JP2015091780A - Hyaluronidase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a formulation which inhibits hyaluronidase, and is useful for the treatment of skin wrinkles and sagging as well as contact dermatitis, psoriasis, pemphigus vulgaris and the like.SOLUTION: The inventors have found out that extracts of Atractylodis rhizoma, Ishige, a mulberry looper, and Manchurian wild rice serve the purpose of this invention.

Description

  The present invention relates to a preparation that inhibits hyaluronidase and is effective against wrinkles and tarmi, and further against contact dermatitis, psoriasis, pemphigus vulgaris, and the like.

Typical changes in the aging of human skin are wrinkles and tarmi, which is one of the major skin problems for older people and women.
The causes include the effects of drying, oxidation, sunlight (ultraviolet rays), etc. in addition to aging.
Specific histological phenomena include changes in extracellular matrix components such as collagen, elastin, and glycosaminoglycans in the dermis. Among these glycosaminoglycans, hyaluronic acid is widely distributed in connective tissues. Known as a high molecular weight polysaccharide, it retains a large amount of moisture in the skin and has a gel-like form. Therefore, qualitative and quantitative reduction of hyaluronic acid is considered to be important in skin aging.
In addition, there are various causes and origins of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other types of skin diseases, including hyaluronidase. Is known.
The hyaluronidase is an enzyme that hydrolyzes hyaluronic acid, but hyaluronic acid is a mucopolysaccharide found in the mesenchymal tissue and serves to prevent invasion and transmission of microorganisms and toxicants. When activated, the damage caused by the decrease in the protective effect of hyaluronic acid is likely to occur. Hyaluronidase is also related to type I allergic reaction and is considered to be an enzyme that controls degranulation from mast cells. Therefore, the enhancement of moisture retention or anti-inflammatory action can be expected by stabilizing hyaluronic acid and preventing the release of various chemical mediators from mast cells.
Therefore, attempts have been made to extract such substances having hyaluronidase inhibitory activity from natural products. For example, extracts of the genus Osbecchia, Fuji tea extract, rosemary, thyme and Melissa extract have been reported. Has been. (See Patent Documents 1 to 3)

The white rhododendron is one of the herbal medicines, and the rhizome part excluding the peripheries of the genus Ochera (Atractylodes japonica), Atractylodes ovata, etc. This refers to the rhizome part of Atractylodes ovata. It is used as an external preparation for the purpose of whitening and wrinkle improvement. (Patent Documents 4 to 5).
Ishige is a seaweed of the brown algal plant gate, brown alga net, Nagamatsumo, Ishigeaceae, Ishige genus and is called the Ishige okamurai Yendo, and is a brown alga widely distributed along the coast south of Ibaraki. Not suitable for edible use. Already used for skin external preparations and foods for whitening, hair growth, slimming, antibacterial and antiseptic purposes. (Patent Documents 6 to 8)
Mulberry is a dry leaf of a mulberry tree whose scientific name is Morus Bombycis, Morus alba, etc. Morse bombibisis is distributed throughout Japan, from the Minami Kuril, Korea, China, Vietnam to India, and from Kashmir to the temperate zone to the subtropics. A deciduous tree that grows in the mountains and is planted in a field, called mulberry or yamagua. Mors Alba is native to China and the Korean peninsula, and has come to Japan since ancient times. It is a deciduous tree that is planted in various places for garden trees, street trees, and sericulture, and is called Karagua and Touguwa. Conventionally, it has been used as a medicine for antipyretic, antitussive, cool blood, clear eye medicine, cold, cough, headache, eye disease, beriberi, edema, abdominal pain, diarrhea, eradication blood, etc. It is used as a treatment for acne. (Patent Document 9)
Makomo is a perennial plant belonging to the genus Macomo, which is distributed in East Asia and Southeast Asia including Japan, and grows on watersides of rivers, river banks, lakes, and so on, and is called Zizania latifolia. Macomo's flower buds are infested with Aspergillus oryzae and the roots are enlarged in the shape of a cocoon. This is called “Makomodake”, and some green leaf sheaths are peeled off and used as food. In addition, the buds made with kurobushi are used for paints and eyebrows. In addition to food, this site is used as an external preparation for skin having anti-inflammatory action, an agent for maintaining undifferentiation of stem cells, and a proliferation promoter. (Patent Documents 10 to 11) Other parts are only partially used for fertilizers and mushiro (leaves), and an effective and advanced utilization method has been demanded.

Japanese Patent Laid-Open No. 2003-55242 JP 2003-12532 A JP-A-8-333267 JP 09-077635 A JP-A-10-194946 Japanese Patent Laid-Open No. 04-95012 Japanese Patent Laid-Open No. 11-180813 JP 2000-236846 A JP-A-10-007544 JP 2011-93880 A JP 2011-211957 A

  The object of the present invention is to inhibit hyaluronidase and to obtain a preparation effective for wrinkles and tarmi, as well as for contact dermatitis, psoriasis, pemphigus vulgaris and the like.

As a result of intensive studies by the present inventors, it has been found that extracts of white birch, ishige, mulberry branch and macomo achieve the above-mentioned purpose.
It is preferable to extract white birch, ishige, mulberry branch, and macomo after processing such as shredding, drying, and pulverization in consideration of extraction efficiency.
Drying may be performed in the sun or using a commonly used dryer.
As the solvent used for the extraction, water, a hydrophilic organic solvent, or a mixture thereof is used.
Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, as well as water that has been subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.
Examples of the hydrophilic organic solvent include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene glycol, propylene glycol, Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and a mixed solvent of these hydrophilic organic solvents and water can be used.
In addition, when using the mixed solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, it is 1-20 mass parts with respect to 10 mass parts of water, and in the case of a lower aliphatic ketone, it is 10 mass parts of water. On the other hand, it is preferable to add 1 to 15 parts by mass. In the case of a polyhydric alcohol, it is preferable to add 1 to 20 parts by mass with respect to 10 parts by mass of water.
The amount of the organic solvent used for the extraction is preferably about 5 to 100 times, more preferably about 10 to 50 times the amount of the plant (seaweed) as the raw material. Furthermore, in order to raise extraction efficiency, you may stir or homogenize in an extraction solvent. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it to about 1 hour to 14 days.
The extraction operation is not limited to a single operation. A fresh solvent can be added again to the residue after extraction to perform an extraction operation, or the extraction solvent can be brought into contact with the extraction raw material a plurality of times.
If necessary, purification treatment such as deodorization and decolorization may be further added within a range that does not affect the effect, and concentration can be performed by a vacuum concentrator such as an evaporator or solvent removal by heating.
Further, this extract can be further purified using a synthetic adsorbent (Diaion HP20, Sephabies SP825, Amberlite XAD4, MCIgelCHP20P, etc.), dextran resin (Sephadex LH-20, etc.), ultrafiltration and the like. is there.

  The preparation of the present invention exhibits drug efficacy for oral, injection, and external use, but is preferably used as a skin external preparation. Skin external preparations include skin cosmetics, external quasi-drugs, and medical skin external preparations.

  In addition to the above ingredients, the preparations of the present invention include surfactants, oily ingredients, moisturizers, polymer compounds, ultraviolet absorbers, anti-inflammatory agents, bactericides, and other agents used in various pharmaceutical and cosmetic preparations. Antioxidants, sequestering agents, preservatives, vitamins, pigments, fragrances, water and the like can be blended.

  As the surfactant, any of anionic, cationic, nonionic, natural, and synthetic surfactants can be used, but it is preferable to use a nonionic surfactant in consideration of irritation to the skin. Examples of the nonionic surfactant include glycerin fatty acid ester, propylene glycol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene sorbite fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene glycol. , Polyoxyethylene polyoxypropylene alkyl ether, polyethylene glycol fatty acid ester, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, alkylglycoside and the like.

  Examples of the oil component include fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, essential oils, silicone oils, and the like. Examples of the fats and oils include soybean oil, nutka oil, jojoba oil, avocado oil, almond oil, olive oil, cacao oil, sesame oil, persic oil, castor oil, coconut oil, mink oil, beef fat, pork fat and the like, and these Hardened oil obtained by hydrogenation of natural fats and oils and synthetic triglycerides such as myristic acid glyceride and 2-ethylhexanoic acid triglyceride; waxes include, for example, carnauba wax, whale wax, beeswax, lanolin and the like; hydrocarbons For example, liquid paraffin, petrolatum, paraffin microcrystalline wax, ceresin, squalane, bristan etc .; higher fatty acids include, for example, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, Linolenic acid, lanolinic acid, isostearic acid, etc .; Examples of the class alcohols include lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, 2-hexyldecanol, etc .; examples of the esters include cetyl octanoate, triglyceride octanoate, myristyl lactate, cetyl lactate, Examples of essential oils include mint oil, jasmine, isopropyl myristate, myristyl myristate, octyldodecyl myristate, isopropyl palmitate, isopropyl adipate, butyl stearate, decyl oleate, cholesterol isostearate, POE sorbite fatty acid ester, etc. Oil, pepper brain oil, cypress oil, spruce oil, ryu oil, turpentine oil, cinnamon oil, bergamot oil, mandarin oil, ginger oil, pine , Lavender oil, bay oil, clove oil, hiba oil, rose oil, eucalyptus oil, lemon oil, thyme oil, peppermint oil, rose oil, sage oil, menthol, cineole, eugenol, citral, citronellal, borneol, linalool, geraniol, Camphor, thymol, spirantol, pinene, limonene, terpene compounds, etc .; examples of silicone oils include dimethylpolysiloxane. These oily components described above can be used alone or in combination of two or more. In the present invention, among these, myristic acid glyceride, 2-ethylhexanoic acid triglyceride, lanolin, liquid paraffin, petrolatum, paraffin microcrystalline wax, squalane, lauric acid, myristic acid, palmitic acid, linoleic acid, linolenic acid, isostearic acid , Cetyl alcohol, stearyl alcohol, oleyl alcohol, cholesterol, cetyl octanoate, triglyceride octanoate, isopropyl myristate, octyldodecyl myristate, cholesterol isostearate, POE sorbite fatty acid ester, peppermint oil, spruce oil, cinnamon oil, rose Oil, menthol, cineol, eugenol, citral, citronellal, geraniol, pinene, limonene, dimethylpolysiloxane It is preferable to use.

The following components can be further added to the preparation of the present invention, but the components are not limited thereto.
Colors: Yellow color No. 4, Blue No. 1 and Yellow No. 202 are recognized as food additives such as tar color dyes I and II, chlorophyll, riboflavin, crocin, safflower, anthraquinone, etc. Natural pigments.
Vitamins; vitamin A, vitamin C, vitamin D, vitamin E, etc.
Others: bactericides, preservatives, other ingredients necessary for formulation.

The preparation of the present invention can be produced according to a conventional method by adding the optional components to the essential components as necessary, and can be in the form of cream, emulsion, lotion or the like.
The compounding quantity to a formulation is 0.00001 to 10 weight% as solid content, Preferably it is 0.0001 to 1.0 weight%.

Next, the present invention will be described in detail with reference to examples (production examples).

Production Example 1
Add 300 ml of purified water to 10 g of Chinese white birch (dried product) and heat for 3 hours. This was allowed to cool, then filtered and lyophilized.

Production Example 2
500 g of purified water was added to 10 g of ishige (dried product) and sufficiently homogenized. After centrifuging this, adding 4 times the amount of ethanol to the supernatant and stirring. This was centrifuged to obtain a supernatant. This was evaporated and then lyophilized.

Production Example 3
To 10 g of mulberry branches (dried product), 300 ml of 50% ethanol was added and allowed to stand for 5 days with occasional stirring. This was filtered, evaporated, and lyophilized.

Production Examples 4-6
After washing the makomo with water, it was cut into chopped mushrooms (edible part), leaves, roots, and stems, and then dried at 50 ° C. for 24 hours.
50 g of makomodake (edible part), leaves, and stems were separately taken, 800 ml of 50% ethanol aqueous solution was added, and the mixture was allowed to stand for 3 days with occasional stirring. After evaporation, it was freeze-dried.
Macadamella (edible part) 50% ethanol extract was designated as Production Example 4, leaf 50% ethanol extract as Production Example 5, and stem 50% ethanol extract as Production Example 6.

Confirmation test (hyaluronidase inhibition test)
The following method was used.
0.2 ml of 0.1 M acetate buffer (pH 4.1) and 0.1 ml of hyaluronidase (340 units / ml) were added and incubated. (37 ° C, 20 minutes)
0.2 ml of Compound 48/80 (0.5 mg / ml) was added and further incubated at 37 ° C. for 20 minutes.
0.5 ml of hyaluronic acid (0.2 mg / ml) was added and further incubated at 37 ° C. for 40 minutes.
0.4 ml of 0.4N NaOH and boric acid solution (pH 9.1) equivalent amount mixed were put in,
After boiling for 3 minutes, it was cooled with water.
3 ml of P-DMAB was added, incubated at 37 ° C. for 40 minutes, and measured with a spectrophotometer. (585nm)

Each production example was dissolved in 0.1 M acetate buffer (pH 4.1) at a necessary concentration, subjected to the test, and the inhibition rate was calculated from the following formula.
A: With enzyme, without test product, B: With enzyme, with test product, C: Without enzyme, without test product, D: Without enzyme, with test product The results are shown in Table 1.

  When Production Examples 1 to 6 were used as external preparations for skin, they were effective for wrinkles and tarmi, and for contact dermatitis, psoriasis, pemphigus vulgaris, and the like.

Claims (1)

  A hyaluronidase inhibitor characterized by containing an extract from one or more plants selected from white birch, ishige, mulberry branch, and macomo.