WO2018066707A1 - Inhibiteur de vieillissement, inhibiteur de calcification de tissu mou et inhibiteur de destruction de tissu pulmonaire - Google Patents

Inhibiteur de vieillissement, inhibiteur de calcification de tissu mou et inhibiteur de destruction de tissu pulmonaire Download PDF

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WO2018066707A1
WO2018066707A1 PCT/JP2017/036547 JP2017036547W WO2018066707A1 WO 2018066707 A1 WO2018066707 A1 WO 2018066707A1 JP 2017036547 W JP2017036547 W JP 2017036547W WO 2018066707 A1 WO2018066707 A1 WO 2018066707A1
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inhibitor
aging
isothiocyanate
calcification
mice
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PCT/JP2017/036547
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English (en)
Japanese (ja)
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陽一 鍋島
千秋 安部
義浩 宇都
平松　隆司
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株式会社 Ｐａｌ
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Priority to JP2018543997A priority Critical patent/JP7068704B2/ja
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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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/26—Cyanate or isocyanate esters; Thiocyanate or isothiocyanate esters
    • 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)
    • A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
    • 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
    • A61K8/46—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing sulfur
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • 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
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • 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
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • 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
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • 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

Definitions

  • the present invention relates to an aging inhibitor containing ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof, a soft tissue calcification inhibitor, a lung tissue destruction inhibitor, a food or drink containing them, a pharmaceutical product, and
  • the present invention relates to cosmetics and methods for using them.
  • 6-methylsulfinyl hexyl isothiocyanate (hereinafter also referred to as 6-MSITC) is one of the perfumes contained in wasabi and has various anti-inflammatory and anti-inflammatory pharmacological actions so far. It is known (Non-Patent Documents 3 and 4).
  • Patent Document 1 6-methylsulfinylhexyl isothiocyanate activates NAD (P) H: quinone oxidoreductase (NQO1) to adjust the NAD (P) + / NAD (P) H ratio, It has been suggested that it can be used for various diseases that can arise from excessive energy (Patent Document 1: JP 2009-526839).
  • Patent Document 2 describes that 6-methylsulfinylhexyl isothiocyanate suppresses the production of TNF- ⁇ , and is suggested to be used for inflammatory diseases such as osteoarthritis and rheumatoid arthritis. (Patent Document 2: JP 2009-132635 A).
  • Patent Document 3 6-methylsulfinylhexyl isothiocyanate suppresses increase in blood corticotropin (ACTH) concentration in the blood after stress and / or physical stress, and increases in cytokines and chemokines. has been described, and is suggested to be used for diseases caused by stress (Patent Document 3: JP-A-2009-126826).
  • ACTH blood corticotropin
  • ⁇ -methylsulfinylalkylisothiocyanate has an aging inhibitory action, a soft tissue calcification inhibitory action, and a lung tissue destruction inhibitory action.
  • the problem to be solved by the present invention is to provide a novel aging inhibitor, a soft tissue calcification inhibitor, and a lung tissue destruction inhibitor, foods and drinks, pharmaceuticals and cosmetics containing the same, and methods for using them. That is.
  • ⁇ -methylsulfinylalkylisothiocyanate has an aging inhibitory action, a soft tissue calcification inhibitory action, and a lung tissue destruction inhibitory action
  • the present invention has been completed.
  • the present inventors have also found that ⁇ -methylsulfinylalkylisothiocyanate has a calpain-1 activation inhibitory action, which brings about an aging inhibitory action, a soft tissue calcification inhibitory action, and a lung tissue destruction inhibitory action. I found out.
  • the aging inhibitor of the present invention contains ⁇ -methylsulfinylalkyl isothiocyanate or a physiologically acceptable salt thereof.
  • the mineralization inhibitor of the present invention contains ⁇ -methylsulfinylalkyl isothiocyanate or a physiologically acceptable salt thereof.
  • the lung tissue destruction inhibitor of the present invention contains ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof.
  • the present invention can provide an aging inhibitor, a soft tissue calcification inhibitor, and a lung tissue destruction inhibitor containing ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof. Taking this as a food or drink or as a cosmetic, for example, improving a state related to aging, suppressing calcification of soft tissue, or suppressing destruction of lung tissue be able to.
  • 6 shows the aging inhibitory effect of 6-MSITC on aging model mice ( ⁇ -klotho KO mice).
  • 6 shows the effect of wasabi-containing 6-MISTC on emphysema in Wild type mice and ⁇ -klotho KO mice.
  • 6 shows the ectopic calcification-inhibiting effect of wasabi-containing 6-MSITC in Wild type mice and ⁇ -klotho KO mice.
  • 6 shows the activation inhibitory effect of Calpain-1 in an aging model mouse ( ⁇ -klotho KO mouse) of 6-MSITC.
  • 6 shows the effect of suppressing Calpain-1 activation in 6-MSITC Wild type mice and ⁇ -klotho KO mice.
  • 6-MSITC shows the aging inhibitory effect of 6-MSITC on aging model mice (SAMP1 mice) (beam balance test).
  • 6-MSITC shows an aging inhibitory effect on aging model mice (SAMP1 mice) (external appearance of skin).
  • 6 shows aging inhibitory effect of 6-MSITC on aging model mice (SAMP1 mice) (histological analysis).
  • 6 shows the effect of suppressing activation of Calpain-1 by 6-MSITC.
  • 6 shows the effect of suppressing the degradation of Calpastatin by 6-MSITC.
  • ⁇ -methylsulfinylalkyl isothiocyanate may be a chemically synthesized substance or a natural product as an extract obtained from a cruciferous plant.
  • Specific examples of these substances include 5-methylsulfinylpentyl isothiocyanate, 6-methylsulfinyl hexyl isothiocyanate, 7-methylsulfinyl heptyl isothiocyanate, and 8-methylsulfinyl octyl isothiocyanate.
  • 6-methylsulfinylhexyl isothiocyanate is preferable.
  • ⁇ -methylsulfinylalkyl isothiocyanate is a natural product such as wasabi, horseradish, cabbage, watercress, brussels sprouts, cauliflower, radish, leach radish, rapeseed, broccoli, Takana, mustard, turnip, Chinese cabbage, etc.
  • this wasabi Wisabia Japana
  • 6-methylsulfinylhexyl isothiocyanate is more preferable, and either this wasabi leaf or this wasabi rhizome may be used. Particularly preferred.
  • an amino group is introduced by using the Gabriel method to produce N- ( ⁇ -methylthioalkyl) -phthalimide, and hydrazine hydrate is added to the resultant to reflux to obtain ⁇ -methylthioalkylamine.
  • ⁇ -methylthioalkyl isothiocyanate obtained via thiuram disulfide was oxidized with mCPBA to oxidize the methylthio group, and ⁇ Obtain methylsulfinylalkylisothiocyanate.
  • the plant body when extracting from a Brassicaceae plant containing ⁇ -methylsulfinylalkylisothiocyanate, the plant body is subjected to extraction pretreatment by physical means such as grinding or grated water, methanol, ethanol, acetone, ethyl acetate. Extraction with an organic solvent such as diethyl ether, dichloromethane or dichloroethane, or extraction with a distillation method such as steam distillation or molecular distillation is preferred, but it is not particularly limited to these methods.
  • the specific extraction method of this wasabi with an organic solvent is as follows. After the rhizome of this wasabi is grated, it is extracted with an ethyl acetate solvent, and this extract is dehydrated with anhydrous sodium sulfate and then concentrated with an evaporator. ⁇ -methylsulfinylalkylisothiocyanate is obtained.
  • This method is particularly suitable for the extraction of 6-methylsulfinylhexyl isothiocyanate.
  • Commercially available 6-methylsulfinylhexyl isothiocyanate may be used, and examples thereof include Wasabi sulfinyl (registered trademark) (6-MSITC (registered trademark)) manufactured by Kinshi Co., Ltd.
  • the above-mentioned extraction liquid is refine
  • drying means such as hot air drying and freeze drying may be combined.
  • ⁇ -methylsulfinylalkyl isothiocyanate specifically includes allyl isothiocyanate, secondary butyl isothiocyanate, 3-butenyl isothiocyanate, 4-pentenyl isothiocyanate, 5-hexenyl isothiocyanate, 5- Examples thereof include methylthiopentyl isothiocyanate, 6-methylthiohexyl isothiocyanate, 7-methylthioheptyl isothiocyanate, and 8-methylthiooctyl isothiocyanate.
  • components other than ⁇ -methylsulfinylalkylisothiocyanate may be further contained, and these components may be synthesized by various chemical synthesis methods in addition to extraction from a plant by the above-described method.
  • One skilled in the art can synthesize these components by methods well known in the art.
  • physiologically acceptable salt means a salt that retains a physiological effect and also retains the properties of a free base or free acid that is physiological or otherwise undesirable. To do.
  • physiologically acceptable salt include pharmaceutically acceptable salts.
  • the aging inhibitor of the present invention contains ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof as an active ingredient, and can suppress aging.
  • “suppression of aging” means, for example, suppressing physiological changes in the body accompanying aging, such as delaying aging, aging-related diseases or symptoms of the body Includes improving the condition.
  • Examples of the physiological change of the body accompanying aging include a decrease in physical ability including a decrease in athletic ability. Since the aging inhibitor of this invention delays aging, it can be said that it is an aging retarder, for example.
  • a disease or symptom associated with aging means, for example, a disease or symptom that develops with aging, and specifically emphysema (including those caused by aging); chronic obstruction Pulmonary disease (COPD); osteoporosis; Alzheimer; retinal degeneration; arteriosclerosis such as Menkeberg-type arteriosclerosis; growth disorder; early death; decreased bone density; marked atrophy of ovary, uterus and testis; Marked atrophy of the thymus; calcification of the soft tissue such as the stomach wall, skin, trachea, heart valve, arterial media; thickening of the intima of arteries; skin atrophy; loss of hair roots; loss of subcutaneous fat; Decreased ability; abnormal walking; decreased growth hormone secretory granules.
  • COPD chronic obstruction Pulmonary disease
  • osteoporosis Alzheimer
  • arteriosclerosis such as Menkeberg-type arteriosclerosis
  • growth disorder early death
  • decreased bone density marked atrophy of ovary, uterus and test
  • the aging inhibitor of the present invention can suppress, for example, activation of Calpain-1. Therefore, the antiaging agent of the present invention can improve the body condition related to a disease or a symptom caused by activation of Calpain-1, for example.
  • the disease or symptom caused by the activation of Calpain-1 may be caused by the activation of Calpain-1, for example, the disease or symptom described above.
  • the calcification inhibitor for soft tissues of the present invention contains ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof as an active ingredient, and can suppress calcification of soft tissues.
  • “calcification of soft tissue” means, for example, a phenomenon in which calcium salt is deposited or a deposited state in soft tissue.
  • soft tissue means, for example, a supporting tissue other than the skeleton in a living body, and includes, for example, connective tissue, blood vessels, striated muscle, smooth muscle, peripheral nerve tissue, and the like.
  • “calcification of soft tissue” can be called “ectopic calcification” and includes calcification in blood vessel walls, internal organs (eg, heart, lung, kidney, liver, etc.). .
  • the soft tissue calcification inhibitor of the present invention can suppress calcification of the soft tissue, for example, it improves the body condition related to a disease or symptom caused by calcification (ectopic calcification) of the soft tissue. it can.
  • the disease or symptom caused by calcification of the soft tissue may be caused by calcification of the soft tissue (ectopic calcification), for example, emphysema (including those caused by smoking), arteries Examples include sclerosis (including Menkeberg-type arteriosclerosis and arteriosclerosis associated with artificial dialysis), cirrhosis, and the like.
  • the soft tissue calcification inhibitor of the present invention like the aging inhibitor of the present invention, can suppress, for example, activation of Calpain-1, and is associated with diseases or symptoms caused by the activation of Calpain-1. You can improve your physical condition.
  • the lung tissue destruction inhibitor of the present invention contains ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof as an active ingredient, and can inhibit lung tissue destruction.
  • “suppression of lung tissue destruction” means, for example, inhibition, reduction, delay, etc. of destruction of tissues related to the lung including the alveoli.
  • the alveoli partitioning the alveoli from the alveoli Includes inhibition, reduction, delay, etc. of wall destruction.
  • Lung tissue destruction includes, for example, due to the action of Calpain-1.
  • the lung tissue destruction inhibitor of the present invention can inhibit lung tissue destruction, it can improve, for example, the state of the body related to a disease or symptom caused by lung tissue destruction.
  • the disease or symptom caused by lung tissue destruction may be that the disease or symptom is caused by lung tissue destruction, and examples thereof include emphysema (including those caused by smoking).
  • the lung tissue destruction inhibitor of the present invention like the aging inhibitor of the present invention, can suppress, for example, the activation of Calpain-1, and is associated with a disease or symptom caused by the activation of Calpain-1. The condition can be improved.
  • the aging inhibitor of the present invention can be blended in foods and drinks (functional display foods, specified health foods), pharmaceuticals, and cosmetics.
  • One embodiment of the present invention is a food / beverage product (functional indication food, specified health food), a pharmaceutical product, and a cosmetic that contain an aging inhibitor as an active ingredient and / or additive.
  • Preferred embodiments of the present invention include internal medicines (including quasi-drugs for internal use) and foods and drinks containing the aging inhibitor of the present invention.
  • sweeteners When blending the aging inhibitor of the present invention in foods and drinks, in addition to the aging inhibitor of the present invention, sweeteners, colorants, preservatives, thickeners, stabilizers, gelling agents, glues, antioxidants , Coloring agent, Bleaching agent, Antifungal agent (Antifunger), Yeast food, Gum base, Fragrance, Acidulant, Seasoning, Emulsifier, pH adjuster, Kansui, Swelling agent, Nutrition enhancer, Other food and beverage materials, etc. What is necessary is just to mix and prepare in a desired form.
  • the form of the food or drink containing the aging inhibitor of the present invention is not particularly limited.
  • supplement-type foods such as gels, granules, fine granules, capsules, tablets, powders, liquids, semi-solids; carbonated drinks, soft drinks, milk drinks, alcoholic drinks, fruit juice drinks, teas, nutrition drinks, etc.
  • Powdered beverages such as powdered juice and powdered soup; sweets such as gums, tablets, candy, cookies, gummi, rice crackers, biscuits and jellies; breads, noodles, cereals, jams, seasonings and the like.
  • These foods can be used as nutraceuticals such as dietary supplements, functional foods, foods for specified health use, foods for the sick, etc., in addition to general food and drink.
  • the antiaging agent of the present invention When blending the antiaging agent of the present invention with pharmaceuticals (including quasi-drugs), in addition to the antiaging agent of the present invention, other medicinal ingredients, pharmaceutically acceptable carriers and additions as necessary You may mix
  • pharmaceutically acceptable carriers and additives include binders, disintegrants, lubricants, wetting agents, buffers, preservatives, and fragrances.
  • the form of the pharmaceutical containing the aging inhibitor of the present invention is not particularly limited. Examples include injections, external preparations, inhalants, suppositories, films, troches, solutions, powders, tablets, granules, capsules, syrups, eye drops, eye wash, nasal drops, and the like.
  • the form suitable for oral administration namely, pharmaceutical for internal use
  • a troche a liquid, a powder, a tablet, a granule, a capsule, a syrup, etc.
  • These drugs are used as drugs for inhibiting aging.
  • a pharmaceutically or cosmetically acceptable carrier Water, oil component, etc.
  • a pharmaceutically or cosmetically acceptable carrier Water, oil component, etc.
  • the form of the cosmetic is not particularly limited as long as it can be applied to the skin.
  • liquid, emulsion, powder, solid, suspension, cream, ointment, mousse, granule, tablet, gel, jelly, paste, gel, aerosol, spray, Examples include liniments and packs. These cosmetics are used as cosmetics having an antiaging effect.
  • the aging inhibitor of the present invention can be used as an additive to foods and drinks, pharmaceuticals (including quasi drugs) or cosmetics, and foods and beverages and pharmaceuticals containing the aging inhibitor of the present invention (Pharmaceuticals) According to the cosmetics (including external products), the effects resulting from the aging inhibitor of the present invention can be obtained.
  • An aging inhibitory action can be imparted by blending the aging inhibitor of the present invention into a food, drink, pharmaceutical (including quasi-drugs) or cosmetics.
  • the anti-aging agent of the present invention can be used for imparting an anti-aging function to pharmaceuticals for internal use (including quasi-drugs for internal use) and foods and drinks.
  • the form of food and drink, pharmaceuticals (including quasi-drugs), cosmetics and the like to be blended are not limited. Including quasi-drugs) and cosmetic forms.
  • the amount of the aging inhibitor of the present invention in foods and drinks, pharmaceuticals (including quasi-drugs), and cosmetics is not particularly limited. Depending on the sex, age, food / beverage products, pharmaceuticals (including quasi-drugs) or cosmetics, the administration or intake method and frequency of these, the preference, etc., are set as appropriate. Therefore, the blending amount of the aging inhibitor of the present invention into foods and drinks, pharmaceuticals (including quasi drugs) or cosmetics is not limited. For example, the aging inhibitor of the present invention has an application amount per day for an adult.
  • the ⁇ -methylsulfinylalkylisothiocyanate may be blended so that the total amount is 0.1 to 100 mg, preferably 0.1 to 70 mg, more preferably 0.5 to 50 mg, particularly preferably 0.5 to 30 mg. Illustrated.
  • the ⁇ -methylsulfinylalkyl isothiocyanate is rapeseed such as Japanese horseradish, horseradish, cabbage, watercress, Brussels sprouts, cauliflower, radish, leach radish, rapeseed, broccoli, Takana, mustard, turnip, and Chinese cabbage.
  • these components can be obtained by extraction / purification treatment from plants, including extraction and purification treatment from family plants, and the extract itself obtained in this process may be used as the aging inhibitor of the present invention.
  • the extract is used as an applied amount per day for an adult with respect to foods and drinks, pharmaceuticals (including quasi-drugs), and cosmetics.
  • the soft tissue calcification inhibitor of the present invention can be incorporated into foods and drinks (functional display foods, specified health foods), pharmaceuticals, and cosmetics in the same manner as the aging inhibitor of the present invention.
  • One embodiment of the present invention is a food / beverage product (functional display food, specified health food), a pharmaceutical product, and a cosmetic, which contain a soft tissue calcification inhibitor as an active ingredient and / or additive.
  • Preferred embodiments of the present invention include internal medicines (including quasi-drugs for internal use) and foods and drinks containing the soft tissue calcification inhibitor of the present invention.
  • the lung tissue destruction inhibitor of the present invention can be added to foods and drinks (functional display foods, specified health foods), pharmaceuticals, and cosmetics in the same manner as the aging inhibitors of the present invention. Further, one embodiment of the present invention is a food or drink (functional indication food, specified health food), a pharmaceutical, or a cosmetic containing a lung tissue destruction inhibitor as an active ingredient and / or additive.
  • Preferred embodiments of the present invention include internal medicines (including quasi-drugs for internal use) and foods and drinks containing the tissue destruction inhibitor of the present invention.
  • the antiaging agent of the present invention can be used, for example, for prevention and / or treatment of diseases or symptoms related to aging.
  • the disease or symptom related to aging is, for example, as described above.
  • the antiaging agent of the present invention is particularly suitable for use in the prevention and / or treatment of emphysema or arteriosclerosis.
  • the aging inhibitor and the like of the present invention can suppress the activation of Calpain-1 as described above, it can be used, for example, for the prevention and / or treatment of diseases or symptoms caused by the activation of Calpain-1. it can.
  • the diseases or symptoms resulting from the activation of Calpain-1 are, for example, as described above.
  • the calcification inhibitor for soft tissues of the present invention can be used for the prevention and / or treatment of diseases or symptoms related to aging, like the aging inhibitor of the present invention.
  • the calcification inhibitor for soft tissue according to the present invention can suppress calcification (ectopic calcification) of soft tissue, for example, due to calcification (ectopic calcification) of soft tissue.
  • the disease or symptom caused by calcification of soft tissue (ectopic calcification) is, for example, as described above.
  • the lung tissue destruction inhibitor of the present invention can be used for the prevention and / or treatment of diseases or symptoms associated with aging, like the aging inhibitor of the present invention.
  • the lung tissue destruction inhibitor of the present invention can inhibit lung tissue destruction as described above, it can be used, for example, for prevention and / or treatment of diseases or symptoms caused by lung tissue destruction.
  • the disease or symptom caused by lung tissue destruction is, for example, as described above.
  • products related to the antiaging agent of the present invention can be displayed as “anti-aging”, “prevention of diseases associated with aging”, “keep youthful”, or the like, and can also be displayed in a similar manner.
  • the calcification inhibitor for soft tissue of the present invention can be used for “suppressing adverse effects due to smoking”, “suppressing adverse effects in artificial dialysis”, or “suppressing abnormal lime”.
  • ⁇ Aging inhibitory effect of wasabi-containing product 6-MSITC on aging model mice ⁇ -klotho KO mice> ⁇ -klotho KO mice born by crossing between ⁇ -klotho heterozygous mice were used.
  • ⁇ -klotho KO was determined by PCR by collecting a part of the tail from a mouse immediately before 3 weeks of age, performing genetyping. The breeding was carried out at a light / dark cycle of 8: 00-20: 00, and food and water were freely consumed. 41.7 ppm of 6-MSITC was administered as a mixed water. As a control group, water was drunk. After anesthesia of mice 4-5 weeks after 6-MSITC administration, reflux fixation was performed with 4% PFA from the heart.
  • ⁇ Effects of pulmonary emphysema and ectopic calcification of wasabi-containing 6-MSITC Wild type mouse and ⁇ -klotho KO mouse> ⁇ -klotho KO mice and Wild type mice that were born by mating between ⁇ -klotho heterozygous mice were used.
  • a genotype was determined by collecting a part of the tail from a mouse immediately before 3 weeks of age and performing genotyping.
  • ⁇ -klotho KO mice and Wild type mice were treated and paraffin sections were prepared, H & E staining, and Kossa staining were performed in the same manner as in Example 1 except that 6-MSITC administration was performed for 3 to 4 weeks.
  • paraffin sections Preparation of paraffin sections, H & E staining, and Kossa staining were performed as follows.
  • the embedded paraffin section was sliced to 4 ⁇ m with a microtome (RM2235, Leica BIOSSYSTEMS), attached to a slide glass, and incubated at 40 ° C. for 1-2 hours.
  • Paraffin sections were immersed in xylene for 3 minutes for 10 minutes, 3 times for 10 minutes in 100% ethanol, 1 for 10 minutes in 90% ethanol, 1 for 10 minutes in 80% ethanol, Deparaffinization was performed by immersing in a 70% ethanol tank once for 10 minutes.
  • FIGS. 2-1 and 2-2 The results of histological analysis of lungs and kidneys of Wild type mice and ⁇ -klotho KO mice administered with water or 6-MSITC are shown in FIGS. 2-1 and 2-2, respectively.
  • emphysema As the phenotype of ⁇ -klotho KO mice at the age of 6-7 weeks, emphysema, renal calcification, etc. are observed.
  • FIG. 2-2 From the results of Kossa staining to detect calcification (FIG. 2-2), 6-MSITC In the ⁇ -klotho KO mice to which was administered, calcification of the kidney was suppressed.
  • the brown part indicated by the arrow indicates calcification.
  • H & E staining From the results of H & E staining (FIG.
  • ⁇ Wasabi-containing material 6-MSITC suppresses Calpain-1 activation: ⁇ -klotho KO mouse> ⁇ -klotho KO mice were used as a model mouse for the pathogenesis of human aging-like symptoms. The breeding was carried out at a light / dark cycle of 8: 00-20: 00, and food and water were freely consumed. ⁇ -klotho hetero-deficient mice self-bred, tails were collected just before 2 weeks of age, and ⁇ -klotho KO mice were selected by Genotyping using the PCR method. Two-week-old male ⁇ -klotho KO mice were administered intraperitoneally with 6-MSITC 5 mg / kg for 6 consecutive days.
  • physiological saline (Otsuka Pharmaceutical Co., Ltd., Otsuka raw food injection) was administered.
  • 50 times the lung weight of the protease inhibitor cocktail containing RIPA buffer (Nacalai Tesque, model number: 08714-04) was added and homogenized with a biomasher SP (NIPPI, model number: 893163).
  • biomasher SP NIPPI, model number: 893163
  • the mixture was incubated on ice for 30 minutes, centrifuged at 16,000 rpm, 4 ° C., 10 minutes, and the supernatant was collected. Protein quantification was performed with Pierce BCA protein assay kit (THERMO FISHER SCIENTIFIC, model number: 23227).
  • a lung tissue extract obtained by adding a sample buffer and incubating at 95 ° C. for 5 minutes was used as a western blot sample. Protein separation was performed by SDS-PAGE on 7.5% or 4-20% gradient gels. After transferring the membrane, blocking (washing for 5 minutes ⁇ 3), 5% skim milk for 1 hour, washing (10 minutes ⁇ 2, 5 minutes ⁇ 1), Anti-Calpain-1 large subantibody as the primary antibody (dilution ratio 1: 1000, CELL SIGNALING TECHNOLOGY, model number: # 2556), Anti-Calpastatin antibody (dilution ratio 1: 1000, CELL SIGNALING TECHNOLOGY, model number: # 4146), Anti-Alpha fodrin antibody A1: B1000, B1: C1 Anti-b-actin antibody (dilution ratio 1: 1000, CELL SIGNALING TECHNOLOGY, model number: # 49 Using 7), 4 °C, it reacted at overnight.
  • ECL-anti-rabbit IgG HRP antibody (dilution ratio 1: 10000, GE HEALTHCARE, model number: LNA934V / AG) is used as the secondary antibody at room temperature. Reacted for hours. After washing (10 minutes ⁇ 3) and reaction with ECL Prime Detection Reagent (GE HEALTHCARE, model number: RPN2236), a band was detected with Fuji Medical Film Processor FPM100.
  • Calpain-1 activation in lung tissue was evaluated by analyzing the band pattern in Western blot for the lung tissue extract. As shown in FIG. 3, when comparing the ⁇ -klotho KO / Control group and the ⁇ -klotho KO / 6-MISITC group, the ⁇ -klotho KO / 6-MSITC group is compared to the ⁇ -klotho KO / Control group.
  • the cytoskeletal protein ⁇ -II spectrin also known as ⁇ -fodrin
  • ⁇ Wasabi-containing material 6-MSITC suppresses Calpain-1 activation: Wild type mouse and ⁇ -klotho KO mouse> ⁇ -klotho KO mice and Wild type mice that were born by mating between ⁇ -klotho heterozygous mice were used. Tail was collected just before 2 weeks of age, and ⁇ -klotho KO mice and Wild type mice were selected by Genotyping using PCR method. In the same manner as in Example 3, ⁇ -klotho KO mice and Wild type mice were treated, and each lung tissue extract was analyzed by Western blot.
  • FIG. 4 shows the result of Western blot.
  • FIG. 4A is an image showing a band pattern of a western blot.
  • FIGS. 4B and 4C are graphs showing the ratio of the cleavage band intensity to the intact band intensity for Calpain-1 and ⁇ -II spectrin.
  • the band intensity was quantified by ImageJ, which is image analysis software. As shown in FIGS.
  • Beam balance test Beam balance test
  • a bar 24 cm in diameter and 95 cm in length was installed parallel to the ground at a position 50 cm above the ground, a retreat place was set up as a goal point on one end of the bar, and a start point was set on the opposite side .
  • the exercise ability was evaluated by measuring the time required for the mouse to move 80 cm from the start point. Each mouse was tested three times, and the average value was taken as the running time value of the mouse.
  • the mouse used the mouse trained for several days so that it may move in advance from a starting point to an evacuation place.
  • Table 1 and FIG. 5 show the results of a beam balance test for evaluating cooperative movement, muscle strength, and balance.
  • Table 1 and FIG. 5 show the results of a beam balance test for evaluating cooperative movement, muscle strength, and balance.
  • Table 1 and FIG. 5 show the results of a beam balance test for evaluating cooperative movement, muscle strength, and balance.
  • 6-MSITC suppresses the progress of aging because the decrease in athletic ability is a phenomenon that occurs remarkably with aging.
  • SAMP1 mouse> SAM (Sensence-Accelerated Mouse) P1 was used as an aging mouse model. The breeding was carried out at a light / dark cycle of 8: 00-20: 00, and food and water were freely consumed. 17-week-old SAMP1 mice were purchased from Japan SLC, and 20-week-old SAMP1 mice were mixed with 41.7 PPM of 6-MSITC (6-MSITC group). As the Control group, water was administered. After appearance evaluation of mice at 77 weeks of age, reflux fixation was performed with 4% PFA from the heart.
  • the collected organs were immersed and fixed with 4% PFA (4 ° C., overnight), washed with PBS, paraffin sections were prepared, and H & E staining was performed. Preparation of paraffin sections and H & E staining were performed as follows. First, the embedded paraffin section was sliced to 4 ⁇ m with a microtome (RM2235, Leica BIOSSYSTEMS), attached to a slide glass, and incubated at 40 ° C. for 1-2 hours.
  • a microtome R2235, Leica BIOSSYSTEMS
  • Paraffin sections were immersed in xylene for 3 minutes for 10 minutes, 3 times for 10 minutes in 100% ethanol, 1 for 10 minutes in 90% ethanol, 1 for 10 minutes in 80% ethanol, Deparaffinization was performed by immersing in a 70% ethanol tank once for 10 minutes. The section after deparaffinization was washed with water, immersed in Mayer's hematoxylin solution for 10 minutes, lightly washed with water at 37 ° C, and immersed in water at 37 ° C for 5 minutes. It was soaked in 70% ethanol and soaked in 1% eosin lees for 1 minute 30 seconds. After performing 10 intrusions with 100% ethanol slag 4 times, 10 infestations with xylene slags were performed 4 times. Sections that had been penetrated with xylene were sealed with marinol (Mudo Chemical, product number 20009) and observed with a microscope.
  • marinol Modo Chemical, product number 20009
  • FIG. 6 and FIG. 7 show the external appearance of the skin of the SAMP1 mouse administered with 6-MSITC and the results of histological analysis.
  • SAMP1 mice have wrinkles around the eyes as the age progresses.
  • 3 out of 3 mice in the SAMP1 / Control group showed wrinkles, but 1 out of 2 mice in the SAMP1 / 6-MSITC group showed wrinkles and no wrinkles.
  • the appearance of the mice was normal.
  • the histological findings of the skin maintained the thickness and structure of the adipose tissue in the SAMP1 / 6-MSITC group mice as compared with the SAMP1 / Control (Vehicle) group mice. These results indicate that 6-MSITC suppresses the progress of aging.
  • C57 / BL10-mdx mice were used as disease model mice for Duchenne muscular dystrophy.
  • C57BL / 10 mice which are genetic backgrounds, were used as Wildtype mice. The breeding was carried out at a light / dark cycle of 8: 00-20: 00, and food and water were freely consumed.
  • mdx mice self-bred, and C57BL / 10 mice were purchased from Japan SLC.
  • Four-week-old male mdx mice and male C57BL / 10 mice were administered 125.1 ppm 6-MSITC in mixed water for 4 weeks. As a control group, water was drunk.
  • a RIPA buffer Nacalai Tesque, model number: 08714-04
  • a biomasher SP Nappi, model number: 893163
  • the mixture was incubated on ice for 30 minutes, centrifuged at 16,000 rpm, 4 ° C., 10 minutes, and the supernatant was collected.
  • Protein quantification was performed with Pierce BCA Protein Assay Kit (THERMO FISHER SCIENTIFIC, model number: 23227). After protein quantification, a muscle extract obtained by adding a sample buffer and incubating at 95 ° C. for 5 minutes was used as a western blot sample.
  • Protein separation was performed by SDS-PAGE on 7.5% or 4-20% gradient gels. After transferring the membrane, blocking (washing for 5 minutes ⁇ 3), 5% skim milk for 1 hour, washing (10 minutes ⁇ 2, 5 minutes ⁇ 1), Calpain-1 Large subunit antibody as the primary antibody (dilution ratio 1: 1000, CELL SIGNALING) TECHNOLOGY, model number: # 2556), calpastatin antibody (dilution ratio 1: 1000, CELL SIGNALING TECHNOLOGY, model number: # 4146), b-actin antibody (dilution ratio 1: 1000, CELL SIGNALING TECHNOLOG67, model number: The reaction was performed at 4 ° C. and overnight.
  • ECL-anti-rabbit IgG HRP antibody (dilution ratio 1: 10000, GE HEALTHCARE, model number: LNA934V / AG) is used as the secondary antibody at room temperature. Reacted for hours. After washing (10 minutes ⁇ 2, 5 minutes ⁇ 1) and reaction with ECL Prime Detection Reagent (GE HEALTHCARE, model number: RPN2236), the band was detected with Fuji Medical Film Processor FPM100.
  • An aging inhibitor, a soft tissue calcification inhibitor, and a lung tissue destruction inhibitor containing ⁇ -methylsulfinylalkylisothiocyanate or a physiologically acceptable salt thereof according to the present invention are used in foods, beverages, pharmaceuticals or cosmetics. Can be used.

Abstract

La présente invention concerne : un nouvel inhibiteur de vieillissement, un inhibiteur de calcification de tissu mou et un inhibiteur de destruction de tissu pulmonaire ; des produits alimentaires et de boisson, des produits pharmaceutiques et des produits cosmétiques qui comprennent lesdits inhibiteurs ; et un procédé d'utilisation desdits produits. L'inhibiteur de vieillissement, l'inhibiteur de calcification de tissu mou et l'inhibiteur de destruction de tissu pulmonaire contiennent de l'isothiocyanate de ω-méthylsulfinylalkyle ou un sel physiologiquement acceptable de ce dernier.
PCT/JP2017/036547 2016-10-07 2017-10-06 Inhibiteur de vieillissement, inhibiteur de calcification de tissu mou et inhibiteur de destruction de tissu pulmonaire WO2018066707A1 (fr)

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