CN101291679A - Compositions comprising actinidia and methods of use thereof - Google Patents

Abstract

Disclosed are novel preparations of Actinidia, and particularly species of hardy kiwifruit, as well as compositions comprising the same. Also disclosed is the use of these preparations of Actinidia to prevent and/or treat a variety of diseases in which regulation of the immune response is effective, including both allergic and non-allergic inflammatory disease, viral infection, and cancer. Methods related to making and using these compositions are also described.

Description

The compoistion and method of use that comprises actinidia

Invention field

The present invention relates to actinidia (Actinidia), particularly hard kiwi fruit (hardy kiwifruit) species, and various fraction and prepared product, and comprise aforementioned compositions, they all have the ability to prevent and/or treat the effective therein multiple disease of regulation and control immunne response, comprise allergia and nonallergic inflammatory disease, viral infection and cancer.Also describe preparation and used the related method of these compositionss.

Background of invention

The disease that relates to inflammation is characterised in that some cell type and vectorial inflow, and their existence can cause tissue injury, causes death sometimes.In some organ and system, be deleterious especially when suffering from the disease that relates to inflammation such as respiratory system, can cause breathing be obstructed, hypoxemia, hypercapnia and lung tissue damage.In other disease or disease, the generation of some type inflammation may be the pith of disease control, such as in viral infection, although the tissue injury of infected zone is still risky.

Allergic disease is mediated by IgE (IgE) to a certain extent, and auxiliary (Th2) cell of 2 type T, mastocyte and eosinophilic granulocyte have also demonstrated in disease process (the Maggi E. that plays a significant role, Immunotechnology, 3:233-244,1998; Pawankar R., Curr.Opin.Allergy Clin.Immunol., 1:3-6,2001; Vercelli D., Clin.Allergy Immunol., 16:179-196,2002).IgE in the circulation is in conjunction with two kinds of isotypes (isoform) of IgE receptor: the high-affinity IgE receptor that exists on mastocyte and the basophilic leukocyte surface (Fc ε RI), and the low-affinity IgE receptor (Fc ε RII or CD23) that exists on lymphocyte, eosinophilic granulocyte, platelet and the macrophage surface.Think that control allergic conditions pathogenetic key factor is meet with behind the allergen IgE receptor on the mastocyte crosslinked, and the mast cell degranulation that causes thus.The molecule that mastocyte discharges comprises histamine, heparin, protease and free radical, and their mediation various biological effects comprise vasodilation, intestinal and/or bronchial smooth muscle contraction, mucous secretion and local proteolysis.After initial mastocyte immediate reaction, eosinophilic granulocyte, basophilic leukocyte and lymphocytic inflow took place after 6-24 hour.This late phase responses can cause chronic tissue's inflammation continuing to be exposed in the antigenic tissue.

Think that take off granule and the eosinophilic granulocyte of IgE dependent mast cells are that IgE by the unbalanced overactivity of Th2 cell and the Th2 mediation that causes thus excessively generates (the Abbas et al. that causes gathering of inflammation part, 1991, Nature 383:787-93; Vercelli, 2001, Curr Opin Allergy ClinImmunol 2001,1:61-5).The representative cytokine of known Th2 cell plays a significant role in these reactions such as IL-4, IL-5, IL-10 and IL-13.In addition, cytokine such as the IFN-γ of Th1 mediation and IL-12 it is reported the negative Th2 of adjusting approach.For example, IFN-γ induces isotype to convert IgG2a in the B cell, and the Th2 that IL-12 will set up in some cases replys the Th1 advantage that converts to (Umetsu and DeKruyff, 1997, J Allergy Clin Immunol 100:1-6; Coffman andCarty, 1986, J Immunol 136:949-54; Gavett et al., 1995, J Exp Med182:1527-36).The various kinds of cell transcription factor is such as generation (Lee et al., 2000, the J Exp Med192:105-15 of cytokine in the differentiation of GATA3 and T-bet control Th1 and Th2 cell and these cells; Ting et al., 1996, Nature 384:474-8; Lighvani et al., 2001, Proc NatlAcad Sci USA 98:15137-42; Szabo et al., 2000, Cell 100:655-69).

In many countries, allergic disease such as anaphylaxis, allergic rhinitis, asthma, atopic dermatitis, food anaphylaxis and urticaria are at the population that involves up to 20%, and popularity degree increases (Wuthrich B. day by day, Int.Arch.Allergy Appl.Immunol.90:3-10,1989).

For example, asthma is infringement whole world millions of people's important pneumonopathy.The typical characteristic of asthma is the periodicity airflow limitation and/or causes the hyperresponsiveness to various stimulations of excessive airway narrows.Further feature can comprise airway inflammation, the gentle daoization of eosinophilia.It is believed that the raising of air flue responsiveness is that the inflammatory cascade reaction by complexity causes that this reaction relates to several cell types, comprises T lymphocyte and eosinophilic granulocyte.In allergic asthma, the relative Th1 cytokine of the Th2 cytokine status of having the advantage.

Atopic dermatitis (AD) is chronic and the recurrent inflammatory dermatosis, be characterised in that to itch and the change of eczema dermatoses, and the IgE level raises.The sickness rate of AD is the gesture of rising day by day in global baby and child.AD patient's dermatosis is characterised in that the infiltration of inflammatory cells such as T lymphocyte, monocyte/macrophage, eosinophilic granulocyte and mastocyte.These cells are by discharging pathogenesis and the generation that IL-4, IL-5, IL-10, IL-13, oxyphil cell activate the various kinds of cell factor such as chemotactic factor (eotaxin) and TARC and chemotactic factor participation AD.In many cell types, the Th2 cell that generates IL-4, IL-5, IL-10 and IL-13 is at the interim performance pivotal role of the starting of disease progression (Leung, 1997, Clin Exp Immunol 107 (suppl.1): 25-30).IL-4 and IL-13 serve as the main isotype inducer that converts IgE to, and IL-5 induces eosinophilic granulocyte's activation, and the secretion oxyphil cell activates multiple chemotactic factors such as chemotactic factor.The generation that the IL-10 that is generated by monocyte/macrophage and Th2 cell improves TARC, TARC is a kind of Th2 specificity chemotactic factor, known its crossed in the AD pathological changes and expressed.Although at visible Th1 cytokines in the AD dermatosis in the late period of this disease, such as IFN-γ, yet think that the generation of AD mainly is excessive by the generation of the cytokine/chemotactic factor of Th2 mediation and IgE, and (the Jonathan et al that causes of the generation defective of IFN-γ and IL-12,1999, JClin Invest 103:1103-11; Christian et al., 1999, J Clin Invest 104:1097-105; Tomomi et al., 2001, J Allergy Clin Immunol 107:353-8; Weilie et al., 2002, JClin Invest 109:621-8).Yet, reply unbalance relevant cutter system really with excessive generation of IgE and Th1/Th2 and do not illustrate as yet.

Because there are a large amount of evidences to show that Th1 and Th2 type are reflected at body inner phase intermodulation control, so think that regulation and control Th1/Th2 is the reasonable strategy (Kato et al., 1999, J Immunol162:7470-9) of exploitation Remedies for allergic diseases.For example, to the cytokine receptor antagonist of recombinant cytokine such as IL-12 and IFN-γ or IL-4 and IL-5 tested their regulation and control Th1 and Th2 reply between equilibrated ability (Hofstra et al., 1998, J Immunol 161:5054-60; Tomkinson et al., 2001, J Immunol166:5792-800).Yet, directly use these medicines and usually cause adverse side effect.

Leukotriene is also relevant with multiple inflammation related disease, particularly alterative inflammation.Leukotriene is derived from arachidonic acid, i.e. the precursor of prostaglandin.Leukotriene has two classes.Work in the illness of the main dependence of inflammation therein of first kind neutrophil cell, such as cystic fibrosis, inflammatory bowel and psoriasis.Second class (cysteinyl leukotriene) relates generally to the bronchoconstriction that eosinophilic granulocyte and mastocyte bring out in the asthma.They in conjunction with the high selectivity receptor on bronchial smooth muscle and other airway tissue (O ' Byrne et al., Annals of Internal Medicine 1997; 127:472-80).Know that also leukotriene is important for the pathophysiology of allergic rhinitis, chronic urticaria and atopic dermatitis or eczema.Leukotriene antagonist comprises leukotriene synthetic inhibitor and cysteinyl leukotriene receptor antagonist, can be used for specificity and suppresses vectorial generation of these inflammation or effect.

Reduce the hypothesis that serum IgE level can improve allergic symptoms, obtained using confirmation (the Fahy et al. of the clinical trial of chimeric anti-IgE antibodies (CGP-51901) and recombinant humanized monoclonal antibody (rhuMAB-E25), Am.J.Respir.Crit.Care.Med., 155:1828-1834,1997).Suppress synthetic and excretory diacyl benzimidazole analogues of IgE and bacterial polysaccharides and has been recorded in U.S. Patent No. 6,369 respectively, 091 and U.S. Patent Publication No.20020041885.

Korean patent application No.92-11752 has disclosed the moist medicine of anti-inflammatory, anti-allergy and wind resistance that comprises two flavonoid (biflavonoid), such as the 4 '-O-methyl ochnaflavone that separates from Radix Ophiopogonis (Lonicera japonica), it with the multiple symptom of allergy or inflammation-related in demonstrate therapeutic efficiency.Korean Patent Registration No.100744 has disclosed and has comprised anti-inflammatory, anti-allergy and the radioresistance pharmaceutical composition of several separation from two flavonoidss of Semen Ginkgo (Ginko biloba) leaf.Several tcm prescriptions that comprise Siegesbeckia glabrescens it is reported to have the activity (Kim et al., Phytother.Res., 15:572-576,2001) that reduces IgE.In addition, have been found that many medical herbs are abundant sources of histamine release inhibitors or anti-inflammatory compound.

Other conventional medicine that is used for the treatment of allergic conditions comprises hydryllin, steroid or nonsteroid anti-inflammatory drugs, reaches leukotriene antagonist.Yet these medicines might cause serious adverse, include but not limited to improve to the susceptibility that infects, liver toxicity, drug-induced pneumonopathy, and bone marrow depression.So, this type of medicine is used for the treatment of inflammation clinically, particularly is restricted during alterative inflammation.Using the medicine of antiinflammatory and mitigation symptoms is a serious problem, because they have side effect or them can not attack the basic cause of disease of inflammatory response.Need to continue injury medicine littler, that effect is bigger to treat inflammation.Need so, still that the side effect feature is littler, toxicity is littler and the new product bigger to the specificity of the potential cause of disease of inflammation.

At last, help Th1 type T cell activation, IgG2a generate, and the initiation of the immunne response that generates of relevant Th1 cytokines (for example IFN-γ, IL-6, IL-12, the IL-1) immunne response relevant with alterative inflammation discussed above opposite.That such immunne response can have is intensive, systematic, antitumor and ntiviral characteristic.This area continues the product with this class feature need be provided.

Summary of the invention

One embodiment of the invention relate to the method for the immunne response in the regulation and control mammal.This method comprises that with the amount that is enough to regulate and control the immunne response in the mammal to the hard kiwi fruit prepared product of administration, wherein said hard kiwi fruit prepared product is selected from: fresh fruit (fresh fruit), broken fruit (crushed fruit), the fruit that boiled (boiled fruit), the fruit of cooking (cooked fruit), the fruit of pressing (pressed fruit), the fruit that concentrated (condensed fruit), the dry fruit of crossing (driedfruit), with hard kiwi fruit fruit juice concentrates (hardy kiwifruitjuice concentrate).In this embodiment, described hard kiwi fruit prepared product un-extracted.

Aspect of previous embodiments, described hard kiwi fruit prepared product is by comprising that the method with dry this step of fruit obtains.

Another embodiment of the invention relates to the method for the immunne response in the regulation and control mammal, comprise that with the amount that is enough to regulate and control the immunne response in the mammal to the hard kiwi fruit prepared product of administration, wherein said hard kiwi fruit prepared product is selected from: leaf extract or concentrate, and bark extract or concentrate.

Another embodiment of the present invention relates to the method for the immunne response in the regulation and control mammal.This method comprises with the amount that is enough to regulate and control the immunne response in this mammal administration: (a) hard kiwi fruit prepared product; Reach the composition that (b) is selected from down group: probiotics (probiotics); Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae (turmeric); Curcumin (curcumin); Dimethylsulfone (methylsulfonylmethane) (MSM); Radix Ginseng (ginseng); Rhizoma Zingiberis Recens (ginger); And proanthocyanidin (proanthocyanidin).

In previous embodiments, described hard kiwi fruit prepared product can include but not limited to: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, the fruit that concentrated, the dry fruit of crossing, hard kiwi fruit fruit juice concentrates, in about 80 ℃ water, extract the prepared product that fruit obtains by in temperature being 0 ℃, by directly extracting the prepared product that hard kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that hard kiwi fruit obtains, with by at water, extract the prepared product that hard kiwi fruit obtains in chloroform and the ethyl acetate in turn.

Aspect of previous embodiments, described hard kiwi fruit prepared product is by comprising that the method with dry this step of fruit obtains.In yet another aspect, described hard kiwi fruit prepared product is to extract fruit in about 25 ℃ water and obtain by being about 0 ℃ in temperature.In yet another aspect, described hard kiwi fruit prepared product is to obtain by extract fruit in the water of room temperature.In yet another aspect, described hard kiwi fruit prepared product obtains by directly extracting hard kiwi fruit water-soluble concentrate with ethyl acetate.

Another embodiment of the invention relates to the method for the immunne response of regulation and control in the mammal, comprises with the amount that is enough to regulate and control the immunne response in this mammal administration: (a) hard kiwi fruit prepared product; Reach the composition that (b) is selected from down group: steroid; hydryllin; antibody; antibiotic; cyclosporin (cyclosporins); antifungal (antimycotics); respiratory function regulation agent (respiratory functioncontrollers); analgesic (analgesics); beta-2-agonists (β-agonists); leukotrienes regulator (leukotriene modifiers); cytokine or cytokine receptor antagonist; phosphodiesterase inhibitor; sodium cromoglicate (sodium cromoglycate); nedocromil (nedocrimil); caffeine; theophylline; benzyloxycarbonyl group β-alanyltaurine (carbobenzoxy beta-alanyl taurine); with T cell function inhibitor.

In previous embodiments, described hard kiwi fruit prepared product can include but not limited to: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, spissated fruit, the dry fruit of crossing, hard kiwi fruit fruit juice concentrates, in about 80 ℃ water, extract the prepared product that fruit obtains by in temperature being 0 ℃, by directly extracting the prepared product that hard kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that hard kiwi fruit obtains, with by at water, extract the prepared product that hard kiwi fruit obtains in chloroform and the ethyl acetate in turn.

Aspect of previous embodiments, described hard kiwi fruit prepared product is by comprising that the method with dry this step of fruit obtains.In yet another aspect, described hard kiwi fruit prepared product is to extract fruit in about 25 ℃ water and obtain by being about 0 ℃ in temperature.In yet another aspect, described hard kiwi fruit prepared product is to obtain by extract fruit in the water of room temperature.In yet another aspect, described hard kiwi fruit prepared product obtains by directly extracting hard kiwi fruit water-soluble concentrate with ethyl acetate.

In any embodiment mentioned above, the amount of the kiwi fruit prepared product that is provided is enough to regulate and control Th2 and the Th1 immunne response in the mammal.The amount of the kiwi fruit prepared product that is provided in one aspect, is enough to regulate and control the amount of the antibody isotype that is selected from IgE, IgG2a and IgG1 that mammal generates.The amount of the kiwi fruit prepared product that is provided in yet another aspect, is enough to reduce the generation and/or the level of at least a Th2 cytokine in the mammal or improves the level of at least a Th1 cytokine in the mammal.In yet another aspect, the amount of the described kiwi fruit prepared product that provides is enough to reduce the level or the growing amount of at least a leukotriene in the mammal.The amount of the kiwi fruit prepared product that is provided in yet another aspect, is enough to reduce the expression of the transcription factor that is selected from GATA-3, T-bet and NFATc2 in the mammal.In yet another aspect, the illness that described mammal suffers from or risky generation is such, it is favourable strengthening wherein that Th1 replys and/or suppress that Th2 replys.For example, described mammal can suffer from or risky generation allergic disease or nonallergic inflammatory disease.Described allergic disease can be the disease that is subjected to the leukotriene regulation and control.Described allergic disease includes but not limited to asthma and atopic dermatitis.Again for example, described mammal can suffer from or risky generation viral infection or cancer.

In any embodiment mentioned above, described hard kiwi fruit can include but not limited to: Actinidia arguta Sieb.et Zucc (Actinidia arguta), ovateleaf actinidia leaf (Actinidia kolomikta) and Semen Actinidiae Polygamae (Actinidia polygama), wherein Actinidia arguta Sieb.et Zucc is an embodiment preferred.

In any embodiment mentioned above, described hard kiwi fruit prepared product can provide in compositions, its amount account for by weight described composition total weight about 0.01% and about 95% between.

Aspect of any embodiment mentioned above, step of applying comprises hard kiwi fruit prepared product is applied to mammal with carrier, adjuvant or diluent.In yet another aspect, step of applying comprises hard kiwi fruit prepared product is offered mammal as tablet (tablet), powder (powder), effervescent tablet (effervescenttablet), effervescent powder (effervescent powder), capsule (capsule), liquid (liquid), suspension (suspension), granule (granule) or syrup (syrup).In yet another aspect, step of applying comprises hard kiwi fruit prepared product is offered mammal in health food.Health food includes but not limited to: bakery product (fine bakery wares), bread (bread), volume (rolls) stuffs, breakfast cereals (breakfast cereals), finished cheese (processed cheese), the cheese of undressed mistake (unprocessed cheese), flavoring agent (condiments), milk product (dairy products), pudding (puddings), gelatin jelly (gelatin desserts), soda pop (carbonated drinks), teas beverage (teas), drink dried powder (powdered beverage mixes), finished fish product (processed fishproducts), beverage (fruit-based drinks) based on fruit, beverage (vegetable-based drinks) based on vegetable, chewing gum (chewing gum), hard sugar (hard confectionery), refrigerated milk product (frozen dairy products), finished meat products (processed meat products), coating (nut-based spreads) based on nut, wheaten food (pasta), finished poultry product (processed poultry products), thick gravy and sauce (gravies and sauces), potato chips (potatochips), vegetable chips (vegetable chips), crispy slice (crisps), chocolate (chocolate), cookies (cookies), confection (candy), Liquoride sugar (licorice), ice cream (ice creams), dehydrated food (dehydrated foods), the food of cutting (cut food products), finished food (processedfood products), spice (spices), alcoholic beverage (alcoholic beverages), noodles (noodles), fermented food (fermented foods), soup (soups), powder (soup mixes), bean product (soya basedproducts), coating (vegetable oil-based spreads) based on vegetable oil, with beverage (vegetable-based drinks) based on vegetable.In yet another aspect, step of applying comprises the cosmetic composition that the mammal application is comprised hard kiwi fruit prepared product.Cosmetic composition can provide to include but not limited to following form: beauty treatment emulsion (lotion), cream (cream), essence (essence), skin contraction agent (toner), Emulsion (emulsion), beauty treatment Liniment (pack), soap (soap), shampoo (shampoo), irrigation or hair dye (rinse), cleaning agent (cleanser), bath foam (body washing solution), washing liquid (washing solution) or inorganic agent (treatment).In one aspect, step of applying comprises hard kiwi fruit prepared product is offered mammal in food additive.

Aspect another of any embodiment mentioned above, described method may further include the medicine that administration is selected from down group: fatty acid; Polyketide (polyketides); Organic acid; Little organic compound; Aromatic amino acid; Phenylpropyl alcohol alkyl compound (phenylpropanoids); Terpenoid (terpenoids); Steroid; Alkaloid; Corrin; Porphyrin; The line peptide; Cyclic peptide; Ester peptide (depsipeptides); Amino acid derivativges; Nucleoside; Nucleotide; Carbohydrate; Protein; Cell; Cell debris; The medical herbs prepared product; Spice (spices); Mineral (minerals); Antibacterial (sterilizers); Flavoring agent (seasonings); Vitamin; And electrolyte.

Aspect another of any embodiment mentioned above, described method may further include the medicine that administration is selected from down group: probiotics; Bacteria cell wall and fragment; Whey protein; Taurine (taurine); Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Herba Rosmarini Officinalis (rosemary); Rosmarinic acid (rosemarinic acid); Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; Proanthocyanidin and beta-carotene.Fatty acid includes but not limited to conjugated (conjugated) linolenic acid (linolenic acid), eicosapentaenoic acid (eicosapentaenoic acid), docosahexenoic acid (docosahexaenoic acid), gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid (stearidonic acid).

Aspect another of any embodiment mentioned above, described method further comprises other Actinidia species prepared product of administration.Described other Actinidia species can include but not limited to A.chinensis Planch. (A.chinensis), delicious Fructus actinidiae chinensis (A.deliciosa), Actinidia arguta Sieb.et Zucc, Semen Actinidiae Polygamae and ovateleaf actinidia leaf.

Another embodiment of the present invention relates to the compositions of the immunne response that is used for regulating and control mammal.Said composition comprises the reactive compound of hard kiwi fruit prepared product and at least a other immunne response that is used for regulating and control mammal.In one aspect, described other reactive compound is used for the treatment of or prevents allergic disease in the mammal.In one aspect, described other reactive compound is selected from: steroid, hydryllin, antibody, antibiotic, cyclosporin, antifungal, respiratory function regulation agent, analgesic, beta-2-agonists, leukotrienes regulator, cytokine or cytokine receptor antagonist, phosphodiesterase inhibitor, sodium cromoglicate, nedocromil, caffeine, theophylline, benzyloxycarbonyl group β-alanyltaurine and T cell function inhibitor.In yet another aspect, described other reactive compound is selected from: probiotics; Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; And proanthocyanidin.Fatty acid includes but not limited to conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid.Such compositions can include but not limited to Pharmaceutical composition, health food, food additive or cosmetics.

In compositions mentioned above, described hard kiwi fruit can include but not limited to Actinidia arguta Sieb.et Zucc, ovateleaf actinidia leaf and Semen Actinidiae Polygamae.In one aspect, described hard kiwi fruit prepared product is the extract or the concentrate of the part preparation of group under being selected from of hard kiwi fruit: fruit, leaf, stem, bark, root and combination in any thereof.In yet another aspect, described hard kiwi fruit is selected from: fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing and the spissated fruit of fresh fruit, fragmentation.In yet another aspect, described hard kiwi fruit is exsiccant fruit.In yet another aspect, described hard kiwi fruit prepared product is by comprising that the method with dry this step of fruit obtains.In yet another aspect, described hard kiwi fruit prepared product is hard kiwi fruit fruit juice concentrates.In yet another aspect, described hard kiwi fruit prepared product is to obtain by extract fruit in the water of room temperature.In yet another aspect, described hard kiwi fruit prepared product obtains by directly extracting hard kiwi fruit water-soluble concentrate with ethyl acetate.In yet another aspect, described extract obtains by extract hard kiwi fruit in distilled water.In yet another aspect, described extract is the ethyl acetate extract of hard kiwi fruit.

The medicine that another embodiment of the invention relates to hard kiwi fruit or its prepared product and is selected from down group is used for the treatment of purposes in the compositions with immune dysfunction diseases associated or illness in preparation: steroid; hydryllin; antibody; antibiotic; cyclosporin; antifungal; respiratory function regulation agent; analgesic; beta-2-agonists; leukotrienes regulator; cytokine antagonist; cytokine receptor antagonist; phosphodiesterase inhibitor; sodium cromoglicate; nedocromil; caffeine; theophylline; benzyloxycarbonyl group β-alanyltaurine; with T cell function inhibitor.

Another embodiment of the invention relates to hard kiwi fruit or its prepared product and is selected from down the purposes of the medicine of group: probiotics; Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; And proanthocyanidin.Fatty acid includes but not limited to: conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid.

Aspect of any purposes mentioned above, described compositions can be used for the treatment of and leukotriene generation or active diseases associated or illness.Aspect another of any purposes mentioned above, described disease or illness can include but not limited to atopic dermatitis, asthma, food anaphylaxis, allergic rhinitis and chronic urticaria.Aspect another of any purposes mentioned above, described hard kiwi fruit prepared product can include but not limited to: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, the fruit that concentrated, exsiccant fruit, hard kiwi fruit fruit juice concentrates, by in the water of room temperature, extracting the prepared product that fruit obtains, by directly extracting the prepared product that hard kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that hard kiwi fruit obtains, with by at water, extract the prepared product that obtains in chloroform and the ethyl acetate in turn.

Another embodiment of the present invention relates to the method for the immunne response in the regulation and control mammal.This method comprises that with the amount that is enough to regulate and control the immunne response in the mammal to the common kiwi fruit prepared product of administration, wherein said common kiwi fruit prepared product is selected from: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, the fruit that concentrated, exsiccant fruit, common kiwi fruit fruit juice concentrates, by in the water of room temperature, extracting the prepared product that fruit obtains, by directly extracting the prepared product that common kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that common kiwi fruit obtains, with by at water, extract the prepared product that obtains in chloroform and the ethyl acetate in turn.

The accompanying drawing summary

Fig. 1 has shown the inhibition activity that the various prepared products of Actinidia arguta Sieb.et Zucc generate IgE in the U266B1 cell.The result recently calculates with respect to a percentage of the IgE level that the U266B1 cell of handling with LPS is generated with three independent experiments.

Fig. 2 has shown the dose dependent effect that PG102T and PG102E generate IL-4 in the splenocyte of OVA stimulation.According to IC ₅₀Value determines that the ratio of PG102T and PG102E is alive.

Fig. 3 A-3C has shown PG102T and the effect of PG102E to producing IL-4 or IFN-γ T cell (Fig. 3 A) and producing the number and the interior IgE biosynthesis (Fig. 3 C) of B cell of IgE B cell (Fig. 3 B).Data are meansigma methodss of every kind of cell mass percentage ratio of three independent experiments. ^*, P＜0.05 is than the mice of handling with DW.

Fig. 4 A-4B has shown that PG102T and PG102E to the effect of GATA3, T-bet and NFATc2 expression, analyze by Western trace (Fig. 4 A) and quantitative PCR in real time analysis (Fig. 4 B).Expression of results becomes the meansigma methods ± SEM of three independent experiments. ^*, P＜0.05; ^*, P＜0.01, than the mice of handling with DW, beta-actin and GAPDH are as the application of sample contrast.

Fig. 5 A-5B has shown that PG102T and PG102E to the effect of dermatitis development in the NC mice, use dermatitis index (Fig. 5 A) and scratching incidence rate (Fig. 5 B) to analyze.Numerical value is stated meansigma methods ± SEM of 5-6 animal as. ^*, P＜0.05; ^*, P＜0.01 is than the mice of handling with DW.

Fig. 6 A-6C has shown PG102T and the PG102E influence to the blood plasma level of IgE in the NC mice (Fig. 6 A), IgG1 (Fig. 6 B) and IgG2a (Fig. 6 C).Numerical value is stated meansigma methods ± SEM of 5 animals as. ^*, P＜0.05; ^*, P＜0.01 is than the mice of handling with DW.

Fig. 7 A-7B has shown the influence of the generation (Fig. 7 B) that PG102T and PG102E activate chemotactic factor and TARC to total leukocyte in the peripheral blood and eosinophilic granulocyte's number (Fig. 7 A) and oxyphil cell.Numerical value is stated meansigma methods ± SEM of 5 animals as. ^*, P＜0.05; ^*, P＜0.01 is than the mice of handling with DW.

Fig. 8 A-8B shown PG102T and PG102E in the NC mice from the effect of the skin injury of skin of back (Fig. 8 A) and skin of face (Fig. 8 B). ^*, P＜0.05; ^*, P＜0.01 is than the mice of handling with DW.

Fig. 9 A-9B has shown that PG102T and PG102E activate the influence that chemotactic factor, TARC, GATA3 and pSTAT6 express to IL-4, IL-5, oxyphil cell in the skin injury, measure by ELISA (Fig. 9 A) and Western trace (Fig. 9 B).Numerical value is stated meansigma methods ± SEM of 5 animals as. ^*, P＜0.05; ^*, P＜0.01 is than the mice of handling with DW.The percentage ratio activity of mice is handled in number indication in the bracket with respect to DW.

Figure 10 is the sketch map that is used to produce the process of relatively large PG102T; This refrigerated or otherwise exsiccant kiwi fruit concentrate is also referred to as FD001 (FG refers to food grade carrier).

The influence of the relative extent of mouse boosting cell cellulation factor IL-4, IL-5, IL-10, IL-13 and IFN-γ that the FD001 (PG102T) that Figure 11 has shown 3 kinds of dosage (0.25,1.0 and 10mg/mL) stimulates OVA after 3 days in external exposure.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 10 mices obtains.

The influence of the relative extent of mouse boosting cell cellulation factor IL-4, IL-5, IL-10, IL-13 and IFN-γ that FD001 (PG102T) ethyl acetate (EtOAc) extract that Figure 12 has shown 3 kinds of dosage (0.25,1.0 and 10mg/mL) stimulates OVA after 3 days in external exposure.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 10 mices obtains.

The influence of the relative extent of mouse boosting cell cellulation factor IL-4, IL-5, IL-10, IL-13 and IFN-γ that the Actinidia arguta Sieb.et Zucc fruit juice concentrates that Figure 13 has shown 3 kinds of dosage (0.25,1.0 and 10mg/mL) stimulates OVA after 3 days in external exposure.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 10 mices obtains.

The influence of mouse boosting cell cellulation factor IL-13 that the known inhibitive ability of immunity chemical compound that Figure 14 A and 14B have shown 3 kinds of dosage stimulates OVA after 3 days in external exposure and the relative extent of IFN-γ.The test concentrations of cyclosporin is 0.0083,0.083 and 4.15 μ M, and the test concentrations of dexamethasone is 0.01,0.1 and 1 μ M (Figure 14 A), each some representative meansigma methods of the data that the splenocyte from 10 mices obtains.The test concentrations of Quercetin is 1.0,10 and 25 μ M, each some representative meansigma methods (Figure 14 B) of the data that the splenocyte from 2 mices obtains.Cytokine levels is measured by ELISA.

The influence of mouse boosting cell cellulation factor IL-13 (Figure 15 A) that the water surplus that Figure 15 A and 15B have shown ethyl acetate (EtOAc) extract of FD001 (PG102T), FD001 of 3 kinds of dosage (1.0,3.0 and 10mg/mL) and this process stimulates OVA after 3 days in external exposure and the relative extent of IFN-γ (Fig.15B).Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 8 mices obtains.

Figure 16 A and 16B have shown the influence of the relative extent of the FD001 (PG102T) of 3 kinds of dosage (1.0,3.0 and 10mg/mL) and mouse boosting cell cellulation factor IL-13 (Figure 16 A) that after 3 days OVA is stimulated in external exposure from the powder (for using) of FD001 and IFN-γ (Figure 16 B) in capsule.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 8 mices obtains.

The influence of mouse boosting cell cellulation factor IL-13 (Figure 17 A) that the alternative prepared product that Figure 17 A and 17B have shown Actinidia arguta Sieb.et Zucc stimulates OVA after 3 days in external exposure and the relative extent of IFN-γ (Figure 17 B).FD001 (PG102T), fruit juice concentrates, the extract by the fresh fruit preparation of in water, boiling, and each 3 dosage (1.0,3.0 and 10mg/mL) of the room temperature water extract of fruit have been tested.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 8 mices obtains.

The cytokine of the mouse boosting cell that the alternative plant part prepared product that Figure 18 A and 18B have shown Actinidia arguta Sieb.et Zucc stimulates OVA after 3 days in external exposure generates the influence of the relative extent of IL-13 (Figure 18 A) and IFN-γ (Figure 18 B).The various extracts that fruit juice concentrates, decocting in water bark, root or stem are made, and FD001 tested 3 dosage (1.0,3.0 and 10mg/mL) respectively.Cytokine levels is measured by ELISA.Each some representative meansigma methods of the data that the splenocyte from 8 mices obtains.

Figure 19 A-19C highlights the changes in distribution that takes place between the 1st day and the 14th day among human clinical trial, wherein according to the doctor physician overall evaluation of AD (Physician ' s Global Assessment, PGA) measure (standards of grading are shown in Figure 19 C), the experimenter has positive response to the treatment of atopic dermatitis (AD).Use placebo or 600mg FD001 (PG102T) (being respectively Figure 19 A and 19B) every day for the experimenter, the companion treats with topical steroids.

Detailed Description Of The Invention

The hard Chinese grooseberry of the previous discovery of the inventor, particularly by its some extract/concentrate of deriving, Be commonly referred to as PG102 herein, improve the serum levels of Th1 cell factor and IgG2a, reduce Th2 The serum levels of cell factor and IgE suppresses histamine release of mast cell, and it is anti-to suppress the allergia inflammatory Should, be included in the gentle road of the allergia inflammation hyperresponsiveness mouse model of allergen sensitization and at the rat pawl During oedema is measured (referring to U.S. Patent Publication No.2004/0037909, the same). PG102 is oral activity.

The inventor provides further evidence in this article, namely from tara vine preparation, be called Two kinds of concrete extracts of PG102T and PG102E (being described in detail in hereinafter embodiment 1 and 2) represent Go out the ability that the inhibition that IgE is generated is active and regulate and control selective Th1 and Th2 cell factor. This Regulation activity most possibly is to come real by regulating cell transcription factor GATA-3, T-bet and NFATc2 Existing. The inventor's data point out that PG102T and PG102E have very big potentiality as Th1 and Th2 The innate immunity adjusting control agent of approach is finally as antiabnormal reaction agent. The effect of these extracts is at this paper In in vivo in the model of the allergia inflammation relevant with breathing illness and at the model of atopic dermatitis In be confirmed.

The inventor has also described other hard Chinese grooseberry prepared product in this article, comprise whole fruit, stem, The prepared product of the prepared product of root, bark, new extract, concentrate, fruit juice, drying and un-extracted Prepared product, and proved that this type of hard Chinese grooseberry prepared product is at the energy of its regulation and control TH1 and Th2 cell factor The power aspect has similar characteristic, therefore thinks that they have similar antiallergic action characteristic.

In addition, the inventor has proved that hard Chinese grooseberry prepared product of the present invention reduces the water of leukotriene in vivo Put down and weaken the generation of leukotriene, show that composition of the present invention can be used for treating the disease of leukotriene mediation Disease includes but not limited to atopic dermatitis, asthma, food hypersenstivity, allergic rhinitis and chronic urticaria.

Hard Chinese grooseberry and prepared product thereof are accredited as Th1 and reply reinforcing agent, so that these medicines are controlled being used for Treatment will be benefited from this type of disease of replying and illness (including but not limited to virus infections and cancer) Fang Miante Not attractive.

For the production of total water-soluble extractive of hard Chinese grooseberry and the leaching process of ethyl acetate extract Be recorded in U.S. Patent Publication No.2004/0037909 and embodiments of the invention 1 and 2. In the present invention In the other embodiments, the inventor has found now by alternative extraction, has concentrated or the processing method life The hard Chinese grooseberry prepared product that produces also generates the composition with immunoregulation activity, particularly suppresses to reply anti-The ability that the cell factor of former (for example allergen) generates. This type of alternative prepared product includes but not limited to fruit Juice concentrate, fresh fruit concentrate and the fresh fruit prepared product that boiled.

The inventor also proves in this article: the extract of hard Chinese grooseberry plant part except fruit self Or other prepared product, compare with the water-soluble or ethyl acetate extract of fruit have of equal value or better The immunoregulation activity. For example, the inventor has proved the extraction of hard Chinese grooseberry plant stem, root and bark Thing suppresses the effect from the antigenic stimulus splenocyte cellulation factor of allergen sensitization mouse.

The inventor has also made surprising discovery, i.e. as described in this article hard Chinese grooseberry preparation Thing can serve as the other therapies of various idiocrasy illness, includes but not limited to having based on the therapy of steroids The effect assistant. For example, the inventor finds, the adult who suffers from medium order of severity atopic dermatitis is suffered from The person uses as described in this article powdery tara vine water-soluble extractive, has significantly reduced clinical body The whole grading of the internist who levies. Equally, with use steroids but do not accept the trouble of hard Chinese grooseberry extract The person compares, and is in using together those patients of topical steroids, specific by of patient self assessment Clinical symptoms (rubescent) are significantly alleviated, and notice serious in other clinical symptoms of disease The trend that degree reduces. In small-sized Primary Study, the effect of Chinese grooseberry extract is stopped using the patient No longer remarkable behind the steroids. Therefore, in one embodiment, the present invention relates to described herein Hard Chinese grooseberry prepared product associating (or auxiliary) other therapeutic agent treatment atopic diseases or and immune disorder The purposes of relevant Other diseases. The inventor thinks that composition of the present invention can be used for strengthening other treatment The effect of property and trophism therapy is particularly in idiocrasy illness patient. This enforcement side of the present invention Case is discussed in detail hereinafter.

In yet another embodiment of the present invention, the inventor is surprisingly report in this article, does The process of dry hard Chinese grooseberry before carried no weight, and but was to strengthen the bioactive key factor of hard Chinese grooseberry. In addition, the inventor has only shown in this article that the hard Chinese grooseberry of the drying of getting with the water extraction of room temperature represents Go out the biologically active similar to before being attributed to hard Chinese grooseberry hot water extract or extractive with organic solvent. Also contain Cover the hard Chinese grooseberry of the drying of in cold water or cold water, extracting, so extraction is extended to 0 ℃ and 80 ℃ Between any temperature. The inventor also proposes the hard Chinese grooseberry (example of the drying of un-extracted in this article Dry plate such as hard Chinese grooseberry) can have the biologically active that before had been attributed to hard Chinese grooseberry extract. Therefore, originally Invention relates to the hard Chinese grooseberry of any type of drying, comprises carrying that hard Chinese grooseberry by previous drying obtains Get thing, as the medicine of the immune response in the regulation and control mammal or for the preparation of regulating and control in the mammal The composition of immune response. The more specifically purposes of this medicine or composition has description hereinafter.

In another embodiment, the invention further relates to any one-tenth of Actinidiaceae (Actinidiaceae) The member, any member of Actinidia (Actinidia) particularly, include but not limited to be called Chinese gooseberry or The common Kiwi berry of Kiwifruit is used for providing the usefulness of the Chinese grooseberry composition with immunoregulation activity On the way. In one embodiment, bound by theory not, the inventor thinks other member of Actinidia Dry process can provide dry Actinidia composition, and it is described herein that it has at least a portion The BA that has recognized that of hard Chinese grooseberry. In another embodiment, the inventor thinks Other prepared product of common Chinese grooseberry (for example Chinese gooseberry or Kiwifruit) includes but not limited to The prepared product of any part of fruit, complete fruit, stem, leaf, bark or root comprises that it is any Prepared product or extract comprise dry prepared product, un-extracted but through the prepared product of processing, fresh Fruit, fruit juice or its any extract, concentrate or fraction, can have at least a portion herein The biological characteristics that has recognized that of described hard Chinese grooseberry.

Belong to tara vine, silvervine and the Actinidia kolomicta natural distributed of Actinidiaceae in west primary Leah, NORTH CHINA, Korea S the north and southern. Reported that surpassing 30 kinds belongs to Actinidiaceae Species. Wherein, the fruit called after " Chinese grooseberry " of Chinese gooseberry or Kiwifruit (kiwi), and And be welcome edible fruit. Tara vine and other fruit of belonging to together (silvervine for example And Actinidia kolomicta) as the Chinese medicine of called after " Kiwi berry (mihudo) ", be used for the treatment of hepatopathy, Gastrointestinal disease and urogenital lithiasis and do not have toxicity (Seoul National University Natural Products Science, Tradi-Medi Data Base, dongbang media Co.Ltd.1999). Yet, Before the invention that U.S. Patent Publication No.2004/0037909 puts down in writing, not about using Actinidia Fruit is treated report or the suggestion with prevention of allergic diseases and anallergic inflammatory disease. In addition, exist Before the present invention, not about the extract put down in writing among the U.S. Patent Publication No.2004/0037909 with The report of the effect of outer hard Chinese grooseberry prepared product is not about by in addition firmly unusual of fruit (berry) The report of the effect of the extract of fruit part preparation does not have about each step in the preparation process such as doing Dry importance or hard Chinese grooseberry composition have to atopic diseases or with the immune system imbalance The report of the impact of setting up methods for the treatment of (for example Steroid treatment) effect of other illness of closing.

According to the present invention, mention " hard Chinese grooseberry " and refer to any tara vine, silvervine and dog jujube Kiwi berry, or akin with it, have as described in this article tara vine, a Pueraria lobota jujube Kiwi berry and Actinidia kolomicta biologically active characteristic, particularly the anti-allergy spy about confirming herein Other Kiwi berry of property and immune response/cell factor/leukotriene modulating properties (for example referring to embodiment) Species.

At preparation any composition of the present invention or prepared product, comprise any extract described herein The time, can use any one or a plurality of part of hard Chinese grooseberry or other species of Actinidia, comprise but Be not limited to its fruit (being also referred to as " berry " or " unusual berry " (kiwiberries)), leaf, stem, Bark and root.

Generally mention the concentrated prepared product that extract refers to material (for example hard Chinese grooseberry) herein, usually logical Cross with suitable solvent and therefrom take out composition activated or that want, evaporate then, whole or several All solvents, and adjustment residue or powder process become required standard and obtain. Term " concentrate " refers to remove Gone the material form of its most of basic ingredient or solvent. Therefore, obvious term disclosed herein " extract " in some embodiment can with term " concentrate " Alternate. In external, carry To CE refer in one embodiment by water, lower alcohol (such as methyl alcohol, ethanol etc.), Or its mixture, preferred distilled water or 50-90% ethanol, the more preferably hard Chinese grooseberry system of 70% alcohol extract The hard Chinese grooseberry extract that obtains for thing. The non-polar solven soluble extract that generates thus can pass through Further extract soluble extract with non-polar solven such as hexane, ethyl acetate or dichloromethane solvent And obtain. For the production of the idiographic flow of CE with and estimate and be recorded in U.S. Patent Publication No. 2004/0037909, the same, take in all these type of flow processs as a reference at this. Be used for to follow the tracks of, assessment or Affirmation is according to the other bioassary method of the preferred BA of hard Chinese grooseberry composition of the present invention In this paper embodiment, be described, comprise external and the in vivoassay method.

According to the present invention, mention " PG102T " and refer generally to basically such as U.S. Patent Publication No. 2004/0037909, that put down in writing in the same the embodiment 1 of this announcement (for example referring to) or real such as this paper It is described to execute example 1, total from hard Chinese grooseberry described herein (for example A.argutd) preparation Water-soluble extractive (it also can be described as concentrate in this article). In a preferred embodiment, Total water-soluble extractive is from the tara vine preparation, although for those skilled in the art aobvious and easy What see is from other hard Chinese grooseberry, to include but not limited to the preparation of silvervine and Actinidia kolomicta Total water-soluble extractive of equivalence. If total water-soluble extractive is by large scale process but uses The basic steps substantially the same with the preparation of PG102T (as described in embodiment 3) and producing, that The gained prepared product can be referred to herein as FD001. Mentioning in this article " PG102E " refers to by making The extracting process of knowing with this area routine with chloroform, ethyl acetate and n-butanol to mentioned above The PG102T prepared product carries out separated from solvent (solvent partition) in turn and the ethyl acetate fraction that obtains. For the production of being the extract of PG102T extract and being the concrete side of the extract of PG102E extract Method is described in embodiment 1. In another embodiment of the invention, by directly carrying with ethyl acetate Get FD001 (or PG102T) (namely before the ethyl acetate classification, not having chloroform recovery) and produced not Ethyl acetate extract together. Equally, in a preferred embodiment, ethyl acetate extract is From tara vine preparation, can be from other although it will be apparent to one skilled in the art that Hard Chinese grooseberry includes but not limited to the equivalent extract of silvervine and Actinidia kolomicta preparation.

In some embodiment of the present invention, particularly at conduct processing or preparation extract or concentrate Or a step of other prepared product is when dry with fruit, can from any species production of Actinidia this Extract or concentrate or other prepared product of invention. Therefore, an object of the present invention is to provide and comprise This type of hard Chinese grooseberry, perhaps, if need, the composition of the extract of other species of Actinidia, bag Draw together Pharmaceutical composition, cosmetic composition or can be used as health food goods, health food or beverage, Or food additives (comprising humans and animals (comprising domestic pets) food additives) or make therewith With composition. The such composition intention is used for any method of the present invention, comprises for selective regulation The Th1 of (being in the mammal) and Th2 immune response among the patient are such as being used for by using this type of group Compound prevents or treats allergia and anallergic inflammatory disease or provides antiviral or cancer therapy drug The method of goods or nutrient and healthcare products. Other additive and the composition of composition described herein, with Reach dosed administration and application strategies and be equally applicable to this purpose of the present invention.

When preparing any composition described herein, except extract described herein, bag Draw together above outside specifically described extract/concentrate, the present invention also comprise hard Chinese grooseberry whole fruit, Or through processing but the use of the fruit prepared product of un-extracted, described fruit prepared product includes but not limited to Fresh fruit, broken fruit (dry or fresh), the fruit that boiled are (dry or fresh ), the fruit of cooking, dry fruit, the fruit of pressing, freezing fruit and concentrated fruit Real. Therefore, an object of the present invention is to provide whole fruit or the fruit system that comprises this type of hard Chinese grooseberry The composition of standby thing, comprise Pharmaceutical composition, cosmetic composition or can be used as the health food goods, Health food or beverage or food additives or the composition that uses therewith, described fruit prepared product Can be through the processing of some mode (such as dry, as to boil etc.), but non-must be through extracting. Therefore, In one embodiment, the present invention relates to hard Chinese grooseberry (and the common Chinese grooseberry) preparation of un-extracted Thing. Any such composition intention described herein is used for any method of the present invention, comprise for Th1 and Th2 immune response among the selective regulation patient (being mammal) are such as being used for by executing With such composition prevent or treat allergia and anallergic inflammatory disease or provide antiviral or The method of cancer therapy drug goods or nutrient and healthcare products. Other additive of composition described herein and Composition, and dosed administration and application strategies are equally applicable to this purpose of the present invention.

In addition, when preparing any composition described herein, except extraction described herein Outside the thing, the present invention also comprise by any suitable procedure, from any part of hard Chinese grooseberry produce hard The use of the fruit juice of Chinese grooseberry. Fruit juice can be used as the form of directly producing from fruit (be not diluted or Concentrate) use, fruit juice can dilute, and perhaps fruit juice can concentrate to form fruit juice concentrates. For example, Described in embodiment 3, can make fresh Chinese grooseberry by conventional juice extractor. Juice extractor can remove Remove the pericarp of fruit, generate the mixture of seed, pulp and fruit juice. Can process this mixture then (for example by centrifugal or extruding) separates fruit juice and solid, and can concentrate when needed this fruit Juice (such as by evaporation, distillation, ultrafiltration etc.) provides concentrated fruit juice (being fruit juice concentrates). This The based composition intention is used for any method of the present invention, especially for pre-by using such composition Anti-or treatment allergia and anallergic inflammatory disease or provide antiviral or cancer therapy drug goods or battalion Support the method for health products. Other additive and the composition of composition described herein, and dosage is given Medicine and application strategies are equally applicable to this purpose of the present invention.

In addition, when preparing any composition described herein, as U.S. Patent Publication No. Substituting of the extract of putting down in writing in 2004/0037909, other processing that the present invention includes hard Chinese grooseberry is produced The use of product, described other products includes but not limited to: fruit comprises the room temperature water extraction of dry fruit Get thing; Be lower than the water extract of the hard Chinese grooseberry of carrying out in the water of room temperature in temperature; Root, leaf, stem or The water extract of bark or other extract; Or before extraction, do not have dry, fruit, leaf, stem Or each water extract or concentrate or other extract (fresh fruit of for example extracting in the bark Real). The inventor proposes in this article, can be in scope is 0 ℃ to 80 ℃ the water of any temperature Extract hard Chinese grooseberry, comprise room temperature and lower temperature (for example about 0 ℃ to about 25 ℃).

According to the present invention, generally mention " dry hard Chinese grooseberry " or " dry Chinese grooseberry " comprises (for example common unusual through by any method dry any type of hard Chinese grooseberry or other Chinese grooseberry Really). Term " Chinese grooseberry " can be used for making a general reference any member of Actinidia in this article, comprises above The member of the hard Chinese grooseberry of discussing, and the member that the common Chinese grooseberry of discussion is above arranged equally. Therefore, Dry Chinese grooseberry comprises the part (fruit, leaf, stem, root etc.) of any drying of Chinese grooseberry, bag Draw together the fruit of the fruit of dry complete fruit, dry section, dry fragmentation, dry stripping and slicing Fruit and the dry fruit that concentrates, and any Chinese grooseberry extract, the material that wherein extracts is being carried Get front earlier dry. Extract self need not further dry or processing, although that is normally joined extract The useful form of making composition and be used for storing. Further concentrated extract is excellent for what further use Choosing method includes but not limited to evaporation, distillation, ultrafiltration, counter-infiltration, precipitates, is adsorbed to fixing phase also certainly Fixing phase wash-out and being extracted in the replace solvents. Dried extract or concentrate are excellent for what further use Choosing method includes but not limited to tray drying, spray-drying and freeze drying, and can use also can be dry Auxiliary agent or excipient are such as maltodextrin, microcrystalline cellulose and starch. In one embodiment, excellent The Chinese grooseberry that is selected to drying of the present invention is the Chinese grooseberry prepared product of the drying do not extracted subsequently.

Therefore, composition described herein and method are applicable to the purposes of any hard Chinese grooseberry, comprise Any part of fruit, stem, leaf, bark or the root of hard Chinese grooseberry, or its any extract or concentrated Thing or fraction, any type of complete fruit or process are processed but fruit, the fruit juice of un-extracted, or The purposes of its any extract or concentrate or fraction further comprises hard Chinese grooseberry plant part (plant Part comprises fruit, stem, leaf, bark and/or root) and prepare by the method that comprises drying steps The purposes of plant part prepared product.

Therefore, an object of the present invention is to provide and comprise hard Chinese grooseberry described herein and (namely comprise Any part of fruit, complete fruit, stem, leaf, bark or root comprise its any prepared product Or extract or concentrate, comprise dry prepared product, un-extracted but through the prepared product of processing, new Bright fruit, fruit juice or its any extract, concentrate or fraction), consisting essentially of or by The composition of its composition comprises Pharmaceutical composition, alimentation composition, food additives, health food (bag Draw together beverage or foodstuff) or cosmetic composition. In one embodiment, the invention provides The CE of hard Chinese grooseberry, total water-soluble extractive or ethyl acetate extract. At all situations In, hard Chinese grooseberry prepared product be for selective regulation patient's (being in the mammal) Th1 and The active component of Th2 immune response. Particularly, composition has and is selected from lower group at least a biology Learn active: (a) reduce the number that generates the B cell of IgE among the patient; (b) reducing among the patient (is serum Or in the blood plasma) amount of the IgE that generates; (c) reduce at least a Th2 cell factor (for example IL-4, IL-5, IL-10) generation and/or level; (d) improve at least a Th1 cell factor (for example IL-12, IFN-γ) level; (e) expression of reduction transcription factor GATA-3; (f) improve Transcription Factor T-bet Expression; (g) expression of raising transcription factor NFATc2; (h) generate IgG2a among the increase patient The number of B cell; (i) amount of the IgG2a that generates among the raising patient; (j) improve Th1 T lymphocyte Generation or the activity of (for example CD4+, IFN-γ+) are particularly at inflammation part; (k) reduce Th2 T Generation or the activity of lymphocyte (for example CD4+, IL-4+) are particularly at inflammation part; (l) reduce Generate the number of the B cell of IgG1 among the patient; (m) amount of the IgG1 that generates among the reduction patient; With/ Or (n) reduce level or the generation of at least a leukotriene among the patient.

Compositions as described above can be used for preventing and/or treating wherein to be regulated and control immunne response in mode described herein and will be or estimate to be any disease or the illness useful to the patient.

When being used for this paper, phrase " protection patient avoid disease " refers to the symptom that palliates a disease; Reduce the generation of disease; And/or the order of severity of reduction disease.The protection patient can refer to compositions of the present invention prevent disease after being applied to the patient take place and/or cure or alleviate at least a, preferably surpass the ability of a kind of disease symptoms, sign or the cause of disease.Therefore, the protection patient avoid disease comprise prevent disease (preventative processing) takes place and treat patient's (therapeutic treatment) of suffering from disease with palliate a disease symptom the two.Particularly, thus the protection patient avoids disease or strengthens another kind of therapy obtaining beneficial effect and realizing by the regulation and control specified activity.Beneficial effect can be by those of ordinary skills and/or the trained clinician that treating the patient easily assess.Term " disease " refers to that mammal departs from respect to any of normal health state, comprises the state when having disease symptoms, and departs from (for example infection, gene mutation, genetic defect etc.) and taken place but situation that symptom does not occur as yet.

Generally speaking, medicine described herein, the biologic activity or the biological action that comprise hard kiwi fruit compositions, hard kiwi fruit extract or its any prepared product, be meant in vivo that (promptly in the natural physiological environment of using medicine) or external (promptly under laboratory condition) are measured or observed, by that this medicine represented or that carry out, and returned any function that comes from the natural existence form of this medicine.To the modification of medicine, such as by changing the purification of medicine processing or preparation or medicine, may cause medicine to have and the natural identical biologic activity of medicine that exists, perhaps medicine has and the natural biologic activity that exists medicine to compare and raise or reduce.

Therefore, an object of the present invention is to provide such compositions, comprise Pharmaceutical composition or nourishing healthy (nutraceutical) (nutrition) compositions, it comprises the hard kiwi fruit (any part that promptly comprises fruit described herein, complete fruit, stem, leaf, bark or root, and comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract (can be referred to as active component of the present invention or hard kiwi fruit prepared product of the present invention), the immunne response that is used for regulating and control mammal as active component, the Th2 and/or the Th1 immunne response of saying so more specifically and being used for regulating and control mammal, even the Th1 that is used for strengthening mammal of saying so more specifically replys and/or the Th2 that suppresses in the mammal replys.This active component can be used for treating and/or preventing multiple illness and disease, includes but not limited to allergic disease, nonallergic inflammatory disease, viral infection and cancer.Compositions can further comprise other therapeutic agent or nourishing healthy agent or unite use with it, is used for prevention, treats and/or improves any illness mentioned above or disease.According to the present invention, trophism is used and to be comprised and aim to provide nutrient and nutrient to keep, stablize, strengthen, strengthen or to improve individual health status or organism absorption and use food and the liquid application any of the present invention with organic process of bringing into play function, grow and keeping, comprises the nourishing healthy application.Therapeutic is used and to be comprised and be intended to prevent, treat, handle, recover, alleviate and/or cure the individual any the present invention's application that departs from healthy disease or illness.Other application of the present invention comprises for example cosmetic applications.

It (is hard kiwi fruit prepared product that another object of the present invention provides hard kiwi fruit described herein, the any part that comprises fruit, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, be used to prepare therapeutic agent or nourishing healthy agent, be used for regulating and control the immunne response of mammal, the Th2 and/or the Th1 immunne response of saying so more specifically and being used for regulating and control mammal, even the Th1 that is used for strengthening mammal of saying so more specifically replys and/or the Th2 that suppresses in the mammal replys.This active component can be used for treating and/or preventing multiple illness and disease, includes but not limited to allergic disease, nonallergic inflammatory disease, viral infection and cancer.Medicine can be united use with other therapeutic agent or nourishing healthy agent, is used for prevention, treats and/or improves any illness mentioned above or disease.

Another object of the present invention provides such health food or food additive, it comprises hard kiwi fruit described herein (is hard kiwi fruit prepared product, the any part that comprises fruit, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, and any acceptable additive, be used for regulating and control the immunne response of mammal, the Th2 and/or the Th1 immunne response of saying so more specifically and being used for regulating and control mammal, even the Th1 that is used for strengthening mammal of saying so more specifically replys and/or the Th2 that suppresses in the mammal replys.This type of health food or food supplement can be used for treating and/or preventing multiple illness and disease, include but not limited to allergic disease, nonallergic inflammatory disease, viral infection and cancer.

Another purpose of the present invention provides such animal feed or feed additive, it comprises hard kiwi fruit described herein (is hard kiwi fruit prepared product, the any part that comprises fruit, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, as essential composition, be used for regulating and control the immunne response of mammal, the Th2 and/or the Th1 immunne response of saying so more specifically and being used for regulating and control mammal, even the Th1 that is used for strengthening mammal of saying so more specifically replys and/or the Th2 that suppresses in the mammal replys.This type of animal feed or animal feed additive can be used for treating and/or preventing multiple illness and disease, include but not limited to allergic disease, nonallergic inflammatory disease, viral infection and cancer.

Another purpose of the present invention provides such topical compositions or cosmetic composition, it comprises hard kiwi fruit described herein (is hard kiwi fruit prepared product, the any part that comprises fruit, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, be used for regulating and control the immunne response of mammal, the Th2 and/or the Th1 immunne response of saying so more specifically and being used for regulating and control mammal, even the Th1 that is used for strengthening mammal of saying so more specifically replys and/or the Th2 that suppresses in the mammal replys.This type of cosmetic composition can be used for treating and/or preventing multiple illness and disease, includes but not limited to allergic disease (comprising skin allergy disease or cutaneous allergic disease), nonallergic inflammatory disease, viral infection and cancer.

Any compositions, additive or medicine described herein can comprise carrier, adjuvant or the diluent of at least a routine in addition.For example, compositions of the present invention can comprise materia medica acceptable carrier, adjuvant or diluent, for example lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltose alcohol, starch, Radix Acaciae senegalis, alginate, gelatin, calcium phosphate, calcium silicates, cellulose, methylcellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, nipasol, Talcum, magnesium stearate and mineral oil.Preparaton can comprise filler, anti-agglutinant, lubricant, wetting agent, flavoring agent, emulsifying agent, antiseptic or the like in addition.Compositions of the present invention can be mixed with preparaton, thereby the active component that provides quick after it is applied to the patient, continues or postpones discharges.

For example, compositions of the present invention can be dissolved in oil, propylene glycol or be usually used in generating other solvent of injection.The suitable example of carrier comprises normal saline, Polyethylene Glycol, ethanol, vegetable oil, isopropyl myristate etc., but is not limited to these carriers.For local application, chemical compound of the present invention can be mixed with the form of ointment (ointment) and cream (cream).

Compositions of the present invention or preparaton can be prepared into any form, such as peroral dosage form (effervescent tablet, the effervescent powder, powder, tablet, capsule, soft capsule, liquid medicine (aqueous medicine), syrup, elixir (elixir), pill (pill), wafer (sachet), granule), or topical preparation's (cream, ointment, the beauty treatment emulsion, gel (gel), pomade (balm), patch (patch), paste (paste), spray liquid (spray solution), aerosol (aerosol) or the like), or injectable formulation (solution, suspension, emulsion).

The present composition of pharmaceutical dosage form can use separately, perhaps suitably unite or be used in combination other pharmaceutically active compound, comprise anti-inflammatory chemical compound, antiallergic compounds, maybe can regulate and control immunne response or any other chemical compound or the compositions of benefit are provided for the patient.The chemical compound that special hope is used in compositions of the present invention and preparaton has a detailed description in this article.

Compositions of the present invention can also be as the health food (food of for example various food, beverage, chewing gum (gum), compound vitamin, improvement health or the like) that comprises the hard kiwi fruit prepared product of the present invention.Health food can be used as food, powder, granule, tablet, chewable tablet, capsule or beverage and waits and provide.Child or baby food are also included within the compositions of the present invention, such as the baby or the infant foods of (modified) milk powder that changes composition, infant formula and change composition.

Can include but not limited to bakery product with the suitable food product that generates the health food product to wherein mixing compositions of the present invention or medicine, the bread and the volume that stuffs, breakfast cereals, the cheese of finished and undressed mistake, flavoring agent (tomato catsup, mayonnaise etc.), milk product (breast, yogurt), pudding and gelatin jelly, soda pop, the teas beverage, beverage powder, finished fish product, beverage (comprising fruit juice) based on fruit, beverage (comprising vegetable juice) based on vegetable, chewing gum, hard sugar, refrigerated milk product, finished meat products, nut and based on the coating of nut, wheaten food, finished poultry product, thick gravy and sauce, potato chips and other bits sheet (chips) or crispy slice, chocolate and other confection sweet food class (cookies, confection, Liquoride sugar), ice cream, dehydrated food, cut or finished food (fruit for example, vegetable), spice, alcoholic beverage, noodles, fermented food, soup and powder, bean product (bean milk (milks), the bean beverage, soyabean milk fat (cream), coffee creamer (whiteners)), coating based on vegetable oil, with beverage based on vegetable.Compositions of the present invention can also be used with food, such as being placed on the food when edible, being poured on the food or mixing in food.

Compositions mentioned above, the cosmetic formulations of the compositions that particularly comprises above to be mentioned, can be prepared into any form, such as skin (skin), beauty treatment emulsion, cream, essence, skin contraction agent, Emulsion, beauty treatment Liniment, soap, shampoo, irrigation or hair dye, cleaning agent, bath foam, washing liquid, inorganic agent, gel, pomade, spray liquid or the like.

Any the invention described above compositions can further comprise one or more lactose, casein, dextrose, glucose, sucrose and sorbitol.

Any compositions, prepared product, additive or medicine described herein can comprise at least a activating agent (being reactive compound, active component) in addition.Other activating agent can be pharmacologically active agents and/or nutritional activities agent.Activating agent is compositions contribution at least a extra desirable trophism and/or therapeutic and/or pharmacological characteristics outside the described in this article kiwi fruit prepared product usually.Activating agent can comprise in compositions of the present invention, prepared product, additive or other preparaton with effective dose.The amount of the desired effects that effective dose refers to be enough to realize that this medicine is given is such as to experimenter's health or the effect of nutriture (for example therapeutic or trophism effect), taste effect, abnormal smells from the patient effect, visual effect etc.Those skilled in the art can determine other medicine to be added to the suitable amount in the present composition.

For example, that provided among the present invention or useful any compositions can comprise one or more natural prodcuts as activating agent, includes but not limited to fatty acid and polyketide; Organic acid and little organic compound miscellaneous; Aromatic amino acid and phenylpropyl alcohol alkyl compound; Terpenoid and steroid; Alkaloid; Corrin and porphyrin; Line peptide and cyclic peptide, ester peptide, and other amino acid derivativges; Nucleoside and nucleotide; Carbohydrate; Protein; Cell and cell debris; Medical herbs prepared product and spice; Mineral; Antibacterial; Flavoring agent; Vitamin; Electrolyte; With other natural drug.

Other composition (chemical compound or medicine) that can be added into the present composition comprises any other necessary medicine of synthetic flavoring agent, coloring agent, processing aid, alginic acid or its salt, organic acid, protective colloid binding agent, pH regulator agent, stabilizing agent, antiseptic, glycerol, alcohol, carbonation agent (carbonation agent) or (being used for trophism or therapeutic use by any application process) preparaton, Foods or drinks.

The composition (activating agent) that hard kiwi fruit prepared product preferred especially and of the present invention made up or be added into the compositions that comprises this type of prepared product includes but not limited to: probiotics; Bacteria cell wall and fragment; Whey protein; Taurine; Alanine; Fatty acid (for example conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-, parinaric acid); Monoglyceride, diester and three esters (combination in any of fatty acid constitutes by mentioned earlier); Inositol; Rhizoma Curcumae Longae; Curcumin; Herba Rosmarini Officinalis; Rosmarinic acid; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; Proanthocyanidin; Beta-carotene; Reach and the kiwi fruit kiwi fruit not of the same race that is used as main bioactive ingredients, the any member who comprises Actinidiaceae, any member of Actinidia particularly, comprise common kiwi fruit all kinds of (for example A.chinensis Planch. or delicious Fructus actinidiae chinensis) and hard kiwi fruit all kinds of (for example Actinidia arguta Sieb.et Zucc, Semen Actinidiae Polygamae and ovateleaf actinidia leaf), any other prepared product.

Fatty acid and polyketide include but not limited to: satisfied fatty acid (for example alpha-lipoic acid (R, S or R, S); Unsaturated fatty acid (for example conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-, parinaric acid); Fatty acid ester; Monoglyceride, diglyceride and triglyceride (combination in any of fatty acid constitutes by mentioned earlier); The acetylene series fatty acid; Branched chain fatty acid; Prostaglandin; Thromboxane; Leukotriene; The aromatic series polyketide; Macrolide and polyethers; Lipid-soluble extract (for example marine oils (marine oils), blueweed oil (Echium oil), borage oil (borage oil), olive oil); And lecithin.

Organic acid and little organic compound miscellaneous include but not limited to citric acid; Fumaric acid; 3,8-dimethyl-5-(.alpha.-hydroxyisopropyl)-.DELTA.9-octahydroazulene; Dimethylsulfone (MSM); And ascorbic acid.

Aromatic amino acid and phenylpropyl alcohol alkyl compound include but not limited to aromatic amino acid and benzoic acid (for example benzoic acid, gallic acid (gallic acid), gentisic acid, P-hydroxybenzoic acid, protocatechuic acid, vanillic acid, salicylic acid, syringic acid); Cinnamic acid (for example hydroxytyrosol (hydroxytyrosol), curcumin, rosmarinic acid, α r-turmerone, caffeic acid, eugenol, chlorogenic acid, neochlorogenic acid, cinnamic acid, ferulic acid, o-coumaric acid, P-coumaric acid); Lignan and wood (matter) element; Propenyl benzene; Coumarin; Styryl pyrone (styrylpyrone); (anthocyanidin for example is such as delphinidin (delphinidin) for flavonoid; Proanthocyanidin; Catechin is such as catechin, epicatechin and theaflavin; Flavonol is such as auicularin (avicularin), hyperin (hyperoside), quercimentin (quercitrin), isoquercitrin (isoquercitrin), kaempferol (kaempferol), myricetin (myricetin), rutin (rutin); Flavanone is such as naringenin (naringenin); Chalcone derivative is such as phloretin; Isoflavone is such as vitexin (vitexin)); Stibene (stilbene); Flavanolignan (flavonolignan); Isoflavonoid (isoflavonoid); With terpenoid quinine (terpenoid quinines) (for example vitamin K and tocopherol (vitamin E) are such as tocotrienol (tocotrienol)).

Terpenoid and steroid include but not limited to monoterpene (for example nopinene, Borneolum Syntheticum (borneol), carvacrol (carvacvol), geraniol (geraniol), thymol (thymol), 1,8-cineol (cineol), terpineol (terpineol)); Iridoid (iridoids) (for example monotropein (monotropein)); Alpha, beta-lonone (ionone) (for example 13 carbon precursors of retinoid); Sesquiterpene (for example caryophyllene (caryophyllene), farnesol (farnesol)); Diterpene (for example retinoid); Sesterterpene; Triterpene (for example alpha-amyrin (amyrin), lupeol (lupeol), maloic acid (ursolicacid)); Tetraterpene; Carotenoid (for example lycopene, beta-carotene, phylloxanthin, astaxanthin (astaxanthin), canthaxanthin (canthaxanthin)); And steroid (for example vitamin D class, cupreol).

Alkaloid includes but not limited to pyrrolidine alkaloid, tropane (tropane) alkaloid, two pyrrolidines (pyrrolizidine) alkaloid, piperidine alkaloid, quinolizine alkanes (quinolizidine) alkaloid, indole connection pyridine (indolizidine) alkaloid, pyridine alkaloid, the phenethylamine class, tetrahydroisoquinoline alkaloid, galanthamine (galanthamine), indoles alkaloid, beta-carboline alkaloid, terpenoid indoles (terpenoid indole) alkaloid, quinoline alkaloid, the pyrrolo-indole Alkaloid, Ergota class (ergot) alkaloid, quinazoline alkaloid, quinoline and acridine alkaloid, imidazoles (imizadole) alkaloid, piperidine alkaloid, ephedrine (ephedrine), capsaicin (capsaisin), pyridine monoterpenes alkaloid, aconitine (aconitine), steroid alkaloid, purine alkaloid (allantoin for example, caffeine, theophylline).

Corrin and porphyrin include but not limited to vitamin B.

Line peptide and cyclic peptide, ester peptide, and other amino acid derivativges include but not limited to simple aminoacid and derivant (for example L-acetylcarnitine, choline, taurine, alanine) thereof, line peptide, cyclic peptide (for example cyclosporin), cyclic ester peptide, beta-lactam, cyanogen glycosides (cyanogenic glycoside), glucosinolate (glucosinolate), cysteine sulfoxide (cysteine sulphoxide).

Carbohydrate includes but not limited to monosaccharide (for example inositol), polysaccharide, and (oligofructose for example is such as inulin/inulin (any chain length); Oligomeric galactose; Chitin (chitin) and chitosan (chitosan)).

Other natural materials includes but not limited to protein (for example whey protein and superoxide dismutase); Cell and cell debris (for example probiotics refers to microbial body that live, complete for example some kind of Lactobacillus (Lactobacillus spp.), bacterial cell and cell wall fragments, fungus/yeast cells and cell wall fragments); Medical herbs prepared product and spice (for example Radix Ginseng, huang, Rhizoma Curcumae Longae, Herba Rosmarini Officinalis, Rhizoma Zingiberis Recens); Mineral (for example K, Mg, Ca, Mn, Fe, Cu, Zn, B, Si, Se).The metabolite and the derivant of any of these chemical compound also contain in the present invention.

In one embodiment of the invention, compositions of the present invention is used as the complementary therapy of the routine treatment of illness or disease.For example, the inventor is verified hard kiwi fruit prepared product of the present invention improves the clinical effectiveness of atopic dermatitis patients as the adjuvant of topical steroids therapy the time.Therefore, compositions of the present invention can comprise that one or more are used for the treatment of can be by regulation and control the immunne response illness or the treatment of diseases agent (for example medicine) for the treatment of or alleviating, and they also can be called activating agent in this article.This type of therapeutic agent includes but not limited to that steroid (comprises corticosteroid; comprise oral; suck with injection); hydryllin be (any kind; comprise system; partial; suck, comprise H1 and H2 blocker); antibody (for example anti-IgE; anti-IL-10); antibiotic; cyclosporin; antifungal; respiratory function regulation agent; analgesic; beta-2-agonists (long lasting or fugitive); leukotrienes regulator (inhibitor or receptor antagonist); cytokine or cytokine receptor antagonist; phosphodiesterase inhibitor; sodium cromoglicate; nedocromil; theophylline; caffeine; benzyloxycarbonyl group β-alanyltaurine; T cell function inhibitor and other antiinflammatory.

Any above-mentioned composition can comprise one or more organic acid (citric acid for example in addition, fumaric acid, adipic acid, lactic acid, malic acid, ascorbic acid), phosphoric acid (salt or ester) (phosphoric acid (salt or ester) for example, sodium phosphate, potassium phosphate, acid pyrophosphoric acid (salt or ester), polyphosphoric acid (salt or ester)), and/or the Natural antioxidant (polyphenol for example, catechin, alpha-tocopherol, Herba Rosmarini Officinalis extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.).

Compositions of the present invention, particularly cosmetic composition or preparation are used for topical application of compositions, comprise that therapeutic composition (but being not limited to cosmetics or other topical composition) can comprise other additive, includes but not limited to water soluble vitamins, fatsoluble vitamin, peptide polymer, polysaccharide polymer, sphingolipid, glycosaminoglycans, B-glucosan and seaweed extract.In addition, compositions of the present invention and medicine can be added into existing cosmetics and washing liquid.This based composition can be used as cream, beauty treatment emulsion, massage cream or massage oil, reaches bath foam, soap, shampoo or the like.

Preferred water soluble vitamins be any can with cosmetics or the blended water soluble vitamins of other local preparaton, yet, various vitamin, such as vitamin B 1, B2, B6, pyridoxol (B6), pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotiamide, folic acid, vitamin C, biotin etc., their salt, such as the thiamine salt hydrochlorate, or derivatives thereofs such as SODIUM ASCORBATE such as ascorbic acid-2-phosphonic acids sodium salt, ascorbic acid-2-phosphonic acids magnesium salt is preferred, those water soluble vitamins can pass through conventional method, such as microbe transformation method, method from the culture of microorganism purification, enzyme process or chemical synthesis obtain.

Preferred fatsoluble vitamin be any can with cosmetics or the blended fatsoluble vitamin of other local preparaton, yet, the various vitamin that comprise employed fatsoluble vitamin in the embodiment of the invention are such as vitamin A, D2, D3, E (dl-alpha-tocopherol, the d-alpha-tocopherol, the d-Delta-Tocopherol) and derivant such as ascorbic palmitate, ascorbyl stearate, two ascorbic palmitate, acetic acid-dl-alpha-tocopherol, nicotinic acid-dl-alpha-tocopherol vitamin E, the dl-pantothenylol, the d-pantothenylol, pan-ethyl ether (pantothenyl ethylether) etc. is preferred, those fatsoluble vitamiies can pass through conventional method, such as microbe transformation method, method from the culture of microorganism purification, enzyme process or chemical synthesis obtain.

Preferred peptide polymer be any can with cosmetics or the blended peptide polymer of other local preparaton, yet collagen, hydrolyzable collagen, gelatin, elastin laminin, hydrolyzable gelatin or the keratin etc. that contain by used peptide polymer in the embodiment of the invention are preferred.

Preferred polysaccharide polymer be any can with cosmetics or the blended polysaccharide polymer of other local preparaton, yet hydroxyethyl-cellulose, xanthan gum (xanthin gum), hyaluronate sodium, chondroitin sulfate or their salt (sodium salt etc.) or the like are preferred.For example, can be routinely use from mammal or fish purification of sulphuric acids chrondroitin or its salt.

Preferred sphingolipid be any can with cosmetics or the blended sphingolipid of other local preparaton, yet squalane, ceramide, pit-sphingol (pit-sphingosin), sheath-lipopolysaccharide or the like are preferred.Sphingolipid can use conventional method from separation such as mammal, fish, shellfish (shellfish), yeast or plants and obtain.

Preferred seaweed extract be any can with cosmetics or the blended seaweed extract of other local preparaton, yet, the extract of preferred Brown algae, red algae, chlorella or the like is perhaps from carrageenan, alginic acid, arginic acid, Na, K or the glycosaminoglycans of its isolating purification.Sargassum extract can use conventional method to obtain from the zostera marina purification.

Cosmetics of the present invention and other topical composition can be united other component or be united conventional cosmetics or topical composition, and mentioned above hard kiwi fruit prepared product in case of necessity.Described other component includes but not limited to oil ingredient, wetting agent (humectant), emollient (emollient), surfactant, organic or inorganic dyestuff, organic powder (organic powder), UV absorbent, antiseptic (preservative), antibacterial (antiseptic), antioxidant, plant extract, pH regulator agent, alcohol, pigment, essence (perfume), antipyretic (refrigerant), antiperspirant (antihidrotic), distilled water etc.Preferred oil ingredient can comprise ester oil, hydrocarbon ils, silicone oil, fluoride oil, animal oil, vegetable oil or the like.

Preferred ester oil includes but not limited to three-2 ethyl hexanoic acid glyceride; the 2 ethyl hexanoic acid hexadecyl ester; isopropyl myristate; butyl myristate; isopropyl palmitate; ethyl stearte; octyl palmitate; the different hexadecyl ester of isostearic acid; butyl stearate; Ethyl linoleate; the linoleic acid isopropyl ester; ethyl oleate; the different hexadecyl ester of myristic acid; the different octadecyl ester of myristic acid; the different octadecyl ester of Palmic acid; myristic acid octyl group ten diester; the different hexadecyl ester of isostearic acid; ethyl sebacate; the adipic acid isopropyl ester; neopentanoic acid isoalkyl ester; glycerol three is (sad; capric acid) ester; trimethylolpropane tris-2-ethylhexanoate (trimethylopropanetri-2-ethyl hexanoic acid); trimethylolpropane tris isostearate (trimethylopropanetriisostearic acid); tetramethylolmethane four-2-ethylhexanoate; sad hexadecyl ester; lauric acid ester in the last of the ten Heavenly stems; lauric acid hexyl ester; myristic acid ester in the last of the ten Heavenly stems; myristyl myristate; the myristic acid hexadecyl ester; stearyl stearate; decyl oleate; licinoleic acid ester in the last of the ten Heavenly stems; the different octadecyl ester of lauric acid; different 13 esters of myristic acid; the different hexadecyl ester of Palmic acid; octyl stearate; the different hexadecyl ester of stearic acid; Ceraphyl 140A; oleic acid octyl group ten diester; linoleic acid octyl group ten diester; the isostearic acid isopropyl ester; 2 ethyl hexanoic acid 16 mixed esters; the 2 ethyl hexanoic acid octadecyl ester; the own ester of isostearic acid; glycol dicaprylate; the ethylene glycol bisthioglycolate oleate; propylene glycol dicaprate; propylene glycol two is (sad; capric acid) ester; the propylene glycol dicaprylate; the neopentyl glycol dicaprate; the neopentyl glycol dicaprylate; tricaprylin; glycerol three (undecanoic acid) ester; glycerol three different cetylates; glycerol three isostearates; neopentanoic acid octyl group ten diester; sad different octadecyl ester; different octyl pelargonate; neodecanoic acid hexyl ester in the last of the ten Heavenly stems; neodecanoic acid octyl group ten diester; the different hexadecyl ester of isostearic acid; the different octadecyl ester of isostearic acid; isostearic acid octyl group ester in the last of the ten Heavenly stems; polyglycereol oleanoic acid ester; polyglyceryl-isostearate; citric acid three different hexadecyl esters; citric acid three isoalkyl esters; citric acid three different monooctyl esters; lactic acid ten diester; lactic acid 14 esters; cetyl lactate; Octyldecyl lactate; triethyl citrate; acetyl triethyl citrate; acetyl tributyl citrate; citric acid trioctylphosphine ester; maleic acid two different octadecyl esters; hydroxy stearic acid two 2-Octyl Nitrites; succinic acid 2-Octyl Nitrite; diisobutyl adipate; Dermol DIPS; di-n-octyl sebacate; cholesteryl stearate; the isostearic acid cholesterol ester; the hydroxy stearic acid cholesterol ester; hydroxyl isostearic acid cholesterol ester; Cholesterol 3.beta.-oleate; oleic acid dihydroxy cholesterol ester; isostearic acid pitsteryl ester; oleic acid pitsteryl ester; the different hexadecyl ester of 12-stearoyl hydroxy stearic acid; 12-stearoyl hydroxy stearic acid octadecyl ester; the different octadecyl ester of 12-stearoyl hydroxy stearic acid.

Preferred hydrocarbon ils mentioned above can comprise liquid paraffin (alkane), α-paraffin (olefine) oligomer, different paraffin (alkane), ceresine, paraffin (alkane), the different paraffin of liquid (alkane), polybuden, microwax, vaseline or the like.

Preferred silicone oil can comprise methyl silicone, Methylphenylsilanone., methyl cyclopolysiloxane, prestox polysiloxanes, decamethyl polysiloxanes, ten dimethyl cyclosiloxane, dimethyl siloxane-methyl palmityl oxygen silicone copolymers, dimethyl siloxane-methyl octadecyl oxygen silicone copolymers, alkyl modified silicon oil, amino-modified silicone oil or the like.

Preferred fluoride oil can comprise PFPE or the like.

Preferred animal or vegetable oil can comprise American Avocado Tree oil, almond oil, olive oil, Oleum sesami, corn oil, safflower oil, soybean oil, Semen Maydis oil, Oleum Brassicae campestris, Semen Brassicae Campestris oil, palm-kernel oil, Petiolus Trachycarpi oil, sunflower oil, Oleum Gossypii semen, cocos nucifera oil, cucui nut oil, Semen Tritici aestivi germ oil, the rice germ oil, Adeps Bovis seu Bubali resin, evening primrose oil, macha reaches nurse nut (macadamia nut) oil, herring oil and other fish body oil, egg oil, lanoline, hemp seed oil, ermine oil, orange roughy oil, Simmondsia chinensis oil (jojoba oil), Brazil wax, liquid lanolin or the like.

Preferred humectants can comprise water-soluble low-molecular wetting agent, lipotropy low molecule wetting agent, water-soluble polymer and lipophilic polymer.

Particularly, preferred water-soluble low-molecular wetting agent can comprise cerin(e), glutamine, sorbitol, mannitol, ketopyrrolidine-carboxylic acid sodium, glycerol, propylene glycol, 1,3 butylene glycol, ethylene glycol, Polyethylene Glycol (index of polyphenol＞2), polypropylene glycol (index of polyphenol＞2), lactic acid, lactate or the like.

Preferred fat-soluble low molecule wetting agent can comprise cholesterol, cholesteryl ester or the like.

Preferred water-soluble polymer can comprise CVP Carbopol ETD2050, polyaspartic acid salts, tragacanth, xanthan gum, HMC (hydroxy methocel), HEC (hydroxyethyl-cellulose), HPC (hydroxypropyl cellulose), carboxymethyl cellulose, water soluble chitin, chitosan, dextrin or the like.

Preferred lipophilic polymer can comprise polyvinylpyrrolidone-icosa alkene copolymer, polyvinylpyrrolidone-hexadecylene copolymer, celluloid, dextrin fatty acid ester, silicone polymer or the like.

Preferred emollient can comprise long acyl glutamic acid cholesteryl ester, cholesterol hydroxy stearic acid, 12-hydroxy stearic acid, rogic acid, lanolin fatty acid cholesteryl ester or the like.

Preferred surfactants can comprise nonionic surfactant, anion surfactant, cationic surfactant, amphoteric surfactant or the like.

Particularly, preferred nonionic surfactants can comprise self emulsifying glycerol monostearate, methyl glycol fatty acid ester, fatty acid glyceride, polyglyceryl fatty acid ester, sorbitan aliphatic ester, polyoxyethylene (POE) sorbitan aliphatic ester, POE sorbitan aliphatic ester, POE fatty acid glyceride, POE alkyl ether, POE fatty acid ester, POE solid pimaja oil, POE pimaja oil, POE-POP copolymer, POE-POP alkyl ether, polyether modified silicone, lauric acid alkanol amide, alkyl amine oxide, hydrogenation soybean phospholipid or the like.

The preferred anionic surfactants surfactant can comprise fatty acid soaps; α-acyl group sulfonate; alkylsulfonate; alkyl ally sulfonic acid; alkylnaphthalene sulfonate; alkylsulfonate; the POE alkyl ether sulfate; alkylamide sulfate; alkylphosphonic; the POE alkylphosphonic; alkylamide phosphate; alkyl acyl alkyltaurate (alkyloylalkyl taurine salt); N-acyl group-amino acid salts; the POE alkyl ether carboxy acid salt; alkyl sulfo succinate; alkyl sulfoacetate; acidylate hydrolyzable collagen peptide salt; perfluoralkyl phosphate or the like.

Preferred cationic surfactants can comprise alkyl trimethylammonium chloride, octadecyl trimethyl ammonium chloride, octadecyl trimethyl ammonium bromide, setostearyltrimethyl ammonium chloride, chlorination two (octadecyl) Dimethyl Ammonium, octadecyl dimethyl benzyl ammonium chloride, bromination phenyltrimethyammonium, benzalkonium chloride, diethyl amino yl acetamide stearic acid, dimethylamino propionic acid amide. stearic acid, lanolin derivative quaternary ammonium or the like.

Preferred amphoteric surfactant can comprise carboxybetaine type, amide betaine type, hydroxyl sulfo betaine type, phosphoric acid betaine type, amino carboxylic acid, imidazolidine derivatives type, amide amine type or the like.

Preferred organic and inorganic dyestuff can comprise silicic acid, anhydrous silicic acid, magnesium silicate, Talcum, ceracyte, Muscovitum, Kaolin, colcother (iron sesquioxide), clay, bentonite, titanium film Muscovitum (titanfilm mica), Bismuth Oxychloride (oxy chlorine bismuth), zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminium oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, ferrous oxide, chromium oxide, chromic oxide gel, smithsonite (calamine), white carbon black and combination thereof, as inorganic dyestuff; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl, urea resin, phenol resin, fluoride resin, silicone resin, acrylic resin (acryl resin), melamine resin, epoxy resin, polycarbonate resin, divinylbenzene-styrol copolymer, silk powder (silk powder), cellulose, CI pigment yellow, CI pigment orange are as organic dyestuff; And complex etc.

Preferred organic powder can comprise metallic soap, such as calcium stearate; The alkyl phosphonic acid slaine is such as sodium palmitate zinc, zinc laurate, dodecoic acid calcium; The acylamino acid multivalent metal salt is such as N-lauroyl-Beta-alanine calcium, N-lauroyl-Beta-alanine zinc, N-lauroyl-calcium glycine etc.; The amidosulfonic acid multivalent metal salt is such as N-lauroyl-calcium taurinate, N-palmityl-calcium taurinate; N-acyl group basic amino acid is such as N ε-lauroyl-L-lysine, N ε-palmityl-lysine, N α-palmityl ornithine, N α-lauroyl arginine, sclerosis lanolin fatty acid acyl group arginine or the like; N-acyl group polypeptide is such as N-lauroyl glycyl glycine; The alpha-amido fatty acid,, alpha-amido lauric acid sad or the like such as alpha-amido; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrol copolymer, tetrafluoroethene or the like.

Preferred UV absorbent can comprise para-amino benzoic acid; ethylaminobenzoate; the para-amino benzoic acid pentyl ester; the para-amino benzoic acid monooctyl ester; glycol salicylate; phenyl salicytate; ethylhexyl salicylate; benzyl salicylate; salicylic acid butyl phenyl ester; heliophan (homomentylsalicylate); the benzyl cinnamic acid; to methoxyl group 2-ethoxyethyl group cinnamic acid; to methoxyl group octyl group cinnamic acid; di-p-methoxy single 2-ethyl hexane glyceryl cinnamic acid; to methoxyl group isopropyl cinnamic acid; cinnamic acid diisopropyl-diisopropyl ester admixture; urokanic acid; ethyl urokanic acid; hydroxyl methoxybenzene ketone; hydroxyl methoxybenzene ketone sulfonic acid and salt thereof; dihydroxy methoxybenzene ketone; dihydroxy methoxybenzene ketone sodium disulfonate; the dihydroxy benzenes ketone; the tetrahydroxy benzene ketone; the 4-tert-butyl group-4 '-methoxy dibenzoyl methylmethane; 2; 4; 6-triphen amido-right-(carbonyl-2 '-ethylhexyl-1 '-oxygen)-1; 3, the 5-triazine; 2-(2-hydroxy-5-methyl base phenyl) benzotriazole or the like.

Preferred antiseptic can comprise hinokitiol, three chloric acid, trichloro hydroxyl diphenyl ether, glucuronic acid chlorhexidine, phenyl phenol, resorcin (resorcinol), isopropyl methyl phenol, azulenes (azulene), salicylic acid, zinc pilithione, benzalconium HCl, photosensitizer 301, single nitro guaiaci lignum sodium alkoxide, undecylenic acid or the like.

Preferred anti-oxidants can comprise Butylated hydroxyanisole, Propylgallate, ellisorbate or the like.

Preferred pH regulator agent can comprise citric acid, sodium citrate, malic acid, natrium malicum, fumaric acid, fumaric acid sodium, succinic acid, sodium succinate, sodium hydroxide, sodium hydrogen phosphate or the like.

Preferred alcohol can comprise hexadecanol etc.

In addition, can in any compositions mentioned above, add other component.In one embodiment, the scope of the amount of other component is 0.01 to 5% of a total composition, more preferably 0.01 to 3%.

Component mentioned above, such as water soluble vitamins, fatsoluble vitamin, peptide polymer, polysaccharide polymer, sphingolipid, seaweed extract and other component, can obtain (for example referring to Matsumoto Mithio, Manual for the development of transdermalapplied preparations.Seisi Press, 1st Ed., 1985) by the conventional method that discloses in the document.

It is safe using hard kiwi fruit prepared product of the present invention.They do not have toxicity to animal, and do not represent substantial ill effect.

According to the present invention, any compositions mentioned above can with the hard kiwi fruit prepared product (any part that comprises fruit of the present invention, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its extract or fraction) preparation together, wherein use the hard kiwi fruit prepared product of any suitable dose or appropriate amount, it is enough to realize as mentioned above to the desired biologic activity of hard kiwi fruit prepared product after suitable following period of time is used one or many.Suitable amount or dosage can be with the target of using or illness of being treated or diseases, and with experimenter's body weight, the order of severity, medicament forms, use the path and time-histories changes, and can select by those skilled in the art.

In one embodiment, the suitable amount of the hard kiwi fruit prepared product of the present invention or dosage can comprise about 0.1g to the every kg weight in patients of about 10g, and preferred about 1 to the every kg weight in patients of 3g, as consumption every day of hard kiwi fruit of the present invention or extract of the present invention.Use every day as required potion, every day for several times or the longer time at interval (for example several days once, weekly, every month one inferior).With regard to compositions described herein, the amount of the hard kiwi fruit prepared product of the present invention should be between about 0.01% to 100%, by weight, preferably between about 0.01% to about 95%, by weight, more preferably 0.5 to 80%, by weight, based on composition total weight, comprise any amount between 0.01% and 100%, be amplification with 0.01%.In one embodiment, Pharmaceutical composition of the present invention can comprise the hard kiwi fruit prepared product of the present invention of about 0.01-50% (by weight), based on the gross weight of said composition.

In one embodiment, the extract of the hard kiwi fruit prepared product of the present invention or other prepared product can provide in any compositions with about 20% to 90% highly spissated liquid, powder or granule, comprising any increment between 20% and 90%, is amplification with 1%.

The ratio of other composition usually can be in the scope of about 0 to 20w/w% every 100w/w% compositions in the compositions, comprise 0 and 100w/w% between any increment, be amplification with 1%.

In one embodiment, cosmetic composition with about 0.01 to 30%, more preferably 0.01 to 5% (by weight, gross weight based on compositions) amount comprises hard kiwi fruit prepared product of the present invention, comprises any increment between 0.01% and 30%, is amplification with 0.01%.

In another embodiment, when the compositions that will comprise the hard kiwi fruit prepared product of the present invention is added into food, food additive or beverage, its amount that provides can be in the scope of about 0.1 to 95w/w%, preferred 1 to 80w/w% food, additive or beverage gross weight, comprise 0.1 and 95w/w% between any increment, with 0.1w/w% is amplification, perhaps in the scope of about 1 to 30g every 100ml, preferred 3 to 10g every 100ml health beverage compositionss, comprising any increment between every 100ml of 1g and the every 100ml of 30g, is amplification with 1g.

In one embodiment, health food of the present invention comprises hard kiwi fruit prepared product of the present invention, accounts for 0.01 to 80% of weight, preferred 1 to 50%, based on the gross weight of said composition, comprise any increment between 0.01% and 80%, be amplification with 0.1%.

In one embodiment, () amount comprises hard kiwi fruit prepared product of the present invention to health food drink by weight, based on the gross weight of compositions, comprises any increment between 0.01% and 20%, is amplification with 0.01% with about 0.01 to 20%.Other composition can comprise: aminoacid 0.001 to 5%, by weight, vitamin 0.001 to 2%, by weight, sugar 0.001 to 20%, by weight, organic acid 0.001 to 10%, by weight, the sweeting agent of appropriate amount and spice.As long as health beverage compositions of the present invention comprises the hard kiwi fruit prepared product of the present invention mentioned above as main (essential) composition, then other liquid component is not particularly limited, wherein other composition can be various sweeting agents and/or flavour enhancer, such as adding conventional beverage.The example of this type of sweeting agent or flavour enhancer includes but not limited to sweeting agent conventional or calorie reduction, comprises monosaccharide, such as glucose, fructose etc.; Disaccharide is such as maltose, sucrose etc.; Conventional sugar is such as dextrin, cyclodextrin; And sugar alcohol, such as xylitol, erythritol etc.Other sweeting agent comprises natural sweetener, such as taumatin, Stevia rebaudiana (stevia) extract, levaudioside A, glycyrrhizin and derivant thereof, and calorie sweeting agent that reduces, such as glucide, chlorinated sucrose derivative (sucralose), aspartame and derivant thereof.The amount of sweeting agent mentioned above or flavour enhancer in the scope of about 1 to 20g, preferred 5 to 12g every 100ml beverage composition for treating dental erosion, comprises any increment between the every 100ml of 1g and 20g usually, is amplification with 1g.

Food additive can be added into food by deposition, spraying or mixing.Additive can comprise any increment between 20w/w% and the 100w/w% usually with respect to the amount of total composition in the scope of about 0.01 to 20w/w% every 100w/w% said composition, be amplification with 1w/w%.Food additive can also () amount be mixed such as animal feed with feedstuff by weight, based on the gross dry weight of feedstuff, comprises any increment between the every 1kg of 5g and 100g (by weight), is amplification with 1g to the every 1kg feedstuff of 100g with about 5.

Therefore, an object of the present invention is by using or providing compositions that the Th1 among the selective regulation patient and the method for Th2 immunne response are provided, described compositions comprises Pharmaceutical composition, alimentation health care composition, food additive, health food (comprising beverage or food material), or cosmetic composition, it comprises any hard kiwi fruit prepared product (any part that comprises fruit described herein, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract is made up of it basically as active component, or form by it.Compositions of the present invention has a detailed description hereinbefore.Particularly, use or provide compositions to cause at least a following biologic activity: (a) reduce the number that generates the B cell of IgE among the patient; (b) amount of the IgE that (for example in serum or the blood plasma) is generated among the reduction patient; (c) generation and/or the level of at least a Th2 cytokine of reduction (for example IL-4, IL-5, IL-10); (d) level of at least a Th1 cytokine of raising (for example IL-12, IFN-γ); (e) expression of reduction transcription factor GATA-3; (f) expression of raising Transcription Factor T-bet; (g) expression of raising transcription factor NFATc2; (h) increase the number that generates the B cell of IgG2a among the patient; (i) amount of the IgG2a that generates among the raising patient; (j) improve the generation or the activity of Th1 T lymphocyte (for example CD4+, IFN-γ+), particularly at inflammation part; (k) generation or the activity of reduction Th2T lymphocyte (for example CD4+, IL-4+) are particularly at inflammation part; (l) reduce the number that generates the B cell of IgG1 among the patient; (m) amount of the IgG1 that is generated among the reduction patient; And/or (n) reduce the level or the generation of at least a leukotriene among the patient.

The preferred method of the present invention comprises and reduces that leukotriene generates among the patient, treats thus or alleviates among the patient method with at least a symptom of leukotriene-related illness or disease.This method comprises that any hard kiwi fruit prepared product described herein to described administration effective dose (comprises any part of fruit, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through the prepared product of processing, fresh fruit, fruit juice or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit, total water solubility extract or ethyl acetate extract in one embodiment, with and pharmaceutically acceptable carrier.Preferably, using compositions of the present invention causes the generation of leukotriene among the patient or level to reduce.Include but not limited to asthma, food anaphylaxis, allergic rhinitis, chronic urticaria and allergic dermatitis with leukotriene-related disease and illness.In this embodiment, beyond the systemic application path, preferably use the path and also comprise oral, suction and local application.

Method as described above can be used for preventing and/or treating wherein with mode described herein regulate and control immunne response will be or estimate be to the patient useful any disease or illness.

Therefore, an object of the present invention is to provide the allergic disease that treats and/or prevents in the mammal and the method for nonallergic inflammatory disease, comprise any hard kiwi fruit prepared product (any part that comprises fruit described herein to described administration effective dose, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, use with its pharmaceutically acceptable carrier.

According to the present invention, allergic disease can include but not limited to asthma, allergic bronchopulmonary aspergillosis, allergic bronchitis bronchiectasis, hypersensitivity pneumonitis, allergic sinusitis, anaphylaxis, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact infectious dermatitis, chronic urticaria, insect hypensensitiveness, food anaphylaxis and drug allergy.

In one embodiment, allergic disease is an atopic dermatitis.Aspect of this embodiment, using beyond the compositions that comprises the hard kiwi fruit prepared product of the present invention, forms by it or form basically by it, use the routine treatment of atopic dermatitis simultaneously, include but not limited to the topical steroids medication, treat the patient.In this embodiment of the present invention, most preferably use hard kiwi fruit prepared product by oral or local application, use the path although the invention is not restricted to this type of.

In another embodiment, allergic disease is an asthma.Aspect of this embodiment, using beyond the compositions that comprises the hard kiwi fruit prepared product of the present invention, forms by it or form basically by it, use the routine treatment of asthma simultaneously, include but not limited to topical steroids medication or other asthma controlling agent, treat the patient.In this embodiment of the present invention, most preferably use hard kiwi fruit prepared product by oral or suction, use the path although the invention is not restricted to this type of.

According to the present invention, anallergic scytitis disease can include but not limited to the various skin problems that caused by inflammation, such as pimple, acne etc.As described abovely comprise any hard kiwi fruit prepared product (any part that comprises fruit described herein, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit cosmetic composition in one embodiment, total water solubility extract, or ethyl acetate extract can be used for prevention, treatment and/or improve scytitis among the patient.

Can use compositions described herein and method to prevent or other anallergic inflammation disease for the treatment of includes but not limited to various dermatitis illness, systemic lupus erythematosus (sle) (SLE), retina inflammation, gastritis, retinopathy, hepatitis, enteritis, pancreatitis, nephritis, and wherein to reduce Th2 type immunne response and/or strengthen Th1 type immunne response will be useful similar illness.

Another object of the present invention provides the method that treats and/or prevents the viral infection in the mammal, comprise any hard kiwi fruit prepared product (any part that comprises fruit described herein to described administration effective dose, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract or concentrate, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or concentrate or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, use with its pharmaceutically acceptable carrier.

By the prevention or the treatment viral infection protect mammal at preferred virus include but not limited to Coxsackie virus (Coxsackie virus), cytomegalovirus, epstein-barr virus (Epstein-Barrvirus), banzi virus, hepatitis virus, herpesvirus, influenza virus, Measles virus, rubella virus, human papillomavirus, parainfluenza virus, parvovirus, rabies virus, respiratory syncytial virus, retrovirus and chickenpox virus.

In these viruses, more preferably retrovirus, herpesvirus and hepatitis virus, even more preferably leukemia, (lymphotrophic), sarcoma and the slow virus of having a liking for lymph are as other immunodeficiency or oncovirus.By prevention or treatment viral infection protect mammal at particularly preferredly have a liking for lymph sexually transmitted disease (STD) poison and comprise that T-has a liking for lymph sexually transmitted disease (STD) poison; have a liking for lymph sexually transmitted disease (STD) poison (HTLV is such as HTLV-I and HTLV-II), bovine leukemia virus (BLV) and feline leukaemia virus (FLV) such as people T-cell.Particularly preferred slow virus comprises people (HIV), ape (SIV), cat (FIV) and dog (CIV) immunodeficiency virus, even more preferably HIV-1 and HIV-2.

Another object of the present invention provides the method for cancer that treats and/or prevents in the mammal, comprise any hard kiwi fruit prepared product (any part that comprises fruit described herein to described administration effective dose, complete fruit, stem, leaf, bark or root, comprise its any prepared product or extract, comprise exsiccant prepared product, un-extracted but through processing prepared product, fresh fruit, fruit juice, or its any extract or fraction), be the crude extract of hard kiwi fruit in one embodiment, total water solubility extract, or ethyl acetate extract, use with its pharmaceutically acceptable carrier.

Use the cancer of method and composition of the present invention treatment or prevention include but not limited to melanoma, squamous cell carcinoma, breast carcinoma, head and neck cancer, thyroid carcinoma, soft tissue sarcoma, osteosarcoma, carcinoma of testis, carcinoma of prostate, ovarian cancer, bladder cancer, skin carcinoma, the brain cancer, angiosarcoma (angiosarcomas, hemangiosarcomas), mast cell tumor, primary hepatocarcinoma, pulmonary carcinoma, cancer of pancreas, human primary gastrointestinal cancers, renal cell carcinoma, hemopoietic tumor and metastatic cancer thereof.

Any above-mentioned be used for the treatment of or the method for prevent disease or illness in, can unite the other therapies or the compositions that can be used for treating specific illness and use hard kiwi fruit prepared product.In these embodiments, hard kiwi fruit can be considered as the adjuvant of routine treatment, with the improvement that increases symptom among the patient, recover or alleviate.The conventional medicine that can use with hard kiwi fruit prepared product of the present invention or the special preferred type of therapy include but not limited to that steroid (comprises corticosteroid; comprise oral; suck with injection); hydryllin (any kind, comprise system; partial; suck); antibody (for example anti-IgE; anti-IL-10); antibiotic; cyclosporin; antifungal; respiratory function regulation agent; analgesic; beta-2-agonists (long lasting or fugitive); leukotrienes regulator (inhibitor or receptor antagonist); cytokine or cytokine receptor antagonist; phosphodiesterase inhibitor; sodium cromoglicate; nedocromil; caffeine; theophylline; benzyloxycarbonyl group β-alanyltaurine; T cell function inhibitor and other antiinflammatory.

In the method for the invention, compositions can be applied to or offer any member of vertebrates class (Vertebrate class), mammals (Mammalia) includes but not limited to primates, Rodents, domestic animal, horse and house pet.Preferred patient to be protected is the house pet (for example dog, cat) and the mankind, and is preferred especially human.All mode of administration have been imagined.According to the present invention, term " patient ", " experimenter " and " individuality " are used interchangeably.

Use the path and comprise (in vivo), external (in vitro) and ex vivo (ex vivo) path in the body.Ex vivo refers to the external part regulation and control step of carrying out the patient.The path includes but not limited to that intravenous uses in the body, intraperitoneal is used, intramuscular is used, (intranodal) uses in the joint knot, use in the coronary artery, intra-arterial is used (for example entering carotid artery), subcutaneous administration, percutaneous is delivered, use in the trachea, intraarticular is used, use in the ventricle, suck (for example aerosol), intracranial, in the spinal column, ophthalmic, in ear, intranasal, oral, pulmonary administration, conduit injects (impregnation of a catheter), Intradermal, in the sheath, exterior dura, intracerebral ventricle injection, with be injected directly in the tissue.In one embodiment of the invention, compositions is used (for example subcutaneous, Intradermal, intravenous, intramuscular and intraperitoneal path) by the parenteral path.Intravenous, intraperitoneal, Intradermal, subcutaneous and intramuscular are used and can be used the standard method of this area to carry out.Ear is delivered can comprise [, and intranasal is delivered can comprise nasal drop or nasal injection, and the ophthalmic delivery can comprise eye drop.(referring to for example Stribling et al., Proc.Natl.Acad.Sci.USA 189:11277-11281,1992, in this complete income as a reference) delivered and also can be used the standard method of this area to carry out to aerosol (suction).For example, in one embodiment, compositions of the present invention or vaccine can be mixed with the compositions that is suitable for using suitable suction apparatus or aerosol apparatus to spray and deliver.Oral delivery can for example as tablet or capsule, and be mixed with the Food ﹠ Drink product by with compositions of the present invention and compound the carrying out of carrier that can withstand the Degradation of digestive enzyme in the animal intestinal.The example of examples of such carriers comprises plastic cement wafer or tablet, those that know such as this area.The direct injection technology is useful especially for site-specific ground administered compound.Oral delivery or local delivery are according to the particularly preferred delivery of the present invention or use the path.The path of using of regulation and control mucosal immunity can be used for treating viral infection and some allergia illness.This class.path comprises bronchus, Intradermal, intramuscular, intranasal, other suction, rectum, subcutaneous, local, percutaneous, vagina and urethra path.

Hereinafter the clearer and more definite explanation of embodiment the present invention.Yet, be to be understood that the present invention is limited to these embodiment never in any form.

Embodiment

Embodiment 1

The following examples have proved at least two kinds of specific extraction things from the Actinidia arguta Sieb.et Zucc preparation, are called PG102T and PG102E, contain inhibitory activity that IgE is generated and the ability of regulating and control selectivity Th1 and Th2 cytokine.

Material and method

Mice.BALB/c female mice (6 age in week) derives from Daehan Biolink, and Co.Ltd. (Korea) raises air-conditioning and bioclean indoor and at least 1 week of adaptation are being housed.All experiment flows of hereinafter mentioning all carry out according to the public organizations' animal care and the guide for use (institutional animal careand use guidelines) at Soul national university (Seoul National University) zoopery center (AnimalExperimental Center).

Prepare various extracts from Actinidia arguta Sieb.et Zucc.Employed hard kiwi fruit is available from farm (the Hurstberry Co.Ltd. of this fruit of the special cultivation of a family in this research, Oregon, USA), and (The Moscow Branch of Vavilov Plant Cultivation Research Institute Russia) has confirmed their identity to be indebted to Dr.EllaI.Kolbasina.Exsiccant fruit (10g) is undertaken three times by heating in distilled water (DW) to be extracted.Then it is concentrated, lyophilization, and be dissolved in DW to generate PG102T, concentration is 100mg/ml.The PG102T that will be dissolved in DW extracts in turn with chloroform, ethyl acetate and n-butyl alcohol, obtains PG102C, PG102E and PG102B respectively.Remaining water layer is called PG102W.Each water solublity fraction and last aqueous residue are filtered, concentrate, lyophilization, and with the concentration dissolving of 100mg/ml.All prepared products are stored in-80 ℃ when the needs.

The bioassay of in the U266B1 cell, carrying out.As (Kim et al., Phytother Res 2001 as described in the people such as Kim; 15:572-6) but slightly change, (ATCC, Manassas measure IgE inhibition effect in VA) to the people B lymphoblast oncocyte U266B1 that stimulates at LPS.By people IgEELISA (total people IgE; AlerChek, Portland, Maine) the people IgE level in the detection culture supernatants is by LDH detection kit (Takara Bio, Japan) assessment cell viability.

The in vitro effects that the PG102 pair cell factor of assessing by the anamnestic response in the splenocyte of OVA stimulation generates.Respectively by the 0th day and the 14th day intraperitoneal (i.p.) be injected at 2.25mg aluminium hydroxide (ImjectAlum; Pierce, Rockford, IL) in emulsive 20 μ g ovalbumin (OVA; The V level; Sigma, St.Louis, Mo), with mice (7 age in week) immunity and reinforcement separately subsequently.(naivety) mice of sensitization is not accepted any reagent.At the 24th day, with the OVA sensitization all putting to death (n=5 only/group), separate of the generation of each spleen then with cytokine in the research splenocyte with mice naivety, wherein use as (Yoshimi et al., J Immunol 2000 as described in the people such as Shibata; Anamnestic response 164:1314-21) (recall response).Briefly, isolating splenocyte is seeded to 24 well culture plates, and final concentration is transferred to 5x10 ⁶Individual cell/ml/ hole.Is the OVA of 100 μ g/ml with splenocyte with concentration, and incubation is 3 days under the condition that has PG102T (1mg/ml), PG102C, PG102E, PG102B, PG102W (being 0.1mg/ml) or culture medium in contrast.Behind incubation, collect culture supernatants, use ELISA test kit (Endogen, Cambridge, MA) level of detection cytokine (IL-4, IL-5, IL-12 and IFN-γ).Substantially the same flow process is used to measure the ratio work of PG102T and PG102E.

The measurement of cytokine and immunoglobulin in the OVA sensitization mice.As mentioned above with mouse immune and reinforcement.In order to assess the PG102 prepared product to effect in the body of the inductive allergic response of OVA, mice (n=10/group) with PG102T (15mg/kg/ day) or PG102E (1.5mg/kg/ day) oral processing OVA sensitization, with dexamethasone (DEX, 0.5mg/kg/ day) or DW (100 μ L/ mice/day) is in contrast, from the 14th day to the 24th day once a day.With the inmature mice of the oral processing of DW.At the 21st day, gather blood by the eye blood sampling from each mice, and isolating plasma sample is stored in-80 ℃ when using.(Shibayagi, Gunma Japan) measure total IgE level by mice IgE detection kit.By sandwich ELISA method (Hirano et al., J Immunol Methods 1989; 119:145-50) the level of mensuration total IgG hypotype and the special Ig isotype of OVA.Generate in order to measure cytokine, prepared splenocyte from animal, be resuspended in culture medium (RPMI-1640 that contains 10% FBS), be inoculated into 24 orifice plate (5x10 at the 24th day ⁶Individual cell/ml/ hole) on, and has only the OVA incubation 3 days of 100 μ g/ml with concentration.To cultivate not existing under the OVA condition from the isolating splenocyte of inmature mice.By ELISA (Endogen and R﹠amp; D Systems, Minneapolis, MN) level of IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-γ in the mensuration supernatant.

Immunostaining is analyzed.Splenocyte is exposed to GolgiStop as the intracellular protein transport inhibitors (PharMingen, San Diego, CA) 4 hours, and be ready for detecting and produce the IL-4 cell or produce the IFN-gamma cells.With cell fixation, saturatingization, and with to mice CD4, IL-4 or special PE or the link coupled antibody incubation of PITC of IFN-γ is as (Kyoko et al., J DermScience 2002 as described in the people such as Kyoko; 29:19-25).In order to analyze the IgE product, with cell successively with link coupled anti-mice CD19 of PE and the link coupled anti-mice IgE of FITC (all from PharMingen) incubation.Check cell then, (Becton Dickinson, San Jose CA) analyze the lymphocyte of being established door in the splenocyte to use FACSort analyser and Cell Quest software.For the burnt microscopic analysis of copolymerization, splenocyte is gone up with OVA incubation 2 days at coverslip (cover slip), fixing, saturatingization, and with link coupled anti-mice IgE of FITC and the link coupled anti-mice CD19 dyeing of PE, at last by MRC-1024 laser scanning co-focusing imaging system (Bio-Rad Laboratories Inc., Hercules, CA) observe, as (Semper et al., JAllergy CHn Immunol 2003 as described in the people such as Semper; 112:141-9).

The Western trace.To cultivate 2 days with OVA from each splenocyte of organizing mice, collect (10 ⁷Individual cell/group), then cracking with the preparation protein example.(Santa Cruz Biotechnology, Santa Cruz CA) or as the special antibody of beta-actin (Sigma) of application of sample contrast carry out immunoblotting to mice GATA-3, T-bet, NFATc2 in use.

RNA preparation and quantitative PCR in real time.(GIBCOBRL, Carlsbad CA) separate total RNA from the splenocyte of cultivating 2 days with OVA to use TRIzol reagent.Isolating total RNA is used to use the AMV RT (Roche of system, Mannheim, the Germany) reverse transcription that carries out (RT)-PCR then is to use ABI PRISM 7700 sequence detection systems (Applied Biosystems, Foster City, CA) the quantitative PCR in real time of carrying out.Use the forward and the reverse primer group of the little musculus cdna of Primer Express software (Applied Biosystems) design, nucleotide sequence is as follows:

GATA3

Forward 5 '-CCTCGGCCATTCGTACATG-3 ' (SEQ ID NO:1)

Reverse 5 '-CGTAGTAGGACGGGACGTGG-3 ' (SEQ ID NO:2)

T-bet

Forward 5 '-TGTGGATGTGGTCTTGGTGG-3 ' (SEQ ID NO:3)

Reverse 5 '-ATAAGCGGTTCCCTGGCAT-3 ' (SEQ ID NO:4)

NFATc2

Forward 5 '-GCACATAAGGCCATCAGCTCA-3 ' (SEQ ID NO:5)

Reverse 5 '-TCGCCAGAGAGACTGGCAA-3 ' (SEQ ID NO:6)

GAPDH

Forward 5 '-TGCAGTGGCAAAGTGGAGATT-3 ' (SEQ ID NO:7)

Reverse 5 '-TTGAATTTGCCGTGAGTGGA-3 ' (SEQ ID NO:8)

Respectively organize the difference of the mRNA level of cytogene between mice by Δ Ct value Conversion Calculation.

Statistics.Data are stated meansigma methods ± SEM as, and by the difference between non-matching (unpaired) the Studentt check analysis meansigma methods.Less than 0.05 or 0.01, the P value that calculates as single tail (one-tailed) P value is considered as significant statistically.

The result

The effect of the various prepared products of Actinidia arguta Sieb.et Zucc in the mouse boosting cell that the people U266B1 B cell line and the OVA of LPS stimulation stimulate.The inventor has at first prepared total water solubility extract (PG102T) from Actinidia arguta Sieb.et Zucc, and it is extracted in turn with chloroform (PG102C), ethyl acetate (PG102E), n-butyl alcohol (PG102B) and water (PG102W), obtains four kinds of different fraction of polarity.End user B lymphoblastoma U266B1 cell tests the influences (Fig. 1) that IgE is generated of PG102T and these four kinds of fraction.Find that four kinds of prepared products suppress the U266B1 cell generation IgE that LPS stimulates, except PG102W.PG102E shows the highest inhibitory activity, shows 50% at 25 μ g/ml and suppresses effect (IC ₅₀).PG102T also has activity, its IC ₅₀Value is 126 μ g/ml.All do not demonstrate any cytotoxic effect during all concentration of PG102T and PG102E test in this measures to the U266B1 cell.

In addition, the inventor use the anamnestic response scale-model investigation the various prepared products of Actinidia arguta Sieb.et Zucc to the influence of the generation of the cytokine that relates to Th1 and Th2 approach.Mice was used the OVA immunity at the 0th day, strengthened at the 14th day.After 10 days, mice is put to death, take out spleen with separating Morr. cell.To stimulate with OVA from the splenocyte of OVA sensitization mice, and under the condition of 5 kinds of prepared products that have PG102, cultivated 3 days.As shown in table 1, during cultured cell, the level of IFN-γ, IL-4 and IL-5 is increased to 100 piks or nanogram under the condition that OVA exists.With regard to IL-12, background level is higher, but OVA thorn goad into action this level reduced about 3 times to 415 ± 48pg/ml.PG102T handles and to cause the IL-12 level to raise almost 150%.The performance of IFN-γ is different.From the horizontal detection of the IFN-γ that splenocyte generated of inmature animal less than, but the splenocyte that OVA stimulates has been secreted similar 2.5ng/ml.PG102T handles IFN-γ level is reduced by 40%.Exist at the splenocyte that OVA is stimulated when cultivating under the condition of PG102T, the level of IL-4 and IL-5 has reduced by 62% and 39% respectively.

Table 1:PG102 is to the in vitro effects of the cytokine generation of splenocyte

All splenocytes from OVA sensitization mice stimulate once more with OVA in the training period.Numerical value is stated meansigma methods ± SEM of 5 animals as. ^*, P＜0.05, ^*, P＜0.01 is with splenocyte (StudentT check) contrast of only handling with culture medium, OVA stimulates.ND=detect less than.

The PG102E that demonstrates the most highly inhibited effect that IgE is generated in above-mentioned experiment also reduces by 78% and 59% with the level of IL-4 and IL-5, simultaneously the level of IL-12 is induced 220%.On the contrary, because its cytotoxic effect to splenocyte, PG102C is reduced in the level of all cells factor of measuring in this research.PG102B and PG102W improve the level of IL-12, reduce the level of IFN-γ, IL-4 and IL-5, but be not statistics significantly.In a word, these results show that PG102T and PG102E may contain the chemical compound that suppresses IgE generation and controlled selection Th1 and Th2 cytokine-expressing.Based on above-mentioned data, the inventor selects two kinds of prepared product PG102T and PG102E to be used for research in the further body.

The mensuration that the ratio of PG102T and PG102E is lived., think to contain to surpass a kind of reactive compound to immune polytropism (pleiotropic) activity based on them from the PG102T of plant origin and PG102E.Therefore, in order quantitatively further to test, the inhibitory activity that the IL-4 in the splenocyte that the inventor stimulates OVA based on PG102 as mentioned above generates has been developed reliable bioassary method.With behind the OVA irritation cell, the level of IL-4 by detect less than level be induced to the hundreds of pik.When having PG102T and PG102E, the generation of IL-4 is suppressed (Fig. 2) in dosage dependence mode.All do not show any cytotoxic effect under all concentration conditions of PG102T and PG102E test in this measures.Concentration (the IC of PG102T when 50% inhibitory activity ₅₀) be 806.9 μ g/ml, that PG102E is 91.8 μ g/ml.IC with every kind of prepared product ₅₀Value defined is 1 active unit, and draws total active unit.Use total unit and output calculate PG102T and PG102E when alives, PG102T and PG102E comprise the ratio work of 1.2 units/mg and 10.9 units/mg respectively.This method is used for quality control from the laboratory sample of Actinidia arguta Sieb.et Zucc.

The influence that PG102T and PG102E generate Th1 and Th2 cytokine in the OVA sensitization mouse model.For above vitro data of checking in animal model, use OVA sensitization mouse model test PG102T and PG102E influence to the generation of the various kinds of cell factor that relates to the regulation and control of Th1 and Th2 approach.Mice was used the OVA immunity at the 0th day, strengthened at the 14th day.After the reinforcement, give oral PG102T of animal (15mg/kg/ day=18 unit/kg/ day) or PG102E (1.5mg/kg/ day=16.4 unit/kg/ day), and DEX in contrast (0.5mg/kg/ day) or DW, gave from the 14th day to the 24th day every day.Give oral the feeding of inmature mice of not handling with OVA to DW.The concentration of employed PG102T and PG102E is to have the lowest dose level of maximum activity according to dosage-response preliminary experiment in these experiments.At the 24th day, mice is put to death, separate spleen with the preparation splenocyte.To collect culture supernatants to measure cytokine levels (table 2) from each splenocyte of organizing mice incubation 3 days under the condition that OVA exists.Compare with inmature splenocyte, the level that DW handles IL-12 in the mice reduces.Yet Orally administered PG102T and PG102E improve 1.7 times and 2.6 times with its level respectively.Different with IL-12, OVA stimulates and improves IFN-γ level, and PG102T or PG102E have further improved IFN-γ level.Opposite with PG102T and PG102E, DEX suppresses the two level of IL-12 and IFN-γ.

Effect in the body that table 2:PG102 generates the Th1 of splenocyte and Th2 cytokine

All splenocytes from OVA sensitization mice stimulate once more with OVA.Numerical value is stated meansigma methods ± SEM of 10 animals as. ^*, P＜0.05, ^*, P＜0.01, the mice (Student T check) of handling with DW contrasts.ND=detect less than.

The level of all Th2 cytokines of test raises at the splenocyte camber that OVA stimulates in this research.Yet PG102T handles respectively IL-4, the IL-5 of OVA mediation and the excessive generation of IL-10 has been suppressed 44%, 32% and 44%.PG102E has also suppressed 64%, 69% and 51% with the level of these three kinds of cytokines respectively.DEX has also reduced the concentration of all three kinds of cytokines.PG102 or DEX have reduced IL-13, but be not on the statistics significantly.In a word, these results show that PG102T and PG102E can both controlled selection Th1 and the generation of Th2 cytokine.Almost indistinguishably to suppress all cells factor different with DEX, and PG102T and PG102E show has distinctive biologic activity, can diversity ground regulation and control Th1 and the generation of Th2 cytokine.

PG102T and PG102E are to the influence of the blood plasma level of immunoglobulin isotype.Above the result shows that PG102T and PG102E may reduce the excessive generation of the IgE of Th2 mediation in vivo.Therefore, test oral PG102T and PG102E and handled the blood plasma level that whether can in OVA sensitization mice, control IgE and other immunoglobulin.With the 21st day of the experiment of same type mentioned above, inmature mice generates total IgE of about 140ng/ml, but with the OVA sensitization its level has been improved about 20 times.After handling animal with PG102T and PG102E, the blood plasma level of total IgE has reduced about 2 times.The ability and the DEX of total IgE level are suitable in PG102T or the PG102E downward modulation blood plasma.As seen, use the level that PG102T and PG102E have reduced the IgG1 of Th2 mediation, and the horizontal statistics of the IgG2a of Th1 mediation raises greatly significantly when measuring the level of various IgG hypotypes.The level of IgG2b is not subjected to appreciable impact (table 3A) in all situations.

After the level of measuring special IgE of OVA and IgG hypotype, obtain result (table 3B) much at one.Oral PG102T or PG102E handle and to have reduced special IgE of OVA and the level of IgG1, and the IgG2a that OVA is special has improved above 2 times.These results show that with the data of various Th1 and Th2 cytokine PG102T and PG102E contain adjustable Th1 and Th2 balance, the chemical compound that finally causes rising of IgG2a level and IgE and IgG1 level to reduce.

Table 3:PG102 is to the influence of the blood plasma level of the special immunoglobulin isotype of total and OVA

A.

B.

Numerical value shows meansigma methods ± SEM of 10 animals. ^*, P＜0.05, ^*, P＜0.01 is with the mice contrast of handling with DW (Student T check).Owing to can not get the OVA specific antibody, with the level relatively of antibody in contrast the percentage of OD recently calculate.The OD=optical density.

PG102T and PG102E are to the influence of T in the splenocyte and B cell mass.For understanding the active cell base of PG102T mentioned above and PG102E, the inventor has tested PG102 and has used how to influence T cell and the B cell mass that exists in the splenocyte.From OVA sensitization mice and inmature mice separating Morr. cell, and under the OVA existence condition, cultivated 2 days, then carry out the burnt microscopic analysis of FACS and copolymerization.As shown in Figure 3A, OVA stings the ratio of goading the CD4+IL-4+ cell into action and is increased to 9.3% from 4.7%.PG102T or PG102E use this number are reduced above 30%.On the contrary, when handling animal with PG102T or PG102E, CD4+IFN-γ+cell is increased to 9.2 or 9.6% slightly from 7.2%, although be not on the statistics significantly.The number of two kinds of cell types does not almost change in using the animal of DEX.These results show the ratio that PG102T and PG102E may regulate and control to produce the IL-4T cell and produce IFN-γ T cell.

Then, the inventor has analyzed PG102T and PG102E to producing the influence of IgE B cell.Compare with the DW processing, Orally administered PG102T or PG102E cause the CD19+IgE+B cell number to reduce about 40%.DEX has reduced above 60% (Fig. 3 B) this type of B cell number.The burnt microscopic analysis of copolymerization finds that in addition the level that IgE generates in any given B cell has remarkable reduction.The signal intensity of IgE significantly raises in OVA sensitization mice in the B cell, but reduces greatly after handling animal with PG102T.The effect of PG102E shows the effective force more than PG102T, because the level of fluorescence intensity further reduces.These results show that PG102T and PG102E not only reduce product IgE B cell number, and reduce the intracellular IgE generation of given B.

The influence of PG102T and PG102E pair cell transcription factor.For the biologic activity molecular mechanism behind of understanding PG102T and PG102E, investigated these two kinds of prepared products whether the cell transcription factor that relates to Th1 and Th2 approach has been had any effect.This is because if the direct or indirect regulative transcription factor of PG102 so just can easierly be explained PG102T or the PG102E effect to this type of various kinds of cell factor and IgE.Well-known transcription factor comprises GATA-3, T-bet and NFATc2, the main effect of performance (Lee et al., J Exp Med 2000 in balance Th1/Th2 replys; 192:105-15; Ting etal, Nature 1996; 384:474-8; Lighvani et al., Proc Natl Acad Sci USA 2001; 98:15137-42; Szabo et al., Cell 2000; 100:655-69; Kiani et al., Blood 2001; 98:1480-8).To cultivate 2 days as mentioned above from the isolating splenocyte of OVA sensitization mice, and be ready for immunoblotting assay.The protein level of GATA-3 is reduced greatly by PG102T and PG102E, and the protein level of T-bet and NFATc2 obtains raising (Fig. 4 A).All these transcription factor that the DEX inhibition is tested are as previous report (Adcock, Pulm Pharmacol Ther 2001; 14:211-9).

In order to check this regulation and control to occur in what level, use quantitative real time pcr to measure the mRNA level of each transcription factor (Fig. 4 B).The mRNA level that PG102T handles GATA-3 has reduced almost 3 times, and respectively the mRNA level of T-bet and NFATc2 has been improved about 14 times and 2 times.What is interesting is that though PG102E demonstrates higher biologic activity on the IgE inhibitory action, its degree that changes the level of GATA3, T-bet and NFATc2 is lower than PG102T.DEX reduces the two level of GATA3 and NFATc2, the level of the T-bet that raises slightly.These results show that PG102T and PG102E may be by controlling the generation of Th1/Th2 cytokine in the level of rna level regulative transcription factor such as GATA-3 and T-bet.

Discuss

The biologic activity of viewed PG102T and PG102E shows that strongly the extract of Actinidia arguta Sieb.et Zucc is very suitable for as a kind of potent and unique anti-antiallergic agent in this research.PG102T and PG102E reduce the amount of B cell number and the interior IgE that generates of B cell of product IgE, finally cause blood plasma IgE level to reduce.PG102 seemingly regulates and control Th1 and Th2 balance by level that reduces selectivity Th2 cytokine and the level that improves the Th1 cytokine, thereby has this type of biologic activity.

The influence of PG102T and PG102E pair cell transcription factor may explain how these two kinds of prepared products play a role at molecular level.PG102T and PG102E reduce the GATA-3 level, improve T-bet and NFATc2 level simultaneously.Known GATA-3 strong trans-activation IL-5 promoter and IL-4 enhancer element (Lee et al., J Exp Med 2000; 192:105-15; Ting et al., Nature 1996; 384:474-8).T-bet is by inducing synthetic typing (commitment) (Lighvani et al., the Proc Natl Acad Sci USA 2001 that participates in the Th1 cell of IFN-γ in the Th1 cell; 98:15137-42; Szabo et al., Cell 2000; 100:655-69).In nearest research, reported that also T-bet regulation and control isotype converts generation (Gerth et al., the Int Immunol 2003 of IFN-γ in IgG2a and the B cell to; 15:937-44).The inventor's data are consistent with these known functions of each transcription factor; Promptly the GATA-3 dependency of downward modulation IL-4 and IL-5 is expressed in splenocyte, and raises the T-bet dependency generation of IFN-γ and IgG2a.

Different with T-bet and GATA-3, NFATc2 right and wrong selective expression's the derivable transcription factor of antigen.Although this protein in conjunction with enhancer and the IFN-γ promoter of IL-4, it is believed that NFATc2 becomes in the effect Th1 cell and brings into play more important role driving inmature T cell in the Th of irriate cell.In fact, from NFATc2-/-level of the T emiocytosis IL-4 of mice than wild type T cell much higher (Kiani et al., Blood 2001; 98:1480-8; Erb et al., Infect Immun.2003; 71 (11): 6641-7).This studies show that PG102T or PG102E have improved the NFATc2 expression.

PG102T or PG102E preference Th1 reply and suppress the biosynthetic actual molecules mechanism of IgE still under study for action.A kind of probability is that PG102T or PG102E act on antigen-presenting cell, these cells comprise IL-12 by the secretion soluble factor and express costimulatory molecules such as B7-1 (Th1) and B7-2 (Th2) and in the Th cell differentiation, bring into play pivotal role (Kuchroo et al., Cell 1995; 80:707-18).Another kind of probability is that it directly acts on the atomization of Th CFU-GM or the regulation and control of producing IgE B cell.

PG102T or PG102E may be because the various biological reactive compound to the regulating and controlling effect of Th1 and Th2 system.In order to obtain different PG102 prepared products reliably, based on the influence (IL-4 is the main inducer that the Ig isotype converts IgE to) that they generate IL-4 in the splenocyte that OVA stimulates, the inventor has designed above-mentioned bioassay system.The susceptiveness of this algoscopy and repeatability are enough to calculate the ratio work of PG102 and be used to study the various factors that influences the PG102 biologic activity, such as the diverse geographic location of harvest time and Actinidia arguta Sieb.et Zucc.This quality control that is used for employed PG102 reagent in purification reactive compound and the kinds of experiments based on the algoscopy of IL-4 now.

The inventor's toxicity data shows that PG102T is safe.In the repeated doses toxicity test, Orally administered PG102T (500,1000,2000mg/kg/ day) 4-12 week does not produce harmful effect in rat.These results are with use the data that DEX (0.5mg/kg/ day) obtains to mice opposite fully, and the latter is demonstrating the serious reduction (data not shown) of body weight and spleen quality.These data of describing PG102T and the various biologic activity of PG102E show that the extract of Actinidia arguta Sieb.et Zucc is safe natural prepared product, have minimum side effect risk, might be used for the treatment of multiple allergic conditions.

Embodiment 2

The following examples have proved at least two kinds of specific extraction things from the Actinidia arguta Sieb.et Zucc preparation, are called PG102T and PG102E, have the therapeutic effect to atopic dermatitis.

The NC/Nga mice is to serve as the inbred line that the basis is set up in nineteen fifty-seven with Japan bright and beautiful mice (Nishiki-Nezumi).When raising under specific pathogen-free domestic (SPF) condition, it is normal and healthy that mice keeps.Yet when placing conventional environment, clinical sign at first is the scratching behavior that begins from 8 ages in week, then is the outbreak of eczema disease.The eczema that develops rapidly is usually located at face, ear, neck and back.Ill mice shows the various clinical sign, comprises that hemorrhage, shallow table is rotten to the corn, degree of depth decortication, scurf are peeled off, xerosis cutis and growth retardation (Hiroshi et al., Int Immunol 1997; 9 (3): 461-466).In skin lesion, infiltration and the degranulated mastocyte number of observing a large amount of CD4+T cells and eosinophilic granulocyte increase.In addition, the blood plasma level of IgE significantly raise from 8 ages in week, and is consistent with the appearance of skin lesion.Infiltration cellular expression IL-4, IL-5 and TARC in the skin injury, but IFN-γ expresses and seldom or not to express, and causes showing as the immunoreation that Th2 is a principal mode (Masayuki et al., Int Ach AllergyImmunol 2003; 132:355-63; Christian et al., Mol Med Today 2000; 5:209-10).These discoveries are similar to the inherent various features of AD patient, illustrate that the NC/Nga mice can become the premium animal model for people AD, and regulate and control Th1/Th2 and suppress the IgE biosynthesis to become the therapeutic strategy that can in people and mice, fundamentally improve the AD clinical symptoms.

Material and method

Animal.(SPF) female NC/Nga (NC) mice of specific pathogen free available from SLC (Tokyo, Japan).Mice (5-6 age in week) is raised (SPF NC mice) and autoclaved food and water are provided in the SPF environment, carry out at least one week before use.The SPF NC mice in 7 ages in week is transferred to the conventional room (conventional NC mice) that does not have air-conditioning.University's animal care of Soul national university (SeoulNational University) and the standard of using committee's guide (the University AnimalCare and Use Committee guidelines) to list are abideed by in zoopery.

PG102T or PG102E's is Orally administered.Employed PG102T and PG102E prepare from Actinidia arguta Sieb.et Zucc in this research, as discussed previously (Park et al., J.Allergy Clin.Immunol., 116:1151-1157,2005 and embodiment 1).Conventional NC mice is divided into 3 groups (n=6-8/group), and oral PG102T (50mg/kg/ day), PG102E (5mg/kg/ day), DEX (2.5mg/kg/ day) or distilled water (DW; 100 μ L/ mice/days), from the 7th thoughtful the 14th week once a day.SPF NC mice as negative control is accepted DW.

PG102T or PG102E are to the effect of dermatitis development in the NC/Nga mice.From 7 ages in week age to 14 in week, once in a week by distributing unwitting two people according to people such as Leung (Leung et al., JAllergy Clin Immunol 1990 to handling; 85:927-33) the dermatitis inflammation degree of every group of mice of Ji Zai standard (slightly changing) assessment.Clinical sign of in conventional NC mice, seeing and symptom at first be itch, erythema and hemorrhage, then be that edema, shallow table erosion, degree of depth decortication, scurf peel off, xerosis cutis and growth retardation.To before the skin disease marking,, and assess the index of itching by measuring the scratching time total in 20 minutes to the scratching behavior observation of every mice 20 minutes.The overall clinical order of severity score of AD sample infringement be defined as to 5 S﹠Ss (itch, erythema/hemorrhage, edema, decortication/erosion, scurf peel off/drying) each the grading score (0, nothing; 1, slight; 2, moderate; 3, severe) summation.

The measurement of the blood plasma level of immunoglobulin, cytokine and chemotactic factor.Monitor the allergic response of autonomous induction by the blood plasma level of measuring immunoglobulin (comprising IgE and IgG2a), cytokine and chemotactic factor.Gather blood in 7,10,12 and 14 ages in week with capillary glass tube vascular plexus behind socket of the eye, and isolating plasma sample is stored in-80 ℃ until use.(Shibayagi, Gunma Japan) measure the IgE level, and measure the IgG2a level by sandwich ELISA method, as (Hirano et al., J Immunol Methods 1989 as described in the people such as Hirano with mice IgE detection kit; 119:145-50).Detectable limit is the IgE of 1ng/ml.Also use ELISA test kit (Endogen, Cambridge, MA and R﹠amp; D Systems, Minneapolis MN) has measured the blood plasma level that mice IL-4, IL-12, oxyphil cell activate chemotactic factor (eotaxin) and TARC according to the description of manufacturer.

Th1/Th2 cytokine derived from the splenocyte of NC/Nga mice generates.When 14 ages in week, conventional NC mice is used PG102T, PG102E, DEX or the oral processing of DW, and SPF NC mice is put to death by decapitation.For the cytokine of studying splenocyte generates, obtain spleen from each group, and will be resuspended in culture medium (RPMI-1640 that contains 10% hot deactivation FBS) from the isolating splenocyte of spleen.The splenocyte suspension is seeded to 24 well culture plates, and final concentration is transferred to 5 * 10 ⁶Individual cell/ml/ hole.Then with these cells incubation 3 days when to have or do not exist concentration be the ConA of 2 μ g/ml.The concentration of ConA is optimized according to dosage-response preliminary experiment, and tests employed concentration at this and do not find cytotoxicity.Behind the incubation, collect culture supernatants to measure the level of cytokine (IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-γ) by aforesaid ELISA.

Total leukocyte and eosinophilic granulocyte's analysis in the peripheral blood.When 14 ages in week, gather blood from each group mice.Use Celldyn hematimeter (Abbott; Santa Clara CA) counts total leukocyte in the heparinized blood and eosinophilic granulocyte's number.

The histologic analysis and the measurement of AD sample skin lesion mesocuticle and dermis thickness.For histological examination, obtain the small pieces biopsy samples from the routine in 14 ages in week and head, neck and the skin of back of SPF NC mice.Skin biopsy is fixing in the formalin (pH 7.2) of 10% phosphate-buffered, embedding in paraffin, with 4 μ m section, and with hematoxylin and eosin (H﹠amp; E) dyeing is to detect various inflammatory cells.Observe cell between epitheliums and the sarcolemma at microscopically with 400 times of amplifications.Adjust the telescope to one's eyes after the visual field takes a picture, measure the two thickness of epidermis and corium, be i.e. distance to the basement membrane of corium from the horny layer of epidermis.Distance is expressed as 3 meansigma methodss in the visual field at random, gets measure for 5 times average.

The mensuration of the skin biopsy cells in sample factor and chemokine expression.Measure the level that activates chemotactic factor and TARC from IL-4, IL-5, oxyphil cell in the skin biopsy sample of face by ELISA.In brief, cut out the tissue from the skin of face infringement, homogenate in lysis buffer repeats freezing/thawing flow process 3 times then.After centrifugal, quantitative to the supernatant that contains total cell protein matter, and be used to detect the level of cytokine and chemotactic factor.The result is carried out standardization with respect to the total protein that the self-organizing lysate prepares.Also the molten cytosol sample of aforesaid protein from the skin of face tissue preparation is carried out the Western trace, wherein use (Santa CruzBiotechnology to mice GATA-3, pSTAT6, Santa Cruz is CA) or as the special antibody of beta-actin (Sigma) of application of sample contrast.

Statistics.Data are stated meansigma methods ± SEM as, and by the difference between non-matching (unpaired) the Studentt check analysis meansigma methods.It is significant that the P value that calculates less than 0.05 or 0.01, as single tail (one-tailed) P value is considered as statistics.

The result

Orally administered PG102T and PG102E suppress the development of spontaneous dermatitis in conventional NC mice.In order to check the possible effect of PG102 to atopic dermatitis, the inventor uses the model of NC mice as people's atopic dermatitis, and they demonstrate the skin lesion of atopic dermatitis sample with age growth under normal condition.Give the Orally administered PG102T[50mg of conventional NC mice (60 units)/kg/ day], PG102E[5mg (54.5 units)/kg/ day], DEX (2.5mg/kg/ day) or DW (100 μ L/ mice/day), use every day, continued for 7 weeks, and observe the progress of atopic dermatitis.The dosage of PG102T and PG102E is based on the concentration (referring to embodiment 1 and Park et al., J.Allergy Clin.Immunol., 116:1151-1157,2005) that provides therapeutic effect in the employed OVA sensitization of the previous experiment mouse model.The development that the increase of the dermatitis order of severity score of the conventional NC mice of handling with DW demonstrates dermatitis makes progress (Fig. 5 A) in age dependency mode.Yet Orally administered PG102T or PG102E are from remarkable reduction score in 9 ages in week.The improvement of the dermatitis order of severity is accompanied by the reduction of scratching incidence rate.PG102T handles from 9 ages in week and shortens the scratching time greatly.In the animal of handling, observe similar result (Fig. 5 B) with PG102E.In addition, these results with analyze the observation conclusion consistent (data not shown) that mice overall clinical intuitionistic feature (overall clinical visualfeature) draws.DEX also reduces the dermatitis order of severity and reduces the scratching behavior.These data show that PG102T and PG102E may suppress the dermatitis of autonomous induction in this animal model.

PG102T and PG102E reduce IgE and IgG1 generates, and they improve the IgG2a generation in the blood plasma simultaneously.Except the Clinical symptoms directly perceived of similar people's atopic dermatitis, conventional NC mice also shows that in dermatitis outbreak back the IgE level raises in the blood plasma.Therefore, the blood plasma level that whether can control the IgG2a of the IgE of Th2 mediation and IgG1 and Th1 mediation to Orally administered PG102T or PG102E is tested.In 7 ages in week, give the Orally administered PG102T of animal, PG102E, DEX or DW every day, and when 7,10,12 and 14 ages in week blood sample collection.Under the SPF condition, total the NC mice usually generates the IgE of about 150ng/ml, still, animal is being placed normal condition following time, the IgE level raises gradually with age growth, almost reaches 17 μ g/ml during 14 ages in week.Use PG102T or PG102E from 10 age in week statistics reduce the IgE blood plasma level significantly, the IgE level reduces by 5 times when causing experiment to finish.PG102T and PG102E reduce the effect and the DEX that is used as positive control suitable (Fig. 6 A) of IgE.When 12 ages in week, measure the level of IgG1 (the another kind of Ig classification of Th2 mediation).Under normal condition, the horizontal exceeding 5mg/ml of the IgG1 that the mice of handling with DW generates.Yet, use PG102T and PG102E and respectively its level reduced by 75% and 90%.The level of IgG2a (the Ig classification that belongs to the Th1 mediation) has improved about 180% in the conventional NC mice of handling with PG102T.PG102E also induces the IgG2a level (Fig. 6 B) in the blood plasma.These data show that PG102T and PG102E may be by reducing IgE and IgG1 level and suppressing the dermatitis development by improving the IgG2a level.

The Th1/Th2 cytokine that PG102T or PG102E may regulate and control in blood plasma and the splenocyte generates balance.Above data show that PG102T and PG102E influence Th1 and Th2 cytokine expression.In order on molecule and cellular level, to understand the activity of PG102T and PG102E, when 12 ages in week, measure IL-4 in the blood plasma and the level of IL-12, they represent Th2 and Th1 approach respectively.Compare with the conventional NC mice of handling, reduced by 60% and 76% (table 4A) respectively with IL-4 level in the mice of PG102T or PG102E processing with DW.Orally administered PG102T or PG102E statistics have improved the IL-12 level significantly.

The inventor has also analyzed PG102T and PG102E to separating the effect that Th1 and Th2 cytokine generate in the splenocyte of NC mice.Mice was put to death during ages in 14 weeks, and obtain spleen with separating Morr. cell.To stimulate 3 days with T cell-specific mitogen ConA from each splenocyte of organizing mice, and detect the level of various cytokines.When having ConA, the level of all Th2 cytokines raises greatly, but the level that PG102T or PG102E handle IL-4, IL-5 and IL-10 has reduced by 24% to 78% (table 4B).DEX also suppresses the level of all three kinds of Th2 cytokines, although DEX is not that statistics is significant to the reduction of IL-5.The level of IL-13 is not influenced by PG102 or DEX.

Table 4:PG102T and PG102E are to the influence of Th1 in the splenocyte of blood plasma and cultivation and Th2 cytokine levels

A. blood plasma level

B. edema caused by the spleen disease is flat

Plasma sample separates every group of mice from 12 ages in week.All splenocytes from the NC mice stimulate with ConA in the training period.Numerical value is stated meansigma methods ± SEM of 5 animals as. ^*, P＜0.05, ^*, P＜0.01 is than the mice (Student T check) of handling with DW.ND=detect less than.

When cultivating when having ConA from the cell of conventional NC mice, the level of IL-12 and IFN-γ raises.In SPF NC mice, the level of IL-12 is 166pg/ml, but ConA thorn is goaded it into action and improved 4 times.PG102T or PG102E handle and respectively the IL-12 level have further been induced about 150% or 250%.The level of IFN-γ also significantly is increased to almost 11ng/ml when having ConA.Use PG102T the level of this cytokine has been improved about 150%.PG102E shows the effective force more than PG102T.DEX suppresses the IL-12 level effectively, but it does not influence IFN-γ level.In a word, PG102T and PG102E improve the Th1 cytokine levels, but reduce selectivity Th2 cytokine levels, and DEX is different with them, and it indistinguishably suppresses nearly all tested cytokine expression in this research.

PG102 not only prevents the eosinophilia, and reduces the level that the oxyphil cell activates chemotactic factor and TARC.The skin infiltration of inflammatory cell (comprising the eosinophilic granulocyte) is the key character of atopic dermatitis in the NC mice.Because exist inflammatory cell to be in the skin lesion since they from bone marrow mobilization in blood, total leukocyte and eosinophilic granulocyte's number in the peripheral blood when inventor at first analyzed for 12 ages in week.Shown in Fig. 7 A, the total leukocyte number of conventional NC mice increases in dermatitis outbreak back.Particularly, eosinophilic granulocyte's number increases in the normal condition mice of handling with DW greatly, causes the eosinophilia.Yet, use PG102T or PG102E and reduced the two number of total leukocyte and eosinophilic granulocyte, probably help to prevent increasing of eosinophilic granulocyte.

The variation of eosinophilic granulocyte's number can show as the generation of chemotactic factor in the circulation, and the chemotactic response (chemoattraction) that causes replying inflammation (3-6).Therefore, measure the blood plasma level that the oxyphil cell activates chemotactic factor and TARC (they are representative chemoattractants of eosinophilic granulocyte and Th2 cell).In conventional environment, the mice of handling with DW generates the level rising that the oxyphil cell activates chemotactic factor and TARC, but these levels have reduced about 25% to 50% (Fig. 7 B) in the mice of handling with PG102T or PG102E.Use in the animal of DEX and almost do not change.These results demonstrate PG102T and PG102E suppresses the generation that the oxyphil cell activates chemotactic factor and TARC, cause the prevention of the eosinophilia's (outbreak common and dermatitis takes place simultaneously in the NC mice) to the Th2 mediation.

Use PG102T or PG102E and suppress thickening of inflammatory cell infiltration in the corium and epidermis and corium.PG102T and PG102E also pass through to analyze H﹠amp to the improvement of clinical dermatopathy with to the inhibition that Th2 replys; Painted 14 sections in age in week of E have obtained confirmation.Feed the mice that gives DW shows significant epidermis and corium thickens, give prominence to hyperkeratosis, inflammatory cell infiltration and hemorrhage.Morphology studies show that these infiltration cells in the corium are eosinophilic granulocyte, mastocyte and lymphocyte.Yet, with PG102T or PG102E handle that 7 weeks were suppressed epidermises and corium thickens and corium in the infiltration of inflammatory cell, cause the histology environment (Fig. 8 A) closely similar with SPF NC mice.Use DEX and also produce similar result, but expanded the adipose cell zone greatly.Measurement facial and skin of back epidermis and dermis thickness has proved that also PG102T and PG102E prevent epidermis and corium hypertrophy (Fig. 8 B) significantly with statistics.These results show that oral absorption PG102T or PG102E can effectively suppress the development of dermatitis in the NC mice, and side effect is very little or be free from side effects.

PG102T or PG102E reduce the cytokine of Th2 mediation and the expression of chemotactic factor by downward modulation GATA3.In order to examine or check the effect that PG102T or PG102E generate the cytokine and the chemotactic factor of Th2 mediation in the mouse skin, when 14 ages in week, measure the level that IL-4, IL-5, oxyphil cell activate chemotactic factor and TARC by ELISA.In skin from SPF NC mice, all four kinds of faint expression of protein, but their level significantly raises in conventional NC mice.Using PG102T or PG102E has reduced the level that IL-4, oxyphil cell activate chemotactic factor and TARC above 30%.The effect that IL-5 is expressed is more outstanding, and its level has improved almost 90%.On the contrary, DEX suppresses the level of IL-5 and TARC, but does not suppress the level (Fig. 9 A) that IL-4 and oxyphil cell activate chemotactic factor.

Based on these discoveries, measure the expression of transcription factor (comprising STAT6 and GATA3) by immunoblotting.Well-known STAT6 and GATA3 bring into play pivotal role (Arakawa et al., Clin Exp Immunol 2004 in Th2 cell differentiation and the Th2 specific cell factor and chemotactic factor generation; 135 (3): 505-10; Gunther et al., J Allergy Clin Immunol 2004; 113:987-94; Konishi et al., Proc Natl Acad Sci 2002; 99 (17): 11340-5).Shown in Fig. 9 B, the GATA-3 protein level is subjected to the two reduction of PG102T and PG102E.The expression of phosphorylation STAT6 (pSTAT6) also reduces in the conventional NC mice of handling with PG102T, but the situation of PG102E may be really not so.DEX suppresses the two level of GATA3 and pSTAT6.These results prove that PG102T and PG102E may express the level of reducing the Th2 specific cell factor and chemotactic factor by suppressing GATA3.

Discuss

AD is a kind of main allergic disease, and it usually begins at infancy stage.Quite asthma and/or allergic rhinitis (Leung, Clin Exp Immunol 1997 took place in the diseased individuals of vast scale afterwards; 107 (suppl.1): 25-30).AD is caused by the scytitis that abnormal immune is replied due to (the especially overactivity of Th2 approach).The inventor has found these generations derived from the water solublity fraction of the Actinidia arguta Sieb.et Zucc cytokine of controlled selection Th1 and Th2 mediation in the mouse model of OVA sensitization of PG102T and PG102E, and the generation of IgE (seeing embodiment 1).Based on these data, these plant extracts of reasoning may can be used for treating various allergic diseases (Mayaumi et al., J AllergyClin Immunol 2000; 106:159-66; Hisae et al., Phytother Res 2001; 15:506-10).In this embodiment, the inventor uses the NC/Nga mice as model system, has investigated PG102 and whether can produce therapeutic effect to any reality of atopic dermatitis.The result shows that PG102T and PG102E suppress the development of the dermatitis of autonomous induction, and bound by theory does not think that this is by controlling various Th1 and Th2 correlation factor, promptly reducing IL-4, IL-5, IL-10 and IgE and rise IL-12 and IFN-γ and realize.In the NC/Nga mouse model, these biochemical consequences biology that change comprise the number that significantly reduces eosinophilic granulocyte in peripheral blood and the skin lesion, suppress epidermis and corium and thicken, and suppress the infiltration of various inflammatory cells.The interested especially oxyphil cell of being activates the reduction of the regional expression of chemotactic factor, TARC, IL-4 and IL-5.High unusually in the skin lesion of the NC/Nga mice that these proteinic levels are raised under conventional environment.Yet, after giving the Orally administered PG102 of animal, find that these chemotactic factors and cytokine almost are in normal level.Known oxyphil cell activates chemotactic factor, and IL-5, be eosinophilic granulocyte's strong chemoattractant, and think that the TARC that is produced by keratinocyte (also having the Th2 cell) attracts the Th2 cell, and induce usually the pathologic relevant to reply with atopic dermatitis.In this, it should be noted that the receptor that the oxyphil cell activates chemotactic factor and TARC is respectively CCR3 and CCR4, they all express (Christianet al., J Clin Invest 1999 at Th2 cell camber; 104:1097-105; Tomomi et al., J Allergy Clin Immunol2110; 107:353-8; Weilie et al., J Clin Invest 2002; 109:621-8; Masayuki et al., IntAch Allergy Immunol 2003; 132:355-63).

The therapeutic effect of observed PG102 detailed molecular mechanism behind is with accurately in proper order still in investigation in the NC/Nga mouse model.The possible PG102 of being at first acts on cell transcription factor, GATA-3 for example, relate to the expression of key cytokines such as the IL-4 and the IFN-γ of Th1 and Th2 system subsequently, induce cascade reaction, cause the level of IgE and various chemotactic factors to reduce (Zhu et al., NatImmunol.2004; 11:1157-65).Perhaps, PG102 may at first play a role in local horizontal; For example, reduce the level that the oxyphil cell activates chemotactic factor and TARC, suppress their chemical attraction function, reduce eosinophilic granulocyte's number, and suppress to be found in the histopathology development in the conventional NC/Nga mice of raising at whole body and local horizontal.The activity of PG102 may be owing to a plurality of levels of multiple compound effects in the relevant biochemical route of allergy.In a word, PG102 shows and act on the crucial biological factor that relates to atopic dermatitis in this animal model, treats this illness in its source rather than alleviating of symptom only is provided.

PG102T and PG102E are derived from edible fruit.In the repeated doses toxicity test, do not find toxicity, wherein continued for 12 every days in week and use 2000mg/kg/ day, higher 40 times than employed concentration in this research.In conjunction with the data of the previous experiment that involves OVA sensitization mice, proved that from the result of NC/Nga mice PG102 is used for the treatment of the safe and efficient medicine that various allergic diseases comprise atopic dermatitis.The popularity degree of considering atopic diseases increases day by day in all main developed countries and does not almost have medicine to can be used for the essence treatment of atopic dermatitis, and all those prepared products have as described in this article been represented the progress of this area.

Embodiment 3

The following examples have shown hereinafter the employed preparation that comprises the various prepared products of Actinidia arguta Sieb.et Zucc among the embodiment.

Vegetable material

Actinidia arguta Sieb.et Zucc (Actinidia arguta) (Sieb.Et Zucc.) Planch, ex Miq. (Actinidiaceae) variety ' stem (brachyplast that comprises withe and result), root and the bark collection of Ananasnaya ' from Hurst Berry farm (Sheridan, OR).(CO) the collection manager Mr.TimHogan of herbarium identifies for TheUniversity of Colorado, Boulder, and is deposited in the there by University of Colorado in voucher specimen (#518640).With air-dry 48 hours of vegetable material, extract or other processing before be stored in room temperature.

Sophisticated, edible Actinidia arguta Sieb.et Zucc fruit is gathered from Hurst Berry farm, and is freezing immediately, shipping, freezing preservation (20 ℃) before extraction or other processing.

Extract and other prepared product

Each personal distilled water (1L) of bark (126.2g) of atomizing stem (126.6g), atomizing (79.0g) and in small, broken bitsization was extracted 4 hours in 94 ℃.Then mixture is filtered, and filter liquor is concentrated into drying, obtain stem extract (9.9g), root extract (8.6g) and bark extract (2.4g) by rotary evaporation.

20 pieces of refrigerated Actinidia arguta Sieb.et Zucc berries (154.4g) are melted in room temperature, fragmentation, and with distilled water (1L) in 91 ℃ of extractions 5 hours.Mixture is filtered, and filter liquor is concentrated, obtain " boiling " fresh fruit extract (12.8g).

Other FF kiwi fruit (341.6g) is melted, and juice extractor is passed through in operation.Juice extractor is removed the peel of fruit, generates the mixture of seed, sarcocarp and fruit juice.With this mixture centrifugal (30min, about 3500rpm), obtain 150mL fruit juice.This fruit juice is concentrated into drying by rotary evaporation, obtains fruit juice concentrates (24.2g).

For produce bigger quantity with the suitable extract of PG102T (as described in example 1 above), the kiwi fruit that has carried out process scale extract (Sungil Bioex Co., Ltd., Bibong, Korea).With refrigerated kiwi fruit (1242kg) section (thickness is 1/4 " to 3/8 "), and in countercurrent drier (65-80 ℃), be dried to water capacity 5-20%.In having the jacket type stainless steel reactor of internal screen, exsiccant fruit (239kg) is extracted (Figure 10) in batches with support extraction load.Employing external condensation device prevents the water loss in the leaching process.The quantity of extraction solvent (water) is based on 5-10 times of the weight of exsiccant fruit to be extracted.By in the chuck of reactor, feeding steam, the content of extraction vessel is heated to 90 ℃ from 0 in 2 hours time.Use the outer loop ring to make water (90 ℃) then by biomass circulation 4-12 hour.Subsequently, take out useless biomass and handle, and water extract is filtered by 10 microns filters.Then filter liquor is being concentrated in 55-65 ℃ of stirring-type stainless steel reactor that is being equipped with external condensation device and distillate receptor under the vacuum (about 600mmHg).In case material is concentrated, it is kept 30 minutes so that extract is sterilized again in 80 ℃.Gained material (101kg) that will be suitable with PG102T is called FD001.This material of reserving 3kg is used for further test.In whole process, use Good Manufacturing Practice (Good Manufacturing Practices).

In order to produce the efflorescence material that is suitable for encapsulation and can be used for clinical practice described herein, the FD001 concentrate of above producing (98kg) is evacuated in the horizontal oar agitator and with the microcrystalline Cellulose (MCC) that waits weight (based on the solids content of calculating) with pump mixes.After this, solid mixture is transferred in the stainless steel disc, in forced air exsiccator (70-80 ℃), placed 24 hours.Then the in bulk solid of doing is ground in Fitzmill set hammer formula pulverizer to produce 40 purpose powder (118kg).(Anaheim CA) becomes in the capsule of 300mg or 600mg size for GMPLaboratories of America, Inc., and every FD001 and MCC mixture that contains 1: 1 is used for dog and people's clinical trial with this material encapsulation.

Get cut into slices like that in the initial step as mentioned and dry (but extracting) in batches, from the dry Actinidia arguta Sieb.et Zucc fruit (7.0g) of described process scale material, efflorescence, and this material water (250mL) extracted 4 hours in 25 ℃.Mixture is filtered, and filter liquor is concentrated into drying, obtain the room temperature water extract (4.2g) of exsiccant fruit by rotary evaporation.

FD001 (79.9g) is mixed with the 1.5L distilled water, and this solution is extracted four times in turn with each 500mL ethyl acetate (EtOAc).The organic layer and the water layer that merge are concentrated into drying in a vacuum, obtain EtOAc extract (7.4g) and aqueous residue (41.5g).

Embodiment 4

The following examples have been described the active testing in vitro of immunoregulation of Actinidia arguta Sieb.et Zucc prepared product.

The purpose of this research is: by ELISA (Quantikine kits, R﹠amp; D systems) analyzes, relatively various extracts that make from Actinidia arguta Sieb.et Zucc and prepared product are from ovalbumin (OVA, V level, Sigma) relative ability of regulating cell factor generation (IL-4, IL-5, IL-10, IL-13 and IFN γ) in the spleen cell cultures thing of sensitization mice.Tested following sample (preparation described in the embodiment 3 as mentioned): FD001 (PG102T), fruit juice concentrates and EtOAc extract.

The separation of splenocyte and cultivation

(Harlan, Indianapolis is IN) by carrying out sensitization at the 0th and 14 day peritoneal injection 20 μ g OVA with female Balb/c mice.At the 24th day, implement euthanasia by cervical dislocation after, under aseptic condition, take out spleen and exist side by side and promptly use aseptic technique processing for spleen cell cultures from each mice.Under the buffered RPMI-1640 existence condition of 10mMHEPES, force tissue spleen to be dissociated gently by the grid of 70 micrometer nylon screen clothes with the plunger of 3cc syringe.Use the FCS gradient from the gained suspension, to remove bigger cell aggregation thing.Centrifugal (1500rpm 5min), and handles gained cell precipitation thing that (10min is RT) to remove the erythrocyte of sneaking into the RBC lysis buffer with splenocyte then.(1500rpm 5min) removes most of RBC lysis buffer, then sedimentary splenocyte is cleaned 3 times with the buffered RPMI-1640 of 10mM HEPES again by centrifugal then.After last the cleaning, sedimentary splenocyte is resuspended in the RPMI-1640 that contains 10%FCS and penicillin/streptomycin (complete medium) of certain volume, to produce 5 * 10 ⁶The final cell density of individual cell/mL.Analyze 5 * 10 at every turn ⁶Individual splenocyte is assigned in each hole of 24 orifice plates.At the 3rd day, the supernatant of collecting these holes was also freezing, prepares to be used for the determination experiment result.

Also set up contrast spleen cell cultures thing from (not sensitization) mice of naivety in the manner described above, and be assigned in each hole of 24 orifice plates, reached 5 * 10 ⁶The final cell density of individual cell/mL.These splenocytes are to set up in the RPMI-1640 that contains 10% hyclone, penicillin/streptomycin, do not accept other processing.At the 3rd day, the supernatant of collecting these holes was also freezing, serves as negative experiment contrast.

Stimulate the spleen cell cultures thing with the Actinidia arguta Sieb.et Zucc prepared product

Every kind of prepared product is tested with the mice of 10 OVA sensitizations.To be assigned in 8 holes of 24 orifice plates (5 * 10 from the splenocyte of every mice ⁶Individual cell/mL), wherein cell is in and contains 100 μ g/mLOVA, 0.5%DMSO and not or have in the complete RPMI-1640 culture medium of fc-specific test FC prepared product of every kind of check of selected concentration.Be divided into 2 groups, each 3 hole with 6 in 8 holes.With these each holes of 3 one group with 0.25,1.0 or the Actinidia arguta Sieb.et Zucc prepared product of 10mg/mL concentration handle.The 7th hole is with 2 μ M dexamethasone (DEX)--a kind of strong glucocorticoid antiinflammatory is handled as positive control.The 8th hole do not accepted other and handled, and serves as the experiment contrast that has only OVA.After cultivating 3 days, that the supernatant of each in every distinctive 8 holes of OVA sensitization mice of collection is also freezing.These supernatant are used for measuring the level that cytokine IL-4, IL-5, IL-10, IL-13 and IFN-γ exist at the culture culture fluid.

The mensuration of cytokine levels in the culture supernatants

Measure the splenocyte handled derived from the Actinidia arguta Sieb.et Zucc prepared product, splenocyte that DEX handles, the splenocyte of only handling with OVA and from the cytokine levels in the culture supernatants of the splenocyte that is untreated of not sensitization control mice by the ELISA algoscopy.The mensuration of every kind of cytokine levels is used the hole of duplicate elisa plate.

The result

This external work has confirmed the activity in the FD001 (Figure 11) suitable with PG102T, EtOAc extract (Figure 12) and the fruit juice concentrates (Figure 13) of Actinidia arguta Sieb.et Zucc.Observing all three kinds of prepared products (at 10mg/mL) all causes the substantive in various degree of pair cell factor IL-4, IL-5, IL-10, IL-13 and IFN γ to suppress.For all samples of check, on IL-13 and IFN-γ, see the most significant effect, with the previous consistent (table 1 of external work; Embodiment 1).Owing to observe activity in the EtOAc extract, obviously the active component that exists among the FD001 can extract in the organic solvent, and can be further purified by traditional chromatography.Importantly, fruit juice concentrates also suppresses the splenocyte cellulation factor, shows that extracting kiwi fruit as shown in embodiment 1 and 2 is not the unique requirement that generates active hard kiwi fruit prepared product.The inhibition of noticing the fruit juice concentrates pair cell factor is obviously relatively low, illustrates that the drying or the heating process that are used to prepare FD001 may be important for improving activity.

Embodiment 5

The following examples have been described the non-fruit extracting section of Actinidia arguta Sieb.et Zucc thing, and the comparison of the external activity of other Actinidia arguta Sieb.et Zucc fruit prepared product.

The purpose of this research is: utilize elisa assay, assessment derives from the extract of actinidia arguta of the plant part beyond the fruit or the ability that other fruit prepared product (promptly different with extract described in embodiment 1 and 2) regulating cell factor in derived from the spleen cell cultures thing of ovalbumin sensitization mice generates (IL-13 and IFN γ).Tested following sample (preparation as indicated above): the water extract of the stem of Actinidia arguta Sieb.et Zucc, root, bark, preparation as described in example 3 above; " boiled " fresh fruit prepared product; Fruit juice concentrates, preparation as described in example 3 above; FD001 (the extensive quite thing of PG102T), preparation as described in example 3 above; FD001 powder, preparation (being used for clinical trial described below) as described in example 3 above; The room temperature water extract of exsiccant Actinidia arguta Sieb.et Zucc fruit; EtOAc extract, preparation as described in example 3 above; And water surplus, description is also arranged in embodiment 3.In addition, assess three kinds of known inhibitive ability of immunity chemical compounds, promptly the activity of cyclosporin, dexamethasone and Quercetin (quercetin) in contrast.

The separation of splenocyte and cultivation

The preparation of splenocyte is carried out in the mode identical with mode described in the embodiment 4 above.

Stimulate the spleen cell cultures thing with extract of actinidia arguta

Every kind of extract of test or prepared product use and analyze from the splenocyte of 8 OVA sensitization mices (being 8 parts of the same form).With 5 * 10 ⁶Individual splenocyte from every mice is assigned in each hole of 24 orifice plates, and wherein cell is in the complete RPMI-1640 culture medium that contains 100 μ g/mL OVA and 25mM HEPES (pH 7.3), every hole 1ml.The check concentration of kiwi fruit prepared product is 1.0,3.0 and 10mg/mL.

Cyclosporin, Quercetin and dexamethasone analysis

Every kind of chemical compound of test uses and analyzes from the splenocyte of 8 OVA sensitization mices (being 8 parts of the same form), and exception is that Quercetin is only used from the splenocyte of 2 OVA sensitization mices and tested.With 5 * 10 ⁶Individual splenocyte from each mice is assigned in each hole of 24 orifice plates, and wherein cell is in the complete RPMI-1640 culture medium that contains 100 μ g/mL OVA and 25mM HEPES (pH 7.3), every hole 1ml.The check concentration of cyclosporin is 0.0083,0.083 and 4.15 μ M.The check concentration of dexamethasone is 0.01,0.1 and 1 μ M.The check concentration of Quercetin is 1,10 and 25 μ M.The hole of positive experiment contrast for handling with 1 μ M dexamethasone (a kind of strong glucocorticoid antiinflammatory).Only accept to contain the hole of the complete RPMI-1640 of 100 μ g/ml OVA and 25mM HEPES (pH 7.3) and serve as the only experiment contrast of usefulness OVA.Cultivate and collect supernatant and freezing after 3 days.Utilize these supernatant mensuration IL-13 and IFN-γ in the level that in various culture culture fluid, exists under the experiment condition of being checked.

The mensuration of cytokine levels in the culture supernatants

Measure from the cytokine levels in the culture supernatants of all processing holes and control wells by the ELISA algoscopy.The mensuration of every kind of cytokine levels is used the hole of duplicate elisa plate.

According to the result of the testing in vitro described in the embodiment 4,, only analyzed the expression of IL-13 and IFN-γ for the purpose of estimating the activity level that exists in the material of testing.

The result

Prove that as previous the concentration of observing with the Actinidia arguta Sieb.et Zucc test substances raises, to the inhibition increase of check cytokine.Generally speaking, more remarkable at the inhibition of IFN γ.In this algoscopy, the activity of prescription inhibitive ability of immunity chemical compound is similar to the Actinidia arguta Sieb.et Zucc prepared product.Peptide cyclosporin and glucocorticoid steroid dexamethasone show strong activity (＜1 μ M), shown in Figure 14 A.The flavonoid Quercetin demonstrates strong activity (1-25 μ M, Figure 14 B) in higher slightly concentration range.Shown EtOAc extracts active ability from FD001 further confirmation among Figure 15 A and the 15B.What is interesting is, also observe existence activity in the aqueous residue, show that polarity and nonpolarity element all may be the reasons of external immunosuppressive action.Shown in Figure 16 A and 16B, also proved the FD001 powder this in dog and people's clinical trial (hereinafter described) all used material in this mensuration, be activated.

As described in example 4 above, explored other different and prepared the method for kiwi fruit extract with flow process described in embodiment 1 and 2.No matter the deutero-extract of all fruits is exsiccant or fresh, no matter extracts in hot water or room temperature water, all shows similar activity in this algoscopy, shown in Figure 17 A and 17B.According to this analysis, the inventor thinks has several reliable alternative methods to can be used for preparing the kiwi fruit that uses for therapeutic purposes.

What is interesting is, this algoscopy, show with FD001 (PG102T) from the hot water extract of the bark of Actinidia arguta Sieb.et Zucc, root and stem preparation and compare equal or better activity (Figure 18 A and 18B) with fruit juice concentrates.These results show, at regulation and control immunological marker thing or suppress aspect the pro-inflammatory cytokine, these other plant part may become the alternative source of the chemical compound that the treatment benefit is arranged.

Embodiment 6

The following examples have described the double blinding carried out in medium order of severity atopic dermatitis adult experimenter, be the result that the FD001 (PG102T) of contrast renders a service out-patient's research with the placebo.

The purpose of this research is to obtain FD001 (PG102T) in the primary evidence of suffering from the effectiveness among the adult volunteer of medium order of severity atopic dermatitis (AD) after Orally administered 42 days in minority.Second purpose of this research is tolerability and the response transmutability of assessment to FD001.

Research design

The experimenter takes FD001 powder (preparation as described in example 3 above), and 2 600mg capsules in the morning (FD001 accumulated dose 600mg) or two Cebo-Capses (being made up of MCC) continue 42 days, certainly beginning in the 1st day of research.The indication experimenter stopped using the steroid medication after the 14th day.Gather blood at 4 time points and be used for routine biochemistry group and analysis of Hematology Changes: when visiting and the 1st, 14 and 42 day of research for the screening experimenter.Measured the level of IgE and c reactive protein matter in the blood at the 1st, 14 and 42 day.These 4 days collection urines are used for routine urinalysis at all.Carry out efficacy assessment during each visit after screening.Experimenter or male or women, 19 to 65 years old, health status was generally good.The experimenter suffers from according to the doctor physician overall evaluation (Feldman and Krueger, Annals of theRheumatic Diseases, 64:ii65-ii68,2005) in 0 to 4 order of severity grade, must be divided into the activeness atopic dermatitis of the medium orders of severity of 3 definition.The experimenter suffers from the minimum AD that relates to 10% body surface area (BSA).The experimenter is using topical steroids with treatment AD at present, and can not be just in suckling or pregnancy.Use untoward reaction report and standard blood chemistry, hematology and urinalysis to assess safety and tolerability.

Statistical method

Primary efficacy variable be in the time of 42 days the doctor physician overall evaluation with respect to the variation of baseline, use Cochran Mantel-Haenszel check to analyze (Armitage et al., Statistical Methods inMedical Research, 4th Ed., Blackwell, Oxford, 2002).Secondary variable be AD sign in the time of 42 days (erythema, harden, ooze out/form a scab and the pruritus severity scale) and total BSA with respect to the variation of baseline, use two sample t check to analyze.Provide treatment in the 1st, 14,28 and 42 day to organize the descriptive statistic data of data behind all baselines and the baseline.As required, these statistical items comprise sample size, meansigma methods, standard deviation, frequency, percentage rate and confidence interval.By treatment group statistics with result from experimenter's self evaluation application form is provided.Any adverse events that takes place during the record research.The description of adverse events comprises the order of severity, attribution, the action of taking, the therapy of taking, and the result of date of breaking-out day, incident finishing or continue, incident.With experimenter's number of these data based report adverse events, airframe systems (body system), the order of severity, seriousness, and classify with the relation of tester.MedDRA project (Medical Dictionary forRegulatory Activities will have taken place to be included into during will studying, the experimenter's of one or more adverse events http://www.meddramsso.com) list of frequency compares between each treatment group.

All determined the descriptive summary statistical data of laboratory values and with respect to the associated change of baseline (the 1st day) for the assessment of all clinical laboratories.In data list labelling the numerical value beyond the normal range.In addition, make " change list " and be presented at the experimenter's that laboratory parameters changes during the research number and percentage ratio (for example based on laboratory with reference to scope, by normally becoming height).

Table 5

^*All experimenters of A=, B=remove clobetasol (clobestasol) and clobetasol propionate (ultravate) user, and C=has only the user of HDRCRT-17-hydroxy-11-dehydrocorticosterone (cortoid) or triamcinolone (triamcinolone).

Efficacy outcomes

17 experimenters of pro-carry out interim analysis to finish research, and the result draws intensive statistics tendency in two secondary efficacy terminal points: erythema (p=0.13, ITT 17) and sclerosis (p=0.09, ITT 17).When interim analysis, detect in so little sample size less than the significant difference in the main effect terminal point.

During last analysis, in primary efficacy variable (the doctor physician overall evaluation) or secondary efficacy variable (AD sign with respect to the variation of baseline or the percentage ratio of BSA change) there be not performance statistics significance between each treatment group at the 42nd day.Yet,, in main effect terminal point, observe significant changes in distribution at the 14th day.Placebo and tester receiver's this variation that takes place of replying between the 1st day and the 14th day is found in respectively among Figure 19 A and the 19B and (has indicated the PGA standards of grading).When with regard to the 14th day analysis result the experimenter being included into the response group and not responding group (table 5, A group), as seen more a high proportion of respondent in the FD001 processed group, and not the respondent to account for the ratio of placebo group higher.Take in eliminating (B group) behind the experimenter of more potent steroid, and for only accepting the experimenter (C group) of moderate to slight steroid therapy, these results keep unanimity relatively.The significance,statistical (p=0.02) of main PGA terminal point and experimenter were to the significance,statistical of rubescent self evaluation (p=0.03) when the cause and effect of complementarity (posthoc) analysis had been disclosed the 14th day.When in addition, detecting the 14th day all there are intensive statistics tendency (being respectively p=0.08 and p=0.07) in the self evaluation of itching, the clinical sign assessment that reaches oozing out/forming a scab.The 14th day by the PGA endpoint observation to the effect to the compared with placebo group of processed group disappeared in 42 days.These results show the treatment of using FD001 (PG102T) can help AD as the complementary therapy of topical corticosteroid therapy.1st, 14 show any test of being carried out tendency relevant with safety not with 42 days laboratory test results, comprise clinical chemistry, complete blood count and segmentation, blood coagulation, unconjugated bilirubin and urine macroscopic view.IgE, c reactive protein matter or eosinophil count do not find differences between each processed group.

Safety results

All do not report serious adverse events to FD001 processing or placebo.FD001 has reported 12 examples not serious incident, and placebo is 13 examples.Wherein, for FD001, to moderate, 2 examples are the severe incident to 10 examples for slightly; Placebo is that 13 examples are slightly to the moderate incident.Think that it is possible or relevant with research tester or placebo not having which routine incident.

Conclusion

The analysis result of plan proved in the time of the 42nd day does not have significance,statistical between the treatment group in main terminal point and secondary endpoints.Yet, in main terminal point (PGA), noticed intensive changes in distribution (distributional shift) at the 14th day.For this reason, this time point is carried out cause and effect (post hoc) analysis, had the significant difference of statistics between the processed group when this analysis is disclosed in the 14th day time point.In addition, some secondary endpoints shows statistically significant (erythema p=0.03, self evaluation) or intensive statistically tendency (is itched and improved p=0.08, self evaluation; And ooze out/form a scab p=0.07, the AD sign is estimated).Do not report in test serious adverse events.The adverse events number of reporting in tester group and the placebo group is suitable, and can not be owing to tester.In clinical laboratory's numerical value, do not find safety issue.These data have been affirmed clinical preceding small-sized mammalian and the suggested conclusion of unmatchful photograph (anecdotal) human research once more, and promptly the Actinidia arguta Sieb.et Zucc fruit extract is safe to the receiver, and is well tolerated.

Embodiment 7

The following examples have been described a hard kiwi fruit extract of assessment and have been used to reduce atopy dog CADESI score (Olivry et al., Vet.Dermatol., 13:77-87,2002; Hanifm et al., Exp.Dermatol., 10:11-18,2001; Kunz et al., Dermatology, 195:10-19,1997) randomization, double blinding, placebo-controlled study.

The purpose of this research is the effect of assessment Actinidia arguta Sieb.et Zucc fruit extract FD001 (PG102T) as the complementary therapy of atopic dermatitis in the dog (AD) standard steroid therapy.Use investigator's the overall evaluation to assess response, wherein comprised the CADESI grade and the owner (owner ' s) pruritus evaluation treatment.Another purpose of this research is that assessment kiwi fruit extract reduces the effect of steroid demand in the management of AD clinical sign as monotherapy, and assesses safety.

Research design

Give only to have in two weeks low dosage (0.2mg/kg) steroid (prednisolone) during, to measure the response of steroid in dog, keep all the other AD symptoms simultaneously.All dogs all are healthy, get rid of non-seasonal AD dermatosis.The diagnosis of AD comprises at least three positive reactions to the test of Intradermal skin allergy.Dog is accepted flea control treatment (Advantage), and to stand diet test be the main cause of symptomatology to get rid of food anaphylaxis.Dog is not accepted medication together, such as hydryllin and long-acting injection steroid, and does not have secondary infection.Must be at least 25 by baseline (the 1st day) score of CADESI grade and can include research range in.

During auxiliary treatment (14-42 days), dog is the also oral every other day prednisolone (0.2mg/kg) of accepting except tester.Research experimenter (it is 60 that target is recruited number) consumes FD001 powder (preparation as described in example 4 above) or placebo (MCC) with 30mg/kg dosage once a day, continues 28 days.The experimenter is randomized into tester group and placebo group with 1: 1 ratio.Using CADESI grade and the grading of owner's pruritus that the experimenter is assessed on the record weekly.The the 1st and 28 day collecting blood sample of during only steroid being arranged the 1st day and research is with assessment safety and secondary endpoints.The analysis project of these samples comprises measures complete blood count, clinical chemistry, total IgE, allergen specific IgE and chemotactic factor TARC.

The complementary therapy treatment phase for the dog that any performance overall evaluation improves, is proceeded 28 days open-label (open label) second stage research when finishing, and is made up of as the monotherapy of AD FD001 tester (30mg/kg/ day).The recurrence of AD sign taking place or need the experimenter of retrospective medication will be considered as second stage treatment loser, and withdraws from research.The experimenter who keeps its improvement in second stage will be considered as treating the respondent.Carried out in per 7 days the CADESI grade and every day pruritus record assessment.The the 14th and 28 day collecting blood sample in second stage.Identical with phase I research analysed in laboratory blood constitutent.

Statistical method

Main efficiency analysis is according to investigator's the overall evaluation treatment to be had the experimenter's of positive response ratio, wherein uses X 2 test.When finishing research, half that can recruit number in target carry out interim analysis.P＜0.05 is considered as significantly.

Conclusion

Atopic dermatitis is the second common allergy in the dog, and generation (Scott et al., Small Animal Dermatology, 5th Ed., WB Saunders, 500-518,1995) is arranged in about 10% the individuality in the dog group.Be typically, the AD in the dog trends towards worsening with the age increase.Infected animal suffers skin and ear infection repeatedly, greatly reduces their quality of life.Although AD is very common disease, yet available treatment selection is limited.Systematicness is handled, and as glucocorticoid and cyclosporin, may be effectively, but may have untoward reaction (Olivry et al., Vet.Dermatol, 13:77-87,2002; Rvffel, et al., Arch.Toxicol., 53:107-141,1983).Glucocorticoid trends towards rendeing a service reduction with life-time service, oral cyclosporin is the possibility both expensive in the dog of large-scale kind, and usually lower (the Scott et al. of antihistaminic success rate, Small Animal Dermatology, 5th Ed., WBSaunders, 500-518,1995).Identify that safe and efficient Therapeutic Method will be extremely beneficial with the S﹠S that alleviates dog AD.FD001 can reduce in the dog AD management to the demand of steroid as auxiliary treatment or as the effect of monotherapy.Also having looked forward to the kiwi fruit extract will be as complementary therapy to reduce the effectiveness of used steroid.Can imagine described extract can be used as capsule, with blended powder of food or food become to assign to use.When replenishing independent use or associating low dosage steroid, the diet that contains the kiwi fruit prepared product can in having atopic dog, effectively support the skin of health.

Embodiment 8

The following examples have been described the purposes that Actinidia arguta Sieb.et Zucc and extract thereof are used for regulating and control the immunne response of allergia and nonallergic inflammatory disease.

The purpose of this research is to obtain FD001 (PG102T) to suffer from the primary evidence of medium order of severity allergic disease such as the effectiveness among the adult volunteer of atopic dermatitis (AD), asthma or allergic rhinitis after Orally administered 28 days.Second purpose of this research is to measure the bioavailability of this product when use substitutes delivery form (for example capsule, the concentrate that is added into beverage, Emulsion or food component).Another object of the present invention is to measure FD001 to the level of short inflammatory blood mark such as cytokine, chemotactic factor, leukotriene or antibody, to the propagation of bone marrow like cell (for example leukocyte, macrophage, mastocyte etc.), and to the influence of related gene expression and transcription factor.

Carry out the out-patient research of double blinding, placebo, wherein the experimenter is used FD001 or placebo as tester.The treatment group of this research comprises that FD001 is as monotherapy or as the oral slight extremely complementary therapy of the steroid of middle equivalent force.Perhaps, AD experimenter's steroid therapy can be partial.For asthma and allergic rhinitis experimenter, the use of steroid can be respectively as the form of inhaler or intranasal spraying.The experimenter includes standard, efficacy assessment, safety, tolerability and statistical analysis use in and assesses with the similar method of method described in the embodiment 6.

Allergic a kind of performance--atopic dermatitis is a common skin disease among the child, is found in be born 6 middle of the month (Spergel and Paller, J.Allergy Clin.Immunol., 112:S128-S139,2003) usually.As if the popularity degree of AD increases day by day in the whole world, and other atopy disease comprises that asthma also is (Larsen and Hanikin, Immunology and Allergy Clinicsof North America, 22:1-25,2002 like this; Wollenberg et al., Clin.Exp.Dermatol, 25:530-534,2000; Mannino et al., Mor Mortal Wkly Rep CDC Surveill Summ., 47:1-27,1998; Linneberg et al., Allergy, 55:767-772,2000).AD patient's quality of life suffers serious harmful effect, and existing at present Therapeutic Method both may be the source of adverse side effect, was again the financial burden of family and society.Atopic cutaneous manifestations has usually been represented the beginning of atopy development.Based on several tracing properties (longitudinal) research, approximately asthma will take place in half AD patient, severe AD particularly, and 2/3rds allergic rhinitis (Leung et al., J.Clin.Invest., 113:651-657,2004 will take place; Spergel et al., Clin.Invest., 101:1614-1622,1998).The discovery of AD Therapeutic Method safely and effectively will be subjected to great welcome.The effect of FD001 also can reduce steroid or used amount or the effectiveness of other medicines in AD, asthma or allergic rhinitis management.

As the exercise question of later test, the extract that can also look forward to kiwi fruit extract, concentrate, other prepared product or other plant part (for example bark, stem, root, leaf) is used for the purposes of AD, asthma, allergic rhinitis or other leukotriene mediation illness such as food anaphylaxis and chronic urticaria as auxiliary treatment or monotherapy.As the reasonable extension of this work, can further explore the purposes of kiwi fruit product as the Therapeutic Method of allergia illness in the small-sized mammalian (dog for example, also can referring to embodiment 7).

At this with each piece list of references described herein or the complete this paper that incorporates into of publication.

Although described a plurality of embodiment of the present invention in detail, the modification of those embodiments and adaptive will be conspicuous to those skilled in the art.Yet, the understanding that should be understood that, this type of modification and fitting within the scope of the invention listed in the appended exemplary claim.

Sequence table

＜110〉Efficas Inc (EFFICAS INC.)

Lindemann，Julianne

Stagnitti，George?E.

Driver，Robert?H.

Braman，Mark?A.

Fogg-Johnson，Nancy?E.

Kim，Sunyoung

Park，Eun-Jin

Kim，Bongcheol

Jin，Mirim

Jung，Hyung-Jin

Shin，Sung-Seup

Oh，Jin-Hwan

Lee，Hwa-Jun

Jeon，Hyang

＜120〉comprise the compoistion and method of use of actinidia

<130>5224-9-PCT

＜140〉herewith submit to

<141>2006-02-24

<150>60/656,839

<151>2005-02-25

<150>60/656,838

<151>2005-02-25

<160>8

<170>PatentIn?version?3.3

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Claims (67)

1. regulate and control the method for the immunne response in the mammal, comprise with the amount that is enough to regulate and control the immunne response in the mammal the hard kiwi fruit prepared product of administration, wherein said hard kiwi fruit prepared product is selected from: fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, spissated fruit, the dry fruit of crossing and hard kiwi fruit fruit juice concentrates, wherein said hard kiwi fruit prepared product un-extracted.

2. regulate and control the method for the immunne response in the mammal, comprise that with the amount that is enough to regulate and control the immunne response in the mammal to the hard kiwi fruit prepared product of administration, wherein said hard kiwi fruit prepared product is selected from: leaf extract or concentrate, and bark extract or concentrate.

3. the method for the immunne response of regulation and control in the mammal comprises with the amount that is enough to regulate and control the immunne response in this mammal administration: a) hard kiwi fruit prepared product; And b) is selected from down the composition of organizing: probiotics; Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; And proanthocyanidin.

4. the method for the immunne response of regulation and control in the mammal comprises with the amount that is enough to regulate and control the immunne response in this mammal administration: a) hard kiwi fruit prepared product; And b) is selected from down the composition of group: steroid, hydryllin, antibody, antibiotic, cyclosporin, antifungal, respiratory function regulation agent, analgesic, beta-2-agonists, leukotrienes regulator, cytokine or cytokine receptor antagonist, phosphodiesterase inhibitor, sodium cromoglicate, nedocromil, caffeine, theophylline, benzyloxycarbonyl group β-alanyltaurine, reach T cell function inhibitor.

5. claim 3 or 4 method, wherein said hard kiwi fruit prepared product is selected from: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, spissated fruit, the dry fruit of crossing, hard kiwi fruit fruit juice concentrates, in about 80 ℃ water, extract the prepared product that fruit obtains by in temperature being 0 ℃, by directly extracting the prepared product that hard kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that hard kiwi fruit obtains, and pass through at water, extract the prepared product that hard kiwi fruit obtains in chloroform and the ethyl acetate in turn.

6. each method of claim 3-5, wherein said hard kiwi fruit prepared product is extract or the concentrate that partly prepares from the hard kiwi fruit that is selected from down group: leaf, stem, bark, root and combination in any thereof.

7. each method of claim 1 or 3-5, wherein said hard kiwi fruit prepared product is selected from: fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, reach spissated fruit.

8. each method of claim 1 or 3-5, wherein said hard kiwi fruit is the dry fruit of crossing.

9. each method of claim 1 or 3-5, wherein said hard kiwi fruit prepared product are that the method by the step that comprises dry fruit obtains.

10. each method of claim 1 or 3-5, wherein said hard kiwi fruit prepared product is hard kiwi fruit fruit juice concentrates.

11. each method of claim 3-5, wherein said hard kiwi fruit prepared product are to extract fruit in about 25 ℃ water and obtain by being about 0 ℃ in temperature.

12. each method of claim 3-5, wherein said hard kiwi fruit prepared product are to obtain by extract fruit in the water of room temperature.

13. each method of claim 3-5, wherein said hard kiwi fruit prepared product obtains by directly extracting hard kiwi fruit water-soluble concentrate with ethyl acetate.

14. each method of claim 1-13, the amount of the kiwi fruit prepared product that is wherein provided is enough to regulate and control Th2 and the Th1 immunne response in the mammal.

15. each method of claim 1-13, the amount of the kiwi fruit prepared product that is wherein provided is enough to regulate and control that mammal generates, as to be selected from the antibody isotype of IgE, IgG2a and IgG1 amount.

16. each method of claim 1-13, the amount of the kiwi fruit prepared product that is wherein provided are enough to reduce the generation and/or the level of at least a Th2 cytokine in the mammal or improve the level of at least a Th1 cytokine in the mammal.

17. each method of claim 1-13, the amount of the kiwi fruit prepared product that is wherein provided is enough to reduce the level or the generation of at least a leukotriene in the mammal.

18. each method of claim 1-13, the amount of the kiwi fruit prepared product that is wherein provided is enough to reduce the expression of the transcription factor that is selected from GATA-3, T-bet and NFATc2 in the mammal.

19. each method of claim 1-18, wherein said mammal suffers from or risky generation to strengthen wherein that Th1 replys and/or suppress that Th2 replys be favourable illness.

20. the method for claim 19, wherein said mammal suffer from or risky generation allergia or nonallergic inflammatory disease.

21. the method for claim 20, wherein said allergic disease is regulated and control by leukotriene.

22. the method for claim 20, wherein said allergic disease is an asthma.

23. the method for claim 20, wherein said allergic disease is an atopic dermatitis.

24. the method for claim 19, wherein said mammal suffer from or risky generation viral infection.

25. the method for claim 19, wherein said mammal suffer from or risky generation cancer.

26. each method of claim 1-25, wherein said hard kiwi fruit is selected from Actinidia arguta Sieb.et Zucc, ovateleaf actinidia leaf and Semen Actinidiae Polygamae.

27. each method of claim 1-25, wherein said hard kiwi fruit is an Actinidia arguta Sieb.et Zucc.

28. each method of claim 1-27, wherein said hard kiwi fruit prepared product provides in compositions, and its amount accounts for about 0.01% to about 95% of described composition total weight by weight.

29. each method of claim 1-28, wherein step of applying comprises with carrier, adjuvant or diluent the hard kiwi fruit prepared product of administration.

30. each method of claim 1-29, wherein step of applying comprises hard kiwi fruit prepared product is offered mammal as tablet, powder, effervescent tablet, effervescent powder, capsule, liquid, suspension, granule or syrup.

31. each method of claim 1-29, wherein step of applying is included in the health food and provides hard kiwi fruit prepared product to mammal.

32. the method for claim 31, wherein said health food is selected from: bakery product, bread, volume stuffs, breakfast cereals, finished cheese, the cheese of undressed mistake, flavoring agent, milk product, pudding, gelatin jelly, soda pop, the teas beverage, drink dried powder, finished fish product, beverage based on fruit, beverage based on vegetable, chewing gum, hard sugar, refrigerated milk product, finished meat products, coating based on nut, wheaten food, finished poultry product, thick gravy and sauce, potato chips, vegetable chips, crispy slice, chocolate, cookies, confection, Liquoride sugar, ice cream, dehydrated food, the food of cutting, finished food, spice, alcoholic beverage, noodles, fermented food, soup, powder, bean product, coating based on vegetable oil, with beverage based on vegetable.

33. each method of claim 1-29, wherein step of applying comprises the cosmetic composition that administration is comprised hard kiwi fruit prepared product.

34. the method for claim 33, wherein said cosmetic composition are to provide with the form that is selected from down group: beauty treatment emulsion, cream, essence, skin contraction agent, Emulsion, beauty treatment Liniment, soap, shampoo, irrigation, cleaning agent, bath foam, washing liquid or inorganic agent.

35. each method of claim 1-29, wherein step of applying is included in the food additive and provides hard kiwi fruit prepared product to mammal.

36. each method of claim 1-35 further comprises the medicine that administration is selected from down group: fatty acid; Polyketide; Organic acid; Little organic compound; Aromatic amino acid; The phenylpropyl alcohol alkyl compound; Terpenoid; Steroid; Alkaloid; Corrin; Porphyrin; The line peptide; Cyclic peptide; The ester peptide; Amino acid derivativges; Nucleoside; Nucleotide; Carbohydrate; Protein; Cell; Cell debris; The medical herbs prepared product; Spice; Mineral; Antibacterial; Flavoring agent; Vitamin; And electrolyte.

37. each method of claim 1-35 further comprises the medicine that administration is selected from down group: probiotics; Bacteria cell wall and fragment; Whey protein; Taurine; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Herba Rosmarini Officinalis; Rosmarinic acid; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; Proanthocyanidin; And beta-carotene.

38. the method for claim 37, wherein said fatty acid is selected from: conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid.

39. each method of claim 1-38 further comprises the prepared product to other Actinidia species of administration.

40. the method for claim 39, wherein said other Actinidia species are selected from: A.chinensis Planch., delicious Fructus actinidiae chinensis, Actinidia arguta Sieb.et Zucc, Semen Actinidiae Polygamae and ovateleaf actinidia leaf.

41. be used for regulating and control the compositions of mammal immunne response, comprise hard kiwi fruit prepared product and at least a other reactive compound that is used for regulating and control the mammal immunne response.

42. the compositions of claim 41, wherein said other reactive compound are used for the treatment of or prevent allergic disease in the mammal.

43. the compositions of claim 41, wherein said other reactive compound is selected from: steroid, hydryllin, antibody, antibiotic, cyclosporin, antifungal, respiratory function regulation agent, analgesic, beta-2-agonists, leukotrienes regulator, cytokine or cytokine receptor antagonist, phosphodiesterase inhibitor, sodium cromoglicate, nedocromil, caffeine, theophylline, benzyloxycarbonyl group β-alanyltaurine and T cell function inhibitor.

44. the compositions of claim 41, wherein said other reactive compound is selected from: probiotics; Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; And proanthocyanidin.

45. the compositions of claim 44, wherein said fatty acid is selected from: conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid.

46. the compositions of claim 41, wherein said compositions is a Pharmaceutical composition.

47. the compositions of claim 40, wherein said compositions is a health food.

48. the compositions of claim 41, wherein said compositions is a food additive.

49. the compositions of claim 41, wherein said compositions is cosmetics.

50. each compositions of claim 41-49, wherein said hard kiwi fruit is selected from Actinidia arguta Sieb.et Zucc, ovateleaf actinidia leaf and Semen Actinidiae Polygamae.

51. each compositions of claim 41-49, wherein said hard kiwi fruit is an Actinidia arguta Sieb.et Zucc.

52. each compositions of claim 41-49, wherein said hard kiwi fruit prepared product are extract or the concentrate that partly prepares from the hard kiwi fruit that is selected from down group: fruit, leaf, stem, bark, root and combination in any thereof.

53. each compositions of claim 41-49, wherein said hard kiwi fruit is selected from: fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing and spissated fruit.

54. each compositions of claim 41-49, wherein said hard kiwi fruit are the dry fruits of crossing.

55. being the methods by the step that comprises dry fruit, each compositions of claim 41-49, wherein said hard kiwi fruit prepared product obtain.

56. each compositions of claim 41-49, wherein said hard kiwi fruit prepared product is hard kiwi fruit fruit juice concentrates.

57. each compositions of claim 41-49, wherein said hard kiwi fruit prepared product are to obtain by extract fruit in the water of room temperature.

58. each compositions of claim 41-49, wherein said hard kiwi fruit prepared product obtains by directly extracting hard kiwi fruit water-soluble concentrate with ethyl acetate.

59. each compositions of claim 41-49, wherein said extract obtains by extract hard kiwi fruit in distilled water.

60. each compositions of claim 41-49, wherein said extract is the ethyl acetate extract of hard kiwi fruit.

61. the medicine that hard kiwi fruit or its prepared product and being selected from down organized is used for the treatment of purposes in the compositions with immune dysfunction diseases associated or illness in preparation: steroid; hydryllin; antibody; antibiotic; cyclosporin; antifungal; respiratory function regulation agent; analgesic; beta-2-agonists; leukotrienes regulator; cytokine antagonist; cytokine receptor antagonist; phosphodiesterase inhibitor; sodium cromoglicate; nedocromil; caffeine; theophylline; benzyloxycarbonyl group β-alanyltaurine; with T cell function inhibitor.

62. hard kiwi fruit or its prepared product and be selected from down the purposes of the medicine of organizing: probiotics; Bacteria cell wall and fragment; Whey protein; Alanine; Fatty acid; Fatty acid ester; Monoglyceride; Diglyceride; Triglyceride; Inositol; Rhizoma Curcumae Longae; Curcumin; Dimethylsulfone (MSM); Radix Ginseng; Rhizoma Zingiberis Recens; And proanthocyanidin.

63. the purposes of claim 62, wherein said fatty acid is selected from: conjugated linolenic acid, eicosapentaenoic acid, docosahexenoic acid, gamma-Linolenic acid, alpha-linolenic acid, bishomo-and parinaric acid.

64. each purposes of claim 61-63, wherein said compositions are used for the treatment of and leukotriene generates or active diseases associated or illness.

65. each purposes of claim 61-63, wherein said disease or illness are selected from: atopic dermatitis, asthma, food anaphylaxis, allergic rhinitis and chronic urticaria.

66. each purposes of claim 61-63, wherein said hard kiwi fruit prepared product is selected from: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, spissated fruit, the dry fruit of crossing, hard kiwi fruit fruit juice concentrates, by in the water of room temperature, extracting the prepared product that fruit obtains, by directly extracting the prepared product that hard kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that hard kiwi fruit obtains, with by at water, extract the prepared product that obtains in chloroform and the ethyl acetate in turn.

67. the method for immunne response in the regulation and control mammal, comprise that with the amount that is enough to regulate and control immunne response in the mammal to the common kiwi fruit prepared product of administration, wherein said common kiwi fruit prepared product is selected from: fruit extract or concentrate, leaf extract or concentrate, stem extract or concentrate, bark extract or concentrate, root extract or concentrate, fresh fruit, broken fruit, the fruit that boiled, the fruit of cooking, the fruit of pressing, spissated fruit, the dry fruit of crossing, common kiwi fruit fruit juice concentrates, by in the water of room temperature, extracting the prepared product that fruit obtains, by directly extracting the prepared product that common kiwi fruit water-soluble concentrate obtains with ethyl acetate, by in distilled water, extracting the prepared product that common kiwi fruit obtains, with by at water, extract the prepared product that obtains in chloroform and the ethyl acetate in turn.

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