CN101313913A - Hyaluronate lyase restrainer - Google Patents

Abstract

The invention relates to a hyaluronidase inhibitor containing new ingredients, and an anti-aging agent for skin and an anti-inflammatory agent which contain the hyaluronidase inhibitor. Active ingredients of the hyaluronidase inhibitor are phenylethanoid glycosides. The invention discloses that the hyaluronidase inhibitor has blocking activity against hyaluronidase, that is, the invention finds out that, in orobanchaceae, cistanche tubulosa has the highest content of phenylethanoid glycosides, and carries out purification for extracts of cistanche tubulosa for the first time to prepare phenylethanoid glycosides which are effective ingredients of the cistanche tubulosa. According to pharmacological experiments, the phenylethanoid glycosides have a notable effect on reinforcing memory and inhibiting the formation of a blood thrombus, etc.

Description

Hyaluronidase inhibitor

Technical field

The present invention relates to a kind of resisting age of skin agent and antiinflammatory that uses the Hyaluronidase inhibitor of novel ingredients and use this Hyaluronidase inhibitor, can be widely used in the medicine that comprises human mammal and use, skin preparations for extenal use, the food etc.

Background technology

Just know that since ancient times the herbal fresh stem of Orobanchaceae (Cistanche) is effective (referring to Patent Document 1) for the treatment of infertility, sexual impotence, constipation etc.And, can be used as the nutrition of blood and kidney by the prepared prepared product of fresh stem of these herbaceos perennials.These perennial parasitic herbaceous plant are extensively cultivated in northwest China, and some place is called them " desert Radix Ginseng ".Wherein holoparasite Cistanche Tubulosa Cistanche tubulosa (Schrenk.) Wight of Orobanchaceae is a kind of of cultivation output maximum in the Orobanchaceae (Cistanche).

Japan scholars etc. have carried out systematic research to chemical constituent and the pharmacologically active of Orobanchaceae (Cistanche) plant, found that the phenethyl alcohol glycoside compounds is the main active (for example, with reference to non-patent literature 1-3) of these herbaceos perennials.These active component can be used as effective anti-oxidants, metabolic improver, memory improving agent, libido promoter etc.Many researchers is furtherd investigate the medical science characteristic of multiple phenethyl alcohol glycoside compounds.

In addition, xerosis cutis, pachylosis, skin flexibility decrease, wrinkle increase these phenomenons and hyaluronic decomposition is closely related.Hyaluronic acid is a kind of mucopolysaccharide that extensively is present in the tissue such as vitreous body of skin, ligament, joint fluid and eyes; for example in skin or oral cavity; hyaluronic acid the transportation of the protection of cell, nutrition, organize moisture maintenance, organize aspects such as keeping of flexibility to play a significant role; and during as joint fluid, hyaluronic acid organizational structure and function keep and the aspects such as maintenance of lubricity play a significant role.In skin or oral cavity, joint, hyaluronic amount can or be in morbid state and reduces along with the old and feeble of human body in the organism, so by suppressing activity as the hyaluronidase of hyaluronic acid hydrolytic enzyme, can suppress hyaluronic decomposition, thereby help to keep hyaluronic content in the body.

In addition, because the variation of life style and the influence that chemical substance produced of variation to existing in the environment of various environmental conditions in recent years, simultaneously because the increase of pressure etc. in social life makes the crowd who the skin problem relevant with anaphylaxis such as urticaria, atopic dermatitis occur increase year by year.It is believed that for these skin problems, hyaluronic acid also is an important reasons.That is to say, if hyaluronidase is activated, hyaluronic acid is decomposed, and destroy intercellular substance in the conjunctive tissue wherein have hyaluronic acid etc., because the permeability of blood capillary etc. changes, the permeability of inflammatory cell is improved, causes acute edema thus, thus suppress hyaluronidase activity, prevent hyaluronic hydrolysis, keep the intercellular substance of conjunctive tissue to be very important.

Patent documentation 1: TOHKEMY 2004-250449 communique

Non-patent literature 1:Sato T, et al.Yakugaku Zasshi, 1985,105 (12) .1131

Non-patent literature 2:Jimenez C, et al Nat.Prod.Rep., 1994,11 (6): 591

Non-patent literature 3:Cometa F., et al.Fitoterapia, 1993,64 (3): 195

Summary of the invention

Purpose of design: under above-mentioned background, the inventor finds that the extract of Orobanchaceae plant, the phenethyl alcohol glycoside compounds that is particularly contained have hyaluronidase inhibitory activity in described extract, thereby finishes the present invention.That is to say, the object of the present invention is to provide a kind of hyaluronidase activity inhibitor of novelty and contain resisting age of skin agent, antiinflammatory of described hyaluronidase activity inhibitor etc.

Design: in order to address the above problem, the Hyaluronidase inhibitor described in the present invention is characterised in that it is an effective ingredient with the phenethyl alcohol glycoside compounds.In addition, as described phenethyl alcohol glycoside compounds, preferably contain at least a in echinacoside and the acteoside.Further, as described phenethyl alcohol glycoside compounds, preferably contain echinacoside and acteoside.Other Hyaluronidase inhibitor is characterised in that its extract with the Orobanchaceae plant is an effective ingredient among the present invention.In addition, as described extract, preferably wherein contain the phenethyl alcohol glycoside compounds as effective ingredient.Further, preferably, contain at least a as in the echinacoside of phenethyl alcohol glycoside compounds and the acteoside in the described extract.Preferably contain described echinacoside and described acteoside in the described extract.In addition, described Hyaluronidase inhibitor can be used as resisting age of skin agent or antiinflammatory.Further, the feedstuff that can use as medicine, skin preparations for extenal use, drinking food product, mammal that the mammal that comprises the people is used of described resisting age of skin agent or antiinflammatory.

Description of drawings

Fig. 1 is the figure of the hyaluronidase inhibitory activity of expression Cistanche Tubulosa extract.

Fig. 2 is the figure of the hyaluronidase inhibitory activity of expression echinacoside.

Fig. 3 is the figure of the hyaluronidase inhibitory activity of expression acteoside.

The specific embodiment

Below describe the present invention in detail.First invention is a kind of Hyaluronidase inhibitor, contains the phenethyl alcohol glycoside compounds in this Hyaluronidase inhibitor.

Herein, described " phenethyl alcohol glycoside compounds " is meant the represented chemical compound of following chemical formula (1).

Chemical formula (1)

Described phenethyl alcohol glycoside compounds is not limited especially; for example, can enumerate echinacoside; acteoside; 2 '-the acetyl group acteoside (2 '-acetylacteoside); campneoside I; campneoside II; Cistanche Tubulosa glycosides A (cistantubuloside A); Cistanche Tubulosa glycosides B1; Cistanche Tubulosa glycosides B2; Cistanche Tubulosa glycosides C1; Cistanche Tubulosa glycosides C2; crenatoside; remove coffee acyl acteoside (decaffeoylacteoside); Isoacteoside (isoacteoside); rhodioside (rhodioloside); syringalide A3 '-α-L-rhamnopyranoside and pipe flower glycosides A compositions such as (tubuloside A).In addition, these chemical compounds can only use a kind of, also can share more than 2 kinds.

In addition, preferably, contain at least a as in the echinacoside of described phenethyl alcohol glycoside compounds and the acteoside in the described Hyaluronidase inhibitor.This is because this Hyaluronidase inhibitor can have higher hyaluronic acid enzyme inhibition.

There is no particular limitation to the method that obtains the phenethyl alcohol glycoside compounds, can obtain by synthetic, also can obtain by extracting from plant.In addition, under situation about extracting from plant, there is no particular limitation to the plant as raw material, for example, can enumerate crude drug Radix Rehmanniae, labiate Herba Stachydis Japonicae, the fruit of Piperaceae Piper Fructus Canarii albi, the herbaceous plant of Orobanchaceae etc.In addition, can only use a kind of described plant to extract the phenethyl alcohol glycoside compounds, also can share more than 2 kinds plant and extract the phenethyl alcohol glycoside compounds.In these plants, most preferably use the plant of Orobanchaceae.This is because in the plant of Orobanchaceae; as the phenethyl alcohol glycoside compounds; except echinacoside and acteoside; also contain 2 '-the acetyl group acteoside (2 '-acetylacteoside); campneoside I; campneoside II; Cistanche Tubulosa glycosides A (cistantubuloside A); Cistanche Tubulosa glycosides B1; Cistanche Tubulosa glycosides B2; Cistanche Tubulosa glycosides C1; Cistanche Tubulosa glycosides C2; remove coffee acyl acteoside (decaffeoylacteoside); Isoacteoside (isoacteoside); rhodioside (rhodioloside); syringalide A 3 '-α-L-rhamnopyranoside and pipe flower glycosides A (tubuloside A) etc. are so have higher hyaluronidase inhibitory activity.Herein, there is no particular limitation to the Orobanchaceae plant as raw material, for example, can enumerate holoparasite Cistanche Tubulosa (Cistanche tubulosa), Saline Cistanche Herb (Cistanche salsa), Herba aeginetiae indicae (Aeginetia indica), Herba Boschniakiae Rossicae (Boschniakia rossica) etc., be limited to these plants but have more than.In addition, these plants can only be used a kind of, also can share more than 2 kinds.In these plants, preferably use holoparasite Cistanche Tubulosa (Cistanche tubulosa).

Composition in the above-mentioned phenethyl alcohol glycoside compounds and the relation between the described chemical formula (1) below are shown.But the structural formula of crenatoside is shown in following chemical formula (2).In described prepared product, only contain a small amount of or micro-echinacoside in mentioned component and other composition the acteoside basically.

Chemical formula (2)

Main component in the Hyaluronidase inhibitor

Chemical compound shown in the above-mentioned table 1 all can pass through high performance liquid chromatography (HPLC) and confirm.At this moment, the condition of high performance liquid chromatography is as follows.Immobile phase is a C18 alkyl silane bonded silica gel, and mobile phase is acetonitrile-0.05M phosphate aqueous solution (gradient is 4: 96 → 15: 85), and flow velocity is 1 ml/min, and the detection wavelength is 330nm.

Second kind of Hyaluronidase inhibitor of the present invention is characterised in that with the extract that extracted from the Orobanchaceae plant as effective ingredient.

Herein, there is no particular limitation to the plant as the Orobanchaceae of raw material, for example, can enumerate holoparasite Cistanche Tubulosa (Cistanche tubulosa), Saline Cistanche Herb (Cistanche salsa), Herba aeginetiae indicae (Aeginetia indica), Herba Boschniakiae Rossicae (Boschniakia rossica) etc., be limited to these plants but have more than.In addition, these plants can only be used a kind of, also can share more than 2 kinds.In addition, in these Orobanchaceae plants, preferably use holoparasite Cistanche Tubulosa (Cistanche tubulosa).

Further, preferably contain the phenethyl alcohol glycoside compounds in described " extract ".Herein; there is no particular limitation to described " phenethyl alcohol glycoside compounds ", can enumerate echinacoside; acteoside; 2 '-the acetyl group acteoside (2 '-acetylacteoside); campneoside I; campneoside II; Cistanche Tubulosa glycosides A (cistantubuloside A); Cistanche Tubulosa glycosides B1; Cistanche Tubulosa glycosides B2; Cistanche Tubulosa glycosides C1; Cistanche Tubulosa glycosides C2; crenatoside; remove coffee acyl acteoside (decaffeoylacteoside); Isoacteoside (isoacteoside); rhodioside (rhodioloside); syringalide A 3 '-α-L-rhamnopyranoside and pipe flower glycosides A compositions such as (tubuloside A).At this moment, a kind of mentioned component can be only contained in the described extract, also mixture of ingredients more than 2 kinds can be contained.Wherein, especially preferably contain at least a in echinacoside and the acteoside.

At this moment, there is no particular limitation to the content of echinacoside, with the Orobanchaceae plant (for example, the gross mass of extract holoparasite Cistanche Tubulosa (Cistanche tubulosa)) is during as 100 quality %, and the content of echinacoside is 10 quality %～70 quality %, be preferably 15 quality %～50 quality %, more preferably 20 quality %～40 quality %, 23 quality %～38 quality % more preferably.In addition, there is no particular limitation to the content of acteoside, with the gross mass of the extract of Orobanchaceae plant during as 100 quality %, the content of acteoside is 1 quality %～40 quality %, be preferably 5 quality %～30 quality %, more preferably 7 quality %～20 quality %, 8 quality %～10 quality % more preferably.

At this moment, there is no particular limitation to the position that is extracted of Orobanchaceae plant, and can use leaf, root, stem etc., as long as can reach desired effect.Preferably, from the stem of Orobanchaceae plant, particularly fresh stem, extract.

To from there is no particular limitation as the method that obtains extract the Orobanchaceae plant of raw material, for example, can enumerate solvent extraction method, super critical extraction etc.

In addition, when adopting solvent extraction method to extract, there is no particular limitation to employed solvent, but the preferred polar solvent that uses.In addition, there is no particular limitation to described polar solvent, for example, can enumerate water, methanol, ethanol, isopropyl alcohol, acetone, 1,3 butylene glycol, ethylene glycol, propylene glycol, glycerol, acetic acid, ethyl acetate, ether etc.These solvents can only use a kind of, also can share more than 2 kinds.Preferably, dilute subsequently, concentrate, processing such as drying, purification, because can obtain to contain the extract of the phenethyl alcohol glycoside compounds of high concentration by such processing.In addition, as purification process, for example, can enumerate charcoal treatment, resin absorption processing, ion exchange resin, liquid-methods such as liquid adverse current distribution.

Describe an embodiment of the concrete manufacture method of said extracted thing below in detail, but the manufacture method of extract is not limited to following embodiment.

The manufacture method of described extract is made of extraction and 2 stages of purification.In first stage, (for example, holoparasite Cistanche Tubulosa (Cistanche tubulosa)) stem (the fresh stem of this plant more preferably) is thinly sliced, and perhaps is ground into microgranule or Powdered with the Orobanchaceae plant.Next, with the thin slice that so obtains or microgranule or powder impregnation in lower aliphatic alcohols equal solvents such as water or methanol, ethanol or their mixed solution.At this moment, at room temperature extract.Then, filter this mixed solution, and concentrate this filtrate under reduced pressure or under the vacuum condition, obtain extract.Next, in second stage, after described extract heated, described extract is carried out purification in water by this extract being transferred in the adsorption column that is filled with D-101 type or AB-8 type macroporous adsorbent resin.At this moment, make the mixed liquor of water, methanol, ethanol, water and methanol or water and alcoholic acid mixed liquor etc., described adsorption column is carried out eluting as eluting solvent.During eluting, above-mentioned eluting solvent eluting can be come with the form of certain density solution, also above-mentioned eluting solvent eluting can be come according to Concentraton gradient.Collect eluent, after concentrating, adopt known drying means to carry out drying.After treating that the eluent drying is finished, obtain the said extracted thing.Contain the phenethyl alcohol glycoside compounds in the extract that so obtains.

Hyaluronidase inhibitor among the present invention can be as the raw material of various drinking food products.As described drinking food product, for example, can enumerate with edible oil (salad oil), snack categories (chewing gum, confection, caramel, chocolate, cookies, snacks, fruit jelly, chewable tablet, cookie etc.), Noodles (Fagopyrum esculentum Moench, Japanese noodle, hand-pulled noodles etc.), milk product (milk, popsicle, Yoghurt etc.), flavoring agent (miso, salty sauce etc.), soup class, beverage (fruit juice, coffee, black tea, tea, soda pop, sports drink etc.) is the normal food of representative or health food (tablet, capsule etc.), nutritional supplement (nutritious drink etc.).In these drinking food products, can suitably be furnished with cosmetic compositions of the present invention.

Kind according to these drinking food products, various compositions can be added therein, for example glucose can be used, fructose, sucrose, maltose, Sorbitol, stevioside (Stevioside), corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, the L-ascorbic acid, the dl-alpha-tocopherol, sodium erythorbate, glycerol, propylene glycol, fatty acid glyceride, polyglyceryl fatty acid ester, sucrose fatty acid ester, sorbitan aliphatic ester, methyl glycol fatty acid ester, arabic gum, chondrus ocellatus Holmes polysaccharide (carrageenan), casein, gelatin, pectin, agar, vitamin(e) B group, nicotiamide, calcium pantothenate, amino acids, the calcium salt class, pigment, spice, food materials such as preservative agent.In addition; also can add other antioxidant or adjuvant such as health food raw material in having the Hyaluronidase inhibitor of the present invention of keeping healthy functions, described adjuvant for example has antioxidant; reduced form ascorbic acid (vitamin C); vitamin E; reduced glutathion; tocotrienol; vitamin A derivative; lycopene; β-cryptoxanthin; astaxanthin; cryptoxanthin; rock algae Huang; uric acid; ubiquinone; coenzyme Q10; folic acid; Bulbus Allii extract; garlicin; sesamin; lignin; catechin; isoflavone; chalcone; the tannin class; flavonoid; coumarin; the isocoumarin class; Pericarpium Citri tangerinae extract; arbutin; tannin; anthocyanin; apple polyphenol; Semen Vitis viniferae extract; ellagic acid; kojic acid; Fructus Hippophae extract health food raw material; V. (vitamin) A; V.B1; V.B2; V.B6; V.B12; V.C; V.D; V.E; V.P; choline; nicotiamide; pantothenic acid; Calcium Folinate-SF; EPA; oligosaccharide; dietary fiber; Squalene; soybean lecithin; taurine; Dunaliella salina (Dunaliella); protein; octacosanol; DHA; Ovum Gallus domesticus Flavus lecithin; linoleic acid; lactoferrin; magnesium; zinc; chromium; selenium; potassium; heme; the Carnis ostreae extract; chitosan; the chitin oligosaccharide; collagen protein; chrondroitin; Radix Curcumae; Radix Glycyrrhizae; Fructus Lycii; Cortex cinnamomi japonici (Ramulus Cinnamomi); Fructus Crataegi; Rhizoma Zingiberis Recens; Ganoderma; a species of small clam living in fresh water shellfish extract; Trionyx sinensis Wiegmann; Radix Glycyrrhizae; Fructus Lycii; Cortex cinnamomi japonici (Ramulus Cinnamomi); Fructus Crataegi; Rhizoma Zingiberis Recens; Ganoderma; Herba Plantaginis; the Hemerocallis citrina Baroni chrysanthemum; Flos Matricariae chamomillae; Herba Taraxaci; Flos Hibisci Rosae-Sinensis; Mel; ボ one レ Application; Lac regis apis; the Lay lemon; lavandula angustifolia; Fructus Rosae Davuricae; Herba Rosmarini Officinalis; Salvia japonica Thunb.; bacillus bifidus; bacillus (faecalis); the sporozoite lactobacillus is arranged; Semen Tritici aestivi germ oil; Oleum Ricini; perilla oil; soybean oil; medium-chain fatty acid; Agaricus blazei Murrill (Agaricus); Folium Ginkgo extract; Radix Curcumae; chrondroitin; the brown rice germ extract; Ganoderma; Bulbus Allii Cepae; DHA; EPA; DPA; Folium hydrangeae strigosae; Cordyceps; Bulbus Allii; the Apis seed; Fructus Caricae; Folium camelliae assamicae; propolis; the farsighted person tree; ヤ Block シ ケ; Lac regis apis; Saw Palmetto P.E; hyaluronic acid; collagen protein; GABA; seal oil; shark cartilage; glucamine; lecithin; Phosphatidylserine; Radix Notoginseng; Folium Mori; soybean extract; the genus echinacea plant; Siberian ginseng (エ ゾ ウ コ ギ); Fructus Hordei Vulgaris extract; Folium olive; olive fruits; Largeleaf Gymnema (Gymnema); Flos Caryophylli Lagerstroemia indica L.; Salacia prinoides (Willd.) DC. platymiscium (Salacia); Garcinia plant (Garcinia); chitosan; Herba Hyperici perforati; Fructus Jujubae; Radix Ginseng; Herba Passiflorae Caeruleae; the green vegetable flower; Placenta Hominis; Semen Coicis; seed of Fructus Vitis viniferae; peanut skin; Pericarpium Citri tangerinae; Radix vernoniae asperae; Herba Silybi mariani; Laurel; Salvia japonica Thunb.; Herba Rosmarini Officinalis; Herba Apocyni veneti; black vinegar; go one ヤ one tea; the agate an ancient woman's ornament; Flos Carthami; Caulis et Folium Lini; oolong tea; the flower Herba Oxytropis Kansuensis; caffeine; capsaicin (capsaicin); xylooligosaccharide; glucamine; Semen Fagopyri Esculenti; citrus plant; dietary fiber; protein; dried plum; spirulina; the Fructus Hordei Vulgaris tender leaf; nucleic acid; yeast; Lentinus Edodes; plum pulp; aminoacid; deep-sea shark extract; noni (noni); Carnis ostreae; Trionyx sinensis Wiegmann; mushroom (シ ヤ Application ピ ニ オ Application); Herba Plantaginis; Fructus Pruni pseudocerasi (ア セ ロ ラ); Fructus Ananadis comosi; Fructus Musae; peach; Fructus Pruni; hami melon; Fructus Fragariae Ananssae; Fructus Rubi; Fructus Citri tangerinae; fucoidan; Phellinus igniarius (L. ex Fr.) Quel.; the crane certain kind of berries; chondroitin sulfate; zinc; ferrum; ceramide; silk peptide; glycine; nicotinic acid; Fructus Viticis; ceramide; the L-cysteine; the L-carnitine; the Radix seu Herba Tetrastigmatis Hypoglauci leaf; broomcorn millet; Herba Equiseti Arvinsis; biotin; Herba Centellae; Lonicera edulis Turcz.; pycnogenol (PYCNOGENOL); Petasites japonicus; Radix Et Rhizoma Rhei; Flos Caryophylli; Herba Rosmarini Officinalis; catechin; Folium camelliae assamicae; citric acid; beer yeast; メ リ ロ one ト; Semen sojae atricolor; Rhizoma Zingiberis Recens; Rhizoma Curcumae; Bacillus natto; Monas cuspurpureus Went; tocotrienol; lactoferrin; Cortex Cinnamomi; hull buckwheat; cocoa powder; the Fructus Citri grandis seed extract; purple common perilla seed; the Fructus Litchi seed extract; Radix Oenotherae erythrosepalae extract; the brown rice extract; alpha-lipoic acid; GABA; raw coffee bean extract; the Petasites japonicus extract; the Chinese gooseberry seed extract; Wenzhou Fructus Citri tangerinae extract; Rhizoma Zingiberis Recens extract; astaxanthin etc.

As concrete manufacture method, adopt spray drying or cryodesiccated method, can be directly with Hyaluronidase inhibitor of the present invention (if extract, then with this extract with dextrin) make powder, granule, tablet or solution, thereby easily they are included in the food (instant food etc.).In addition, as required, also Hyaluronidase inhibitor of the present invention or extract can be mixed with binding agents such as arabic gums, make Powdered or graininess, add to then in the solid type food.In addition, under the situation of extract, can directly this extract be added in the beverage, also can be in (for example) water, ethanol, glycerol or their mixture with this extract dispersing and dissolving, and then add in the beverage.

Hyaluronidase inhibitor of the present invention also can be used as the medicine raw material of (comprising pharmaceuticals and medicine part outer article).Can make described medicine by suitably adding Hyaluronidase inhibitor of the present invention in the raw material of using at medicine preparation.In addition, described medicine can be used for the mankind, also can be used for the mammal except the mankind.As can with the preparation raw material of Hyaluronidase inhibitor compatibility of the present invention, for example, can enumerate excipient (glucose, lactose, white sugar, sodium chloride, starch, calcium carbonate, Kaolin, crystalline cellulose, cocoa butter, plant fixed oil, Kaolin, Pulvis Talci etc.), binding agent (distilled water, normal saline, ethanol water, white sugar syrup, Glucose Liquid, starch fluid, gelatin solution, carboxymethyl cellulose, potassium phosphate, polyvinylpyrrolidone etc.), disintegrating agent (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, glyceryl monostearate, starch, lactose, the arabic gum powder, gelatin, ethanol etc.), disintegrate inhibitor (white sugar, stearic acid, cocoa butter, the hydrogenation wet goods), absorption enhancer (quaternary ammonium base, sodium lauryl sulfate etc.), adsorbent (glycerol, starch, lactose, Kaolin, Bentonite, silicic acid etc.), lubricant (purified talc, stearate, Polyethylene Glycol etc.) etc.

As the medication of Hyaluronidase inhibitor of the present invention, in general, can adopt form oral administrations such as tablet, pill, soft capsule, hard capsule, microgranule, powder, granule.In addition, as liquid preparation, water soluble preparation can oral administration, also can non-oral administration.As non-oral formulation and during administration, Hyaluronidase inhibitor of the present invention can be scattered in the suitable solvable reagent such as ethanol or water, use with dosage forms such as cataplasma, lotion, ointment, tincture, Emulsions then.In addition, the water soluble preparation of Hyaluronidase inhibitor of the present invention can directly use, and also can add dispersant, suspension, stabilizing agent etc. in this water soluble preparation, uses with dosage forms such as cataplasma, lotion, ointment, tincture, Emulsions then.

Dosage can change according to medication, the state of an illness, patient's age etc., but in general, the adult can be 5mg～200mg, child according to the effective ingredient that gave in 1 day according to the effective ingredient that gave in 1 day be that dosage about 0.5mg～100mg comes administration.

When Hyaluronidase inhibitor of the present invention used as medicine, the ratio of its adding can be come appropriate change according to dosage form.In general, oral administration or during by mucosa absorption and administration, the additional proportion of Hyaluronidase inhibitor can be about 0.01 weight %～10 weight %, but not during oral administration, the additional proportion of Hyaluronidase inhibitor can be for about 0.01 weight %～20 weight %.In addition, because dosage is according to the difference of various conditions and difference, so even the dosage that gives sometimes is also enough during less than above-mentioned dosage scope, dosage then needs above above-mentioned dosage range sometimes.Except containing above-mentioned Hyaluronidase inhibitor, also can contain known other chemical compound commonly used in the field of medicaments and can contain the needed chemical compound of molding when being suitable for peroral administration form in the medical composition.As such chemical compound, for example can enumerate lactose, starch, hydroxypropyl cellulose, Kaolin, Pulvis Talci, calcium carbonate etc.

In addition, in the feedstuff that mammal is used, also can contain Hyaluronidase inhibitor of the present invention.Can adopt the method identical, Hyaluronidase inhibitor of the present invention is included in the described animal feed with above-mentioned drinking food product.In addition, there is no particular limitation to the animal that can use described animal feed, and for example, this feedstuff can be used for companion animals such as livestock animals such as cattle, pig, Canis familiaris L., cat, hamster (as house pet and domesticated animal) etc.In addition, for example,, can use flour, edible meat etc. as the feedstuff of described companion animals.At this moment, as described flour, wheat flour, rice flour, rye meal, oatmeal, barnyard grass flour, millet powder, Semen Maydis powder, Semen sojae atricolor powder etc. are for example arranged, these flour also can share more than 2 kinds.By using described flour, nutrients such as carbohydrate are provided for described companion animals.In the described flour, most preferably use wheat flour.As described wheat flour, can be separately or appropriate combination use hard flour, middle power powder, weak flour, described in addition wheat flour also can share with other flour.In addition, in order to regulate the elasticity of the animal feed after heat treated, wheat flour and Testa Tritici, soybean protein etc. are used in combination.And the network structure of the wheat bran that is contained in the wheat flour can form expanded organizational structure after heat treated, help to improve mouthfeel.

There is no particular limitation to employed edible meat among the present invention, can use Carnis Gallus domesticus, Carnis Sus domestica, beef, Carnis caprae seu ovis, Carnis Naemorhedi, Carnis Leporis, turkey meat, Equus caballus (L.) etc., considers the best Carnis Gallus domesticus that uses from taste.Can after adopting method commonly used to slaughter livestock animal and dissection, obtain described edible meat.And the product generation deterioration of middle moisture product or low moisture state mainly is that the oxidation owing to fat causes, thus employed edible meat preferably fat content few or removed fatty lean meat.In addition, in feedstuff, contain edible meat, not only can provide good animal protein, but also can improve the hobby of described companion animals to animal.

Prepare the feedstuff of described companion animals though can adopt the whole bag of tricks, but preferable methods for example has such method: preparation contains the raw mix that mixes and stirs (hereinafter being called " dough/pasta ") of flour (being preferably wheat flour) and edible meat, carries out heat treated again after to be formed.There is no particular limitation to the composition of flour in the described dough/pasta and edible meat, in general, be mixed with by about 5%～60%, be preferably the flour about 10%～50%, about 5%～80%, be preferably the dough/pasta that the moisture of edible meat about 20%～50% and necessary amounts is formed.And under the situation of using the Aw regulator, the addition of this Aw regulator is about 5%～30%, is preferably about 10%～20%.In addition, the consumption of water can suitably be regulated according to the consumption of the mixing stirring of described dough/pasta, the degree that is shaped and flour, edible meat, Aw regulator etc.There is no particular limitation to the compound method of described dough/pasta, preferably, at first with cutmixer, chipper etc. described edible meat made meat stuffing.At this moment, in order in making the edible meat of meat stuffing, fully to contain bubble, preferably described edible meat is frittered.Then, in the described edible meat of making meat stuffing, add flour, water and can also add Aw regulator etc. as required, fully stir to make and wherein contain bubble, thereby be mixed with the dough/pasta that contains bubble.When the preparation dough/pasta, also can add foaming agent, particularly be used as under the situation of described flour at other flour except wheat flour, preferably use foaming agent.By adding foaming agent, can make and contain micro-bubble in the dough/pasta equably.As described foaming agent, can use various foaming agent, but consider from the aspects such as stability of bubble, preferably use soybean protein class foaming agent and/or zymolytic soybean protein class foaming agent.

After making the dough/pasta molding and process heat treated that so makes, obtain the feedstuff that described companion animals of the present invention is used.When the molding of dough/pasta, according to described companion animals in the easy degree in when food, raise the easy degree of people's operation etc., described companion animals of the present invention can be made suitable shape with feedstuff, for example described lamellar, bar-shaped, disk shape, ring-type, heart type etc. of being shaped as.In addition, can with by the prepared dough/pasta of identical matching component with the pigment dyeing of various different tones or add vegetable or fruit etc., thereby make multiple dough/pasta, these dough/pastas are combined into multiwalled shape or concentrically ringed shape, thereby molding with different outward appearances.There is no particular limitation to the heater means of the dough/pasta after the molding, and for example, described heater means can be enumerated baking oven for heating, microwave heating etc.These heating means all are known, can carry out heat treated according to method commonly used.Be generally about 20%～40% through the moisture in the feedstuff after the heat treated.Through after the described heat treated, because the evaporation of moisture and the expansion of bubble, described dough/pasta is expanded, in addition owing to be to make water evaporates at short notice, so Aw reduce, thereby improved keeping quality.In addition, as under the situation of described flour, by heat treated, the network structure of the wheat bran that is contained in the wheat flour is fixed, thereby improves mouthfeel at wheat flour.And, under the situation that adopts baking oven for heating, have and in described feedstuff, produce the unique tone (Vulpes color) or the advantage of fragrance, and under the situation that adopts microwave heating, owing to be to begin heating in the inside of dough/pasta, so can make the dough/pasta uniform expansion, have the such advantage of feedstuff that can obtain containing even bubble.When carrying out described heat treated, preferably, Aw in the resulting feedstuff is adjusted in 0.6～0.9 the scope.As mentioned above, by Aw being adjusted in this scope, can significantly improve the keeping quality of feedstuff.

The feedstuff of the described companion animals that so makes is the feedstuff with character of bread, because this feedstuff has softish mouthfeel and suitable flexibility and elasticity, so be suitable as the feedstuff, dessert etc. of the weak companion animals of tooth abilities such as pup, aged dog or cat.Above-mentioned animal feed is contained in the packing container with suitable amount, makes product after the sealing.As described packing container, preferably use oxygen impermeable packaging material.As the shape of packing, can enumerate vacuum packaging, be filled with the packing of active carbon etc., but preferably be filled with the packing of deoxidant (for example, エ one ジ レ ス TM etc.) and active carbon simultaneously.Adopt relevant packaged form, during preservation can prevent the deterioration that causes owing to oxygen and the breeding of microorganism.

Hyaluronidase inhibitor of the present invention also can be used as skin preparations for extenal use (comprising cosmetics, pharmaceuticals and medicine part outer article), expects that this skin preparations for extenal use has the hyaluronic acid enzyme inhibition.In addition, described skin preparations for extenal use can be used for the mankind, also can be used for other mammal except the mankind.As the form that the skin preparations for extenal use of Hyaluronidase inhibitor of the present invention can compatibility be arranged, for example, can enumerate emulsion, soap, the agent of washing one's face, bathing agent, protective skin cream, emulsion, astringent, floral water, shave with emulsifiable paste, shave with washing liquid, greasepaint, Sun block astringent, muffin, foundation cream, perfume, facial film (pack), nail polish paste, nial polish (enamel), nial polish washing liquid, brow, kermes, eye cream, eye shadow, mascara, informer, lipstick, lip pomade, shampoo, hair conditioner, hair dye, dispersion liquid, detergent.In addition, as the form that the pharmaceuticals or the medicine part outer article of Hyaluronidase inhibitor of the present invention can compatibility be arranged, can enumerate ointment, Emulsion, external liquid etc.

In the skin preparations for extenal use of above-mentioned form, except containing Hyaluronidase inhibitor of the present invention, in not damaging the inhibiting scope of hyaluronidase, also can add following substances, described material comprises oils composition, higher alcohol, fatty acid, UV absorbent, powder, pigment, surfactant, polyhydric alcohol, saccharide, macromolecule, biological active substances, solvent, antioxidant, spice, antiseptic etc. for joining the composition in the skin preparations for extenal use such as cosmetics, medicine part outer article.Though below enumerate out these examples of substances, the present invention has more than and is limited to these examples.

(1) the oils composition of the example esters of oils composition has for example: three 2 ethyl hexanoic acid glyceride, 2 ethyl hexanoic acid hexadecanol ester, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearte, octyl palmitate, the different hexadecanol ester of isostearic acid, butyl stearate, butyl myristate, Ethyl linoleate, the linoleic acid isopropyl ester, ethyl oleate, the different hexadecanol ester of myristic acid, myristic acid i-octadecanol ester, Palmic acid i-octadecanol ester, octyl dodecyl myristate, the different hexadecanol ester of isostearic acid, ethyl sebacate, diisopropyl adipate, different 10 alcohol esters of neopentanoic acid, three (octyl group capric acid) glyceride, three 2 ethyl hexanoic acid trihydroxy methyl propyl ester, three isostearic acid trihydroxy methyl propyl ester, four 2 ethyl hexanoic acid pentaerythritol esters, sad hexadecanol ester, lauric acid ester in the last of the ten Heavenly stems, lauric acid hexyl ester, myristic acid ester in the last of the ten Heavenly stems, myristic acid Semen Myristicae alcohol ester, myristic acid hexadecanol ester, stearic acid octadecanol ester, decyl oleate, ricinoleic acid hexadecanol ester, lauric acid i-octadecanol ester, myristic acid different three the last of the ten Heavenly stems ester, the different hexadecanol ester of myristic acid, myristic acid i-octadecanol ester, the different hexadecanol ester of Palmic acid, Palmic acid i-octadecanol ester, octyl stearate, the different hexadecanol ester of stearic acid, Ceraphyl 140A, oleic acid octyl dodecanol ester, linoleic acid octyl dodecanol ester, the isostearic acid isopropyl ester, 2 ethyl hexanoic acid octadecanol hexadecanol ester, 2 ethyl hexanoic acid octadecanol ester, the own ester of isostearic acid, Ethylene glycol dicaprylate, two oleic acid glycol esters, the didecyl acid propylene glycol ester, two (octyl group capric acid) propylene glycol ester, two sad propylene glycol esters, two capric acid DOPCP, two sad DOPCP, tricaprylin, three (undecanoic acid) glyceride, three different tripalmitins, three glyceryl isostearates, neopentanoic acid octyl dodecanol ester, sad i-octadecanol ester, different octyl pelargonate, new decanedioic acid hexyl lauryl alcohol ester, new decanedioic acid octyl dodecanol ester, the different hexadecanol ester of isostearic acid, isostearic acid i-octadecanol ester, isostearic acid octyl dodecanol ester, polyglycerol acrylate, polyglyceryl-isostearate, dipropyl carbonate, dialkyl carbonate (C ₁₂-C ₁₈) ester; citric acid three different hexadecanol esters; citric acid three different 10 alcohol esters; citric acid three different monooctyl esters; Lauryl lactate; Tetradecyl lactate; cetyl lactate; lactic acid octyl dodecanol ester; triethyl citrate; acetyl triethyl citrate; acetyl tributyl citrate; trioctyl lemon acid; malic acid two i-octadecanol esters; hydroxy stearic acid 2-Octyl Nitrite; succinic acid two 2-Octyl Nitrites; diisobutyl adipate; Dermol DIPS; di-n-octyl sebacate; cholesteryl stearate; the isostearic acid cholesterol ester; the hydroxy stearic acid cholesterol ester; Cholesterol 3.beta.-oleate; oleic acid dihydrocholesterol ester; the isostearic acid plant sterol ester; the oleic acid plant sterol ester; the different hexadecanol ester of 12-stearyl hydroxy stearic acid; 12-stearyl hydroxy stearic acid octadecanol ester; 12-stearyl hydroxy stearic acid i-octadecanol ester etc.The oils composition of hydro carbons has for example: Squalene, liquid paraffin, alpha-olefin low polymers, different paraffin, ceresin, paraffin, liquid different paraffin, polybutene, microwax, vaseline etc.Vegetable and animals oils and fixed oil thereof and natural wax have for example: Adeps Bovis seu Bubali, sclerosis Adeps Bovis seu Bubali, leaf fat, sclerosis leaf fat, house oil, sclerosis house oil, ermine oil, オ レ Application ジ ラ Off イ one (a kind of fish of sour jujube Channa argus section) oil, fish oil, sclerosis fish oil, egg yolk wet goods animal oil and fixed oil thereof; American Avocado Tree oil, almond oil, olive oil, cocoa butter, the Fructus actinidiae chinensis seed oil, almond oil, Aloe arborescens var. natalensis oil, Oleum sesami, Semen Tritici aestivi germ oil, rice germ oil, profound Miyou, safflower oil, shea butter, Oleum Glycines, Radix Oenotherae erythrosepalae oil, perilla oil, Oleum Camelliae, Camellia oil, Semen Maydis oil, Oleum Brassicae campestris, hardened rapeseed oil, palm kernel oil, the sclerosis palm kernel oil, Oleum Arachidis hypogaeae semen, sclerosis Oleum Arachidis hypogaeae semen, Oleum Ricini, hardened castor oil, Oleum helianthi, Oleum Vitis viniferae, Jojoba oil, the sclerosis Jojoba oil, macadimia nut oil, メ De ホ one system oil, Oleum Gossypii semen, the sclerosis Oleum Gossypii semen, Oleum Cocois, sclerosis Cortex cocois radicis wet goods vegetable oil and fixed oil thereof; Waxes such as Cera Flava, high acid value Cera Flava, lanoline, reduction lanoline, sclerosis lanoline, aqueous lanoline, Brazil wax, montan wax.The oils composition of type siloxane has for example: dimethyl polysiloxane, methyl phenyl silicone, the methyl cyclopolysiloxane, the prestox polysiloxanes, the decamethyl polysiloxanes, ten dimethyl polysiloxanes, methylhydrogenpolysi,oxane, polyether-modified organopolysiloxane, dimethyl siloxane methyl cetyl hydroxyl silicone copolymers, dimethyl siloxane methyl octadecyl hydroxyl silicone copolymers, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino modified silicone oil, amino modified organopolysiloxane, dimethyl silica alkanol (Dimethiconol), silicone adhesive, the acrylic silicone, trimethylsiloxy silicic acid, silicone RTV glue etc.The oils composition of fluorine class has for example: PFPE, fluorine modification organopolysiloxane, fluoridize the poly-hydroxy alkylidene of Colophonium, perfluorocarbon, fluorinated alcohols, fluoro-alkyl modification organopolysiloxane etc. altogether.

(2) example of higher alcohol has: lauryl alcohol, myristyl alcohol, spermol, octadecanol, isooctadecane alcohol, oleyl alcohol, docosanol, 2-Ethylhexyl Alcohol, hexadecanol, octyldodecanol etc.

(3) example of fatty acid has: sad, capric acid, 9-undecylenic acid, lauric acid, myristic acid, Palmic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid, sinapic acid, 2 ethyl hexanoic acid etc.

(4) example of UV absorbent has: para-aminobenzoic acid; the para-aminobenzoic acid pentyl ester; para-aminobenzoic acid ethyl dihydroxy propyl ester; para-aminobenzoic acid glyceride; the para-aminobenzoic acid ethyl ester; the para-aminobenzoic acid monooctyl ester; para-aminobenzoic acid octyl group dimethyl ester; glycol salicylate; ethylhexyl salicylate; trolamine salicylate; phenyl salicytate; salicylic acid butyl phenyl ester; benzyl salicylate; salicylic acid is with ester in the Meng; benzyl cinnamate; the p-methoxycinnamate monooctyl ester; p-methoxycinnamate 2-Octyl Nitrite; di-p-methoxy cinnamic acid single 2-ethyl caproin; the p-methoxycinnamate isopropyl ester; the p-methoxycinnamate diethanolamine; diisopropyl diisopropyl cinnamic acid ester mixture; urocanic acid; the urocanic acid ethyl ester; hydroxyl methoxybenzene ketone; hydroxyl methoxybenzene ketone sulfonic acid and salt thereof; dihydroxy methoxybenzene ketone; dihydroxy methoxybenzene ketone sodium sulfonate; the dihydroxy benzenes ketone; dihydroxy dimethoxy benzophenone; hydroxyl octyloxy benzophenone; the tetrahydroxy benzene ketone; PAROSOL 1789; 2; 4; 6-triphen amido-right-(2-ethylhexyl-1-carboxyl)-1; 3; the 5-triazine; 2-(2-hydroxy-5-methyl base phenyl) benzotriazole; neighbour-Methyl anthranilate; 2-amino-3; 3-diphenylacrylate 2-Octyl Nitrite; phenyl benzimidazole sulphuric acid; 3-(4-methylbenzene methylene) Camphora; isopropyl diphenyl formoxyl methane; 4-(3; 4-dimethoxy benzene methylene)-2; 5-dioxo-1-imidazolidinyl propanoic acid 2-Octyl Nitrite etc., and the polymeric derivative of these materials or silane derivative etc.

(5) example of powder, pigment has: pigments such as No. 104, redness, No. 201, redness, No. 4, yellow, No. 1, blueness, No. 401, black; Precipitation color elements such as yellow No. 4 AL lakes, yellow No. 203 BA lakes; Macromolecules such as nylon powder, silk powder, urethanes powder, polytetrafluoroethylene (registered trade mark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, starch, the spherical elastomer powder of silicone, polyethylene powder; Yellow iron oxide, red iron oxide, black iron oxide, chromium oxide, white carbon black, ultramarine, colored pigment such as dark blue; Chinese whites such as zinc oxide, titanium oxide, cerium oxide; Extender pigments such as Pulvis Talci, Muscovitum, cerussite, Kaolin, tabular barium sulfate; Pearlescent pigments such as mica titanium; Slaines such as barium sulfate, calcium carbonate, magnesium carbonate, aluminium silicate, magnesium silicate; Inorganic powder such as silicon, aluminium oxide; Metallic soaps such as aluminium stearate, magnesium stearate, Hexadecanoic acid, zinc salt, Grillocin P 176, magnesium myristate, Dodecanoic acid, zinc salt, Zinc Undecylenate; Bentonite, Montmorillonitum, boron nitride etc.There is no particular limitation to the shape of these powder (spherical, bar-shaped, needle-like, tabular, indefinite form, flakey, spindle shape etc.) and particle diameter.In addition; also can adopt known surface treatment method (as; fluorine compounds processing method, silicone-treated method, silicone treated method, the facture of dangling, silane coupling agent facture, titanium couplant facture, oil preparation facture, N-acyl group resin treatment method, polyacrylic acid facture, metallic soaps facture, aminoacid facture, lecithin facture, inorganic compound facture, plasma processing method, mechanochemistry facture etc.) these powder are carried out surface treatment in advance.

(6) the example anionic surfactant of surfactant can have been enumerated: fatty acid soaps; α-acyl group sulfonate; alkylsulfonate; alkylallyl sulfonate; alkylnaphthalene sulfonate; alkyl sulfate; the POE alkyl ether sulfate; alkyl amino sulfate; alkylphosphonic; the POE alkylphosphonic; alkyl amino phosphate; the alkanoyl alkyltaurate; N-acyl amino hydrochlorate; the POE alkyl ether carboxy acid salt; alkyl sulfo succinate; alkyl sulfoethanoic acid. sodium; acyl group hydrolytic collagen peptide salt; perfluoralkyl phosphate etc.Cationic surface active agent can have been enumerated: alkyl trimethyl ammonium chloride; the stearyl trimethyl ammonium chloride; the stearyl trimethylammonium bromide; octadecanoyl palmitoyl trimethyl ammonium chloride; distearyl acyl group alkyl dimethyl ammonium chloride; the stearyl dimethyl benzyl ammonium chloride; 22 carbonic acyl radical trimethylammonium bromides; the chlorination bromo geramine; behenic acid aminopropyl dimethyl hydroxyl propyl ester ammonium chloride; stearic acid diethyl amino yl acetamide; stearic acid dimethylamino propionic acid amide.; lanolin derivative quaternary ammonium salt etc.Amphoteric surfactant can have been enumerated: carboxybetaine type, amido betaine type, sulfobetaines type, hydroxyl sulfo betaine type, amidosulfobetaines type, phosphoric acid betaine type, aminocarboxylate type, imidazolidine derivatives type, amido amine type etc.Nonionic surfactant can have been enumerated: methyl glycol fatty acid ester, fatty acid glyceride, polyglyceryl fatty acid ester, Sorbitol fatty acid ester, POE fatty acid esters of sorbitan, POE Sorbitol fatty acid ester, POE fatty acid glyceride, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POEPOP copolymer, POEPOP alkyl ether, polyether modified siloxane lauric acid alkylolamides, alkyl amine oxide, hydrogenated soy phosphatidyl choline etc.Natural surfactant can have been enumerated: lecithin, saponin, sugar surfactants etc.

(7) example of polyhydric alcohol, sugar can have been enumerated: ethylene glycol, diethylene glycol, Polyethylene Glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerol, two glycerol, polyglycereol, 3-methyl isophthalic acid, 3-butanediol, 1,3 butylene glycol, sorbitol, mannitol, melitriose, erythritol, glucose, sucrose, fructose, xylose, lactose, maltose, maltose alcohol, trehalose, alkylation trehalose, mixing opposite sex sugar, sulphation trehalose, amylopectin etc.Also can use their chemical modification object etc. in addition.

(8) high molecular example has: acrylate/methacrylate copolymer (プ ラ ス サ イ ズ, mutual induction KCC makes), vinylacetate/.beta.-methylacrylic acid copolymer (レ ジ Application 28-1310, Niigata Seimitsu Co., Ltd. makes), vinylacetate/.beta.-methylacrylic acid/vinyl neodecanoate copolymer (28-2930, Niigata Seimitsu Co., Ltd. makes), methyl vinyl ether maleic acid half ester (ガ Application ト レ Star Star ES, ISP Co., Ltd. makes), tert-butyl acrylate/ethyl acrylate/methacrylic acid copolymer (Le PVC マ one, BASF Co., Ltd. makes), vinyl pyrrolidone/vinyl acetate/vinyl propionate ester copolymer (Le PVC ス コ one Le VAP, BASF Co., Ltd. makes), vinyl-acetic ester/.beta.-methylacrylic acid copolymer (Le PVC セ Star ト CA, BASF Co., Ltd. makes), vinyl-acetic ester/.beta.-methylacrylic acid/vinylpyrrolidone copolymer (Le PVC セ Star ト CAP, BASF Co., Ltd. makes), vinyl pyrrolidone/acrylate copolymer (Le PVC Off レ Star Network ス, BASF Co., Ltd. makes), acrylate/acrylamide copolymer (ウ Le ト ラ ホ one Le De, BASF Co., Ltd. makes), vinyl-acetic ester/maleic acid butyl ester/isobornyl acrylate copolymer (ア De バ Application テ one ジ, ISP Co., Ltd. makes), CVP Carbopol ETD2050 (カ one ボ Port one Le, BFGoodrich company makes), acrylic acid methacrylic acid alkyl copolymer (ペ ミ ユ レ Application, BFGoodrich company makes) etc. the anionic macromolecular compound; Perhaps the acetic acid amphoteric compound of methacrylic acid dialkyl amido polymerizable methacrylate thing (ユ カ Off オ one マ one, Mitsubishi chemical Co., Ltd make), acrylic acid octyl acrylamide/acrylic acid hydroxy propyl ester/metering system acid butyl amino ethyl ester copolymer amphoteric polymer compounds such as (AMPHOMER, Niigata Seimitsu Co., Ltd. make); The season carbide of vinyl pyrrolidone/dimethylaminoethyl methacrylate (GAFQUAT, ISP company make), methyl ethylene imidazolitm chloride/vinylpyrrolidone copolymer cationic high-molecular chemical compounds such as (Le PVC コ one ト, BASF Co., Ltd. make); Polyvinylpyrrolidone (Le PVC ス コ one Le K, BASF Co., Ltd. makes), vinylpyrrolidone/vinyl acetate base ester copolymer (Le PVC ス コ one Le VA, BASF Co., Ltd. makes), vinylpyrrolidone/dimethylaminoethyl methacrylate copolymer (コ Port リ マ 1, ISP company makes), caprolactam/vinylpyrrolidone/dimethylaminoethyl methacrylate copolymer (コ Port リ マ one VC713, ISP company makes) etc. non-ionic macromolecule compound etc.In addition, followingly derive from natural macromolecular compound and also be suitable for: cellulose and derivant thereof, keratin and collagen protein or their derivant, calcium alginate, pulullan polysaccharide, agar, gelatin, tamarind seed polysaccharide, xanthan gum, chondrus ocellatus Holmes polysaccharide, hyper-methoxy pectin, hypo-methoxy pectin, guar gum, arabic gum, crystalline cellulose, arabinogalactan, karaya, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, glucosan etc.

(9) example of physiologically active ingredient in the situation on being coated on skin, can be enumerated the material that those can bring certain physiologically active to skin as physiologically active ingredient.For example, can enumerate whitening composition, immunity effect agent, age resister, ultraviolet screening agent, weight reducing agent, stretching agent, antioxidant, increase a mao agent, educate a mao agent, wetting agent, blood circulation promoting agent, antibacterial, antibacterial, desiccant, creeping chill agent, temperature sense agent, vitamins, aminoacid, wound healing promoter, stimulate demulcent, analgesics, cell vitalizing agent, enzyme component etc.As with the example of the appropriate ingredients of these physiologically active ingredient compatibilities, for example have: return positive careless extract, the American Avocado Tree extract, the Radix Hydrangeae Strigosae extract, the Althaea rosea (L.) Cavan. extract, the arnica montana extract, Aloe extract, Fructus Pruni extract, the almond extract, Semen Ginkgo extrac, Fructus Foeniculi extract, Radix Curcumae extract, oolong tea extract, the Fructus rosae multiflorae extract, the SONGGUOJU leaf extract, Radix Scutellariae extract, the Cortex Phellodendri extract, Rhizoma Coptidis extract, Fructus Hordei Vulgaris extract, the Herba Hyperici Erecti extract, the Herba lamii barbati extract, bean is broken the dish extract off with the fingers and thumb, the Fructus Citri tangerinae extract, the dry thing of sea water, Sargassum extract, elastin hydrolysis, hydrolysis Semen Tritici aestivi powder, hydrolyzed-silk, camomile extract, Radix Dauci Sativae extract, the Herba Artemisiae Scopariae extract, Radix Glycyrrhizae extract, Hibiscus rosa-sinensis's extract, Pyracantha extract, the quinine extract, Fructus Cucumidis sativi extract, guanosine, Fructus Gardeniae extract, the Sasa veitchii(Carr.)Rehd. extract, Radix Sophorae Flavescentis extract, Juglans regia L. extract, grapefruit extract, the silk ball rattan extract, spirulina extract, the Mulberry extract, Radix Gentianae extract, black tea extract, yeast extract, the Fructus Arctii extract, profound rice fermented product extract, rice germ oil, the Symphytum officinale extract, ossein, Pericarpium Citri tangerinae extract, Herba Asari extract, Radix Bupleuri extract, the umbilical cord extracting solution, sage extract, the Saponaria officinalis extract, little bamboo extract, Fructus Crataegi extract, Fructus Zanthoxyli extract, Lentinus Edodes extract, Radix Rehmanniae extract, the Rhizoma Seu Herba Bergeniae extract, Folium perillae extract, the tilia extract, Ramulus et Folium Spiraeae Salicifoliae extract, Radix Paeoniae Alba extract, the Rhizoma Acori Graminei extract, the Betula platyphylla Suk. extract, Herba Equiseti Arvinsis extract, Caulis Hederae Sinensis extract, the Mespilus germinica Linn extract, Ramulus Sambuci Williamsii extract, the yarrow extract, the West Folium Menthae extract, sage extract, the Radix Malvae sylvestris extract, Rhizoma Chuanxiong extract, Herba Swertiae bimaculatae extract, soybean extract, Fructus Jujubae extract, Thymi Serpylli Herba extract, tea extract, Flos Caryophylli extract, the Rhizoma Imperatae extract, Pericarpium Citri Reticulatae extract, Radix Angelicae Sinensis extract, Calendula officinalis extract, Semen Persicae extract, the juniper extract, Herba Houttuyniae extract, the Fructus Lycopersici esculenti extract, natto extract, Radix Ginseng extract, the Bulbus Allii extract, the Flos rosae multiflorae extract, Hibiscus rosa-sinensis's extract, the Radix Ophiopogonis extract, parsley extract, Mel, the Radix Hamamelidis Mollis extract, summer Flos Chrysanthemi extract, Herba Rabdosiae glaucocalycis extract, bisabolol, the Folium Eriobotryae extract, Herba Taraxaci extract, the Petasites japonicus extract, Poria extract, the butchers broom extract, Fructus Vitis viniferae extract, propolis, Fructus Luffae extract, Flos Carthami extract, the Mentha arvensis L. syn.M.haplocalyxBrig extract, the bodhi tree extract, peony extract, Flos lupuli (Flos Humuli Lupuli) extract, the pine extract, the Aesculus hippocastanum L. extract, the water plantain extract, the Fructus Sapindi Mukouossi extract, the Herba Melissae officinalis extract, the peach extract, the Centaurea cyanus extract, eucalyptus extracts, Herba Saxifragae extract, Semen Coicis extract, biennial wormwood extract, Herba Lysimachiae foenum-graeci extract, Fructus Mali pumilae extract, Caulis et Folium Lactucae sativae extract, citron extract, the Herba Astragali Melilotoidis (Herba Astragali Sinici) extract, the Flos Rosae Multiflorae extract, Herba Rosmarini Officinalis extract, the Roman Herba Cancriniae Discoideae extract, Lac regis apis extract etc.Can also enumerate in addition: biopolymer such as DNA (deoxyribonucleic acid), mucopolysaccharide, hyaluronate sodium, sodium chondroitin sulfate, ossein, elastin laminin, chitin, chitosan, hydrolysed egg shell membrane; The compositions of preserving moisture such as aminoacid, range of hydrolysed peptides, sodium lactate, carbamide, pyrrolidone sodium carboxylate, betanin, milk surum, trimethyl glycine; Oiliness compositions such as neural lipid, ceramide, phytosphingosine, cholesterol, cholesterol derivative, phospholipid; Immunity effect agent such as episilon amino caproic acid, glycyrrhizic acid, β-enoxolone, lysozyme chloride, guaiazulene, ヒ De ロ コ one Le チ ゾ Application; Vitaminss such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, vitamin C ester; Allantoin, DIEDI, 4-amino methyl cyclohexylenedinitrilotetraacetic acid isoreactivity composition; Antioxidants such as tocopherol, carotenoid, flavone, tannin, lignin, saponin; Cell vitalizing agent such as alpha-hydroxy acid, β-hydroxy acid; Blood circulation accelerant such as gamma oryzanol, vitamin e derivative; The agent of wound healing such as retinol, retinol derivatives; Whitening agents such as albumin, kojic acid, intacellin, sulfur, ellagic acid, linoleic acid, tranamic acid, glutathion; Cephalanthin; Radix Glycyrrhizae extract; capsaicin; sabinol; iodate Bulbus Allii extract; pyridoxine hydrochloride; the DL-alpha-tocopherol; acetic acid DL-alpha-tocopherol; nicotinic acid; nicotinic acid derivates; calcium pantothenate; D-panthenol; acetyl group pantoyl ethylether; biotin; allantoin; isopropyl methyl phenol; estradiol; ethinyl estradiol; carpronium Chloride; benzalkonium chloride; diphhydramine hydrochloride; カ Na one Le; Camphora; salicylic acid; n-nonanoic acid vanilla amide; nonane acid vanilla amide; ピ ロ Network ト Application オ ラ ミ Application; pentadecanoic acid glyceride; the L-menthol; single nitroguaiacol; resorcinol; γ-An Jidingsuan; benzethonium chloride; mexiletine hydrochloride; auximone; estrogen; Mylabris tincture; cyclosporin; Zinc Pyrithione; ヒ De ロ コ one Le チ ゾ Application; minoxidil; monostearate polyoxyethylene sorbitan; Oleum menthae; サ サ ニ シ キ rice extracts etc. is educated mao agent etc.

(10) examples of antioxidants can be enumerated: sodium sulfite, sodium sulfite, arabo-ascorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, the tolyl biguanide, nordihydroguaiaretic acid, MEHQ, butylated hydroxyanisole (BHA), dibenzylatiooluene, ascorbyl stearate, ascorbic palmitate, gallateoctylester, propyl gallate, carotenoid, flavone, tannin, lignin, saponin, Fructus Mali pumilae extract and Flos Caryophylli extract etc. have the plant extract of antioxidant effect etc.

(11) example of solvent can be enumerated: purified water, ethanol, lower alcohol, ethers, LPG, fluorocarbon, N-Methyl pyrrolidone, fluorine substituted alcohols, have volatile linear siloxanes, secondary freon etc.

Embodiment 1: though embodiments of the invention are described, the present invention is not limited to these embodiment.

Embodiment: the preparation embodiment 1 of Hyaluronidase inhibitor of the present invention: the preparation of Orobanchaceae holoparasite Cistanche Tubulosa (Cistanche tubulosa) (hereafter is " Cistanche Tubulosa ") extract

Extraction process: the thin slice 10kg of the fresh stem of Cistanche Tubulosa be impregnated in the water of 8 times of amounts of this thin slice.Thin slice flooded 1 hour in water after, water decocted 2 hours.Mixture after so decocting is filtered, obtain first filtrate.Next, after 40% ethanol of 4 times of amounts of residue and this residue of the mixture after decocting mixed, decocted 4 hours, the mixture after the decoction is filtered, obtain second filtrate.For the residue of second filtrate,, obtain the 3rd filtrate and the 4th filtrate according to repeating 2 times with above-mentioned same operation.Merge above-mentioned 4 kinds of filtrates, vacuum concentration to the proportion of concentrated solution is 1.05 (50 ℃), thereby obtains final extract.The weight of the described final extract that so obtains is 6.2kg.

Purifying process: above-mentioned final extract 6kg is dissolved in the water of this final half weight of extract under the condition of limit heating.Next, this extract solution is splined in the adsorption column that is filled with the D-101 type macroporous adsorbent resin of anticipating.This adsorption column of water eluting at first obtains the water elution liquid of 2 times of amounts of described fresh stem, and this adsorption column of the ethanol elution of reuse 20% obtains 20% ethanol, first eluent of 2 times of amounts of described fresh stem.Above-mentioned water elution liquid is repeated to adsorb once more-the such operation of eluting, obtain 20% ethanol, second eluent.Merge this 2 kind of 20% ethanol elution,, obtain containing the prepared product (embodiment 1) of phenethyl alcohol glycoside compounds by concentrated, dry.The weight of the prepared product that so obtains is 0.865kg.

Measure the content of echinacoside and acteoside by high-efficient liquid phase technique (HPLC).At this moment, the condition of HPLC is as follows: immobile phase is a C18 alkyl silane bonded silica gel, and mobile phase is methanol-0.15% aqueous acetic acid (30: 70), and flow velocity is 1 ml/min, and the detection wavelength is 330nm.

Take by weighing in 60 ℃ of vacuum echinacoside and acteoside after dry 24 hours, be dissolved in 50% the methanol, be mixed with reference substance solution (solute that contains 0.1mg in the 1ml solution).

The prepared product 50mg that limit ultrasonic Treatment, limit will contain the phenethyl alcohol glycoside compounds is dissolved in the 50% an amount of methanol in the 25ml volumetric flask.In above-mentioned solution, add 50% methanol again, be adjusted to the scale of 25ml.Precision is measured the about 1ml of this solution, puts in the volumetric flask of 10ml, adds 50% methanol until scale.Behind this solution of membrane filtration of 0.45 μ m, make sample solution.

Measure reference substance solution and the sample solution of 5 μ l respectively, inject high performance liquid chromatograph, measure the peak area of echinacoside and acteoside respectively.According to their its content of calculated by peak area.Its result is that the content of echinacoside in prepared product (embodiment 1) is that 37.5 quality %, the content of acteoside in prepared product (embodiment 1) are 9.0 quality %.

Use the pure product of echinacoside (Xinghui Medicine Industry Co. Ltd.'s manufacturing) among the embodiment 2, use the pure product of acteoside (Xinghui Medicine Industry Co. Ltd.'s manufacturing) among the embodiment 3.

Test example: the evaluation of hyaluronidase inhibitory activity

Test example 1: the evaluation of Cistanche Tubulosa extract (embodiment 1)

Method: add the 0.1ml enzyme activator in the 0.05ml enzymatic solution, placement is after 20 minutes down at 37 ℃, and adding the foregoing description prepared product 0.1ml after placing 20 minutes under 37 ℃, adds hyaluronic acid solution 0.25ml again, reacts 40 minutes under 37 ℃.Behind the NaOH 0.1ml of adding 0.4N reaction is stopped, adding boric acid solution 0.1ml then, heated 3 minutes down at 90 ℃.After the frozen water cooling, add right-dimethylaminobenzaldehyde of 3ml, after placing 20 minutes under 37 ℃, measure absorbance (585nm).The result as shown in Figure 1.

Test example 2 and test example 3: same, the echinacoside (test example 2) of embodiment 2 and the acteoside (test example 3) of embodiment 3 are estimated.Their result respectively as shown in Figures 2 and 3.

The effect of result and embodiment: as shown in Figure 1, for Cistanche Tubulosa extract, its hyaluronic acid enzyme inhibition shows the significant concentration dependency.In addition, as shown in Figure 2, the hyaluronic acid enzyme inhibition of the echinacoside of embodiment 2 also shows the significant concentration dependency in 1 μ g/ml-100 μ g/ml scope.Can determine thus: the hyaluronidase inhibiting part of the extract of embodiment 1 in organism (human body) is relevant with echinacoside.In addition, as shown in Figure 3, acteoside shows the hyaluronic acid enzyme inhibition in concentration is 3 μ g/ml-100 μ g/ml scopes.Can determine thus: the hyaluronidase inhibiting part of the extract of embodiment 1 in organism (mammal that comprises the people) is relevant with acteoside.By above result as can be seen, Cistanche Tubulosa extract and the echinacoside that is wherein contained thereof and acteoside have the hyaluronic acid enzyme inhibition in organism (mammal that comprises the people).

Enumerate the Formulation Example of Hyaluronidase inhibitor of the present invention below, but these Formulation Examples and be not used in qualification the present invention.

Formulation Example 1: chewing gum

Saccharum Sinensis Roxb. 53.0 weight %

Gum base 20.0

Glucose 10.0

Water maltosemalt sugar 16.0

Spice 0.5

Hyaluronidase inhibitor 0.5

100.0 weight %

Formulation Example 2: chewable tablet

Reductive water maltosemalt sugar 40.0 weight %

Saccharum Sinensis Roxb. 20.0

Glucose 20.0

Gelatin 4.7

Water 9.68

Sucus Vitis viniferae 4.0

Fructus Vitis viniferae local flavor 0.6

Pigment 0.02

Hyaluronidase inhibitor 1.0

100.0 weight %

Formulation Example 3: confection

Saccharum Sinensis Roxb. 50.0 weight %

Water maltosemalt sugar 33.0

Water 14.4

Organic acid 2.0

Spice 0.2

Hyaluronidase inhibitor 0.4

100.0 weight %

Formulation Example 4: Yoghurt (rigid soft)

Milk 41.5 weight %

Skimmed milk breast 5.8

Saccharum Sinensis Roxb. 8.0

Agar 0.15

Gelatin 0.1

Lactobacillus 0.005

Hyaluronidase inhibitor 0.4

The spice trace

Water is supplied

100.0 weight %

Formulation Example 5: refreshment drink

The liquid glucose 30.0 weight % of fructose glucose

Emulsifying agent 0.5

Hyaluronidase inhibitor 0.05

Spice is an amount of

Purified water is supplied

100.0 weight %

Formulation Example 6: cookie

Saccharum Sinensis Roxb. 76.4 weight %

Glucose 19.0

Sucrose fatty acid ester 0.050.2

Hyaluronidase inhibitor 0.5

Purified water 3.9

100.0 weight %

Formulation Example 7: soft capsule

Oleum Vitis viniferae 87.0 weight %

Emulsifying agent 12.0

Hyaluronidase inhibitor 1.0

100.0 weight %

Formulation Example 8: tablet

Lactose 54.0 weight %

Crystalline cellulose 30.0

Amylolysis thing 10.0

Fatty acid glyceride 5.0

Hyaluronidase inhibitor 1.0

100.0 weight %

Formulation Example 9: granule Oral preparation (pharmaceuticals)

Hyaluronidase inhibitor 1.0 weight %

Lactose 30.0

Corn starch 60.0

Crystalline cellulose 8.0

Polyvinylpyrrolidone 1.0

100.0 weight %

Formulation Example 10: pomade

Squalane 20.0 weight %

Cera Flava 5.0

Refining Jojoba oil 5.0

Glycerol 5.0

Glyceryl monostearate 2.0

Polyoxyethylene (20) sorbitan monostearate 2.0

Hyaluronidase inhibitor 2.0

Antiseptic is an amount of

Spice is an amount of

Purified water is supplied

100.0 weight %

Formulation Example 11: astringent

Ethanol 5.0 weight %

Glycerol 2.0

1,3 butylene glycol 2.0

Polyethylene oil ether 0.5

Sodium citrate 0.1

Citric acid 0.1

Hyaluronidase inhibitor 0.1

Purified water is supplied

100.0 weight %

Formulation Example 12: profit body breast

Macadimia nut oil 2.0 weight %

Myristic acid octyl group dodecyl ester 10.0

Methyl phenyl silicone 5.0

Tadenan 3.0

Stearic acid 3.0

Batilol 1.0

Glyceryl monostearate 1.0

Four oleic acid polyoxyethylene sorbitol 2.0

Hydrogen adds soybean phospholipid 1.0

Ceramide 0.1

Retinyl palmitate 0.1

Antiseptic is an amount of

Herba Centellae extract 1.0

Hyaluronidase inhibitor 1.0

1,3 butylene glycol 5.0

Purified water is supplied

100.0 weight %

Formulation Example 13: emulsion

Squalane 4.0 weight %

Vaseline 2.5

Spermol 2.0

Glycerol 2.0

Lipophilic glyceryl monostearate 1.0

Stearic acid 1.0

L-arginine 1.0

Hyaluronidase inhibitor 0.5

Potassium hydroxide 0.1

The spice trace

Purified water is supplied

100.0 weight %

Formulation Example 14: wash baths (aqueous)

Propylene glycol 50.0 weight %

Ethanol 20.0

Sodium sulfate 5.0

Hyaluronidase inhibitor 0.5

Lanoline 0.5

Shea oil 0.5

Pigment 1.5

Spice 22.0

100.0 weight %

Formulation Example 15: cat food

Semen Maydis powder 34.0 weight %

Wheat flour 35.0

Meat powder 15.0

Adeps Bovis seu Bubali 8.9

Sal 1.0

Qian fish extract 4.0

Hyaluronidase inhibitor 1.0

Taurine 0.1

Vitamins 0.5

Minerals 0.5

100.0 weight %

Formulation Example 16: dog food

Semen Maydis powder 30.0 weight %

Meat (Carnis Gallus domesticus) 15.0

Defatted soybean 10.0

Wheat flour 25.0

Bran bran class 5.0

Hyaluronidase inhibitor 5.0

Animal raw fat 8.9

Oligosaccharide 0.1

Vitamin 0.5

Mineral 0.5

100.0 weight %

As mentioned above, the present invention can suppress the activity of hyaluronidase, helps to make biological intravital hyaluronic acid to keep stable, can prevent the aging of skin thus effectively, but also can be as antiinflammatory and anti-allergic agent etc.

Concentration in the accompanying drawing (μ g/mL); Hyaluronic acid enzyme inhibition rate (%)

Claims (10)

1, a kind of Hyaluronidase inhibitor, it is characterized in that: active ingredient is the phenethyl alcohol glycoside compounds, described phenethyl alcohol glycoside compounds comprises at least a in echinacoside and the acteoside, or described phenethyl alcohol glycoside compounds comprises echinacoside and acteoside.

2, a kind of Hyaluronidase inhibitor is characterized in that: effective ingredient is the extract of Orobanchaceae plant, contains the phenethyl alcohol glycoside compounds in the described extract as effective ingredient.

3, according to the Hyaluronidase inhibitor described in the claim 2, it is characterized in that, contain at least a in echinacoside and the acteoside in the described extract, or contain echinacoside and acteoside in the described extract as described phenethyl alcohol glycoside compounds as described phenethyl alcohol glycoside compounds.

4, a kind of resisting age of skin agent is characterized in that, contains any described Hyaluronidase inhibitor in the claim 1 to 3 in this resisting age of skin agent.

5, a kind of skin preparations for extenal use that can be used for comprising people's mammal, this skin preparations for extenal use with the resisting age of skin agent described in the claim 4 as effective ingredient.

6, a kind of is the drinking food product of effective ingredient with the resisting age of skin agent described in the claim 4.

7, a kind of is that the mammal of effective ingredient is used feedstuff with the resisting age of skin agent described in the claim 4.

8, a kind of antiinflammatory is characterized in that: contain any described Hyaluronidase inhibitor in the claim 1 to 3 in this antiinflammatory.

9, a kind of medicine that can be used for comprising people's mammal, wherein this medicine is an effective ingredient with the antiinflammatory described in the claim 8.

10, a kind of skin preparations for extenal use that can be used for comprising people's mammal, wherein this skin preparations for extenal use is an effective ingredient with the anti-antiinflammatory described in the claim 9, and antiinflammatory is the drinking food product of effective ingredient or uses feedstuff for the mammal of effective ingredient.

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