CN102014930A

CN102014930A - Compositions comprising ARNOX-inhibitors for the inhibition of reactive oxygen species

Info

Publication number: CN102014930A
Application number: CN2008801290521A
Authority: CN
Inventors: D·克恩; C·梅多斯
Original assignee: Nu Skin International Inc; Nox Technologies Inc
Current assignee: Nu Skin International Inc; Nox Technologies Inc
Priority date: 2008-03-28
Filing date: 2008-03-28
Publication date: 2011-04-13
Also published as: AU2008353441A1; IL208324A0; JP2011515466A; MX2010010668A; EA201001558A1; EP2280714A1; WO2009120214A1; ZA201007247B; KR20110000745A; AU2008353441A2; CA2719833A1

Abstract

The invention relates to agents for sequestering serum aging factors, and methods for using the same. More particularly, the invention relates to agents termed herein 'Naractin' to denote any one of several naturally-occurring arNOX inhibitors either present in N. tazetta powder or capable of augmenting N. tazetta powder to an inhibitory level comparable to that of the fresh N. tazetta extracts, and to methods for using 'Naractins' to prevent or treat disorders and complications of disorders resulting from cell damage caused by an aging-related isoform of NADH oxidase (arNOX). In one exemplary embodiment the agents of the invention comprise at least one naturally occurring Naractin. Such naturally occurring naratins are also capable of augmenting the anti-arNOX effect of other naturally occurring arNOX inhibitory agents.

Description

Be used to suppress the compositions that comprises the ARNOX-inhibitor of reactive oxygen species

Invention field

The present invention relates to be used for the natural extracts of isolate serum senescence-factor, this extract can be by inside or topical.More specifically, the present invention relates to be used to improve looks and go up reagent and compositions and its method that suppresses or improve old and feeble relevant oxidation as skin-protection product.

Background of invention

Plasma membrane nadh oxidase (NOX) is the cell cortex protein with uniqueness of hydroquinone (NADH) oxidase and disulfide protein-mercaptan exchange activity, and it is reacted to hormone and somatomedin usually.NOX (or CLOX) is the growth associated protein matter family relevant with senile cell.Insensitive and the drug responsiveness form to hormone of the NOX of called after tNOX is addressed, and this form is special to cancerous cell.For example, referring to U.S. Patent number 5,605,810, be incorporated herein it all as a reference.

The relevant isotype of the aging of nadh oxidase (arNOX) is a member of this protein families.The circulation form of arNOX significantly increases in human serum and individual lymphocyte, and especially the age is between 30 to 65.The characteristics of arNOX protein uniqueness are to produce the ability of super oxygen base, and it can significantly help old and feeble relevant variation, comprise that atheroma forms and other action at distance aging phenomenons.State before the activity of arNOX in senile cell and the serum.Referring to as the open application number WO 00/57871 of PCT, be incorporated herein it all as a reference.

The model of this arNOX effect is consistent with mitochondrion old and feeble theoretical (Mitrochondrial Theory of Aging), this theory is thought in the aging course, the reactive oxygen species that increases in the mitochondrion causes Mitochondrial DNA Mutation, and lesion wire plastochondria component, causes aging.The old and feeble theoretical accumulation that proposes spontaneous somatic mutation in the mitochondrial DNA (mtDNA) of mitochondrion has caused mistake (Manczak M et al., the J Neurochem.2005 Feb of mtDNA encoded polypeptides chain; 92 (3): 494-504).These mistakes that are created in the mtDNA encoded polypeptides chain are at random, and transmit randomly during mitochondrion and cell division.The result of these changes is deficiency oxidative phosphorylation.The respiratory chain defective may with oxidative stress relevant (Ozawa, 1995, the Biochim.Biophys.Acta 1271:177-189 of the increase of amplifying former damage; And Lenaz, 1998, Biochim.Biophys.Acta 1366:53-67).Therefore this shows that although be present in the body with unusual trace, the mitochondrial DNA of variation may be the main generation body of oxidative stress.

When the accumulation of mtDNA somatic mutation causes the deficiency oxidative phosphorylation, plasma membrane oxidoreductase (PMOR) system has been proposed, increase survival rate (de Grey, 1997, the BioEssays 19:161-166 of mitochondrial defects cell by the regeneration of pyridine oxide nucleotide; De Grey, 1998, Anti-Aging Med.1:53-66; Yoneda etc., 1995, Biochem.Biophys.Res.Comm, 209:723-729; Schon etc., 1996, Cellular Aging and Cell Death, Wiley and Sons, New York, pp.19-34; Ozawa, 1997, Physiol.Rev.77:425-464; And Lenaz, 1998, BioFactors 8:195-204).The damage accumulation of mtDNA is reacted---oxidation as lipid in the low density lipoprotein, LDL (LDLs) changes with the tremulous pulse of following---model that is connected with born of the same parents outward, proposed (Larm etc., 1994, Biol.Chem.269:30097-30100 first with rho ° of cell; Lawen etc., 1994, Mol.Aspects.Med.15:s13-s27; De Grey, 1997, BioEssays 19:161-166; With de Grey, 1998, Anti-Aging Med.1:53-66).With the people's cell that transforms in cultivating similarly study (Vaillant etc., 1996, Bioenerg.Biomemb.28:531-540).

Plasma membrane oxidoreductase (PMOR) is crossed under the situation about expressing, and electronics is transferred to external receptors by the electron transport chain that limits from NADH, causes cell surface reactive oxygen species (ROS) to produce.The ROS that produces of these cell surfaces can will be derived from that mitochondrial old and feeble cascade reaction is delivered to flanking cell and as the blood circulation component of low density lipoprotein, LDL then.Referring to the open application number WO 00/57871 of PCT, be incorporated herein it all as a reference.

Therefore, need find such reagent: it reduces the ability that arNOX produces reactive oxygen species (ROS), reduces or treatment is produced as the purpose of physiology's situation of the oxidation of lipid in the low density lipoprotein, LDL (LDLs) and the tremulous pulse variation followed to reach.The arNOX activity that has shown senile cell is subjected to the natural inhibition that has agent as ubiquinone (ubiquinone).Referring to PCT open application number WO 00/57871, WO 01/72318 and WO 01/72319, be incorporated herein its full content as a reference.But owing to following reason, the application of ubiquinone is not exclusively satisfactory: its price is high, and its easy oxidation loses it and renders a service, and comprise the prepared product of ubiquinone must be by special parcel to prevent loss function.Therefore, although some can suppress the active reagent of arNOX and method exists at present, challenge also still exists.Thus, with suppress that arNOX and avirulence and naturally occurring reagent and technology increase or even replace before disclosed reagent and technology will be progress in this area.

Particularly skin is subjected to the reactive oxygen species damage easily.Skin is made up of two main layers.Horny layer or epidermis are the upper stratas, for skin forms flowing of overcoat and control water and material turnover skin.Corium is the lower floor of skin, for skin provides dynamics, elasticity and thickness.The main cell type of corium is a fibroblast, and it is responsible for the synthetic and secretion as all dermal matrix components of collagen protein, elastin laminin and mucopolysaccharide.Collagen protein provides dynamics, and elastin laminin provides elasticity, and mucopolysaccharide provides the humidity and the plumpness of skin.

Except being damaged by reactive oxygen species, skin suffers various damage stresser.Skin may be by multiple factor damage or injury in the environment.Some are spontaneous, as sun UV radiation, wind even mechanical wounding, as cutting, scrape the similar injury of scrape along.In addition, the also daily generation of artificial injury.These injuries comprise the use of soap, emulsifier based cosmetics, hot water, organic solvent, air-conditioning and central heating.In addition, other injuries to skin can come from dermatosis or normal aging course (chronic aging) or its part, and it can be exposed to various outside stresser (for example light decay is old) by skin and accelerate.

Everyone skin is old and feeble in time.Yet people's life is longer in the modern society, and normal aging effect has an opportunity to obtain accumulation.This effect may be on improving looks fully, as the increase of wrinkle or " senile plaque "; Or it may be influential to health, as be exposed to the generation of the skin carcinoma that UV light causes.Along with people's ageing, thinning of skin, the connective tissue of skin, collagen protein and elastin laminin change, and cause skin to lose hardness and become dry.And the sweat gland of skin and oil droplet activity reduce, thereby cause skin to lose humidity and dry.Further, it is fragile more that skin heart becomes, thereby it breaks and leaks in skin.

The symptom of old and feeble skin comprises xerosis cutis, pruritus, attenuation or thickens, wrinkle and microgroove, often refers to the pigmentation district of chloasma hepaticum and the zone under angiorrhexis (telangiectasis) skin.

The beauty treatment and the medical product that need " defying age ", its treatment or delay actual aging and the visible signs of air slaking skin is as wrinkle, microgroove, sagging, pigmentation and senile plaque.But great majority beauty treatment or medical product do not solve the basic of these symptoms, for example derive from the generation and the accumulation of the relevant free radical of arNOX of ROS.Therefore, need effective natural skin treatment and protection component and application process thereof.

Summary of the invention

The present invention relates to be used for isolating the reagent and the application process thereof of (or chelating) serum senescence-factor.More specifically, the present invention relates to be called the reagent of " Naractin " herein, expression is present in maybe can strengthen narcissus powder any one to several natural arNOX of the existence inhibitor of the inhibition level of the bright narcissus extract that is equal in narcissus (N.tazetta) powder; The disease that produces and the method for disease complication with relating to the cell injury that application " Naractins " prevention or the treatment relevant isotype of aging by nadh oxidase (arNOX) causes.In an illustrative embodiments, reagent of the present invention comprises at least a naturally occurring Naractin.This naturally occurring naratins also can strengthen the anti-arNOX effect of other the natural arNOX of existence inhibitor.

Therefore, in an illustrative embodiments, the present invention includes the topical composition that is used to improve aging effect, said composition comprises at least a arNOX inhibitor of effective dose.According to this illustrative embodiments, the arNOX inhibitor is naractin, and naractin effectively reduces aging effect.Some according to an illustrative embodiment of the invention in, particularly extract and/or purification naractin aspergillosis (Aspergillus), Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis (spurge), filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi) (lithospermum), Nicotiana tabacum L. or the Herba Visci from narcissus, willow, corn, crepis, willow, Caulis Viburni Dilatati, mycete.

In other different embodiments, naractin is Salicylate and derivant thereof.In some illustrative embodiments, Salicylate is that Salicylate is salicin, salicylic acid, salicylhydroxamic acid salt (salicyl hydroxamate), its derivant or its combination.In different illustrative embodiments, naractin derives from Chinese Radix Alangii (Alangium chinense), three split Radix Alangii (A.platanifolium), Japan Radix Alangii (A.premnifolium), aspergillus niger (Aspergillus niger), Betula platyphylla Suk. (Betula alba), Radix Stellariae (Bupleurum falcatum), Herba Catharanthi Rosei (Catharanthus roseus), korean willow (Chosenia bracteosa), Colchicum autumnale (Colchicum autumnale), crepis (Crepis foetida), C.rhoeadifolia, hair leaf Flos Daturae (Datura inoxia), Australian eggplant (Duboisia myoporoides), Eleutherococcus setchuensis, Euphorbia salicifolia, Filipendula ulmaria, Foeniculum vulgare, Fructus Gardeniae (Gardenia jasminoides), Radix Arnebiae (Radix Lithospermi) (lithospermum erythrorhizon), safflower tobacco (Nicotiana tabacum), white poplar (Populus alba), face cream poplar (P.balsamifera), Populus davidiana (P.davidiana), Populus deltoides Bartr.cv. (P.deltoides), Populus euphratica (P.euphratica), canine tooth poplar (P.grandidentata), P.heterophylla, Populus lasiocarpa Oliv. (P.lasiocarpa), the Liao Dynasty poplar (P.maximowiczii), Lombardy poplar (P.nigra), P.sieboldii, PopulussimoniiCarr. (P.simonii), Cathay poplar (P.tacamahaca), Cortex Populi Tomentosae (P.tomentosa), Populus tremula (P.tremula), Populus tremuloides (P.tremuloides), comospore poplar (P.trichocarpa), point leaf willow (Salix acutifolia), Salix alba (S.alba), S.americana, arctic willow (S.arctica), ear willow (S.aurita), Salix babylonica L. (S.babylonica), S.basfordiana, Europe purple willow (S.caesia), the U.S. willow (S.calodendron) of Rutaceae, round end willow (S.capitata), Salix caprea (S.caprea), gland willow (S.chaenomeloides), ash willow (S.cinerea), winter daphne willow (S.daphnoides), salix fragilis (S.fragilis), S.geminata, Salix gracilistyla (S.gracilis), Salix gracilistyla (S.gracilistyla), S.gracilistyloides, S.gymnolepis, halberd willow (S.hastata), dwarf willow (S.herbacea), S.incana, point leaf purple willow (S.koriyanagi), S.lapponum, S.lasiandra, S.lasiolepis, dry land willow (S.matsudana), S.myrsinifolia, S.nigricans, S.orestera, bay willow (S.pentandra), S.pentandroides, S.petiolaris, Salix phylicifolia (S.phylicifolia), purple osier (S.purpurea), creeping willow (S.repens), S.rubra, Artemisia willow (S.schwerinii), Scouler willow (Salix scouleriana) (S.scouleriana), S.smithiana, Zhunger Basin willow (S.songarica), S.species, S.stipularis, four sub-willows (S.tetrasperma), S.tremuloides, three stamen willows (S.triandra), Artemisia willow (S.viminalis), Toisusu urbaniana, the east of Sichuan Caulis Viburni Dilatati (Viburnum henryi), viburnum prunifolium (V.prunifolium), wrinkle Ye Caulis Viburni Dilatati (V.rhytidophyllum) or Herba Visci (Viscum album).

In other different embodiments, naractin is Salicylate and derivant thereof.In some illustrative embodiments, Salicylate is salicin, salicylic acid, salicylhydroxamic acid salt, its derivant or its combination.

In other different embodiments, said composition further comprises beauty treatment or pharmaceutically acceptable carrier.In some illustrative embodiments, naractin inhibitor and other derive from the arNOX inhibitor coexistence in natural existence source, and this source includes but not limited to Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle (coleus), Herba Rosmarini Officinalis (rosemary), Flos Nelumbinis, Carlina acaulis (artichoke), extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae (Oenothera biennis), astaxanthin, tangerine, Fructus Schisandrae Chinensis (Schisandra chinensis), Radix Ophiopogonis (Lonicera), Semen Fagopyri Esculenti (Fagopyrum), Radix Dauci Sativae or Fructus Canarii albi.In different illustrative embodiments, naractin strengthens the effect of additional arNOX inhibitor.

Skilled person in the art will appreciate that arNOX composite inhibiting described herein can be with any convenient mode administration.In some illustrative embodiments, such form of medication comprises unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen.In different illustrative embodiments, the aging effect that is improved by the present invention includes but not limited to microgroove, wrinkle, pigmentation, dewaters, follows the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.In these and other illustrative embodiments of the present invention, provide concentration at about 5 μ g/ml to the arNOX inhibitor between the about 500 μ g/ml.

In also having other illustrative embodiments, the present invention includes inhibition is produced to improve the method for aging effect by the reactive oxygen species that the old and feeble relevant isotype of nadh oxidase causes, comprise: the compositions for the treatment of effective dose to its required patient, said composition comprises in salicin, salicylic acid, the salicylhydroxamic acid salt at least a, thereby suppressed the generation of the reactive oxygen species that caused by the old and feeble relevant isotype of nadh oxidase, wherein aging effect improves.In different illustrative embodiments, this method further comprises extract at least a in the following material or purified extract: Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae, astaxanthin, tangerine, Fructus Schisandrae Chinensis, Radix Ophiopogonis, Semen Fagopyri Esculenti, Radix Dauci Sativae, narcissus, Fructus Canarii albi, willow, Herba bromi japonici or corn.In according to different illustrative embodiments of the present invention, compositions is as unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen are used.In these illustrative embodiments, aging effect includes but not limited to microgroove, wrinkle, pigmentation, dewaters, follows the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

In also having other illustrative embodiments, the present invention includes the beauty method that improves aging effect, comprise that said composition comprises to the dermal administration cosmetic composition: be enough to suppress the effective dose naractin of arNOX, the aging effect of wherein at least a arNOX mediation is suppressed.In these embodiments, naractin is the Salicylate or derivatives thereof.In different illustrative embodiments, Salicylate is salicin, salicylhydroxamic acid salt or salicylic acid.In other different embodiments according to the method for the invention, cosmetic composition further comprises plant extract, comprising: Radix Dauci Sativae extract, Fructus Canrii Albi extract, cauliflower extract, Lentinus Edodes extract, flame nettle extract, Herba Rosmarini Officinalis extract, Flos Nelumbinis extract, Carlina acaulis extract, extra large rose extract, Citrus reticulata Blanco extract, Radix Oenotherae erythrosepalae extract, tangerine extract, Fructus Schisandrae Chinensis extrat, Radix Ophiopogonis extract, Semen Fagopyri Esculenti extract, willow extract, corn, Herba bromi japonici or narcissus extract.In these illustrative embodiments, naractin provides in the lump with the beauty treatment acceptable carrier.

In different illustrative embodiments, by the aging effect that improves according to method of the present invention comprise microgroove, wrinkle, pigmentation, dewater, follow the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

In different embodiments according to the method for the invention, at least one day applied once naractin.In some illustrative embodiments, in beautification product, provide naractin with the concentration of about 5 μ g/ml between about 500 μ g/ml.In these illustrative embodiments, will be with unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen give according to cosmetic composition of the present invention.

In with good grounds other embodiments of the present invention also, the present invention includes test kit, this test kit is used to use to improving the useful cosmetics of aging effect, and this test kit comprises: at least a naractin; And operation instruction.In some illustrative embodiments, this test kit further comprises such beautification product, and it is suitable for suppressing as at least a arNOX the carrier of plant extract.

These and other features of the present invention and advantage will be set forth and maybe will be become more fully high-visible in following description and appended claims.Can realize and obtain this feature and advantage by equipment and the combination of specifically noting in the appended claims.In addition, feature of the present invention and advantage can arrive by practice knowledge of the present invention, or high-visible by description, as mentioned below.

The accompanying drawing summary

To describe different illustrative embodiments in detail with reference to figure below according to compositions of the present invention and method, wherein:

Part inhibition (middle graph) after Fig. 1 a shows the activity figure (BL=baseline) of arNOX isotype in 72 years old (y/o) male saliva sample and adds the narcissus powder extracts.Right figure shows that adding salicylhydroxamic acid salt (SHAM) (Naractin 1) composition almost completely activity behind the narcissus powder suppresses.Except substituting SHAM, Fig. 1 b is identical with Fig. 1 a, and the inhibition effect of narcissus powder strengthens with salicin (Naractin 3).Naractins and ferrum form the red complex of the spectral quality with similar redness, the part and the extract that comprises the arNOX mortifier of this property representation narcissus bulb part as shown in Figure 2.

Fig. 2 a-f shows narcissus bulb zone and comprises the extract of the active mortifier of arNOX.Mortifier is not present in the bulb part and/or extract that color do not become redness or pink colour.Fig. 2 a is bulb figure, and this bulb is divided into the zone of being made up of leaf district and stem district.Fig. 2 b shows that the red compound relevant with the arNOX inhibition is positioned the photo in the stem district.Fig. 2 c shows the different bulbs zone that extract forms, and has different red formation levels in the zones of different, shown in Fig. 2 d, 2e and 2f.Color forms rapidly.The bulb that extract does not suppress the active narcissus kind of arNOX can not form redness yet.

After Fig. 3 shows interpolation iron chloride, the similar redness of corn dimension body (steeles) (the central hadrome of root) vasculature.Produce red material and be naturally occurring hydroxamate (hydroxamate, hydroxamate).

Fig. 4 shows the active inhibition of arNOX in the saliva (72 years old, the male) in baseline (BL) figure, and shows the inhibition that is caused by the made homogenate of corn dimension body by adding.Corn sheath (surrounding the tissue of dimension body) is not painted because of the interpolation of ferrum, does not suppress the arNOX activity yet.

The thin-layer chromatographic analysis example of narcissus bulb methanolic extract (B) is compared in Fig. 5 demonstration with commodity narcissus powder (A).The flat pannel display Ultraluminescence of blue (UV) background.The flat board of light background (BB) is dyeed by Berlin blue.The TLC system is a dichloromethane: methanol: NH ₄OH (10: 1: 0.2).In the sample that laboratory extracts, red component---it is last, but clearly separates with the material of initial point, and the existence of hint hydroxamate---and much obvious in laboratory extracts in the sample than the commodity powder.This for methanol (Fig. 5 a) and water (Fig. 5 b) extract all apparent.Also as seen the hydroxamate of inferring, but is extracting far below laboratory on the level of sample in the commodity powder, and to suppress active level relevant with arNOX once more.

After Fig. 6 showed that adding iron chloride forms red ferrum hydroxamate, (SHAM) compared with known hydroxamate, the spectrum analysis of narcissus bulb and corn dimension body and sheath extract.It is red that the arNOX of narcissus bulb suppresses extract, and presents maximum absorbance about 550nm.It is colourless not suppress active daffadowndilly (N.pseudo narcissus) bulb extract, and does not show absorbance at 550nm.Do not add iron chloride (FeCl ₂) time, Zea mays root dimension body (activity) and corn root sheath (non-activity) part are not painted to a great extent.But there is (+FeCl in iron chloride ₂) time, corn dimension body extract occurs red, have maximum absorbance at about 550nm, and the corn sheath does not have maximum absorbance at 550nm.Salicylhydroxamic acid salt (SHAM), a kind of commercially available stable hydroxamate also is given in the redness that about 550nm has maximum absorbance.Negative control is not painted, and does not show absorbance at 550nm.

Fig. 7 is the active figure of the arNOX of ferricytochrome c, and the arNOX activity is the function of SHAM diluent (log).This chart understands that (Sigma-Aldrich, St.Louis MO) suppress the active dose dependent of saliva (72 years old, the male) arNOX commercial obtainable SHAM goods.Substantially fully suppress at 11mg/ml.

Fig. 8 is the active figure of the arNOX of ferricytochrome c, and the arNOX activity is the function of salicylic acid diluent (log).This chart is understood the active dose dependent inhibition to saliva (72 years old, the male) arNOX, and the arNOX activity is the function of salicylic acid concentration.Salicylic acid suppresses about 50% under 1mg/ml concentration.

Fig. 9 is the active figure of the arNOX of ferricytochrome c, and the arNOX activity is the function of salicin diluent (log).This chart is understood the active dose dependent inhibition to saliva (72 years old, the male) arNOX, and the arNOX activity is the function of salicin concentration.Reach the active inhibition of maximum arNOX in 1mg/ml concentration.

Figure 10 a and 10b are the figure that shows the arNOX inhibition of multiple " Naractin " combination.Figure 10 a shows the salicin potentiation that the 1mg/ml salicin exists the arNOX of the mixture of following 4mg/ml Fructus Schisandrae Chinensis (Schizandra) powder and narcissus extract (20 μ l) to suppress.Figure 10 b shows that the arNOX that the 1mg/ml salicin exists the mixture of following 4mg/ml Chinese Magnolivine Fruit and 1mg/ml narcissus powder to cause suppresses.The detection mixture that comprises saliva (72 years old, the male) to total amount 3ml adds 60 μ l mixture, and this saliva is as the active source of arNOX.

Detailed Description Of The Invention

The present invention relates to reagent and application process thereof for the isolate serum senescence-factor. More specifically, the present invention relates to be called the reagent of " Naractin " herein, represent in several natural arNOX of existence inhibitor wherein any one, it is present in maybe can strengthen the narcissus powder to the inhibition level of the bright narcissus extract that is equal in the narcissus powder; The disease that causes and the method for disease complication with relating to the cellular damage that application " Naractins " prevention or the treatment relevant isotype of aging by nadh oxidase (arNOX) causes. In an illustrative embodiments, reagent of the present invention comprises at least a naturally occurring Naractin. This naturally occurring naratins also can strengthen the anti-arNOX effect of other the natural arNOX of existence inhibitor.

Term " Naractin " this paper be used for expression be present in the narcissus powder (commercial can be from for example Xian Aojing Science and Technology Developing CO., LTD, Xi'an, Shaanxi, China obtains) maybe can strengthen to several natural arNOX of the existence inhibitor of the inhibition level of the bright narcissus extract that can be equal to any one of narcissus powder. The inventor confirmed at present three kinds of known having "Naractin" active chemical pure material. Although should be noted that further this paper differentiates " Naractins " come out from the narcissus extract, it may be present in other natural origins, such as willow, corn and oat, certainly, the present invention includes Naractins, no matter how it originates.

As used herein, term " cosmetics " refers to be intended to clean, beautify, improve for body the material of glamour or change outward appearance.

As used herein, term " extract " refers to by dipping in solvent or soaks material and remove the resulting solution of active component. Solvent can be any suitable solvent, includes but not limited to alcohol, water or analog. In some instances, extract can concentrate or solvent can evaporate, and active component can suspend or be dissolved in the different solvents. As described herein, make the narcissus extract by the stem district that homogenizes from bulb separate stem district and 3ml distilled water.

As used herein; term " disease " refers to any state that live animal or plant body or the normal function of its part weaken; comprise any disease (ailment; disease, illness), clinical state, pathological state, the state that dies down, unsound state and physiological status unusually any or that do not expect.

As used herein, term " reactive oxygen species " refers to cause the formation (such as peroxide or hydroxyl radical free radical) of free radical from the oxygen derivative of oxygen metabolism or free electron transfer.

As used herein, term " antioxidant " refers to the active of elimination activity oxygen species or the compound of the cellular damage that suppresses to be caused by described active matter.

As used herein, term " pharmaceutically acceptable carrier " refers to such mounting medium: its biologically active that does not hinder active component is renderd a service, and has chemical inertness, and to the patient's avirulence with its administration.

As used herein, term " pharmacy acceptable derivates " refers to and the corresponding any homologue of the described preparation of the application, analog or fragment that it is antioxidation activity, and to the relative avirulence of subject.

Term " therapeutic agent " refers to disease that any auxiliary prevention or treatment are caused by reactive oxygen species or molecule, compound or the therapeutic agent of disease complication, preferred anti-oxidant.

Term " reagent of isolation arNOX " refers to any such molecule, compound or therapeutic agent: itself and arNOX interact, thereby reduce the reaction of arNOX and other materials and suppress the ability that arNOX produces reactive oxygen species.

Term used herein " salicylate " refers to salt, conjugate or the derivative of salicin, no matter derives from natural existence source or again synthetic. These salicylates comprise, such as salicylic acid, SHAM and salicin itself, its derivative, salt and conjugate.

During its research, the inventor finds that some narcissus extract article of commerce comprises and has the active activating agent of anti-arNOX.By observing prompting the present invention, IBR-DORMIN

In fact some samples of narcissus bulb extract (Israeli Biotechnology Research, Ltd., Israel) do not have the activity of inhibition to arNOX, and other multiple samples of this manufacturer have strong rejection capability.The inventor guesses that arNOX inhibitor instability or arNOX inhibitor under specific antiseptic disappearance incur loss at narcissus extract purification process.Yet should be noted in the discussion above that trade mark IBR-Dormin The commercially available narcissus powder of being sold is through special processing, and the narcissus bulb is in hibernation.Principle is to produce a large amount of unacknowledged chemical compounds at the trophophase bulb, is called " dormin ".Further conjecture this " dormin " causes the resting state of cell or tissue.Therefore, from data show of the present disclosure, narcissus " dormin " is not " Naractins " identical reagent of being confirmed with this paper.

In order to check these probabilities, the inventor obtains pure white on a small quantity (narcissus) bulb.Water and ethanol extraction suppress arNOX.With do not have active N.pseudo-narcissus (daffadowndilly) to a great extent and N.jonquilla (daffodil) compares.Active and relevant by adding ferric iron enhanced red colored---characteristic of naturally occurring hydroxamate---, this hydroxamate is found in corn and avenaceous rice shoot (data not shown), and it also suppresses arNOX.Hydroxamate has such chemical property: expection has the arNOX of benefiting and suppresses.The hydroxamate of estimating the easiest acquisition is salicylhydroxamic acid salt (Naractin 1), is also referred to as SHAM, known plant alternating oxidation enzyme inhibitor.Detect SHAM, and find that it strengthens the arNOX inhibitory reaction of narcissus powder.Along with the narcissus powder strengthens with low-level SHAM, obtain being higher than 90% inhibition level.Though the narcissus bulb is not enough to finish the sign of active component, and usefulness narcissus powder (extract) (Xi ' an Aojing Science and Technology Developing Co., LTD., Xi'an, Shaanxi, China; 20: 1 obtainable bulbs and flower extract) proceed research.The narcissus powder presents really and contains low-level naturally occurring hydroxamate, but on the level far below the made extract of narcissus bulb.

What the problem that causes was a hydroxamate or salicylic acid part for the arNOX inhibitor is active most important.The result shows that the two is all important.Salicylic acid (Naractin 2) itself is very effective arNOX inhibitor, and when reacting with ferric iron, it also takes on a red color.Salicylate or aspirin do not suppress arNOX.Detect the water extract of bark of willow---natural origin of Salicylate---, find that also it suppresses arNOX.Main Salicylate---salicin---glucosides of bark of willow, also effective as inhibitor (Naractin 3).

Therefore, in an illustrative embodiments, the present invention includes the topical composition that is used to improve aging effect, comprise at least a arNOX inhibitor of effective dose.According to this illustrative embodiments, the arNOX inhibitor is naractin, and naractin effectively reduces aging effect.Some according to an illustrative embodiment of the invention in, from narcissus, willow, corn, crepis, willow, Caulis Viburni Dilatati, the mycete naractin that particularly purifies aspergillosis (Aspergillus), Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis, filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi), Nicotiana tabacum L. or the Herba Visci.

In other different embodiments, naractin is Salicylate and derivant thereof.In some illustrative embodiments, Salicylate is that Salicylate is salicin, salicylic acid, salicylhydroxamic acid salt, its derivant or combination.In different illustrative embodiments, naractin derives from Chinese Radix Alangii (Alangium chinense), three split Radix Alangii (A.platanifolium), Japan Radix Alangii (A.premnifolium), aspergillus niger (Aspergillus niger), Betula platyphylla Suk. (Betula alba), Radix Stellariae (Bupleurum falcatum), Herba Catharanthi Rosei (Catharanthus roseus), korean willow (Chosenia bracteosa), Colchicum autumnale (Colchicum autumnale), crepis (Crepis foetida), C.rhoeadifolia, hair leaf Flos Daturae (Datura inoxia), Australian eggplant (Duboisia myoporoides), Eleutherococcus setchuensis, Euphorbia salicifolia, Filipendula ulmaria, Foeniculum vulgare, Fructus Gardeniae (Gardeniajasminoides), Radix Arnebiae (Radix Lithospermi) (Lithospermum erythrorhizon), safflower tobacco (Nicotiana tabacum), white poplar (Populus alba), face cream poplar (P.balsamifera), Populus davidiana (P.davidiana), Populus deltoides Bartr.cv. (P.deltoides), Populus euphratica (P.euphratica), canine tooth poplar (P.grandidentata), P.heterophylla, Populus lasiocarpa Oliv. (P.lasiocarpa), the Liao Dynasty poplar (P.maximowiczii), Lombardy poplar (P.nigra), (P.sieboldii), PopulussimoniiCarr. (P.simonii), Cathay poplar (P.tacamahaca), Cortex Populi Tomentosae (P.tomentosa), Populus tremula (P.tremula), Populus tremuloides (P.tremuloides), comospore poplar (P.trichocarpa), point leaf willow (Salix acutifolia), Salix alba (S.alba), S.americana, arctic willow (S.arctica), ear willow (S.aurita), Salix babylonica L. (S.babylonica), S.basfordiana, Europe purple willow (S.caesia), the U.S. willow (S.calodendron) of Rutaceae, round end willow (S.capitata), Salix caprea (S.caprea), gland willow (S.chaenomeloides), ash willow (S.cinerea), winter daphne willow (S.daphnoides), salix fragilis (S.fragilis), S.geminata, Salix gracilistyla (S.gracilis), Salix gracilistyla (S.gracilistyla), S.gracilistyloides, S.gymnolepis, halberd willow (S.hastata), dwarf willow (S.herbacea), S.incana, point leaf purple willow (S.koriyanagi), S.lapponum, S.lasiandra, S.lasiolepis, dry land willow (S.matsudana), S.myrsinifolia, S.nigricans, S.orestera, bay willow (S.pentandra), S.pentandroides, S.petiolaris, Salix phylicifolia (S.phylicifolia), purple osier (S.purpurea), creeping willow (S.repens), S.rubra, Artemisia willow (S.schwerinii), Scouler willow (Salix scouleriana) (S.scouleriana), S.smithiana, Zhunger Basin willow (S.songarica), S.species, S.stipularis, four sub-willows (S.tetrasperma), S.tremuloides, three stamen willows (S.triandra), Artemisia willow (S.viminalis), Toisusu urbaniana, the east of Sichuan Caulis Viburni Dilatati (Viburnum henryi), viburnum prunifolium (V.prunifolium), wrinkle Ye Caulis Viburni Dilatati (V.rhytidophyllum) or Herba Visci (Viscum album).

In other different embodiments, naractin is Salicylate and derivant thereof.In some illustrative embodiments, Salicylate is salicin, salicylic acid, salicylhydroxamic acid salt, its derivant or combination.

In other different embodiments, compositions further comprises beauty treatment or pharmaceutically acceptable carrier.In some illustrative embodiments, naractin inhibitor and other arNOX inhibitor coexistence that derives from natural origin, this source include but not limited to Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae, astaxanthin, tangerine, Fructus Schisandrae Chinensis, Radix Ophiopogonis, Semen Fagopyri Esculenti, Radix Dauci Sativae or Fructus Canarii albi.In different illustrative embodiments, naractin strengthens the effect of additional arNOX inhibitor.

Those of skill in the art will recognize that can be with arNOX composite inhibiting as herein described with any easy way administration.In some illustrative embodiments, these form of medication comprise unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen.In different illustrative embodiments, the aging effect that is improved by the present invention includes but not limited to microgroove, wrinkle, pigmentation, dewaters, follows the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.In these and other illustrative embodiments of the present invention, provide arNOX inhibitor with the concentration of about 5 μ g/ml between about 500 μ g/ml.

In also having other illustrative embodiments, the present invention includes inhibition is produced to improve the method for aging effect by the reactive oxygen species that the old and feeble relevant isotype of nadh oxidase causes, comprise: the compositions for the treatment of effective dose to its required patient, said composition comprises in salicin, salicylic acid, the salicylhydroxamic acid salt at least a, suppress thus to be produced by the reactive oxygen species that the old and feeble relevant isotype of nadh oxidase causes, wherein aging effect improves.In different illustrative embodiments, this method further comprises extract at least a in the following material or purified extract: Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae, astaxanthin, tangerine, Fructus Schisandrae Chinensis, Radix Ophiopogonis, Semen Fagopyri Esculenti, Radix Dauci Sativae, narcissus, Fructus Canarii albi, willow, Herba bromi japonici, corn, crepis, willow, Caulis Viburni Dilatati, mycete particularly aspergillosis (Aspergillus), Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis, filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi), Nicotiana tabacum L. or Herba Visci.In according to different illustrative embodiments of the present invention, compositions is as unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo, ingredients or sunscreen are used.In these illustrative embodiments, aging effect includes but not limited to microgroove, wrinkle, pigmentation, dewaters, follows the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

In also having other illustrative embodiments, the present invention includes the beauty method that improves aging effect, comprise that said composition comprises to the dermal administration cosmetic composition: effective dose is enough to suppress the naractin of arNOX, and the aging effect of wherein at least a arNOX mediation is suppressed.In these embodiments, naractin is Salicylate and derivant thereof.In different illustrative embodiments, Salicylate is salicin, salicylhydroxamic acid salt or salicylic acid.In other different embodiments according to the method for the invention, cosmetic composition further comprises plant extract, comprising: Radix Dauci Sativae extract, Fructus Canrii Albi extract, cauliflower extract, Lentinus Edodes extract, the flame nettle extract, Herba Rosmarini Officinalis extract, Flos Nelumbinis extract, Carlina acaulis extract, the sea rose extract, Citrus reticulata Blanco extract, Radix Oenotherae erythrosepalae extract, the tangerine extract, Fructus Schisandrae Chinensis extrat, Radix Ophiopogonis extract, Semen Fagopyri Esculenti extract, the willow extract, corn, crepis, willow Caulis Viburni Dilatati, mycete is aspergillosis (Aspergillus) particularly, Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis, filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi), Nicotiana tabacum L., Herba Visci, Herba bromi japonici or narcissus extract.In these illustrative embodiments, naractin provides with the beauty treatment acceptable carrier.

In different embodiments according to the method for the invention, at least one day applied once naractin.In some illustrative embodiments, in beautification product, provide naractin with the concentration of about 5 μ g/ml between about 500 μ g/ml.In these illustrative embodiments, will be according to unguentum, emulsion, washing liquid, ingredients, the suspension of cosmetic composition of the present invention with lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen administration.

In with good grounds other embodiments of the present invention also, the present invention includes test kit, this test kit is used to use to improving the useful cosmetics of aging effect, and this test kit comprises: at least a naractin; And operation instruction.In some illustrative embodiments, this test kit further comprises such beautification product: the carrier that is suitable for suppressing as at least a arNOX plant extract.

Antioxidant, cellular component and the target protein of this paper definition are write a Chinese character in simplified form as follows:

Mitochondrial DNA mtDNA

Nicotinamide adenine dinucleotide NADH

Has protein disulfide mercaptan isomerase activity

Cell surface hydroquinone (NADH) oxidase NOX

At the special NOX cNOX of non-cancerous cell

At the special NOX arNOX of senile cell

At the special NOX tNOX of cancerous cell

Low density lipoprotein, LDL LDL

Plasma membrane oxidoreductase chain PMOR

Ubiquinone or ubiquinone CoQ

Auxilliary Q ₁₀CoQ ₁₀

Reactive oxygen species ROS

The present invention

The present invention relates to from narcissus bulb and corn the natural reagent that exists identifying and purify, and can inside or topical.These medicaments suppress arNOX specifically and improve some its old and feeble correlation effects.These reagent can be taked the form of separation agent or plant extract.Further, though the arNOX inhibitor can use separately, it also can be used as and comprises multiple arNOX inhibitor and/or comprise that the compositions of preparation that body is had a chemical compound of other beneficial effects uses.Particularly, the inventor finds that inhibitor has the beneficial effect that strengthens the normal skin care regimen by adding the arNOX inhibitor to cosmetics.

In also having another illustrative embodiments, the present invention includes the cosmetic composition that improves aging effect, said composition comprises at least a arNOX inhibitor of the effective dose of improving looks, and wherein the arNOX inhibitor effectively reduces the skin aging effect.In a kind of form of this illustrative embodiments, the present invention includes the beauty treatment acceptable carrier.In this embodiment, carrier can comprise powder, lubricant, washing liquid, unguentum, liquid and analog.In some illustrative embodiments, the arNOX inhibitor derives from plant.In specific illustrative embodiments, plant be selected from Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae, astaxanthin, tangerine, Fructus Schisandrae Chinensis, Radix Ophiopogonis, Semen Fagopyri Esculenti, Radix Dauci Sativae, narcissus, willow, corn,, corn, crepis, willow, Caulis Viburni Dilatati, mycete particularly aspergillosis (Aspergillus), Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis, filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi), Nicotiana tabacum L., Herba Visci, Herba bromi japonici or Fructus Canarii albi.

Should be understood that, can with according to the described cosmetic composition of this illustrative embodiments with any exemplary approach administration.For example, in some illustrative embodiments, outer or percutaneous of local, oral, intestinal or per rectum are used according to cosmetic composition of the present invention.In some illustrative embodiments, compositions is formulated into unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap or shampoo.

In also having other illustrative embodiments, the present invention includes the beauty method that improves aging effect, comprise the cosmetic composition that comprises effective dose arNOX inhibitor to dermal application, the aging effect of wherein at least a arNOX mediation is suppressed.Some according to an illustrative embodiment of the invention in, the arNOX inhibitor is a plant extract.In other illustrative embodiments, purification arNOX inhibitor from plant extract.In according to different illustrative embodiments of the present invention, the arNOX inhibitor exists with the concentration of about 5 μ g/ml between about 500 μ g/ml.In different illustrative embodiments, the concentration of activating agent exists with the concentration of about 15 to 100 μ g/ml.In some illustrative embodiments, local, oral, intestinal is outer, percutaneous, per rectum or use according to cosmetic composition of the present invention by any effective ways.In some illustrative embodiments, compositions is mixed with unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap or shampoo.

In also having other illustrative embodiments, the present invention includes test kit.In this embodiment, test kit can comprise a certain amount of arNOX inhibitor and operation instruction.In different illustrative embodiments, test kit can further comprise beautification product, thereby can add the arNOX inhibitor to beautification product before use.

Should be understood that, though used a kind of arNOX inhibitor in illustrative embodiments more of the present invention, in other illustrative embodiments, more than a kind of extract or arNOX inhibitor common application.Further, should be understood that, in the different illustrative embodiments of the present invention, can use or give one or more arNOX inhibitor in many ways.As for example topical, with unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; The mode of soap, shampoo or sunscreen and with tea or capsular mode or any other effective means.

Plasma membrane hydroquinone (NADH) oxidase (NOX):

Plasma membrane nadh oxidase (NOX) is the cell cortex protein with uniqueness of hydroquinone (NADH) oxidase and disulfide protein-mercaptan exchange activity, and it is reacted to hormone and somatomedin usually.Called after tNOX is addressed to the insensitive and active drug responsiveness form of hormone, and this form is special to cancerous cell.The evidence that has NOX protein can produce ROS in some cases exists.For example, the ultraviolet of using as oxidative stress source in the cultured cell causes that peroxide produces (Morr é etc., 1999, Biofactors 9:179-187) (referring to U.S. Patent number 5,605,810, being incorporated herein it all as a reference).

Separation and sign are from the arNOX inhibitor of narcissus

By observing some commercial narcissus extract (commercial IBR-DORMIN of can be used as that obtains

, IBR, Inc, Israel obtains) and sample has strong rejection capability to arNOX usually, and other a great differences but goods sample of the same type then do not have, and have promoted guiding series of studies of the present invention.Inventor's hypothesis, it is because some materials are unstable under specific antiseptic disappearance that arNOX suppresses, or the arNOX constituents for suppressing of active product loses from the non-activity goods in the extract purification process.Should be noted in the discussion above that and be called as IBR-DORMIN recently

Product addressed, this product relates to the antiproliferative that derives from plant.U.S. Patent Application Publication No. 20060160702, patent application serial numbers 11/289,156 (' 156 application) referring to Soudant etc.According to ' 156 applications, in plant, find antiproliferative, it can enter resting state (summary) by the inducing plant organ.Therefore, the antiproliferative that proposes to be called " dormin " comes delaying senility to play a role with this by slowing down cell proliferation.The plant extract that has entered rest period by preparation obtains " dormin ".Therefore, have only the extract of dormancy plant can be as " dormin " of this character.

Because the inventor notices different narcissus extracts (the Xian Aojing Science and Technology Developing CO. of commercially available narcissus bulb and flower powder, LTD, Xi'an, China) have arNOX inhibitory action in various degree, so the inventor carries out a series of experiments to identify the arNOX mortifier.In order to check above-mentioned probability, (Brent and Becky ' s bulbs, Gloucester VA) have obtained the bulb of pure white narcissus (N.tazetta) from commercial gardening supplier.With do not have active N.pseudo-narcissus (daffadowndilly) to a great extent and N.jonquilla (daffodil) compares.Active and relevant by adding ferric iron enhanced red colored---characteristic of naturally occurring hydroxamate---.The hydroxamate of estimating the easiest acquisition is salicylhydroxamic acid salt (SHAM) (Naractin 1).Prepare such water and ethanol extraction: it suppresses arNOX and strengthens when in conjunction with low activity narcissus flower powder to suppress (as Fig. 1 a).The similar 1a of Fig. 1 b, the potentiation of demonstration narcissus powder, this narcissus powder derive from bulb (Xian Aojing, LTD) also usefulness salicin (Sigma-Aldrich, St.Louis, MO) (Naractin 3) enhancing of water extraction.Compare with the similar extract that does not have arNOX to suppress active Narcissus pseudo-narcissus (daffadowndilly) and Narcissus jonquilla (daffodil) extract fully.

As shown in Figure 2, the activity of narcissus extract is with relevant by adding ferric iron enhanced red colored---characteristic of naturally occurring hydroxamate---, hydroxamate is such as being found in corn (Fig. 3) and the Herba bromi japonici (not shown) rice shoot those, and it also suppresses arNOX (Fig. 4).By thin-layer chromatographic analysis (Fig. 5), spectral investigation from its ferrum adduct, from Zea mays root (Fig. 6) dimension body (the central vasculature of Zea mays root, wherein hydroxamate is concentrated corresponding to the potential rich hydroxamate stem vasculature of narcissus bulb shown in Figure 2) and by with commercial hydroxamate (the salicylhydroxamic acid salt (SHAM) that obtains, commercial can be available from as Sigma-Aldrich, St Louis, MO.) contrast provides narcissus extract active component to comprise the further evidence of the potentiality of naturally occurring hydroxamate.

Detect active inhibition of arNOX of SHAM, and under concentration 1mg/ml, obtain being higher than active suppress (Fig. 7) of arNOX of 72 years old male's saliva of 90%.Find that also SHAM strengthens active suppress (Fig. 1) of arNOX of the active narcissus powder of part.Commercially available narcissus powder presents really and contains low-level naturally occurring hydroxamate, this hydroxamate may be similar to SHAM, but on far below the hydroxamic acid salt level of from the extract of bright narcissus bulb preparation, being found (Fig. 5 a and 5b).

Whether the arNOX activity is suppressed important problem in order to answer the salicylic acid part of stablizing the salicyl hydroxamate, (Naractin 2 with salicylic acid, commercial can be available from Sigma-Aldrich, St Louis, Mo.) as saliva (72 years old, the male) the active inhibitor of arNOX detects, and finds that it suppresses arNOX (Fig. 8).When reacting with ferric iron, similar to what hydroxamate presented, salicylic acid also takes on a red color.Salicylate and/or aspirin do not suppress arNOX.Detect the water extract of bark of willow---natural origin of Salicylate---, find that it also suppresses arNOX.Also detected salicin (Sigma-Aldrich subsequently, St Louis, Mo) (this paper is appointed as Naractin 3)---the main Salicylate of bark of willow, and find that it has activity (Fig. 9) under the concentration than low 10 times of SHAM (Naractin 1) or salicylic acid (Naractin 2).

Salicin has activity (Fig. 9) during not only as independent reagent, the activity that it also has the arNOX that strengthens other arNOX inhibitor mixed things to suppress, these other arNOX inhibitor are come Chinese freely Chinese Magnolivine Fruit and from the natural origin (Figure 10 a and 10b) of narcissus powder.Three kinds of chemical pure Naractins that identify and detect are (in order to represent wherein any one term of several natural arNOX of existence inhibitor, this the natural arNOX of existence inhibitor or be present in the narcissus powder or can strengthen the inhibition level of narcissus powder to the active bright narcissus extract of tool that is equal to) in, salicin presents and is hopeful most.Further, stable, the water soluble of salicin, nonirritant and relatively cheap.

Usually, the characteristic of senile cell comprises its expression and/or flows out the characteristic of arNOX, and includes but not limited to present one or more following characteristics: old and feeble relevant PMOR system, produce the ability of reactive oxygen species and have the functional defect mitochondrion.One embodiment of the present invention is to utilize reagent to reduce the adverse effect of senile cell.

The symptom of old and feeble skin comprise drying, pruritus, thinning of skin and thicken, wrinkle and microgroove, pigmented area (claiming senile plaque or chloasma hepaticum) and mottled outward appearance.Shown that old and feeble skin collagen reduces and reduces with elasticity.In addition, old and feeble skin has the cracking collagen protein and the cross-linked proteins of recruitment.Shown superoxide radical in these processes.Skin may heal with the longer time when injured.As seen blood vessel is easier sees through attenuation skin, also is because it was expanded with the age.These blood vessels are visible as on the skin damaged blood capillary (telangiectasis) in red dome spline structure (cherry angiomas) or the face.Many people produce aging or actinic purpura, and this purpura is by purple dot or speckle on the skin of a small amount of hemorrhage generation in the skin.Senile skin has less protective effect to sun-damaged, because the protection cell of melanocyte by name reduced with the age.Old and feeble skin also more may produce multiple optimum and precancer growth, as seborrhea and actinic keratosis.Seborrheic keratosis has coarse, brown outward appearance usually, and looks like wart.They are benign.Actinic keratosis is to be subjected to little scale growth on the skin area that the sun is exposed to the sun.They are early stage signs of skin carcinoma.

The topical that the present invention includes natural plant extracts is used, separately or with unguentum, lubricant, washing liquid, gel, lubricant or similar type, to keep skin vitality.Preferred implementation of the present invention comprises the unguentum that comprises arNOX inhibition extract to the patient skin topical administration, to keep and to improve skin vitality.

The beauty therapeutic of skin

The invention provides the compositions that comprises activating agent, said composition is prevented and/or is improved skin injury and conditions associated, is especially caused and the situation relevant with arNOX by aging.Further, the present invention includes the described method for compositions of application.Horny layer is to form top coat and work the skin layer of protecting the skin effect when controlled humidity and material turnover skin flow.When this barrier function damages, the skin influence that sustains damage, thus further impelled presenility.These cause that skin crosses that the damaging action of presenility is that many hope are kept fit, the misgivings place of the individuality of young outward appearance and sense of touch skin.Reactive oxygen species participates in the potential lethal breaking reaction of a plurality of pair cells.Reactive oxygen species partly is responsible for deleterious cell interaction, comprises the ability that fibroblast generates healthy collagen protein and elastin laminin that weakens.In addition, skin is by dermatosis, environmental injury (wind, air-conditioning, central heating) or pass through usual aging process (chronic aging) sustain damage, and this usual aging process can be exposed to the sun (light decay is old) by skin and accelerate.

Preferred implementation of the present invention provides naturally occurring plant promoting agent, is used for the treatment of tissue, especially the arNOX related damage of skin.This activating agent prevention and/or improve skin lesion and conditions associated.In one embodiment of the present invention, the plant product of processing completely cuts off the arNOX activity.In another embodiment of the present invention, the plant product of processing suppresses reactive oxygen species.In another embodiment, reagent of the present invention and method prevention and/or improve skin health.For example, reagent can improve that skin is tight, color and pore is apparent, elasticity, hydration and/or help to reduce that microgroove occurs and old and feeble visible sign.In another illustrative embodiments of the present invention, reagent positive impact health former albumen of natural birth rubber and elastin laminin.In another embodiment, reagent of the present invention minimizes the influence of environmental stimulus such as pollution, sunshine, free radical and pressure.

An embodiment of the invention provide prevention and/or improve the compositions and the application process thereof of dermatosis and influence thereof.

An embodiment of the invention provide such compositions and application process thereof: the influence that its prevention and reduction produce reactive oxygen species.For example, the present invention includes the application of the activating agent that derives from plant, so that small part is isolated or inhibition arNOX activity.Further, the present invention considers other application synthetic and native compound, with isolated arNOX activity.

The present invention discloses such compositions: its treatment skin and the visible sign and air slaking skin such as wrinkle, microgroove, sagging, pigmentation and the senile plaque that delay actual aging.The present invention also reduces the apparent and situation of sensitivity, drying and/or thin skin skin, with slow redness and/or the sensitive skin of relaxing, and treatment speckle, pimple, scar and other skin blemishes.

The invention provides medicine or cosmetic composition, application process and medicine or cosmetic agents box, be used for changing the disease that causes by the relevant isotype treatment of aging of targeting nadh oxidase (arNOX) by the cellular oxidation that causes aging, described isotype flows into serum by senile cell.Compositions can comprise the reagent from plant extract.For example, compositions of the present invention can comprise at least a extract, described extract shows and suppresses the arNOX activity, no matter separately or with other inhibitor, and suppresses or hinders the activity that flows into the old and feeble relevant isotype of nadh oxidase of serum by senile cell to small part.Compositions can comprise: ubiquinone, natural extract or reagent therefrom, and this natural extract is known to be comprised suppressing the useful activating agent of arNOX; Prepare unguentum, washing liquid, lubricant, gel and analog with other chemical compounds known in the art.These other chemical compounds can comprise natural gum, filler, antiseptic and analog.

In the embodiment, can be with these component portions or all combine with other compositions that often are found in defying age and the repair serum preparation.Medium outside being different from water or dewatering, can comprise liquid or solid lubricant, solvent, wetting agent, thickening agent and powder.Medium can be 0.1% to 99.9% of a composition weight, and is preferred 25% to 80%, and can form composition equilibrated under other improve looks the additive disappearance.In one embodiment, by medium-weight, medium at least 80% is a water.In another embodiment, by weight between about 50% to 85% of water formation compositions.Also have in another embodiment, water exists with about 0.1% to 55% of composition weight.In other embodiments, use other media with above-mentioned concentration.

Depend on the average hydrophil lipophil balance (HLB) of used emulsifying agent to a great extent, oil or oily mater can coexist with emulsifying agent, so that Water-In-Oil or O/w emulsion to be provided.

Compositions of the present invention also can comprise sunscreen.Sunscreen comprises those materials that is usually used in shielding of ultraviolet.Exemplary compounds is derivant, cinnamate and the Salicylate of PABA.For example, octyl methoxycinnamate and 2-hydroxyl-4-methoxy benzophenone (being also referred to as oxybenzone) is available.Octyl methoxycinnamate and 2-hydroxyl-4-methoxy benzophenone obtain by the commercial channel with trade (brand) name Parsol MCX and Benzophenone-3 respectively.The accurate amount of the sunscreen of using in the emulsion can change according to required sun UV radiation protection degree.

Lubricant further can be introduced cosmetic composition of the present invention.The level of this lubricant can be in 0.5% to 50% scope of total composition weight, between preferred 5% to 30%.Lubricant can be categorized as ester, fatty acid and alcohol, polyhydric alcohol and hydrocarbon with general chemical species.

Ester can be monoesters or diester.Acceptable fatty acid diester example comprises dibutyl adipate, ethyl sebacate, diisopropyl two polyester (diisopropyl dimerate) and dioctyl succinate.Acceptable side chain fatty acid ester comprises 2-ethyl-hexyl-myristate, isopropyl stearate and Palmic acid isostearate.Acceptable tribasic ester comprises three linoleic acids, three isopropyl esters and citric acid three lauryls.The acceptable branched chain fatty acid ester comprises Palmic acid lauryl, lactic acid Fructus Amomi Rotundus ester and oleic acid stearate.Preferred ester comprises cocos nucifera oil caprylate/decanoin (mixture of cocos nucifera oil caprylate and cocos nucifera oil decanoin), propylene glycol Fructus Amomi Rotundus ether acetate (propylene glycol myristyl ether acetate), diisopropyl adipate and Octanoic acid, hexadecyl ester.

Suitable aliphatic alcohol and acid comprise those chemical compounds with 10 to 20 carbon atoms.Particularly preferably be the chemical compound as the pure and mild acid of cetyl, myristyl, palmityl and stearyl.

In these polyhydric alcohol, what it can be used as lubricant is straight chain and branched chain alkyl polyhydroxyl compounds.For example, preferred propylene glycol, sorbitol and glycerol.Polymerized polyalcohol is also available, as polypropylene glycol and Polyethylene Glycol.Butanediol and propylene glycol are also especially preferably made penetration enhancer.

The exemplary hydrocarbon that can be used as lubricant is the chemical compound with any 12 to 30 carbon atom hydrocarbon chains.Instantiation comprises mineral oil, vaseline, Squalene and isoparaffin.

Other embodiments of the present composition comprise thickening agent.Thickening agent will be usually with 0.1% to 20% of composition weight, and preferably about 0.5% to 10% any amount with composition weight exists.Exemplary thickening agent is crosslinked polyacrylate material, with trade (brand) name CARBOPOL

Available from B.F.Goodrich Co..Can use natural gum, as xanthan gum, carrageenan, gel, karaya, pectin and locust bean gum.Under certain conditions, can realize thickening function by the material that also is used as silica gel or lubricant.For example, surpass the silicone adhesive of 10 centistokes and have difunctional such as the esters of tristerin.

Powder can be introduced cosmetic composition of the present invention.

These powder comprise the Magnesiumaluminumsilicate of Chalk, Talcum, Kaolin, starch, smectite clay, chemical modification, organically-modified montmorillonitic clay, aluminium hydrosilicate, pyrogenic silica, starch ocentyl succinic aluminum and composition thereof.

Also other additional accessory constituents can be mixed cosmetic composition.These compositions can comprise coloring agent, opacifier and flavouring agent.The amount of these other additional accessory constituent can be at random in 0.001% to 20% scope of composition weight.

Compositions of the present invention can be used for the topical application of human skin, as regulating, preserve moisture and level and smooth skin, increases the reagent of the appearance of elasticity (flexibility and elasticity) and prevention or minimizing wrinkle, microgroove or old and feeble skin.Preparation of the present invention provides the response to the skin color loss, and facilitate the hydration of effective promotion surface skin layer and firm, meanwhile be devoted to repair the skin bottom to help to reduce microgroove and wrinkle and occur and to recover visible hue and elasticity with antioxidant and other beneficiating ingredients.In some illustrative embodiments, these antioxidants are especially at suppressing arNOX.

The compositions that can comprise in one embodiment, about 1 to 1000ml activating agent on a small quantity to dermal administration.In the exemplary embodiment, can be to a certain amount of compositions that comprises about 1 to 100ml activating agent of dermal administration.This process can repeat to continue any a period of time several times in every day.Preferably, to skin early once with late applied once compositions.

Local skin care composition of the present invention can be mixed with washing liquid, unguentum, gel or analog.The proper container of compositions can being packed into is to mate with the intended use of its viscosity and user.For example, washing liquid or unguentum can be packed into bottle or roller ball applicator, or the aerosol device of propellant actuated or the container of the pump that is suitable for finger manipulation is housed.When compositions was unguentum, it can simply be stored in non deformable bottle or the squeeze receptacle, as managing or have the jar of lid.Therefore the present invention also provides the beauty treatment that contains this paper qualification can accept the closed container of compositions.

In the mode of explanation, be not to provide following examples with ways to restrain.

Embodiment

Embodiment 1

The sign of narcissus extract

In order to check narcissus may have the hypothesis of arNOX inhibitor, (Brent and Becky ' s Bulbs, Gloucester VA) obtain the bulb of pure white narcissus (Narcissus tazetta) from provider.Preparation suppresses arNOX water and alcohol extract active and that enhancing suppresses when in conjunction with low activity narcissus powder (Xian Aojing Science and Technology Developing CO., LTD, Xian, China).(potentiation MO) is shown in Fig. 1 a for Sigma-Aldrich, St.Louis to derive from the powder of narcissus bulb and SHAM.By following identical scheme, determine that naractins strengthens the ability that arNOX suppresses, but use salicin (Sigma-Aldrich) and derive from narcissus powder (the Xian Aojing Science and Technology Developing CO. of flower, LTD, Xian, China) suppress Fig. 1 b to strengthen arNOX.With lack the similar extract that arNOX suppresses active Narcissus pseudo-narcissus (daffadowndilly) and Narcissus jonquilla (daffodil) extract and compare.As shown in Figure 2, the activity of narcissus extract is with relevant by adding ferric iron enhanced red colored---characteristic of naturally occurring hydroxamate---, this hydroxamate be found in corn (corn (maize)) (Fig. 3) and Herba bromi japonici (not shown) rice shoot in, suppress arNOX (Fig. 4) equally.By thin-layer chromatographic analysis (Fig. 5), reach spectral investigation from its ferrum adduct, and also from Zea mays root (Fig. 6) dimension body (the central vasculature of Zea mays root, wherein hydroxamate be concentrated and corresponding to the potential rich hydroxamate stem vasculature of narcissus bulb shown in Figure 2) and by with commercially available hydroxamate (salicylhydroxamic acid salt (SHAM), Sigma-Aldrich) contrast provides the representative of narcissus extract active component the natural further evidence that has the potentiality of hydroxamate.

Detect the arNOX activity inhibition of SHAM---the optional oxidase active inhibitor of known plant---, under concentration 1mg/ml, obtain being higher than active suppress (Fig. 7) of arNOX of 72 years old male's saliva of 90%.Find that SHAM also strengthens the active inhibition of the active narcissus powder of part arNOX (Fig. 1).Commercially available narcissus powder (IBR-DORMIN

, Israel) present really and contain low-level naturally occurring hydroxamate, this hydroxamate may be similar to SHAM but on far below the level of finding (Fig. 5 a and 5b) in by the made extract of bright narcissus bulb.

Embodiment 2

Determining of arNOX inhibitor

Whether the arNOX activity is suppressed important problem in order to answer the salicylic acid part of stablizing the salicyl hydroxamate, salicylic acid (Naractin 2) is detected as 72 years old active inhibitor of male's saliva arNOX, and find that it suppresses arNOX activity (Fig. 8).When reacting with ferric iron, similar to what hydroxamate presented, salicylic acid also takes on a red color.Salicylate and/or aspirin do not suppress the arNOX activity.Detect the water extract of bark of willow---natural origin of Salicylate---, and find that it also suppresses arNOX.Also detected glucosides salicin (Naractin 3)---the main Salicylate of bark of willow subsequently, and found that it has activity (Fig. 9) under the concentration than low 10 times of SHAM (Naractin 1) or salicylic acid (Naractin 2).

Embodiment 3

Produce the evaluation of the plant of Salicylate

Study to identify plant as the Salicylate natural producer.Be applied in the Natural Products data base that University of Illinois in Chicago creates by Norman professor Farnsworth (napralert.org can with) study.

Embodiment 4

The potentiation of arNOX inhibitor

Salicin not only has activity (Fig. 9) as single agents, and it also has such activity: strengthen from the arNOX inhibitory action (Figure 10 a and 10b) such as other arNOX inhibitor mixed things of the natural origin of Chinese Magnolivine Fruit and narcissus powder.Shown in Figure 10 a, show that the 4mg/ml Chinese Magnolivine Fruit adds the arNOX inhibitory action of the mixture of narcissus extract (20 μ l) and additional 1mg/ml salicin.Fig. 1 b is illustrated in the 1mg/ml salicin and exists down, and the 4mg/ml Chinese Magnolivine Fruit adds the arNOX inhibitory action of the mixture of 1mg/ml narcissus powder.(be used to represent wherein any one term of several natural arNOX of existence inhibitor through three kinds of chemical pures " Naractins " of identifying and detect, this the natural arNOX of existence inhibitor is present in the narcissus powder maybe can strengthen the inhibition level of narcissus powder to the active bright narcissus extract of tool that is equal to) in, salicin presents most promising.Salicin is stable, water soluble, nonirritant, relatively cheap and can be from a plurality of commodity provider, as Sigma-Aldrich, and St.Louis, Mo. obtains.Further, the clear enhancing that shows low activity narcissus powder of Fig. 1 a and 1b, wherein SHAM and salicin all significantly reduce the arNOX activity.

Embodiment 5

The sign of arNOX

The peroxide that is caused by dark yellow cover layer (Buffy coat) produces: dark yellow cover layer, lymphocyte and hematoblastic mixture.This dark yellow cover layer commercial can available from, for example Rockland ImmunoChemicals (Gilbertsville, PA).Before the Collection and analysis blood sample is remained on 4 ℃.With Ca.10 ⁷Cell adds in each test.Use hematimeter and measure cell quantity.

Measurement standard (the Mayo that the minimizing of the ferricytochrome c that application is caused by peroxide forms as peroxide, L.A and Cumutte, J. (1990) Meth.Enzyme.186,567-575.7.Butler, J, Koppenol, W.H. and Margollash, E. (1982) J.Biol.Chem.257,10747).When suppressing coupling with the generation of measurement peroxide with superoxide dismutase, this is extensive received method.This test is by PBSG buffer (8.06g NaCl, 0.2g KCl, 0.18g Na ₂HPO ₄, 0.26g KH ₂PO ₄, 0.13g CaCl ₂, 0.1g MgCl ₂, the 1.35g glucose is dissolved in the 1000ml deionized water, is adjusted to pH7.4, filter and in 4 ℃ of preservations) in 150 μ l serum or the dark yellow cover layer of 40 μ l form.(NY USA) measures speed under the dual wavelength operation pattern for Milton Roy, Rochester, and continuous detecting was 1 minute in per 1.5 minutes to use SLM Aminco DW-2000 spectrophotometer.After 45 minutes, add detection compound, and reaction continues 45 minutes.After 45 minutes, use mM extinction coefficient 19.1cm ^-1Detect the ferricytochrome c that reduces.Table 1 provides the result of detection compound.Unless shown in having in addition, extract is made by this chemical compound.

Table 1 provides some arNOX to suppress result of experiment.

Table 1

Embodiment 6

The local beauty goods

With regard to its effectiveness and toleration with respect to two kinds of (2) vehicle Control, carry out the controlled clinical application research of eight all broken faces to screen four kinds of (4) prototype anti-senile preparations, said preparation comprises the plant extract with arNOX rejection characteristic.Measure (chromascope (Chroma Meter), epidermis water content detector (Corneometer), skin elasticity detector (Cutometer)) and self assessment investigation (Self-Assessment Questionnaires) evaluation effectiveness by clinical scale, biological plant.Estimate toleration by stimulating classification and monitoring adverse events.

Totally 23 subjectss finish and participate in research.The zone has and slight have the qualified participation of Pigmented subjects to medium microgroove and coarse groove and facial right side and left side and study near the eyes.According to design at random, the subjects is assigned to two kinds of following detection material (a kind of is contrast, and a kind of is detection material):

Contrast:

A.arNOX control gel A (no label), AB-87-04A, colourless, clear gel (12 subjectss)

B.arNOX control gel B (red label), JZ-91-40, colourless, clear gel (containing glycerol) (11 subjectss)

Detection material:

1.arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green label), JZ 91-39, pink, clear gel (6 subjectss)

2.arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (blue label), TL-90-59, colourless, clear gel (6 subjectss)

3.arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (yellow label), TL-90-58 (containing glycerol), pink, clear gel (6 subjectss)

4.arNOX control gel B w/ salicin (half yellow label, half black label), KK-89-49, colourless, clear gel (containing glycerol) (5 subjectss)

Use the assigned detection material to facial right side or left side twice of indication subjects every day (early and evening) behind its face of cleaning, and use distribution to shining facial offside.

Carry out clinical evaluation in baseline (observing 1), the 4th week (observing 2) and the 8th week (observing 3).The subjects participates in following clinical scale and device program (unless shown in having in addition) when each the observation.

Effectiveness/performance parameter

With regard to following parameters the subjects is carried out clinical scale on facial right side and left side: microgroove (near the eyes), coarse groove (near the eyes), skin texture (buccal), whole variable color, brightness (buccal), skin transparency, pore size (forehead and nasal area), pore distribution/structure and whole skin radiation (gloss).

The classification of stimulation/security parameter

With regard to objective stimulus parameter (erythema, edema, desquamation) and subjective irritation parameter (calcination, acupuncture, pruritus, feel oppressed, fiber crops thorn) subjects is carried out clinical scale on facial right side and left side.

The skin surface hydration is measured

Use epidermis water content detector Corneometer

CM 825 (Courage+Khazaka, Germany) bearing analysis device carries out skin surface and contains water gaging.Measure (three times) at left cheek and the low center of right cheek, with quantitative horny layer moisture content.

The skin brightness measurement

Use chromascope Chroma Meter CR400 (Konica-Minolta, Japan) skin brightness analysis device carries out the skin brightness measurement three times, and carries out the skin brightness measurement with the variation by the appraisal of equipment skin color/tone in the pigment sexually transmitted disease (STD) change (being selected by researcher) in facial right side and left side.Carry out other skin brightness measurement in facial side non-pigment (normally) zone.

The skin viscoelasticity measurement

Application skins based upon bidding elastomeric check instrument Cutometer

SEM 575 (Courage+Khazaka, Germany) viscoelastometer carries out independent skin viscoelasticity measurement.Center at right cheek of each subjects and left cheek is measured, with the viscoelasticity of evaluating skin.

Investigation

The subjects is in the 4th week and the 8th week finished following investigation.

Subjects's skin that research beginning back changes about the skin parameter changes estimates (Subject Skin Change Evaluation) investigation

Estimate investigation (Subject Evaluation Questionnaire) about the present case of skin parameter and the subjects of detection material attribute and toleration

Generally, this result of study shows when comparing with the baseline mark, all detection material and to impinging upon microgroove is apparent, all produce significantly on sense of touch roughness, skin color and the whole variable color/pigmentation and improve; It is apparent that detection material 1---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green label) JZ-91-39 and detection material 3---arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (yellow label) TL-90-58 (containing glycerol) improve coarse groove.Detection material or contrast any objective or subjective irritation and obviously do not increase.

The skin brightness measurement shows only detection material 1, the b of arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green) JZ-91-39 ^*Value produces obviously minimizing in the non-pigment position in the 8th week.Put the skin brightness b that is got in the displacement of pigment sexually transmitted disease (STD) ^*---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation), narcissus extract and salicin (green) JZ-91-39 are reducing pathological changes---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (indigo plant) TL-90-59 and detection material 3---the arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation) that is better than detection material 2 on obscure, narcissus extract and salicin (Huang) TL90-58 (containing glycerol) to value representation detection material 1.The skin surface hydration measure to show, all detection material and significantly improve skin hydration to impinging upon for the 4th week.Viscoelasticity measurement does not show any significant difference between detection material and the contrast.

Informed Consent Form

Before the research registration, obtain to meet the written Informed Consent Form of 21 CFR 50.25 (coding of Federal Regulations) from each subjects.The Informed Consent Form original paper that participates in each subjects's signature of this research can be retained in the research file.Each subjects receives signed letter of consent duplicate.(seeing also appendix IV sample table)

Loss

23 (23) individual subjectss finish this research.26 (26) individual subjectss register and participate in this research, and three (3) subjectss participate in this research because underlying cause withdraws from:

Withdraw from voluntarily/adverse events: the subjects 021

Can not participate in predetermined investigation: subjects 004 and 026

Withdraw from voluntarily/the stroke conflict: 020,022,029

Can not participate in predetermined investigation: 009,034

Researcher decision: 010

Subjects's demography

23 (23) individual women subjectss finish this research.Table 2 provides all subjects demographic informations the summary of (age, race and the classification of Fitzpatrick skin).For race and Fitzpatrick skin type, listed all kinds of subjectss' quantity, in the bracket percentage rate of subjects's number.Ethnic information obtains from each subjects's qualification and health survey (Eligibility and Health Questionnaire).

Table 2: the demographic information sums up

The classification of Fitzpatrick skin does not have the unprotect reaction that the sun is exposed to the sun and afterwards the preceding 30-45 minute sun is exposed to the sun in the winter time based on skin.The kind of skin type is as follows:

Type i. easily calcination all the time; Never tanned (tan);

Type II. easily calcination all the time; Minimum level is tanned;

Type-iii. medium calcination; Tanned gradually;

The calcination of type i V. minimum level; All the time fully tanned

Type V. is calcination hardly; Tanned fully;

Type VI. is never tanned; The degree of depth forms the color element;

Fitzpatrick has reported optional categorizing system, and it is used for estimating the degree of mouthful week and eye socket (near the eyes) wrinkle (rhytidosis):

I level-microgroove;

II level-microgroove is to the stricture of vagina line of medium dark stricture of vagina and moderate quatity;

III level-microgroove is to dark (slightly) stricture of vagina, in a large number streaklines and may have many gauffers

Embodiment 7

Program and method

Before the research beginning, the expection subjects participates in three days eluting phases, during do not use facial wetting agent to face.

Baseline (observe 1), the subjects of expection cleaned its face at least arriving at the clinic before 30 minutes, and removed all cosmetic product.The subjects of expection brings its conventional skin nursing method product to consider to carry out qualification.The subjects finishes qualification and health survey, and the signature Informed Consent Form, confidentiality agreement (Confidentiality Agreement) and photography issue form (Photography Release Form).

The subjects participates in following clinical scale program:

Effectiveness/performance parameter

With regard to following parameters, the subjects is carried out clinical scale on facial right side and left side:

Microgroove-regional near the eyes

Coarse groove-regional near the eyes

Skin texture (as seen apparent)-buccal

Sense of touch roughness-buccal

Whole variable color

Whole skin gloss

Use the fractionated result of following 1 to 10 graduation record effectiveness/performance parameter:

1=is (1 to 3=good/expectation) just

10=bears (8 to 10=non-expectations)

As needs, use the half point mark

The subjects slight continues research to the qualified participation of middle isodisperse by having, and this mark is microgroove 3 to 7 near the eyes; Coarse groove 2 to 5 near the eyes; With pigmentation on facial right side and the left side 2 to 7.

The classification of stimulation/security parameter

With regard to objective stimulus parameter (erythema, edema, desquamation) and subjective irritation parameter (calcination, acupuncture, pruritus, feel oppressed, fiber crops thorn) subjects is carried out clinical scale on facial right side and left side.Use following grade record and stimulate fractionated result:

0=does not have

1=is slight

2=is medium

3=is serious

If necessary, use half point

Titular subjects participates in following device measuring:

Embodiment 8

The skin surface hydration is measured

Use epidermis water content detector Corneometer

CM 825 (Courage+Khazaka, Germany) bearing analysis device carries out skin surface and contains water gaging.Measurement is carried out three times and is carried out at the low center of left cheek and right cheek, with quantitative horny layer moisture content.Epidermis water content detector Corneometer

Measuring principle be based on dielectric capacitance measurement.Any change in dielectric constant that is caused by the moisture variation of skin surface changes the electric capacity of accurate measurement capacitor.These measurements can detect the most small variation of skin aqueous level, and high repeatability is arranged.Reading is directly proportional with the skin capacitance, and measured value increases with skin is more moistening.

Embodiment 9

The skin brightness measurement

Application chromascope Chroma Meter CR400 (Konica-Minolta, Japan) skin brightness analysis device carries out the skin brightness measurement three times and carry out the skin brightness measurement in the pigment sexually transmitted disease (STD) change (being selected by researcher) in facial right side and left side.The variation of chromascope equipment ground (and objectively) evaluating skin color/tone.Carrying out other chromascope in facial side non-pigment (normally) zone measures.Chromascope is sensitive colorimeter, the setting of its acceptability of color matches color of object and calibration.Chromascope has separable head, to be easy to and independent analysis institute favored area.The following value of record:

L ^*: to describe relative lightness from black extremely white gray scale; Value becomes bright more and bright more with skin to be increased

a ^*: to green scope, the color accent is described from red; The improvement that value generates with skin heart, blood flow increase and the skin color raising increases

b ^*: describe color in from indigo plant to yellow scope and transfer; Value generally becomes bright with skin and reduces.

Carry out additional skin brightness measurement in facial side non-pigment (normally) zone of each subjects.

Embodiment 10

The skin viscoelasticity measurement

Application skins based upon bidding elastomeric check instrument Cutometer

SEM 575 (Courage+Khazaka, Germany) viscoelastometer carries out independent viscoelasticity measurement.Center at right cheek of each subjects and left cheek is measured, with the viscoelasticity of evaluating skin.Measuring principle is based on suction.Create negative pressure in the device, skin is inhaled in the probe aperture.In probe, measure penetration depth by untouchable optical measuring system.Light intensity is owing to the penetration depth of skin changes.At the resistance (rockiness returns the ability (elasticity) of its original position with it) that when each measures end, sucks skin on the equipment with the curve display negative pressure.Use the 300mbar negative pressure, and discharge by the probe of 8 millimeters (mm).In the application of suction and the motion of deenergized period record skin turnover probe, and calculating elastic and extensibility.

Distribute to the wherein a kind of of the following detection material group of subjects according to designing at random:

Contrast

A.arNOX control gel A (no label)---AB-87-04A

B.arNOX control gel B (red label) JZ-91-40 (containing glycerol)

Detection material

1.arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green label) JZ91-39

2.arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (blue label) TL-90-59

3.arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (yellow label) TL-90-58 (containing glycerol)

4.arNOX control gel B w/ salicin (half yellow label, half black label)---KK-89-49 (containing glycerol)

The subjects uses the assigned detection material to facial right side or left side according to the indication of following operation instruction, and use distribution to shining facial offside:

Twice of every day used thin layer behind the face that sooner or later cleans you.

After using, detection material can use wetting agent and cosmetics.

The written operation instruction of subjects, the following diary of observing calendar and record detection material application times and note are provided.

The subjects returns the clinic in the 4th week (observation 2) and the 8th week (observing 3), cleans its face at least before 30 minutes and removes all cosmetic product in each observation.The clinic is returned in diary and detection material, and check and use adaptability; At the new diary book of the 4th week distribution (and detection material, as needs).According to the baseline program, the subjects accepts clinical scale and biological plant is measured (chromascope, epidermis water content detector and skin elasticity detector).Subjects's skin variation evaluation investigation and subjects that each subjects also will finish about detection material attribute, toleration and facial right side and the improvement of left side skin estimate investigation.

Table 3 provides the batching of each compositions, and is as follows:

^**Noveon?IP?Holdings?Corp.Cleveland，Ohio，U.S.

Clariant，Corp.Charlotte，N.C.，U.S.

Angus?Chemical?Co.，Buffalo?Grove?Il，U.S.

Unigema，New?Castle，DE，U.S.

Symrise?Inc.，Teterboro，NJ

Draco?Natural?Products，Inc.，San?Jose，CA，U.S.A.

Xuancheng?Baicao?Plants?Industry?and?Trade?CO.，LTD，Anhui，China

Offer the written operation instruction of subjects, the following diary of observing calendar and record detection material number of applications and note.

The subjects returns the clinic in the 4th week (observing 2) and the 8th week (observing 3).The subjects cleaned before going to testing agency to observe that it is facial and remove cosmetic product at least in 30 minutes at every turn.The subjects also will take their detection material to each observation, uses adaptability to check.The subjects participates in follow procedure when each the observation:

The classification of effectiveness/performance parameter

The classification of stimulation/security parameter

Moisture (the epidermis water content detector Corneometer of skin surface ) measure

Skin brightness (chromascope Chroma Meter) is measured

Skin viscoelasticity (skin elasticity detector Curtometer ) measure

Subjects's skin variation evaluation investigation and subjects that the subjects also will finish about detection material attribute, toleration and facial right side and the raising of left side skin parameter estimate investigation.

During each the observation clinic is returned in diary, and distributed new diary book at 2 o'clock in observation.The subjects returns the detection material unit to the clinic when finishing this research.The clinic personnel examine diary, and weigh the detection material unit to guarantee adaptability when each the observation.

Embodiment 11

Biometrics and data management

With the pairing t-under p≤0.05 significance level check (t-test) with the 4th week (observing 2) and the 8th week (observing 3) clinical scale and the meansigma methods and baseline (observation 1) meansigma methods of device measuring contrast statistically.For all properties, calculating changes and improves incidence rate from the mean percentage of baseline.Using variance analysis (ANOVA) and paired comparison (Fisher ' s LSD) compares between detection material and contrast.

The subjects is tabulated in the 4th self evaluation investigation all and that finished in the 8th week, and carry out top box (top box) analysis.

Embodiment 12

The local application data

In baseline (observing 1), the 4th week (observing 2) and the 8th week (observing 3), the subjects carries out clinical scale and biological plant measurement (chromascope, epidermis water content detector and skin elasticity detector) at face.Table 4 shows the result of each detection material and contrast.The meansigma methods and the baseline average in the 4th week and the 8th week are carried out the statistics contrast, obtain significant difference.List in the bracket from the mean percentage of baseline and change.

Finger is compared significantly (p≤0.05) minimizing of statistics with baseline

Finger is compared significantly (p 〉=0.05) increase of statistics with baseline

^*Owing to rearrange Equipment Alignment mistake when observing, subjects 010 is removed from the 8th all skin elastomeric check instrument are analyzed; The 8th all skin elastomeric check instrument are measured the n value and are equaled 11.

Embodiment 13

Mean change from baseline

Table 5 provides the comparison from the baseline mean change of clinical scale and equipment research.

Table 5

Table 5 (continuing)

Embodiment 14

Compare between group

Based on mean change, use variance analysis (ANOVA) and paired comparison (Fisher ' s LSD) and between treatment (detection material and contrast), compare from baseline.The following classification declaration that provides in the table 6 remarkable (p≤0.05) difference of statistics between experimental group.To minimum order performance classification situation of improving the standard, the parameter of no significant difference is not listed with maximum.Under each treatment, list mean change from baseline.

Table 6

Embodiment 15

The statistics comparative result of Equipment Inspection

The subjects is carried out classification with microgroove (near the eyes), coarse groove/skinfold (near the eyes), sense of touch roughness (buccal), skin color, whole variable color/pigmentation and whole skin gloss on facial right side and the left side.When comparing with baseline, this clinical scale result shows following remarkable improvement, and is as shown in table 7.

Refer to significantly (p≤0.05) minimizing of statistics, the improvement of comparing with baseline

The 4th week of W4=

The 8th week of W8=

Embodiment 16

Stimulation/safety

The subjects by with the erythema on facial right side and the left side, edema, desquamation, calcination, acupuncture, pruritus, feel oppressed and fiber crops thorns carries out classification.

When comparing with baseline, the clinical scale result shows significant improvement, and is as shown in table 8, for example:

Point out that the statistics of comparing with baseline remarkable (p≤0.05) reduces---improve

The 4th week of W4=

The 8th week of W8=

Embodiment 17

Skin brightness detects

Application skins based upon bidding brightness analysis device (chromascope Chroma Meter CR400, Konica-Minolta, Japan) is measured skin brightness three times.Pigment sexually transmitted disease (STD) to facial right side and left side becomes (being selected by researcher) to be measured, with the variation through the appraisal of equipment skin color/tone.Carry out other skin brightness measurement in facial side non-pigment (normally) zone.

Table 9 shows the result of skin brightness measurement, and the significant difference of demonstration when comparing with baseline, for example:

The statistics that finger is compared with baseline significantly (p≤0.05) reduces

The statistics that finger is compared with baseline significantly (p≤0.05) increases

The 4th week of W4=

The 8th week of W8=

Embodiment 18

The moisture detection of skin surface

Carry out skin surface and contain water gaging three times (epidermis water content detector Corneometer CM 825, Courage+Khazaka, Germany).Measure at right cheek and the low center of left cheek, with quantitative horny layer moisture content.

Skin surface contains water gaging and shows when comparing with baseline, respectively treats the remarkable increase (improvement) of moisture-keeping function the 4th week.The 8th week was not found significant difference.

Embodiment 19

The skin viscoelasticity measurement

Application skins based upon bidding elastomeric check instrument Cutometer MPA 580 (Courage+Khazaka, Germany) carry out independent skin viscoelasticity measurement.Center at right cheek of each subjects and left cheek is measured, with the viscoelasticity of evaluating skin.

Following significant difference when the viscoelasticity measurement result demonstration shown in the table 10 is compared with baseline, for example:

The 4th week of W4=

The 8th week of W8=

Embodiment 20

Overall conclusion

The result of this pilot study shows when all detection material are compared with the baseline mark with contrast, produces significantly on microgroove, sense of touch roughness, skin color and whole variable color/pigmentation are apparent and improves.Pay special attention to, detection material 1---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation), narcissus extract and salicin (green label) JZ-91-39 and detection material 3---arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation), narcissus extract and salicin (yellow label) TL-90-58 (containing glycerol) even have a good effect to coarse groove is apparent, in the 4th week (detection material 3, yellow label) and the 8th week (detection material 1, green label; With detection material 3, yellow label) is remarkable improvement.Any detection material or to impinge upon objective or subjective irritation on significantly do not increase.

The skin brightness measurement shows only detection material 1---the b of arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green label) JZ-91-39 ^*Value is positioned to produce obviously in the 8th week in non-pigment portion and reduces.Become the skin brightness b that get at the position in the pigment sexually transmitted disease (STD) ^*Value shows detection material 1, and---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation), narcissus extract and salicin (green label) JZ-91-39 are reducing pathological changes---arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (blue label) TL-90-59 and detection material 3---the arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (encapsulation) that is better than detection material 2 on obscure, narcissus extract and salicin (yellow label) TL-90-58 (containing glycerol).

The moisture measurement of skin surface shows, all detection material and significantly improve the skin hydration effect to impinging upon for the 4th week.

The skin viscoelasticity measurement does not show any significant difference between detection material and the contrast.

ANOVA between detection material and the contrast comparison shows that detection material 1 (arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (green label) JZ-91-39) and detection material 2 (arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (blue label) TL-90-59) are better than contrasting A (arNOX control gel A (no label) AB-87-04A) and contrast B (arNOX control gel B (red label) JZ-91-40 (containing glycerol)) in improving in the 4th week in the whole variable color.Further, detection material 1 and 2 is in being better than contrasting B (arNOX control gel B (red label) JZ-91-40 (containing glycerol)) the 8th week.Detection material 2 (arNOX control gel A w/ Fructus Schisandrae Chinensis (Schizanda) (blue label) TL-90-59), detection material 3 (arNOX control gel B w/ Fructus Schisandrae Chinensis (Schizanda) (not encapsulation), narcissus extract and salicin (yellow label) TL-90-58 (containing glycerol)) and detection material 4 (arNOX control gel B w/ salicin (half yellow label, half black label) KK-89-49 (containing glycerol)) on improvement is apparent, is better than contrasting A (arNOX control gel A (no label) AB-87-04A) and contrast B (arNOX control gel B (red label) JZ-91-40 (containing glycerol)) in the 8th week.

Claims

1. topical composition that is used to improve aging effect comprises:

At least a arNOX inhibitor of effective dose,

Wherein said arNOX inhibitor is naractin, and wherein said naractin effectively reduces described aging effect.

2. the described compositions of claim 1, wherein said naractin derives from plant extract.

3. the described compositions of claim 2, wherein said naractin is the Salicylate or derivatives thereof.

4. the described compositions of claim 2, wherein said naractin purifies from narcissus (N.tazetta), willow, corn, crepis, willow, Caulis Viburni Dilatati, aspergillosis (Aspergillus), Radix Alangii, birch, Radix Bupleuri, Colchicum autumnale, the Radix Euphorbiae Pekinensis, filipendulum, Fructus Gardeniae, Radix Arnebiae (Radix Lithospermi), Nicotiana tabacum L. or Herba Visci.

5. the described compositions of claim 3, wherein said Salicylate is salicin, salicylic acid, salicylhydroxamic acid salt or its combination.

6. the described topical composition of claim 1, wherein said compositions further comprise beauty treatment or pharmaceutically acceptable carrier.

7. the described topical composition of claim 1 wherein exists more than a kind of described arNOX inhibitor, and described more than a kind of described arNOX inhibitor form that is plant extract.

8. the described topical composition of claim 7, wherein said plant be selected from Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae (Oenothera biennis), astaxanthin, tangerine, Fructus Schisandrae Chinensis (Schisandra chinesis), Radix Ophiopogonis (Lonicera), Semen Fagopyri Esculenti (Fagopyrum), Radix Dauci Sativae, narcissus (Narcissus tazetta), Fructus Canarii albi, willow, Herba bromi japonici or corn.

9. the described topical composition of claim 8, wherein said Radix Ophiopogonis is Flos Lonicerae (Lonicera japonica) or the Flos Lonicerae tragophyllae of overgrowing (Lonicera caprifolium).

10. the described topical composition of claim 7, wherein said arNOX inhibitor is beta-carotene or astaxanthin.

11. the described topical composition of claim 7, wherein said naractin increases the effect of additional arNOX inhibitor.

12. the described topical composition of claim 1 is wherein with unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen give described compositions.

13. the described topical composition of claim 1, wherein said aging effect comprises: microgroove, wrinkle, pigmentation, dewater, follow the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

14. the described topical composition of claim 1 wherein provides described arNOX inhibitor with the concentration of about 5 μ g/ml between about 500 μ g/ml.

15. an inhibition is produced to improve the method for aging effect by the reactive oxygen species that the old and feeble relevant isotype of nadh oxidase causes, comprising:

Comprise salicin, salicylic acid, wherein at least a compositions of salicylhydroxamic acid salt to what its required patient treated effective dose, suppress thus to produce, and wherein improve aging effect by the reactive oxygen species that the old and feeble relevant isotype of nadh oxidase causes.

16. the described method of claim 15, wherein said compositions further comprise from Brassica oleracea L. var. botrytis L., Lentinus Edodes, flame nettle, Herba Rosmarini Officinalis, Flos Nelumbinis, Carlina acaulis, extra large Flos Rosae Rugosae, Citrus, Radix Oenotherae erythrosepalae, astaxanthin, tangerine, Fructus Schisandrae Chinensis, Radix Ophiopogonis, Semen Fagopyri Esculenti, Radix Dauci Sativae, narcissus, Fructus Canarii albi, willow, Herba bromi japonici or wherein at least a extract of corn.

17. the described method of claim 15, wherein said compositions is as unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen are used.

18. the described method of claim 15, wherein said aging effect comprises: microgroove, wrinkle, pigmentation, dewater, follow the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

19. a beauty method that improves aging effect comprises that to the dermal administration cosmetic composition, described cosmetic composition comprises:

Be enough to suppress the effective dose naractin of arNOX,

The aging effect of wherein at least a arNOX mediation is suppressed.

20. the described method of claim 19, wherein said naractin is a Salicylate.

21. the described method of claim 20, wherein said Salicylate are salicin, salicylhydroxamic acid salt or salicylic acid.

22. the described method of claim 19, wherein said cosmetic composition further comprises plant extract, comprising: Radix Dauci Sativae extract, Fructus Canrii Albi extract, cauliflower extract, Lentinus Edodes extract, flame nettle extract, Herba Rosmarini Officinalis extract, Flos Nelumbinis extract, Carlina acaulis extract, extra large rose extract, Citrus reticulata Blanco extract, Radix Oenotherae erythrosepalae extract, tangerine extract, Fructus Schisandrae Chinensis extrat, Radix Ophiopogonis extract, Semen Fagopyri Esculenti extract, willow extract, corn dimension body, Herba bromi japonici dimension body or narcissus extract.

23. the described beauty method of claim 19, wherein said naractin provides with the beauty treatment acceptable carrier.

24. the described beauty method of claim 19, wherein said aging effect comprises: microgroove, wrinkle, pigmentation, dewater, follow the string, hemangioma, drying, pruritus, telangiectasis, actinic purpura, seborrheic keratosis, shortage aquation, collagen protein reduce or actinic keratosis.

25. the described beauty method of claim 19, the wherein at least one day described naractin of applied once.

26. the described beauty method of claim 19 wherein provides described naractin with the concentration of about 5 μ g/ml between about 500 μ g/ml in beautification product.

27. the described beauty method of claim 19 is wherein with unguentum, emulsion, washing liquid, gel, the suspension of lipid or polymeric microspheres or nanometer spheroid or vesicle; Soap, shampoo or sunscreen give described compositions.

28. one kind is used to use to improving the test kit of the useful cosmetics of aging effect, comprises:

At least a naractin; With

Operation instruction.

29. the described test kit of claim 28 further comprises the beautification product that is suitable for as at least a arNOX inhibition plant extract carrier.