JP2016169168A - Heat shock protein expression inducer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel natural product-derived component which can induce the expression of heat shock protein.SOLUTION: A heat shock protein expression inducer comprises a Monascus dye as an active ingredient.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a heat shock protein expression inducer.

  As one of the defense systems of living bodies, for example, when it is subjected to various stresses such as high temperature exceeding the general physiological temperature of 3 ° C or more, various chemicals, heavy metals, radiation, ultraviolet rays, starvation, oxygen deficiency, etc. It is known that the expression (production) of a specific protein called heat shock protein (hereinafter also referred to as “HSP”) is induced in order to protect cells from heat. There are several types of HSP, depending on the molecular weight, for example, HSP90 family (molecular weight 90 kDa or more and 110 kDa or less), HSP70 family (molecular weight 70 kDa or more and less than 80 kDa), HSP60 family (molecular weight 60 kDa or more and less than 70 kDa) and low molecular weight HSP. It is classified as a family (molecular weight less than 60 kDa).

  HSP plays a role in helping to restore a physiological function when a protein in a cell is denatured by various stresses as described above, whereby the protein again forms a normal three-dimensional structure (folding). Moreover, even under non-stress, it exists in a certain amount in the living body, and contributes to the defense of the living body and the maintenance of the homeostasis, such as assisting a protein having a problem with folding. Therefore, if the expression of HSP having such a function is intentionally induced, it results from protection of cells from various stresses, acceleration of recovery of physiological functions of cells that have deteriorated or malfunctioned, and abnormal protein folding. The effect of prevention or treatment of various diseases and symptoms is expected. Specifically, by inducing the expression of HSP, for example, prevention or treatment of cancer, gastric ulcer, brain diseases such as cerebral infarction and Alzheimer's disease, ulcerative colitis, pulmonary fibrosis, wound healing promotion, anti-inflammatory, It is known to be effective in antioxidant, melanin production inhibition, anti-wrinkle and the like.

  As a method for inducing the expression of HSP, for example, stress is intentionally applied to a living body such as thermotherapy. However, in this method, there is a risk that an excessive burden is applied to the living body if the stress applied is too strong. Therefore, a method for inducing the expression of HSP in a living body without depending on such external stress has been proposed.

  Particularly in recent years, for example, among the four characteristics of herbal medicine, it contains heat shock protein inducer (Patent Document 1), Arnica, which contains a plant extract that is the origin of herbal medicine having properties corresponding to warm or flat. At least one selected from the group consisting of heat shock protein expression inducer (Patent Document 2), zerumbone and its analogs, carotene, lycopene, zeaxanthin, cryptoxanthin, lutein, phenylethylisothiocyanate, and ursolic acid Heat shock protein 70 production comprising, as an active ingredient, a heat shock protein expression inducer (patent document 3) characterized by containing as an active ingredient, a yeast cell extract cultured in a medium containing 10 mM or more of a zinc compound Thermal sheath containing an inducer (Patent Document 4), kale or processed product thereof as an active ingredient Kkutanpaku protein expression promoting agent (Patent Document 5) or the like, a method of utilizing the components derived from highly safe natural products has attracted attention. In such a trend, there has been a demand for a component derived from a novel natural product that can induce the expression of HSP.

JP 2008-127296 A JP 2011-190200 A WO2012 / 043808 JP 2014-108945 A JP 2014-088378 A

  An object of the present invention is to provide a novel natural product-derived component that can induce the expression of a heat shock protein.

  As a result of intensive studies to solve the above-mentioned problems, the present inventor has found that a beech mushroom pigment, which is used as a natural colorant for foods and has abundant edible experience, induces the expression of HSP. Invented the invention.

  That is, this invention consists of the heat shock protein expression inducer which uses Benikouji pigment | dye as an active ingredient.

The heat shock protein expression inducer of the present invention induces the expression of HSP, particularly HSP70, in a living body.
The heat shock protein expression inducer of the present invention can be used as a preventive or therapeutic agent for various diseases related to the action of HSP.
Since the heat shock protein expression inducer of the present invention comprises a beech mushroom dye approved as a food additive as an active ingredient, it can be safely administered to a living body both orally and parenterally.

The graph which shows the measurement result of HSP70 content in each test section.

  Benikouji pigment, which is an active ingredient of the heat shock protein expression inducer of the present invention (hereinafter also referred to as “HSP expression inducer”), is obtained by extraction from a culture product of Beniculomycetes (ie, filamentous fungi belonging to the genus Monascus). The main components are monascorbulins, ancaflavins, and rubropanktatins.

  There is no restriction | limiting in particular in the form of a beige mushroom pigment | dye, Any may be a bemoji pigment liquid formulation, a bemoji pigment powder formulation, etc. A liquid preparation of Benikouji pigment can be prepared by extracting Benikouji pigment from a culture product of Beniculobacterium using water or a mixed solution of water and alcohol as an extraction solvent. In addition, the bemoji dye powder preparation is prepared by mixing the bemoji dye liquid preparation with a powdered base material such as dextrin and spray-drying the mixture liquid by a conventional method, or concentrating the bemoji dye liquid preparation and vacuum freeze-drying. It can prepare by the method to do.

  Although there is no restriction | limiting in particular in the color value of a beige mushroom pigment | dye, For example, the color value of 10-200 will be used normally for a bemoji dye liquid formulation, and the color value of 100-3000 will be used for a bemoji pigment powder formulation. In addition, the color value as referred to in the present invention is measured according to “17.

  Examples of Benikouji pigment include Riquecolor R-30 (trade name; Benikouji pigment liquid formulation; color value 60; manufactured by Riken Vitamin Co., Ltd.), Riquecolor R-1000 (P) (trade name; Benikouji pigment powder formulation; color value 200; Riken Vitamin Co., Ltd.), Riquecolor R-2000 (P) (trade name; Benikouji pigment powder formulation; color value 2000; manufactured by Riken Vitamin Co., Ltd.), Riquecolor R-4200 (SD) (trade name; Benikouji pigment powder formulation; color value 2000) ; Manufactured by Riken Vitamin Co., Ltd.) and the like are commercially produced and sold, and these can be used as the HSP expression inducer of the present invention.

  The HSP expression inducer of the present invention may be used as it is, or other optional components (pharmaceutical additives, food additives, food materials, etc.) as long as they do not inhibit the effects of the present invention. ) And may be prepared as a composition containing at least a beige moji pigment.

  Examples of optional components other than the beech powder used in the preparation of the composition include excipients (lactose, dextrin, corn starch, crystalline cellulose, etc.), lubricants (magnesium stearate, sucrose fatty acid ester, glycerin fatty acid ester). Etc.), disintegrating agents (carboxymethylcellulose calcium, anhydrous calcium hydrogen phosphate, calcium carbonate, etc.), binders (starch glue solution, hydroxypropylcellulose solution, gum arabic solution, etc.), solubilizers (gum arabic, polysorbate 80, etc.) , Sweeteners (sugar, fructose, glucose liquid sugar, honey, aspartame, etc.), coloring agents (β-carotene, food tar pigment, riboflavin, etc.), preservatives (sorbic acid, methyl parahydroxybenzoate, sodium sulfite, etc.), thickening Agent (sodium alginate, carboxy Chill cellulose sodium, sodium polyacrylate, etc.), antioxidants (BHT, BHA, ascorbic acid, tocopherol, etc.), flavors (mint, strawberry flavor, etc.), acidulants (citric acid, lactic acid, DL-malic acid, etc.), Seasoning (DL-alanine, sodium 5'-inosinate, sodium L-glutamate, etc.), pH adjuster (citric acid, trisodium citrate, etc.), emulsifier or surfactant (glycerin fatty acid ester, sucrose fatty acid ester, Alkyl sulfate salts, alkyldimethylaminoacetic acid betaine, dialkyldimethylammonium chloride, etc.), oils (higher alcohols, ester oils, liquid paraffin, petrolatum, squalane, waxes, natural fats and oils, silicone oils, etc.), lower alcohols (ethanol, isopropyl) Alcohol), Monohydric alcohols (propylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, maltitol, etc.), ultraviolet absorbers (ethyl paraaminobenzoate, octyl salicylate, 2-ethylhexyl paramethoxycinnamate, 2-hydroxy-4 -Methoxybenzophenone, etc.), inorganic powder (talc, kaolin, boron nitride, etc.), organic powder (cellulose powder, cross-linked silicone powder, polytetrafluoroethylene powder, etc.), vitamins, minerals, amino acids, etc. .

  When the HSP expression inducer of the present invention is prepared as a pharmaceutical product, the dosage form is not particularly limited, and can be prepared into an arbitrary dosage form according to the administration route. Examples of dosage forms suitable for oral administration include tablets, capsules, powders, fine granules, granules, solutions, syrups, lozenges, etc. Examples of dosage forms suitable for parenteral administration include injection. Agents, drops, suppositories, inhalants, transdermal absorbents, transmucosal absorbents, patches, emulsions, creams, ointments, lotions, poultices and the like. The injection may be used for any of intravenous injection, intramuscular injection, subcutaneous injection, infusion and the like.

  The content of the beige mushroom pigment in 100% by mass of the pharmaceutical varies depending on its color value, pharmaceutical dosage form, administration route, etc., but is usually 0.0001 to 50% by mass, preferably 0.001 to 20% by mass, more preferably Can be set within a range of 0.01 to 10% by mass.

  Moreover, the HSP expression inducer of the present invention can be used by adding to foods and drinks, cosmetics and the like.

  When the HSP expression inducer of the present invention is added to a food or drink, the form of the food or drink to be added is not particularly limited. For example, soft drink, drop, candy, chewing gum, chocolate, gummi, yogurt, ice Examples include cream, pudding, jelly confectionery, and cookies.

  Although the amount of Benikouji pigment added to the food and drink varies depending on the color value and the form of the food and drink, etc., the range is usually 0.001 to 5% by weight, preferably 0.01 to 3% by weight in 100% by weight of the food and drink. Can be set within.

  When the HSP expression inducer of the present invention is used by being added to a cosmetic, there is no particular limitation on the form of the cosmetic to be added, but skin cosmetics such as lotions, emulsions, creams, cosmetics, packs, etc. Makeup base lotion, foundation cosmetics such as makeup base cream, foundations of various dosage forms such as emulsion, oily, solid, makeup cosmetics such as eye color and teak color, hand cream, leg cream, neck Examples include body cosmetics such as creams and body lotions.

  Although the amount of Benikouji pigment added to the cosmetic varies depending on the color value, the form of the cosmetic, and the like, it is usually in the range of 0.001 to 10% by mass, preferably 0.01 to 5% by mass in 100% by mass of the cosmetic. Can be set within.

  The HSP expression inducer of the present invention can induce the expression of HSP in vivo by being orally or parenterally administered to a living body as a pharmaceutical product or added to food or drink, cosmetics, etc. as described above. it can. In addition, it is preferable to administer orally since the beige moji pigment | dye which is an active ingredient of the HSP expression inducer of this invention is a highly safe component approved as a food additive.

  The HSP that induces expression of the HSP expression inducer of the present invention is not particularly limited. For example, those belonging to the HSP90 family (molecular weight 90 kDa to 110 kDa; HSP90α, HSP90β, HSP104, HSP110 etc.), those belonging to the HSP70 family (molecular weight) 70 kDa or more and less than 80 kDa; HSP70, HSP72, HSP73 etc., those belonging to HSP60 family (molecular weight 60 kDa or more and less than 70 kDa; HSP60, HSP65 etc.) and those belonging to low molecular weight HSP family (molecular weight less than 60 kDa; HSP47, HSP40, HSP27, HSP20 Etc.). Among these, the HSP expression inducer of the present invention exhibits an excellent expression inducing effect against those belonging to the HSP70 family, particularly HSP70 having a molecular weight of 70 kDa.

  The HSP expression inducer of the present invention induces the expression of the above HSP, thereby protecting cells from various stresses such as high temperature, various chemicals, heavy metals, radiation, ultraviolet rays, starvation, oxygen deficiency, It contributes to the promotion of the recovery of physiological functions of cells that have malfunctioned, the prevention or treatment of diseases and symptoms caused by abnormal protein folding, and the like. Specifically, for example, cancer, gastric ulcer, cerebral disease such as cerebral infarction and Alzheimer's disease, ulcerative colitis, pulmonary fibrosis and the like preventive or therapeutic agent, wound healing promoter, anti-inflammatory agent, antioxidant, melanin production It can be used as an inhibitor, an anti-wrinkle agent and the like.

  The dose of the HSP expression inducer of the present invention to a living body varies depending on the composition of the HSP expression inducer, the purpose of administration, the administration route, and the like. For example, in the case of oral administration, In the range of 0.01 to 600 mg per day for adults.

  Hereinafter, the present invention will be described more specifically based on examples, but the present invention is not limited thereto.

[HSP70 expression induction test]
(1) Cell culture Human cervical cancer cells (HeLa cells; RCB0007; RIKEN BioResource Center) were seeded in a 24-well plate at a cell concentration of 5 × 10 ⁵ cells / 0.5 mL, and 10 vol% FBS and The cells were cultured overnight in a DMEM medium containing 1% by volume penicillin-streptomycin (Nacalai Tesque) at 37 ° C. and 5% CO ₂ . Thereafter, the fresh medium containing Benikouji pigment (trade name: Riquecolor R-4200; Benikouji pigment powder preparation; color value 2000; manufactured by Riken Vitamin Co., Ltd.) was replaced with a prescribed concentration, and cultured under the same conditions for another 24 hours ( Test sections 1-3). In addition, as a control, a test group (test group 4; negative control) that was treated in the same manner after changing to a medium containing no beech powder, and a medium that did not contain bemoji dye were exchanged, and 17 hours after the medium was changed. A test group (test group 5; positive control) subjected to heat stress at 42 ° C. for 1 hour was provided. Table 1 shows the concentration of the beetle pigment and the presence or absence of heat stress in each test section.

(2) Evaluation of HSP70 expression induction After the cell culture in the above test groups 1 to 5, the supernatant medium was removed, the cells were peeled from the wells with trypsin, and ice-cold PBS (Nissui Pharmaceutical Co., Ltd.) was added and washed. After that, the cells were collected. Extraction buffer containing protease inhibitor attached to HSP70 ELISA kit (manufactured by Stress Xpress) was added to the collected cells, the cells were disrupted using vortex, and centrifuged at 12000 × g for 10 minutes to obtain the supernatant as a protein. It recovered as an extract.
About each obtained protein extract, protein concentration was measured according to the kit predetermined | prescribed technique using Micro BCA Protein Assay kit (made by PIERCE Biotechnology), and the total protein amount in this protein extract was calculated | required. On the other hand, using the HSP70 ELISA kit, the HSP70 content in each protein extract was measured according to the kit predetermined method, and the HSP70 content per 1 μg of protein in the cultured cells of each test section was calculated. The results are graphed and shown in FIG.

  As is clear from the results in FIG. 1, the HSP content in the cultured cells in the test group 5 as a positive control was higher than that in the test group 4 as a negative control. In the cultured cells of the test groups 1 to 3, an increase in the HSP70 content was observed as compared to the test group 4. From this, it was found that the beech mushroom dye induces intracellular HSP expression.

Claims (1)

  A heat shock protein expression inducer comprising Benikouji pigment as an active ingredient.

Images