CN101234140B - Pharmaceutical composition for curing eczema and preparation and application thereof - Google Patents
Pharmaceutical composition for curing eczema and preparation and application thereof Download PDFInfo
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Abstract
The invention provides a drug combination for treating eczema, which is a preparation prepared by the raw materials with the following weight proportion: 1-9 shares of skullcap and 9-1share of gentian. The invention also provides a preparation method and purposes of the drug combination. Two raw materials of skullcap and gentian in the invention are compatible to treat eczema. The animal experiment study proves that the compatibility of skullcap and gentian can greatly enhance the efficacy of clearing damp-heat in the liver gallbladder. The drug combination has great effects on anti-inflammation, antianaphylaxis, itch relieving and bacteriostasis, which has remarkable curative effect for treating cutaneous pruritus, erythema, papule or blisters resulted from acute eczema and dermatitis.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition for the treatment of eczema, particularly, is to be the pharmaceutical composition that feedstock production forms with Radix Scutellariae, Radix Gentianae.Belong to the field of Chinese medicines.
Background technology
Eczema, dermatitis (contact dermatitis, allergic dermatitis, autosensitization dermatitis, infectious eczematoid dermatitis etc.) are clinical modal dermatosiss, wherein see with acute eczema and acute dermatitis especially, be one of emphasis problem of the clinical and basic research of domestic and international dermatosis more.Because primary disease is cardinal symptom with violent pruritus often, many with the erythra form, be easy to ooze out, the course of disease is delayed and tendency of recurrence is arranged is feature, has a strong impact on patient's physical and mental health.Character is identical in broad terms with dermatitis for eczema, it all is allergic skin disease former by multiple difference thereby that cause, the two pathological change all is intercellular edema (sponge formation) and intracellular edema to occur at epidermis, and high dermis has telangiectasis, edema and mononuclearcell to soak into.Therefore used as synonym by a lot of scholars clinically.But eczema often is difficult for finding clear and definite reason, and dermatitis reason is clearer and more definite often, as contact some and stimulate stimulus object or allergin, perhaps take or the external some drugs relevant.Primary disease is modal anaphylaxis dermatosis, is found in any age, and man, woman all can fall ill, asexuality difference, and the city is more than the rural area.According to domestic census report, eczema accounts for 15～25% of department of dermatologry outpatient service amount, and wherein acute eczema accounts for 52.4%.Eczema is considered as a big class disease abroad, comprises: atopic dermatitis, limitation chronic dermatitis, contact dermatitis, hand dermatitis, nummular dermatitis, so that its sickness rate is higher than far away is domestic.So eczema is typical commonly encountered diseases, frequently-occurring disease.
Acute eczema mainly with skin lesion pleomorphism, symmetrical distribution, violent pruritus, tendency humidifying and easily repeatedly outbreak be characteristics, severe patient causes complication such as infection, thus bring big misery to the patient, thereby influence quality of life and work, study, life.Acute eczema can betide any position of health, be common in head, face, ear after, breast, distal limbs and pudendum etc. locate.The clinical characters of acute eczema is first-selectedly on the skin the most intensive point-like erythema and the pimple and the papulovesicle (the substrate flushing of pimple has slight edema) of foxtail millet grain size to occur, and becomes phlysis very soon, and the broken back of bleb forms rotten to the corn and and the incrustation etc. of point-like.Subjective symptoms is violent pruritus, and causalgia is arranged, and Chang Yin scratches or the hot water laundering, causes the further diffusion towards periphery of rotten face, makes the skin lesion indefinite border.If processing is proper, inflammation alleviates, and desquamation occurs, and erythra can disappear in 2～3 weeks, as deals with improperly, and the course of disease prolongs, and easily develops into subacute and casual property eczema.According to site of pathological change, eczema has the title difference of ear eczema, breast eczema, omphalos eczema, eczema scrotum, pudendum eczema, anal eczema, hand eczema, legs eczema.
Doctor trained in Western medicine is removed all suspicious paathogenic factors to the treatment 1. of primary disease.Keep skin clean, avoid various environmental stimulis, as hot water launder, firmly scratch, too much use soap, unsuitable medicine for external use etc.Avoid secondary infection, should avoid overworked and psychentonia simultaneously, avoid foods such as hot, peppery, raw meat, acid.For chronic general eczema patients, remove focus in case of necessity, the treatment systemic disease.2. whole body therapeutic comprises (1) antihistaminic class medicine, as chlorphenamine, Cyproheptadine, hismanal etc.; (2) calm pruritus is used for spiritedness obstacle or poor sleep person, drinks and selects chlorpromazine, perphenazine, miltown, 10% chloral hydrate etc.(3) calcium preparation is slowly injected as 10% calcium gluconate injection vein.(4) vein at violent pruritus seals: 0.25% procaine, 10～20ml adds vitamin C 1～2g, intravenous injection.(5) vitamin medicaments is as vitamin C, B, B6 etc.(6) corticosteroid hormone class, it is extensive to be used for skin lesion, and inflammation is rapid, and the person that do not fail of the effect as stated above is as prednisone etc.(7) histamine's globulin is the injection of making after being mixed by hydrochloric acid histamine and gamma globulin, the eczema that is used for showing effect repeatedly.(8) use antimicrobial drug at secondary infection.(9) external therapy (Calamine Lotion etc.).When serious, adopt irradiation: can adopt the superficial X-ray irradiation or treat with radiosiotope phosphorus 32 and strontium 90 to chronic eczema.General exit dose is 80～100 rads, and every day 1 time, 10 times was 1 course of treatment.Total amount generally is no more than 1000 rads, but very plump disease is decreased available to 1500 rads.Subacute eczema has and does not ooze out the also available radioactive isotope therapy of person on a small quantity, its amount of beginning with 30 rads about for well, waiting to ooze out can increase after disappearing gradually, but maximum should not surpass 800 rads, general total amount is 500～800 rads.
The traditional Chinese medical science is advocated giving consideration to both the incidental and fundamental, and the integral body of internal-external therapy and the local principle that combines had both been paid attention to damp and hot performance, paid attention to the basic reason of dysfunction of the spleen in transportation again.On method of treatment, control its mark earlier, treat damp and hot stepping back, then build spleen and help fortune with Zhi Qiben.(1) Coryza Treated by Syndrome Differentiation: acute stage, can be divided into wind heat and contain, and rheumatism is accumulate skin, damp-heat gluing each other (weight in wet base is in heat, and thermogravimetric is in wetting damp and hot laying equal stress on), the wet resistance of stagnation of liver-QI, pattern of syndrome such as spleen-dampness and stomach-heat.Adopt the wind and heat dispersing drink respectively, XIAOFENG SAN, removing damp-heat soup, pathogenic wind expelling daochi powder, removing damp to relieve itching soup, Danzhi Xiaoyao San, the treatment of XIEHUANGSAN plus-minus.(2) other therapies: mainly be Chinese medicine lotion, application, liniment and acupoint injection therapy, umbilicus etc. is applied in cupping.As use acupuncture therapy Changqiang acupoint acupoint injection therapy 5% Radix Angelicae Sinensis injection and promethazine hydrochloride, and can declare venting QI-blood, antagonism histamine suppresses allergy, reduce capillary permeability, improve lymph circulation, blood circulation promoting and blood stasis dispelling, anti-inflammatory analgetic promotes skin lesion to disappear the purpose that reach antipruritic, sepage stops.Meridians are annotated oxygen therapy; Draw pure oxygen 80ml with the 100ml syringe, at place, bladder meridian Geshu cave, back subcutaneous injection, every side 40ml, the next day 1 time, 10 days 1 courses of treatment.This method may be to reach therapeutic purposes by the immune function of regulating human body.
In sum, the pathology factor of relevant eczema is not clear, so doctor trained in Western medicine all belongs to the verification treatment.As use weak curative effects such as antihistaminic, topical application corticosteroid hormone, and the relapse rate height, prolonged application has good action scarcely.Treatment by Chinese herbs determined curative effect, but the effective Chinese patent medicine preparation of clinical shortage.
Still there is not at present the relevant report that separately Radix Scutellariae, Radix Gentianae compatibility is used for the treatment of eczema.
Summary of the invention
Technical scheme of the present invention relates to a kind of pharmaceutical composition for the treatment of eczema.The present invention also provides this preparation of drug combination method and purposes.
The invention provides a kind of pharmaceutical composition for the treatment of eczema, it is the preparation that is prepared from by the following weight proportion raw material:
Radix Scutellariae 1-9 part, Radix Gentianae 9-1 part.
Further preferably, it is the preparation that is prepared from by the following weight proportion raw material:
1 part of Radix Scutellariae, Radix Gentianae 1-2 part.
Still more preferably, it is the medicament that is prepared from by the following weight proportion raw material:
1 part of Radix Scutellariae, 1 part of Radix Gentianae.
Medicine of the present invention is to be active component by the water of Radix Scutellariae, Radix Gentianae or extractive with organic solvent, adds the medicament that acceptable accessories or complementary composition are prepared from.
Wherein, described medicament is: capsule, tablet, pill or granule.
Wherein, in the described capsule every contain baicalin (C
21H
18O
11) should be no less than 35mg, contain gentiopicrin (C
16H
20O
9) should be no less than 12mg.
The present invention also provides this preparation of drug combination method, and it comprises the steps:
A, get it filled material Radix Scutellariae, Radix Gentianae;
B, adding decocting boil, and perhaps adopt the ethanol extraction more than 40%, reclaim ethanol, filter, and drying adds acceptable accessories or complementary composition and is prepared into medicament pharmaceutically commonly used.
Wherein, the described concentration of alcohol of b step is: 60% ethanol.
The present invention also provides the purposes of this pharmaceutical composition in the medicine of preparation treatment eczema.
The present invention also provides the purposes of this pharmaceutical composition in the medicine of preparation treatment acute dermatitis.
Radix Gentianae, Radix Scutellariae compatibility treatment eczema in the medicine material material of the present invention, prove through animal experiment study, Radix Gentianae, the two compatibility of Radix Scutellariae, effect with Synergistic, the damp and hot effectiveness of purging liver and gallbladder heightens, significant antiinflammatory, antiallergic, antipruritic and bacteriostasis are arranged, and it is evident in efficacy to be used for skin pruritus, erythema, pimple or vesicle that acute eczema, dermatitis cause.
The specific embodiment
The preparation of embodiment 1 medicine of the present invention
Radix Scutellariae 667g Radix Gentianae 667g
More than two the flavor raw medicinal materials make 1000 of capsules altogether.
More than two flavors, be ground into coarse powder, with 60% alcohol reflux 2 times, add 12 times of amounts the first time, extracted 2 hours; For the second time add 8 times of amounts, extracted 1 hour, merge alcohol extract, decompression recycling ethanol, being concentrated into relative density is the clear paste of 1.36～1.38 (80 ℃ of heat are surveyed), drying under reduced pressure is pulverized, and crosses 60 mesh sieves, loads capsule, makes 1000, promptly.
Clinical acute eczema and the acute dermatitis of being used for of this product took for 2～4 weeks 3 of every days oral 1 time (0.4g content/grain perhaps is equivalent to 1.33g crude drug in whole/grain) every day 3 times, was the human safe dose and the course of treatment.
Different process screening of the present invention is compared:
Active constituent content is relatively in the different solvents extract:
Take by weighing medicinal material coarse powder 200g in the prescription ratio, totally 4 parts, selecting water, 40%, 60%, 80% ethanol respectively for use is that solvent carries out reflux (or decoction) extraction 3 times, adds 10 times of amounts for the first time, extracts 2 hours; For the second time add 8 times of amounts, extracted 1 hour; Add 7 times of amounts for the third time, extracted 1 hour.Merge extractive liquid,, standardize solution is as need testing solution.The accurate need testing solution 10 μ l that draw handle by gentiopicroside in different morphological assay method under baicalin, the Radix Gentianae item under Radix Scutellariae item of Chinese Pharmacopoeia version in 2000, and parallel assay 2 times the results are shown in Table 1.
Mainly contain effective component content (g) in the different extracting solution of table 1
| Extract solvent | Water | 40% ethanol | 60% ethanol | 80% ethanol |
| Content of baicalin | 8.19 | 8.59 | 8.87 | 4.82 |
| Gentiopicroside in different morphological | 3.82 | 3.71 | 3.80 | 3.93 |
As seen from Table 1, different extraction solvents are little to the influence of gentiopicrin yield, and very big to the influence of baicalin yield, the optimum extraction solvent is 60% ethanol, is water, 40% ethanol secondly.
The preparation of embodiment 2 medicines of the present invention
Radix Scutellariae 667g Radix Gentianae 74g
More than two the flavor raw medicinal materials make 1000 of capsules altogether.
More than two flavors, be ground into coarse powder, with 80% alcohol reflux 2 times, add 12 times of amounts the first time, extracted 2 hours; For the second time add 8 times of amounts, extracted 1 hour, merge alcohol extract, decompression recycling ethanol, being concentrated into relative density is the clear paste of 1.36～1.38 (80 ℃ of heat are surveyed), drying under reduced pressure is pulverized, and crosses 60 mesh sieves, loads capsule, makes 1000, promptly.
The preparation of embodiment 3 medicines of the present invention
Radix Scutellariae 74g Radix Gentianae 667g
More than two the flavor raw medicinal materials make 1000 in tablet altogether.
More than two flavors, be ground into coarse powder, with 95% alcohol reflux 2 times, add 12 times of amounts the first time, extracted 2 hours; For the second time add 8 times of amounts, extracted 1 hour, merge alcohol extract, decompression recycling ethanol, being concentrated into relative density is the clear paste of 1.36～1.38 (80 ℃ of heat are surveyed), and drying under reduced pressure is pulverized, and crosses 60 mesh sieves, adds magnesium stearate, and tabletting is made 1000, promptly.
The preparation of embodiment 4 medicines of the present invention
Radix Scutellariae 667g Radix Gentianae 1334g
More than two the flavor raw medicinal materials make 1000 in tablet altogether.
More than two flavors, be ground into coarse powder, with 40% alcohol reflux 2 times, add 12 times of amounts the first time, extracted 2 hours; For the second time add 8 times of amounts, extracted 1 hour, merge alcohol extract, decompression recycling ethanol, being concentrated into relative density is the clear paste of 1.36～1.38 (80 ℃ of heat are surveyed), and drying under reduced pressure is pulverized, and crosses 60 mesh sieves, adds magnesium stearate, and tabletting is made 1000, promptly.
Embodiment 5 drug quality control methods of the present invention
With baicalin, gentiopicroside in different morphological is index, has set up high-performance liquid chromatogram determination method, by the assay to three batches of pilot products, has worked out its content limit (width of cloth) degree index and assay method:
Baicalin and gentiopicroside in different morphological HPLC assay method are as follows:
Baicalin is measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000).
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-water-phosphoric acid (47: 53: 0.2) is a mobile phase; The detection wavelength is 280nm.Theoretical cam curve is calculated with the baicalin peak should be not less than 2500.
The preparation precision of reference substance solution takes by weighing at 4 hours baicalin reference substance of 60 ℃ of drying under reduced pressure an amount of, adds methanol and makes the solution that every 1ml contains 60 μ g, promptly.
It is an amount of that the orthogonal test sample is got in the preparation of need testing solution, and accurate the title decides, and puts in the 25ml volumetric flask, uses the mobile phase ultrasonic dissolution, shakes up, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
Gentiopicrin is measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-water (11: 89) is a mobile phase; The detection wavelength is 270nm.Theoretical cam curve is calculated with the gentiopicrin peak should be not less than 4000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the gentiopicrin reference substance, adds methanol and make the solution that every 1ml contains 0.3mg, promptly.
It is an amount of that the orthogonal test sample is got in the preparation of test solution, the accurate title, decide, put in the 10ml volumetric flask, the accurate methanol 10ml that adds shakes up, and ultrasonic (power is not less than 100W, frequency is not less than 40kHz) handled 15 minutes, leave standstill, centrifugal (Revolution Per Minute 3000 changes), supernatant is need testing solution.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
Below prove beneficial effect of the present invention by pharmacodynamics test.
1, is subjected to reagent thing and positive drug
Medicament capsule of the present invention: embodiment 1 preparation.Specification 0.4g (content)/grain.Heat clearing away, dampness, antipruritic.Be used for skin pruritus, erythema, pimple or vesicle that acute eczema, dermatitis cause.Its content is pale brown toner end, and 1g medicated powder is equivalent to crude drug in whole 3.34g, and lot number 20030508 is provided by medicine chamber, Traditional Chinese Medicine Research Institute, Sichuan Province.
Sample 1 (KM-B): pale brown toner end, 1g medicated powder is equivalent to crude drug in whole 3.31g, and lot number 20030405 is provided by medicine chamber, Traditional Chinese Medicine Research Institute, Sichuan Province Yang Andong assistant researcher.
The preparation method of sample 1: take by weighing 50 times of medicine material of the present invention (Radix Scutellariae 667g Radix Gentianae 667g),, add 10 times of amounts for the first time, extracted 2 hours with water extraction three times; For the second time add 8 times of amounts, extracted 1 hour; Add 7 times of amounts for the third time, extracted 1 hour, filter, merging filtrate is condensed into clear paste, the clear paste drying under reduced pressure, and yield is 33.0% (numbering KM-B).
Sample 2 (KM-A): pale brown toner end, 1g medicated powder is equivalent to crude drug in whole 3.03g, and lot number 20030406 is provided by medicine chamber, Traditional Chinese Medicine Research Institute, Sichuan Province Yang Andong assistant researcher.
The preparation method of sample 2: take by weighing 50 times of medicine material of the present invention (Radix Scutellariae 667g Radix Gentianae 667g),, add 10 times of amounts for the first time, extracted 2 hours with 60% ethanol extraction three times; For the second time add 8 times of amounts, extracted 1 hour; Add 7 times of amounts for the third time, extracted 1 hour, filter merging filtrate.Decompression recycling ethanol gets clear paste, and clear paste subtracts drying, and yield is 30.2% (numbering KM-A).
Ibuprofen (ibuprofen modified release capsule): (90) are defended the accurate word X-129 of medicine number.Specification: ibuprofen 300mg/ grain, Sino-U.S. Tianjin SmithKline Pharmacy stock Co., Ltd produces, lot number 20021120.
Dexamethasone acetate tablets: the accurate word (1996) of medicine is defended No. 012641 in the river.Specification 0.75mg * 100 slice/bottle, Mt. Tiantai, Chengdu pharmaceutical Co. Ltd produces, lot number 980401.
Benorilate tablet (benorilate tablet): the accurate word (1981) of medicine is defended No. 000144 in the river.Specification 0.5g * 100 slice, Chengdu pharmaceutcal corporation, Ltd produces, lot number 990926.
Chlorpheniramine maleate tablets: the accurate word (1993) of medicine is defended No. 008690 in the river.Specification 4mg * 100 slice/bottle, Leshan, Sichuan the 3rd pharmaceutical factory produces, lot number 20020101.
2, animal
Km (Kunming kind) mice, one-level is qualified, and the real kinoplaszm in river 2002-33 number is provided by the Traditional Chinese Medicine Research Institute, Sichuan Province Experimental Animal Center.
The SD rat, one-level is qualified, and the real kinoplaszm in river 2002-32 number is provided by the Traditional Chinese Medicine Research Institute, Sichuan Province Experimental Animal Center.
3, instrument and reagent
3.1 instrument
Day island proper Tianjin UV-730 automatic biochemical analyzer.
Day island proper Tianjin EB-3200D precise electronic balance (precision 0.01g).
The JA1003A of Shanghai Jingtian Electronic Instrument Co., Ltd. type electronic balance (precision 1mg).
Rat foot volume determination instrument.
The 80-2 of Shanghai Surgical Operation Equipment Factory centrifugation device.
3.2 reagent
Dimethylbenzene: the 500ml/ bottle, analytical pure, chemical reagent factory in Chengdu produces, lot number 20021120.
Carrageenin: specification 10g/ bottle, Shenyang Pharmacy College provides, lot number 980801.Be mixed with 1% solution with distilled water the previous day in test, put 4 ℃ of refrigerators and preserve standby.
Azovan blue: the 10g/ bottle, Fluka import packing, Shanghai chemical reagent purchasing and supply station is sold, lot number 98-11-02.
Acetone: 500ml/ bottle, analytical pure, the Long Huagongshijichang of Chengdu section, lot number 20030110.
Sodium sulfide: 500g/ bottle, chemical pure, Chengdu chemical reagent factory, lot number 980923.
Ovalbumin: the 50g/ bottle is provided lot number Lot#0123 by sigma company.
Absorption whooping cough, diphtheria, tetanus combined vaccine: the 5ml/ ampoule, 10/box, defend system (82) Rong (4) No. 01, lot number 20020403 is provided by Chengdu Inst. of Biological Products.
Ether: the 500ml/ bottle, analytical pure, rich Dihua, Tianjin worker's company limited is produced lot number 20020208.
2,4-dinitrochlorobenzene (DNCB): 25g/ bottle, chemical pure, Shanghai chemical reagent one factory, lot number 890901.
Sodium dihydrogen phosphate (NaH
2PO
4.2H
2O): 500g/ bottle, analytical pure, Shantou City's brilliance laboratory, lot number 20010811.
Sodium hydrogen phosphate (Na
2HPO
4.12H
2O): 500g/ bottle, analytical pure, Shantou City's brilliance laboratory, lot number 20010820.
Histamine phosphate: 5g/ bottle, content＞98.0%, Chinese Academy of Sciences Shanghai east wind Biochem Technology, INC. of biochemical institute, lot number 9611147.
The Dextran 40 glucose injection: 500ml/ bottle, river are defended the accurate word (1994) of medicine No. 010577, Leshan, Sichuan second pharmaceutical factory, lot number 0006147-5.
0.9% sodium chloride injection: 500ml/ bottle, river are defended the accurate word (1996) of medicine No. 012141, big pharmaceutical factory, Cologne, Sichuan, lot number 021020-14.
4, animal
Km (Kunming kind) mice, one-level is qualified, and the real kinoplaszm in river 2002-33 number is provided by the Traditional Chinese Medicine Research Institute, Sichuan Province Experimental Animal Center.
The different proportion compatibility combinations of test example 1 medicine of the present invention are to the influence of mouse skin contact delayed hypersensitivity (DTH)
100 of Kunming mouses, female, body weight 18～22g is divided into 10 groups at random by body weight, 10 every group.If normal control (blank), model contrast (solvent control) and positive controls (dexamethasone acetate tablets), four to ten groups is the different proportion compatibilities combinations of medicine of the present invention, presses the preparation of embodiment 1 method, dosage 4g (crude drug in whole)/kg.Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, continuous 13 days.5h before the first administration is with 8%Na
2S loses hair or feathers each group mouse back skin in advance, except that the normal control group, all the other each groups all only are coated with 10%DNCB acetone solution 20 μ l/ in back depilation place, 2h after the administration in the 12nd day, each is organized mice and all only evenly is applied to the auris dextra tow sides with 1%DNCB acetone solution 15 μ l/, behind the 24h, be that the last administration is after 50 minutes, disconnected neck is put to death mice, cuts two ears along the auricle baseline, represents its anaphylaxis degree with the swelling degree (mg) of the difference of two auricle weight.The result relatively respectively organizes with the t check and asks the swelling degree significance of difference, inspection level α=0.05.
By following table as seen, (Radix Scutellariae: Radix Gentianae=9: 1～1: 9) all mouse skin contact delayed hypersensitivity is had inhibitory action (p＜0.05～0.01) or inhibitory action trend (p＞0.05), suppression ratio is between 17.14～56.19% for the different proportion compatibility combinations of medicine of the present invention.Radix Scutellariae wherein: Radix Gentianae is 1: effect is more excellent in the time of (1-2), and with Radix Scutellariae: when Radix Gentianae was 1: 1 group, inhibitory action was the strongest, was best proportion compatibility group.
The different proportion compatibility combinations of table 2 medicine of the present invention are to the influence of mouse skin contact delayed hypersensitivity
| Group | Number of animals (only) | Dosage (g/kg * d) | Anaphylaxis degree (mg) | Anaphylaxis suppression ratio (%) |
| Normal control | 10 | - | 3.29±2.43 *** | - |
| Model contrast dexamethasone Radix Scutellariae 9: Radix Gentianae 1 Radix Scutellariae 5: Radix Gentianae 1 Radix Scutellariae 2: Radix Gentianae 1 Radix Scutellariae 1: Radix Gentianae 1 Radix Scutellariae 1: Radix Gentianae 2 Radix Scutellariaes 1: Radix Gentianae 5 Radix Scutellariaes 1: Radix Gentianae 9 | 10 10 10 10 10 10 10 10 10 | - 1mg/kg×13 4.0×13 4.0×13 4.0×13 4.0×13 4.0×13 4.0×13 4.0×13 | 10.32±2.37 4.27±2.29 *** 6.88±2.54 *** 7.21±3.23 ** 6.83±3.44 ** 4.98±2.91 *** 5.97±3.21 ** 6.83±2.27 ** 6.55±3.54 ** | - 58.62 33.33 30.14 33.82 51.74 42.15 33.82 36.53 |
Compare with model group:
*P＞0.05
*P＜0.05;
* *P＜0.01.
After having determined the best compatibility situation of raw material, below be that the different extraction processes of raw material are tested.
The different extraction processes of test example 2 medicine materials of the present invention are to the influence of mouse dimethylbenzene ear expanding
1, principle: smear certain density dimethylbenzene for the mice auricular concha and can cause and bring out mice auricular concha acute inflammation edema.This model can be observed medicine different process sample of the present invention to the early stage antagonism of inflammation.
2, method:
The skullcapflavone compounds has tangible anti-allergy action, and aglycon is strong than the glycosides effect, extracts solvent and preferably selects ethanol for use.Consider to have the hydrotropy effect between the composition, also the possibility water is fried for aglycon.For the selective extraction solvent, water, 60% ethanol are for extracting solvent respectively, and the preparation sample carries out efficacy trial, and the result shows that alcohol extraction is better than water and carries.
Test specimen: sample thief 1 (numbering KM-B), sample 2 (numbering KM-A) are tested.
60 of Kunming mouses, male, body weight 18～22g is divided into 6 groups at random by body weight, 10 every group.If negative control (solvent control) and positive controls (ibuprofen capsule), be subjected to KM-A2 dosage group of reagent thing (be equivalent to 2 respectively, 4g crude drug in whole/kg), be subjected to KM-B2 dosage group of reagent thing (be equivalent to 2 respectively, 4g crude drug in whole/kg).Adopt not isoconcentration medicinal liquid gastric infusion (matched group is given distilled water) of isometric(al), be subjected to that KM-A2 dosage group of reagent thing concentration is respectively 20,40g crude drug in whole/dl, be subjected to that KM-B2 dosage group of reagent thing concentration is respectively 20,40g crude drug in whole/dl, ibuprofen concentration is 2g/dl, irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, for three days on end.40min after the last administration, only evenly being applied in the mouse right ear tow sides causes inflammation (except the matched group) with dimethylbenzene 0.03ml/, puts to death animal after 20 minutes, cuts two ears along the auricle baseline,, and calculate and respectively organize inhibitory rate of intumesce (%) as swelling degree (mg) with the difference of two auricle weight.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
The different extraction process sample of table 3 medicament capsule of the present invention xylol brings out the influence of chmice acute ear swelling
Compare (t check or t ' check) with matched group:
*P＞0.05;
*P＜0.05;
* *P＜0.01.
3, result: as seen by table 2, sample KM-A2 dosage group and be subjected to KM-B2 dosage group of reagent thing xylol to cause the chmice acute ear swelling remarkable inhibitory action is all arranged, wherein the acutely inflamed suppression ratio of KM-A xylol and a little more than KM-B has certain difference trend.
The different extraction processes of test example 3 medicines of the present invention are to the influence of mouse skin contact delayed hypersensitivity (DTH)
1, principle: behind the hapten contact mouse web portion epidermis (sensitization),, combine, and antigenic peptides is passs the dermatogen specific T-cells with Langhans' cells MHC II class antigen through chemical modification.After the sensitization 6 days, Mice Auricle contacted hapten (triggering) once more, and activation CD4+T cell produces the various kinds of cell factor, triggers contact hypersensitivity, Mice Auricle edema (people: allergic contact dermatitis).This model can be observed the antagonism of medicine different process sample of the present invention to delayed hypersensitivity.
2, method: 77 of Kunming mouses, female, body weight 18～22g is divided into 7 groups at random by body weight, 11 every group.If normal control (blank), model contrast (solvent control) and positive controls (dexamethasone acetate tablets), be subjected to 2 dosage groups of reagent thing KM-A (be equivalent to 2 respectively, 4g crude drug in whole/kg), be subjected to 2 dosage groups of reagent thing KM-B (be equivalent to 2 respectively, 4g crude drug in whole/kg).Adopt not isoconcentration medicinal liquid gastric infusion (matched group is given distilled water) of isometric(al), be subjected to that KM-A2 dosage group of reagent thing concentration is respectively 20,40g crude drug in whole/dl, be subjected to that KM-B2 dosage group of reagent thing concentration is respectively 20,40g crude drug in whole/dl, dexamethasone concentration is 0.01g/dl, irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, continuous 13 days.5h before the first administration is with 8%Na
2S loses hair or feathers each group mouse back skin in advance, except that the normal control group, all the other each groups all only are coated with 10%DNCB acetone solution 20 μ l/ in back depilation place, 2h after the administration in the 12nd day, each is organized mice and all only evenly is applied to the auris dextra tow sides with 1%DNCB acetone solution 15 μ l/, behind the 24h, be that the last administration is after 50 minutes, disconnected neck is put to death mice, cuts two ears along the auricle baseline, represents its anaphylaxis degree with the swelling degree (mg) of the difference of two auricle weight.Cut open simultaneously and get spleen and thymus, weigh, calculate organ coefficient.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
5.2.3 result: as seen by table 3, sample KM-A 2,4g (crude drug in whole)/2 dosage groups of kg and be subjected to the comparison of reagent thing KM-B 4g (crude drug in whole)/kg dosage group and model group, its anaphylaxis degree reduces significantly that (significant difference is remarkable, P＜0.05 or 0.01), is subjected to the visible certain effect trend of reagent thing KM-B 2g (crude drug in whole)/kg dosage group.The different extraction process sample KM-A and the KM-B that show drug sample of the present invention all have inhibitory action to DNCB induced mice contact skin delayed skin hypersensitivity, and the anti-allergic effects of KM-A is better than the anti-allergic effects of KM-B under same dose.In addition, observe also that significantly to suppress immune organ different with dexamethasone, this product does not have obvious influence to the organ coefficient of immune organ spleen, thymus in effective dosage ranges.
The different extraction process samples of table 4 medicine of the present invention are to the influence of mouse skin contact delayed hypersensitivity (PTH)
| Group | Number of animals (only) | Dosage (g crude drug in whole/kg * d) | Anaphylaxis degree (mg) | Anaphylaxis suppression ratio (%) | Thymus organ index (mg/10g) | Spleen organ index (mg/10g) |
| Normal control model contrast KM-AKM-AKM-BKM-B dexamethasone | 11 11 11 11 11 11 11 | - - 2×13 4×13 2×13 4×13 1mg/kg×13 | 4.42±2.64 ***9.36±3.935.82±2.52 **5.00±2.83 ***6.46±4.50 *5.09±3.56 **4.82±2.89 *** | - - 37.82 46.58 30.98 45.61 48.50 | 45.00±8.64 *49.47±13.3344.83±5.64 *46.91±10.09 *44.74±6.84 *51.86±11.42 *12.25±4.10 *** | 53.14±19.59 *54.81±26.4264.10±35.31 *58.86±23.75 *64.84±32.09 *46.80±9.97 *23.48±12.48 *** |
Compare (t check) with model group: * P＞0.05; * P＜0.05; * * P＜0.01.
3, conclusion (of pressure testing)
3.1 a medicine KM-A2 of the present invention dosage group and KM-B2 dosage group xylol cause the chmice acute ear swelling remarkable inhibitory action is arranged all, wherein the acutely inflamed suppression ratio of KM-A xylol and a little more than KM-B has certain difference trend.
3.2 medicine KM-A2 of the present invention, 4g (crude drug in whole)/2 dosage groups of kg and KM-B 4g (crude drug in whole)/kg dosage group and model group are relatively, its anaphylaxis degree reduces significantly that (significant difference is remarkable, P＜0.05 or 0.01), is subjected to the visible certain effect trend of reagent thing KM-B 2g (crude drug in whole)/kg dosage group.Show that different extraction process sample KM-A of medicine of the present invention and KM-B all have inhibitory action to DNCB induced mice contact skin delayed skin hypersensitivity, the anti-allergic effects of KM-A is better than the anti-allergic effects of KM-B under same dose.In addition, observe also that significantly to suppress immune organ different with dexamethasone, in effective dosage ranges, the organ coefficient of immune organ spleen, thymus is not had obvious influence.
3.3 from institute's amount of reagent scope and this experimental model index result as can be seen, medicine KM-A of the present invention and KM-B all have anti-inflammation detumescence and anti-allergic effects, wherein, the anti-inflammation detumescence of medicine KM-A of the present invention and anti-allergic effects are better than medicine KM-B of the present invention.
In sum, medicine material alcohol extraction effect of the present invention is better than water to be carried, and extracting method selects 60% ethanol to carry out reflux, extract, for extracting solvent.
Test example 4 medicine resist inflammation on repercussive function of the present invention
Often with inflammatory reaction, the Chinese medicine of great majority treatment eczema all has the effect of certain inflammation-inhibiting reaction to eczema in pathogenic process.This product is mainly used in acute eczema, acute dermatitis, so based on the acute inflammation model test.
1, to the influence of mouse dimethylbenzene ear expanding
(1) purpose: smear certain density dimethylbenzene for the mice auricular concha and can cause and bring out mice auricular concha acute inflammation edema.This model can be observed medicament capsule of the present invention to the early stage antagonism of inflammation.
(2) method: 55 of Kunming mouses, male, body weight 18～22g is divided into 5 groups at random by body weight, 11 every group.If negative control (solvent control) and positive controls (ibuprofen capsule) are subjected to 3 dosage groups of reagent thing (1,2,4g crude drug in whole/kg, be equivalent to 5,10,20 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion (matched group is given distilled water) of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 10,20,40g crude drug in whole/dl, ibuprofen concentration is 2g/dl, and irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, for three days on end.40min after the last administration, only evenly being applied in the mouse right ear tow sides causes inflammation (except the matched group) with dimethylbenzene 0.03ml/, puts to death animal after 20 minutes, cuts two ears along the auricle baseline,, and calculate and respectively organize inhibitory rate of intumesce (%) as swelling degree (mg) with the difference of two auricle weight.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
(3) result: by table 5 as seen, the interior xylol of medicament capsule institute's amount of reagent scope of the present invention causes the chmice acute ear swelling remarkable inhibitory action, and the different trend of a certain amount of heterodyne is arranged, and shows that the acute inflammation that medicament capsule xylol of the present invention causes has antagonism.
Table 5 medicament capsule xylol of the present invention brings out the influence of chmice acute ear swelling
Compare (t check or t ' check) with matched group: * P＞0.05; * P＜0.05; * * P＜0.01.
2, on Carrageenan (carrageenin) is brought out the influence of rat foot claw swelling
(1) purpose: be chosen in the effect proinflammatory agent carrageenin local injection that injects 2～4 hours swelling peakings in back and bring out rat foot claw swelling, the characteristics of this model are to cause the synthetic increase of scorching local PG, and bring out edema with vaso-active substance and kassinin kinin class.Cause the antiinflammatory action that medicament capsule of the present invention is observed in the volumetrical variation of scorching front and back rat paw edema by measurement.
(2) method: 50 of SD rats, male, body weight 100～140g is divided into 5 groups at random by body weight, 10 every group.If negative control (solvent control) and positive controls (benorilate tablet) are subjected to 3 dosage groups of reagent thing (0.8,1.6,3.2g crude drug in whole/kg, be equivalent to 4,8,16 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 8,16,32g crude drug in whole/dl, benorylate concentration is 2g/dl, irritates stomach volume 1ml/100g body weight (matched group is given the isometric(al) distilled water), every day 1 time, for three days on end.40min injects 1% carrageenin 0.05ml down for the right back sufficient plantar aponeurosis of rat and causes inflammation after the last administration.Adopt rat foot claw volume determination instrument to measure to cause scorching before and cause scorching back different time (at interval 1～2 hour once, continuous 5 times) rat foot claw volume-variation, be calculated as follows the rat paw edema rate.Calculate and respectively organize swelling rate (%), and do the t check, relatively each group is asked the significance of difference, inspection level α=0.05.
(3) result: as seen by table 6, medicament capsule 1.6 of the present invention, 3.2g (crude drug in whole)/kg dosage group swelling rate are lower than matched group swelling rate, learn by statistics to handle and have the significance of difference, the different trend of a certain amount of heterodyne is arranged, mainly act on behind the carrageenin local injection sufficient pawl swelling in 1 hour and form phase and injection back swelling in 4-8 hour paracmasis, show that medicament capsule on Carrageenan local injection of the present invention brings out rat foot claw swelling and has obvious inhibitory action.
The inhibitory action that table 6 medicament capsule on Carrageenan of the present invention is brought out rat foot claw swelling
Compare (t check) with matched group: * P＞0.05; * P＜0.05; * * P＜0.01.
3, to the influence of mouse skin capillary permeability
(1) purpose: select dimethylbenzene to make pro-inflammatory cytokine, local excitation mouse skin capillary permeability increases.Vein injects azovan blue, and this dyestuff can go out to vascular exosmosis in hyperfunction blood capillary, and how much can be used as of dyestuff seepage discharge judges whether medicament capsule of the present invention can the hyperfunction index of the early stage capillary permeability of inflammation-inhibiting.
(2) method: 60 of Kunming mouses, male and female half and half, body weight 18～21g is divided into 5 groups at random by body weight, 12 every group.If negative control (solvent control) and positive controls (dexamethasone acetate tablets) are subjected to 3 dosage groups of reagent thing (1,2,4g crude drug in whole/kg, be equivalent to 5,10,20 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 10,20,40g crude drug in whole/dl, dexamethasone acetate tablets concentration is 45mg/dl, and irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, for three days on end.Melted paraxylene 0.02ml/ was only on the skin of mouse web portion centre position immediately through mouse tail vein injection 0.5% azovan blue normal saline 0.1ml/10g body weight in 1 hour after the last administration.Animal is put to death in the cervical vertebra displacement behind the 20min, cuts the skin of locus coeruleus, shreds.Be soaked in the having in the plug glass centrifuge tube of acetone-sodium sulfate liquid is housed, be settled to 10ml, lucifuge was deposited 3 days, shook this test tube every day gently 2～3 times, and centrifugal 10 minutes of 2000pm gets supernatant at 590nm place mensuration absorbance (A value).The result compares each group difference significance, inspection level α=0.05 with the t check.
(3) result: by table 7 as seen, medicament capsule of the present invention tries 3 dosage group locus coeruleus extraction dyestuff absorbances and significantly is lower than matched group, shows that medicament capsule of the present invention can increase the mouse skin capillary permeability that anti-inflammatory causes.
Table 7 medicament capsule xylol of the present invention brings out the influence that the mouse skin capillary permeability increases
Compare (t check) with matched group: * P＞0.05; * P＜0.05; * * P＜0.01.
Test example 5 medicine anti-allergic effects of the present invention
Think that at present eczema is a kind of allergic disease, the anti-allergic effects of research Chinese medicine should be the major part of the pharmacodynamic study of its treatment eczema.
1, medicine of the present invention is to the influence of rat passive cutaneous anaphylaxis, PCA of the same race (Rat PCA)
(1) purpose: intradermal injection is in normal rat stomach wall or back with the serum (including abundant Ig E antibody) of sensitization rat, and the Fc receptors bind of Ig E and local skin mastocyte makes it passive allergy.When antigen is attacked, cause that local mastocyte discharged sensitive media, thereby the permeability of local vascular is increased, judge that with ivens is blue vascular permeability changes, and can reflect the skin allergy degree.
(2) method
Antiserum Preparation: get 10 of rats, hind leg both sides intramuscular injection egg protein normal saline solution 30mg/kg.2ml
-1, lumbar injection PertussisDiphtheriaTetanus triple vaccine 2.5ml/ only simultaneously.Duplicate injection 1 time again in the 2nd day was put to death animal after 14 days, the femoral artery blood-letting, and 4 ℃ of 2500rpm * 10min separation of serum ,-30 ℃ of preservations are standby.
Attack and administration: get 50 of SD rats, male and female half and half, body weight 150～200g is divided into 5 groups at random by body weight, 10 every group.If negative control (solvent control) and positive controls (chlorpheniramine maleate tablets) are subjected to 3 dosage groups of reagent thing (0.8,1.6,3.2g crude drug in whole/kg, be equivalent to 4,8,16 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 8,16,32g crude drug in whole/dl, chlorphenamine concentration is 0.032g/dl, irritates stomach volume 1ml/100g body weight (matched group is given the isometric(al) distilled water), every day 1 time, continuous 5 days.Therebetween, after 3 days, each treated animal is all in back spinal column both sides cropping in administration, and every Mus is cut 2 points, about 1cm * 1cm size.Left side injection dilution in 1: 2 antiserum, right side injection dilution in 1: 4 antiserum, 0.1ml/ point.Injection back 48h, each caudal vein is only injected blue physiological salt liquid (the including the 2mg/ml egg protein) 2ml/ of 1% ivens, disconnected neck is put to death animal behind the 20min, cut indigo plant and dye skin, be soaked in 4ml acetone-metabisulfite solution, the centrifugal 10min of 2000rpm after 3 days gets supernatant in 590nm place colorimetric determination optical density, and is zero with the acetone-sodium sulfate liquid that does not steep skin.Blue mean and the standard deviation of dying skin soak OD value of each group of calculating, and do the t check, compare each group difference significance, inspection level α=0.05.
Table 8 medicament capsule of the present invention is to the influence of rat passive cutaneous anaphylaxis, PCA of the same race (Rat PCA)
Compare (t check) with matched group: * P＞0.05; * P＜0.05; * * P＜0.01.
(3) result: as seen by table 8, medicament capsule 1.6 of the present invention, 3.2g/kg skin locus coeruleus soak OD value are lower than matched group (P＜0.05～0.01), 0.8g/kg visible certain trend (P＞0.05) shows that medicament capsule of the present invention is inhibited to rat passive cutaneous anaphylaxis, PCA of the same race.
2, medicine of the present invention is to the influence of mouse skin contact delayed hypersensitivity (PTH)
(1) purpose: behind the hapten contact mouse web portion epidermis (sensitization),, combine, and antigenic peptides is passs the dermatogen specific T-cells with Langhans' cells MHC II class antigen through chemical modification.After the sensitization 6 days, Mice Auricle contacted hapten (triggering) once more, and activation CD4+T cell produces the various kinds of cell factor, triggers contact hypersensitivity, Mice Auricle edema (people: allergic contact dermatitis).
(2) method: 60 of Kunming mouses, female, body weight 18～22g is divided into 6 groups at random by body weight, 10 every group.If negative control (solvent control) and positive controls (dexamethasone acetate tablets) are subjected to 3 dosage groups of reagent thing (1,2,4g crude drug in whole/kg, be equivalent to 5,10,20 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 10.0,20.0,40.0g crude drug in whole/dl, dexamethasone concentration is 0.01g/dl, and irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, continuous 13 days.5h before the first administration is with 8%Na
2S loses hair or feathers each group mouse back skin in advance, except that the normal control group, all the other each groups all only are coated with 10%DNCB acetone solution 20 μ l/ in back depilation place, 2h after the administration in the 12nd day, each is organized mice and all only evenly is applied to the auris dextra tow sides with 1%DNCB acetone solution 15 μ l/, behind the 24h, be that the last administration is after 50 minutes, disconnected neck is put to death mice, cuts two ears along the auricle baseline, represents its anaphylaxis degree with the swelling degree (mg) of the difference of two auricle weight.Cut open simultaneously and get spleen and thymus, weigh, calculate organ coefficient.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
(3) result: by table 9 as seen, medicament capsule of the present invention tries 3 dosage groups and model group compares, and its anaphylaxis degree significantly reduces (significant difference is remarkable, P＜0.05 or 0.01), and is certain dose-effect relationship.Different with dexamethasone, medicine of the present invention does not have obvious influence to the organ coefficient of immune organ spleen, thymus in effective dosage ranges.Show that medicament capsule of the present invention has inhibitory action to DNCB induced mice contact skin delayed skin hypersensitivity.
Table 9 medicament capsule of the present invention is to the influence of mouse skin contact delayed hypersensitivity (PTH)
| Group | Number of animals (only) | Dosage (g crude drug in whole/kg * d) | Anaphylaxis degree (mg) | Splenic organs index (mg/10g) | Thymus organ index (mg/10g) |
| Matched group model group medicine of the present invention medicine of the present invention medicine dexamethasone of the present invention | 10 10 10 10 10 10 | - - 1×13 2×13 4×13 1mg/kg×13 | 4.00±2.75 *** 1.70±5.74 6.10±5.49 ** 5.00±2.40 *** 4.10±3.18 *** 3.70±2.31 *** | 69.80±18.63 * 61.49±11.93 65.28±30.07 * 62.77±12.39 * 70.19±30.60 * 22.57±2.91 *** | 43.92±12.89 * 52.23±9.46 62.77±49.03 * 44.58±6.36 * 40.74±7.42 * 14.39±8.98 *** |
Compare (t check) with model group: * P＞0.05; * P＜0.05; * * P＜0.01.
3, medicine of the present invention is to the influence of mice intradermal injection delayed hypersensitivity (PTH)
(1) purpose: identical with contact hypersensitivity, when DTH also comprises sensitization and triggers two mutually.Mouse web portion subcutaneous injection hapten (sensitization), antigen atopy T cell distribution is to each lymphoid tissue of periphery.After the sensitization 6 days, mouse insole was injected hapten (triggering) once more, and activation CD4+T cell produces the various kinds of cell factor, triggers DTH (mouse insole swelling).
(2) method: 60 of Kunming mouses, female, body weight 18～22g is divided into 6 groups at random by body weight, 10 every group.If negative control (solvent control) and positive controls (dexamethasone acetate tablets) are subjected to 3 dosage groups of reagent thing (1,2,4g crude drug in whole/kg, be equivalent to 5,10,20 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 10.0,20.0,40.0g crude drug in whole/dl, dexamethasone concentration is 0.01g/dl, and irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, continuous 9 days.3h before the first administration, with 0.03ml/ back intradermal injection of 7%DNCB acetone solution (3: 7) (except the normal group), 2h after the administration in the 9th day, each is organized mice and all fills up injection 0.5%DNCB PBS liquid (PH=7.4) 0.03ml/ fully in the right side only, behind the 24h, disconnected neck is put to death mice, cuts left and right two foots along ankle joint suprapubic hair margin place, represents its anaphylaxis degree with the swelling degree (mg) of the difference of two heavy sensation in the foot amounts.Cut open simultaneously and get spleen and thymus, weigh, calculate organ coefficient.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
Table 10 medicament capsule of the present invention is to the influence of mice intradermal injection delayed hypersensitivity (PTH)
| Group | Number of animals (only) | Dosage (g crude drug in whole/kg * d) | Anaphylaxis degree (mg) | Splenic organs index (mg/10g) | Thymus organ index (mg/10g) |
| Matched group model group medicine of the present invention medicine of the present invention medicine dexamethasone of the present invention | 10 10 10 10 10 10 | - - 1×9 2×9 4×9 1mg/kg×9 | 10.50±5.48 ** 23.90±5.38 20.90±14.64 * 17.00±7.33 ** 12.90±7.72 *** 16.90±8.66 ** | 63.82±10.91 * 69.16±9.98 85.58±44.10 * 73.95±18.63 * 58.84±10.24 * 22.97±3.01 *** | 37.37±13.24 * 44.30±8.56 40.36±10.93 * 45.77±13.19 * 38.09±9.09 * 8.59±3.36 *** |
Compare (t check) with model group: * P＞0.05; * P＜0.05; * * P＜0.01.
(3) result: as seen by table 10, medicament capsule of the present invention tries 2,4g crude drug in whole/kg dosage group and model group relatively, its anaphylaxis degree significantly reduces (P＜0.05 or 0.01), and 1g that medicament capsule of the present invention tries (crude drug in whole)/kg group also has certain effect trend, is certain dose-effect relationship.Different with dexamethasone, this product does not have obvious influence to the organ coefficient of immune organ spleen, thymus in effective dosage ranges.Show that medicament capsule of the present invention has better inhibitory action to mice intradermal injection delayed hypersensitivity.
Test example 6 medicine itching-relieving actions of the present invention
Eczema when morbidity often with violent pruritus, so itching-relieving action should be indispensable in the research process of a treatment by Chinese herbs eczema pharmacodynamics content.
1, medicine of the present invention causes the influence of itching to Cavia porcellus histamine part
(1) purpose: smear a certain amount of variable concentrations histamine and can cause that in the Cavia porcellus local skin its local skin pruritus causes Cavia porcellus to produce the pruritus action, reflects the itching-relieving action of medicine with itch-threshold.
(2) method: get 60 of healthy guinea pigs, male and female half and half, body weight 250 ± 50g is divided into 6 groups at random by body weight, 10 every group.If contrast of normal control, model and positive controls (chlorpheniramine maleate tablets) are subjected to 3 dosage groups of reagent thing (0.8,1.6,3.2g crude drug in whole/kg, be equivalent to 4,8,16 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 8,16,32g crude drug in whole/dl, chlorphenamine concentration is 0.016g/dl, irritates stomach volume 1m1/100g body weight (matched group is given the isometric(al) distilled water), every day 1 time, continuous 4 days.The experiment proxima luce (prox. luc) shaves hair for the right back instep of each Cavia porcellus with razor, and 1h after the last administration will shave hair place skin and abrade right back instep with coarse sandpaper and shave hair place, area 1cm
2, beginning is only dripped 0.01% 0.05ml/ of histamine phosphate at the wound surface place, after this every 3 minutes according to 0.01%, 0.02%, 0.03%, 0.04% ... progressive concentration only is 0.05ml/ at every turn.Right back foot appears later licking until Cavia porcellus, later to lick the right back histamine phosphate's total amount that is given when sufficient be itch-threshold to occur Cavia porcellus at last, calculates and respectively organize the mean and the standard deviation of Cavia porcellus itch-threshold, and do the t check, relatively each group is asked the significance of difference, inspection level α=0.05.
(3) result: as seen by table 11, wood invention medicament capsule tries 1.6,3.2g crude drug in whole/kg dosage group and model group comparison, its itch-threshold significantly improves (P＜0.05 or 0.01), and 0.8g that medicament capsule of the present invention tries (crude drug in whole)/kg group also has certain effect trend.Show that medicament capsule of the present invention has obvious inhibitory action to the local pruritus reaction that Cavia porcellus histamine causes.
Table 11 medicament capsule of the present invention causes the influence of itching to Cavia porcellus histamine part
Compare (t check or t ' check) with model group: * P＞0.05; * P＜0.05; * * P＜0.01.
2, medicine of the present invention causes the influence of mice whole body pruritus to dextran
(1) purpose: dextran iv mice can cause that the whole body pruritus causes mice to produce the pruritus action, and this model can be observed the effect of this product antagonism mice whole body pruritus.
(2) method: 60 of Kunming mouses, male and female half and half, body weight 18～22g is divided into 6 groups at random by body weight and sex, 10 every group.If negative control (solvent control) and positive controls (chlorpheniramine maleate tablets) are subjected to 3 dosage groups of reagent thing (1,2,4g crude drug in whole/kg, be equivalent to 5,10,20 times of clinical every day of recommended dose respectively).Adopt not isoconcentration medicinal liquid gastric infusion (matched group is given distilled water) of isometric(al), be subjected to that 3 concentration of reagent thing are respectively 10,20,40g crude drug in whole/dl, chlorphenamine concentration is 0.02g/dl, and irritating the stomach volume is the 0.1ml/10g body weight, every day 1 time, for three days on end.1h after the last administration, each organizes equal tail vein injection 0.025% dextran physiological salt liquid 0.05ml/10gBW (1.25mg/kg, except the matched group), scratch head, rear solid end of fore paw occur with mice and scratch that to sting each position of whole body be the pruritus indication for trunk, mouth, write down mice pruritus number of times and mean and the standard deviation of pruritus persistent period and average each pruritus persistent period in 20 minutes.The swelling degree significance of difference between the result relatively respectively organizes with the t check, inspection level α=0.05.
(3) result: as seen by table 12, medicament capsule of the present invention tries 2,4g crude drug in whole/kg dosage group and model group relatively, its pruritus number of times and pruritus persistent period are all significantly reduced (P＜0.05 or 0.01), 1g that medicament capsule of the present invention tries (crude drug in whole)/kg group also has certain effect trend, is certain dose-effect relationship.Show that medicament capsule of the present invention causes mice whole body pruritus to dextran antagonism is arranged.
Table 12 medicament capsule of the present invention causes the influence of mice whole body pruritus to dextran
| Group | Number of animals (only) | Dosage (g crude drug in whole/kg * d) | Pruritus number of times (inferior) | The pruritus persistent period (second) | Pruritus persistent period/pruritus number of times (second/time) |
| Matched group model group medicine of the present invention medicine of the present invention medicine chlorphenamine of the present invention | 10 10 10 10 10 10 | - - 1×3 2×3 4×3 1mg/kg×3 | 9.40±3.66 *** 30.80±12.45 25.10±12.33 * 17.10±8.43 *** 19.00±9.78 ** 17.00±7.85 *** | 70.00±35.65 *** 243.10±103.39 197.20±96.51 * 162.40±56.12 ** 151.50±74.09 ** 140.30±74.76 ** | 7.93±3.73 * 8.32±3.45 8.49±3.63 * 11.53±7.30 * 8.68±4.81 * 8.78±4.96 * |
Compare (t check) with model group: * P＞0.05; * P＜0.05; * * P＜0.01.
The 7 medicine antibacterial effects of the present invention of test example
(1) purpose: treatment by Chinese herbs eczema, its antibacterial action are important steps.So select for use the bacterial strain relevant to carry out in-vitro antibacterial test etc. with eczema.
(2) method:
Prepared by the reagent thing: accurately take by weighing capsule content medicated powder (1g medicated powder=3.34g crude drug in whole) 2.00g with weighting procedure, add sterile distilled water 29.3ml and make into suspension, concentration is 64mg/ml, and boiling water bath boils and killed assorted bacterium in 10 minutes.Get 0.5ml during experiment with aseptic MH meat soup (group B streptococcus with serum broth, Candida albicans Sha Shi meat soup) the doubly dilution of opposing, make that concentration series is 32,16,8,4,2,1,0.5,0.25mg/ml.
Test bacterial origin: select the reference culture and clinical isolates strain 16 strains of dermatitis and eczema common pathogen.Comprise staphylococcus aureus ATCC 6538, the staphylococcus aureus clinical strain, staphylococcus epidermidis ATCC 12228, the staphylococcus epidermidis clinical strain, escherichia coli ATCC 25922, the escherichia coli clinical strain, bacillus pyocyaneus ATCC27853, the bacillus pyocyaneus clinical strain, group B streptococcus ATCC19615, the group B streptococcus clinical strain, Klebsiella pneumonia ATCC 13883, the Klebsiella pneumonia clinical strain, Candida albicans ATCC10231, the Candida albicans clinical strain, bacillus subtilis ATCC9372 (brood body), the shigella flexneri clinical strain.Wherein internationally recognized quality control standard bacterial strain is provided by Sichuan Province epidemic prevention station antibacterial section.All the other clinical strains derive from the isolating recently clinical strains of People's Hospital, Sichuan Prov. or Sichuan Province's epidemic prevention station.
The preparation of test organisms liquid: test strain is inoculated in plain agar flat board, sabouraud's agar flat board or blood plate by nutritional need, cultivated 18 hours for 37 ℃, scrape the lawn that takes a morsel and be emulsifiable in MH meat soup (group B streptococcus serum broth, Candida albicans Sha Shi meat soup), than its concentration of turbid correction, do 100,000 times of dilutions again, make final concentration be equivalent to 104cfu/ml.
MIC measures: adopt test tube liquid doubling dilution.The every pipe dosage of test organisms liquid 0.5ml is put 37 ℃ of cultivations behind the mixing, next day observed result.After adding bacterium, investigational agent is equivalent to 1 times of redilution, and concentration series is 16,8,4,2,1,0.5,0.25,0.125mg/ml.The least concentration of bacteria growing inhibiting is MIC.
MBC measures: adopt dull and stereotyped viable bacteria counting method.Promptly measure MIC earlier, will not see successively that again the culture of respectively managing of bacterial growth is drawn 0.1ml respectively and is inoculated in flat board by nutritional need, cultivated 18～24 hours for 37 ℃ again, the lowest concentration of drug of clump count on the flat board＜5 is minimal bactericidal concentration.
Do contrast of bacterium liquid meat soup and investigational agent blank, the contrast of positive drug JIEERYIN XIYE simultaneously.
External minimal inhibitory concentration of table 13 medicament capsule of the present invention (MIC) and minimal bactericidal concentration (MBC) measurement result
| Experimental strain | Medicament capsule of the present invention (the mg content/ml) | JIEERYIN XIYE (minimum extension rate) | ||
| MIC | MBC | MIC | MBC | |
| Staphylococcus aureus ATCC 6538 | 0.5 | 4 | 1∶160 | 1∶80 |
| The staphylococcus aureus clinical strain | 1 | 4 | 1∶40 | 1∶20 |
| Staphylococcus epidermidis ATCC 12228 | 0.5 | 2 | 1∶160 | 1∶40 |
| The staphylococcus epidermidis clinical strain | 1 | 2 | 1∶160 | 1∶40 |
| Escherichia coli ATCC 25922 | 16 | ＞16 | 1∶80 | 1∶5 |
| The escherichia coli clinical strain | 16 | ＞16 | 1∶80 | 1∶5 |
| Bacillus pyocyaneus ATCC27853 | 8 | ＞16 | 1∶40 | 1∶1 |
| The bacillus pyocyaneus clinical strain | ＞16 | ＞16 | 1∶40 | 1∶1 |
| Group B streptococcus ATCC19615 | 2 | 4 | 1∶160 | 1∶80 |
| The group B streptococcus clinical strain | 4 | 8 | 1∶160 | 1∶80 |
| Klebsiella pneumonia ATCC13883 | 0.5 | 1 | 1∶160 | 1∶80 |
| The Klebsiella pneumonia clinical strain | 16 | ＞16 | 1∶160 | 1∶80 |
| Candida albicans ATCC10231 | 4 | 16 | 1∶80 | 1∶20 |
| The Candida albicans clinical strain | ＞16 | ＞16 | 1∶80 | 1∶5 |
| Bacillus subtilis ATCC9372 (brood body) | 0.5 | 2 | 1∶80 | 1∶20 |
| The shigella flexneri clinical strain | ＞16 | ＞16 | 1∶40 | 1∶10 |
(3) result: by table 13 as seen, medicament capsule of the present invention all has certain inhibition or killing action to dermatitis and eczema common pathogen staphylococcus aureus, staphylococcus epidermidis, escherichia coli, bacillus pyocyaneus, group B streptococcus, Klebsiella pneumonia, Candida albicans, bacillus subtilis (brood body), shigella flexneri.
Conclusion (of pressure testing):
1, medicament capsule gastric infusion xylol of the present invention causes the chmice acute ear swelling, the carrageenin local injection brings out all significantly antagonisms of rat foot claw swelling, can the mice dimethylbenzene capillary of skin permeability that anti-inflammatory causes be increased, show that this product has sure antiinflammatory action to the inflammation initial stage.2, medicament capsule of the present invention has obvious inhibitory action to rat passive cutaneous anaphylaxis, PCA of the same race, there are better inhibitory action and mice intradermal injection delayed hypersensitivity that remarkable inhibitory action is all arranged to DNCB induced mice contact skin delayed skin hypersensitivity, have anti-allergic effects.3, medicament capsule of the present invention local pruritus reaction that Cavia porcellus histamine is caused and dextran cause mice whole body pruritus and all have obvious inhibitory action, have itching-relieving action.4, medicament capsule of the present invention has certain inhibition or killing action to dermatitis and eczema common pathogen staphylococcus aureus, staphylococcus epidermidis, escherichia coli, bacillus pyocyaneus, group B streptococcus, Klebsiella pneumonia, Candida albicans, bacillus subtilis (brood body), shigella flexneri, has bacteriostasis.Above result shows, medicament capsule of the present invention has anti-inflammation detumescence, antiallergic, many-sided pharmacological action relevant with acute eczema and dermatitis in treatment such as antipruritic, antibacterial.
To sum up, medicine of the present invention has significant drug effect at treatment eczema.
Claims (8)
1. pharmaceutical composition for the treatment of eczema, it is characterized in that: it is the preparation that is prepared from by the following weight proportion raw material:
1 part of Radix Scutellariae, Radix Gentianae 1-2 part.
2. the pharmaceutical composition of treatment eczema according to claim 1 is characterized in that: it is the medicament that is prepared from by the following weight proportion raw material:
1 part of Radix Scutellariae, 1 part of Radix Gentianae.
3. the pharmaceutical composition of treatment eczema according to claim 1 and 2 is characterized in that: it is to be active component by the water of Radix Scutellariae, Radix Gentianae or 40% above ethanol extraction, adds the medicament that acceptable accessories is prepared from.
4. the pharmaceutical composition of treatment eczema according to claim 3 is characterized in that: described medicament is: capsule, tablet, pill, oral liquid or granule.
5. method for preparing the pharmaceutical composition of each described treatment eczema of claim 1-4, it comprises the steps:
A, get it filled material Radix Scutellariae, Radix Gentianae;
B, adding decocting boil, and perhaps adopt the ethanol extraction more than 40%, reclaim ethanol, filter, and drying adds acceptable accessories and is prepared into medicament pharmaceutically commonly used.
6. the method for the pharmaceutical composition of treatment eczema according to claim 5 is characterized in that: the described concentration of alcohol of b step is: 60% ethanol.
7. the purposes of the described pharmaceutical composition of claim 1 in the medicine of preparation treatment eczema.
8. the purposes of the described pharmaceutical composition of claim 1 in the medicine of preparation treatment acute dermatitis.
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| CN113398164A (en) * | 2021-06-10 | 2021-09-17 | 桂林医学院 | Compound gentian gel for treating infantile eczema and dermatitis |
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