CN1372919A - Use of bean products for treating exogenous invasion - Google Patents

Abstract

The present invention features legume products having trypsin inhibitory activity and reduced microbial content, methods of decontaminating such legume products, compositions containing such legume products, and the topical application of such legume products or compositions to skin, nails, and hair.

Description

Use bean products for treating exogenous invasion

Field that the present invention belongs to

The present invention relates to bean products, comprise the topical compositions of described bean products, and the preparation of these products and application.

Background of invention

The fruit of beans contains high-caliber protein, lipid and sugar usually.Therefore, bean fruits such as Semen sojae atricolor, and the compositions that comprises bean fruits, all being considered to has the very product of high nutritive value to the people.Bean fruits also contains the material of Profilin enzymatic activity usually.For example, two kinds of albumen enzyme inhibitors were promptly separated from Semen sojae atricolor as far back as the forties in 20th century, and (soybean trypsin inhibitor is STI) with Bowman-Birk trypsin inhibitor (BBI) for Kunitz type trypsin inhibitor.List of references is seen Birk, Int.J.Pept.Protein Res.25:113-131 (1985) and Kennedy, Am.J.Clin.Neutr.68:1406S-1412S (1998).

STI suppresses tryptic proteolytic activity by forming stable stoichiometric complex.Referring to Liu, K., the nutritive value of chemistry and soybean component.At " Semen sojae atricolor, chemistry, technology and application " 32-35 page or leaf (Aspen publishers, Inc., Gaithersburg, MD, 1999).STI contains 181 amino acid residues, by two disulfide bond bridgings, and is roughly sphere.Referring to Song etc., J.Mol.Biol.275:347-63 (1998).

BBI is the protein of a 8k-Da, suppresses trypsin and Chymotrypsin in different reaction site.Referring to the Billings pen, Pro.Natl.Acad.Sci.89:3120-3124 (1992).STI and BBI only are found in soybean seed, and do not find at other position of bean plant.Referring to Birk, Int.J.Pept.ProteinRes.25:113-131 (1985).

But, because its natural origin, have a large amount of microorganisms in the appearance of bean fruits such as Semen sojae atricolor.Thereby purification process is as heating, organic solvent extraction, and the concentration that the purification of high shear force can reduce microorganism makes it when the people uses, when using, safer such as skin protection.Yet have been found that these processing procedures can make active substance degeneration in the Semen sojae atricolor usually, cause those application on human skin, hair and fingernail to be had the destruction (as the reduction of protease inhibiting activity) of the important biological effectiveness of beauty treatment and therapeutical effect.And these processing procedures also can cause the instability of soybean prod, produce offensive odour and change in color.Thereby just there is exploitation in the biological effectiveness of not destroying these soybean prods, to reduce the business demand of the micro organism quantity of soybean prod.

The purpose of this invention is to provide a kind of soybean prod (for example, can be used as the active component of the care composition of skin, hair or fingernail), this product has reduced micro organism quantity and has kept its protease inhibiting activity.Another object of the present invention provides skin, hair and the fingernail care composition that comprises described soybean prod and other optional active component.

The present invention relates to include the bean products that has reduced micro organism quantity and kept its effective biologic activity, the acquisition methods of this product and the application in cosmetic composition.

Summary of the invention

The present invention relates to have the bean products of the micro organism quantity of trypsin inhibition activity and reduction, purify the method for above-mentioned soybean prod, and the compositions that comprises above-mentioned soybean prod.In a preferred embodiment, this bean products is a soybean prod.

The compositions that the invention still further relates to bean products or contain described bean products is in the health of safeguarding skin, hair and fingernail, and local use the in the disease of prevention or treatment skin, hair and fingernail, described prevention or treatment comprise but are not defined as: the steadiness of regulating skin, hair and fingernail; Cleaning skin, hair and fingernail; Reduce or delay the growth of hair and fingernail; Stretching or blast hair; Treatment or prevention acne; Regulate the color pool of skin, hair and fingernail; Regulate the stricture of vagina reason of skin; Regulate the wrinkle of skin; Handle the external infringement of skin, hair and fingernail; And beautify skin, hair and fingernail.

Other features and advantages of the present invention will be below description details and claims in introduce.

Detailed Description Of The Invention

Should be sure of that those of ordinary skills can maximally utilise the present invention on the basis of present specification.Following embodiment preferred only is used for description and interpretation, and limits the present invention never in any form.

Except as otherwise noted, technology of the present invention and scientific terminology all are according to the common meaning of understanding of those of ordinary skills.And all publications of mentioning here, patent application, patent and other data are all used for referencial use.In this article, unless otherwise indicated, all percentage ratios all are percentage by weights.

Here " trypsin inhibition activity " mentioned refers to the ability of the bean products inhibition tryptic activity of 0.1% (w/w) concentration, measures by following examples 2 described measuring methods.The tryptic activity that bean products of the present invention in one embodiment has at least about 15% suppresses ability.Further in the preferred embodiment, bean products has an appointment 25% at least, for example suppresses ability at least about 50% tryptic activity.

Here " the mercaptan retentive activity " mentioned refers to the inhibition ability of the bean products of 0.1% (w/w) concentration to the inductive mercaptan loss of cigarette, measures by following example 3 described measuring methods.In one embodiment, bean products of the present invention has the mercaptan retentive activity at least about 75%.In the further preferred embodiment, bean products has at least about 90%, for example at least about 95% mercaptan retentive activity.

Here " content of microorganisms " mentioned refers to antibacterial, fungus and the saccharomycetic quantity that is present in bean products.The example of measuring the method for content of microorganisms comprises, but be not defined as, at " Official Methodsof Analysis of AOAC International ", Patricia Cunniff chief editor, the 16 edition, the 5th revision, AOAC 986.23 methods of pointing out among 1999 (the AOAC International); Or at " Official Compendiaof Standards, USP 24 USP/NF 19 ", American Pharmacopeia agreement, Inc., the method that 2000 (Board of Trustees, American Pharmacopeia agreements) are pointed out.

" unallowed content of microorganisms " meaning is meant antibacterial, fungus and the saccharomycetic quantity in the bean products of being present in that is pernicious to people, comprise, but be not limited to, coliform group, escherichia coli, Salmonella, thermophilic spore, bacillus, enterococcus, staphylococcus, streptococcus faecalis and those are " Disinfection, sterilization; and preservation " the 4th edition, Seymour S.Block, and pp.887-888 (1991, Lea ﹠amp; Febiger, Malvern, PA) microorganism of being mentioned in the literary composition.

Here " topical application " mentioned refers to direct coating or sprawls in skin, available hands or such as brush expand cotton, walk applicators such as pearl or aerosol apparatus and apply.

Here " it is acceptable to make up " mentioned is meant that those are suitable for contacting with tissue (as skin), do not have the product or the material of unsuitable toxicity, incompatibility, unstability, zest, anaphylaxis etc.

Here " topical carrier " mentioned refers to one or more solids that are suitable for the mammal local application or liquid filling diluent.The example of topical carrier includes, but are not limited to water, wax, oil, softening agent, emollient, thickening agent, gel, and their mixture.

Here " the steadiness adjustment " mentioned refers to and strengthens steadiness or elasticity, prevents or treat the lax, loose or sagging of skin, hair and fingernail.The steadiness of skin or elasticity are the mensuration that should be used for by the epidermis analyzer.Referring to Non-invasive Methods and the Skin handbook, eds.J.Serup ﹠amp; G.Jemec, Chapter 14.3 (1995).Skin steadiness or elastic forfeiture can be that multiple factor causes, and include but not limited to wear out, the using of exogenous injury or skin, hair and nail cosmetic.

Here " adjustment of color and luster " mentioned refers to the outward appearance (as the painted infringement of blast, deepen the palor color, and/or the liquid skin colour being even) of blast and/or deepening skin, hair and fingernail.

Here " delaying or weaken the growth of fingernail " of mentioning refers to the growth rate that delays or weaken fingernail.

Here " delaying or weaken the growth of hair " of mentioning refers to and delays or weaken the growth rate of hair and/or dry width, includes but not limited to reduce the visibility and the outward appearance (as arm, shank and facial hair) of hair.

Here " cleaning " mentioned refers to from skin, hair and nail surface and removes foul and/or greasy dirt.

Here " the adjusting texture " mentioned refers to lump and the breach of eliminating skin, hair and nail surface, makes smooth surface, includes but not limited to make skin appearance smoothly even.

Here " regulating the wrinkle of skin " of mentioning refers to the forming process that prevents, delays or reverse the trickle wrinkle of skin, includes but not limited to reduce the visibility of chill mark.

Here " the treatment exogenous injury " mentioned refers to and prevents or alleviate because the damage that the exogenous injury of skin, hair and fingernail that physics contact, chemistry contact or temperature are brought causes.The example of exogenous injury includes but not limited to from following damage to skin, hair and fingernail: the use of chemicals such as abluent (as containing the skin of surfactant and the abluent of hair) and cosmetics; The damage that physics contact causes is as having a shave and cutting etc.; Environmental factors is brought damage as ozone, waste gas, pollution, chlorine and is contained chlorine compound and the infringement of chemistry such as smoking, blows to wait mechanically to encroach on the infringement that cold and hot temperature is brought.Exogenous injury includes but not limited to oxidation and/or nitrosation destruction and the modification to lipid, saccharide, peptide class, protein, nucleic acid and vitamin to the influence of skin, hair and fingernail.The influence of exogenous injury also includes but not limited to the forfeiture of cell survival, the forfeiture of cell function and the change of change, gene and/or protein expression.

Here " the safe and effective amount " mentioned refers to used chemical compound or compositions (as bean products) enough produces positive modification to the situation that is conditioned or handles, but can not cause the consumption of serious adverse.The safe and effective amount of chemical compound or compositions can be along with quilt treatment situation final user age and health, the seriousness of preventing/treating situation, the persistent period of treatment, the situation of other treatment, the characteristic of used material or product/compositions, the characteristic of used cosmetics acceptable carrier and the difference of some other factors and change.

Bean products

" bean products " refers to from the deutero-material of bean fruits.Bean derives from leguminous plant usually, and rimose fruit is all arranged, as Semen Viciae fabae, Semen Pisi sativi or Radix Crotalariae szemoensis.The example of bean includes but not limited to Semen sojae atricolor, Radix Crotalariae szemoensis, Semen Pisi sativi and Semen arachidis hypogaeae.

Bean products can comprise whole bean fruits (as bean fruits is crushed into powder), or only uses the part (as the extract of bean fruits) of beans.Bean products can be the form of fluid (as the mixture of bean fruits and water) or solid (as the powder of bean fruits).When being fluid form, term " bean products " refers to the solid constituent in the fluid that derives from bean.

Compositions of the present invention comprises the bean products (as soybean prod) of safe and effective amount.In a specific embodiment, it is about 0.001% to 50% that compositions comprises, preferred about bean products of 1% to 30% (as soybean prod).

Soybean prod

" soybean prod " refers to the material derived from Semen sojae atricolor.Soybean prod can only comprise the part (Semen sojae atricolor powder that reduces as soybean extract such as lipid, or filterable bean milk) of Semen sojae atricolor, also can comprise whole Semen sojae atricolor (as the Semen Glycines powder end of pulverizing).Soybean prod can be fluid (as a bean milk), or solid (as powdered soybean or Semen sojae atricolor milk powder).When being fluid form, term " soybean prod " refers to the solid constituent in the fluid that derives from Semen sojae atricolor.

In a specific embodiment, soybean prod is a powdered soybean.Powdered soybean can make by rolling exsiccant Semen sojae atricolor.In a specific embodiment, powdered soybean has and is lower than 10 microns mean diameter, as less than 1 micron.In a specific embodiment, the moisture of powdered soybean is less than 10%, as less than 5%.In a specific embodiment, powdered soybean is cryodesiccated.

In a specific embodiment, soybean prod is bean milk or bean milk powder.Bean milk is the solid that derives from Semen sojae atricolor and the mixture of water, all or part of insoluble matter of filtering in the mixing.The bean milk powder is to obtain behind the bean milk evaporate to dryness, in a specific embodiment, is lyophilization or spray-dired form.The process of preparation bean milk includes but not limited to following three processes.At first, Semen sojae atricolor is put into water and absorb moisture; Expansible bean or pea are rolled subsequently, and add remaining water; Filtering mixt is removed insoluble residue.The second, bean milk can prepare from powdered soybean.Powdered soybean thoroughly mixes (as mixing at least one hour) with water, then remove by filter insoluble residue.The 3rd, can also be by adding entry and the bean milk powder is reassembled into bean milk.In a specific embodiment, bean milk accounts for 1% to 50% (as 5% to 20%) of soy bean solid weight.

The antimicrobial treatment of bean products

As mentioned above, the microorganism of higher concentration is contained on the bean fruits surface usually.Thereby when previous people used, bean products all will reduce or eliminate these microorganisms through handling.The microbial count of bean products of the present invention is less than 10000 bacterium colony units of every gram (cfu) in a specific embodiment.In another embodiment, the microbial count of soybean prod of the present invention is approximately less than the every gram of 1000cfu (for example less than the every gram of 100cfu).

In a specific embodiment, in the bean products of the present invention unallowed microbial count less than the every gram of 300cfu, for example, less than the every gram of 150cfu.In further embodiment, bean products of the present invention unallowed microbial count at least 1 gram (for example at least 10 grams) product reaches the level that survey does not go out.

In a specific embodiment, bean products is exposed in the radiation of gamma ray.In further embodiment, bean products is to be exposed under the radiation that intensity is 2 to 30KGy gamma ray, and for example the radiant intensity of gamma ray is 5 to 10KGy.The applicant is unexpected to find such processing in the content of microorganisms that has reduced bean products, has kept its biological activity (suppressing active as serine protease).The applicant also finds to handle the advantage that bean products can keep the cosmetics of this product with gamma ray, as the maintenance natural colored, and does not bring tangible stink.

Other same microbial treatment method of the protease inhibiting activity of bean products that keeps can be used separately, or can unite use with gamma ray radiation, these processing methods include but not limited to the X-ray, high-power electron beam or proton beam, ultraviolet radiation, hydrostatic pressure and adding have the chemical reagent of antimicrobial acivity, and their combination.The complete documentation of a method that reduces micro organism quantity is " sterilization, sterilization are preserved " book the 4th edition, Seymour S.Block, pp.887-888 (1991, Lea ﹠amp; Febiger, Malvem illustrates in PA).

The applicant finds to use heat-treating methods can cause the forfeiture of protease inhibiting activity, and very careful when handling.For example, the applicant find 100 ℃ down the heating bean milk as long as 10 minutes can (at 4 ℃ time) be reduced to 46% from 86% with the protease inhibiting activity of bean milk.The applicant also finds to heat the variation that bean milk can cause product color and taste.

Topical compositions

The used topical composition of the present invention comprises that those are suitable for the combination that local skin is used.In a specific embodiment, compositions comprises soybean prod and the acceptable topical carrier of cosmetics.In a specific embodiment, the acceptable topical carrier of cosmetics accounts for 50% to 99.99% (for example from 80% to 95%) of composition weight.

Compositions can be made various product type, include but not limited to lotion, white Emulsion, gel, club, spray, shaving cream, ointment, cleaning washing liquid and solids wash rod, shampoo, unguentum, powder, mousse, shaving cream, liniment, patch, nial polish, trauma dressing and cohesive bandage, hydrogel, thin film and such as the cosmetics of foundation cream, mascara and lip pomade etc.These product forms can contain the acceptable topical carrier of several cosmetics, include but not limited to solution, emulsion (as microemulsion and millimicro emulsion), gel, solid and liposome.It below is the infinite example of these carriers.Other carrier can be obtained according to a conventional method by those of ordinary skills.

Topical composition of the present invention can be made solution.Solution generally includes aqueous solvent (as 50%-99.99%, or the acceptable aqueous solvent of the cosmetics of 90%-99%).

The topical composition that the present invention uses can be made the solution that contains emollient.Such compositions preferably comprises the emollient of about 2%-50%.Here, " emollient " is meant that those can be used to prevent or alleviate drying, and reaches the material of protection skin effects.Many suitable emollient all are known, and can use at this.Sagarin, Cosmetics, Science and Technology, the 2nd edition, Vol.1, pp.32-43 (1972), with international cosmetic active ingredient dictionary and handbook, eds.Wenninger and McEwen, pp.1656-61,1626, and 1654-55 (The Cosmetic, Toiletry, and Fragrance Assoc., Washington, D.C., the 7th edition, 1997) put down in writing the example of many suitable above-mentioned substances in (hereinafter to be referred as " ICI Handbook ").

Lotion can be from above-mentioned formulations prepared from solutions.Lotion contains a kind of emollient of about 1%-20% (as, about 5%-10%) and the about water of 50%-90% (as, about 60%-80%) usually.

Product another kind of the present invention can be Emulsion from the dosage form of formulations prepared from solutions.Emulsion contains the emollient of the 5%-50% that has an appointment (as, about 10%-20%) and the about water of 45%-85% (50%-75% according to appointment) usually.

Also having a kind of can be ointment from the dosage form of formulations prepared from solutions.Ointment contains a kind of simple substrate such as animal oil, vegetable oil or semisolid hydro carbons usually.Ointment can contain the emollient of about 2%-10%, adds the intensifier of about 0.1%-2%.Intensifier or thickening agent commonly used can be at Sagarin, Cosmetics, and Scienceand Technology, the 2nd edition, Vol.1 finds among pp.72-73 (1972) and the ICI Handbook pp.1693-1697.

Topical composition of the present invention can be made emulsion.If carrier is a kind of emulsion, should contain the 1%-10% that the has an appointment emulsifying agent of (as, about 2%-5%).Emulsifying agent can be nonionic, anion or cationic.The emulsifying agent that is fit to is in U.S. Patent No. 3755560,4421769, McCutcheon ' s Detergents andEmulsifiers, North American Edition, pp.317-324 (1986) and ICI Handbook, open among the pp.1673-1686.

Lotion and cream can be made emulsion.Usually such lotion contains the emulsifying agent of about 0.5%-5%.Such Emulsion contains the emollient of about 1%-20% (5%-10% according to appointment) usually; The water of about 20%-80% (30%-70% according to appointment); Emulsifying agent with about 1%-10% (2%-5% according to appointment).

The single-phase emulsion skin care formulation of oil-in-water type and water-in-oil type as lotion and Emulsion, all is that cosmetic field is known, and uses in the present invention.Multiphase emulsion composition, as the W/O/W type, open and also use in the present invention in U.S. Patent No. 4254105 and 4960764.Usually, above-mentioned single-phase and heterogeneous emulsifying agent all contains water, emollient and emulsifying agent as necessary component.Topical composition of the present invention can also be made gel (as making hydrogel with suitable gel).Include but not limited to natural gum, acrylic acid and acrylate polymer and copolymer, cellulose derivative (as hydroxy methocel, hydroxyethyl-cellulose) as the suitable gel of aqueous gel.The gel of the oleogel (as mineral oil) that is fit to includes but not limited to hydrogenation butylene/ethylene/styrene copolymer, and hydrogenation of ethylene/propylene/styrene copolymer.Above-mentioned gel contains the above-mentioned gel of percentage by weight for about 0.1%-5% usually.

Topical compositions of the present invention can also be solid dosage forms (as the bar of wax system, soap composition, powder, or brush).

Liposomal formulation also is a compositions of the present invention.Liposome can be a monolayer, and the liposome of multilamellar and few layer can comprise or not comprise phospholipid.Preparing such compositions at first is with Tangeretin and phospholipid, and as dipalmitoyl phosphatidyl choline, cholesterol and water are according to Mezei ﹠amp; Gulasekharam, " Liposomes-ASelective Drug Delivery System for the Topical Route of Administration; Gel DosageForm ", Journal of Pharmaceutics and Pharmacology, Vol.34 (1982), the described method combination of pp.473-474, or modify.The epidermis lipid that forms the suitable compositions of liposome can substitute phospholipid.Be that liposome is blended in (as gel or O/w emulsion) on a kind of above-mentioned carrier then, to obtain the liposome dosage form.Other compositions and local application liposome medicinal at Mezei, M., " Liposomesas a Skin Drug Delivery System ", Topics in Pharmaceutical Sciences (D.D.Breimerand P.Speiser, eds.,) Elsevier Science Publishers B.V., New York, N.Y., 1985, pp.345-358 has detailed description in PCT Patent patent application WO9631194 and the U.S. Patent No. 5260065.

Topical composition of the present invention can contain multiple additional oil soluble material and/or water-soluble substances except that aforementioned component, they can be used for skin, hair and fingernail according to a conventional method with the level that prior art is established and be used for compositions.

Additional cosmetic activating agent

In a specific embodiment, topical composition can further contain another kind of cosmetic activating agent beyond bean products.Here " cosmetic activating agent " refers to skin, hair and fingernail have the material of beauty treatment or therapeutical effect, as blast reagent, deepening reagent (as from tanned reagent), anti-acne reagent, delustering agent, antimicrobial, antiinflammatory, antifungal, antiparasitic, the external application analgesic agent, sunscreen, light protective agent, antioxidant, keratolytic, detergent/surfactant, humidizer, nutrient, vitamin, energy booster, anti-diaphoretic, astringent, deodorizer, the hair remover, firming agent, anti-hardening agent and skin, hair and fingernail conditioner.

In a specific embodiment, these reagent can be selected from and include but not limited to hydroxy acid, benzoyl hydroperoxide, thioresorcin, ascorbic acid, D-panthenol, hydroquinone, methoxy cinnamic acid octyl group ester, titanium dioxide, salicylic acid octyl group ester, heliophan, avobenzone, poly-bakelite, carotenoid, free radical scavenger, from executing agent for capturing, biostearin such as retinol and retinol cetylate, ceramide, polyunsaturated fatty acid, essential fatty acid, enzyme, enzyme inhibitor, mineral, hormone such as estrogen, steroid such as hydrocortisone, the 2-dimethylaminoethanol, mantoquita such as copper chloride, cupric peptide such as Cu:Gly-His-Lys, coenzyme Q10, as those disclosed peptides in PCT patent application WO0015188, thioctic acid, aminoacid such as proline and tyrosine, vitamin, lactobionic acid, S-acetyl-coenzyme-A, nicotinic acid, riboflavin, vitamin B1, ribose, electron transfer agent such as NADH and FADH2, and other plant extract such as Aloe, their derivant and composition thereof.The content of the active cosmetic agent in the present composition accounts for about 0.001%-20% of composition weight usually, and for example about 0.01%-10% further is 0.1%-5% according to appointment.

The example of vitamin includes but not limited to vitamin A, and vitamin B is serial as vitamin B3, vitamin B5, vitamin B12, vitamin C, vitamin K, and vitamin E and derivant thereof.

The example of hydroxy acid includes but not limited to glycolic, lactic acid, and malic acid, salicylic acid, citric acid, and tartaric acid etc., referring to european patent application No.273202.

The example of antioxidant includes but not limited to water solublity antioxidant such as mercaptan compound and derivant, (as sodium metabisulfite and N-acetylcystein), thioctic acid and dihydrolipoic acid, resveratrol, lactoferrin, and ascorbic acid and ascorbic acid derivates (as sodium L-ascorbate-2-phosphate and vitamin C polypeptide).The oil-soluble antioxidant that is applicable to compositions of the present invention comprises but is not limited to Yoshinox BHT, biostearin (as retinol and retinol cetylate), vitamin E (as tocopherol acetate), tocotrienol, ubiquinone.The natural extract that contains antioxidant that is suitable for the present composition includes but not limited to flavone, and the extract of isoflavone and derivant (as trihydroxy-isoflavone and diadzein) contains resveratrol extract and other.Natural extract comprises Semen Vitis viniferae, green tea, Cortex Pini, and propolis.The example of other antioxidant is referring to ICIHandbook 1612-13 page or leaf.

Other material

Other different material also can be used for compositions of the present invention.These comprise wetting agent, protein and polypeptide, antiseptic, and alkaline reagent.These reagent are at ICI Handbook, and are open among the pp.1650-1667.Compositions of the present invention also contains chelating agen (as EDTA) and antiseptic (as parabens).The suitable antiseptic and the example of chelating agen are at ICI Handbook, and be open among pp.1626 and the 1654-55.In addition, the topical composition here contains the cosmetic adjuvant usually, as dyestuff, and opacifier (as titanium dioxide), pigment and flavouring agent.

Mineral water

Bean products of the present invention (as bean milk) and compositions can be used mineral water in preparation process.In a specific embodiment, the mineralization degree of mineral water is about 200mg/L (300mg/L is to about 1000mg/L according to appointment) at least.In a specific embodiment, mineralized water comprises at least about the calcium of 10mg/L and/or at least about the magnesium of 5mg/L.

The compositions and the preparation that contain compositions of the present invention all can be prepared by known routine techniques by those of ordinary skills.

Embodiment 1: the gamma ray radiation of bean products is handled

The applicant finds that disinfecting action (as gamma ray radiation) is preceding to contain high-caliber microorganism to soy milk powder carrying out, and mostly is every gram 50000cfu most.Find that also but there is the unallowed microorganism of detection level in such product, as streptococcus faecalis, concentration is up to every gram 20000cfu.

The applicant places the soy milk powder (as restraining about 200 kilograms from about 1) of varying number under the gamma ray from 1KGy to 16KGy.The needed dosage that makes total content of microorganisms be lower than the every gram of 100cfu approximately is 10KGy.Making streptococcus faecalis reduce that 1 logarithmic dosimetry goes out is about 3KGy, makes 10 to restrain content of microorganisms in the soy milk powder samples and remain on the required radiation dose of level that survey do not go out and be about 5KGy.But, how much be used to shine the amount of radiation of the gamma ray of bean products finally will deciding by content of microorganisms and the soybean prod of handling like this.

The trypsin inhibition activity of embodiment 2 bean products

Fluorescence casein peptide segment is induced cracked inhibitory action reagent to contain at trypsin and is used EnzChex ^TMThe protease assay test kit is measured (EnzChex according to the method that manufacturer instructs ^TMProtease assay test kit product information, 3/15/99 revision; Molecular Probes, Eugene OR).Briefly, the at first dilution in 1X digestion buffer (in test kit, preparing) of different bean products, and under different concentration with the 1000 trypsin Sigma of unit that are dissolved in the 1X digestion buffer, St.Louis, MO) cultivation together.Pure serpin (soybean trypsin inhibitor, from Sigma, St.Louis, MO) 0.1%, 0.01% and 0.001%w/v concentration as positive control.Then, 1.0mg/mlBODIPY FL casein storing solution obtains by add the 0.2ml deionized water to the bottle that substrate (preparing) is provided in test kit, and then, making ultimate density in the digestion buffer is 10 mcg/ml.Add or do not add under the situation of test substances, cultivate after one hour under trypsin and the BODIPY fluorescence casein substrate room temperature, fluoremetry (excitation wavelength 485nm, emission wavelength 530nm) is to pass through SpectraMax ^(Molecular Devices Corporation, Sunnyvale CA), use Softmax to Gemini microtitration plate reader ^Pro3.0 software (Molecular Devices Corporation) is finished.Each test is all triplicate, does twice at every turn.

This measuring method can be used for the measurement with the soybean prod of seven kinds of distinct methods processing.Example A be the Semen sojae atricolor that is crushed into powder (sunlight food company, Taipei, R.O.C).Example B is the powder with the example A of the gamma ray radiation processing of about 8-15KGy.Example C be the powdered soybean that is extracted out of oils and fats (from Central Soya Company Inc., Fort Weyne, the Soyafluff  of IN).Example D is the bean milk powder, and (Devansoy Farms, Carroll Iowa) and with the gamma ray radiation of 7-9KGy handle this powder by filtration, heating, spray drying by peeled soybeans and water.Example E is the bean milk powder, and this powder is by Semen sojae atricolor being mixed with water, mixture heated is spent the night, and adds to mix behind the 1,3 butylene glycol to obtain (from Japanese Ichimaru Pharcos CO., Ltd, the Flavosterone of Gifu) again.Example F is the bean milk powder, and this powder is by Semen sojae atricolor being mixed with water, mixture heated is spent the night, and obtains (from Japanese Ichimaru Pharcos CO., Ltd, the Flavosterone of Gifu) after adding ethanol again.Example G is soybean protein extract (from French Laboratories Serobiologiques S.A., the Vegeseryl HGP LS 8572 of Pulnoy).All these soybean prods all (Sigma) compare with soybean trypsin inhibitor (STI).

Different bean products pass through MicrosoftExcel to the inhibiting percent of trypsin cracking of substrate ^TMCalculate and be recorded in table 1.Table 1

Test products	Concentration	The % trypsin inhibition capacity
			????STI	????0.01	????43.0
????STI	????0.1	????76.1
			Example A	????0.01	????32.8
Example A	????0.1	????67.1
			Example B	????0.01	????31.5
Example B	????0.1	????67.2
			Example C	????0.01	????22.7
Example C	????0.1	????36.2
			Example D	????0.01	????8.92
Example D	????0.1	????17.4
			Example E	????0.01	????7.83
Example E	????0.1	????10.8
			Example F	????0.01	????4.87
Example F	????0.1	????5.99
			Example G	????0.1	????6.85

As shown in table 1, STI can suppress the inductive cracking of trypsin in the dose response mode.Example A, powdered soybean has significant trypsin inhibition activity equally.The gamma radiation of other powdered soybean (as example B) is made us unexpectedly its trypsin inhibition activity of not appreciable impact when reducing micro organism quantity.Heat and/or extract the product as example C-G of processing, but obviously reduced the trypsin inhibition activity of powdered soybean.

The mercaptan retentive activity of embodiment 3 bean products

Powdered soybean prevent by the ability of the inductive mercaptan of flue dust loss estimate at human body normal skin fibroblast (Clonetics, San Diego, CA).Mercaptan mainly is glutathion, is the part of endogenous cell anti-oxidative defense system.Glutathion is redox cushion, thereby can keep the balance of oxidant and antioxidant.Glutathion still is the preferred substrates of some enzymes, as glutathion peroxidase (decomposition of hydrogen peroxide) and glutathione s-transferase (a kind of important separate toxenzyme).Referring to A.Meister, Cancer Res.54:1969s-1975s (1994).

The skin antioxidant (enzymatic and non-enzymatic) that comprises glutathion promptly can be ineffective in the presence of ultraviolet or ozone.Referring to M.J.Connor and L.A.Wheeler, Photochem.Photobiol.46:239-246 (1987) and R.M.Tyrrell and M.Pidoux, Photochem.Photobiol.47:405-412 (1988).In cell culture model, the glutathion inside cell of low content (GSH) low-level causes the higher sensitivity to ultraviolet radiation.Can prevent the photic damage of ultraviolet in rat skin surface local administration of cysteine derivant to skin; This effect is relevant with the synthetic increase of glutathion.Referring to L.T.van den Broeke and G.M.J.Beijersbergen van Henegouwen, J.Photochem.Photobiol.B Biol.27:61-65 (1995); K.Hanada, etc., J.Invest.Dermatol.108:727-730 (1997); And D.P.T. Steenvoorden, etc., Photochem.Photobiol.67:651-656 (1998).Therefore, glutathion is main endogenous antioxidant, has the higher respond to environmental change, can regulate skin color and wrinkle, can be used for handling exogenous injury.

In this experiment, contact or simulation flue dust environment (1 medicated cigarette, BASIC Full Flavor100 ' s cigarette, Philip Morris, Richmond VA) or use before the placebo simulation process 10 minutes, is seeded in Transwell insert (the Corning Costar of 24-hole form, Cambridge, MA) the normal newborn skin fiber cell of the human body in was cultivated 24 hours in comprising the bean milk powder of variable concentrations (gamma ray radiation with about 5KGy is handled).Before flue dust environment contact, remove the culture medium on the insert of soybean prod, (Life Technologies, Gaithersbury MD) give a baby a bath on the third day after its birth time, and the culture medium under the insert is contacted with cigarette to use Dulbecco ' s phosphate buffered saline(PBS).After the processing, at once cell was cultivated 24 hours with previous culture medium again, cell reuse Dulbecco ' s phosphate buffered saline(PBS) is washed five times, by in cell, adding monobromobimane (the Molecular Probes of 60 μ M, Eugene, OR USA) and after cultivating 30 minutes under 37 ℃ reads fluorescence to measure intracellular mercaptan.If mercaptan exists, then monobromobimane shows fluorescence.Fluorescence measurement is by using CytoFluor ^(MA USA) measures under following optical filtering condition: excitation wavelength 360nm, emission wavelength 460nm the fluorescent screen reader for PerSeptive Biosystems, Framingham.

This experimental result sees Table 2.Table 2

	Bean milk powder concn (weight %)	Mercaptan (comprises smokeless group mercaptan percent; Mean ± S.E.M.)
			There is not cigarette	????0	????100±6.71
Cigarette (10 minutes) is arranged	????0 ????0.5 ????1 ????2	????65.38±7.16 ????91.24±14.25 ????95.39±4.52 ????106.92±17.06

The result shows that the bean milk powder of handling with gamma ray radiation makes us unexpectedly having the protective effect (data are expressed as the meansigma methods of numerical value ± three an independent repeated experiments) of the mercaptan loss that the antagonism cigarette causes

Be appreciated that detailed description of the present invention is to be used to illustrate the present invention, but do not limit scope of the present invention, scope of the present invention is determined by the scope of additional claims.Other feature, advantage, and modify all in the scope of claims.

Claims (32)

1, a kind of compositions that is used to handle skin, fingernail or hair exogenous injury, said composition contains soybean prod.

2, the compositions of claim 1, wherein said exogenous injury are that the chemistry contact causes.

3, the compositions of claim 1, wherein said exogenous injury are that the physics contact causes.

4, the compositions of claim 2, wherein said exogenous injury is to use cleaning agent to cause.

5, the compositions of claim 3, wherein said exogenous injury are the damages of shaving and causing.

6, the compositions of claim 1, wherein said compositions are the exogenous injurys that is used to handle skin.

7, the compositions of claim 2, wherein said compositions are the exogenous injurys that is used to handle skin.

8, the compositions of claim 3, wherein said compositions are the exogenous injurys that is used to handle skin.

9, the compositions of claim 4, wherein said compositions are the exogenous injurys that is used to handle skin.

10, the compositions of claim 5, wherein said compositions are the exogenous injurys that is used to handle skin.

11, the compositions of claim 1, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

12, the compositions of claim 2, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

13, the compositions of claim 3, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

14, the compositions of claim 4, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

15, the compositions of claim 5, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

16, a kind of method of handling skin, fingernail or hair exogenous injury, this method comprises the local application soybean prod.

17, the method for claim 16, wherein said exogenous injury are that the chemistry contact causes.

18, the method for claim 16, wherein said exogenous injury are that the physics contact causes.

19, the method for claim 17, the injury that wherein said exogenous injury is to use cleaning agent to cause.

20, the method for claim 18, wherein said exogenous injury are the damages of shaving and causing.

21, the method for claim 16, wherein said method are to be used to handle the skin exogenous injury.

22, the method for claim 17, wherein said method are to be used to handle the skin exogenous injury.

23, the method for claim 18, wherein said method are to be used to handle the skin exogenous injury.

24, the method for claim 19, wherein said method are to be used to handle the skin exogenous injury.

25, the method for claim 20, wherein said method are to be used to handle the skin exogenous injury.

26, the method for claim 16, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

27, the method for claim 17, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

28, the method for claim 18, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

29, the method for claim 19, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

30, the method for claim 20, wherein said soybean prod have the mercaptan retentive activity at least about 75%.

31, the compositions of claim 1, wherein said soybean prod is handled with gamma ray radiation.

32, the method for claim 16, wherein said soybean prod is handled with gamma ray radiation.