
- the bagasse decomposition treatment herein requires that some or all of the chemical structure of lignin, cellulose, and / or hemicellulose be broken.
- the decomposition treatment is preferably alkali treatment or hydrothermal treatment from the viewpoint of easily obtaining a bagasse decomposition extract.
- the alkali treatment may be a treatment in which an alkaline solution is brought into contact with bagasse.
- Examples of the method of bringing the alkaline solution into contact include a method of sprinkling an alkaline solution over bagasse, a method of immersing bagasse in an alkaline solution, and the like.
- the mixture of bagasse and alkaline solution may be immersed while stirring.
- alkaline solution examples include a sodium hydroxide aqueous solution, a potassium hydroxide aqueous solution, and an ammonia aqueous solution. These alkaline solutions may be used alone or as a mixture of two or more of these solutions.
- the alkaline solution is preferably an aqueous sodium hydroxide solution from the viewpoint of being inexpensive and easily used in the food production process.
- the temperature (liquid temperature) of the alkaline solution is preferably 40 ° C. or higher, more preferably 100 ° C. or higher, and further preferably 130 ° C. or higher, from the viewpoint of shortening the processing time of the decomposition treatment.
- the temperature of the alkaline solution is preferably 250 ° C. or less, more preferably 200 ° C. or less, and further preferably 150 ° C. or less, from the viewpoint of preventing polysaccharides from remaining in the decomposition treatment solution.
- the temperature of the alkaline solution is 40 to 250 ° C, 40 to 200 ° C, 40 to 150 ° C, 100 to 250 ° C, 100 to 200 ° C, 100 to 150 ° C, 130 to 250 ° C, or 130 to 200 ° C, 130 to 150 ° C. It may be ° C.
- the alkali treatment may be performed under normal pressure or may be performed under pressure.
- the pressure may be 0.1 MPa or more, or 0.2 MPa or more, and may be 4.0 MPa or less, 1.6 MPa or less, or 0.5 MPa or less.
- the pressure is 0.1-4.0 MPa, 0.1-1.6 MPa, 0.1-0.5 MPa, 0.2-4.0 MPa, 0.2-1.6 MPa, or 0.2-0. It may be 5 MPa.
- Hydrothermal treatment may be a treatment in which bagasse is brought into contact with hot water or water vapor under high pressure. More specifically, the hydrothermal treatment may be, for example, a method in which water is added so that the solids concentration of bagasse becomes 0.1 to 50% and the decomposition treatment is performed under high temperature and high pressure conditions.
- the temperature of water or water vapor is preferably 130 to 250 ° C., and the pressure applied is preferably 0.1 to 0.5 MPa higher than the saturated water vapor pressure of water at each temperature.
- the acid treatment may be a treatment in which an acidic solution is brought into contact with bagasse.
- the acidic solution include dilute sulfuric acid.
- the method of bringing the acid solution into contact with the bagasse, the temperature of the acid solution in the acid treatment, and the pressure condition in the acid treatment may be the same as the method or conditions in the alkali treatment described above.
- the subcritical water treatment may be a treatment in which subcritical water is brought into contact with bagasse.
- the method of bringing the subcritical water into contact with the bagasse may be the same as the method in the alkali treatment described above.
- the conditions for the subcritical water treatment are not particularly limited, but it is preferable that the temperature of the subcritical water is 160 to 240 ° C. and the treatment time is 1 to 90 minutes.
- Blasting treatment involves breaking down bagasse by instantaneously releasing it to atmospheric pressure by, for example, opening a valve provided in the pressure-resistant reaction vessel at once after decomposing insoluble xylan contained in bagasse to some extent by hydrothermal treatment. It may be a process.
- the decomposition treatment liquid after the above-described decomposition treatment, a treatment for separating a solid content and a liquid content may be performed.
- the liquid obtained after the separation can be used as the decomposition treatment liquid.
- the method for separating the solid content and the liquid content may be separation by strainer, filtration, centrifugation, decantation, or the like.
- the decomposition treatment liquid polymer components such as polysaccharides may be removed by membrane separation.
- the liquid after membrane separation can be used as the decomposition treatment liquid.
- the separation membrane is not particularly limited as long as it is an ultrafiltration membrane (UF membrane).
- the molecular weight cutoff of the ultrafiltration membrane is preferably 2500 to 50000, more preferably 2500 to 5000.
- Ultrafiltration membrane materials include polyimide, polyethersulfone (PES), polysulfone (PS), polyacrylonitrile (PAN), polyvinylidene fluoride (PVDF), regenerated cellulose, cellulose, cellulose ester, sulfonated polysulfone, and sulfonation.
- Polyethersulfone, polyolefin, polyvinyl alcohol, polymethyl methacrylate, polytetrafluoroethylene and the like can be used.
- the filtration method of the ultrafiltration membrane may be dead-end filtration or crossflow filtration, but is preferably crossflow filtration from the viewpoint of suppressing membrane fouling.
- the membrane form of the ultrafiltration membrane an appropriate form such as a flat membrane type, a spiral type, a tubular type, and a hollow fiber type can be used. More specifically, SUEZ GE series, GH series, GK series, PW type, HWSUF type, KOCH HFM-180, HFM-183, HFM-251, HFM-300, HFK-131, HFK-328 , MPT-U20, MPS-U20P, MPS-U20S, Sender 1, SPE3, SPE5, SPE10, SPE30, SPV5, SPV50, SOW30, molecular weight 3 of Microza (registered trademark) UF series manufactured by Asahi Kasei Corporation NTR7410, NTR7450, etc. manufactured by Nitto Denko Corporation.
- Microza registered trademark
- the bagasse decomposition extract may be a fraction obtained by passing the decomposition treatment liquid described above through a column packed with a fixed carrier. By passing the decomposition treatment liquid through the column, the active ingredient in the decomposition treatment liquid is adsorbed on the fixed carrier, and most of the sugars and inorganic salts flow out as they are.
- the decomposition treatment liquid can be directly passed through the column, or can be adjusted to an arbitrary concentration with water and passed through the column.
- the pH may be adjusted before passing through the column.
- the decomposition treatment liquid is preferably adjusted to pH 6 or less.
- the pH of the decomposition treatment liquid may be more than 4.5 and 6 or less.
- the decomposition treatment liquid is preferably adjusted to pH 5 or higher.
- the fixed carrier is preferably either a synthetic adsorbent or an ion exchange resin.
- the synthetic adsorbent is preferably a synthetic porous adsorbent.
- An organic resin is preferably used as the synthetic adsorbent (synthetic porous adsorbent).
- the organic resin is preferably at least one selected from the group consisting of aromatic resins, acrylic acid-based methacrylic resins, and acrylonitrile aliphatic resins.
- aromatic resin examples include styrene-divinylbenzene resin.
- aromatic resins include porous resins such as aromatic resins having a hydrophobic substituent, unsubstituted aromatic resins, and aromatic resins specially processed into unsubstituted groups. Of these, an unsubstituted aromatic resin or an aromatic resin specially treated to an unsubstituted group is preferred.
- the amount of the synthetic adsorbent packed in the column can be appropriately determined depending on the size of the column, the type of the synthetic adsorbent, and the like.
- the flow rate when the decomposition treatment liquid is passed can be appropriately changed depending on the size of the column, the type of elution solvent, the type of synthetic adsorbent, etc.
- SV 1-30 hours ⁇ 1 .
- SV Space Velocity, space velocity
- the adsorbed component (active component) adsorbed on the synthetic adsorbent can be eluted with a solvent (elution solvent). From the viewpoint of recovering the adsorbed component more efficiently, it is preferable to wash away saccharides and inorganic salts remaining in the column with water before eluting the adsorbed component.
- the eluted component can be used as a decomposition extract of bagasse.
- the elution solvent may be at least one selected from the group consisting of water, methanol, ethanol, and mixtures thereof.
- the elution solvent is preferably a mixed solvent of alcohol and water, more preferably a mixed solvent of ethanol and water, and a volume ratio of 50/50 to 60/40 (ethanol from the viewpoint that the adsorbed component can be eluted more efficiently at room temperature. More preferred is a mixed solvent of ethanol and water.
- the column temperature at the time of elution may be room temperature, but by increasing the column temperature to a temperature higher than room temperature, ethanol can be mixed in a mixed solvent of ethanol and water. The mixing ratio can be reduced, and the adsorbed component can be eluted more efficiently.
- the temperature is preferably 20 to 60 ° C, more preferably 40 to 60 ° C.
- the inside of the column may be under normal pressure or under pressure.
- the ion exchange resin is classified into a gel type resin and a porous resin such as a porous type, a microporous type or a high porous type based on the form of the resin, but there is no particular limitation.
- the ion exchange resin is preferably an anion exchange resin.
- As the anion exchange resin a strong basic anion exchange resin or a weak basic anion exchange resin may be used.
- a strongly basic anion exchange resin is preferably used, but there is no particular limitation when a decomposition treatment liquid by other treatment is used as a raw material.
- Examples of commercially available strong base anion exchange resins include Diaion (trademark) PA306, PA308, PA312, PA316, PA318L, HPA25, SA10A, SA12A, SA11A, SA20A, UBA120 (above, manufactured by Mitsubishi Chemical Corporation), Amberlite (Trademark) IRA400J, IRA402B1, IRA404J, IRA900J, IRA904, IRA458RF, IRA958, IRA410J, IRA411, IRA910CT (above, manufactured by Organo Corporation), Dowex (trademark) Marathon A, Marathon MSA, MONOSSPHERE550A, Marathon A2 (above, Dow Chemical Japan Co., Ltd.).
- the amount of the ion exchange resin packed in the column can be appropriately determined depending on the size of the column, the type of the ion exchange resin, etc., but it should be 2 to 10,000 times wet volume with respect to the solid content of the decomposition treatment liquid. Preferably, it is 5 to 500 times wet volume.
- the liquid passing conditions can be appropriately set depending on the kind of pretreatment liquid, the kind of ion exchange resin, and the like.
- the liquid flow rate is 100 to 300% by volume of the ion exchange resin
- the column temperature is 40 to 90 ° C.
- the column may be at normal pressure or under pressure.
- the bagasse decomposition extract is obtained by passing through a column packed with ion exchange resin and eluting with an eluent such as an aqueous solution of salt, acid, alcohol or a mixture thereof. It may be a fraction obtained. In this case, the eluent may be deaerated.
- an eluent such as an aqueous solution of salt, acid, alcohol or a mixture thereof. It may be a fraction obtained. In this case, the eluent may be deaerated.
- the bagasse decomposition extract may be a concentrate obtained by concentrating the above-described decomposition treatment liquid or fraction.
- the concentration method may be a known method, for example, a method such as solvent distillation under reduced pressure, freeze drying, or the like.
- the decomposition treatment solution or fraction can be concentrated 15 to 30 times, and the concentrated component can be used as a bagasse decomposition extract.
- the decomposition extract of bagasse can be obtained, for example, as follows. A 1% by mass sodium hydroxide aqueous solution is added to bagasse so that the solid concentration is 0.1 to 50%, and the mixture is boiled at 100 ° C. to obtain a decomposition treatment solution (alkali treatment solution). The decomposition treatment liquid is subjected to ultrafiltration with a UF membrane having a molecular weight cut off of 2500 to 5000, the resulting filtrate is adjusted to acidity, and then the column is packed into a column filled with an unsubstituted aromatic resin. The liquid is passed at a temperature of 20 to 60 ° C.
- the components adsorbed on the column are eluted with a mixed solvent (elution solvent) of ethanol and water having a volume ratio of 50/50 to 60/40 (ethanol / water) at a column temperature of 20 to 60 ° C.
- a mixed solvent elution solvent
- the fraction which the amount of the eluate collected from the elution start time with the mixed solvent of water elutes within 45 times wet volume of the aromatic resin is collected.
- Collect the collected fractions fractions containing components that have an effect of improving bone metabolism
- concentrate by conventional means (evaporation of solvent under reduced pressure, freeze-drying, etc.) to obtain a decomposed extract of bagasse Can do.
- the bagasse decomposition extract thus obtained can be stored as a liquid or powdery extract concentrated so that the solid content is 30% by mass or more. When the extract is liquid, it is preferable to store the extract by refrigeration.
- the bagasse decomposition extract can be obtained as follows. That is, water is added to bagasse so that the solids concentration is 0.1 to 50%, hydrothermal treatment is performed with water at 130 to 250 ° C. under a pressure of 0.2 to 4.0 MPa, and the solid liquid is obtained by filtration. A decomposition treatment liquid (hydrothermal treatment liquid) is obtained by separation. The obtained hydrothermal treatment liquid was passed through a column packed with an aromatic resin specially treated with an unsubstituted group at a temperature of 20 to 60 ° C., and then the components adsorbed on the column were treated with the column temperature.
- Elution is performed with a mixed solvent (elution solvent) of ethanol and water having a volume ratio of 50/50 to 60/40 (ethanol / water) at 20 to 60 ° C. and collected from the start of elution with the mixed solvent of ethanol and water.
- the fraction that elutes within 5% wet volume of the aromatic resin is recovered.
- the bagasse decomposition extract thus obtained can be stored as a liquid or powdery extract concentrated so that the solid content is 30% by mass or more. When the extract is liquid, it is preferable to store the extract by refrigeration.
- the bagasse decomposition extract in each embodiment described above may be liquid or powder.
- the powdered bagasse decomposition extract is produced by using, for example, a liquid bagasse decomposition extract by a spray drying method, a freeze drying method, a fluidized bed granulation method, a powdering method using an excipient, or the like. Can do.
- the decomposition extract of bagasse preferably contains at least one selected from the group consisting of phenylpropanoids such as p-coumaric acid, ferulic acid, caffeic acid and vanillin, and lignin and its decomposition products.
- phenylpropanoids such as p-coumaric acid, ferulic acid, caffeic acid and vanillin, and lignin and its decomposition products.
- the obesity inhibitor in this specification is a composition provided with the effect
- the action of inhibiting obesity is, for example, the action of suppressing fat accumulation in fat cells (fat accumulation inhibiting action), the action of promoting the degradation of fat accumulated in fat cells (fatty degradation promoting action), or the action of fat cells It may be an action of suppressing the growth (adipocyte growth inhibitory action) or the like. That is, the obesity inhibitor in this specification may be a fat accumulation inhibitor, a lipolysis promoter, an adipocyte growth inhibitor, or the like.
- the obesity inhibitor according to one embodiment contains the bagasse decomposition extract described above as an active ingredient.
- the obesity inhibitor according to the present embodiment may be composed only of a decomposition extract of bagasse, which is an active ingredient, and may further contain a material that can be used for foods, quasi drugs, or pharmaceuticals.
- Materials that can be used for foods, quasi drugs or pharmaceuticals are not particularly limited. For example, amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients, binders , Lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- proteins include milk casein, whey, soy protein, wheat protein, egg white and the like.
- carbohydrate include corn starch, cellulose, pregelatinized starch, wheat starch, rice starch, and potato starch.
- examples of the fat include salad oil, corn oil, soybean oil, safflower oil, olive oil, palm oil and the like.
- Sweeteners include, for example, sugars such as glucose, sucrose, fructose, glucose fructose liquid sugar, fructose glucose liquid sugar, sugar alcohols such as xylitol, erythritol, maltitol, artificial sweeteners such as sucralose, aspartame, saccharin, and acesulfame K. And stevia sweeteners.
- Examples of the mineral include calcium, potassium, phosphorus, sodium, manganese, iron, zinc, magnesium, and salts thereof.
- vitamins include vitamin E, vitamin C, vitamin A, vitamin D, vitamin B, biotin, niacin and the like.
- examples of the excipient include dextrin, starch, lactose, crystalline cellulose and the like.
- Examples of the binder include polyvinyl alcohol, gelatin, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and the like.
- Examples of the lubricant include magnesium stearate, calcium stearate, talc and the like.
- Examples of the disintegrant include crystalline cellulose, agar, gelatin, calcium carbonate, sodium bicarbonate, dextrin and the like.
- Examples of the emulsifier or surfactant include sucrose fatty acid ester, citric acid, lactic acid, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, and lecithin.
- Examples of the base include cetostearyl alcohol, lanolin, polyethylene glycol and the like.
- Examples of the solubilizer include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate and the like.
- suspending agent examples include glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxymethyl cellulose, sodium alginate and the like. You may use these individually by 1 type or in combination of 2 or more types.
- the content of the decomposition extract of bagasse as an active ingredient may be appropriately set according to the form and purpose of use of the obesity inhibitor described later.
- it is preferably 100 ⁇ g / g or more, more preferably 250 ⁇ g / g or more, still more preferably 400 ⁇ g / g or more, preferably based on the total amount of the obesity inhibitor. It is 10 mg / g or less, More preferably, it is 7.5 mg / g or less, More preferably, it is 5 mg / g or less.
- the anti-obesity agent may be in any form of solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, capsule, troche Any dosage form such as liquid, suspension, etc. may be used.
- the obesity inhibitor of the present embodiment particularly has an action of suppressing fat accumulation in adipocytes. Therefore, by ingesting the obesity inhibitor of the present embodiment, fat accumulation in fat cells is suppressed, and as a result, obesity is suppressed.
- This obesity suppressant is useful because it can effectively suppress or eliminate obesity without resorting to excessive exercise or dietary restrictions.
- Whether an obesity inhibitor has a fat accumulation-inhibiting action can be determined by, for example, staining fat droplets accumulated during differentiation induction from adipose precursor cells into adipocytes with a lipophilic dye and observing them under a microscope. This can be confirmed by observing whether or not the fat droplets of the specimen to which the fat inhibitor is added are reduced compared to the specimen to which the agent is not added. Whether or not the obesity inhibitor has a fat accumulation inhibitory effect is determined by extracting the lipophilic pigment stained with lipid droplets, measuring the absorbance, and determining the fat accumulation from the change in absorbance associated with the addition of the fat inhibitor. It can also be confirmed by calculating the degree.
- the obesity inhibitor can be used as food, quasi-drug or pharmaceutical.
- the food may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like.
- the obesity inhibitor may be administered parenterally, such as intravenously, or may be administered orally.
- the obesity inhibitor is preferably administered orally.
- the dosage is preferably such that the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, for example, 150 ⁇ g / kg ( (Body weight) is more preferably administered, more preferably 250 ⁇ g / kg (body weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 100 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 500 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and the effect
- the dosage is preferably such that the bagasse degradation extract is 60 ⁇ g / kg (body weight) or more per dose, preferably 120 ⁇ g / kg (body weight) or more. It is more preferable that the dose is 180 ⁇ g / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 180 ⁇ g / kg (body weight) or more per day, more preferably 360 ⁇ g / kg (body weight) or more, and more preferably 540 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and the effect
- the obesity inhibitor of the present embodiment has the above-described action, it can be used for patients diagnosed with obesity and for standard weight persons who desire to prevent obesity.
- the specific aspect of the fat accumulation inhibitor according to one embodiment may be the same as the aspect of the obesity inhibitor described above. That is, the fat accumulation inhibitor according to one embodiment may be obtained by replacing “obesity inhibitor” with “fat accumulation inhibitor” in the above description of the obesity inhibitor.
- One embodiment of the present invention comprises the step of administering to a subject in need thereof an obesity inhibitor containing the above-described decomposition extract of bagasse as an active ingredient, or an effective amount of a fat accumulation inhibitor, Alternatively, it can be regarded as a method of suppressing fat accumulation. Moreover, one Embodiment of this invention can be regarded as the decomposition extract of bagasse for using for the method of suppressing obesity or fat accumulation.
- the subject in the above method may be a mammal, preferably a human.
- the aspect, administration method, dose (intake amount) and the like of the obesity inhibitor or fat accumulation inhibitor may be the same as those described above.
- a further embodiment of the present invention can be regarded as the use of a bagasse decomposition extract for the production of an obesity inhibitor or a fat accumulation inhibitor.
- one embodiment of the present invention can be regarded as the use of a bagasse decomposition extract for suppressing obesity or fat accumulation.
- the aspect of the obesity inhibitor or fat accumulation inhibitor may be the same as described above.
- the anti-dementia agent of the present invention has an anti-dementia action.
- the “anti-dementia effect” in the present invention is a concept including an action to prevent the onset of dementia, an action to delay the onset of dementia, and an action to recover dementia once onset from the onset state. is there.
- the dementia targeted by the antidementia agent of the present invention may be Alzheimer's dementia.
- Alzheimer-type dementia has a pathological condition (choline hypothesis) caused by the loss of acetylcholinergic neurons in the Minelt nucleus of the basal forebrain and a pathological condition caused by accumulation of amyloid ⁇ protein (amyloid hypothesis).
- the dementia targeted by the antidementia agent of the present invention may be Alzheimer type dementia based on any theory, and is preferably Alzheimer type dementia based on the amyloid hypothesis. That is, the dementia targeted by the antidementia agent of the present invention may be Alzheimer type dementia caused by accumulation of amyloid ⁇ protein.
- the present invention relates to an anti-dementia agent for Alzheimer-type dementia, an anti-dementia agent for Alzheimer-type dementia based on the amyloid hypothesis, or an anti-dementia agent for Alzheimer-type dementia caused by accumulation of amyloid ⁇ protein. It can also be provided.
- Alzheimer-type dementia based on the amyloid hypothesis, it is thought that cognitive function is impaired because cranial neurons are killed by accumulation of tau protein in the brain in addition to amyloid ⁇ protein. Accumulation of amyloid ⁇ protein begins in the initial stage, and accumulation of tau protein begins after about 10 years. Thereafter, cerebral nerve cells are killed by the continuous accumulation of amyloid ⁇ protein and tau protein, and dementia is developed about 25 years after the initial stage.
- the antidementia agent of the present invention can also be said to have an action of suppressing accumulation of amyloid ⁇ protein in the brain, an action of reducing amyloid ⁇ protein accumulated in the brain, It can also be said that it has an action of suppressing accumulation of tau protein in the brain and an action of reducing tau protein accumulated in the brain.
- the present invention provides a memory impairment improving / suppressing agent.
- the memory impairment improving / suppressing agent of the present invention has an action of improving / suppressing memory impairment.
- “Improvement / suppression of memory impairment” in the present invention includes an action for preventing the onset of memory disorder, an action for delaying the onset of memory disorder, and an action for recovering a memory disorder once developed from a state at the time of onset. It is a concept.
- the memory disorder targeted by the memory disorder improving / suppressing agent of the present invention may be a long-term memory disorder or a short-term memory disorder, but is preferably a short-term memory disorder. That is, the present invention can be said to provide an agent for improving / suppressing short-term memory impairment, and further can provide an agent for improving / suppressing short-term memory impairment resulting from accumulation of amyloid ⁇ protein.
- the antidementia agent according to one embodiment contains the above-described decomposition extract of bagasse as an active ingredient.
- the antidementia agent may consist only of a decomposition extract of bagasse, which is an active ingredient, and may further contain a material that can be used for foods, quasi drugs or pharmaceuticals.
- Materials that can be used for foods, quasi drugs or pharmaceuticals are not particularly limited. For example, amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients, binders , Lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- the content of the bagasse decomposition extract which is an active ingredient, may be appropriately set according to the form of the antidementia agent described below, the purpose of use, etc. From the viewpoint of more effectively exerting the dementia effect, the following ranges are preferable based on the total amount of the anti-dementia agent.
- the content of the decomposition extract of bagasse is preferably 1% by mass or more, more preferably 3% by mass or more, and further preferably 5% by mass or more as a solid content excluding monosaccharides and oligosaccharides. Moreover, it is preferably 50% by mass or less, more preferably 40% by mass or less, and still more preferably 30% by mass or less.
- An antidementia agent can be used as food, quasi-drug or pharmaceutical.
- the food may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like.
- An antidementia agent can also be used as a feed or feed additive.
- the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured fish and shellfish.
- “Feed” includes anything that animals orally take for nutritional purposes. More specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Is included. Moreover, when classifying from the aspect of a feeding method, it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- the antidementia agent may be in any form such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, capsule, troche Any dosage form such as an agent, solution, suspension, etc. may be used.
- the antidementia agent may be administered parenterally such as intravenous administration, or may be administered orally.
- the antidementia agent is preferably administered orally.
- the dosage is preferably administered so that the decomposition extract of bagasse is 100 ⁇ g / kg (body weight) or more per time, and 150 ⁇ g / kg ( (Weight) or more is more preferable, and administration is more preferably 200 ⁇ g / kg (weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 200 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 400 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and an antidementia effect
- the amount of intake (daily intake or dose) of the product is monosaccharide and low It is preferably 50 to 3000 mg / kg (body weight), more preferably 100 to 2000 mg / kg (body weight), based on the total amount (solid content) of the decomposition extract of bagasse excluding saccharides.
- the above-mentioned product intake (daily intake) is preferably 1 to 1000 mg / kg (based on the total amount of decomposition extract of bagasse excluding monosaccharides and oligosaccharides (solid content)). Weight).
- the antidementia agent of this embodiment can be used for humans or animals in which amyloid ⁇ protein is accumulated.
- the antidementia agent of the present embodiment can be used for humans or animals suffering from dementia (or Alzheimer-type dementia), humans or animals suffering from memory impairment (or short-term memory impairment).
- dementia and memory impairment may be due to accumulation of amyloid ⁇ protein.
- the specific aspect of the short-term memory impairment improving / suppressing agent according to an embodiment may be the same as the aspect of the antidementia agent described above. That is, the short-term memory disorder improving / suppressing agent according to an embodiment may be a term obtained by replacing “an anti-dementia agent” with “short-term memory disorder improving / suppressing agent” in the above description of the antidementia agent. .
- One embodiment of the present invention comprises the step of administering to a subject in need thereof an effective amount of an anti-dementia agent or a short-term memory impairment ameliorating / suppressing agent containing the bagasse degradation extract described above as an active ingredient. It can also be regarded as a method for improving / suppressing dementia or short-term memory impairment. In addition, one embodiment of the present invention can be regarded as a bagasse degradation extract for use in a method for improving / suppressing dementia or short-term memory impairment.
- the subject in the above method may be a mammal, preferably a human.
- the aspect of the antidementia agent or the short-term memory impairment improving / suppressing agent, the administration method, the dose (intake amount) and the like may be the same as those described above.
- a further embodiment of the present invention can also be viewed as the use of a bagasse degradation extract for the manufacture of an antidementia agent or a short-term memory impairment improving / suppressing agent.
- One embodiment of the present invention can also be viewed as the use of bagasse degradation extract to improve / suppress dementia or short-term memory impairment.
- the aspect of the antidementia agent or the short-term memory impairment improving / suppressing agent may be the same as described above.
- the "deodorant” in this specification contains the component (active ingredient) which has a deodorizing effect.
- the deodorant may exhibit a deodorizing effect by erasing or suppressing the odor of the object to be deodorized, and exhibits a deodorizing effect by masking the odor of the object to be deodorized. It may be a thing (masking agent).
- the deodorant which concerns on one Embodiment contains the decomposition extract of bagasse mentioned above as an active ingredient.
- the decomposition extract of bagasse contained in the deodorant may be a decomposition treatment liquid obtained by at least one decomposition treatment selected from the group consisting of alkali treatment, hydrothermal treatment, acid treatment, and subcritical water treatment.
- the deodorant of this embodiment may further contain other components as long as the effects of the present invention are not impaired.
- other components include other deodorants, fragrances, alcohols, surfactants, antibacterial agents, stabilizers, viscosity modifiers, pH adjusters, preservatives, colorants, and the like.
- the deodorant of this embodiment is used as a food (deodorant for food), etiquette deodorant, pet deodorant, environmental deodorant, detergent, softener, hair color agent, permanent agent or cosmetic material.
- a food deodorant for food
- etiquette deodorant pet deodorant
- environmental deodorant detergent, softener
- hair color agent permanent agent or cosmetic material.
- Cosmetic material e.g., etiquette deodorant, pet deodorant, environmental deodorant, detergent, softener, hair color agent, permanent agent or cosmetic material.
- body odor body odor
- hair or Odors attached to the body smell of grilled meat, cigarette odor, etc.
- household garbage or industrial waste storage area household garbage or industrial waste collection Odor from household waste or industrial waste generated at waste collection sites, household waste or industrial waste collection sites, waste collection sites, etc .
- Odors shoeboxes, air conditioners, cars or trucks, gas generated by cars or trucks, trains, aircraft, factories, restaurants, photo shops or photo studios, gas stations, propane gas refill stations, laundry shops or laundry factories It may be used to deodorize unpleasant odors at inns or hotels, hair salons or barber shops, auto repair shops, livestock barns, construction work sites, etc.
- the deodorant of this embodiment is isovaleric acid, acetic acid, methyl mercaptan, trimethylamine, diacetyl, acetaldehyde, formaldehyde, hydrogen sulfide, ammonia, nonenal, thioglycolic acid, thioglycolate (ammonium thioglycolate, thioglycolic acid) Monoethanolamine, sodium thioglycolate, etc.), cysteine or its derivatives, thioglycerin, sulfite, lactone thiol, odors mainly composed of cysteamine, etc., and more suitable for deodorizing tobacco odors it can.
- odors mainly composed of components selected from isovaleric acid, acetic acid, methyl mercaptan, trimethylamine, diacetyl, acetaldehyde, formaldehyde, hydrogen sulfide, ammonia and nonenal include body odors such as sweat, feet, halitosis and wrinkles ( And body odor due to aging), animal urine odor, food rancidity or rot, odor released from chemicals used in building construction materials, wallpaper, furniture, and the like.
- odors mainly composed of components selected from thioglycolic acid, thioglycolate, cysteine or derivatives thereof, thioglycerin, sulfite, lactone thiol, and cysteamine include, for example, odors of permanent agents including a reducing agent, etc. Can be mentioned.
- the deodorant of this embodiment is mainly composed of at least one component selected from the group consisting of isovaleric acid, acetic acid, methyl mercaptan, trimethylamine, diacetyl, nonenal, ammonium thioglycolate, and monoethanolamine thioglycolate. And more suitable for deodorizing tobacco odor.
- the usage pattern of the deodorant of this embodiment is not particularly limited.
- the deodorant of this embodiment can be used by spraying or coating.
- Deodorants used by spraying include aerosol spray type deodorants, mist spray type deodorants, liquid products for sprinklers, and the like.
- the mist spray type deodorant is used for, for example, domestic pet odors, toilets, kitchen garbage, cooking utensils and the like.
- the deodorant of this embodiment is added to water, and if necessary, a surfactant, ethanol, an antibacterial agent, and the like are added and filled into a mist spray bottle.
- the aerosol type deodorant is used, for example, for garbage and odors of toilets, which are strong odors in the home.
- the aerosol type deodorant is obtained, for example, by diluting the deodorant of this embodiment with water or an aqueous ethanol solution and filling the aerosol container together with a propellant (injection gas) such as LPG and carbon dioxide.
- a propellant injection gas
- the deodorant used by spraying can be mainly deodorized indoors by making it into a fine mist and scattering it in the room.
- produces in these places can also be deodorized by disperse
- Examples of the deodorant applied after application include liquid, gel, and paste deodorants.
- the liquid, gel, and paste deodorant may be in the form of an emulsion such as cream or emulsion.
- the deodorant applied and used can be used to deodorize the body odor, for example, by applying to the human body.
- the deodorant of this embodiment is a sheet-like deodorant soaked in cloth, paper, or non-woven fabric; deodorant absorbed in powder or granules; granular, pellet-like, block-like, or tablet-like gel Deodorant kneaded or adsorbed (for example, a deodorant for space described later); Deodorant adsorbed on a porous carrier such as ceramic, activated carbon, bentonite; Liquid deodorant in a container Deodorant that has a deodorizing effect by allowing a liquid, such as a container, sponge, cloth, ceramics, etc. to permeate liquid, to partially contact the deodorant in the container and vaporize the permeated deodorant; porous ceramics, etc.
- deodorant in a qualitative container, and the deodorant that has penetrated to the outside of the container is vaporized to have a deodorizing effect.
- the liquid deodorant is added directly to the malodorous source; soaks into the film or filter.
- Deodorant, or Including the deodorant in the internal or surface it is possible to use wallpaper, building materials, diapers, sanitary napkins, shoe insoles, deodorant fiber (cloth), or as a deodorant leather or the like.
- the deodorant of this embodiment may be used as a space deodorant. Since the component (active ingredient) which has a deodorizing effect volatilizes (volatilizes) gradually, the deodorizing effect for space lasts for a long time.
- the space deodorant is obtained, for example, by adsorbing or kneading the deodorant of the present embodiment on a gel or a suitable carrier.
- the space deodorant is a gelling agent (for example, carrageenan, agar, locust bean gum, polyvinyl alcohol, gum arabic, gellan gum, gelatin, carboxymethylcellulose, chitin, It can be obtained by adding to one or a combination of two or more of chitosan, sodium alginate, polyacrylamide) and solidifying.
- a gelling agent for example, carrageenan, agar, locust bean gum, polyvinyl alcohol, gum arabic, gellan gum, gelatin, carboxymethylcellulose, chitin
- the pet excrement disposal agent with a high deodorizing effect can be manufactured using the deodorizing agent of this embodiment.
- the excrement disposal agent for pet is mainly composed of bentonite, zeolite, wood powder, paper powder, etc., and sodium polyacrylate, other sodium compounds, magnesium compounds, etc. are added to this as necessary, and this embodiment Can be produced by adding an appropriate amount of water, adding an appropriate amount of water, mixing, molding and drying. By putting this in a pet toilet such as a cat and the cat excreting on the treatment agent, an excrement treatment agent having an excellent deodorizing effect can be obtained.
- the usage amount of the deodorant according to this embodiment is not particularly limited.
- the content of the deodorant of this embodiment may be, for example, 0.01 to 50% (volume / volume) based on the total amount of the mist spray type deodorant.
- the content of the deodorant of this embodiment may be 0.2 to 70% (volume / volume) based on the total amount of the aerosol type deodorant.
- the content of the deodorant of this embodiment may be, for example, 0.5 to 20% (volume / volume) based on the total amount of the space deodorant.
- an article having a deodorizing effect can be obtained by a method of mixing with other materials, molding and drying.
- the deodorizing effect can be obtained by mixing the materials other than the deodorant of the present embodiment in two or more types, molding and drying, and absorbing the deodorant of the present embodiment in the obtained molded product. Can be obtained.
- the present invention can be regarded as a method for producing a deodorant. That is, the manufacturing method of a deodorizer comprises the process of obtaining the decomposition extract of bagasse.
- the step of obtaining the bagasse decomposition extract may comprise at least one decomposition treatment step selected from the group consisting of alkali treatment, hydrothermal treatment, acid treatment, and subcritical water treatment.
- the step of obtaining the bagasse decomposition extract may further comprise a step of obtaining a fraction by passing the decomposition treatment liquid obtained by the above decomposition treatment step through a column packed with a fixed carrier. The detailed conditions of the decomposition treatment step and the step of obtaining a fraction are as described above.
- One embodiment of the present invention can also be regarded as a deodorization method including a step of applying an effective amount of a deodorant containing the above-described decomposition extract of bagasse as an active ingredient to an object to be deodorized.
- a deodorization method including a step of applying an effective amount of a deodorant containing the above-described decomposition extract of bagasse as an active ingredient to an object to be deodorized.
- one Embodiment of this invention can be regarded as the decomposition extract of bagasse for deodorizing the deodorant object.
- the aspect, usage method, usage amount, etc. of the deodorant may be the same as those described above.
- a further embodiment of the present invention can also be viewed as the use of bagasse decomposition extract for the manufacture of deodorants.
- one Embodiment of this invention can also be regarded as use of the decomposition extract of bagasse for deodorizing the deodorant object.
- the mode of the deodorant may be the same as described above.
- the anti-aging agent of the present invention has an anti-aging action.
- the anti-aging action may be an action that suppresses skin aging, and more specifically, may be an action that suppresses and / or improves skin function deterioration due to aging, ultraviolet irradiation, or the like.
- the anti-aging action may be an action that suppresses and / or improves skin wrinkles, sagging, hardening, and the like.
- Extracellular matrix components are produced from fibroblasts.
- One cause of skin aging is degradation or reduction of extracellular matrix components.
- MMP-1 matrix metalloproteinase
- elastin is degraded by elastase, which is one of extracellular matrix degrading enzymes. Elastin acts like a spring that connects collagen fibers, but elastase can also cause the skin to lose its elasticity and cause wrinkles or sagging in the skin.
- the anti-aging agent of the present invention has an action of inhibiting extracellular matrix degrading enzymes (an action of suppressing the production of extracellular matrix degrading enzymes, an action of reducing the activity of extracellular matrix degrading enzymes), for example, It has an action of inhibiting MMP-1 that degrades type I collagen and / or an action of inhibiting elastase that degrades elastin. Thereby, skin aging is suppressed. That is, it can be said that the anti-aging agent of the present invention is based on the inhibitory action of extracellular matrix degrading enzymes, and more specifically, is based on the inhibitory action of MMP-1 or elastase. It can also be said.
- the present invention can also be said to provide an extracellular matrix degrading enzyme inhibitor, and more specifically, an MMP-1 inhibitor or an elastase inhibitor.
- the action of inhibiting MMP-1 may be an action of inhibiting the production of MMP-1, or may be an action of inhibiting the activity of MMP-1.
- the action of inhibiting elastase may be an action of inhibiting the production of elastase or an action of inhibiting the activity of elastase. That is, the anti-aging agent of the present invention may be based on at least one of an MMP-1 production inhibitory action, an MMP-1 activity inhibitory action, an elastase production inhibitory action, and an elastase activity inhibitory action. .
- the anti-aging agent of the present invention may be based on an MMP-1 production inhibitory action and / or an elastase activity inhibitory action.
- the present invention can also be said to provide an MMP-1 production inhibitor, an MMP-1 activity inhibitor, an elastase production inhibitor, or an elastase activity inhibitor. It can also be said that the present invention provides an MMP-1 production inhibitor or an elastase activity inhibitor.
- the production amount of extracellular matrix components decreases due to the deterioration of fibroblasts due to aging, ultraviolet irradiation and the like.
- the fibroblasts By activating the fibroblasts, it is possible to suppress a decrease in the amount of extracellular matrix component produced.
- the anti-aging agent of the present invention also has an action of activating fibroblasts, skin aging is further suppressed. That is, it can also be said that the anti-aging agent of the present invention is based on the activation action of fibroblasts. It can also be said that the present invention provides a fibroblast activator.
- the anti-aging agent which concerns on one Embodiment contains the decomposition extract of bagasse mentioned above as an active ingredient.
- Anti-aging agents can be used as cosmetics, food compositions, pharmaceuticals or quasi drugs.
- the food composition may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like.
- the anti-aging agent may consist only of a decomposition extract of bagasse which is an active ingredient, and may further contain ingredients usable for cosmetics, food compositions, quasi drugs or pharmaceuticals.
- Materials that can be used for cosmetics, food compositions, quasi drugs or pharmaceuticals are not particularly limited, but examples include amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, and excipients.
- the content of the decomposition extract of bagasse as an active ingredient may be appropriately set according to the form of anti-aging agent, the purpose of use, etc. described later, but the anti-aging effect From the viewpoint of more effectively exhibiting the above, the following ranges are preferable on the basis of the total amount of the anti-aging agent.
- the content of the decomposition extract of bagasse is preferably 0.5% by mass or more, more preferably 1% by mass or more, still more preferably 3% by mass or more, and preferably 50% as a solid content. It is not more than mass%, more preferably not more than 40 mass%, and still more preferably not more than 30 mass%.
- the shape of the anti-aging agent is not limited, and may be any shape such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, Any dosage form such as a capsule, a troche, a liquid, or a suspension may be used.
- the cosmetic may be a lotion, a milky lotion, a lotion, a cream, a cosmetic liquid, an oil, a pack, a lip balm or other basic cosmetics, a hair styling such as a hair nick or a hair liquid, a hair restorer, It may be hair cosmetics such as hair nourishing agents, makeup cosmetics such as foundation, lipstick, blusher, eye shadow, eyeliner, mascara, eyebrow liner, and the like.
- the anti-aging agent may be administered orally or parenterally.
- the dosage is preferably such that the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, for example, 100 ⁇ g / kg (body weight). It is more preferable to administer so that it becomes the above, and it is still more preferable to administer so that it may become 150 microgram / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 150 ⁇ g / kg (body weight) or more per day, more preferably 300 ⁇ g / kg (body weight) or more, and 450 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, an anti-aging agent can be made to act on dermis with sufficient density
- the application amount to the skin is preferably such that, for example, the bagasse degradation extract is 5 ⁇ g / cm 2 or more per time. It is more preferable to apply so that it may become 10 microgram / cm ⁇ 2 > or more, and it is still more preferable to apply so that it may become 30 microgram / cm ⁇ 2 > or more. Further, the bagasse decomposition extract is preferably applied so as to be 10 ⁇ g / cm 2 or more per day, more preferably 20 ⁇ g / cm 2 or more, and more preferably 60 ⁇ g / cm 2 or more. More preferably, it is applied.
- the bagasse decomposition extract is preferably applied so as to be 500 ⁇ g / cm 2 or less per time, more preferably 400 ⁇ g / cm 2 or less, and 300 ⁇ g / cm 2 or less. More preferably, it is applied so that Further, it is preferable decomposition extract bagasse is applied such that 1 day 1000 [mu] g / cm 2 or less, more preferably are applied such that the 800 [mu] g / cm 2 or less, and 600 [mu] g / cm 2 or less More preferably, it is applied. If it is this range, an anti-aging agent can be made to act on dermis with sufficient density
- Anti-aging agents can also be used as feed and feed additives.
- the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured fish and shellfish.
- “Feed” includes anything that animals orally take for nutritional purposes. More specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Is included. Moreover, when classifying from the aspect of a feeding method, it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- extracellular matrix degrading enzyme inhibitor or the fibroblast activator may be the same as the aspect of the anti-aging agent described above. That is, the extracellular matrix degrading enzyme inhibitor or the fibroblast activator according to one embodiment is the same as the above-mentioned description of the anti-aging agent, in which “anti-aging agent” is referred to as “extracellular matrix degrading enzyme inhibitor” or “fibroblast”. It may be read as “cell activator”.
- One embodiment of the present invention provides an effective amount of an anti-aging agent, an extracellular matrix degrading enzyme inhibitor, or a fibroblast activator containing the bagasse degradation extract described above as an active ingredient. It can also be regarded as an anti-aging method, a method of inhibiting extracellular matrix degrading enzymes, or a method of activating fibroblasts. Further, one embodiment of the present invention can be regarded as a bagasse degradation extract for use in an anti-aging method, a method of inhibiting extracellular matrix degrading enzymes, or a method of activating fibroblasts.
- the subject in the above method may be a mammal, preferably a human.
- the aspect of the anti-aging agent, extracellular matrix degrading enzyme inhibitor, or fibroblast activator, administration method, dose (intake amount) and the like may be the same as those described above.
- a further embodiment of the present invention can also be viewed as the use of bagasse degradation extract for the manufacture of anti-aging agents, extracellular matrix degrading enzyme inhibitors or fibroblast activators.
- One embodiment of the present invention can also be viewed as the use of a bagasse degradation extract for anti-aging, to inhibit extracellular matrix degrading enzymes, or to activate fibroblasts.
- the aspect of the anti-aging agent, extracellular matrix degrading enzyme inhibitor, or fibroblast activator, administration method, dose (intake amount) and the like may be the same as those described above.
- the anti-glycation agent in the present specification has an anti-glycation activity, and specifically has a production-inhibiting action (glycation reaction-inhibiting action), an accumulation-inhibiting action, or a degrading action of glycation end products (AGEs). It may be a thing.
- the anti-glycation agent of the present embodiment may be, for example, a glycation end product production inhibitor (a saccharification reaction inhibitor), an accumulation inhibitor, or a degradation agent (degradation accelerator).
- the saccharification end product is a general term for products resulting from a saccharification reaction (Maillard reaction).
- Examples of the saccharification end product include CML (N ⁇ - (carboxymethyl) lysine), pentosidine, pyraline, and crossline.
- the anti-glycation agent of this embodiment may have the above-mentioned anti-glycation activity as a result of having a production-inhibiting action, an accumulation-inhibiting action or a decomposition-promoting action of a reaction intermediate in a saccharification reaction.
- Specific examples of the reaction intermediate in the saccharification reaction may include glyoxal (GO), 3-deoxyglucosone (3DG), methylglyoxal (MGO) and the like.
- the anti-glycation agent of this embodiment contains the above-described decomposition extract of bagasse as an active ingredient.
- the content of the decomposition extract of bagasse in the anti-glycation agent may be 0.01 to 100% by mass or 0.1 to 100% by mass based on the total amount of the anti-glycation agent.
- the anti-glycation agent of this embodiment may contain excipients in addition to the bagasse decomposition extract.
- excipients when the anti-glycation agent is for animals, various starches such as corn starch and wheat starch, dextrin, various gluten, various rice brans such as wheat flour, bran, defatted rice bran, soybeans such as soybean meal, yellow flour,
- sugars such as glucose and lactose
- oils and fats such as vegetable oil and animal oil, fish meal, yeasts, silicon compounds
- various phosphates diatomaceous earth, bentonite and other minerals
- feed and feed additives Excipients that can be used in In addition, examples of the excipient include sugars such as lactose, starch, maltose, and other excipients that can be used for producing human preparations when the anti-glycation agent is for humans.
- corn starch, dextrin and defatted rice bran can be used as a carrier for pharmaceutical preparations.
- an anti-glycation agent can be used, for example, in the form of powder, granules, or tablets. It can be set as a solid preparation.
- the anti-glycation agent of this embodiment may exhibit an anti-glycation effect by administration (oral administration or parenteral administration) to humans or animals.
- the dosage is, for example, preferably administered such that the bagasse degradation extract is 100 ⁇ g or more per kg body weight, and is administered so that it is 150 ⁇ g or more. It is more preferable that the dose is 200 ⁇ g or more.
- the bagasse degradation extract is preferably administered so as to be 200 ⁇ g or more per day per kg of body weight, more preferably 300 ⁇ g or more, and more preferably 400 ⁇ g or more. Is more preferable.
- the bagasse decomposition extract is preferably administered to 2000 mg or less per kg body weight, more preferably 1500 mg or less, more preferably 1000 mg or less. More preferably.
- the bagasse degradation extract is preferably administered at a dose of 4000 mg or less per kg body weight per day, more preferably at a dose of 3000 mg or less, and a dose of 2000 mg or less. Is more preferable. If it is this range, sufficient blood concentration can be achieved and anti-glycation activity can be expressed better.
- the dose of the anti-glycation agent can be appropriately determined depending on the degree of purification of the bagasse degradation extract, the form, the type of the target animal, the health condition, the degree of growth, and the like.
- the mode of administration such as intensive or long-term administration, is an important factor in determining the dosage.
- the dose of the anti-glycation agent may be 50 to 3000 mg or 100 to 2000 mg per kg body weight per day based on the total amount (solid content) of the bagasse degradation extract.
- the administration period in the case of intensive administration may be 1 to 20 days.
- the dose of the anti-glycation agent may be 1 to 500 mg or 1 to 100 mg per day per kg of body weight based on the total amount (solid content) of the bagasse degradation extract.
- the administration period in the case of long-term administration may be, for example, several weeks to several months (for example, 20 to 180 days). If it is this range, sufficient blood concentration can be achieved and anti-glycation activity can be expressed better.
- the anti-glycation agent of the present embodiment can be used as a component of products such as pharmaceuticals, quasi drugs, food and drink (food composition), feed, feed additives and the like.
- foods and drinks include health foods, functional display foods, special-purpose foods, nutritional supplements, supplements, and foods for specified health use.
- the said anti-glycation agent can also be used as a component in foodstuffs, such as seasonings (soy sauce, miso, etc.), confectionery, or drinks, such as water, soft drinks, fruit juice drinks, and alcoholic drinks.
- feed examples include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured seafood.
- feed includes anything that animals orally take for nutritional purposes. Specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Includes. Moreover, when classifying from the aspect of a feeding method, it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- the product for example, a food or drink
- the product comprising or containing the anti-glycation agent of the present embodiment may be for anti-glycation. That is, the food / beverage products containing the anti-glycation agent of this embodiment can be used suitably as an anti-glycation food / beverage product or an anti-glycation food composition.
- the shape of the product containing the anti-glycation agent may be either solid or liquid.
- the content of the anti-glycation agent contained in the product may be appropriately determined according to the type of the product and the intake method. From the viewpoint of more effectively exhibiting the anti-glycation effect, the product preferably contains a decomposition extract of bagasse in an amount such that the solid content is 0.001% by mass or more.
- the intake amount of the product is preferably 50 to 3000 mg based on the total amount (solid content) of the decomposition extract of bagasse / Kg (body weight), more preferably 100 to 2000 mg / kg (body weight).
- the above-mentioned product intake is preferably 1 to 500 mg / kg (body weight) based on the total amount (solid content) of the bagasse degradation extract.
- One embodiment of the present invention can also be regarded as an anti-glycation method including a step of administering an effective amount of an anti-glycation agent containing the above-described decomposition extract of bagasse as an active ingredient to a subject in need thereof. .
- one Embodiment of this invention can be regarded as the decomposition extract of bagasse for using for an anti-glycation method.
- the subject in the above method may be a mammal, preferably a human.
- the aspect of the anti-glycation agent, administration method, dose (intake amount), etc. may be the same as those described above.
- a further embodiment of the present invention can also be viewed as the use of bagasse degradation extract for the production of anti-glycation agents. Further, one embodiment of the present invention can be regarded as the use of a bagasse decomposition extract for anti-glycation.
- the aspect of the anti-glycation agent, administration method, dose (intake amount), etc. may be the same as those described above.
- the anti-type I allergy agent in this specification is a composition provided with the effect
- the action of suppressing type I allergic symptoms may be an action of alleviating, treating, or preventing symptoms due to type I allergies such as hay fever, urticaria, allergic rhinitis, bronchial asthma and the like.
- the action of suppressing type I allergic symptoms may be an action of suppressing degranulation of mast cells or basophils in the mechanism of type I allergic reaction. That is, the anti-type I allergic agent in the present specification may be a mast cell or basophil degranulation inhibitor.
- the anti-type I allergic agent which concerns on one Embodiment contains the decomposition extract of bagasse as an active ingredient.
- the anti-type I allergic agent according to the present embodiment may consist only of a decomposition extract of bagasse which is an active ingredient, and may further contain a material that can be used for food, quasi drugs or pharmaceuticals.
- Materials that can be used for foods, quasi drugs or pharmaceuticals are not particularly limited. For example, amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients, binders , Lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- the content of the decomposition extract of bagasse as an active ingredient may be appropriately set according to the form, purpose of use, etc. of the anti-type I allergic agent described later.
- it is preferably 100 ⁇ g / g or more, more preferably 25 ⁇ g / g or more, still more preferably 400 ⁇ g / g, based on the total amount of the anti-I allergic agent.
- it is preferably 10 mg / g or less, more preferably 7.5 mg / g or less, and still more preferably 5 mg / g or less.
- the anti-type I allergic agent may be in any form such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, capsule, Any dosage form such as a troche, liquid or suspension may be used.
- the anti-type I allergic agent of the present embodiment releases granules containing chemical mediators such as histamine and leukotriene from the mast cells or basophils in particular (degranulation). Has the action of suppressing degranulation (the action of suppressing degranulation). Therefore, according to the anti-type I allergic agent of this embodiment, the symptoms caused by the type I allergic reaction can be effectively suppressed, treated or prevented.
- an anti-type I allergic agent has a degranulation inhibitory effect is determined, for example, by the fact that IgE bound to the cell surface such as rat basophilic leukemia cells (RBL-2H3 cells) is cross-linked by an antigen.
- RBL-2H3 cells rat basophilic leukemia cells
- an anti-type I allergic agent is added compared to a sample to which no anti-type I allergic agent is added It can be confirmed by calculating how much the degranulation of the specimen was suppressed.
- Anti-type I allergic agents can be used as foods, quasi drugs or pharmaceuticals.
- the food may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like.
- the anti-type I allergic agent may be administered parenterally such as intravenous administration, or may be administered orally.
- the anti-type I allergic agent is preferably administered orally.
- the dosage is preferably such that the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, for example, 150 ⁇ g / kg. It is more preferable to administer so that it may become (body weight) or more, and it is still more preferable to administer so that it may become 250 microgram / kg (body weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 100 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 500 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and an anti-type I allergic effect can be expressed better.
- the dose is preferably administered so that the decomposition extract of bagasse is 50 ⁇ g / kg (body weight) or more, and 100 ⁇ g / kg (body weight). It is more preferable to administer so that it becomes the above, and it is still more preferable to administer so that it may become 150 microgram / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 150 ⁇ g / kg (body weight) or more per day, more preferably 300 ⁇ g / kg (body weight) or more, and 450 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and an anti-type I allergic effect can be expressed better.
- the anti-type I allergic agent of the present embodiment has the above-described action, it can be used for patients who have symptoms of type I allergy and for those who have not developed allergy who want to prevent type I allergy in advance. .
- the specific aspect of the degranulation inhibitor of mast cells or basophils according to one embodiment may be the same as the aspect of the anti-type I allergy agent described above. That is, the degranulation inhibitor of mast cells or basophils according to one embodiment is the “anti-type I allergic agent” in the above description of the anti-type I allergy agent. It may be read as “agent”.
- One embodiment of the present invention provides an effective amount of an anti-type I allergic agent containing the above-mentioned bagasse degradation extract as an active ingredient, or an inhibitor of degranulation of mast cells or basophils. It can also be regarded as an anti-type I allergy method or a method for suppressing degranulation of mast cells or basophils. Further, one embodiment of the present invention can be regarded as a bagasse degradation extract for use in an anti-type I allergy method or a method of suppressing degranulation of mast cells or basophils.
- the subject in the above method may be a mammal, preferably a human.
- the aspect, administration method, dose (intake amount), etc. of the anti-type I allergic agent or mast cell or basophil degranulation inhibitor may be the same as those described above.
- a further embodiment of the present invention can be regarded as the use of a bagasse degradation extract for the production of an anti-type I allergic agent or a mast cell or basophil degranulation inhibitor.
- One embodiment of the invention can also be viewed as the use of a bagasse degradation extract for anti-type I allergy or to suppress degranulation of mast cells or basophils.
- the mode of the anti-type I allergic agent or the mast cell or basophil degranulation inhibitor may be the same as described above.
- the antihypertensive agent of the present invention has an antihypertensive action.
- the antihypertensive effect may be an effect of suppressing an increase in blood pressure.
- Angiotensin II has an action of constricting blood vessels and an action of increasing blood volume by suppressing discharge of sodium or water in the kidney, and has a function of increasing blood pressure.
- Angiotensin II is produced when angiotensin I is converted by an angiotensin converting enzyme (ACE). For this reason, an increase in blood pressure can be suppressed by inhibiting ACE and suppressing the production of angiotensin II.
- ACE angiotensin converting enzyme
- the antihypertensive agent of the present invention has an action of inhibiting ACE, the production of angiotensin II is suppressed, and as a result, an increase in blood pressure is suppressed. That is, the antihypertensive agent of the present invention can be said to be based on the inhibitory action of angiotensin converting enzyme, and can also be said to be based on the action of suppressing the production of angiotensin II. It can also be said that the present invention provides an angiotensin converting enzyme inhibitor.
- the antihypertensive agent according to one embodiment contains the above-described decomposition extract of bagasse as an active ingredient.
- Antihypertensive agents can be used as food compositions, pharmaceuticals or quasi drugs.
- the food composition may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like.
- the antihypertensive agent may consist only of a decomposition extract of bagasse which is an active ingredient, and may further contain a material usable for a food composition, quasi-drug or pharmaceutical.
- the material that can be used for the food composition, quasi-drug, or pharmaceutical is not particularly limited.
- amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients examples include binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- the content of the decomposition extract of bagasse which is an active ingredient, may be appropriately set according to the form of antihypertensive agent described below, the purpose of use, etc. Is more preferably 1% by mass or more, more preferably 3% by mass or more, still more preferably 5% by mass or more, and preferably 50% by mass from the viewpoint of more effectively exhibiting It is below, More preferably, it is 40 mass% or less, More preferably, it is 30 mass% or less.
- the shape of the antihypertensive agent is not limited and may be any shape such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, Any dosage form such as a capsule, a troche, a liquid, or a suspension may be used.
- the antihypertensive agent may be administered orally or parenterally such as intravenous administration.
- the dose is preferably administered so that the decomposition extract of bagasse is 50 ⁇ g / kg (body weight) or more per dose, for example, 100 ⁇ g / kg (body weight). It is more preferable to administer so that it becomes the above, and it is still more preferable to administer so that it may become 150 microgram / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 150 ⁇ g / kg (body weight) or more per day, more preferably 300 ⁇ g / kg (body weight) or more, and 450 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and an antihypertensive action can be expressed better.
- the dosage is preferably such that the bagasse degradation extract is 100 ⁇ g / kg (body weight) or more, and 150 ⁇ g / kg (body weight). ) Is more preferably administered, and even more preferably 200 ⁇ g / kg (body weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 200 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 400 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and an antihypertensive action can be expressed better.
- Anti-hypertensive agents can also be used as feed and feed additives.
- the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured fish and shellfish.
- “Feed” includes anything that animals orally take for nutritional purposes. More specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Is included. Moreover, when classifying from the aspect of a feeding method, it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- angiotensin converting enzyme inhibitor may be the same as the aspect of the antihypertensive agent described above. That is, the angiotensin converting enzyme inhibitor according to one embodiment may be obtained by replacing “antihypertensive agent” with “angiotensin converting enzyme inhibitor” in the above description of the antihypertensive agent.
- One embodiment of the present invention comprises the step of administering to a subject in need thereof an effective amount of an antihypertensive agent or an angiotensin converting enzyme inhibitor containing the bagasse degradation extract described above as an active ingredient. It can also be regarded as a hypertension method or an angiotensin converting enzyme inhibition method. Moreover, one Embodiment of this invention can be regarded as the decomposition extract of bagasse for using for the antihypertensive method or the inhibition method of an angiotensin converting enzyme.
- the subject in the above method may be a mammal, preferably a human.
- the aspect of an antihypertensive agent or an angiotensin converting enzyme inhibitor, administration method, dose (intake amount), etc. may be the same as those described above.
- a further embodiment of the present invention can be regarded as the use of a bagasse degradation extract for the production of an antihypertensive agent or an angiotensin converting enzyme inhibitor.
- One embodiment of the present invention can also be viewed as the use of a bagasse degradation extract for anti-hypertension or to inhibit angiotensin converting enzyme.
- the aspect of the antihypertensive agent or the angiotensin converting enzyme inhibitor may be the same as described above.
- flavor improver> The flavor improvement agent of this invention has the effect
- flavors are flavors that are perceived as desirable by humans, and examples include deliciousness, refreshing feeling, refreshing feeling, ease of eating, ease of drinking, mellowness, mouthfeel, and the original flavor of the material.
- Unpleasant taste refers to a flavor that people feel unfavorable, such as acidity, bitterness, odor, egg taste, miscellaneous taste, disgusting aftertaste, deterioration odor, retort sterilization odor, dry odor, oily odor, egg Examples include odor, animal odor of meat, raw odor of fish, blue odor of beans and vegetables, oily feeling, irritation, powderiness, stickiness of taste, metal taste and the like.
- the flavor improver of the present invention has an effect of enhancing the preferred flavor of food and drink.
- action which strengthens the preferable flavor of food / beverage products may be that the preferable flavor mentioned above comes to be felt more strongly. That is, the flavor improving agent according to one embodiment may be based on the action of enhancing the preferred flavor of food and drink. Moreover, it can also be said that this invention provides the preferable flavor enhancer of food-drinks.
- the flavor improver of the present invention has an effect of reducing the taste of food and drink.
- action which reduces the unpleasant taste of food / beverage products may be that the unpleasant taste of food / beverage products mentioned above becomes difficult to feel. That is, the flavor improving agent according to one embodiment may be based on the action of reducing the taste of food and drink. Moreover, it can also be said that this invention provides the taste reduction agent of food-drinks.
- the flavor improver may have one of the actions of enhancing the preferred flavor of the food and drink and the action of reducing the taste, or may have both actions.
- the flavor improver according to one embodiment contains the bagasse decomposition extract described above.
- the flavor enhancer may be composed only of a bagasse decomposition extract, and may further be blended with a material that can be used in a food composition (food or beverage).
- a material that can be used in a food composition is not particularly limited.
- amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, fragrances, excipients, binders, lubricants, Disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like can be mentioned.
- the content of the bagasse decomposition extract may be set as appropriate according to the purpose of use of the flavor enhancer, etc. Therefore, the solid content is preferably 0.1% by mass or more, more preferably 0.3% by mass or more, still more preferably 0.5% by mass or more, and preferably 90% by mass or less. Yes, more preferably 80% by mass or less, still more preferably 70% by mass or less.
- the flavor enhancer described above can improve the flavor of food and drink by adding it to various foods and drinks. More specifically, by adding a flavor improver to a food or drink, the preferred flavor of the food or drink can be enhanced, and / or the taste and taste of the food or drink can be reduced.
- Adding a flavor enhancer to a food or drink includes attaching or impregnating the flavor enhancer to the food or drink.
- food-drinks contain a flavor improvement agent. That is, the food / beverage products which concern on one Embodiment are the food / beverage products which contained the flavor improvement agent containing the decomposition extract of bagasse, and the flavor improved.
- the food / beverage products containing the flavor improver include food / beverage products to which the flavor improver is attached and food / beverage products impregnated with the flavor improver.
- Food and drink includes general food and drink, health food, food for specified health use, functional food, functional nutrition food, supplement and the like.
- Foods and beverages include milk drinks, lactic acid bacteria drinks, soy milk drinks, vegetable drinks, fruit drinks, teas, coffee drinks, alcoholic drinks, and other soft drinks (such as drinks containing vinegar); noodles, breads, Food products such as processed vegetable products, processed fruit products, meat products, processed seafood, dairy products, legume preparations, soups, and seasonings may be used.
- the food / beverage product with which the flavor improving agent can improve the flavor is preferably a soy milk beverage, a beverage containing vinegar, a lactic acid bacteria beverage, a meat product, or processed seafood.
- the flavor improver can enhance the preferred flavor of the soy milk drink.
- the preferred flavor of the soy milk drink may be a refreshing feeling, a delicious taste, a refreshing feeling, ease of drinking, and the like.
- the flavor improver can reduce the taste of soy milk beverages.
- the unpleasant taste of the soy milk beverage may be the blue odor of the beans, the unpleasant aftertaste, the egy taste, the miscellaneous taste and the like.
- the flavor improver can enhance the preferred flavor of the vinegar-containing beverage.
- the preferred flavor of the vinegar-containing beverage may be ease of drinking, taste, refreshing feeling, mellowness, mouthfeel and the like.
- the flavor improver can reduce the taste of vinegar-containing beverages.
- the taste of vinegar-containing beverages may be sour, unpleasant aftertaste, and the like.
- the flavor improver can enhance the preferred flavor of the lactic acid bacteria beverage.
- the preferred flavor of the lactic acid bacteria beverage may be a refreshing feeling, a refreshing feeling, ease of drinking, deliciousness, sharpness of aftertaste, and the like.
- the flavor improver can reduce the taste of lactic acid bacteria beverages.
- the unpleasant taste of the lactic acid bacteria beverage may be a nasty taste or the like.
- the flavor improver can enhance the preferred flavor of the meat product.
- the preferred flavor of the meat product may be the original flavor, taste, ease of eating, freshness, etc. of the meat.
- the flavor enhancer can reduce the taste of meat products.
- the taste of meat products may be a bad aftertaste, meat odor, oiliness, and the like.
- the flavor improver can enhance the preferred flavor of the processed seafood.
- the preferred flavor of processed seafood may be the original flavor, taste, ease of eating, freshness, etc. of seafood.
- the flavor improver can reduce the taste of processed seafood.
- the taste of processed seafood may be an unpleasant aftertaste, seafood odor, oiliness, and the like.
- the amount of the flavor enhancer added to the food or drink can be appropriately selected depending on the kind of the food or drink to be added.
- the addition amount of the decomposition extract of bagasse contained in the flavor improver is 0.3 mass ppm or more, 0.6 mass ppm or more, 0.8 mass ppm or more, 3 mass ppm or more, based on the total amount of food and drink. It may be not less than ppm by mass, or not less than 10 ppm by mass, and may be not more than 50 ppm by mass, not more than 40 ppm by mass, and not more than 30 ppm by mass. If it is the addition amount in this range, a sufficient flavor improving effect can be obtained.
- Specific embodiments of the preferred flavor enhancer for foods and drinks and the taste-reducing agent for foods and beverages according to one embodiment may be the same as those in the above-described flavor improvers. That is, the preferred flavor enhancer for foods and beverages according to one embodiment and the taste-reducing agent for foods and beverages are the “flavor enhancers for foods and beverages” in the description of the flavor improvers described above, or It may be read as “a taste-reducing agent for food and drink”.
- One embodiment of the present invention requires an effective amount of a flavor enhancer, a preferred flavor enhancer for foods and beverages, or a taste-reducing agent for foods and beverages containing the bagasse decomposition extract described above as an active ingredient. It can also be regarded as a flavor improving method including a step of adding to the subject, a method for enhancing a preferable flavor of food or drink, or a method for reducing a taste of food or drink. Moreover, one Embodiment of this invention can be regarded as the decomposition extract of bagasse for using for the flavor improvement method, the enhancement method of the preferable flavor of food / beverage products, or the taste reduction method of food / beverage products.
- the object in the above method may be a food or drink.
- the aspect of the flavor enhancer, the preferred flavor enhancer for foods and drinks, or the taste reduction agent for foods and drinks, the administration method, the dose (intake amount), and the like may be the same as those described above.
- a further embodiment of the present invention can also be regarded as the use of a bagasse decomposition extract for the production of a flavor enhancer, a preferred flavor enhancer for foods and beverages, or a taste reduction agent for foods and beverages.
- one embodiment of the present invention may be regarded as the use of a decomposition extract of bagasse for improving the flavor of food or drink, enhancing the preferred flavor of food or drink, or reducing the taste of food or drink. it can.
- the aspect of the flavor enhancer, the preferred flavor enhancer for foods and drinks, or the taste reduction agent for foods and drinks may be the same as described above.
- the muscle enhancing agent of the present invention has a muscle enhancing action.
- the muscle enhancing action in the present specification includes a myotube differentiation promoting action that promotes differentiation from myoblasts to myotube cells, and a mitochondrial activating action that activates mitochondria in muscle. That is, it can be said that the present invention provides a myotube differentiation promoter or a mitochondrial activator.
- the mitochondrial activation action is an action that increases the amount of mitochondria present in cells (muscle cells) in muscle (the amount of mitochondria per cell). Furthermore, the activity of mitochondria present in muscle cells (mitochondrial activity per cell) ) Is included. That is, the present invention can also be said to provide a mitochondrial extender in muscle cells or a mitochondrial activator in muscle cells.
- the muscle strengthening agent which concerns on one Embodiment contains the decomposition extract of bagasse mentioned above as an active ingredient.
- the muscle enhancer can be used as a food composition, a pharmaceutical product or a quasi drug.
- the food composition may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like. That is, according to the present invention, it can be said that a food composition for enhancing muscles, a pharmaceutical product for enhancing muscles, or a quasi-drug for enhancing muscles is provided.
- the muscle potentiator may consist only of the decomposed extract of bagasse, which is an active ingredient, and may further contain materials that can be used for food compositions, quasi drugs or pharmaceuticals.
- the material that can be used for the food composition, quasi-drug, or pharmaceutical is not particularly limited.
- amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients examples include binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- the content of the decomposition extract of bagasse as an active ingredient may be appropriately set according to the form and purpose of use of the muscle strengthening agent described later. Is more preferably 0.5% by mass or more, more preferably 1% by mass or more, still more preferably 3% by mass or more, and preferably 50% or more from the viewpoint of more effectively exhibiting It is not more than mass%, more preferably not more than 40 mass%, and still more preferably not more than 30 mass%.
- the shape of the muscle enhancer is not limited, and may be any shape such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet, Any dosage form such as a capsule, a troche, a liquid, or a suspension may be used.
- the muscle enhancer may be administered orally or parenterally such as intravenous administration.
- the dose is preferably administered such that the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, for example, 100 ⁇ g / kg (body weight). It is more preferable to administer so that it becomes the above, and it is still more preferable to administer so that it may become 150 microgram / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 150 ⁇ g / kg (body weight) or more per day, more preferably 300 ⁇ g / kg (body weight) or more, and 450 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and a muscle reinforcement
- the dosage is preferably such that, for example, the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, and 150 ⁇ g / kg ( (Body weight) is more preferably administered, more preferably 250 ⁇ g / kg (body weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 100 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 500 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and a muscle reinforcement
- the muscle enhancer of this embodiment can be used for humans or animals.
- the muscle enhancing agent When used for animals, it can be used as a feed or a feed additive.
- the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured fish and shellfish.
- “Feed” includes anything that animals orally take for nutritional purposes. More specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Is included.
- when classifying from the aspect of a feeding method it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- the muscle enhancer according to the present embodiment may be used to promote muscle formation, promote recovery of damaged muscle, or activate the function of the formed muscle.
- mitochondrial activator or myotube differentiation promoter may be the same as the aspect of the muscle enhancer described above. That is, the mitochondrial activator or myotube cell differentiation promoting agent according to one embodiment is the “muscle enhancing agent” as “mitochondrial activator” or “myotube cell differentiation promoting agent” in the above description of the muscle potentiator. It may be replaced.
- an effective amount of a muscle potentiator, mitochondrial activator, or myotube differentiation promoter containing the above-described bagasse degradation extract as an active ingredient is administered to a subject in need thereof. It can also be regarded as a muscle enhancement method, a mitochondrial activation method, or a myotube differentiation promotion method including steps. Further, one embodiment of the present invention can be regarded as a bagasse degradation extract for use in a muscle enhancement method, a mitochondrial activation method, or a differentiation promotion method into myotube cells.
- the subject in the above method may be a mammal, preferably a human.
- the aspect, administration method, dose (intake amount) and the like of the muscle enhancer, mitochondrial activator, or myotube differentiation promoter may be the same as those described above.
- a further embodiment of the present invention can be regarded as the use of a bagasse degradation extract for the production of a muscle potentiator, a mitochondrial activator, or a myotube differentiation promoter.
- One embodiment of the invention can also be viewed as the use of bagasse degradation extract to enhance muscle, activate mitochondria, or promote differentiation into myotube cells.
- the aspect of the muscle enhancer, mitochondrial activator, or myotube differentiation promoter may be the same as described above.
- the bone metabolism improving agent of the present invention has an effect of improving bone metabolism.
- the effect of improving bone metabolism may be at least one of an effect of promoting bone formation (new bone formation) and an effect of suppressing excessive bone resorption (bone destruction).
- the balance between bone formation and bone resorption can be adjusted suitably, and as a result, bone reconstruction can be facilitated. That is, the present invention can be said to provide an osteogenesis promoter and a bone resorption inhibitor, and it can also be said to provide a balance regulator of bone formation and bone resorption.
- the promotion of bone formation in the osteogenesis promoter may be based on the action of promoting osteoblast differentiation. That is, the osteogenesis promoter in this specification can also be referred to as an osteoblast differentiation promoter.
- the suppression of bone resorption in the bone resorption inhibitor may be based on the action of suppressing osteoclast differentiation. That is, the bone resorption inhibitor in this specification can also be called osteoclast differentiation inhibitor.
- the osteogenesis promoter according to an embodiment contains the above-described decomposition extract of bagasse as an active ingredient.
- the bone metabolism improving agent can be used as a food composition, a pharmaceutical product or a quasi drug.
- the food composition may be provided in the form of, for example, a health food, a food for specified health use, a functional food, a functional nutrition food, a supplement, and the like. That is, according to the present invention, it can also be said that a food composition for improving bone metabolism, a pharmaceutical product for improving bone metabolism, or a quasi-drug for improving bone metabolism is provided.
- the bone metabolism-improving agent may consist only of a decomposition extract of bagasse, which is an active ingredient, and may further contain a material that can be used for food compositions, quasi drugs, or pharmaceuticals.
- the material that can be used for the food composition, quasi-drug, or pharmaceutical is not particularly limited.
- amino acids, proteins, carbohydrates, fats and oils, sweeteners, minerals, vitamins, flavors, excipients examples include binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents and the like.
- the shape of the bone metabolism improving agent is not limited and may be any shape such as solid (powder, granule, etc.), liquid (solution, suspension, etc.), paste, etc., powder, pill, granule, tablet , Any dosage form such as capsules, troches, solutions, suspensions and the like.
- the bone metabolism improving agent may be administered orally or parenterally such as intravenous administration.
- the dose is preferably administered such that the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more per dose, and 100 ⁇ g / kg (body weight). ) Is more preferably administered, more preferably 150 ⁇ g / kg (body weight) or more.
- the bagasse decomposition extract is preferably administered at 150 ⁇ g / kg (body weight) or more per day, more preferably 300 ⁇ g / kg (body weight) or more, and 450 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered at a dose of 1000 mg / kg (body weight) or less, more preferably 800 mg / kg (body weight) or less, more preferably 600 mg / kg. More preferably, it is administered so as to be not more than kg (body weight).
- the bagasse degradation extract is preferably administered at a dose of 3000 mg / kg (body weight) or less per day, more preferably at a dose of 2000 mg / kg (body weight) or less, and 1000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and a bone metabolism improvement effect can be expressed more effectively.
- the dose is preferably administered such that, for example, the bagasse degradation extract is 50 ⁇ g / kg (body weight) or more at a time, and 150 ⁇ g / kg. It is more preferable to administer so that it may become (body weight) or more, and it is still more preferable to administer so that it may become 250 microgram / kg (body weight) or more.
- the bagasse degradation extract is preferably administered at a daily dose of 100 ⁇ g / kg (body weight) or more, more preferably 300 ⁇ g / kg (body weight) or more, and 500 ⁇ g / kg. It is more preferable to administer it so that it becomes (weight) or more.
- the bagasse decomposition extract is preferably administered so as to be 2000 mg / kg (body weight) or less, more preferably 1500 mg / kg (body weight) or less, more preferably 1000 mg. More preferably, it is administered so that it is not more than / kg (body weight).
- the bagasse decomposition extract is preferably administered so as to be 4000 mg / kg (body weight) or less per day, more preferably 3000 mg / kg (body weight) or less, more preferably 2000 mg / kg. It is more preferable to administer it so that it is (weight) or less. If it is this range, sufficient blood concentration can be achieved and a bone metabolism improvement effect can be expressed more effectively.
- the bone metabolism improving agent of this embodiment can be used for humans or animals.
- the bone metabolism improving agent when used for animals, it can be used as a feed or a feed additive.
- the feed include feed for companion animals such as dog food and cat food, feed for livestock, feed for poultry, and feed for cultured fish and shellfish.
- “Feed” includes anything that animals orally take for nutritional purposes. More specifically, when classified from the aspect of nutrient content, it includes all of roughage, concentrated feed, inorganic feed, and special feed, and from the aspect of official standards, it includes all of mixed feed, mixed feed, and simple feed. Is included.
- when classifying from the aspect of a feeding method it includes all feeds that are fed directly, feeds that are mixed with other feeds, or feeds that are added to drinking water to replenish nutrients.
- the above-mentioned bone metabolism improving agent can improve bone metabolism of humans or non-human animals, it can also be used for the prevention and treatment of bone-related diseases such as fractures, osteoporosis and osteomalacia. it can.
- the bone formation promoter and the bone resorption inhibitor according to one embodiment may be the same as those in the bone metabolism improving agent described above. That is, the bone formation promoter or bone resorption inhibitor according to one embodiment is the above-mentioned description of the bone metabolism improving agent, the “bone metabolism improving agent” as “bone formation promoting agent” or “bone resorption inhibitor”. It may be replaced.
- an effective amount of a bone metabolism improving agent, osteogenesis promoting agent, or bone resorption inhibitor containing the above-described decomposition extract of bagasse as an active ingredient is administered to a subject in need thereof. It can also be regarded as a bone metabolism improvement method, a bone formation promotion method, or a bone resorption suppression method including steps. In addition, one embodiment of the present invention can be regarded as a bagasse decomposition extract for use in a bone metabolism improvement method, a bone formation promotion method, or a bone resorption suppression method.
- the subject in the above method may be a mammal, preferably a human.
- the aspect, administration method, dose (intake amount), and the like of the bone metabolism improving agent, bone formation promoter, or bone resorption inhibitor may be the same as those described above.
- a further embodiment of the present invention may be regarded as the use of a bagasse degradation extract for the production of a bone metabolism improving agent, a bone formation promoter, or a bone resorption inhibitor.
- One embodiment of the present invention can also be viewed as the use of a bagasse degradation extract to improve bone metabolism, promote bone formation, or inhibit bone resorption.
- the aspect of the bone metabolism improving agent, osteogenesis promoter, or bone resorption inhibitor may be the same as described above.
- the decomposition extract of bagasse may be simply expressed as “extract”.
- Extract A The fraction eluted from the resin tower was concentrated under reduced pressure to a concentration of about 10 times using a rotary evaporator, and then freeze-dried overnight to obtain 20 g of a brown powder as a decomposition extract of bagasse. This was designated as Extract A.
- Extract W The fraction eluted from the resin tower was concentrated under reduced pressure to a concentration of about 10 times using a rotary evaporator, and then freeze-dried overnight to obtain 40 g of brown powder as a decomposition extract of bagasse. This was designated as Extract W.
- Extract A was dissolved in water to prepare a 50 mg / mL test solution stock solution. This test solution stock solution was diluted with a DMEM medium to prepare a test solution having a specimen concentration of 2000 ⁇ g / mL.
- [3T3-L1 cell fat accumulation suppression test] (Cell culture) Mouse fat precursor cells 3T3-L1 cells (National Institute of Biomedical Innovation, Health and Nutrition) are seeded in a 24-well plate, and calf serum (10 vol%, based on the total amount of medium) and penicillin-streptomycin solution (1 vol) %, Based on the total amount of medium). After culturing, the culture medium was replaced with DMEM medium containing fetal bovine serum (10 vol%, medium total volume standard) and penicillin-streptomycin solution (1 vol%, medium total volume standard), and 3T3- using Adipogenesis Assay Kit (Cayman Chemical Co.). L1 cells were differentiated into adipocytes.
- the prepared 2000 ⁇ g / mL test solution was added so that the final concentration was 1000 ⁇ g / mL (Example a1).
- the medium was replaced with a newly prepared test solution-added medium.
- progenitor cells undifferentiated a1-1
- cells to which no test solution was added were untreated controls (Comparative Example a1-1)
- berberine chloride Wako Pure Chemical Industries, Ltd.
- the same test was conducted as a positive control (positive control a1-1) to which a final concentration of 1 ⁇ g / mL was added. These were further cultured for 4 days.
- Cell staining After culturing, the culture supernatant was removed, the cells were fixed according to the procedure of Adipogenesis Assay Kit, and then the cells were stained with an oil red O solution which is a lipophilic dye for staining lipid droplets.
- FIG. 1 (a) is a progenitor cell (undifferentiated a1-1), (b) is a comparative example a1-1, (c) is a positive control a1-1, and (d) is an observation result of Example a1. .
- a progenitor cell undifferentiated a1-1
- b is a comparative example a1-1
- c is a positive control a1-1
- d is an observation result of Example a1.
- accumulation of fat droplets stained with oil red O was observed in the fat cells of Comparative Example a1-1, but the fat cells of Example a1 were compared with the fat cells of Comparative Example a1-1. The accumulated fat droplets were few.
- Fat accumulation rate (%) ⁇ (Sa ⁇ BL Avr ) / (CN ⁇ BL Avr ) Avr ⁇ ⁇ 100
- the results of calculating the fat accumulation rate are shown in FIG.
- the fat accumulation rate was 100 ⁇ 5.5% for Comparative Example a1-1, while 19 ⁇ 1.0 for Positive Control a1-1 and 15 ⁇ 1.5 for Example a1.
- Extract W was dissolved in ethanol to prepare a 50 mg / mL test solution stock solution. This test solution stock solution was diluted with a DMEM medium to prepare test solutions having a specimen concentration of 500 ⁇ g / mL.
- ⁇ Test b Test as an antidementia agent> [Preparation of test solution]
- the extract A and the extract W were stored in a powder state at room temperature (control temperature: 18.0 to 28.0 ° C.) until the test.
- Water for injection (Otsuka distilled water, Otsuka Pharmaceutical Factory Co., Ltd.) was prepared as a medium for dissolving the extract. The required amount of the extract was weighed and dissolved in water for injection, and then diluted to a predetermined concentration, which was used as a test solution.
- Amyloid ⁇ (Amyloid- ⁇ Protein (25-35), manufactured by Polypeptide Laboratories) used for the amyloid ⁇ solution is frozen (control temperature: ⁇ 30 ° C. to ⁇ 20 ° C. (actual value: ⁇ 27.1) until it is used for the test.
- the amyloid ⁇ was dissolved in water for injection to 2 mM to prepare an amyloid ⁇ solution.
- mice Male Slc: ddY mice (SPF, manufactured by Nippon SLC Co., Ltd.) were used as test animals. As the mouse, a 5-week-old mouse was obtained. The mouse is an animal species generally used in behavioral pharmacology tests, and its lineage maintenance is clear. The body weight range of mice one day after acquisition was 23.8 to 30.0 g. The obtained mice were provided with a 5-day preliminary breeding period.
- mice have a management temperature of 20.0 to 26.0 ° C, a management humidity of 40.0 to 70.0%, light and darkness for 12 hours each (lighting: 6 am to 6 pm), and ventilation rate 12 times / hour (with a filter) Breeding in an animal breeding room maintained in fresh air. From the pre-breeding period to the grouping date, grouping up to 10 animals per cage using plastic cages (W: 310 x D: 360 x H: 175 mm), and after grouping, 5 groups per cage Up to group animals were raised. Solid feed (MF, manufactured by Oriental Yeast Co., Ltd.) was used as the feed, and tap water was freely consumed as the drinking water.
- MF manufactured by Oriental Yeast Co., Ltd.
- the grouping was performed on the day of administration of the test solution so that the average body weight of each group was almost uniform by random sampling.
- the group composition there were four groups: a sham operation group, a vehicle control group, an extract A and an extract W administration group.
- the dose was calculated as 10 mL / kg based on the body weight of the administration day so that the dose of extract A was 500 mg / kg per mouse.
- the extract W administration group the dose was calculated as 10 mL / kg based on the body weight on the administration day so that the dose of extract W was 500 mg / kg per mouse.
- the sham surgery group and the vehicle control group were administered with 10 mL / kg of 0.5% (w / v) methylcellulose solution.
- test schedule The administration start date of the test solution was taken as the first day of administration, the test solution was administered once a day, and the amyloid ⁇ solution was injected into the mice on the eighth day of administration. Thereafter, a Y-shaped maze test was performed on the 14th day after administration. Each procedure will be described later.
- the administration route was oral administration.
- a test solution is prepared using a disposable syringe syringe (manufactured by Terumo Corporation) equipped with a disposable oral sonde for mice (manufactured by Fuchigami Instrument Co., Ltd.). Orally administered.
- the test solution was mixed by inversion every time one animal was administered and then aspirated into a syringe.
- the test solution was administered after the amyloid ⁇ solution injection on the amyloid ⁇ solution injection day, and the test solution was administered 30 minutes before the measurement on the Y-shaped maze test day.
- the mouse was anesthetized by intraperitoneally administering pentobarbital sodium (manufactured by Tokyo Chemical Industry Co., Ltd.) to the mouse at 40 mg / kg (dose volume: 10 mL / kg).
- pentobarbital sodium manufactured by Tokyo Chemical Industry Co., Ltd.
- pentobarbital sodium manufactured by Tokyo Chemical Industry Co., Ltd.
- levobupivacaine hydrochloride Bobscaine (registered trademark) 0.25% injection, manufactured by Maruishi Pharmaceutical Co., Ltd.
- the scalp was incised to expose the skull, and a dental drill was used to drill a stainless steel pipe insertion hole in the skull 1 mm (right side) laterally and 0.2 mm behind the bregma.
- a stainless steel pipe connected to a silicon tube having a diameter of 0.5 mm and a microsyringe was inserted vertically to a depth of 2.5 mm from the bone surface.
- 3 ⁇ L (6 nmol / 3 ⁇ L) of amyloid ⁇ solution was injected into the ventricle over 3 minutes with a microsyringe pump.
- 3 ⁇ L of water for injection was injected into the sham operation group by the same method.
- the stainless steel pipe was inserted and left still for 3 minutes, and the stainless steel pipe was slowly removed. Thereafter, the skull hole was closed with a non-absorbable bone marrow hemostatic (Nestop (registered trademark), manufactured by Alfresa Pharma Co., Ltd.), and the scalp was sutured.
- Non-Patent Document 1 A Y-shaped maze test (for example, Non-Patent Document 1), which is an evaluation method for learning and memory behavior, and particularly known as an evaluation method for short-term memory, was performed.
- the length of one arm is 39.5cm
- the width of the floor is 4.5cm
- the height of the wall is 12cm
- a maze manufactured by Unicom
- the illuminance on the floor of the apparatus was adjusted to 10 to 40 lux. Evaluation was carried out 30 minutes after administration of the test solution.
- a Y-shaped maze test was performed on each group of mice, and the total number of entries, the number of spontaneous alternation actions, the average value of the voluntary alternation action rate, and the standard error were calculated.
- the significant difference test compared between two groups of a sham operation group and a vehicle control group, and a vehicle control group and an extract administration group.
- F-test was used for equivariance test.
- Student's t test was used, and for unequal variance, Aspin-Welch test was conducted.
- the significance level was set at 1% risk.
- a commercially available statistical program SAS system, SAS Institute Japan Co., Ltd. was used for the significant difference test. The results are shown in Table 1 and FIG.
- the voluntary alternation behavior rate was lower in the vehicle control group than in the sham operation group, and a significant difference was observed (p ⁇ 0.01).
- the voluntary alternation behavior rate of the extract A administration group was high, and a significant difference was observed (p ⁇ 0.01).
- the voluntary alternation behavior rate of the extract W administration group was higher than that of the vehicle control group, and a significant difference was observed (p ⁇ 0.05).
- Example c1 Deodorizing effect on isovaleric acid> After 0.17 g of Extract A was dissolved in 1 mL of 20% ethanol, 1 mL of water was added and further dissolved. 9 mL of water was added to 1 mL of the solution after dissolution to prepare a solution containing Extract A (Dissolution (A1)). A 15 mL centrifuge tube was charged with 10 mL of 1 ppm (w / v) aqueous solution of isovaleric acid and 0.2 mL of the solution (A1) to prepare sample solution (1) (final concentration of extract A: 0) .017% (w / v)). As a control, a mixed solution of 10 mL of an aqueous solution of 1 ppm (w / v) isovaleric acid and 0.2 mL of water was prepared (control (1)).
- Example c2 Deodorizing effect on acetic acid> A 15 mL centrifuge tube was charged with 10 mL of 0.1% acetic acid and 0.2 mL of the above-mentioned solution (A1) to prepare a sample solution (2) (final concentration of extract A: 0.017% (w / V)). As a control, a mixed solution of 10 mL of 0.1% acetic acid and 0.2 mL of water was prepared (Control (2)). For each of the sample liquid (2) and the control (2), the odor level was evaluated by a panel of nine persons based on the same evaluation criteria as in Test Example c1.
- Example c3 Deodorizing effect on methyl mercaptan>
- a 15 mL centrifuge tube was charged with 10 mL of an aqueous solution of 0.1 ppm methyl mercaptan and 0.2 mL of the above-described solution (A1) to prepare a sample solution (3) (final concentration of extract A 0.017 % (W / v)).
- a sample solution (3) final concentration of extract A 0.017 % (W / v)
- a mixed solution of 10 ppm of 0.1 ppm (w / v) methyl mercaptan and 0.2 mL of water was prepared (Control (3)).
- the odor level was evaluated by a panel of nine persons based on the same evaluation criteria as in Test Example c1.
- Example c4 Deodorizing effect on trimethylamine>
- a 15 mL centrifuge tube was charged with 10 mL of an aqueous solution of 1 ppm (w / v) trimethylamine and 0.2 mL of the above-described solution (A1) to prepare a sample solution (4) (final concentration of extract A: 0) .017% (w / v)).
- a sample solution (4) final concentration of extract A: 0) .017% (w / v)
- control a mixed solution of 10 mL of an aqueous solution of 1 ppm (w / v) trimethylamine and 0.2 mL of water was prepared (control (4)).
- the odor level was evaluated by a panel of 10 persons based on the same evaluation criteria as in Test Example c1.
- Example c5 Deodorizing effect on diacetyl> A 15 mL centrifuge tube was charged with 10 mL of an aqueous solution of 2 ppm (w / v) diacetyl and 0.2 mL of the above-mentioned solution (A1) to prepare a sample solution (5) (final concentration of extract A: 0) .017% (w / v)). As a control, a mixed solution of 10 mL of an aqueous solution of 2 ppm (w / v) diacetyl and 0.2 mL of water was prepared (control (5)). For each of the sample liquid (5) and the control (5), the degree of odor was evaluated by a panel of nine persons based on the same evaluation criteria as in Test Example c1.
- Example c6 Deodorizing effect on Nonenal>
- 10 mL of 0.01 ppm (w / v) nonenal aqueous solution and 0.4 mL of the above-mentioned solution (A1) were added to prepare a sample solution (6) (end of extract A). Concentration 0.034% (w / v)).
- a mixed solution of 10 mL of 0.01 ppm (w / v) nonenal aqueous solution and 0.4 mL of water was prepared (control (6)).
- the odor level was evaluated by a panel of 11 persons based on the same evaluation criteria as in Test Example c1.
- Example c7 Deodorizing effect on ammonium thioglycolate>
- 10 mL of 0.85% (w / v) ammonium thioglycolate aqueous solution and 0.4 mL of the above-mentioned solution (A1) were added to prepare a sample solution (7) (extraction).
- control (7) a mixed solution of 10 mL of an aqueous solution of 0.85% (w / v) ammonium thioglycolate and 0.4 mL of water was prepared (control (7)).
- the odor level was evaluated by 10 panels based on the same evaluation criteria as in Test Example c1.
- Example c8 Deodorizing effect on monoethanolamine thioglycolate>
- 10 mL of an aqueous solution of 0.085% (w / v) thioglycolic acid monoethanolamine and 0.4 mL of the above-mentioned solution (A1) were added to prepare a sample solution (8).
- the final concentration of Extract A is 0.034% (w / v)).
- a mixture of 10 mL of an aqueous solution of 0.085% (w / v) thioglycolic acid monoethanolamine and 0.4 mL of water was prepared (control (8)).
- the odor level was evaluated by a panel of 10 persons based on the same evaluation criteria as in Test Example c1.
- ⁇ Test Example c9 Deodorizing effect on tobacco odor> After 0.17 g of Extract A was dissolved in 1 mL of 20% ethanol, 1 mL of water was added and further dissolved. 30 mL of water was added to 0.3 mL of the solution after dissolution to prepare a solution containing Extract A (Dissolution (A2)). The 5 L Erlenmeyer flask was turned upside down, and a lit cigarette was placed in the mouth of the Erlenmeyer flask about 5 cm, and tobacco smoke was collected for about 30 to 40 seconds. Three pieces of 10 cm ⁇ 10 cm cloth (cotton towels) were put in an Erlenmeyer flask in which smoke was collected and sealed quickly, and the cloth was made to smoke while shaking the flask.
- test fabric was sprayed with water 5 times (about 0.75 ml) (control (9)).
- control (9) a test fabric was sprayed with water 5 times (about 0.75 ml) (control (9)).
- the odor level was evaluated by a panel of nine persons based on the same evaluation criteria as in Test Example c1.
- ⁇ Test d Test as an anti-aging agent> ⁇ Test Example d1: MMP-1 production inhibition test> The MMP-1 production inhibitory effect of Extract A and Extract W was examined by evaluating the inhibitory action of MMP-1 on normal human fibroblasts.
- Test Example d1-1 As a medium for culturing normal human fibroblasts, Dulbecco's modified MEM medium containing 5% calf serum (manufactured by Kurashiki Boseki Co., Ltd., hereinafter also referred to as “5% FBS-DMEM medium”) was used. A medium containing Extract A at a concentration shown in Table 2 in 5% FBS-DMEM medium was prepared, and this was used as a test sample-containing medium.
- 5% FBS-DMEM medium Dulbecco's modified MEM medium containing 5% calf serum
- Normal human fibroblasts (Kurashikibo Co., Ltd.) were seeded in a 96-well microplate together with 5% FBS-DMEM medium at a density of 2.0 ⁇ 10 4 cells / well. 24 hours after seeding, the 5% FBS-DMEM medium in the microplate was replaced with a test sample-containing medium containing Extract A. After culturing for 24 hours after the medium exchange, the test sample-containing medium was replaced with Hanks buffer (containing Ca 2+ and Mg 2+ , HBS (+)). Immediately thereafter, the medium was replaced with a fresh test sample-containing medium, and further cultured for 24 hours. After culturing, the culture supernatant was collected and subjected to ELISA.
- Hanks buffer containing Ca 2+ and Mg 2+ , HBS (+)
- ELISA was performed by the sandwich method, and was performed by the following method.
- Anti-human MMP-1 antibody was added to a high-adsorption ELISA plate, coated overnight at room temperature, and then blocked with 1% bovine albumin (BSA) for 1 hour. After blocking, culture supernatant and calibration curve MMP-1 were added and incubated at room temperature for 2 hours, then Anti-human MMP-1 biotinylated antibody was added and incubated at room temperature for 1.5 hours. Further, streptavidin HRP was added and incubated at room temperature for 30 minutes.
- BSA bovine albumin
- the amount of MMP-1 in the culture supernatant was calculated from a calibration curve prepared with commercially available MMP-1.
- the cultured cells were dissolved in 0.5% (V / V) Triton X-100 buffer, and the total protein amount was quantified by the BCA method.
- the amount of MMP-1 produced per unit protein was calculated by dividing the amount of MMP-1 in the culture supernatant by the total amount of protein in the cells. The results are shown in Table 2.
- Test Example d1-2 In Test Example d1-1, the test sample-containing medium was replaced with Hanks buffer (Ca 2+ , Mg 2+ contained, HBS (+)), and then irradiated with ultraviolet A wave (UVA) at a dose of 4 J / cm 2. Thus, the amount of MMP-1 produced per unit protein was calculated in the same manner as in Test Example d1-1 except that the medium was immediately replaced with a fresh test sample-containing medium. The results are shown in Table 2.
- Test Example d1-3 In Test Example d1-2, except that Extract A was changed to Extract W and the dose of ultraviolet A wave was changed to 5 J / cm 2 , the same method as in Test Example d1-2 was performed per unit protein amount. MMP-1 production was calculated. The results are shown in Table 2.
- test solution 50 ⁇ L was added to each well of a 96-well microplate. Also, 100 mmol / L Tris-HCl buffer (pH 8.0) containing 5 mmol / L Suc-Ala-Ala-Ala-pNA was prepared and added in 100 ⁇ L aliquots. 50 ⁇ L of the elastase crude enzyme solution was added to each well to obtain a reaction solution. For the reaction solution immediately after addition of the elastase crude enzyme solution, the absorbance at 405 nm was measured as the absorbance before the reaction (blank absorbance).
- Test Example d2-2 Except that the extract A was changed to the extract W, the elastase activity inhibition rate of the extract W was determined by the same method as in Test Example d2-1. In addition, 12.25 mmol / L EDTA was used as a positive control. Also in the extract W, the elastase activity inhibitory effect was recognized.
- Extract A was dissolved in water to prepare a medium containing Extract A in 1% FBS-DMEM medium at the concentrations shown in Table 4, and this was used as a test sample-containing medium.
- As a positive control 5% FBS-DMEM medium was used.
- Normal human fibroblasts (Kurashikibo Co., Ltd.) were seeded in a 96-well microplate together with 5% FBS-DMEM medium at a density of 2.0 ⁇ 10 4 cells / well. 24 hours after seeding, the 5% FBS-DMEM medium in the microplate was replaced with a test sample-containing medium containing Extract A. After culturing for 48 hours after the medium change, 1% FBS containing 0.4 mg / mL 3- (4,5-dimethylthiazolyl-2-yl) -2,5-diphenyltetrazolium bromide (MTT) -Change to DMEM medium and incubate for 2 hours.
- MTT 4,5-dimethylthiazolyl-2-yl
- the medium was removed, and blue formazan produced by adding 2-propanol was extracted. Absorbance of the extract at 550 nm was measured and used as the amount of blue formazan.
- concentration of extract A based on the total amount of the test sample-containing medium
- concentration of extract A 0 ⁇ g / mL (control)
- the ratio (percentage) of the amount of blue formazan when using extract A to the amount of blue formazan produced is calculated.
- the cell activation rate was used. It can be said that the larger the value of the cell activation rate, the higher the cell activation effect. The results are shown in Table 4.
- Extract A which is a decomposition extract of bagasse, was dissolved in distilled water to a concentration of 100 mg / mL to prepare a test solution stock solution.
- This test stock solution was diluted with distilled water to prepare a 0.01-100 mg / mL solution. This was used as a test sample.
- an aqueous solution (concentration: 3.0 mg / mL) of aminoguanidine which is a saccharification reaction inhibitor was prepared.
- the AGE generation inhibition rate (hereinafter also simply referred to as “inhibition rate”) is a reaction solution in which the fluorescence intensity of the reaction solution to which the sample is added in the saccharification reaction is F 1 and distilled water is added instead of the glucose aqueous solution and incubated.
- Fluorescence AGEs inhibition rate (%) (1 ⁇ (F 1 ⁇ F 2 ) / (F 3 ⁇ F 4 )) ⁇ 100
- FIG. 6 shows the inhibition rate of fluorescent AGEs (HSA) at the final reaction concentration (0.1 mg / mL, 0.3 mg / mL or 1 mg / mL) of the decomposition extract of bagasse (Extract A).
- the extract A increased in inhibition rate in a concentration-dependent manner and exhibited anti-glycation activity (fluorescence AGEs (HSA) production inhibitory action).
- the inhibition rate of fluorescent AGEs (HSA) at 0.3 mg / mL of aminoguanidine as a positive control was 81.2 ⁇ 1.4%. It was confirmed that aminoguanidine has anti-glycation activity (fluorescence AGEs (HSA) production inhibitory action).
- IC 50 (50% production inhibitory concentration) calculated from the inhibition rate of the sample at each concentration of Extract A was 0.19 mg / mL, and in the human serum albumin model, Extract A has anti-glycation activity. It was shown that.
- Extract A a decomposition extract of bagasse, was dissolved in distilled water to 20 mg / mL to prepare a sample for testing.
- the amount of benzoic acid in the reaction solution was determined by subtracting the amount of benzoic acid in the sample measured separately. Since 1 mol of PPD produces 1 mol of benzoic acid, the crosslinking cleavage rate was calculated by the following formula.
- Crosslink cutting rate (%) ⁇ (AB) / C ⁇ ⁇ 100
- Extract W which is a decomposition extract of bagasse
- 50% DMSO 50% DMSO
- This solution was serially diluted with 50% DMSO to prepare a test sample.
- a 10 mmol / L PTB N-phenacylthiazolbromide
- the crosslinking cleavage rate was calculated by the following formula.
- the relative value of the crosslink cutting is the value (%) of the crosslink cutting rate at each concentration when the crosslink cutting rate of PTB is 100.
- Shimadzu ultra-high performance liquid chromatograph Nexera system made by Shimadzu Corporation was used as a measuring apparatus.
- Crosslink cutting rate (%) ⁇ (AB) / C ⁇ ⁇ 100
- C Amount of PPD subjected to reaction (base mass)
- Extract A was dissolved in water to prepare a 50 mg / mL test solution stock solution. This test solution stock solution was diluted with a buffer solution shown in Table 5 below to prepare test solutions having specimen concentrations of 2000, 1000, and 500 ⁇ g / mL, respectively.
- DNP labeled human serum albumin was added and it was made to react at 37 degreeC for further 3 hours.
- what added only buffer solution without adding a test solution is what added untreated control (comparative example f1) and wortmannin (Wako Pure Chemical Industries, Ltd.) so that it might become final concentration of 25 nmol / L.
- a similar test was performed as a positive control.
- a buffer solution and DNP-labeled human serum albumin were sequentially added and reacted in the same manner as an antigen unstimulated control.
- a cell lysis buffer (Lysis buffer) was added to the cells and allowed to stand at room temperature for 10 minutes to obtain a cell lysate.
- P-Nitrophenyl-2-acetamido-2-deoxy- ⁇ -D-glucoplanoside solution (hereinafter referred to as substrate solution) was added to the cell supernatant and cell lysate, respectively, and reacted at 37 ° C. for 25 minutes. Thereafter, glycine buffer was added to stop the reaction.
- a sample blank was prepared by adding a glycine buffer to the cell supernatant and the cell lysate, reacting at 37 ° C. for 25 minutes, and adding a substrate solution.
- release rate absorbance on the cell supernatant side / (absorbance on the cell supernatant side + absorbance on the cell solution side)
- Degranulation rate (%) ⁇ average value of (release rate of test solution ⁇ release rate of antigen-unstimulated control) / (release rate of untreated control ⁇ release rate of antigen-unstimulated control) ⁇ ⁇ 100
- Example f3 was 77 ⁇ 11.1.
- Extract W was dissolved in ethanol to prepare a 50 mg / mL test solution stock solution. This test solution stock solution was diluted with the buffer solution shown in Table 5 above to prepare test solutions having specimen concentrations of 1000, 500, and 250 ⁇ g / mL, respectively.
- Example f4 [RBL-2H3 cell degranulation inhibition test] Using the prepared test solution containing the extract W, the final concentrations of the test solution were 500 ⁇ g / mL (Example f4), 250 ⁇ g / mL (Example f5), and 125 ⁇ g / mL during the test operation in Test Example f1 described above.
- the degranulation rate was calculated by the same method as in Test Example 1 except that the test solution was added so as to be Example f6).
- Comparative example f1 is an untreated control similar to test example f1.
- Example f6 The results of calculating the degranulation rate are shown in FIG.
- the degranulation rate was 100 ⁇ 2.9 for Comparative Example f1, whereas 12 ⁇ 1.6 for Example f4, 47 ⁇ 4.9 for Example f5, and 80 ⁇ 6.
- ⁇ Test g Test as antihypertensive agent> ⁇ Angiotensin converting enzyme inhibition test> Angiotensin converting enzyme inhibition test was conducted based on the method of Nakano et al. (Nakano et al, Biosci. Biotechnol. Biochem., 70, 1118-1126 (2006)). Extract A and Extract W, 1.0 g each, were extracted with 20 ml of 50% (V / V) ethanol solution and then diluted appropriately with 0.1 mol / L Hepes buffer (pH 8.3). Test solutions of the indicated concentrations were prepared.
- IC 50 of Extract A is 0.95 mg / mL based on the total amount of the test solution (final concentration based on the total amount of the specimen for measurement: 0.16 mg / mL), and IC 50 of Extract W is 0.52 mg / mL based on the total amount of the test solution. mL (final concentration based on the total amount of the sample for measurement: 0.087 mg / mL).
- Extract A> [Soy milk]
- soy milk (trade name: amazing soybean non-adjustable type, manufactured by Otsuka Chilled Foods Co., Ltd.), extract A solution (solid content concentration 0.3%) is 0 so that the concentration of extract A is as shown in Table 7. .6 g (Example h1-1) or 0.3 g (Example h1-2) was added to prepare test articles (Examples h1-1 and h1-2).
- test product (Example h1-3) was prepared by adding 1.7 g of a solution of extract A (solid content concentration 0.3%) to a vinegar-containing beverage of 50 g of rice vinegar, 200 g of water and 15 g of granulated sugar. .
- Example h2 Flavor Improvement Test Using Extract W> [Soy milk, vinegar-containing beverages, processed beef (meat dumplings)] Test samples (Examples h2-1 to h2-3) were prepared in the same manner as in Test Example h1, except that Extract A was changed to Extract W in Test Example h1.
- An immersion liquid was prepared by adding 4.175 g of a solution of extract W (solid content concentration: 0.3%) to 500 g of 8% saline. After thawing the frozen fillet mackerel, the skin was cut, and then immersed in the immersion liquid for 10 minutes. The both sides of the mackerel were baked for 7 minutes each time using a grill to prepare a test article (Example h2-4). The test article was subjected to sensory evaluation after it was frozen once in a microwave oven.
- [Lactic acid bacteria beverage] Commercial beverage (trade name: Mill Mill, manufactured by Yakult Co., Ltd.) 100 parts by weight, 10% acetic acid 0.275 parts by weight, aspartame (Ajinomoto Co., Ltd.) 0.007 parts by weight, and 1% solution of ⁇ -carotene (Mitsubishi Chemical)
- a lactic acid bacteria beverage containing 0.015 parts by mass of Foods Co., Ltd. was prepared.
- 0.075 g of the extract W solution solid concentration 0.3%) was added to 100 g of this lactic acid bacteria beverage to prepare a test article (Example h2-5).
- ⁇ Test Example h3 Comparative test with sweet potato-derived extract> [Manufacture of sweet potato-derived extract] 7500L of sweet potato soup (purified soup obtained from lime purification in raw sugar manufacturing factory, Okinawa, solid content 14%) 7500L is used as a cartridge filter (Advantech Co., Ltd., Cotton Wind Cartridge Filter, TCW) -10-CSD type) to obtain a filtered product of purified juice.
- a cartridge filter Advancedtech Co., Ltd., Cotton Wind Cartridge Filter, TCW) -10-CSD type
- Examples h3-5 to h3-6 and Examples h3-9 to h3-10 (meat dumplings and lactic acid bacteria beverages), a diluted solution of sweet potato extract was used as the extract A in the preparation method of the control product described in Table 16.
- the test article was prepared by changing the solution to the above solution or the extract W solution (solid content concentration: 0.3%).
- Example h4 Evaluation by taste recognition device> The above-mentioned Extract A and sweet potato extract were added to a 0.2% acetic acid aqueous solution, and a 0.2% acetic acid aqueous solution having a final solid content concentration of 45 ppm, 90 ppm, and 180 ppm (Example h4-1) To h4-3 and Comparative Examples h4-1 to h4-3) were prepared. The acidity of this acetic acid solution was measured using a taste recognition device (TS-5000Z, manufactured by Intelligent Sensor Technology). When measuring with the said taste recognition apparatus, it measured with the taste (relative value) of the acidity sensor (CAO).
- TS-5000Z manufactured by Intelligent Sensor Technology
- Table 22 shows the relative value of the acidity of each aqueous acetic acid solution with the acidity of the aqueous acetic acid solution (control) to which the extract A and sweet potato extract were not added as 0.
- a relative value of the sourness of 0 or less indicates that the taste (acidity) felt by acetic acid is reduced as compared with the 0.2% acetic acid aqueous solution as a control.
- Growth medium composition DMEM medium, 10% FBS, antibiotic added
- Differentiation medium composition DMEM medium, 0.5% FBS, antibiotic added
- C2C12 cells were asleep in a T-75 flask (75 cm 2 U-shaped Cantoneck cell culture flask, Corning) using a growth medium.
- the T-75 flask was placed in a CO 2 incubator (5% CO 2 , 37 ° C., wet) to culture C2C12 cells.
- the medium was changed every other day, and when the cells reached 80% confluence, the cells were collected and used for the test.
- a plate coated with gelatin by the coating method shown below was used as a well plate for culturing C2C12 cells.
- a 0.75% gelatin aqueous solution was sterilized by an autoclave.
- differentiation medium containing extract A (Examples i1-1 to i1-3), differentiation medium containing extract W and 1% (v / v) ethanol (Examples i1-4 to i1) -6), differentiation medium containing no extract (Comparative Example i1-1), differentiation medium containing no extract and containing 1% (v / v) ethanol (Comparative Example i1-2), or less at a concentration of 50 ⁇ M
- the culture medium was changed to a differentiation medium (positive control i1-1) containing veratrol and 1% (v / v) ethanol, and cultured. The culture was terminated 48 hours after the start of the culture.
- Examples i1-1 to i1-6 were used as test samples for Examples i1-1 to i1-6, Comparative Examples i1-1 to i1-2, or positive control i1-1.
- the final concentration of the bagasse degradation extract in the differentiation medium is shown in Table 23.
- a differentiation medium containing 5 ⁇ g / mL Hoechst (nuclear staining reagent) or 10 ⁇ g / mL rhodamine (mitochondrial activity staining reagent) was added to the well plate and incubated at 37 ° C. for 30 minutes. . Thereafter, the fluorescence intensity was measured with a fluorescence plate reader. Mitochondrial activity per cell was calculated by dividing the fluorescence intensity of the differentiation medium containing rhodamine by the fluorescence intensity of the differentiation medium containing Hoechst.
- test samples of Examples i1-1 to i1-6 to which an extract was added as compared with the test sample of Comparative Example i1-1 or i1-2 to which no bagasse decomposition extract was added
- mitochondrial activity per cell increased. Since the mitochondrial activity also increased in the positive control i1-1, it can be said that the test was conducted without any problem.
- mitochondrial activity was also evaluated by the same method as described above for the test samples after 72 hours of culture.
- mitochondrial activity in the test sample of Comparative Example i1-2 after culture for 72 hours was 100% (100.0 ⁇ 1.6%)
- the mitochondrial activity in the test sample of Example i1-5 was 101.7 ⁇ 1.
- the mitochondrial activity in the test sample of Example i1-6 was 115.0 ⁇ 2.4%.
- the test samples of Example i1-5 and Example i1-6 to which the extract was added were cultured for 72 hours. Also increased mitochondrial activity per cell.
- Test Example i1-2 Mitochondrial activation test (evaluation of the amount of mitochondria per cell)] Myotubes were cultured by the same method as in Test Example i1-1. These were used as test samples for Examples i1-7 to i1-11, Comparative Examples i1-3 to i1-4, or Positive Control i1-2. In Examples i1-7 to i1-11, the final concentration of the bagasse degradation extract in the differentiation medium is shown in Table 24.
- a differentiation medium containing 5 ⁇ g / mL Hoechst (nuclear staining reagent) and 500 nM mitotracker (mitochondrial staining reagent) was added to the well plate, and the same method as in Test Example i1-1 was used to measure the fluorescence intensity.
- the amount of mitochondria per cell was calculated by dividing the fluorescence intensity of the differentiation medium containing mitotracker by the fluorescence intensity of the differentiation medium containing Hoechst.
- the relative values (%) in the test samples of Examples i1-7 to i1-8 when the amount of mitochondria in the test sample of Comparative Example i1-3 was 100% were determined.
- Example i1-8 and Example i1-11 the amount of mitochondria was also evaluated for the test samples after 72 hours of culture by the same method as described above.
- the amount of mitochondria in the test sample of Comparative Example i1-3 after 72 hours of culture was 100% (100.0 ⁇ 3.4%)
- the amount of mitochondria in the test sample of Example i1-8 was 104.9 ⁇ 4. .5%.
- the amount of mitochondria in the test sample of Comparative Example i1-4 was 100% (100.0 ⁇ 0.8%)
- the amount of mitochondria in the test sample of Example i1-11 was 107.9 ⁇ 1.9%. Met.
- differentiation medium containing extract A (Examples i2-1 to i2-2), differentiation medium containing no extract (Comparative Example i2-1), 0.1% (v / v) containing no extract
- Differentiation medium containing DMSO (Comparative Example i2-2), or 0.5 ⁇ M LDN-193189 (4- (6- (4- (piperazin-1-yl) phenyl) pyrazolo [1,5-a] pyrimidine-3- Yl) quinoline) and 0.1% (v / v) DMSO were replaced with differentiation medium (positive control i2).
- the cells were cultured for 3 days, and after fixation, immunostaining was performed with an anti-myosin heavy chain (MHC) antibody.
- MHC anti-myosin heavy chain
- 4% -paraformaldehyde / phosphate buffer solution was added to the cells after completion of the culture at 100 ⁇ L / well, and the cells were allowed to stand at 4 ° C. for 15 minutes. After standing, the plate was washed 3 times with Dulbecco's PBS (DPBS), and then subjected to blocking treatment with DPBS containing 0.1% Triton-X and 3% bovine serum albumin (BSA) for 1 hour at room temperature. Subsequently, a 3% BSA / DBPS mixed solution containing a primary antibody (anti-myosin heavy chain antibody: diluted 300-fold) was added and reacted at 4 ° C. overnight.
- DPBS Dulbecco's PBS
- BSA bovine serum albumin
- Example i2-1 to i2-2 the test samples of Examples i2-1 to i2-2 to which the bagasse decomposition extract was added were compared with the test sample of Comparative Example i2-1 to which no bagasse decomposition extract was added. Fusion index increased.
- Comparative Example i2-1 two-sided test using Student's T test
- the test sample of Example i2-2 was significantly different from the test sample of Comparative Example i2-1. Increased (p ⁇ 0.01).
- the Fusion index increased compared to the sample of Comparative Example i2-2, it can be said that the test was performed without any problem.
- (cell) Mouse calvaria-derived cells MC3T3-E1 (RIKEN Cell Bank, RCB1126)
- Alkaline phosphatase activity measurement kit (LabAssay ALP, product number 291-58601, Wako Pure Chemical Industries, Ltd.) Protein mass measurement kit (Micro BCA Protein Assay Reagent Kit, product number 23235, PIERCE) Cell Lysis / Protein Extraction Reagent (Cell-LyEX1, Product No. 300-34761, Wako Pure Chemical Industries, Ltd.) 10% neutral buffered formalin solution (Product No. 062-1661, Wako Pure Chemical Industries, Ltd.) Calcein AM (product number PK-CA707-80011, PromoKine) Recombinant bone morphogenetic protein (Bone Morphogenetic Protein-2 (BMP-2), R & D Systems)
- MC3T3-E1 cells were asleep in a T-75 flask (75 cm 2 U-shaped Cantoneck cell culture flask, Corning) using growth medium.
- the T-75 flask was placed in a CO 2 incubator (5% CO 2 , 37 ° C., wet) to culture C2C12 cells.
- the medium was changed every other day, and when the cells reached 80% confluence, the cells were collected and used for the test.
- osteoblast differentiation promotion test The promotion of osteoblast differentiation in MC3T3-E1 cells was confirmed using alkaline phosphatase (ALP) activity, which is one of osteoblast differentiation markers, as an indicator.
- ALP alkaline phosphatase
- Pre-cultured MC3T3-E1 cells were adjusted with medium to 1.2 ⁇ 10 5 cells / 0.2 mL / well and seeded in a 48-well plate.
- Example j1-1 medium containing 100 ⁇ g / mL extract A
- Example j1-2 medium containing 100 ⁇ g / mL extract W and 1% (v / v) ethanol
- Example j1-2 including extract No medium (Comparative Example j1-1), no extract, medium containing 1% (v / v) ethanol (Comparative Example j1-2), or medium containing BMP-2 (positive control j1)
- the culture was replaced and cultured for 7 days, 14 days and 21 days, respectively. After each day of culturing, the cells were washed once with PBS and stored in a frozen state along with the plate. The medium was changed every 3-4 days.
- the cells after cryopreservation were washed with PBS, and then lysed with 100 ⁇ L / well of a cell lysing agent (Cell-LyEX1 containing 2 mM phenylmethylsulfonyl fluoride (PMSF)).
- the plate was stirred at room temperature for 30 minutes and then centrifuged, and a solution obtained by diluting the supernatant 5 times was used as a measurement sample.
- the amount of ALP in the cells was measured using an alkaline phosphatase activity measurement kit (LabAssay ALP). In this kit, ALP activity is measured from the amount of p-nitrophenol per unit protein produced within a certain time.
- Example j1-1 and Example j1- containing bagasse decomposition extract were used. ALP activity was increased in 2 samples. It can be said that the higher the ALP activity, the more the osteoblast differentiation is promoted. Further, when a significant difference test with Comparative Example j1-2 (two-sided test using Student's T test) was performed, the samples of Example j1-1 and Example j1-2 were found on the 14th or 21st day of culture. As compared with the test sample of Comparative Example j1-2, ALP activity was significantly increased. Since the ALP activity also increased in the positive control j1, it can be said that the test was conducted without any problem.
- Test Example j2 Osteoclast differentiation inhibition test 1> [material] In Test Example j2, the following materials were used. (cell) Human osteoclast progenitor cells (Cosmo Bio Corporation PT-267 Lot. RBW-F-OSH-HBV)
- FIG. 11A shows the results of Example j2-1
- FIG. 11B shows the results of Comparative Example j2-1
- FIG. 11C shows the results of the positive control j2.
- Example j2-1 As shown in FIG. 11, in Example j2-1, a decrease in multinucleated mature osteoclasts was observed. On the other hand, in Comparative Example j2-1 that did not contain the bagasse degradation extract, no decrease in osteoclasts was observed. Since the decrease in osteoclasts was also observed in the positive control j2, it can be said that the test was conducted without any problem.
- Example j3-1 a medium containing Extract A at a concentration shown in Table 27 (Example j3-1), a medium containing no extract (Comparative Example j3-1), or a positive target Human osteoclast precursor cells were cultured using media containing melatonin at the concentrations shown in Table 27 (positive controls j3-1 to j3-2).
- the percentage (%) of mononuclear cells was calculated by dividing the number of mononuclear cells in the microscopic field by the total number of cells in the field. The results are shown in Table 27.
- Example j3-1 containing the bagasse decomposition extract As shown in Table 27, the proportion of mononuclear in Example j3-1 containing the bagasse decomposition extract was increased as compared with Comparative Example j3-1 not containing the bagasse decomposition extract. It can be said that the higher the proportion of mononuclear cells, the more osteoclast differentiation is suppressed. In the positive controls j3-1 to j3-2, the ratio of mononuclear cells also increased, so it can be said that the test was conducted without any problem.
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Abstract
Description
(1)花粉やダニなどの抗原(アレルゲン)が生体内に侵入すると、ヘルパーT細胞(Th2細胞)が指令を出しB細胞を免疫グロブリンE(IgE)抗体産生細胞へと分化させる。
(2)IgE抗体産生細胞からその抗原に特異的なIgE抗体が産生される。
(3)IgE抗体が肥満細胞又は好塩基球に結合し、そこに再び抗原が結合することによりヒスタミン、ロイコトリエン等の化学伝達物質が分泌(脱顆粒)され、アレルギー症状が発現する。
「バガス」とは、典型的には原料糖製造工程における製糖過程で排出されるバガスをいう。原料糖工場における製糖過程で排出されるバガスには、最終圧搾機を出た最終バガスだけではなく、第1圧搾機を含む以降の圧搾機に食い込まれた細裂甘蔗をも含む。好適なバガスは、原料糖工場において圧搾工程により糖汁を圧搾した後に排出されるバガスである。当該バガスは、甘蔗の種類、収穫時期等により、その含まれる水分、糖分及びそれらの組成比が異なるが、本発明においては、これらのバガスを任意に用いることができる。さらに、一実施形態では、原料のバガスとして、原料糖工場と同様に、例えば黒糖製造工場において排出される甘蔗圧搾後に残るバガス、又は実験室レベルの小規模な実施により甘蔗から糖液を圧搾した後のバガスも用いることができる。
本明細書における肥満抑制剤は、肥満を抑制する作用を備える組成物である。肥満を抑制する作用とは、例えば、脂肪細胞における脂肪の蓄積を抑制する作用(脂肪蓄積抑制作用)、脂肪細胞に蓄積した脂肪の分解を促進する作用(脂肪分解促進作用)、又は脂肪細胞の増殖を抑制する作用(脂肪細胞増殖抑制作用)等であってよい。すなわち、本明細書における肥満抑制剤は、脂肪蓄積抑制剤、脂肪分解促進剤、又は脂肪細胞増殖抑制剤等であってよい。
本発明の抗認知症剤は、抗認知症作用を有するものである。本発明における「抗認知症作用」とは、認知症の発症を未然に防止する作用、認知症の発症を遅延させる作用、一度発症した認知症を発症時の状態から回復させる作用を含む概念である。本発明の抗認知症剤が対象とする認知症は、アルツハイマー型認知症であってもよい。アルツハイマー型認知症は、前脳基底部のマイネルト核におけるアセチルコリン作動性神経細胞の脱落により引き起こされる病態(コリン仮説)と、アミロイドβタンパク質の蓄積によって引き起こされる病態(アミロイド仮説)とがある。両者はそれぞれ別々の病態を示すものであり、同一の病態を別の角度から見たものではない。本発明の抗認知症剤が対象とする認知症はいずれの説に基づくアルツハイマー型認知症であってもよく、好ましくは、アミロイド仮説に基づくアルツハイマー型認知症である。すなわち、本発明の抗認知症剤が対象とする認知症は、アミロイドβタンパク質の蓄積に起因するアルツハイマー型認知症であってよい。言い換えると、本発明は、アルツハイマー型認知症の抗認知症剤、アミロイド仮説に基づくアルツハイマー型認知症の抗認知症剤、又はアミロイドβタンパク質の蓄積に起因するアルツハイマー型認知症の抗認知症剤を提供するということもできる。
本明細書における「消臭剤」は、消臭効果を有する成分(有効成分)を含有するものである。消臭剤は、消臭対象物の臭いを消去又は抑制することにより、消臭効果を発揮するものであってもよく、消臭対象物の臭いをマスキングすることで、消臭効果を発揮するもの(マスキング剤)であってもよい。
本発明の抗老化剤は、抗老化作用を有する。抗老化作用は、皮膚の老化を抑制する作用であってよく、より具体的には、加齢、紫外線の照射等による皮膚の機能低下を抑制及び/又は改善する作用であってよい。抗老化作用は、皮膚のしわ、たるみ、硬化等を抑制及び/又は改善する作用であってもよい。
本明細書における抗糖化剤は、抗糖化活性を有するものであり、具体的には、糖化最終生成物(AGEs)の、生成抑制作用(糖化反応阻害作用)、蓄積抑制作用又は分解作用を有するものであってよい。言い換えれば、本実施形態の抗糖化剤は、例えば、糖化最終生成物の生成抑制剤(糖化反応阻害剤)、蓄積抑制剤又は分解剤(分解促進剤)であってよい。
本明細書における抗I型アレルギー剤は、I型アレルギー症状を抑制する作用を備える組成物である。I型アレルギー症状を抑制する作用とは、例えば、花粉症、蕁麻疹、アレルギー性鼻炎、気管支喘息等のI型アレルギーによる症状を緩和、治療、又は予防する作用であってよい。また、I型アレルギー症状を抑制する作用とは、I型アレルギー反応のメカニズムにおける、肥満細胞又は好塩基球の脱顆粒を抑制する作用であってよい。すなわち、本明細書における抗I型アレルギー剤は、肥満細胞又は好塩基球の脱顆粒抑制剤であってもよい。
本発明の抗高血圧剤は、抗高血圧作用を有する。抗高血圧作用は、血圧の上昇を抑制する作用であってよい。
本発明の風味向上剤は、飲食品の風味を向上させる作用を有する。風味とは、味及び臭いが各々存在するとき、又は味と臭いとが複合的に存在するときに感じる感覚、並びに、味と喉への刺激とが複合的に存在するときに感じる感覚の全てを包含する。
本発明の筋肉増強剤は、筋肉増強作用を有する。本明細書における筋肉増強作用には、筋芽細胞から筋管細胞への分化を促進する、筋管細胞分化促進作用、及び筋肉中のミトコンドリアを賦活化させる、ミトコンドリア賦活作用を含む。すなわち本発明は、筋管細胞分化促進剤、又はミトコンドリア賦活剤を提供するということができる。
本発明の骨代謝改善剤は、骨代謝の改善作用を有する。骨代謝の改善作用は、骨形成(新たな骨の形成)を促進する作用、及び過度な骨吸収(骨の破壊)を抑制する作用の少なくとも1種であってよい。これにより、骨形成と骨吸収とのバランスを好適に調整することができ、結果として骨の再構築を進行させやすくすることができる。すなわち、本発明は、骨形成促進剤、及び骨吸収抑制剤を提供するということができ、骨形成及び骨吸収のバランス調整剤を提供するということもできる。
[製造例1]
サトウキビの搾りかすであるバガス15kg(含水率50質量%)及び0.5%(w/w)水酸化ナトリウム水溶液100Lを混合し、150℃の条件でアルカリ処理を行った。アルカリ処理後の混合液を固形分と液分に分離して、液分を約100L得た。分画分子量2500のUF膜(SUEZ社、GH8040F30)を用いて限外濾過を行い、濾過液80Lを得た。合成吸着剤(三菱ケミカル株式会社製、HP-20)1Lを樹脂塔(内径80mm、高さ400mm)に充填し、これに上記の濾過液を、pHを6に調整してから流速10L/時間(SV=10.0(時間-1))で通液した。
サトウキビの搾りかすであるバガス30kg(含水率50質量%)を、200℃、1.8MPaの熱水100Lで水熱処理を行った。前処理後の混合液を固形分と液分とに分離して、液分を約88L得た。分画分子量2500のUF膜(SUEZ社、GH8040F30)を用いて限外濾過を行い、濾過液70Lを得た。合成吸着剤(三菱ケミカル株式会社製、SP-850)1Lを樹脂塔(内径80mm、高さ400mm)に充填し、これに上記の濾過液のうち25Lを、流速20L/時間(SV=20.0(時間-1))で通液した。
<試験例a1:抽出物Aによる肥満抑制試験>
[試験液の調製]
抽出物Aを水に溶解させて、50mg/mLの試験液原液を調製した。この試験液原液をDMEM培地で希釈し、検体濃度2000μg/mLの試験液を調製した。
(細胞の培養)
マウス脂肪前駆細胞である3T3-L1細胞(国立研究開発法人 医薬基盤・健康・栄養研究所)を24ウェルプレートに播種し、新生仔牛血清(10vol%、培地全量基準)及びペニシリン-ストレプトマイシン溶液(1vol%、培地全量基準)を含有するDMEM培地で4日間培養した。培養後、牛胎児血清(10vol%、培地全量基準)及びペニシリン-ストレプトマイシン溶液(1vol%、培地全量基準)を含有するDMEM培地に交換し、Adipogenesis Assay Kit(ケイマンケミカル社)を用いて、3T3-L1細胞を脂肪細胞へと分化させた。この際、調製した2000μg/mLの試験液を、終濃度が1000μg/mL(実施例a1)となるように添加した。3日間培養後、新たに用意した試験液添加培地に交換した。また、Adipogenesis Assay Kitによる分化誘導を行わないものを前駆細胞(未分化a1-1)、試験液を添加しなかったものを未処置対照(比較例a1-1)、塩化ベルベリン(和光純薬株式会社)を終濃度1μg/mLとなるように添加したものを陽性対照(陽性対照a1-1)として同様に試験を行った。これらを更に4日間培養した。
培養後、培養上清を除去し、Adipogenesis Assay Kitの手順に従って細胞を固定した後、脂肪滴を染色するための親油性色素であるオイルレッドO溶液により細胞を染色した。
倒立型位相差顕微鏡を用いて、脂肪細胞に蓄積した脂肪滴を染色した結果を観察した。各サンプルの観察結果を図1に示す。図1において、(a)が前駆細胞(未分化a1-1)、(b)が比較例a1-1、(c)が陽性対照a1-1、(d)が実施例a1の観察結果である。その結果、比較例a1-1の脂肪細胞では、オイルレッドOで染色された脂肪滴の蓄積が観察できたが、実施例a1の脂肪細胞では、比較例a1-1の脂肪細胞と比較して、蓄積している脂肪滴が少なかった。
脂肪滴を観察後、実施例a1、比較例a1-1及び陽性対照a1-1のサンプルに対して、Adipogenesis Assay Kitに含まれる色素抽出液を添加し、脂肪滴に取り込まれたオイルレッドOを抽出した。マイクロプレートリーダー(SpectraMax M2e、モレキュラーデバイス社)を用いて、抽出したオイルレッドOの吸光度を520nmにて測定した。
脂肪蓄積率(%)={(Sa-BLAvr)/(CN-BLAvr)Avr}×100
Sa:各サンプルの吸光度
BLAvr:前駆細胞(未分化a1-1)の吸光度の平均値(n=3)
CN:比較例a1-1(未処置対照)の吸光度
(CN-BLAvr)Avr:CNよりBLAvrを差し引いた値の平均値(n=3)
[試験液の調製]
抽出物Wをエタノールに溶解させて、50mg/mLの試験液原液を調製した。この試験液原液をDMEM培地で希釈し、検体濃度500μg/mLの試験液をそれぞれ調製した。
試験例a1における方法において、調製した抽出物Wを含む試験液を、終濃度が250μg/mL(実施例a2)となるようにそれぞれ添加した以外は、試験例a1と同様の方法によって脂肪蓄積抑制試験を行った。試験例a1と同様に、Adipogenesis Assay Kitによる分化誘導を行わないものを前駆細胞(未分化a1-2)、試験液を添加しなかったものを未処置対照(比較例a1-2)として同様に試験を行った。
試験例a1と同様の方法により、脂肪細胞に蓄積した脂肪滴を染色した結果を観察した。各サンプルの観察結果を図3に示す。図3において、(a)が前駆細胞(未分化a1-2)、(b)が比較例a1-2、(c)が実施例a2の観察結果である。その結果、比較例a1-2の脂肪細胞では、オイルレッドOで染色された脂肪滴の蓄積が観察できたが、実施例a2の脂肪細胞では、比較例a1-2の脂肪細胞と比較して、蓄積している脂肪滴が少なかった。
脂肪滴を観察後、実施例a2及び比較例a1-2について、試験例a1と同様の方法により脂肪蓄積率を算出した。結果を図4に示す。脂肪蓄積率は、比較例a1-2が100±5.6%であったのに対して、実施例a2が20±5.6であった。
[試験溶液の調製]
上記の抽出物A及び抽出物Wについては、試験に供するまで、室温(管理温度:18.0~28.0℃)で粉末の状態で保管した。抽出物を溶解する媒体として注射用水(大塚蒸留水、株式会社大塚製薬工場)を用意した。抽出物の必要量を秤量し、注射用水で溶解した後、所定濃度となるように希釈し、これを試験溶液とした。
アミロイドβ溶液に使用するアミロイドβ(Amyloid-βProtein(25-35)、Polypeptide Laboratories社製)は、試験に供するまで、冷凍(管理温度:-30℃~-20℃(実測値:-27.1℃~-24.0℃)で保管した。アミロイドβを注射用水で2mMとなるように溶解させ、アミロイドβ溶液を調製した。
試験動物として、雄性Slc:ddYマウス(SPF、日本エスエルシー株式会社製)を使用した。当該マウスとして、5週齢のものを入手した。当該マウスは、行動薬理試験に一般的に用いられている動物種で、その系統維持が明らかなものである。入手後1日のマウスの体重範囲は23.8~30.0gであった。入手したマウスについて5日間の予備飼育期間を設けた。
マウスは、管理温度20.0~26.0℃、管理湿度40.0~70.0%、明暗各12時間(照明:午前6時~午後6時)、換気回数12回/時(フィルターを通した新鮮空気)に維持された動物飼育室で飼育した。
予備飼育期間中から群分け日までは、プラスチック製ケージ(W:310×D:360×H:175mm)を用いて1ケージあたり10匹までの群飼育とし、群分け後は、1ケージあたり5匹までの群飼育とした。飼料としては、固形飼料(MF、オリエンタル酵母工業株式会社製)を、飲料水としては、水道水をそれぞれ自由に摂取させた。
群分けは、無作為抽出法により各群の平均体重がほぼ均一になるように試験溶液の投与開始日に行った。群構成としては、偽手術群、媒体対照群、抽出物A及び抽出物W投与群の四群とした。抽出物A投与群には、投与液量として、抽出物Aの投与量がマウス1個体当たり500mg/kgとなるように、投与日の体重を基準とし、10mL/kgで算出した。同様に抽出物W投与群には、投与液量として、抽出物Wの投与量がマウス1個体当たり500mg/kgとなるように、投与日の体重を基準とし、10mL/kgで算出した。偽手術群及び媒体対照群には、0.5%(w/v)メチルセルロース溶液を10mL/kg投与した。
試験溶液の投与開始日を投与1日目として、試験溶液については1日1回投与し、投与8日目にはアミロイドβ溶液をマウスに注入した。その後、投与14日目にY字型迷路試験を実施した。各手順については後述する。
投与経路は、経口投与とした。投与方法としては、試験施設で用いられている通常の方法に従って、マウス用ディスポーザブル経口ゾンデ(有限会社フチガミ器械製)を取り付けたポリプロピレン製ディスポーザブル注射筒(テルモ株式会社製)を用いて、試験溶液を経口投与した。投与操作時には、1匹投与する毎に試験溶液を転倒混和してから、注射筒に吸引させた。なお、アミロイドβ溶液注入日には、アミロイドβ溶液注入後に試験溶液を投与し、Y字型迷路試験日には、測定の30分前に試験溶液を投与した。
マウスにペントバルビタールナトリウム(東京化成工業株式会社製)を40mg/kg腹腔内投与(投与液量:10mL/kg)することにより、マウスを麻酔した。麻酔後、頭皮にレボブピバカイン塩酸塩(ボブスカイン(登録商標)0.25%注、丸石製薬株式会社製)を皮下投与(0.1mL)した。頭皮を切開して頭蓋骨を露出させ、歯科用ドリルを用いてブレグマより側方1mm(右側)、後方0.2mmの頭蓋骨にステンレス製パイプ刺入用の穴を開けた。骨表面から2.5mmの深さまで外径0.5mmのシリコンチューブ及びマイクロシリンジに接続されたステンレス製パイプを垂直に刺入した。SCE1投与群及び媒体対照群には、脳室内にアミロイドβ溶液3μL(6nmol/3μL)をマイクロシリンジポンプで3分間かけて注入した。一方、偽手術群には注射用水3μLを同様の方法で注入した。注入後、ステンレス製パイプを挿入したまま3分間静置し、ステンレス製パイプをゆっくりと外した。その後、頭蓋穴を非吸収性骨髄止血剤(ネストップ(登録商標)、アルフレッサーファーマ株式会社製)で塞ぎ、頭皮を縫合した。
学習、記憶行動の評価方法であり、特に短期記憶の評価方法として知られている、Y字型迷路試験(例えば、非特許文献1)を実施した。試験には、1本のアームの長さが39.5cm、床の幅が4.5cm、壁の高さが12cmで、3つのアームがそれぞれ120度に分岐しているプラスチック製のY字型迷路(有限会社ユニコム製)を用いた。
評価前に、装置の床面の照度が10~40ルクスになるように調節した。評価は、試験溶液の投与後30分後に実施した。マウスをY字型迷路のいずれかのアームに置き、8分間迷路内を自由に探索させた。マウスが測定時間内に移動したアームの順番を記録し、アームに移動した回数を数え、これを総エントリー数とした。次に、この中で連続して異なる3つのアームを選択した組み合わせを調べ、この数を自発的交替行動数とした。そして、以下の式を用いて自発的交替行動率を算出した。
自発的交替行動率(%)=[自発的交替行動数/(総エントリー数-2)]×100
<抽出液の調製>
上記の抽出物Aを20%エタノールに溶解させて、固形分量30%(w/w)の抽出液Aを調製した。
抽出液A0.17gを20%エタノール1mLに溶解させてから、1mLの水を加え更に溶解させた。溶解後の溶液1mLに水9mLを加え、抽出液Aを含む溶解液を調製した(溶解液(A1))。
15mL容の遠沈管に、1ppm(w/v)のイソ吉草酸の水溶液10mLと、溶解液(A1)0.2mLとを入れ、サンプル液(1)を調製した(抽出液Aの終濃度0.017%(w/v))。
対照として、1ppm(w/v)のイソ吉草酸の水溶液10mLと、水0.2mLとの混合液を用意した(対照(1))。
0:臭いはしない
1:やっとわかる程度
2:はっきりわかる程度
3:やや強い臭い
4:強い臭い
15mL容の遠沈管に、0.1%酢酸10mLと、上述の溶解液(A1)0.2mLとを入れ、サンプル液(2)を調製した(抽出液Aの終濃度0.017%(w/v))。
対照として、0.1%酢酸10mLと、水0.2mLとの混合液を用意した(対照(2))。
サンプル液(2)及び対照(2)のそれぞれについて、試験例c1と同様の評価基準に基づき、9名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、0.1ppmのメチルメルカプタンの水溶液10mLと、上述の溶解液(A1)0.2mLとを入れ、サンプル液(3)を調製した(抽出液Aの終濃度0.017%(w/v))。
対照として、0.1ppm(w/v)のメチルメルカプタン10mLと、水0.2mLとの混合液を用意した(対照(3))。
サンプル液(3)及び対照(3)のそれぞれについて、試験例c1と同様の評価基準に基づき、9名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、1ppm(w/v)のトリメチルアミンの水溶液10mLと、上述の溶解液(A1)0.2mLとを入れ、サンプル液(4)を調製した(抽出液Aの終濃度0.017%(w/v))。
対照として、1ppm(w/v)のトリメチルアミンの水溶液10mLと、水0.2mLとの混合液を用意した(対照(4))。
サンプル液(4)及び対照(4)のそれぞれについて、試験例c1と同様の評価基準に基づき、10名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、2ppm(w/v)のジアセチルの水溶液10mLと、上述の溶解液(A1)0.2mLとを入れ、サンプル液(5)を調製した(抽出液Aの終濃度0.017%(w/v))。
対照として、2ppm(w/v)のジアセチルの水溶液10mLと、水0.2mLとの混合液を用意した(対照(5))。
サンプル液(5)及び対照(5)のそれぞれについて、試験例c1と同様の評価基準に基づき、9名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、0.01ppm(w/v)のノネナールの水溶液10mLと、上述の溶解液(A1)0.4mLとを入れ、サンプル液(6)を調製した(抽出液Aの終濃度0.034%(w/v))。
対照として、0.01ppm(w/v)のノネナールの水溶液10mLと、水0.4mLとの混合液を用意した(対照(6))。
サンプル液(6)及び対照(6)のそれぞれについて、試験例c1と同様の評価基準に基づき、11名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、0.85%(w/v)のチオグリコール酸アンモニウムの水溶液10mLと、上述の溶解液(A1)0.4mLとを入れ、サンプル液(7)を調製した(抽出液Aの終濃度0.034%(w/v))。
対照として、0.85%(w/v)のチオグリコール酸アンモニウムの水溶液10mLと、水0.4mLとの混合液を用意した(対照(7))。
サンプル液(7)及び対照(7)のそれぞれについて、試験例c1と同様の評価基準に基づき、10名のパネルにより、臭いの程度を評価した。
15mL容の遠沈管に、0.085%(w/v)のチオグリコール酸モノエタノールアミンの水溶液10mLと、上述の溶解液(A1)0.4mLとを入れ、サンプル液(8)を調製した(抽出液Aの終濃度0.034%(w/v))。
対照として、0.085%(w/v)のチオグリコール酸モノエタノールアミンの水溶液10mLと、水0.4mLとの混合液を用意した(対照(8))。
サンプル液(8)及び対照(8)のそれぞれについて、試験例c1と同様の評価基準に基づき、10名のパネルにより臭いの程度を評価した。
抽出液A0.17gを20%エタノール1mLに溶解させてから、1mLの水を加え更に溶解させた。溶解後の溶液0.3mLに水30mLを加え、抽出液Aを含む溶解液を調製した(溶解液(A2))。
5L容三角フラスコを逆さにし、火のついたタバコを三角フラスコの口の中に約5cm入れ、タバコの煙を約30~40秒間捕集した。10cm×10cmの布地(綿タオル)3枚を、煙を捕集した三角フラスコに入れて素早く密栓し、フラスコを振盪しながら布地に煙を吸わせた。5分後、タオルを取り出し、試験用布地とした。
試験用布地にスプレーで溶解液(A2)を5回スプレー(0.15ml×5回=約0.75ml)した後、よく揉み、均一にしたものをサンプル(9)とした。
サンプル(9)及び対照(9)のそれぞれについて、試験例c1と同様の評価基準に基づき、9名のパネルにより、臭いの程度を評価した。
<試験例d1:MMP-1産生阻害試験>
抽出物A及び抽出物WのMMP-1の産生阻害効果を、正常ヒト線維芽細胞に対するMMP-1の阻害作用を評価することにより調べた。
正常ヒト線維芽細胞を培養するための培地としては、5%仔牛血清含有ダルベッコ変法MEM培地(倉敷紡績株式会社製、以下、「5%FBS-DMEM培地」ともいう。)を用いた。抽出物Aを表2の濃度で5%FBS-DMEM培地に含有させた培地を調製し、これを試験サンプル含有培地とした。
試験例d1-1において、試験サンプル含有培地をハンクス緩衝液(Ca2+、Mg2+含有、HBS(+))に交換した後、4J/cm2の線量の紫外線A波(UVA)を照射してから、直ちに新鮮な試験サンプル含有培地に交換した以外は、試験例d1-1と同様の方法により、単位タンパク質量当たりのMMP-1産生量を算出した。結果を表2に示す。
試験例d1-2において、抽出物Aを抽出物Wに変更し、紫外線A波の線量を5J/cm2に変更した以外は、試験例d1-2と同様の方法により、単位タンパク質量当たりのMMP-1産生量を算出した。結果を表2に示す。
抽出物A及び抽出物Wのエラスターゼの活性阻害効果を、正常ヒト線維芽細胞由来エラスターゼの活性阻害作用を評価することにより調べた。
10cm2シャーレでコンフルエントに維持している正常ヒト線維芽細胞に対して、0.5%(V/V)のトリトンX-100緩衝液(1mmol/L PMSF、100mmol/L トリス塩酸緩衝液、pH8.0)を添加して細胞を溶解し、これを線維芽細胞由来エラスターゼの粗酵素液として用いた。エラスターゼに対する基質としては、スクシニル-L-アラニル-L-アラニル-L-アラニン p-ニトロアニリド(Suc-Ala-Ala-Ala-pNA、5mmol/L、BACHEM AG社製)を用いた。試験液として、抽出物Aをトリス緩衝液に溶解させ所定濃度の試験液を調製した。陽性対照としては、6.25mmol/LのEDTAを用いた。
エラスターゼ阻害率(%)=(1-(S’/C’))×100
抽出物Aを抽出物Wに変更した以外は、試験例d2-1と同様の方法により、抽出物Wのエラスターゼ活性阻害率を求めた。なお、陽性対照としては、12.25mmol/LのEDTAを用いた。抽出物Wにおいても、エラスターゼ活性阻害効果が認められた。
抽出物Aの線維芽細胞に対する賦活作用をMTT法により調べた。
<抽出物Aの抗糖化活性評価(試験例e1~e2)>
[試験例e1:ヒト血清アルブミンモデルにおける抗糖化活性の評価]
グルコース-ヒト血清アルブミン(HSA)の反応により生成されるAGEsに対するバガスの分解抽出物(抽出物A)の抗糖化活性(AGEs生成抑制作用)を調べた。
バガスの分解抽出物である抽出物Aを100mg/mLとなるように蒸留水で溶解し試験液原液を調製した。この試験原液を蒸留水で希釈して、0.01~100mg/mLの溶液を調製した。これを試験用のサンプルとした。陽性対照として、糖化反応阻害剤であるアミノグアニジンの水溶液(濃度3.0mg/mL)を調製した。
0.1mol/Lリン酸緩衝液(pH7.4)、8mg/mLヒト血清アルブミン(HSA、Sigma-Aldrich社製)、及び0.2mol/Lグルコース水溶液からなる反応液中に、調製した各濃度のサンプルを1/10濃度(反応終濃度)になるように添加し、60℃で40時間インキュベーションした。陰性対照としてサンプルの代わりに蒸留水を添加したものを用いた。陽性対照として上述のアミノグアニジン水溶液を用いた。なお、陽性対照に対するブランクとしてグルコースの代わりに蒸留水を添加したものを用いた。
糖化反応終了後、反応液に生成した蛍光性AGEsをマイクロプレートリーダー(SpectraMax i3、モレキュラーデバイス社)で測定した(励起波長370nm、蛍光波長440nm)。AGEsの生成阻害率(以下、単に「阻害率」ともいう)は、糖化反応においてサンプルを添加した反応液の蛍光強度をF1とし、グルコース水溶液の代わりに蒸留水を添加してインキュベーションした反応液の蛍光強度をF2とし、バガスの分解抽出物又はアミノグアニジンを添加せずにインキュベーションした反応液の蛍光強度をF3とし、ブランクとして、バガスの分解抽出物又はアミノグアニジンを添加せずに、グルコース水溶液の代わりに蒸留水を添加してインキュベーションした反応液の蛍光強度をF4として、下記の式に従って算出した。
蛍光性AGEs阻害率(%)=(1-(F1-F2)/(F3-F4))×100
次に、AGEs架橋切断試験により、バガスの分解抽出物(抽出物A)のAGEs分解活性を評価した。AGEs分解活性(AGEs架橋切断作用)は、公知の方法(例えば、Glycative Stress Research 2015年,2巻(2号),pp.58-66)である、αジケトン構造を有する1-フェニルー1,2-プロパンジオン(l-phenyl-1,2-propanedione:PPD)をモデル基質とした反応系を使用した方法で評価した。
バガスの分解抽出物である抽出物Aを20mg/mLとなるように蒸留水で溶解し、試験用のサンプルを調製した。
0.16mol/Lのリン酸緩衝液(pH7.4)、2mmol/mLのPPDの組成の反応液中に、上記で調製したサンプルを1/2濃度(10mg/mL)になるように添加し、37℃で8時間インキュベーションした。陰性対照としてはサンプルの代わりに蒸留水を添加したものを用いた。陽性対照としてPTB(N-phenacylthiazoliumbromide)を用いた。反応液を20℃、3000×gで10分間遠心分離し、上清を得た。上清中の安息香酸量を逆相HPLCで分析した。反応液中の安息香酸量は、別途測定したサンプル中の安息香酸量を差し引いて求めた。1molのPPDは1molの安息香酸を生成することから、以下の式で架橋切断率を算出した。
架橋切断率(%)={(A-B)/C}×100
A:反応液中の安息香酸量
B:サンプル中の安息香酸量
C:反応に供したPPD量(基質量)
サンプル及びPTB溶液(5mmol/L)における架橋切断率と、PTB(5mmol/L)を100%としたときのサンプルにおける架橋切断率の値(切断率相対値)を求めたところ、サンプルの架橋切断率は8.50、PTBの架橋切断率は20.1であり、サンプルの切断率相対値は42.29%であった。よって、抽出物AがAGEsの分解活性を有していることが示された。
[試験例e3:ヒト血清アルブミンモデルにおける抗糖化活性の評価]
バガスの分解抽出物として上述の抽出物Wを用いたこと、サンプルの調製において蒸留水ではなくジメチルスルホキシド(DMSO)を用いたこと以外は、試験例e1と同様にして、抗糖化活性を評価した。陽性対照としては、アミノグアニジンの水溶液(最終濃度0.3mg/mL)を用いた。陽性対照であるアミノグアニジンの0.3mg/mLにおける蛍光性AGEs(HSA)の阻害率は74.6±0.8%であった。測定結果を図7に示す。また、抽出物Wの各濃度におけるサンプルの阻害率から算出したIC50(50%生成阻害濃度)は、0.14mg/mLで、抽出物Wは、抗糖化活性を有していることが示された。
(サンプル調製)
バガスの分解抽出物である抽出物Wを50%DMSOで溶解し、20mg/mL溶液を調製した。この溶液を50%DMSOで段階希釈して、試験用のサンプルとした。陽性対照としては、10mmol/LのPTB(N-phenacylthiazoliumbromide)溶液を用いた。
試験溶液又はPTB溶液(10mmol/L)と、10mmol/LのPPD溶液と、0.2mol/Lリン酸緩衝液(pH7.4)と、を5:1:4の割合で混合し、37℃で8時間反応させた(n=3)。反応終了後、塩酸を加えて反応停止させた。その後、反応液を20℃で3,000×gで10分間遠心分離し、上清中の安息香酸量を逆相HPLCで分析した。反応液中の安息香酸量は、別途測定したサンプル中の安息香酸量を差し引いて求めた。1molのPPDは1molの安息香酸を生成することから、以下の式で架橋切断率を算出した。架橋切断の相対値(切断率相対値)はPTBの架橋切断率を100としたときの、各濃度の架橋切断率の値(%)である。なお、測定装置としては、島津超高速液体クロマトグラフNexeraシステム(株式会社島津製作所製)を用いた。
架橋切断率(%)={(A-B)/C}×100
A:反応液中の安息香酸量
B:サンプル中の安息香酸量
C:反応に供したPPD量(基質量)
サンプル及びPTB溶液(5mmol/L)における架橋切断率と、PTB(5mmol/L)を100%としたときのサンプルにおける架橋切断率の値(切断率相対値)を求めたところ、サンプルの架橋切断率は10.07、PTBの架橋切断率は22.4であり、サンプルの切断率相対値は44.87%であった。よって、抽出物WがAGEsの分解活性を有していることが示された。
<試験f1:抽出物AによるRBL-2H3細胞脱顆粒抑制試験>
[試験液の調製]
抽出物Aを水に溶解させて、50mg/mLの試験液原液を調製した。この試験液原液を以下の表5に示す緩衝溶液で希釈し、検体濃度2000、1000及び500μg/mLの試験液をそれぞれ調製した。
ラット好塩基球性白血病細胞であるRBL-2H3細胞(国立研究開発法人 医薬基盤・健康・栄養研究所)を96ウェルプレートに播種後、一晩培養した。表5に示す組成を有し、更に抗DNP-IgE抗体を含む培地を添加し、37℃で2時間反応させた後、細胞を緩衝溶液で洗浄した。さらに、調製した2000、1000及び500μg/mLの試験液を、終濃度がそれぞれ1000μg/mL(実施例f1)、500μg/mL(実施例f2)及び250μg/mL(実施例f3)となるように添加した。その後、37℃で10分間反応させてから、DNP標識ヒト血清アルブミンを加え、37℃で更に3時間反応させた。また、試験液を添加せず、緩衝溶液のみを添加したものを未処置対照(比較例f1)、ウォルトマンニン(和光純薬株式会社)を終濃度25nmol/Lとなるように添加したものを陽性対照として同様に試験を行った。また、抗DNP-IgE抗体を含まない培地を添加した後、緩衝溶液及びDNP標識ヒト血清アルブミンを順次加えて、同様に反応させたものを抗原未刺激対照とした。
実施例f1~f3、比較例f1、陽性対照、抗原未刺激対照及びサンプルブランクの各サンプルについて、マイクロプレートリーダー(SpectraMax M2e、モレキュラーデバイス社)を用いて、顆粒中に存在するβ-ヘキソサミニダーゼと基質との反応により生じたp-ニトロフェノールの吸光度を測定した(測定波長:405nm、対照波長:650nm)。
放出率(%)=細胞上清側の吸光度/(細胞上清側の吸光度+細胞溶液側の吸光度)
脱顆粒率(%)={(試験液の放出率-抗原未刺激対照の放出率)/(未処置対照の放出率-抗原未刺激対照の放出率)の平均値}×100
[試験液の調製]
抽出物Wをエタノールに溶解させて、50mg/mLの試験液原液を調製した。この試験液原液を上述の表5に示す緩衝溶液で希釈し、検体濃度1000、500及び250μg/mLの試験液をそれぞれ調製した。
調製した抽出物Wを含む試験液を用い、上述の試験例f1における試験操作中、試験液の終濃度が500μg/mL(実施例f4)、250μg/mL(実施例f5)及び125μg/mL(実施例f6)となるように試験液を添加した以外は、試験例1と同様の方法によって脱顆粒率を算出した。比較例f1は、試験例f1と同様の未処置対照である。
<アンジオテンシン変換酵素阻害試験>
Nakanoらの方法(Nakano et al, Biosci.Biotechnol. Biochem., 70, 1118-1126(2006))に基づき、アンジオテンシン変換酵素阻害試験を行った。
抽出物A及び抽出物Wそれぞれ1.0gを、50%(V/V)エタノール溶液20mlで抽出後、0.1mol/Lのヘペス緩衝液(pH8.3)にて適宜希釈し、表6に示す濃度の試験液を調製した。0.1mol/Lのヘペス緩衝液(未処置区)又は試験液を96穴マイクロプレートに25μLずつ加え、更に20mU/mLのACE溶液を25μL加えて37℃で5分間インキュベートした。ここに8mmol/Lの基質(ヒプリル-L-ヒスチジル-L-ロイシン;Hip-His-Leu)溶液を25μL更に加え、37℃で30分間反応させた。その後、0.1mol/Lの水酸化ナトリウム水溶液を25μL加えて反応を停止させ、1質量%のオルトフタルアルデヒド(OPA)水溶液25μLを更に加えて20分間放置した。その後、0.1mol/Lの塩酸を25μL加えて測定用検体とした。測定用検体を室温で10分間放置し、マイクロプレートリーダーを用いて以下の条件により蛍光強度を測定した。なお、ブランクにはACE溶液の代わりにリン酸緩衝生理食塩水を用いた。
機種:SpectraMax M2e(モレキュラーデバイス社製)
測定条件:蛍光、endpointモード、ボトムリード
励起波長:355nm
蛍光波長:460nm
<試験例h1:抽出物Aによる風味向上試験>
[豆乳]
豆乳(商品名:スゴイダイズ無調整タイプ、大塚チルド食品株式会社製)100gに、抽出物Aを表7に示す濃度になるように、抽出物Aの溶液(固形分濃度0.3%)を0.6g(実施例h1-1)又は0.3g(実施例h1-2)添加して、被検品(実施例h1-1、h1-2)を調製した。
米酢50g、水200g、及びグラニュー糖15gの酢含有飲料に、抽出物Aの溶液(固形分濃度0.3%)を1.7g添加して被検品(実施例h1-3)を調製した。
牛肉30gと、抽出物Aの溶液(固形分濃度0.3%)0.012gとを、フードプロセッサーにより牛肉をミンチ状にしながら混合した。このミンチ肉を10gずつアルミカップに小分けして、200℃のオーブンで5分間焼成し、上下を返して更に5分間焼成した。焼成後人肌程度に冷却したものを被検品(実施例h1-4)とした。
鶏胸ひき肉25gと、水で戻した大豆たんぱく質(商品名:ニューソイミーS 20F、日清オイリオグループ株式会社製)10gと、抽出物Aの溶液(固形分濃度0.3%)0.1gとを混合した。これを一口大に丸めて沸騰水で5分間加熱して調製した鶏団子を被検品(実施例h1-5)とした。
各実施例の飲食品に対し、表7~表10に示す項目について官能評価を実施した。なお、官能評価の評価基準は対照品(抽出物Aを含まないもの)と比較した相対評価とし、対照品を0点とした場合の各実施例の評価を-2点~2点の間の数値で評価した。各評価項目について、-2点が「対照品と比較して最もその風味を感じない」ことを意味し、2点が「対照品と比較して最もその風味を感じる」ことを意味する。評価点の平均値を表7~表10に示す。なお、表中の「固形分濃度」は、被検品全量基準における抽出物Aの固形分濃度を示し、以下において特記しない場合には同様である。
[豆乳、酢含有飲料、牛肉加工品(肉団子)]
試験例h1において、抽出物Aを抽出物Wに変更した以外は試験例h1と同様に被検品(実施例h2-1~h2-3)を調製した。
8%食塩水500gに、抽出物Wの溶液(固形分濃度0.3%)を4.175g添加した浸漬液を調製した。冷凍切身のサバを解凍後、皮に切れ目を入れてから浸漬液に10分間浸漬させた。グリルによりサバの両面を7分間ずつ焼成し、被検品(実施例h2-4)を調製した。被検品は、一旦冷凍したものを電子レンジで加熱してから官能評価に供した。
市販飲料(商品名:ミルミル、株式会社ヤクルト製)100質量部、10%酢酸0.275質量部、アルパルテーム(味の素株式会社製)0.007質量部、及びβ-カロチンの1%溶液(三菱化学フーズ株式会社製)0.015質量部を含む乳酸菌飲料を調製した。この乳酸菌飲料100gに、抽出物Wの溶液(固形分濃度0.3%)を0.075g添加して被検品(実施例h2-5)を調製した。
[甘蔗由来のエキスの製造]
原料である甘蔗汁(原料糖製造工場の製糖工程にて得られた石灰清浄後の清浄汁、沖縄産、固形分14%)7500Lを、カートリッジフィルター(アドバンテック株式会社製、コットンワインドカートリッジフィルター、TCW-10-CSD型)で濾過処理し、清浄汁ろ過処理物を得た。合成吸着剤(三菱ケミカル株式会社製、SP-207)500Lを樹脂塔(内径800mm、高さ2,000mm)に充填し、これに上記の清浄汁ろ過処理物を、流速2500L/時間(SV=5.0(時間-1))で通液した。溶出パターンを図10に示す。図1の(A)が通液開始点である。なお、清浄汁通過中は、ウォータージャケットには、80℃の水を常に循環させた。
樹脂塔から溶出した画分のうち後半の1460L(図10においてaの部分)を、遠心式薄膜真空蒸発装置(株式会社大川原製作所、エバポールCEP-5S)にて約50倍の濃度に減圧濃縮したのち、一晩凍結乾燥して、茶褐色の粉末(I)8.4kgを得た。この粉末をエタノール及び水に溶解させて、固形分濃度30質量%の甘蔗由来のエキス(以下、「甘蔗エキス」ともいう。)を調製した。
対照品は、上述の甘蔗エキスの希釈液(固形分濃度0.3%)を使用して、表16に記載の方法で調製した。一方、実施例h3-1~h3-4、及び実施例h3-7~h3-8(豆乳、酢含有飲料及び焼サバ)については、試験例h1又は試験例h2と同様の方法により被検品を調製した。実施例h3-5~h3-6及び実施例h3-9~h3-10(肉団子及び乳酸菌飲料)については、表16に記載の対照品の調製方法において、甘蔗エキスの希釈液を抽出物Aの溶液又は抽出物Wの溶液(固形分濃度0.3%)にそれぞれ変更して被検品を調製した。
上述の抽出物A及び甘蔗エキスを0.2%酢酸水溶液に添加して、最終固形分濃度が45質量ppm、90質量ppm及び180質量ppmである0.2%酢酸水溶液(実施例h4-1~h4-3、比較例h4-1~h4-3)を調製した。この酢酸溶液を、味認識装置(TS-5000Z、インテリジェントセンサーテクノロジー社製)を用いて、酸味度を測定した。当該味認識装置にて測定する際には、酸味センサー(CAO)の先味(相対値)により測定した。抽出物A及び甘蔗エキスを添加していない酢酸水溶液(対照)の酸味度を0とした、各酢酸水溶液の酸味度の相対値を表22に示す。酸味度の相対値が0以下であることは、対照である0.2%酢酸水溶液と比較して、酢酸により感じる嫌味(酸味)が低減されたことを示す。
[材料]
以下の試験例では、下記に示す材料を用いた。
(細胞)
マウス筋芽細胞C2C12細胞(ATCC、CRL-1772)
増殖培地の組成:DMEM培地、10%FBS、抗生物質添加
分化培地の組成:DMEM培地、0.5%FBS、抗生物質添加
ダルベッコ改変イーグル培地(DMEM培地、ナカライテスク株式会社)
Fetal Bovine Serum(FBS)(Cell CultureBioscience社)
ペニシリン-ストレプトマイシン混合溶液(ナカライテスク株式会社)
0.25%トリプシン/EDTA混合溶液(ナカライテスク株式会社)
ダルベッコPBS(-)(日水製薬株式会社)
ゼラチン(Aタイプ、MP Biomedicals社)
ヘキスト(核染色試薬、ヘキスト 33342溶液、株式会社同仁化学研究所)
ミトトラッカー(MitoTracker Mitochondrion-Selective Probes、インビトロジェン社)
ローダミン(VectaCell Rhodamine 123、フナコシ株式会社)
4%-パラホルムアルデヒド・リン酸緩衝液(ナカライテスク株式会社)
一次抗体(Anti-Myosin Heavy Chain Purified clone:MF20、eBioscience社)
二次抗体(Alexa Fluor 555 F(ab)2 fragment of goat anti-mouse IgG (H+L)、ライフテクノロジーズジャパン株式会社)
以下の試験例において使用する細胞の前培養を行った。
C2C12細胞を、増殖培地を用いてT-75フラスコ(75cm2U字型カントネック細胞培養用フラスコ、コーニング社)にて起眠させた。T-75フラスコをCO2インキュベーター(5%CO2、37℃、湿潤)内に入れ、C2C12細胞を培養した。培地交換は一日おきに行い、80%コンフルエントに到達した時点で細胞を回収し、これを試験に用いた。なお、以下の試験において、C2C12細胞を培養する際のウェルプレートには、下記に示すコーティング方法によりゼラチンをコートしたプレートを使用した。
(1)0.75%ゼラチン水溶液をオートクレーブにて滅菌処理した。
(2)プレート内の各ウェルに、100μLの0.75%ゼラチン水溶液を添加し、このウェルをCO2インキュベーター(5%CO2、37℃)内で2時間静置した。
(3)0.75%ゼラチン水溶液を除去し、得られたプレートを使用した。
増殖培地を用いて、4×104セル/0.1mL/ウェルで、蛍光観察用96ウェルプレート(オプティカルボトムプレート、Nunc社)に前培養した細胞を播種した。このウェルプレートを、CO2インキュベーター内(5%CO2、37℃)で2日間培養した。その後、培地を分化培地に交換して、4日間培養することにより、筋管細胞を形成させた。筋管細胞を形成後、抽出物Aを含む分化培地(実施例i1-1~i1-3)、抽出物Wと1%(v/v)エタノールを含む分化培地(実施例i1-4~i1-6)、抽出物を含まない分化培地(比較例i1-1)、抽出物を含まず1%(v/v)エタノールを含む分化培地(比較例i1-2)、又は50μMの濃度のレスベラトロールと1%(v/v)エタノールを含む分化培地(陽性対照i1-1)に交換し、それぞれ培養した。培養を開始してから48時間後に、培養を終了した。これらを、実施例i1-1~i1-6、比較例i1-1~i1-2、又は陽性対照i1-1の試験サンプルとした。なお、実施例i1-1~i1-6において、バガスの分解抽出物の分化培地中における終濃度を表23に示す。
試験例i1-1と同様の方法により、筋管細胞を培養した。これらを、実施例i1-7~i1-11、比較例i1-3~i1-4、又は陽性対照i1-2の試験サンプルとした。なお、実施例i1-7~i1-11において、バガスの分解抽出物の分化培地中における終濃度を表24に示す。その後、培養上清を除去した後、5μg/mLヘキスト(核染色用試薬)、及び500nMミトトラッカー(ミトコンドリア染色試薬)を含む分化培地をウェルプレートに添加し、試験例i1-1と同様の方法により蛍光強度を測定した。ミトトラッカーを含む分化培地の蛍光強度を、ヘキストを含む分化培地の蛍光強度で除することにより、細胞当たりのミトコンドリア量を算出した。比較例i1-3の試験サンプルにおけるミトコンドリア量を100%としたときの実施例i1-7~i1-8の試験サンプルにおける相対値(%)を求めた。また、比較例i1-4の試験サンプルにおけるミトコンドリア活性を100%としたときの実施例i1-9~i1-11及び陽性対照i1-2の試験サンプルにおける相対値(%)を求めた。試験は5回ずつ実施し、平均値を表24に示す。
増殖培地を用いて、4×104セル/0.1mL/ウェルで、蛍光観察用96ウェルプレートに前培養した細胞を播種した。このウェルプレートを、CO2インキュベーター内(5%CO2、37℃)で2日間培養した。その後、抽出物Aを含む分化培地(実施例i2-1~i2-2)、抽出物を含まない分化培地(比較例i2-1)、抽出物を含まず0.1%(v/v)DMSOを含む分化培地(比較例i2-2)、又は0.5μM LDN-193189(4-(6-(4-(ピペラジン-1-イル)フェニル)ピラゾロ[1,5-a]ピリミジン-3-イル)キノリン)と0.1%(v/v)DMSOを含む分化培地(陽性対照i2)に交換した。培地交換後、3日間培養して、固定後に抗ミオシン重鎖(MHC)抗体にて免疫染色を行った。なお、実施例i2-1~i2-2において、バガスの分解抽出物の分化培地中における終濃度を表25に示す。
Fusion index(% of MHC+ nuclei)=MHC陽性核数/総核数×100
<試験例j1:骨芽細胞分化促進試験>
[材料]
試験例j1では、下記に示す材料を用いた。
(細胞)
マウス頭蓋冠由来細胞MC3T3-E1(理研セルバンク、RCB1126)
α-MEM培地、10%FBS、抗生物質添加
α-MEM培地(フェノールレッドフリー、製品番号41061-029、インビトロジェン社)
ペニシリン-ストレプトマイシン混合溶液(製品番号26253-84、ナカライテスク株式会社)
0.25%トリプシン-EDTA混合溶液(製品番号32777-44、ナカライテスク株式会社)
ダルベッコPBS(-)(製品番号05913、日水製薬株式会社)
アルカリホスファターゼ活性測定キット(LabAssay ALP、製品番号291-58601、和光純薬工業株式会社)
タンパク質質量測定キット(Micro BCA Protein Assay Reagent Kit、製品番号23235、PIERCE社)
細胞溶解・タンパク質抽出試薬(Cell-LyEX1、製品番号300-34761、和光純薬工業株式会社)
10%中性緩衝ホルマリン液(製品番号062-01661、和光純薬工業株式会社)
Calcein AM(製品番号PK-CA707-80011、PromoKine社)
組換え骨形成タンパク質(Bone Morphogenetic Protein-2(BMP-2)、R&DSystems社)
MC3T3-E1細胞を、増殖培地を用いてT-75フラスコ(75cm2U字型カントネック細胞培養用フラスコ、コーニング社)にて起眠させた。T-75フラスコをCO2インキュベーター(5%CO2、37℃、湿潤)内に入れ、C2C12細胞を培養した。培地交換は一日おきに行い、80%コンフルエントに到達した時点で細胞を回収し、これを試験に用いた。
MC3T3-E1細胞における骨芽細胞分化促進を、骨芽細胞の分化マーカーの1つであるアルカリホスファターゼ(ALP)活性を指標として確認した。
前培養したMC3T3-E1細胞を1.2×105セル/0.2mL/ウェルとなるよう培地で調整し、48ウェルプレートに播種した。翌日、100μg/mLの抽出物A含む培地(実施例j1-1)、100μg/mLの抽出物Wと1%(v/v)エタノールを含む培地(実施例j1-2)、抽出物を含まない培地(比較例j1-1)、抽出物を含まず、1%(v/v)エタノールを含む培地(比較例j1-2)、あるいは、BMP-2を含有する培地(陽性対照j1)に置換し、それぞれ7日間、14日間、及び21日間培養した。各日数培養後に、細胞をPBSで1回洗浄し、プレートごと冷凍保存した。培地は、3~4日間毎に交換した。
[材料]
試験例j2では、下記に示す材料を用いた。
(細胞)
ヒト破骨前駆細胞(コスモバイオ株式会社 PT-267 Lot.RBW-F-OSH-HBV)
ヒト破骨細胞培養用メディウム、OSCMHB、コスモバイオ株式会社
メラトニン(M5250、Sigma-Aldrich社)
TRAP染色キット(AK04F、PMC社)
上記の培地を用いて、抽出物Aの250μg/mL溶液を調製し、これを試験液とした(実施例j2-1)。
ヒト破骨前駆細胞を96ウェルの培養プレートに、約0.3×105cells/50μl/ウェルとなるように播種した。ここに試験液を50μlL/ウェル添加して、37℃、5%CO2下の条件で7日間培養した。TRAP染色キットを用いて培養した細胞をTRAP染色し、顕微鏡による観察を行った。同様に、試験液を添加しない培地(比較例j2-1)、陽性対照としてメラトニンを1000μM含む培地(陽性対照j2)においても、同様に試験を行った。顕微鏡の観察結果を図11に示す。図11(a)が実施例j2-1、図11(b)が比較例j2-1、図11(c)が陽性対照j2の観察結果である。
破骨細胞は成熟すると細胞同士が融合し、多核化することが知られている。そこで、破骨細胞の単核細胞の割合を測定し、破骨細胞分化抑制効果を確認した。
Claims (88)
- バガスの分解抽出物を有効成分として含有する、肥満抑制剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項1に記載の肥満抑制剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項2に記載の肥満抑制剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項3に記載の肥満抑制剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項3に記載の肥満抑制剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項4又は5に記載の肥満抑制剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項2に記載の肥満抑制剤。 - 脂肪蓄積抑制作用に基づくものである、請求項1~7のいずれか一項に記載の肥満抑制剤。
- バガスの分解抽出物を有効成分として含有する、脂肪蓄積抑制剤。
- バガスの分解抽出物を有効成分として含有する、抗認知症剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項10に記載の抗認知症剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項11に記載の抗認知症剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項12に記載の抗認知症剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項12に記載の抗認知症剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項13又は14に記載の抗認知症剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項11に記載の抗認知症剤。 - バガスの分解抽出物を有効成分として含有する、短期記憶障害改善/抑制剤。
- バガスの分解抽出物を有効成分として含有する、消臭剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理及び亜臨界水処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項18に記載の消臭剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項19に記載の消臭剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項20に記載の消臭剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項20に記載の消臭剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項21又は22に記載の消臭剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項19に記載の消臭剤。 - バガスの分解抽出物を有効成分として含有する、抗老化剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項25に記載の抗老化剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項26に記載の抗老化剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項27に記載の抗老化剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項27に記載の抗老化剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項28又は29に記載の抗老化剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項26に記載の抗老化剤。 - バガスの分解抽出物を有効成分として含有する、細胞外マトリックス分解酵素阻害剤。
- バガスの分解抽出物を有効成分として含有する、線維芽細胞賦活剤。
- バガスの分解抽出物を有効成分として含む、抗糖化剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項34に記載の抗糖化剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項35に記載の抗糖化剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項36に記載の抗糖化剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項36に記載の抗糖化剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項37又は38に記載の抗糖化剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項35に記載の抗糖化剤。 - 請求項34~40のいずれか一項に記載の抗糖化剤を含有する、抗糖化用飲食品。
- バガスの分解抽出物を有効成分として含有する、抗I型アレルギー剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項42に記載の抗I型アレルギー剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項43に記載の抗I型アレルギー剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項44に記載の抗I型アレルギー剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項44に記載の抗I型アレルギー剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項45又は46に記載の抗I型アレルギー剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項43に記載のI型アレルギー剤。 - 肥満細胞又は好塩基球の脱顆粒抑制作用に基づくものである、請求項42~48のいずれか一項に記載の抗I型アレルギー剤。
- バガスの分解抽出物を有効成分として含有する、肥満細胞又は好塩基球の脱顆粒抑制剤。
- バガスの分解抽出物を有効成分として含有する、抗高血圧剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項51に記載の抗高血圧剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項52に記載の抗高血圧剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項53に記載の抗高血圧剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項53に記載の抗高血圧剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項54又は55に記載の抗高血圧剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項52に記載の抗高血圧剤。 - バガスの分解抽出物を有効成分として含有する、アンジオテンシン変換酵素阻害剤。
- バガスの分解抽出物を含有する、風味向上剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項59に記載の風味向上剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項60に記載の風味向上剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項61に記載の風味向上剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項61に記載の風味向上剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項62又は63に記載の風味向上剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項60に記載の風味向上剤。 - 飲食品の好ましい風味を増強する、請求項59~65のいずれか一項に記載の風味向上剤。
- 飲食品の嫌味を低減する、請求項59~66のいずれか一項に記載の風味向上剤。
- 請求項59~67のいずれか一項に記載の風味向上剤を含有する、飲食品。
- バガスの分解抽出物を含有する、飲食品の好ましい風味増強剤。
- バガスの分解抽出物を含有する、飲食品の嫌味低減剤。
- バガスの分解抽出物を有効成分として含有する、筋肉増強剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項71に記載の筋肉増強剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項72に記載の筋肉増強剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項73に記載の筋肉増強剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項73に記載の筋肉増強剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項74又は75に記載の筋肉増強剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項72に記載の筋肉増強剤。 - バガスの分解抽出物を有効成分として含有する、筋管細胞分化促進剤。
- バガスの分解抽出物を有効成分として含有する、ミトコンドリア賦活剤。
- バガスの分解抽出物を有効成分として含有する、骨代謝改善剤。
- 前記バガスの分解抽出物は、アルカリ処理、水熱処理、酸処理、亜臨界水処理及び爆砕処理からなる群より選ばれる少なくとも1種の分解処理により得られる分解処理液である、請求項80に記載の骨代謝改善剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体を充填したカラムに通液することより得られる画分である、請求項81に記載の骨代謝改善剤。
- 前記固定担体は、合成吸着剤又はイオン交換樹脂である、請求項82に記載の骨代謝改善剤。
- 前記固定担体が合成吸着剤であり、前記バガスの分解抽出物は、該合成吸着剤に吸着された成分を、水、メタノール、エタノール及びこれらの混合物からなる群より選ばれる少なくとも1種の溶媒で溶出させることにより得られる画分である、請求項82に記載の骨代謝改善剤。
- 前記合成吸着剤は、芳香族系樹脂、アクリル酸系メタクリル樹脂、又はアクリロニトリル脂肪族系樹脂である、請求項83又は84に記載の骨代謝改善剤。
- 前記バガスの分解抽出物は、前記分解処理液を、固定担体としての合成吸着剤を充填したカラムに通液し、該合成吸着剤に吸着された成分を、エタノール及び水の混合溶媒で溶出させて得られる画分であり、
前記合成吸着剤は、無置換基型の芳香族系樹脂であり、
前記カラムの温度は20~60℃であり、
前記混合溶媒のエタノール及び水の体積比(エタノール/水)は50/50~60/40である、請求項81に記載の骨代謝改善剤。 - バガスの分解抽出物を有効成分として含有する、骨形成促進剤。
- バガスの分解抽出物を有効成分として含有する、骨吸収抑制剤。
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