CN101890030B - Composite capable of preventing bacteria, viruses, oxidation and pigment deposition - Google Patents
Composite capable of preventing bacteria, viruses, oxidation and pigment deposition Download PDFInfo
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- CN101890030B CN101890030B CN200910027678.8A CN200910027678A CN101890030B CN 101890030 B CN101890030 B CN 101890030B CN 200910027678 A CN200910027678 A CN 200910027678A CN 101890030 B CN101890030 B CN 101890030B
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Abstract
The invention discloses a composite capable of preventing bacteria, viruses, oxidation and pigment deposition, which is characterized by being prepared from the following raw materials in proportion by weight: 0.1-10 of epigallocatechin gallate (EGCG), 0-10 of chlorogenic acid, 0-10 of asiaticoside and 0-10 of ginkgolide. The invention also discloses application of the composite in preparing drugs, health-care food and cosmetics, which are capable of preventing bacteria, viruses, oxidation and pigment deposition. The composite of the invention has favorable safety, is suitable for curing respiratory tract infection, condyloma acuminatum, herpes progenitalis, chromatosis and other diseases, simultaneously has anti-oxidation and skin whitening functions, protects cells and DNA from being damaged, can prevent and cure dementia and chloasma, and also can be used for cancer adjuvant therapy.
Description
Technical field
The invention provides a kind of compositions, particularly a kind of compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, the preparation method that also relates to said composition with and medicine, food, cosmetic use, belong to pharmaceutical sanitary field.
Background technology
EGCG, i.e. epigallocatechin gallate (EGCG), is a kind of main activity and the water-soluble components of extracting from green tea, is also the chief component composition of Green Tea Polyphenols.EGCG has very strong antioxidation, anticancer, mutation isoreactivity.Antioxidant activity is at least ascorbic more than 100 times; 25 times of vitamin E; can Cell protection and DNA undermined; this infringement is believed relevant with other major diseases with cancer, heart disease; these effects of EGCG ascribe their the removing ability to oxygen-derived free radicals to; in addition, it also, as the inversion agent of multi-drug resistance of the tumor, can improve the sensitivity of cancerous cell and chemotherapy and alleviate the toxicity to heart.Much research shows, EGCG can damage by free radical resisting DNA, and radioprotective and ultraviolet stop oil peroxidation, reduce the content of low density cholesterol, extremely-low density cholesterol and triglyceride in serum, the signal transmission that interfere with cancer cells is survived required, suppresses carcinogen in diet, stops the vigor of some carcinogen with other enzymes in intestinal, liver and lung and antioxidant combined effect, remove free radical, resist pollution, the impact of Exposure to Sunlight and smoking, prevents skin aging and wrinkling.
Chlorogenic acid (chlorogenic acid) is a kind of Phenylpropanoid Glycosides class material that plant synthesizes in aerobic respiration process.Be many Chinese herbal medicine as one of main effective ingredient of Flos Lonicerae, the Cortex Eucommiae, Herba Artemisiae Scopariae etc., be also the important activity composition in numerous fruits vegetable.Chlorogenic acid has the multiple efficacies such as the free radical, anti-inflammation, antiviral, blood sugar lowering, blood fat reducing, hepatic cholagogic of removing, anticancer, anti-AIDS.In addition, as a kind of good antioxidant, chlorogenic acid also has application in the field such as daily-use chemical industry, food.
Asiaticoside is active component main in Chinese medicine Herba Centellae, and abundant because of its pharmacologically active, clinical effect is extensive, and application prospect is large, has exploitation and is worth.In recent years, studies confirm that in a large number asiaticoside has antitumor, anti-venous insufficiency, antiulcer, supporting wound healing, antidepressant and recovers the multiple pharmacologically actives such as function of nervous system, also shows the pharmacological action at aspects such as cardiovascular, immunomodulating, antiinflammatory, antiviral simultaneously.
Bilobalide is the main pharmacodynamics position in Folium Ginkgo extract and preparation thereof, is generally acknowledged platelet activating factor (PAF) receptor antagonist.Comprise Ginkgolide A. B. C, J and M (GKA, GKB, GKC, GKJ, GKM) etc.That in bilobalide, anti-PAF is the highest is GKB, is secondly GKA, is GKC again.Along with the drug development arrival in " green waves " epoch, the bilobalide that derives from natural plants receives increasing concern.The main pharmacological of bilobalide has antiplatelet aggregation and thrombosis, prevents and treats atherosclerosis, resisting cardiovascular ischemic injuries, to strengthen neuronic inhibition active, the effects such as epilepsy, anxious, shock, antiallergic, anti-inflammatory, varying cyclically due to the immunoreation of bilobalide to cell, eyes pathological changes, endotoxin is also inhibited, also for multiple diseases fields such as transplant rejection, apoplexy, presenile dementia, septic shock, multiple sclerosis, extracorporeal circulation, cerebral hemorrhage and burns.
Above four kinds of raw materials respectively have feature, be widely used, but in prior art, be also provided with its applied in any combination report at aspects such as medicine, food, cosmetics.
Summary of the invention
Technical problem to be solved by this invention is for the deficiencies in the prior art, and the compositions of a kind of antibacterial, antiviral, antioxidation, anti-pigmentation is provided.
The present invention also provides the purposes of above-mentioned composition.
Technical problem to be solved by this invention is to realize by following technical scheme.The present invention is the compositions of a kind of antibacterial, antiviral, antioxidation, anti-pigmentation, is characterized in, it is made up of at least one in epigallocatechin gallate (EGCG) and chlorogenic acid, asiaticoside and bilobalide, and its weight proportion is as follows:
Epigallocatechin gallate (EGCG) 0.1～10
Chlorogenic acid 0～10
Asiaticoside 0～10
Bilobalide 0～10.
Technical problem to be solved by this invention can also further realize by following technical scheme.The compositions of above-described antibacterial, antiviral, antioxidation, anti-pigmentation, is characterized in, it is made up of at least one in epigallocatechin gallate (EGCG) and chlorogenic acid, asiaticoside and bilobalide, and its weight proportion is as follows:
Epigallocatechin gallate (EGCG) 0.1～10
Chlorogenic acid 0.1～10
Asiaticoside 0.1～10
Bilobalide 0.1～10.
In above-described composite formula of the present invention, epigallocatechin gallate (EGCG) is the indispensable raw material of prescription; Chlorogenic acid, asiaticoside and bilobalide are optional raw materials; In 3 kinds of optional raw materials, at least one can carry out prescription with epigallocatechin gallate (EGCG), that is: can be in chlorogenic acid, asiaticoside and bilobalide any a kind with epigallocatechin gallate (EGCG) prescription, form the compositions of 2 taste raw material prescriptions; Also can be any 2 kinds and epigallocatechin gallate (EGCG) prescription in chlorogenic acid, asiaticoside and bilobalide, form the compositions of 3 taste raw material prescriptions; Can also be chlorogenic acid, asiaticoside and 3 kinds of bilobalides and epigallocatechin gallate (EGCG) prescription, form the compositions of 4 taste raw material prescriptions, 4 taste raw material prescriptions be preferred feedstock prescription; In the time that weight proportion in optional raw material is selected " 0 ", represent and in prescription, do not contain this raw material.
In each raw material of the present invention: epigallocatechin gallate (EGCG) is extraction from Folium Camelliae sinensis, refines the polyphenol component of coming, and purity is more than 50%, preferably more than 90%; Chlorogenic acid be rich in the plant of chlorogenic acid from Flos Lonicerae, Folium Eucommiae etc. extract, refining come phenolic acid composition, purity is more than 50%, preferably more than 90%; Asiaticoside is extraction from Herba Centellae, refines the saponin constituent of coming, and in total saponins or asiaticoside, asiaticoside and Asaiticoside B, at least one component content is more than 50%, preferably more than 70%; Bilobalide is from Semen Ginkgo, Folium Ginkgo, to extract, refine the Ginkgolide Component of coming, and in total lactone or Ginkgolide A. B. C, J, M, K, L and bilobalide, at least one component content is more than 50%, preferably more than 90%.
Active ingredient raw material of the present invention can be commercially available epigallocatechin gallate (EGCG), chlorogenic acid, asiaticoside, bilobalide, also can be the active ingredient that extracting method extracts gained from corresponding plants routinely, can be also the active ingredient that in prior art, disclosed any extracting method extracts.
The purposes of the present composition is the application in antibacterial, the antiviral of preparation, antioxidation, anti-pigmentation medicine, health food, cosmetics; its function comprises the diseases such as control respiratory tract infection, condyloma acuminatum, genital herpes, pigmentation; there is antioxidation simultaneously; Cell protection and DNA are undermined, effect of whitening; can prevent and treat dementia, chloasma, also can be used for the auxiliary treatment of cancer.
Extract of the present invention can be prepared into various medicines, health food, cosmetics, and its included peroral dosage form has capsule, soft capsule, sheet, drop pill, granule, oral liquid, effervescent tablet etc.Exterior-applied formulation comprises tincture, unguentum, gel, patch, membrane, cream, cataplasma, lotion, liniment, paste, aerosol, spray, rubber-emplastrum or liniment.The required adjuvant of preparation can be disclosed any medicinal, edible adjuvant applicatory in prior art, comprises required suitable substrate or excipient, and preparation method can be taked disclosed any method applicatory in prior art according to its dosage form.
The present composition, through a large amount of basic ratio optimizations, screening study, has been determined preferably part combination and good functional activity, and each prescription raw material all has certain Synergistic function.It is below the drug action experiment that inventor does.
Experiment one, the research of this compositions different formulations antibacterial activity.
Precise quantitative takes each proportioning compositions, and (EGCG is alone; EGCG+ chlorogenic acid 5: 5; EGCG+ chlorogenic acid+asiaticoside 5: 5: 5; EGCG+ chlorogenic acid+asiaticoside+bilobalide 5: 5: 5: 5; EGCG+ asiaticoside 5: 5; EGCG+ bilobalide 5: 5; EGCG+ asiaticoside+bilobalide 5: 5: 5; ), be mixed with the solution of 20mg/ml with sterilized water.Then the solution of preparation is carried out to doubling dilution with appropriate sterilized water respectively, make each medicinal liquid become a series of Concentraton gradient, successively: 20,10,5,2.5,1.25,0.625,0.312,0.156,0.078,0.039 (mg/ml).Then get each gradient medicinal liquid 2ml, join respectively in aseptic plate (plate diameter 9cm), then the MH culture medium 18ml of insulation in 55 DEG C of water-baths that adds sterilizing to melt, fully mix immediately, stand-by after condensation.Establish blank simultaneously, only add the agar culture medium of 20ml.
Preactivated each test strain is inoculated into respectively in 2ml culture fluid, 37 DEG C of overnight incubation, then do suitable dilution with corresponding culture fluid, bacterial concentration is about: 10
7～10
8(CFU/ml) (control bacterial concentration method: by the optical density of spectrophotometric determination culture fluid, then do suitable dilution).Then inoculate the each test organisms of instrument dibbling on the MH of each pastille culture medium flat plate with multiple spot, blank carries out simultaneously.Every some bacteria containing amount is about: 10
4～10
5(CFU).37 DEG C, cultivate 24 hours, observe and record the minimum inhibitory concentration MIC value of each bacterium, and calculate MIC
50and MIC
90.The results are shown in Table 1～7.
The mensuration (MIC:mg/ml) of the alone MIC value to pathogenic bacteria of table 1EGCG
The mensuration (MIC:mg/ml) of the MIC value of table 2EGCG+ chlorogenic acid compositions to pathogenic bacteria
The mensuration (MIC:mg/ml) of the MIC value of table 3EGCG+ chlorogenic acid+asiaticoside compositions to pathogenic bacteria
The mensuration (MIC:mg/ml) of the MIC value of table 4EGCG+ chlorogenic acid+asiaticoside+bilobalide compositions to pathogenic bacteria
The mensuration (MIC:mg/ml) of the MIC value of table 5EGCG+ asiaticoside to pathogenic bacteria
The mensuration (MIC:mg/ml) of the MIC value of table 6EGCG+ bilobalide compositions to pathogenic bacteria
The mensuration (MIC:mg/ml) of the MIC value of table 7EGCG+ asiaticoside+bilobalide compositions to pathogenic bacteria
The above results shows that each combined sample all has certain antibacterial activity to each test strain as Klebsiella pneumoniae, staphylococcus aureus, Serratieae, staphylococcus epidermidis, salmonella, escherichia coli, aerobacteria, citrobacter, Bacillus proteus, proteus mirabilis, bacillus pyocyaneus, enterobacter cloacae, shigella, Hemolytic streptococcus, acinetobacter calcoaceticus, streptococcus pneumoniae, bloodthirsty hemophilus influenza etc.And each EGCG prescription effect is all better than EGCG folk prescription effect.
Experiment two, this compositions different formulations are to anti-influenza A virus, Influenza B virus research.
(1) virus virulence is measured:
Cell is by 1 × 10
5concentration, be inoculated on 96 porocyte culture plates, every hole 0.2ml, 24h grows up to cell monolayer, supernatant discarded, by different virus dilution ratios (10
0, 10
-1, 10
-2, 10
-3, 10
-4, 10
-5), add 20 μ L virus liquids, 4 multiple holes of each concentration, absorption 1h, washes away the virus of not adsorbing with Hank ' s solution, add cell culture fluid 1ml, 37 DEG C of 5%CO
2cultivate 72h.According to occurring cytopathy situation, calculate viral TCID with Reed-Munch formula
50.
Result shows: the TCID of influenza A virus
50=10
-3.3, the TCID of Influenza B virus
50=10
-4.6, the TCID of adenovirus
50=10
-4.8.
(2) toxic action of compositions to cell
Mdck cell is pressed to 1 × 10
5concentration, be inoculated on 96 porocyte culture plates, every hole 0.2ml, grows up to cell monolayer after 24h, for subsequent use.Supernatant discarded, will containing different proportioning compositionss, (EGCG be alone; EGCG+ chlorogenic acid 5: 5; EGCG+ chlorogenic acid+asiaticoside 5: 5: 5; EGCG+ chlorogenic acid+asiaticoside+bilobalide 5: 5: 5: 5; EGCG+ asiaticoside 5: 5; EGCG+ bilobalide 5: 5; EGCG+ asiaticoside+bilobalide 5: 5: 5) the cell culture fluid 0.2ml of concentration adds cell, 4 multiple holes of each concentration, 37 DEG C of 5%CO
2cultivate 3d; Utilize mtt assay to measure cell viability, carry out antivirus test.
Result shows: EGCG is alone, the cytotoxicity maximal non-toxic concentration of EGCG+ chlorogenic acid, EGCG+ chlorogenic acid+asiaticoside, EGCG+ chlorogenic acid+asiaticoside+bilobalide, EGCG+ asiaticoside, EGCG+ bilobalide, EGCG+ asiaticoside+bilobalide compositions is respectively 200,150,150,120,200,200,150mg/ml.
(3) compositions antiviral activity
Mdck cell is pressed to 1 × 10
5concentration, be inoculated on 96 porocyte culture plates, every hole 0.2ml, grows up to cell monolayer after 24h, for subsequent use.Cell blank contrast is established in experiment; Virus control; EGCG is alone; EGCG+ chlorogenic acid 5: 5; EGCG+ chlorogenic acid+asiaticoside 5: 5: 5; EGCG+ chlorogenic acid+asiaticoside+bilobalide 5: 5: 5: 5; EGCG+ asiaticoside 5: 5; EGCG+ bilobalide 5: 5; EGCG+ asiaticoside+bilobalide 5: 5: 5; Totally 9 groups.Get the cell that grows up to monolayer, supernatant discarded, adds 100 μ L100TCID
50virus liquid, absorption 1h, washes away the virus of not adsorbing with Hank ' s liquid, add the cell culture fluid 200 μ l containing variable concentrations medicine, 4 multiple holes of each concentration, 37 DEG C of 5%CO
2cultivate 72h.Utilize mtt assay to measure cell viability.The results are shown in Table 8,9.
Mtt assay is measured cell viability: finishing front 4h in experiment, to add final concentration be the MTT of 0.5g/L, DEG C, 5%CO
2abandoning supernatant after lower effect 4h, every hole adds DMSO150 μ L, puts the 10min that vibrates on agitator crystallization is fully dissolved, and measures OD value, observation of cell vigor with microplate reader 490nm.
Mtt assay result shows, each compositions group is to influenza A virus H
1n
1strain all has certain inhibitory action, compares with virus control group in certain concentration range, has significant difference (P < 0.01, P < 0.05).
Each compositions group all has obvious inhibitory action to influenza B virus strain, and OD value is compared with virus control group, has remarkable significant difference (P < 0.05).
The inhibitory action (X ± SD) of table 8 compositions different formulations to influenza A virus
Note: compare with virus control group,
*p < 0.01,
*p < 0.05
The inhibitory action (X ± SD) of table 9 compositions different formulations to Influenza B virus
Note: compare with virus control group,
*p < 0.05,
*p < 0.01.
Experiment three, this compositions different formulations antagonism herpes simplex virus II type activity research
Herpes simplex virus (HSV) is a kind of common infectious agent, and HSV-II can cause herpes vulvaris, is the collaborative inducement of cervical cancer.And at present clinical conventional antiviral drugs often specificity is not strong, or because of effects limit such as easily generation drug resistance strain and side effect application.This experiment is intended to, by the activity of the anti-herpes simplex virus II type of this compositions of in vitro study different formulations, inquire into its therapeutical effect to viral genital system infection disease.
(1) virus titer is measured
Viral suspension is done to continuous 10 times of dilutions, and conventional method infects monolayer Vero cell, 37 DEG C, 5%CO
2cultivate 3d, observation of cell pathological changes situation, calculates TCID by Reed-Muench method
50.
(2) cytotoxic assay of sample
The take the logarithm cell of trophophase, after centrifugal (1000r/min, 10min), cell is resuspended, cell concentration is adjusted to 2 × 10
6/ ml.On 96 porocyte culture plates, by 2 times of doubling dilutions of test sample, establish altogether 6 gradients, each gradient is established 3 repeating holes, separately establishes positive controls and cell matched group.Add respectively after cell suspension, put 37 DEG C, 5%CO
2in incubator, hatch.Measured the OD value in each hole by mtt assay microplate reader in the 4th day.Calculate the suppression ratio of test sample cell growth by following formula, and calculate IC
50(50%inhibition concentration value).
Inhibitory rate of cell growth (%)=(1-experimental port OD value/control wells OD value) × 100%
(3) protective effect of mtt assay detection compound to herpesvirus infection Vero cell
On the 96 flat culture plates in hole, by testing combination, (EGCG is alone; EGCG+ chlorogenic acid 5: 5; EGCG+ chlorogenic acid+asiaticoside 5: 5: 5; EGCG+ chlorogenic acid+asiaticoside+bilobalide 5: 5: 5: 5; EGCG+ asiaticoside 5: 5; EGCG+ bilobalide 5: 5; EGCG+ asiaticoside+bilobalide 5: 5: 5) carry out 2 times of serial dilutions by RPMI1640 culture medium.Totally 6 dilution factors, every hole 100 μ l, establish two repeating holes.The cell matched group of the uninfecting virus that does not contain compositions is set simultaneously and infects viral virus control group; Centrifugal Vero cell (1000r/min, 10min), uses complete medium re-suspended cell, and every hole adds 75 μ l 4 × 10
4cell; Two holes in each dilution factor of infection HSV add 25 μ l viruses, and matched group adds 25 μ l culture medium.Every hole final volume 200 μ l.Put 37 DEG C, 5%CO
2in incubator, cultivate, within the 3rd day, add the fresh training base of 100 μ l; Within the 6th day, by after 96 porocyte culture plate low-speed centrifugals (1000r/min, 10min), 250 μ l supernatants are abandoned in suction, then add 20 μ lMTT solution (5mg/ml), 37 DEG C, 5%CO
2in incubator, hatch 4h left and right, add 20%SDS-50%DMF (pH 4.7), overnight incubation; Within the 7th day, measure the OD value in each hole by microplate reader, calculate the protective rate of compositions to HSV-II infection cell according to formula below, and calculate EC by insertion
50(50%Effective concentration) value and therapeutic index TI (Therapeutic index) value:
And further calculate therapeutic index (TI=IC
50/ EC
50).
The titre that HSV-II is seeded on Vero cell is 103.0/ml.Experiment is 80TCID with virus concentration
50.
The results are shown in Table 10.
The protective effect of this compositions of table 10 different formulations to II herpes simplex virus type vero cells infection
Result shows: positive control AVC demonstrates the effect of protection HSV-II vero cells infection, its EC
50be that 5.33 μ g/ml therapeutic index TI values are 2814.The EC of the each formula group of this compositions
50value and therapeutic index are in Table.Compared with positive control medicine ACV, the therapeutic index (TI) of the each formula group of this compositions although on the low side, HSV-II vero cells infection has still been demonstrated to stronger protective effect.
The effect research of experiment four, this compositions different formulations anti-human papilloma virus (anti-HPV)
It is the Etiological that causes condyloma acuminatum (CA) that human papillomavirus (HPV) infects, for inquiring into the effect of this compositions different formulations anti-human papilloma virus (anti-HPV) effect, we adopt fluorescence quantitative polymerase chain reaction (FQ-PCR) technology, observe the impact of the HPV-DN of EGCG in vitro lesions of condyloma acuminatum, find that this compositions different formulations has the effect of anti-human papilloma virus (anti-HPV).
(1) preparation of CA suspension
Come from the wart body specimen (through being accredited as HPV6/11 type) at patients with condyloma acuminatum pudendum position from the department of dermatologry collection of the first Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, focus in a centrifuge tube, 30mg weighs, use glass homogenizer homogenate, add again 5ml normal saline, be made into the in vitro CA suspension of 6mg/ml, for subsequent use.
(2) method of sample treatment wart body
Draw 25 μ l sample liquid and 25 μ l CA suspensions to 1.5ml centrifuge tube with liquid-transfering gun, jolting is even, and (EGCG is alone to make the each formula group of this compositions; EGCG+ chlorogenic acid 5: 5; EGCG+ chlorogenic acid+asiaticoside 5: 5: 5; EGCG+ chlorogenic acid+asiaticoside+bilobalide 5: 5: 5: 5; EGCG+ asiaticoside 5: 5; EGCG+ bilobalide 5: 5; EGCG+ asiaticoside+bilobalide 5: 5: 5) (final concentration is 100ug/ml) and wart physical ability fully act on, and puts 37 DEG C of water baths and cultivate, and separately establishes normal saline group, positive control (acyclovir) group, distilled water matched group., after 1,3,5,7 days, respectively get one group and carry out following test at sample treatment wart body.
(3) extraction of HPV-DNA
From warm bath cabinet, take out through the CA of sample treatment suspension, extract DNA by test kit step: add 50 μ l normal saline, after mixing, add again 100 μ l DNA extraction liquid; Jolting is even, the centrifugal 10min of 1200r/min, and abandoning supernatant, then add 25 μ l DNA extraction liquid; Fully mix, 100 DEG C of boiling water boiling 10min, the centrifugal 10min of 1200r/min, supernatant is HPV-DNA template.
(4) DNA cloning
Get the PCR reactant liquor 37.6 μ l in detection kit, Taq archaeal dna polymerase 0.4 μ l, UNG 0.03 μ l, in PCR reaction tube, then adds HPV-DNA template 2 μ l, and button strict control lid, is placed in quantitative PCR instrument cocycle amplification.HPV-DNA carries out cyclic amplification through high-temperature denatured, process annealing and extension, and cyclic program is set to: 37 DEG C, and 5min; 94 DEG C, 1min; 95 DEG C, 5sec; 60 DEG C, 30sec; Circulate 40 times.
(5) quantitative criterion curve
Quantitative with fluorescent probe detection, according to detection kit requirement, set up the quantitative positive criteria product of HPV-DNA of 4 series concentration, be followed successively by 5 × 10
7/ ml, 5 × 10
6/ ml, 5 × 10
5/ ml, 5 × 10
4/ ml, through PCR detection, (its detection sensitivity is 1 × 10
3/ ml, result is less than this concentration person and is judged to be feminine gender).Taking the natural logrithm of initial copy number as abscissa, cycle threshold is vertical coordinate, and the regression straight line obtaining is standard curve, accordingly the amplification times of sample is carried out quantitatively.
Positive findings taking the quantitative logarithm value of DNA cloning represent (unit as: copy/mL), illustrate that virus is not damaged, its DNA is able to a large amount of amplifications; Negative findings represents with "-", shows that viral DNA is subject to killing destruction, increases suppressed.Carry out 3 tests according to above step, measure the amplification quantity of DNA and average.The results are shown in Table 11.
This compositions of table 11 different formulations is to HPV-DNA exercising result (copy/ml, n=3)
As can be known from the above table: normal saline and distilled water group result are positive, HPV-DNA quantity after pcr amplification reaction can reach 10
7, visible virus replication and survival ability are extremely strong; And through the each formula effect of this compositions after 1 day, PCR result is negative, this compositions different formulations is described, and it has obviously broken ring effect to virus, suppress it to copy amplification, and effective drug duration is longer; Solvent is the action effect of interference medicament not.
Experiment five, the impact of this compositions different formulations on Cavia porcellus genital herpes
90 Cavia porcelluss are got to vaginal secretions in advance drips plate and detects without after HSV-2 viral infection, be divided into immediately 6 groups: normal saline matched group, alone group of EGCG, EGCG+ chlorogenic acid group (5: 5), EGCG+ chlorogenic acid+asiaticoside (5: 5: 5) group, EGCG+ chlorogenic acid+asiaticoside+bilobalide group (5: 5: 5: 5), EGCG+ asiaticoside (5: 5), EGCG+ bilobalide (5: 5), EGCG+ asiaticoside+bilobalide (5: 5: 5), acyclovir (ACV) group, 10 every group.Normal saline cleans pudendum, and dry cotton swab friction vagina for several times, is then kowtowed gently agouti pudendum with percussopunctator and made its cunnus hyperraemia and a little oozing of blood, and clean cotton swab is cleaned pudendum.Draw 0.15ml HSV-2 Strain with 1ml syringe, syringe needle is stretched into the about 3-4cm of Cavia porcellus intravaginal after mixing gavage syringe needle, slowly exit after virus is injected to fornix vagina, fill in vagina to maintain venom with gelfoam.On syringe needle, a little venom drops in pudendum, smears gently evenly with glass rod, makes virus go deep into skin.
Symptom score method: mark according to lesion degree: 0 is asymptomatic; 0.5 for only red and swollen without vesicle; 1.5 is single phlysis (≤2mm); 2.0 is single large vesicle (> 2mm); 2.5 is that multiple phlysises or ulcer of vagina are hemorrhage; 3.0 is multiple large vesicles; 3.5 is serious swelling of external genitals; 4.0 is that multiple large phlyctens merge; 4.5 is rear acroparalysis; 5.0 is vulval ulcer.
Medication: acyclovir group group Cavia porcellus is given and 0.0775g/kg/d respectively with before modeling, after gavage 5d, according to modeling method virus inoculation 0.15ml, again, after successive administration 10d, observes Cavia porcellus whole body and pudendum symptom every day.The each recipe ingredient of this compositions does not dip with cotton pellet the solution 0.1ml that concentration is 0.1g/ml, 0.05g/ml after modeling 4h, draw the each concentration of 0.1ml with syringe and be subject to test product, after mixing gavage syringe needle, syringe needle is stretched into the about 3-4cm of Cavia porcellus intravaginal, after being injected to fornix vagina, virus slowly exits, on syringe needle, a little solution drops in pudendum, smear gently evenly with glass rod, make virus go deep into skin.Within the 1st, 3,5,7,9 days after virus inoculation, respectively organize Cavia porcellus pudendum with 0.1ml normal saline flushing respectively, get and rinse drop plate mensuration virus titer.The results are shown in Table 12,13.
The impact (x ± s, n=10) of this compositions of table 12 different formulations on Cavia porcellus intravaginal virus titer
*represent P < 0.05,
*represent P < 0.01
As seen from the above table, after the inoculation of normal saline group, the 1st day infection titer is the highest, extends gradually and declines in time later.Other administration groups are along with the increase of administration natural law, virus titer is downward trend gradually substantially, wherein obvious with the downward trend of acyclovir group, and the 3rd, 7,9d and blank group relatively have utmost point significant difference (P < 0.01), within the 5th day, has significant difference (P < 0.05); Simultaneously each group of this compositions different formulations relatively has significant difference (P < 0.05) at administration the 5th, 7,9d virus titer with blank group, has shown good antiviral effect.
The impact (x ± s, n=10) of table 13EGCG on Guinea Pig Model of Genital Herpes pudendum clinical symptoms
*represent P < 0.05,
*represent P < 0.01
As seen from the above table, all there is being similar to the symptom of mankind's pudendum genital herpes in the Cavia porcellus after virus inoculation, and main manifestations is that pudendum is red, swollen, vesicle, ulcer, incrustation.There is vesicle in the most 2d of normal saline matched group, 5～7d pathological changes is the most obvious, and later symptom alleviates gradually.There is vesicle in the most 3～5d of the each group of this compositions different formulations, and symptom score is all lower than normal saline matched group.Between pudendum symptom score group, relatively, there were significant differences (P < 0.05, P < 0.01).
Experiment six, evaluate the oxidation resistance of this compositions different formulations by removing organic free radical DPPH method
This experiment is inquired into the oxidation resistance of composite formula from external removing free radical ability aspect, to further verify its oxidation resistance.
1, the preparation of DPPH free radical sample solution
Accurately weighed a certain amount of DPPH, is dissolved in 95% ethanol, after again with the fixed molten 1000ml that causes of 95% ethanol, making the ultimate density of DPPH is 38.60mg/L, keeps in Dark Place.
2, the preparation of this compositions different formulations solution example
Accurate weighed this compositions of the 400.00mg different formulations of decrement method is subject to test product, and with the fixed molten 1000ml that causes of 95% ethanol, making final concentration is 400mg/L.
3, the preparation of bad hematic acid solution example
The accurate weighed bad hematic acid of 400.00mg of decrement method, with the fixed molten 1000ml that causes of 95% ethanol, making its final concentration is 400mg/L.
4, this compositions different formulations is removed the testing procedure of free radical ability
(1) get the DPPH free radical sample solution that a teat glass wherein adds 5ml, add again equivalent 95% alcoholic solution, mix homogeneously, 37 DEG C of lucifuge reaction 1h, the zeroing taking 95% ethanol as blank tube, measure its absorbance at 517nm with ultraviolet spectrophotometer, a same sample replication three times, averages and is designated as A0.
(2) teat glass that separately amount of trying to please is 10ml, every pipe precision adds this compositions different formulations solution example of 1ml, 2ml, 3ml, 4ml, 5ml successively in order, then every pipe adds the DPPH free radical sample solution of 5ml again, and the part of not enough 10ml is supplied with 95% ethanol.Each test tube evenly mixes, after 37 DEG C of lucifuges reaction 1h with, 95% ethanol is blank tube zeroing, measures its absorbance at 517nm with ultraviolet spectrophotometer, with a sample replication three times, averages and records respectively numerical value.
(3) repeat step, in test tube, application of sample replaces and is subject to test product solution example with bad hematic acid solution, and application of sample volume and order are the same, result record.
(4) teat glass that separately amount of trying to please is 10ml, every pipe precision adds this compositions different formulations solution example of 1ml, 2ml, 3ml, 4ml, 5ml successively in order, then every pipe is supplied and is caused 10ml with 95% alcoholic solution, each test tube evenly mixes, after 37 DEG C of lucifuges reaction 1h with, 95% ethanol is blank tube zeroing, measures its absorbance at 525nm with ultraviolet spectrophotometer, a same sample replication three times, averages and records respectively numerical value.
(5) repeat step, in test tube, application of sample replaces tested solution example with bad hematic acid solution, and application of sample volume and order are the same, result record.
(6) twice of repeating step (1)～(5), each result record respectively.
5, date processing and calculating
Clearance rate according to above testing result calculation sample to DPPH free-atom aqueous solution, computing formula is as follows: clearance rate=[1-(A
n-B
n)/A
0] × 100%
SPSS 13.0 statistical softwares processing for result of calculation, result represents with x ± s, the results are shown in Table 14.
Table 14DPPH method is evaluated the external removing free radical of this compositions different formulations capability result (x ± s)
From upper table result, this compositions different formulations has very strong removing free radical ability, and under same concentrations, it is removed free radical ability and is better than bad hematic acid.
Antioxidant activity research in experiment seven, this compositions different formulations body
Buy after animal under laboratory environment normal experiment three days, after be divided into immediately 9 groups: 5), EGCG+ asiaticoside (5: 5), EGCG+ bilobalide (5: 5), EGCG+ asiaticoside+bilobalide (5: 5: 5) blank group, model control group, alone group of EGCG, EGCG+ chlorogenic acid group (5: 5), EGCG+ chlorogenic acid+asiaticoside (5: 5: 5) group, EGCG+ chlorogenic acid+asiaticoside+bilobalide group (5: 5: 5:, female half and half, 10 every group.Wherein the other gavage of each recipe ingredient gives the test sample solution that is subject to of 0.5g/kg, and administration volume is 20ml/kg, and two groups, blank, model awards isometric normal saline, and be administered once every day, altogether administration 30d.Last administration evening on the same day, water 8h was can't help in the fasting of each group mice, and rear difference gastric infusion 1 time again, after 30 points of administrations, except blank group, respectively organize gavage and give 0.4mg/kg bromobenzene oil, gavage amount 10ml/kg, after 18 hours, put to death animal, get serum, measure its SOD and GSH-Px level by test kit explanation.Experimental result is in table 15.
This compositions of table 15 different formulations causes mice serum oxidation resistance to bromobenzene oil and reduces model
The impact of SOD in serum and GSH-Px (x ± s)
△ △represent and relatively P < 0.01 of blank group,
*represent and relatively P < 0.01 of model group
As seen from the above table: after modeling, in model group mice serum, SOD and GSH-Px level obviously lower (P < 0.01), modeling success is described.In this compositions different formulations administration group mice serum, SOD raises compared with model group with GSH-Px level obviously (P < 0.01), illustrates that this compositions different formulations has oxidation resistance in stronger body.
Experiment eight, the research of this compositions different formulations anti-experimental character pigmentation
1, cutaneous pigmentation animal model, experiment grouping
Get 90 of Cavia porcelluss, female, body weight 230～250g, gets 10 as normal group, and all the other are divided into 5 groups at random.Grouping: (1) normal group: give excipient; (2) model group: give excipient; (3) alone group of EGCG: 0.25g/kg; (4) EGCG+ chlorogenic acid (5: 5) group: 0.25g/kg; (5) EGCG+ chlorogenic acid+asiaticoside (5: 5: 5) group: 0.25g/kg; (6) EGCG+ chlorogenic acid+asiaticoside+bilobalide group (5: 5: 5: 5): 0.25g/kg; (7) EGCG+ asiaticoside (5: 5) 0.25g/kg; (8) EGCG+ bilobalide (5: 5) 0.25g/kg; (9) EGCG+ asiaticoside+bilobalide (5: 5: 5) 0.25g/kg.Except normal group, all the other ultraviolet B radiations (UVB) of respectively organizing the about 320nm of Cavia porcellus wavelength irradiate Cavia porcellus once every day, each irradiation time 60min, continuous 30 days.Modeling administration every day simultaneously.
2, ordinary circumstance is observed
Observe ordinary circumstance and depilation position, the back change of skin situation of Cavia porcellus every day.
3, dermal melanin cell pathology morphological observation, melanin Positive Objects computer image analysis
After last coating 24h, de-cervical vertebra is put to death Cavia porcellus, strips one of back, Cavia porcellus right side skin of unhairing, fixes with 10% formalin.Routine paraffin wax embedded section, thick 4～5 μ m of sheet, microscope slide drags for sheet, dries sheet after anti-flake is processed.Again through dewaxing to water, antigen retrieval.Use HMB
45(melanoma melanoma) is first antibody, with Elivision
tMplus ImmunohistochemistryMethods Methods shows the melanin granule in epidermis, to evaluate distribution and the secretory function situation of melanocyte in epidermis.
Every animal unhairing position skin of each group is made 1 pathological section, observe 5 different visuals field for every, find after Positive Objects, target image is carried out to quantitative analysis with pathological image analyser: draw the target area, target number, average gray, average optical, integral optical density in 5 visuals field of every section etc.
4, experimental result
Ordinary circumstance:
Ultraviolet radiation is observed modeling Cavia porcellus depilation position change of skin situation after 20 days, find that each modeling group depilation position skin colour is secretly in normal group, and black speckle appears in Some Animals skin day by day, the most serious with model group, and each administration group situation is better than model group.
Pathological examination:
Normal group epidermis is stratified squamous epithelium, and basal layer cell is cube or column, the cell of visible indivedual melanin granule stained positive in it.The visible hair follicle of corium, sebaceous gland and hair, upper subcutaneous be fine and close fibrous connective tissue.Have no the cell of melanin granule stained positive.
Model group epidermal structure is normal, looks into and see that volume kytoplasm dyes dark tan cell under low power lens in basal layer, sees the granule that is full of the immunohistochemical staining positive in cell under high power lens, because amounts of particles is many, causes kytoplasm to be dark brown brown.Cell quantity is many, normal intensive embarking on journey.
Alone group of EGCG: look in basal layer and see that more kytoplasm dyes dark tan cell, cell quantity reduces compared with model group, and staining power weakens.
EGCG+ chlorogenic acid group: basal layer endoplasm dyes dark tan melanocyte quantity to be reduced compared with model group, and most regions melanocyte is dispersed in distribution.
EGCG+ chlorogenic acid+asiaticoside group: basal layer endoplasm dyes dark tan melanocyte quantity obviously to be reduced, melanocyte be dispersed in, sparse distribution, dye shallow.
EGCG+ chlorogenic acid+asiaticoside+bilobalide group: skin basal layer melanocyte quantity obviously reduces, and melanocyte is dispersed in distribution, seldom dense arrangement.In corium and upper subcutaneous tissue, do not look into and see melanocyte.
EGCG+ asiaticoside group: melanocyte quantity reduces compared with model group, most of region melanocyte is dispersed in distribution.
EGCG+ bilobalide group: the melanocyte quantity of basal layer endoplasm reduces.
EGCG+ asiaticoside+bilobalide group: basal layer endoplasm dyes dark tan melanocyte quantity to be reduced compared with model group, and melanocyte sparse distribution, dyes more shallow.
On the impact of modeling Cavia porcellus melanocyte area, quantity
Model group Cavia porcellus melanocyte target area after modeling, target number, apparently higher than normal group (p < 0.05, p < 0.01); Each administration group Cavia porcellus melanocyte index is starkly lower than model group, points out the each formula group of this compositions to have certain inhibitory action to the increase of model melanin area, quantity.The results are shown in Table 16.
The impact (X ± S) of this compositions of table 16 different formulations on modeling Cavia porcellus melanocyte area, quantity
Note: with normal group comparison
△ △p < 0.01; With model group comparison
*p < 0.05,
*p < 0.01.
On the impact of modeling Cavia porcellus melanocyte dyeing weight
Melanocyte dyed color is darker, and gray value is less, and optical density is larger.Integral optical density is the product of average optical and area, and color is darker, and area is larger, and integral optical density is higher.After modeling, Cavia porcellus melanocyte average gray value obviously reduces, average optical density value, integral optical density value obviously raise (p < 0.01); Compositions is respectively organized average gray value and is raise, average optical density value, integral optical density value reduce, relatively there is significant difference (p < 0.05 with model group, p < 0.01), point out each composite formula group staining cell color obviously thin out compared with model group.The results are shown in Table 17.
The impact (X ± S) of this compositions of table 17 different formulations on modeling Cavia porcellus melanocyte dyeing weight
Note: with normal group comparison
△ △p < 0.01; With model group comparison
*p < 0.05,
*p < 0.01.
The above results points out the each formula group of this compositions to have certain inhibitory action to the increase of model melanin area, quantity, color.
Through experiment showed, that above the present composition has definite antibacterial, preventing respiratory and infects the effects such as genitals virus, antioxidation, removing free radical, anti-pigmentation such as virus, anti-condyloma acuminatum, genital herpes.
Detailed description of the invention
Below further describe concrete technical scheme of the present invention, so that those skilled in the art understands the present invention further, and do not form the restriction to its right.
Embodiment 1.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid 0.1.Add conventional adjuvant, make capsule or soft capsule, oral for preventing and treating respiratory tract infection.
Embodiment 2.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid 0.1; Asiaticoside 0.1.Add conventional adjuvant, make tablet, the oral auxiliary treatment for cancer.
Embodiment 3.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid 0.1; Asiaticoside 0.1; Bilobalide 0.1.Add conventional adjuvant, make drop pill, oral for preventing and treating presenile dementia.
Embodiment 4.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Chlorogenic acid 1; Bilobalide 0.1.Add conventional adjuvant, make cream, external prevents and treats skin aging and pigmentation.
Embodiment 5.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Chlorogenic acid 1; Asiaticoside 0.1.Add conventional adjuvant, make gel, the anti-treating pointed condyloma of external, genital herpes etc.
Embodiment 6.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Asiaticoside 1; Bilobalide 1.Add conventional adjuvant, make drop pill, oral for treatment of vascular dementia.
Embodiment 7.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 2; Bilobalide 1.Add conventional adjuvant, make unguentum, external prevents and treats chloasma.
Embodiment 8.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.5; Asiaticoside 0.5.Add conventional adjuvant, make membrane, external prevents and treats skin aging.
Embodiment 9.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Chlorogenic acid 5; Asiaticoside 5; Bilobalide 5.Add conventional adjuvant, make lotion, external prevents and treats pigmentation.
Embodiment 10.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Chlorogenic acid 1; Asiaticoside 5; Bilobalide 5.Add conventional adjuvant, make effervescent tablet, oral for preventing and treating aging.
Embodiment 11.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Chlorogenic acid 1; Asiaticoside 5.Add conventional adjuvant, make capsule, oral for preventing and treating viral infection, as influenza.
Embodiment 12.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Chlorogenic acid 1; Asiaticoside 2; Bilobalide 1.Add conventional adjuvant, make vanishing cream, external reaches the object of skin care, whitening.
Embodiment 13.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Chlorogenic acid 10; Asiaticoside 10; Bilobalide 10.Add conventional adjuvant, make drop pill, oral for preventing and treating presenile dementia.
Embodiment 14.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid 10; Asiaticoside 0.1; Bilobalide 10.Add conventional adjuvant, make tablet.
Embodiment 15.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Chlorogenic acid 0.1; Asiaticoside 10; Bilobalide 0.1.Add conventional adjuvant, granulation agent.
Embodiment 16.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Chlorogenic acid 5; Asiaticoside 1; Bilobalide 1.Add conventional adjuvant, make pill.
Embodiment 17.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 2; Chlorogenic acid 8; Asiaticoside 8; Bilobalide 2.Add conventional adjuvant, make drop pill.
Embodiment 18.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 3; Chlorogenic acid 4; Asiaticoside 3; Bilobalide 4.Add conventional adjuvant, make capsule.
Embodiment 19.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Chlorogenic acid 7; Asiaticoside 2; Bilobalide 3.Add conventional adjuvant, make cream.
Embodiment 20.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 2; Chlorogenic acid 4; Asiaticoside 8; Bilobalide 7.Add conventional adjuvant, make oral liquid.
Embodiment 21.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid or asiaticoside or bilobalide 10.Add conventional adjuvant, make capsule or soft capsule, oral for preventing and treating respiratory tract infection.
Embodiment 22.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Chlorogenic acid or asiaticoside or bilobalide 0.1.Add conventional adjuvant, make capsule or soft capsule.
Embodiment 23.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Chlorogenic acid or asiaticoside or bilobalide 5.Add conventional adjuvant, make tablet.
Embodiment 24.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Chlorogenic acid or asiaticoside or bilobalide 1.Add conventional adjuvant, make oral liquid.
Embodiment 25.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 3; Chlorogenic acid or asiaticoside or bilobalide 6.Add conventional adjuvant, make capsule.
Embodiment 26.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Chlorogenic acid 0.1; Asiaticoside or bilobalide 10.Add conventional adjuvant, make tablet, the oral auxiliary treatment for cancer.
Embodiment 27.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Chlorogenic acid 10; Asiaticoside or bilobalide 0.1.Add conventional adjuvant, make cream, external prevents and treats skin aging and pigmentation.
Embodiment 28.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 2; Chlorogenic acid 3; Asiaticoside or bilobalide 5.Add conventional adjuvant, make gel, the anti-treating pointed condyloma of external, genital herpes etc.
Embodiment 29.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 1; Asiaticoside 1; Asiaticoside or bilobalide 1.Add conventional adjuvant, make drop pill, oral for treatment of vascular dementia.
Embodiment 30.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 2; Asiaticoside 0.1; Bilobalide 8.Add conventional adjuvant, make unguentum, external prevents and treats chloasma.
Embodiment 31.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Asiaticoside 1; Bilobalide 2.Add conventional adjuvant, make membrane, external prevents and treats skin aging.
Embodiment 32.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 5; Asiaticoside 5; Bilobalide 5.Add conventional adjuvant, make lotion, external prevents and treats pigmentation.
Embodiment 33.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 8; Asiaticoside 6; Bilobalide 4.Add conventional adjuvant, make effervescent tablet, oral for preventing and treating aging.
Embodiment 34.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 0.1; Asiaticoside 10; Bilobalide 10.Add conventional adjuvant, make capsule, oral for preventing and treating viral infection, as influenza.
Embodiment 35.A compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, it is made up of the raw material of following weight proportion, epigallocatechin gallate (EGCG) 10; Asiaticoside 10; Bilobalide 1.Add conventional adjuvant, make vanishing cream, external reaches the object of skin care, whitening.
Claims (8)
1. a compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, is characterized in that, it is made up of the raw material of following weight proportion:
Epigallocatechin gallate (EGCG) 0.1～10;
Chlorogenic acid 0.1～10.
2. a compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, is characterized in that, it is made up of the raw material of following weight proportion:
Epigallocatechin gallate (EGCG) 0.1～10;
Chlorogenic acid 0.1～10;
Asiaticoside 0.1～10.
3. a compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, is characterized in that, it is made up of the raw material of following weight proportion:
Epigallocatechin gallate (EGCG) 0.1～10;
Chlorogenic acid 0.1～10;
Bilobalide 0.1～10.
4. a compositions with antibacterial, antiviral, antioxidation, anti-pigmentation effect, is characterized in that, it is made up of the raw material of following weight proportion:
Epigallocatechin gallate (EGCG) 0.1～10;
Chlorogenic acid 0.1～10;
Asiaticoside 0.1～10;
Bilobalide 0.1～10.
5. the compositions described in claim 1 or 2 or 3 or 4 is in the application of preparing in medicine, health food, cosmetics.
6. application according to claim 5, is characterized in that, said composition during for the preparation of medicine, health food, cosmetics, is also added acceptable adjuvant, substrate or additive in said composition.
7. the application of the compositions described in claim 1 or 2 or 3 or 4 in the medicine of antibacterial, the antiviral of preparation, antioxidation and anti-pigmentation, health food, cosmetics.
8. the application of the compositions described in claim 1 or 2 or 3 or 4 in preparation treatment respiratory tract infection, condyloma acuminatum, genital herpes or Pigmented medicine; this medicine has antioxidation simultaneously; Cell protection and DNA are undermined, the function of whitening, and said composition also possesses as the purposes in the medicine of the auxiliary treatment of control dementia, chloasma, cancer.
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| CN103181943A (en) * | 2011-12-30 | 2013-07-03 | 新昌县冠阳技术开发有限公司 | Honeysuckle flower oil enteric-coated capsules |
| CN103431387B (en) * | 2013-08-24 | 2015-04-01 | 湖南绿洲植物资源开发有限公司 | Whitening and freckle-removing traditional Chinese medicine composition and preparation process thereof |
| CN103479562A (en) * | 2013-09-12 | 2014-01-01 | 严晓燕 | Gingko green tea moisture keeping emulsion |
| CN108685891A (en) * | 2017-04-06 | 2018-10-23 | 四川九章生物科技有限公司 | Purposes of the chlorogenic acid in preparing the injection with skin brightening effect |
| CN113521161A (en) * | 2020-04-16 | 2021-10-22 | 三捷生物科技（北京）有限公司 | Pharmaceutical composition with function of resisting microbial infection and application thereof |
| CN111529434B (en) * | 2020-05-11 | 2021-09-03 | 山东农业大学 | Lip prevention and health care product containing EGCG and application thereof |
| CN112293737A (en) * | 2020-12-01 | 2021-02-02 | 曾晓飞 | Omega 3 and phillyrin combined plant antiviral and anti-inflammatory food supplement |
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Non-Patent Citations (4)
| Title |
|---|
| 张瑜等.表没食子儿茶素没食子酸酯研究进展.《氨基酸和生物资源》.1998,第20卷(第4期),51-54. |
| 茶叶中EGCG功效研究进展;赵丽萍等;《中国农学通报》;20070731;第23卷(第7期);143-147 * |
| 表没食子儿茶素没食子酸酯研究进展;张瑜等;《氨基酸和生物资源》;19981231;第20卷(第4期);51-54 * |
| 赵丽萍等.茶叶中EGCG功效研究进展.《中国农学通报》.2007,第23卷(第7期),143-147. |
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