WO2021176707A1 - エラスターゼの阻害用経口組成物及びその利用、エラスターゼ阻害剤、エラスターゼ阻害剤を経口摂取するエラスターゼ活性阻害方法 - Google Patents

エラスターゼの阻害用経口組成物及びその利用、エラスターゼ阻害剤、エラスターゼ阻害剤を経口摂取するエラスターゼ活性阻害方法 Download PDF

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Publication number
WO2021176707A1
WO2021176707A1 PCT/JP2020/009785 JP2020009785W WO2021176707A1 WO 2021176707 A1 WO2021176707 A1 WO 2021176707A1 JP 2020009785 W JP2020009785 W JP 2020009785W WO 2021176707 A1 WO2021176707 A1 WO 2021176707A1
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Prior art keywords
elastase
okra
inhibiting
seeds
skin
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PCT/JP2020/009785
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English (en)
French (fr)
Japanese (ja)
Inventor
郁郎 田中
昌代 木村
勉 野崎
石原　健夫
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BHN Co Ltd
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BHN Co Ltd
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Priority to EP20923599.3A priority Critical patent/EP4118978A4/en
Priority to CN202080098184.3A priority patent/CN115243563B/zh
Priority to PCT/JP2020/009785 priority patent/WO2021176707A1/ja
Priority to JP2022504931A priority patent/JP7513299B2/ja
Publication of WO2021176707A1 publication Critical patent/WO2021176707A1/ja
Anticipated expiration legal-status Critical
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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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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)
    • A61K36/577—Malvaceae (Mallow 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
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
    • 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
    • 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
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/20—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof consisting of whole seeds or seed fragments
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/30—Mashed or comminuted products, e.g. pulp, pastes, meal, powders; Products made therefrom, e.g. blocks, flakes, snacks; Liquid or semi-liquid products

Definitions

  • the present invention relates to an oral composition for inhibiting elastase and its use, which comprises a processed product of okra (Abelmoschus esculentus) seeds, an elastase inhibitor, and a method for inhibiting elastase activity by orally ingesting an elastase inhibitor. ..
  • the present invention relates to an oral composition for inhibiting elastase containing the cyclic peptide "OF4949-II" as an active ingredient and its use, an elastase inhibitor, and a method for inhibiting elastase activity by orally ingesting an elastase inhibitor.
  • Connective tissue is also called supportive tissue, and in a broad sense, bone, cartilage, blood, fat, etc. may also be classified as connective tissue.
  • epithelial tissues and supporting tissues have been researched and developed for a long time in skin, bones, fats, etc.
  • skin tension and elasticity have decreased (wrinkles, as part of skin aging symptoms).
  • the mechanism of occurrence of sagging, etc.) is being elucidated, and it is said that elastin (elastic fibers) is greatly involved in wrinkles and sagging of the skin.
  • Elastin is a type of fibrous high molecular protein existing in connective tissue.
  • Collagen which is a high molecular protein, is bound to each other in a lattice pattern, and mucopolysaccharides such as hyaluronic acid and chondroitin sulfate are contained in the gaps.
  • mucopolysaccharides such as hyaluronic acid and chondroitin sulfate are contained in the gaps.
  • This extracellular matrix structure is derived from external factors such as support of cells and skin tissue, water retention in intercellular spaces, retention of skin lubricity and flexibility, ultraviolet rays, dry environment, mechanical irritation and damage, and microbial infection. It plays a role of protecting the skin. It is known that these proteins and extracellular components are produced by fibroblasts in vivo (Non-Patent Document 1).
  • the elastin content in living tissues is generally recognized to be about 78 to 80% in the term ligament, about 50% in the artery, about 20% in the lung, and about 2 to 5% in the dermis. Also, as one of the characteristics of elastin, unlike collagen and hyaluronic acid, the amount increases with growth at almost 0% at birth of humans, and peaks in the mid-20s. After that, it started to decrease, and it is said that it will be less than 50% after the 40s. This suggests that elastin in living tissues may be decomposed or denatured by metabolic changes associated with aging, ultraviolet rays, and other factors, and one of the causes is increased activity or excess of elastase, an enzyme that decomposes elastin. Guidance etc. is inferred.
  • Patent Document 1 an aqueous component obtained from okra seeds is used as an active ingredient that promotes the proliferation of fibroblasts.
  • Patent Document 2 various studies have been conducted to prevent the reduction of the content of elastin, collagen, etc. in the dermis, and various reports and proposals have been made.
  • the retinol glycoside is disclosed as (Patent Document 2).
  • side effects due to overdose are also known for retinol derivatives, which is not sufficient in terms of safety.
  • Patent Document 3 extract of broomrape
  • Patent Document 5 resveratrols extracted from grape buds and vines
  • Patent Document 6 extraction of Mannentake
  • Patent Document 8 soybean protein hydrolyzate
  • Okra Abelmoschus manicure
  • Okra is a plant belonging to the genus Okras of the Malvaceae family, and has a long history of growing in tropical to temperate regions of the world, cultivated as a vegetable, and used for food.
  • fruits (pods) containing immature white to yellowish white seeds are eaten as fresh vegetables.
  • Patent Document 1 As an attempt to industrially utilize the extract and ingredients obtained by processing okra, Patent Document 1, the hyaluronic acid synthesis promoter containing the okra extract, and cosmetics and foods and drinks containing the agent (Patent Document 9). ), An antiaging skin external preparation containing an oligopeptide derived from okra seeds and a specific plant extract (Patent Document 10) and the like have been proposed. However, no mention is made of the association between okra seeds and processed products and elastin and elastase.
  • JP-A-2018-115143 JP-A-10-158290 (Claim 1 et al.) Japanese Unexamined Patent Publication No. 2012-201615, Japanese Unexamined Patent Publication No. 2009-263279 Japanese Unexamined Patent Publication No. 2008-239576 Japanese Unexamined Patent Publication No. 2005-53873 Japanese Unexamined Patent Publication No. 2005-23021 Japanese Unexamined Patent Publication No. 2004-182687 Japanese Unexamined Patent Publication No. 2004-51533 Japanese Unexamined Patent Publication No. 2008-74757 Japanese Unexamined Patent Publication No. 02-288890
  • the present invention recovers the reduction of elastin in living tissues caused by external factors such as ultraviolet rays and active oxygen, and deterioration of metabolic function of the living body, and improves skin aging symptoms and the like. It is an object of the present invention to provide an oral composition for inhibiting elastase and its use, an elastase inhibitor, and a method for inhibiting elastase activity by orally ingesting an elastase inhibitor.
  • the present inventors have conducted diligent studies on a material that inhibits the activity of fibroblast-derived elastase and a processing method thereof, and as a result, found that a processed product of okra seeds is effective. I found it. Furthermore, as a result of continuing research and development on a component that inhibits the activity of fibroblast-derived elastase and a method for purifying it, the present inventors used the cyclic peptide "OF4949-II" obtained from ripe okra seeds as an active ingredient. We found that there was something, and came to complete the present invention.
  • a composition for inhibiting elastase which comprises a processed product of okra (Abelmoschus esculentus) seeds.
  • the seed is a ripe seed
  • the processed product is preferably any of an extract of okla seed, the extraction residue, an enzymatic decomposition product or a sprout, and the treatment of the extraction is hot water extraction.
  • the enzymatic degradation treatment is hydrolysis using an alkaline protease and / or a neutral protease
  • the elastase is preferably derived from skin fibroblasts.
  • the composition is orally ingested or administered.
  • a composition for inhibiting elastase which comprises a specific compound derived from okra seeds as an active ingredient.
  • the seeds are ripe seeds, and that the elastase is derived from skin fibroblasts.
  • the specific compound is preferably a cyclic peptide "OF4949-II", and the peptide is preferably represented by the following formula I.
  • composition characterized by inhibiting the activity of elastase by ingesting or administering the composition.
  • the composition is orally ingested or administered and used, and it is desirable that the oral composition is in the form of a food or drink. Further, it is desirable that the composition is used for increasing elastin based on the elastase activity inhibitory action of the skin tissue, and further, it is desirable that the composition is used for improving the aging symptoms of the skin tissue.
  • the composition containing the peptide is orally ingested or administered and used, and it is desirable that the oral composition containing the peptide is in the form of food and drink. Further, it is desirable that the composition is used for increasing elastin based on the elastase activity inhibitory action of the skin tissue, and further, it is desirable that the composition is used for improving the aging symptoms of the skin tissue.
  • An elastase inhibitor characterized by blending the composition for inhibiting elastase.
  • a method for inhibiting elastase activity which comprises orally ingesting the above-mentioned elastase inhibitor.
  • the hot water extract, its residue, protein hydrolyzate and sprout, which are processed products obtained from ripe okra seeds according to the present invention, are effective for the cyclic peptide "OF4949-II" which is a specific compound derived from okra seeds.
  • the composition for inhibiting elastase of the present invention and the elastase inhibitor containing the peptide promote skin turnover to improve skin troubles and aging symptoms (wrinkles, stains, dullness, freckles, sagging, etc.). It is possible to contribute to the promotion of beautiful skin.
  • the composition for inhibiting elastase of the present invention and the inhibitor obtained by blending the composition are, in particular, in the fields of foods and drinks, pharmaceuticals, animal feeds, etc. It can be effectively used for skin improvement in the form compounded in the product.
  • the composition for inhibiting elastase of the present invention has an action of inhibiting elastase, which is an enzyme that decomposes elastin present in living tissues, especially in the dermis tissue of the skin, and contains a processed product of okra seeds. It is characterized by doing.
  • the composition for inhibiting elastase of the present invention contains the cyclic peptide "OF4949-II", which is a specific compound derived from okra seeds, as an active ingredient.
  • Okra generally has varieties with pod shapes such as pentagon, octagon, and round, and varieties with colors such as green, purple-red, and white.
  • the okra according to the present invention is not limited to these types, and any okra can be used.
  • Ripe seeds refer to seeds harvested from the grown fruits (pods) of plants or seeds that spontaneously scatter, and are dark green to dark brown in size with a diameter of about 5 mm and germinate when sown. Has the ability to do.
  • immature seeds are contained in growing pods and have a diameter of about 2 to 4 mm in white to yellowish white, and are usually used together with pods for food.
  • Ripe okra seeds are commercially available from seed companies for growing okra, and it is convenient to use them.
  • okra seeds as a processed product described below, that is, an extract of okra seeds, the extraction residue, an enzyme hydrolase or a sprout.
  • the okra seed extract can be produced, for example, by the following processing.
  • the dehulled or non-dehulled seeds are coarsely pulverized or finely pulverized, water is added to obtain a dispersion liquid, and then the extraction treatment is carried out at room temperature or under heating while allowing the seeds to stand or stirring appropriately.
  • the extract is treated by a conventional method such as centrifugation or filtration to remove insoluble matter, and more preferably, it is concentrated and dried or freeze-dried at a low temperature of room temperature or lower to relate to the present invention. Extracts can be produced.
  • the aqueous dispersion when producing a hot water extract, is set at about 30 to about 90 ° C., more preferably about 40 to about 60 ° C., and for about 10 minutes to several days.
  • the temperature is set to about 10 to about 30 ° C., more preferably about 15 to about 25 ° C., and more preferably about 30 minutes to several days. May be soaked for several hours to one day for extraction treatment.
  • the extraction residue of okra seeds can be produced by concentrated drying or freeze-drying the insoluble matter removed in the extraction step by a conventional method.
  • the degree of drying is preferably such that the water content is 5% by mass or less in order to prevent contamination due to the generation of mold and the like. It is desirable that the residue of the present invention is a residue (low temperature extraction residue) separated when the above-mentioned low-temperature extract is produced in relation to the desired effect of the present invention.
  • the enzymatically decomposed okra seed can be produced by processing as follows. That is, the okra seeds were crushed into an aqueous dispersion, and about 0.1 to about 5% by mass, more preferably about 0.5 to about 2% by mass of protein hydrolase was added to the aqueous dispersion. Hydrolyze for about 10 minutes to several days, more preferably for 30 minutes to several hours, at room temperature or warming (about 30 to about 90 ° C., more preferably about 40 to about 60 ° C.), standing or stirring appropriately. Have the decomposition process performed. After the treatment, the enzyme is heat-inactivated by a conventional method, then centrifuged or filtered to separate insoluble matter, and preferably concentrated, dried or freeze-dried at room temperature or lower. Can be done. In the present invention, instead of the aqueous dispersion, the okra seed extract or extraction residue obtained by the treatment as described above is dissolved or dispersed in water and subjected to the same enzymatic treatment. May be good.
  • the protein hydrolase can be of any kind or type, but in the present invention, alkaline protease and / or neutral protease is preferable, and alkaline protease is even more desirable.
  • the alkaline protease can be used as long as it has an action at a substrate pH of about 6.5 to 12, and the neutral protease has an action at a substrate pH of about 5 to 8. If there is, there is no problem.
  • each protease may be used not only by one type but also by a combination of two or more types, and it is convenient to use a commercially available product as described later.
  • alkaline proteases The following specific examples can be given as alkaline proteases. However, the present invention is not limited thereto. "Protin SD-AY10", “Protease P” Amano “3SD”, “Proleather (registered trademark) FG-F” (all manufactured by Amano Enzyme Co., Ltd.), “Alcalase (registered trademark) 2.4LFG” (Novozymes) "Orientase (registered trademark) 22BF” (manufactured by HBI Co., Ltd.), “Aroase (registered trademark) XA-10” (manufactured by Yakult Pharmaceutical Co., Ltd.), “Sumiteam (registered trademark) MP” ( Shin Nihon Kagaku Kogyo Co., Ltd.), “Bioprotease OP, SP-20FG, AL-15FG, 30G, APL-30 and 30L” (manufactured by Nagase ChemteX Corporation), “OPTIMASE (registered trademark)
  • Okra seed sprout is obtained by germinating seeds by a known method, growing them in a radish sprout shape, cultivating green leaves until the stem height is about 5 to 15 cm, and then washing with water and then drying or freeze-drying.
  • the processed product obtained by the treatment as described above is a protein contained in okla seeds, a polysaccharide protein complex, a polysaccharide and / or peptides, amino acids, oligosaccharides, monosaccharides or oligos obtained by hydrolyzing these. It is an extremely complex composition containing various components such as a conjugate of a saccharide and a peptide or an amino acid.
  • the cyclic peptide "OF4949-II” is contained as an active ingredient. It is known that the cyclic peptide "OF4949-II” strongly inhibits the activity of aminopeptidase B and is produced by a microorganism belonging to the genus Penicillium (Patent Document 11), but in ripe seeds of Penicillium. There are no reports that it is included.
  • the cyclic peptide "OF4949-II" can be produced, for example, by the following purification treatment.
  • the ripe okra seeds are extracted while being heated, allowed to stand, or stirred appropriately.
  • this extract is treated by a conventional method such as centrifugation or filtration to remove insoluble matter, and more preferably, it is concentrated and dried or freeze-dried at a low temperature of room temperature or lower to obtain an extract.
  • the obtained extract is concentrated by flash chromatography or high performance liquid chromatography to concentrate the desired fraction, and purification is repeated.
  • the above-mentioned processed okra seed product is contained as it is, or in combination with known excipients such as dextrin, cellulosic, gelatin, purified water, etc., to contain the composition for inhibiting elastase of the present invention.
  • known excipients such as dextrin, cellulosic, gelatin, purified water, etc.
  • Liquid, powder, granular and capsule inhibitors can be prepared.
  • the content of the processed okra seed product in this agent is approximately 20% by mass or more, and if it is less than this, it becomes difficult to exhibit the desired effect of the present invention. Therefore, the processed product / excipient (mass ratio) of the okra seed is used. It is preferably 100 to about 20/0 to about 80, and even more preferably 100 to about 50/0 to about 50.
  • composition for inhibiting elastase of the present invention and the elastase inhibitor containing the same can suppress the action of elastase in the skin tissue and prevent the reduction of elastin by ingesting or administering the composition of the skin. It can be used as a beauty means for improving the trouble, recovering from aging symptoms, and promoting beautiful skin.
  • foods and drinks, pharmaceuticals, animal feeds and the like obtained by molding the elastase inhibitor of the present invention into a solid form (powder or granule), a gel form (jelly), a paste form or a liquid form (beverage or drink), etc. It is possible to make it a product of.
  • known additives surfactants, thickeners, antioxidants, colorants, fragrances, etc.
  • fibrous buds fibrous buds
  • Various products are prepared by a conventional method by appropriately using a material derived from animals and plants known to have a cell growth promoting action, an extracellular component production promoting action such as collagen and hyaluronic acid, a skin aging preventing action, and a skin beautifying promoting action). It is possible.
  • Comparative sample 1 As the comparative sample 1, a commercially available product derived from lychee seed (manufactured by Oriza Yuka Co., Ltd., trade name: "lychee seed extract-WSP”), which is a material known to have an elastase inhibitory effect, was used.
  • lychee seed extract-WSP a commercially available product derived from lychee seed
  • Comparative sample 2 As Comparative Sample 2, a commercially available product of resveratrol-containing composition (manufactured by BHN Co., Ltd., trade name: “resveratrol ⁇ ”), which is a material known to have an elastase-inhibiting effect, was used.
  • resveratrol ⁇ a commercially available product of resveratrol-containing composition
  • Comparative sample 3 As Comparative Sample 3, a commercially available product of Button Siler leaf powder, which is a known plant-derived material (manufactured by BHN Co., Ltd., trade name: “Button Siler leaf powder”) was used.
  • Comparative sample 4 As the comparative sample 4, a commercially available product of sesame leaf powder (manufactured by BHN Co., Ltd., trade name: “sesame leaf powder”), which is a known plant-derived material, was used.
  • Comparative sample 5 As the comparative sample 5, a commercially available product of okra seed powder (seller: Matsumoto Kosho Co., Ltd., trade name: "Myoxinol LS 9736”), which is a known material derived from okra, was used.
  • Production example 5 The same ripe seeds as in Production Example 1 were soaked in water at 0 ° C. to room temperature for 8 to 20 hours to remove water, then sown in a cultivation container and stored in a dark room at room temperature to 30 ° C. and humidity of 75 to 90%. It was germinated by spraying water as appropriate. After germination, the plants were transferred to a house and sprinkled with water at room temperature of 25 to 30 ° C. in sunlight to obtain radish sprouts 5 to 7 days later. This was washed with water, freeze-dried and crushed to produce a crushed sprout product (Sample 5).
  • Comparative manufacturing example 1 In Production Example 1, an enzymatic decomposition product (comparative sample 6) was obtained by the same treatment except that the ripe seeds were replaced with immature seeds.
  • Comparative manufacturing example 2 Same as in Production Example 1 except that the ripe seeds were replaced with a commercially available product (manufactured by Yale Co., Ltd., trade name: "Okra powder”) derived from the edible portion of okra (pods containing immature seeds; the same applies hereinafter). To obtain an enzymatic decomposition product (comparative sample 7).
  • a commercially available product manufactured by Yale Co., Ltd., trade name: "Okra powder” derived from the edible portion of okra (pods containing immature seeds; the same applies hereinafter).
  • Comparative manufacturing example 3 In Production Example 2, the extract (comparative sample 8) was treated in the same manner except that the ripe seeds were replaced with a commercially available product derived from the edible part of okra (manufactured by Yale Co., Ltd., trade name: "Okra powder”). Obtained.
  • Test Example 1 The effect of each of the above samples on elastase activity was investigated by the method described below.
  • Test Example 1 Elastase inhibitory action (1) The above-mentioned sample 1 and comparative samples 1 to 3 were used as test samples for measuring the elastase activity. Each test sample was dissolved in 100 mM Tris-hydrochloric acid buffer (pH 8.0) to prepare a test solution having a final concentration of 40.0 to 1000.0 ⁇ g / ml. At this time, if it was difficult to dissolve, DMSO was added within 10%.
  • Test Example 2 Elastase inhibitory action (Part 2)
  • the effects of processed products other than enzymatically decomposed okra seeds and processed products other than ripe seeds on elastase inhibitory action were investigated.
  • the measurement of the elastase inhibitory activity and the evaluation of the degree of the inhibitory effect were carried out by the same method as in Test Example 1, but the final concentration of the test solution was set to 0.4 to 10.0 ⁇ g / ml.
  • the above-mentioned sample 1 samples 2 to 5, and comparative samples 5 to 8 were used.
  • Test Example 3 Human Monitor Test Forty adult female volunteers (30-55 years old, average age: 44.5 years old) who consented to the following tests were asked to be divided into 20 people per group, 1 One group received sample 1 (100 mg) and the other group took placebo (dextrin 100 mg) orally twice a day, which was continued for 4 weeks. Regarding changes in the skin before and after ingestion, use items such as dry skin, stains, wrinkles, swelling, freezing, sagging, nasolabial folds, makeup glue, and pore opening, and conduct an questionnaire survey to see if there is any improvement. evaluated.
  • the skin improved compared to before ingestion 17 persons, did not change: 2 persons, deteriorated 1 person, and in the placebo group, the skin improved: 2 persons, did not change. : 14 people, 4 people who got worse.
  • the 17 people with altered skin had two or more choices for improvement, especially for wrinkles, swelling, sagging, and nasolabial folds.
  • improvement of dry skin and pore opening which was not seen in the placebo group, was also selected.
  • the results suggest that the processed okra ripe seeds according to the present invention are useful for improving the skin by inhibiting the activity of elastase.
  • Test Example 1 The effect of each of the above fractions on elastase activity was investigated by the method described below.
  • Test example Elastase inhibitory action
  • Each of the above fractions was used as a test sample for measuring elastase activity.
  • the test method was carried out in the same manner as in Test Example 2 above, with the final concentration of the test solution set to 0.4 to 10.0 ⁇ g / ml.
  • the fraction F was treated and purified as follows.
  • the fraction F was concentrated and dried, and dissolved in distilled water to a concentration of about 1 to 5 mg / mL.
  • gradient analysis was performed using a C18 column 5C 18- MS-II (Nacalai Tesque) at a flow rate of 0.7 mL / min using high performance liquid chromatography (HPLC), and 5 types of fractions (in order of elution (in order of elution)).
  • HPLC high performance liquid chromatography
  • FIG. 4 shows a chromatogram of the purified product when fraction (5) -1 is fractionated at a concentration of about 10 mg / mL.
  • the vertical axis represents the ultraviolet absorption intensity (mV)
  • the horizontal axis represents the holding time (min).
  • the purified product obtained by the above treatment was subjected to direct introduction / mass spectrometry (DI / MS) and nuclear magnetic resonance analysis (NMR) to estimate a molecular weight of 472 and a molecular formula of C 22 H 24 N 4 O 8. It was confirmed that the cyclic peptide represented by the formula I was "OF4949-II" (CAS number: 93375-50-9).
  • Prototype example 1 Hard capsule preparation The above sample 1 was used in a capsule filling machine to make a trial production of a gelatin-coated hard capsule preparation having a content of 100 mg per capsule by a conventional method. When the hard capsule preparation (containing the ripe okra seeds of Production Example 1) was orally ingested in a monitor test, it was found to be useful for improving the skin. Therefore, this tablet can be used as an orally ingestible dietary supplement, medicine or animal feed for skin improvement.
  • Prototype Example 2 Tablets A tablet was prototyped by tableting the following raw materials by a conventional method. Here, as the ripe okra seeds, any one of the above samples 1 to 5 was used. All of these tablets are stable and easy to take, and can be used as dietary supplements and pharmaceuticals. (Mixed raw material) (Mass per tablet (mg)) 1. 1. Okra ripe seeds 100 2. Dextrin 100 3. 3. Valley starch 49 4. Microcrystal Cellulose 20 5. Synthetic aluminum silicate 30 6. Calcium stearate 1
  • Prototype Example 3 Dietary supplement The sample 1 was 100 mg, leucine was 400 mg, isoleucine was 250 mg, valine was 200 mg, and arginine was 300 mg. This dietary supplement can be used not only for humans but also for pet foods.
  • Prototype example 4 Food (noodles) The sample 1 was 10 g, buckwheat flour was 350 g, and strong flour was 150 g. While mixing these, they were sieved and 150 ml of water was mixed to make a buckwheat prototype. Compared with the commercial product, this had no discomfort in flavor and texture.
  • Prototype Example 5 Soft drinks and drinks 500 mg or 100 mg of the sample 1 or 2 was added to 100 mL of commercially available soft drinks and drinks and mixed sufficiently to make a prototype drink. This beverage had no color in flavor and storage stability compared to the original soft drinks and drinks. It can be used as a soft drink or drink.
  • the processed product obtained from the ripe seeds of okra in particular, the cyclic peptide "OF4949-II" obtained from the ripe seeds of okra has an action of inhibiting the elastase activity derived from fibroblasts, and therefore it is taken orally.
  • the original physiological function of the skin is restored, and it can be effectively used as foods and drinks, pharmaceuticals, animal feeds, etc., which are useful for improving skin troubles, promoting beautiful skin, and early recovery of skin damage.

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PCT/JP2020/009785 2020-03-06 2020-03-06 エラスターゼの阻害用経口組成物及びその利用、エラスターゼ阻害剤、エラスターゼ阻害剤を経口摂取するエラスターゼ活性阻害方法 Ceased WO2021176707A1 (ja)

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EP20923599.3A EP4118978A4 (en) 2020-03-06 2020-03-06 ORAL COMPOSITION FOR INHIBITING ELASTASE AND USE THEREOF, ELASTASE INHIBITOR AND METHOD FOR INHIBITING ELASTASE ACTIVITY BY ORAL INTAKE OF AN ELASTASE INHIBITOR
CN202080098184.3A CN115243563B (zh) 2020-03-06 2020-03-06 用于抑制弹性蛋白酶的口服组合物及其应用、弹性蛋白酶抑制剂、口服摄入弹性蛋白酶抑制剂的抑制弹性蛋白酶活性的方法
PCT/JP2020/009785 WO2021176707A1 (ja) 2020-03-06 2020-03-06 エラスターゼの阻害用経口組成物及びその利用、エラスターゼ阻害剤、エラスターゼ阻害剤を経口摂取するエラスターゼ活性阻害方法
JP2022504931A JP7513299B2 (ja) 2020-03-06 2020-03-06 エラスターゼの阻害用経口組成物及びその利用、エラスターゼ阻害剤、エラスターゼ阻害剤を経口摂取するエラスターゼ活性阻害方法

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CN116712361A (zh) * 2023-06-14 2023-09-08 遂宁市泽汐生物科技有限公司 列当提取物在制备改善皮肤弹性和皱纹组合物中的应用

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CN115243563B (zh) 2025-03-25
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