WO2023191310A1 - Composition d'amélioration de la fonction antioxydante, anti-vieillissement, antibactérienne, anti-inflammatoire ou cognitive contenant un extrait de marc de schizandra chinensis baillon fermenté en phase solide - Google Patents

Composition d'amélioration de la fonction antioxydante, anti-vieillissement, antibactérienne, anti-inflammatoire ou cognitive contenant un extrait de marc de schizandra chinensis baillon fermenté en phase solide Download PDF

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WO2023191310A1
WO2023191310A1 PCT/KR2023/002343 KR2023002343W WO2023191310A1 WO 2023191310 A1 WO2023191310 A1 WO 2023191310A1 KR 2023002343 W KR2023002343 W KR 2023002343W WO 2023191310 A1 WO2023191310 A1 WO 2023191310A1
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schisandra chinensis
solid
extract
schisandra
antibacterial
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English (en)
Korean (ko)
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정유진
권수연
정유민
황가은
박종목
정서현
임보규
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한국화학연구원
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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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/79—Schisandraceae (Schisandra family)
    • 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]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/005—Antimicrobial preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine

Definitions

  • the present invention relates to compositions for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement.
  • the present invention relates to a method for producing a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement.
  • Schizandra chinensis Baillon is a red fruit with a harmonious combination of five tastes: sour, sweet, spicy, bitter, and salty.
  • the main functional ingredient contained in Schizandra chinensis is an intermediate substance in the process of lignin synthesis. It is composed of lignans.
  • Lignan is the main component of Schisandra chinensis' medicinal properties and is composed of Schisandrin, Gomisin, etc. Lignans are known to have antioxidant, antibacterial, and anticonvulsant effects (reduce GABA enzymes), and can prevent neurodegeneration and dementia such as Alzheimer's.
  • the lignan content of Schisandra chinensis is 10 to 30 times higher than that of nuts, and 80% is present in Schisandra chinensis seeds and 20% in the pulp.
  • Schisandra fruit is a fruit with a very high organic acid content and has difficulty in natural fermentation, so it is widely used as tea extracted by leaching the fruit with alcohol or pickling it.
  • it is difficult to extract Schisandra chinensis lignans, and it is not good for health due to the excessive use of sugar.
  • the lignans present in Schisandra seeds cannot be consumed, so a trace amount of lignans is consumed, which is effective. There is a problem that it is difficult to see.
  • Schisandra chinensis products are mainly manufactured by processing juice, fruit extract, or fermented extract, and after processing, Schisandra chinensis (pulp, skin, and seeds discarded after extraction) remains as a by-product of processing.
  • Schisandra chinensis pulp, skin, and seeds discarded after extraction
  • the seed part with the highest lignan content does not contribute as an ingredient in the product and is discarded in large quantities without any specific use.
  • Schisandra chinensis a by-product of Schisandra chinensis, the medicinal plant with the largest production volume in Korea, is classified as general waste and is 100% landfilled or incinerated. Environmental problems caused by carbon dioxide (CO 2 ) generated during the treatment process cannot be ignored.
  • CO 2 carbon dioxide
  • the present inventors performed solid-state fermentation of Schisandra chinensis with mushrooms, etc. for up-cycling of Schisandra chinensis, a food by-product.
  • Schisandra chinensis is subjected to solid-state fermentation using mushrooms, etc.
  • the content of lignans and GABA ( ⁇ -aminobutyric acid) increases. This shows the transformation of Schisandra chinensis, which was a waste material, into a new functional material.
  • the purpose of the present invention is to provide a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement.
  • the purpose of the present invention is to provide a method for producing a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement.
  • composition of 1 above, wherein the extract is an ethanol extract, and is an antioxidant, anti-aging, antibacterial, anti-inflammatory, or cognitive function improvement composition.
  • composition according to item 1 above, wherein the extract is an ultrasonic extract, an antioxidant, anti-aging, antibacterial, anti-inflammatory, or improving cognitive function.
  • a method for producing a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement comprising the step of extracting solid-state fermented Schisandra chinensis cucumber.
  • a manufacturing method further comprising:
  • composition of the present invention has a DPPH radical scavenging effect, an antibacterial effect against acne bacteria, a NO (nitric oxide) inhibitory effect, and an AChE (aetylcolinesterase) inhibitory effect, so it can help improve antioxidant, anti-aging, antibacterial, anti-inflammatory, or cognitive functions. there is.
  • Figure 1 shows six types of useful microorganism strains used in solid-state fermentation.
  • Figure 2 shows the pre-cultivation (seed culture) and main culture processes of six types of useful microbial strains used in solid-state fermentation.
  • Figure 3 shows the solid-state fermentation process of Schisandra chinensis and Schisandra chinensis meal.
  • Figure 4 is a photograph of solid-state fermentation of Schisandra chinensis using six types of useful microorganisms.
  • Figure 5 is a photograph of Schisandra chinensis cucumbers subjected to solid-state fermentation using six types of useful microorganisms.
  • Figure 6 shows the manufacturing process of Schisandra chinensis extract.
  • Figure 7 shows the manufacturing process of solid-state fermented Schisandra chinensis extract or solid-state fermented Schisandra chinensis extract.
  • Figure 8 is a graph showing the particle size distribution of Schisandra chinensis cucumber samples with small particle sizes measured with a micro particle size analyzer.
  • Figure 9 is a photograph of Schisandra chinensis cucumber samples with large particle sizes classified into three groups.
  • Figure 10 is a graph showing the particle size distribution of sample 3 of Figure 9 measured with a micro particle size analyzer.
  • Figure 11 shows the results of quantitative analysis of the active ingredient (lignan) of the extract obtained by heat-extracting solid-state fermented Schisandra chinensis with 100% distilled water through MS (mass spectrometry) analysis.
  • Figure 12 shows the results of quantitative analysis of the active ingredient (lignan) of the extract obtained by ultrasonic extraction of solid-state fermented Schisandra chinensis with 100% distilled water through MS analysis.
  • Figure 13(a-c) shows the results of quantitative analysis of the active ingredient (lignan) of the extract obtained by ultrasonic extraction of Schisandra chinensis root with 60% ethanol using solid-state fermentation using MS analysis.
  • Figure 14 is a photograph showing the process of fractionating the extract obtained by ultrasonic extraction of solid-state fermented Schisandra chinensis with 60% ethanol.
  • Figure 15 (a-c) shows the results of analyzing the active ingredients of the fraction of the extract obtained by ultrasonic extraction of solid-state fermented Schisandra chinensis with 60% ethanol.
  • Figures 16a and 16b show the results of lignan quantitative analysis of the extract obtained by ultrasonic extraction of solid-phase fermented Schisandra chinensis with 60% ethanol using HPLC analysis.
  • Figure 17 shows the cytotoxicity test results of ethanol, an extraction solvent.
  • Figure 18 shows the results of a cytotoxicity test of extracts obtained by ultrasonic extraction of Schisandra chinensis, Schisandra chinensis, solid-state fermented Schisandra chinensis, or solid-state fermented Schisandra chinensis with 60% ethanol.
  • Figure 19 shows the results of tests for antioxidant and anti-aging efficacy of extracts obtained by ultrasonic extraction of Schisandra chinensis, Schisandra chinensis, solid-state fermented Schisandra chinensis, or solid-state fermented Schisandra chinensis with 60% ethanol.
  • Figure 20 shows the antibacterial test results of the extract obtained by ultrasonic extraction of solid-state fermented Schisandra chinensis or solid-state fermented Schisandra chinensis peel with 60% ethanol.
  • Figures 21 and 22 show the anti-inflammatory test results of extracts obtained by ultrasonic extraction of Schisandra chinensis, Schisandra chinensis, solid-state fermented Schisandra chinensis or solid-state fermented Schisandra chinensis with 60% ethanol, showing the nitrite concentration ( Figure 21) and nitrogen monoxide ( Nitric Oxide) production reduction rate ( Figure 22).
  • Figure 23 shows the results of a cognitive function improvement test of extracts obtained by ultrasonic extraction of Schisandra chinensis, Schisandra chinensis fruit, solid-state fermented Schisandra chinensis, or solid-state fermented Schisandra chinensis fruit with 60% ethanol.
  • the present invention provides a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement comprising solid-state fermented Schisandra chinensis extract or fractions thereof.
  • Schisandra gourd is a processing by-product of pulp, skin, and seeds that are discarded after extracting Schisandra fruit.
  • Schisandra gourd contains lignans as active ingredients.
  • the lignan may be Schizandrin, Gomisin A, Gomisin N, etc.
  • Lignans are contained in large quantities in the seeds of Schisandra chinensis compared to the pulp or skin.
  • Schisandra gourd extract has a higher lignan content than Schisandra chinensis extract.
  • Schisandra chinensis extract when using Schisandra chinensis extract, it has superior DPPH radical scavenging ability, antibacterial activity, and AChE (aetylcolinesterase) inhibitory ability compared to Schisandra chinensis extract.
  • the solid-state fermented Schisandra chinensis fruit extract of the present invention may be an extract of solid-state fermented Schisandra chinensis marmalade obtained by solid-state fermentation of crushed Schisandra chinensis fruit.
  • crushed Schisandra gourd seeds containing a large amount of lignans are also crushed, so the extract may contain more lignans.
  • Schisandra gourd may have a small particle size.
  • the lignan content contained in the extract may be higher than when using Schisandra chinensis with large particle size.
  • the Dv10 value of Schisandra chinensis particles may be 20 ⁇ m to 30 ⁇ m, 22 ⁇ m to 28 ⁇ m, or 24 ⁇ m to 26 ⁇ m.
  • the Dv50 value of Schisandra chinensis particles may be 100 ⁇ m to 200 ⁇ m, 120 ⁇ m to 180 ⁇ m, 140 ⁇ m to 160 ⁇ m or 145 ⁇ m to 155 ⁇ m.
  • the Dv90 value of Schisandra chinensis particles may be 300 ⁇ m to 700 ⁇ m, 350 ⁇ m to 650 ⁇ m, 400 ⁇ m to 600 ⁇ m, or 450 ⁇ m to 550 ⁇ m.
  • Dv10 value refers to the average particle size at which 10% by volume of particles have a size smaller than that value.
  • Dv50 value refers to the average particle size at which 50% by volume of particles have a size smaller than that value.
  • Dv90 value refers to the average particle size at which 90% by volume of particles have a size smaller than that value.
  • the Dv10, Dv50 or Dv90 values may be values measured using a Malvern Mastersizer 3000 model micro particle size analyzer.
  • Solid-state fermentation is a fermentation that occurs when microorganisms grow on a wet substrate without running water.
  • GABA ⁇ -aminobutyric acid
  • lignan content may increase.
  • solid-state fermented Schisandra chinensis extract it may have superior DPPH radical scavenging ability, antibacterial activity, and AChE inhibitory ability compared to non-solid-state fermented Schisandra marigold extract.
  • Solid-state fermentation may be caused by at least one selected from the group consisting of Sanghwang mushrooms, Reishi mushrooms, shiitake mushrooms, Cordyceps sinensis, natto bacteria, and lactic acid bacteria.
  • solid-state fermented Schisandra gourd may be obtained by solid-state fermentation by inoculating Schisandra gourd with at least one selected from the group consisting of Sanghwang, Ganoderma, Shiitake, Cordyceps sinensis, Natto, and Lactobacillus.
  • the scientific name of Phellinus linteus is Phellinus linteus .
  • the scientific name of Reishi mushroom (common name) is Ganoderma sp .
  • the scientific name for shiitake mushrooms (common name) is Lentinus edodes .
  • the scientific name of Cordyceps sinensis (common name) is Cordyceps militaris .
  • the scientific name of Natto (common name) is Bacillus subtilis .
  • the scientific name for lactic acid bacteria is Lactobacillus plantarum .
  • the extract may be a substance separated by dissolving the substance to be extracted in a solvent, and specifically, an extract obtained by extraction treatment of the extraction subject, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a freeze-dried product of the extract, and the extract. It may include extracts of all formulations that can be formed using the extract itself and the extract, such as crude preparations, purified products, or mixtures thereof.
  • Extracts may differ in the extracted components or the content of the components depending on the type of solvent used for extraction, extraction method, etc.
  • the solid-state fermented Schisandra chinensis gourd extract may be extracted using a C1 to C4 lower alcohol as an extraction solvent.
  • the C1 to C4 lower alcohol may be an aqueous solution of C1 to C4 lower alcohol, which is a mixture of C1 to C4 lower alcohol and water.
  • the aqueous alcohol solution of C1 to C4 may be an aqueous solution of a lower alcohol of C1 to C4 of 40% to 80% by volume, 45% to 75% by volume, 50% to 70% by volume, or 55% to 65% by volume.
  • the C1 to C4 lower alcohol may be ethanol.
  • ethanol is used as an extraction solvent
  • the type or content of lignans extracted from solid-state fermented Schisandra chinensis peel may be greater than when extraction is performed using other solvents (e.g., distilled water).
  • the solid-state fermented Schisandra gourd extract may be extracted by ultrasonic extraction.
  • ultrasonic extraction is used as an extraction method, the type or content of lignans extracted from solid-state fermented Schisandra chinensis peel may be greater than when extraction is performed using other methods (e.g., heat extraction).
  • the solid-state fermented Schisandra chinensis gourd extract may be extracted by ultrasonic extraction using ethanol as an extraction solvent.
  • Fraction refers to the result obtained by performing fractionation to separate a specific component or specific group of components from a mixture containing various components.
  • the fraction of the solid-state fermented Schisandra chinensis extract may be a substance obtained by phase-separating the solid-state fermented Schisandra chinensis marigold extract with a solvent.
  • the solid-state fermented Schisandra chinensis marigold extract may be mixed with hexane, dichloromethane, or It may be a fraction fractionated with at least one solvent selected from the group consisting of ethyl acetate.
  • the present inventors obtained Schisandra chinensis gourd extract, which was solid-phase fermented with an aqueous ethanol solution, and sequentially fractionated it using hexane, dichloromethane, and ethyl acetate to produce a hexane-soluble fraction, a dichloromethane-soluble fraction, an ethyl acetate-soluble fraction, A water soluble fraction was prepared. Among the prepared fractions, it was confirmed that the hexane-soluble fraction contained a large amount of three types of lignans, which are active ingredients, and that the dichloromethane-soluble fraction and the ethyl acetate-soluble fraction contained large amounts of two types of lignans.
  • the fraction of the solid-state fermented Schisandra gourd extract may be a hexane fraction.
  • the fraction of the solid-state fermented Schisandra chinensis gourd extract may be a fraction obtained by first fractionating the extract with hexane and then secondarily fractionating it with dichloromethane.
  • the fraction of the solid-state fermented Schisandra chinensis gourd extract may be a fraction obtained by first fractionating the extract with hexane, second fractionation with dichloromethane, and third fractionation with ethyl acetate.
  • composition of the present invention has excellent antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement effects.
  • the antioxidant effect or anti-aging effect may be an effect obtained by the DPPH radical scavenging ability of the extract or fraction thereof of the present invention.
  • the antibacterial effect may be an effect obtained by the antibacterial activity of the extract or fraction thereof of the present invention against acne bacteria.
  • the anti-inflammatory effect may be obtained by the NO (nitric oxide) inhibitory ability of the extract or fraction thereof of the present invention.
  • the cognitive function improvement effect may be obtained by the AChE (acetylcholinesterase) inhibitory ability of the extract or fraction thereof of the present invention.
  • AChE acetylcholinesterase
  • improvement refers to any action that results in at least a reduction in the severity of the parameters associated with the condition being treated, such as symptoms.
  • composition of the present invention can be used for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement purposes.
  • composition of the present invention may be a pharmaceutical composition.
  • the pharmaceutical composition may be a pharmaceutical composition for preventing or treating acne, inflammatory diseases, insomnia, dementia, or Alzheimer's.
  • Acne may be caused by Propionibacterium acnes .
  • the inflammatory disease may be seborrheic dermatitis or a chronic inflammatory disease.
  • prophylaxis refers to prophylactic measures that result in any degree of reduction in the likelihood of developing the condition to be prevented or of reoccurrence or recurrence, including total prevention as well as slight, substantial or significant reduction in the likelihood of development or recurrence of the condition. Refers to an action, and the degree of probability reduction is at least a slight reduction.
  • treatment refers to treatment that results in a beneficial effect on the subject or patient suffering from the condition being treated, including cure as well as any degree of relief, including minor relief, substantial relief, major relief, the degree of relief being At least it is mild relief.
  • compositions may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, and sterile injectable solutions according to conventional methods, but are limited thereto. It doesn't work.
  • Carriers, excipients, and diluents that may be contained in the pharmaceutical composition include lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, and calcium phosphate. , calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto.
  • composition of the present invention may be a food composition.
  • Food compositions can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement.
  • Food compositions may contain common food additives, and their suitability as food additives is determined by the specifications and standards for the relevant item in accordance with the general provisions of the Food Additives Code and General Test Methods approved by the Food and Drug Administration, unless otherwise specified. It is decided by
  • Items listed in the Food Additives Code include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as dark pigment, licorice extract, crystalline cellulose, high-quality pigment, and guar gum; It includes, but is not limited to, mixed preparations such as L-glutamate sodium preparations, noodle added alkaline preparations, preservative preparations, and tar coloring preparations.
  • composition of the present invention may be a cosmetic composition.
  • the cosmetic composition of the present invention may contain fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, and foaming agents commonly used in the industry, depending on the quality or function of the final product. ), fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic activators, common in cosmetics. It may additionally contain auxiliaries commonly used in the fields of cosmetology or dermatology, such as any other ingredients used.
  • the cosmetic composition of the present invention includes soap, cleansing foam, cleansing cream, cleansing water, bath agent, skin, lotion, cream, essence, toner, astringent, emulsion, gel, lipstick, spray, shampoo, rinse, treatment, body cleanser, pack.
  • the present invention provides a method for producing a composition for antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement, comprising the step of extracting solid-state fermented Schisandra chinensis cucumber.
  • Schisandra gourd, solid-state fermentation, extraction, antioxidant, anti-aging, antibacterial, anti-inflammatory, cognitive function improvement and composition may be within the above-mentioned range.
  • the method for producing the composition includes the step of inoculating Schisandra chinensis with at least one selected from the group consisting of Sanghwang mushrooms, Reishi mushrooms, shiitake bacteria, Cordyceps sinensis, natto bacteria, and lactic acid bacteria into Schisandra cucumbers and subjecting them to solid-state fermentation to obtain the solid-state fermented Schisandra marsupials. It may further include ;. This is performed before the step of extracting the solid-state fermented Schisandra chinensis cucumber.
  • the method for producing the composition may further include fractionating the extracted Schisandra chinensis peel.
  • the fraction may be within the range described above.
  • composition prepared through the composition manufacturing method has excellent antioxidant, anti-aging, antibacterial, anti-inflammatory or cognitive function improvement effects.
  • Solid state fermentation is carried out by inoculating a liquid culture into a solid medium (Scisandra chinensis and Schisandra chinensis cucumber).
  • Solid-state fermentation requires a small amount of moisture, and mushroom fungi require lower water activity than bacteria or yeast, so mushroom fungi were selected as microorganisms used in solid-state fermentation.
  • the useful microorganisms Shangwang, Reishi, shiitake, Cordyceps, natto, lactic acid bacteria
  • Table 1 were inoculated onto standard media (PDA, PDB, MRS, NR media) and activated while culturing in a thermostat at 24°C. Strains that were expected to have fast mycelial growth and to contain a lot of effective substances after solid-state fermentation were selected.
  • Solid-state fermentation of Schisandra chinensis and Schisandra chinensis meal was performed by the following process. After drying Schisandra chinensis or Schisandra chinensis pumpkin and pulverizing it using a grinder, nutrient medium was added and mushroom bacteria (Shang Sangh, Ganoderma lucidum, Shiitake or Cordyceps sinensis), natto bacteria, or lactic acid bacteria were inoculated. When inoculated with mushroom bacteria, solid-state fermentation was performed for 6 weeks, and when inoculated with natto bacteria or lactic acid bacteria, it was performed for 5 weeks. The solid-state fermentation process of Schisandra chinensis and Schisandra chinensis meal is shown in Figure 3.
  • 100% DW thermal extract was prepared by the following processes 1 to 2.
  • 100% DW ultrasonic extract was prepared by the following processes 1 to 5.
  • the type and content of the active ingredient (lignan) in the Schisandra chinensis extract was confirmed after grinding according to the extraction solvent (60% EtOH or 100% DW) and extraction method (thermal extraction or ultrasonic extraction).
  • Table 4 below shows the results of component analysis of Schisandra chinensis extract after grinding according to various extraction solvents and extraction methods.
  • the type and content of the active ingredient (lignan) in Schisandra chinensis extract was confirmed after grinding according to the extraction solvent (60% EtOH or 100% DW) and extraction method (thermal extraction or ultrasonic extraction).
  • Table 5 shows the results of component analysis of Schisandra chinensis extract after grinding according to various extraction solvents and extraction methods, and is listed according to the grinding particle size of Schisandra chinensis bark.
  • the particle size of Schisandra chinensis porridge with the small particle size of 1) above and Schisandra chinensis porridge with the small particle size were measured.
  • the particle size was measured using a Micro Particle Size Analyzer (Mastersizer 3000, Malvern) under the following conditions: Dv10, Dv50, and Dv90 values:
  • Table 6 shows the particle size of Schisandra chinensis samples with small particle size in terms of Dv10, Dv50 or Dv90 values. A total of 5 measurements were made using a micro particle size analyzer, and the average value was recorded, which is expressed graphically in Figure 8.
  • Table 7 below shows the average values of the particle sizes 1 and 2 of FIG. 9 measured a total of 5 times using a ruler.
  • Table 8 below shows the particle size of 3 in FIG. 9 as Dv10, Dv50 or Dv90 values. A total of 5 measurements were made using a micro particle size analyzer, and the average value was recorded, which is expressed graphically in Figure 10.
  • Solid-state fermented Schisandra chinensis 100% DW thermal extract Solid-state fermentation microorganisms Schisandra chinensis situation Schisandra reishi Schisandra Shiitake Schisandra chinensis Cordyceps Schisandra Natto Schisandra lactic acid bacteria Average concentration (ppm) Schizandrin 287.4 ⁇ 10.0 300.5 ⁇ 10.8 304.3 ⁇ 11.8 300.6 ⁇ 8.0 284.4 ⁇ 8.8 288.9 ⁇ 18.1 Gomisin A 17.4 ⁇ 1.0 17.4 ⁇ 1.0 17.4 ⁇ 1.0 17.1 ⁇ 0.8 17.1 ⁇ 0.6 16.4 ⁇ 0.5 Gomisin N - - - - - - pH 2.79 2.76 2.77 2.79 2.80 2.80
  • the most lignans were extracted from Schisandra chinensis when solid-state fermented with Reishi, shiitake, and Cordyceps sinensis, and the most lignans were extracted from Schisandra chinensis when solid-state fermented with natto.
  • the most lignans were extracted from Schisandra chinensis when solid-state fermented with reishi, shiitake, natto, and lactic acid bacteria, and the most lignans were extracted from Schisandra chinensis when subjected to solid-state fermentation with natto. Heat extraction or ultrasonic extraction using 100% DW as an extraction solvent. When performed, only two types of lignans were detected, and it was found that there was no significant difference in content depending on the extraction method (Tables 9 to 12).
  • the mineral content of four types of solid-state fermented Schisandra chinensis 60% EtOH extracts and four types of solid-state fermented Schisandra chinensis 60% EtOH extracts was analyzed.
  • EA was performed to measure the content of organic matter
  • XRF was performed to measure the content of inorganic matter.
  • Table 15 below shows EA&XRF analysis data
  • Table 16 shows ICP-OES analysis data.
  • the mineral content through ICP-OES data shows that compared to the solid-state fermented Schisandra chinensis extract, the solid-state fermented Schisandra chinensis extract has increased P (phosphorus), S (sulfur), and Cu (copper) contents, as well as K (calcium), Ca (potassium), and The content of Mn (manganese) decreased.
  • the solid-state fermented Schisandra chinensis was subjected to ultrasonic extraction with 60% EtOH, and the egg extract was fractionated using non-polar solvents in the following order: Hexane, Dichloromethane, Ethyl Acetate, and Water (FIG. 14).
  • fractionation was performed by the following processes 1 to 4:
  • the standard product is measured starting from low concentration, ethanol is measured between the standard product and the standard product for washing, and then the sample is measured.
  • Cytotoxicity test antioxidant and anti-aging test, antibacterial test, anti-inflammatory test, and cognitive function improvement test were performed to analyze the efficacy of Schisandra chinensis 60% EtOH ultrasonic extract, which was solid-state fermented using Sanghwang, Ganoderma lucidum, Cordyceps sinensis, or natto.
  • the cell line used in the test was purchased from the Korea Cell Line Bank (KCLB).
  • the medium used was DMEM growth medium containing 10% Fetal Bovine Serum (FBS, Hyclone) and 5% Antibiotics/antimycotics (Sigma Aldrich).
  • FBS Fetal Bovine Serum
  • Antibiotics/antimycotics Sigma Aldrich.
  • Cell lines were cultured in an incubator controlled at 5% CO 2 , 37°C, and humidity. When the cells fill the culture dish to about 80%, wash with Pohsphate-buffered saline (PBS), add 1ml Trypsin/EDTA (Sigma Aldrich) to the dish, react at 37°C for 5 minutes, and dilute 10-fold with growth medium. After centrifugation, subculture was performed. The medium was changed every 48 hours to culture the cells. Table 24 below summarizes information on the cell lines used in the test.
  • RAW264.7 cells were suspended in DMEM growth medium, dispensed into a 96 well microplate at 3 ⁇ 10 4 cells/200 ⁇ l growth medium/well (i.e., 200 ⁇ l per well, repeated three times), and cultured in a CO2 incubator for 24 hours.
  • test solution After treatment with the test solution, it was cultured in a CO 2 incubator (37°C, humidified) for 24 hours.
  • the cytotoxicity test results of each extract are shown in Table 25 and Figure 18 below.
  • Table 25 the toxicity of each extract to RAW264.7 cells was shown through the IC50 value (mg/ml).
  • concentrations (ug/g) of the three lignans contained in each extract measured through the HPLC analysis method of 5-(4)-3) were described, and each lignan was determined based on the IC50 value of the extract and the measured lignan concentration. IC50 was calculated and reported.
  • DPPH 2,2-Diphenyl-1-picrylhydrazyl
  • Gallic Acid monohydrate Sigma Aldrich
  • 96 well type spectrophotometer SpectraMax ABS (Molecular Device)
  • ethanol distilled water, pipette, and e-tube were used.
  • Test sample preparation was carried out by the following process:
  • test sample extract
  • 60% ethanol a test sample solution
  • Gallic acid was used as a positive control for calculating equivalent values.
  • Gallic acid was prepared by dissolving it in 100% ethanol at a concentration of 1000 ⁇ g/mL and serially diluted in 60% ethanol.
  • the absorbance was measured without adding DPPH as a DPPH negative control, and then corrected for the absorbance value of the sample with DPPH added.
  • the inhibition rate was calculated using the following equation:
  • IC50 is calculated by drawing a curve using the inhibition rate (y) for each concentration of the sample (x), then substituting 50 for the inhibition rate (y) of the sample using the equation calculated from the trend line, and calculating the corresponding x value. did.
  • the antioxidant and anti-aging test results of each extract are shown in Table 26 and Figure 19 below.
  • Table 26 the DPPH radical scavenging ability of each extract was shown through IC50 value (mg/ml).
  • IC50 value of each lignan was calculated and reported based on the concentration (ug/g) of the three lignans contained in each extract and the IC50 value of the extract measured through the HPLC analysis method of 5-(4)-3). .
  • Schisandra chinensis root When looking at the antioxidant and anti-aging effects of Schisandra chinensis and Schisandra chinensis cucumbers that underwent solid-state fermentation, Schisandra chinensis root showed a lower IC50 value than Schisandra chinensis, except for the use of reishi, showing excellent free radical scavenging ability.
  • Petri dish (SPL cat.10090), Ethanol, Methyl paraben, 1 , a spectrophotometer was used.
  • test was conducted on a clean bench suitable for sterility testing in a separate space, and the instruments used for the test were maintained in a sterile state.
  • Preparation of medium Preparation of Brain Heart Infusion Agar: To measure growth inhibition of Propionibacterium acnes, add 18.5 g of Brain Heart Infusion (BHI), 500 mL of distilled water, and 9 g of agar (Agar, 1020-4405) and incubate at 121°C 15. After sterilizing for several minutes and cooling, the medium was solidified in a petri dish and used.
  • BHI Brain Heart Infusion
  • Agar Agar, 1020-4405
  • Bacterial smear 0.1 mL of Propionibacterium acnes solution (1x10 5 CFU/mL) was smeared on Brain Heart Infusion Agar.
  • the antibacterial efficacy was evaluated by converting it into the diameter of the inhibition ring (mm/mg) compared to the amount of treated sample.
  • the fermented one had a large reduction ring diameter value compared to the amount treated, showing excellent antibacterial efficacy against acne bacteria.
  • fermentation with Sanghwang, Cordyceps sinensis, and natto showed excellent antibacterial efficacy against acne bacteria.
  • Schisandra marigold showed a greater antibacterial ring diameter value compared to the amount treated than Schisandra chinensis, showing excellent antibacterial efficacy against acne bacteria.
  • the cell line used in the test was the same as in 6-(1)-1) above, and was cultured and used in the same manner.
  • the anti-inflammatory test was performed by the following procedure:
  • RAW264.7 cells were suspended in DMEM growth medium, dispensed into a 6-well plate at 1 ⁇ 10 6 cells/2 ml growth medium/well, and cultured in a CO 2 incubator (37°C, humidified) for 24 hours.
  • the culture medium was collected and passed through an 8,000 MWCO polysulfone centrifugal filter, and the filtrate was prepared as a sample.
  • a 96-well microplate was prepared, and according to the Griess reagent system (Promega) protocol, Nitrite standard solution was serially diluted two-fold in the concentration range between 100 and 0 uM and added to the wells, and the sample was also added (50 ul volume).
  • Anti-inflammatory efficacy was evaluated by calculating the x-value (nitrite concentration) after drawing up a nitrite standard curve, including the absorbance of the sample in the y-value of the calculated formula.
  • Schisandra chinensis natto 60% EtOH ultrasonic extract showed the highest anti-inflammatory efficacy, followed by Schisandra chinensis cordyceps 60% EtOH ultrasonic extract.
  • PBS Phosphate buffered saline
  • acetylcholinesterase human erythrocyte origin
  • DTNB 5,5 ⁇ -dithio-bis-2-nitrobenzoic acid
  • tacrine acetylcholine iodide
  • Test reagent preparation was carried out by the following process:
  • DTNB Dithiobisnitrobenzoic acid
  • Acetylthiocholine iodide was dissolved in PBS to reach 220mM.
  • AChE acetylcholinesterase
  • the cognitive function improvement test was performed by the following process:
  • AChE acetylcholinesterase
  • AChE inhibition rate (enzyme activity inhibition rate) was calculated using the following equation:
  • the cognitive function improvement test results of each extract are shown in Table 30 and Figure 23 below.
  • Table 30 the AChE inhibitory ability of each extract was shown through IC50 value (mg/ml).
  • IC50 value of each lignan was calculated and reported based on the concentration (ug/g) of the three lignans contained in each extract and the IC50 value of the extract measured through the HPLC analysis method of 5-(4)-3). .
  • the fermented one When looking at the efficacy of Schisandra chinensis in improving cognitive function according to the presence or absence of fermentation, the fermented one showed a low IC50 value, showing excellent cognitive function improvement efficacy. In particular, fermentation with Cordyceps sinensis and Sanghwang showed excellent efficacy in improving cognitive function.
  • Schisandra porridge When looking at the antibacterial efficacy of Schisandra chinensis and Schisandra chinensis porridge that underwent solid-state fermentation, Schisandra porridge showed a lower IC50 value than Schisandra chinensis, showing excellent efficacy in improving cognitive function.
  • Antibacterial test for acne bacteria evaluation expression 1 (Solid fermentation Schisandra chinensis - Schisandra chinensis)/ Schisandra chinensis * 100 2 (Solid-phase fermented Schisandra chinensis - Schisandra chinensis fruit)/ Schisandra chinensis fruit)*100 result 1 877.6% (Cordyceps sinensis solid fermented Schisandra chinensis - Schisandra chinensis after grinding)/ Schisandra chinensis after grinding *100 2 613.4% (Cordyceps sinensis solid fermented Schisandra chinensis - Schisandra chinensis)/ Schisandra chinensis*100
  • AChE inhibition activity test evaluation expression 1 (Solid fermentation Schisandra chinensis - Schisandra chinensis) / Schisandra fruit) ⁇ 100 2 (Solid fermentation Schisandra chinensis fruit - Schisandra chinensis fruit) / Schisandra chinensis) ⁇ 100 result 1 99.2% (Cordyceps sinensis solid fermented Schisandra chinensis - Schisandra chinensis after grinding)/ Schisandra chinensis after grinding)*100 2 93.5% (Cordyceps sinensis solid fermented Schisandra chinensis - Schisandra chinensis)/ Schisandra chinensis)*100
  • solid-state fermented Schisandra chinensis extract exhibits superior antioxidant, antibacterial, anti-inflammatory, and cognitive function improvement effects compared to Schisandra chinensis, crushed Schisandra chinensis, or Schisandra chinensis bark extract.

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Abstract

La présente invention concerne une composition pour améliorer la fonction antioxydante, anti-vieillissement, antibactérienne, anti-inflammatoire ou cognitive. Plus spécifiquement, la composition comprend un extrait de marc de Schizandra chinensis Baillon fermenté en phase solide ou une fraction de celui-ci et présente ainsi un excellent effet de piégeage de radicaux DPPH, un effet antibactérien contre acne bacillus, un effet inhibiteur de l'oxyde nitrique (NO) et un effet inhibiteur de l'acétylcholinestérase (AChE).
PCT/KR2023/002343 2022-03-29 2023-02-17 Composition d'amélioration de la fonction antioxydante, anti-vieillissement, antibactérienne, anti-inflammatoire ou cognitive contenant un extrait de marc de schizandra chinensis baillon fermenté en phase solide WO2023191310A1 (fr)

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