Study On Compatibility Screening, Pharmacodynamic Action And Pharmacokinetics Of Traditional Chinese Medicine Compound That Was Studied Out By Ourselves

The study object of our topic is traditional Chinese medicine compound that was studied out by ourselves (effective parts of traditional Chinese medicine), according to respective toxicity and drug efficacy study, screen perfect matching by uniformity design method and study it's pharmacodynamics and initial pharmacokinetics to illuminate and approach the modern traditional Chinese medicine rationality of compatibility and mechanism of action. ( the traditional Chinese medicine compound is made of effective parts of magnolia bark and fructus cnidii, it's function are drying dampness to eliminate phlegm, warming kidney and absorbing Qi, descending qi and relieving asthma. It is used to treat cough with asthma, chest distress and gastric cavity turgor, much expirium and few airintake, breathe heavly when moving, lassitude and debilitation, soreness of waist and weak of knees that was caused by phlegmatic hygrosis accumulation inside, chronic disease involving kidney and kidney failing to promote inspiration).Part One. study on screening of effective parts compatibility of magnoliae cortex and fructusTo determine LD50 of magnoliae cortex extractive, fructus extractive and their clathrate, determine their dosage limit by pharmacodynamics experiment. According to uniformity design principle, to carry out study on screening prescription compatibility.1. study on acute toxicity of single medical material extractive and it's β cyclodextrin inclusion compoundMake linear regression equation with logarithmic dose and death rate's probability unit.Fructus: y=8.3171x+2.5146, r=0.9937,magnoliae cortex: y=8.4248x+0.5641, r=0.9977,,It shows that fructus clathrate's toxicity LD50=2.279g/kg has not noticeable difference compared with fructus clathrate that's LD50 = 2.05g/kg, magnoliae cortex's toxicity is weaker than magnoliae cortex fructus extractive that's LD50=3.35g/kg.2. pharmacodynamics study of magnoliae cortex and fructus' effective parts1) the results of antagonistic effect on cavia cobaya asthma caused by histamine phosphate show that both magnoliae cortex extractive and fructus extractive can lengthen delitescence of beta-imidazolyle thylamine's leading to wheeze, fructus' effective dose is 0.2g/kg and magnoliae cortex's is 0.3g/kg by screening, but magnoliae cortex's delitescence is a little lower than aminofilina.2) We observed the antagonistic effect on asthma caused by 0.1 % histamine phosphate and 2% acecolex mixed liquor. The results showed that both both magnoliae cortex extractive and fructus extractive can rivalry bronchus convulsion in different degree, obviously lengthen delitescence, and have significant difference (p<0.05) or highly significant difference (p<0.01) ,but the dosage are fairly great.3) The experimental result that antagonistic action on allergic asthma caused by egg albumin spraying shows that the two medical materials' extractive both can lengthen delitescence and degrade order of reaction.3. study on screening prescription compatibility by uniformity design method study on screening prescription compatibility by uniformity design method, the resultshows that all compatibilities of different dose extractive can lengthen delitescence leadingto asthma, every formula has significant difference compared with NS control group(p<0.05) .according to standard of most eclipse period extension, least dose ofprescription and better drug action, we determined that prescription 4 is more Idealcompatibility that is magnoliae cortex is 0.20 and fructus is 0.15.Analysis of experiment result of screening prescription by uniformity design method: Make polynary linear regression equation with logarithmic dose and death rate's probability unit(by looking up probit table):Y=1.595×fructus+2.825×magnoliae cortex+4.603×fructus×magnoliae cortex。 We can conclude: 1. from coefficient of fructus×magnoliae cortex we can discern that fructus has most influence on death rate, next is magnoliae cortex. 2 . subst. log(D magnoliae cortex ×5)=log1=0 to regression equation, we made y=5( probit of 50% death rate), get LD50fructus=3.28 g/kg, also we can get LD5magnoliae cortex=13.65 g/kg, both exceed single medicine's LD50. So the compatibility possibly have attenuation in abstracto.Two. Pharmacodynamics study after screening prescription.Adopt isolated organ and whole animal experiment. Result of isolated organ experiment shows, traditional Chinese medicine complex prescription has obvious depressant effect on isolated air tube construction caused by beta-imidazolyle thylamine and acetylcholine to cavia cobaya., IC50 respectively is 17.02ug/ml and20.94 ug/ ml by Bliss procedure.Result of whole animal experiment shows: after screening complex prescription can significantly prolong the delitescence of leading to asthma in cavia cobaya caused by beta-imidazolyle thylamine-acetylcholine by intragastric administration consecutively 7 days with dose of 60 and 120mg/kg, it can reduce animal amount of convulsion. It can significantly prolong convulsion delitescence caused by egg albumen in Cavia procellus and decrease cavia cobaya amount of shock.. After screening the traditional Chinese medicine complex prescription can obviously decrease lung outflow volume of allergized anesthetized rats by intragastric administration consecutively 7 days with dose of 70 and 140mg/kg. The result of bulk testing in mice shows, the traditional Chinese medicine complex prescription has significant depressant effect on licking body reaction in mice caused by 4- amion pyridine, it can significantly increase phenol red eccrisis of air tube in mice. It has significant prolongation in tussis delitescence caused by ammonia water with dose of 50,100 and 200mg/kg by intragastric administration consecutively 7 days. It can significantly delay body weight of renal deficiency in mice caused by cetacort. And itcan significantly increase thoracic gland index, spleen index and cell phagocytosis in immune defect mice caused by hydro-adreson.. It can significantly increase total SOD activity, Mn-SOD activity and Cu · Zn-SOD activity in serum of yang asthenia mice caused by hydro-adreson with intragastnc administration of successive 14 days. It can significantly increase samenblase corpus glandulare prostatae coefficient of demasculinized mice by intragastnc administration of successive 14 days with dose of 200mg/kg and the dose of 50 and 100mg/kg have the tendency of increasing them.Three. pharmacokinetics study of traditional Chinese medicine compoundchromatographic condition: chromato bar: LiChro CART 250-4 C18, 200 × 4.6mm, grain size 5um; mobile phase: methanol: 0.02mol/l of ammonium dihydrogen phosphate aq sol(84: 16) , flow rate is 0.8ml/min; detection wave length: 290nm: column temperature is 25℃; sample size: 20ul. Respectively take quantity sufficient of magnolol, ammidin and osthole control article dissolved by methanol and blank serum sample solution as control article solution, take control article solution, control article added serum and sample solution to detect according to above-mentioned chromatographic condition, appearance time of magnolol is about 6.2min, number of theoretical plates n=7203 and resolving power of prepeak and postpeak is R>1.5. appearance time of ammidin is about 4.3min, number of theoretical plates n=5749 and resolving power of prepeak and postpeak is R>1.5. Appearance time of osthole is about 5.3 min, number of theoretical plates n=7534 and resolving power of prepeak and postpeak is R>1.5.Concoction Process of serum working sample: take 250 μl of serum, add 750 μl of deproteinizing agent(saturated white vitriol WS: acetonitrile=1:9), intensively shake it 1min in whirlpool admixer to mix it, then centrifuge for 10min with 3000rpm,take 40 μl of clear supernatant liquid to filter with filter membrane, at last detect it with HPLC in 290nm.Computation of pharmacokinetics shows, Medication of magnoliae cortex with fructus can significantly heighten maximum concentration, peak time obviously decreased(magnolol) or increased(osthole and ammidin); area under curve obviously increased; absorption demiperiod obviously decreased(magnolol) or increased(osthole and ammidin); elimination half-time obviously prolonged (magnolol) or shortened(osthole andammidin), mean residence time obviously lengthened(magnolol) or shortened(osthole and ammidin).Four .result and discussion1. we adopt uniformity design to screen prescription compatibility of effective parts, the result shows, the fairly Ideal compatibility is fructus is 0.15 and magnolia bark is 0.20, LD50 of compatibility is more than single extractive's, this illustrate that compatibility possibly has attenuation, and drug action exceed virgin single fructus and magnolia bark extractive.Pharmacodynamics study after screening compatibility shows: traditional Chinese medicine complex prescription has significant effect on preventing cough and depressing asthma after screening, it is consistent with traditional Chinese medicine's Medication effect.3. From pharmacokinetics parameter we can know, Cmax of magnolol increased and Tmax shortened, this shows that after Medication of combination concentration of magnolol in blood quickly reach peak, and the fact of T1/2(Ka) obviously shortened also support the result, significant prolongation of T1/2(Ke) shows that after Medication of combination blood drug level not only significantly increased, but also it's annihilation step down, effective blood drug level time prolonged. Opposite to above-mentioned result, after Medication of combination of magnolia bark clathrate and fructus clathrate, blood level of osthole and ammidin also obviously increased, but their Cmax increased very slowly. significant prolongation of T1/2(Ka) also proved that hysteresis of Cmax is clear, and T1/2(Ke) obviously shortened. This means that it can make these two venenosus compound metabolic conversion and rule out from body promptly. Therefore, combination of these two clathrate can get the purpose of synergia and Attenuation, adopt good points and avoid short-comings.Study Result shows, complex prescription compatibility has obvious effects on pharmacal pharmacokinetics parameter, osthole, ammidin and magnolol one another have interaction on pharmacokinetics, this proves compatibility that made up according to Chinese medical science and scientific research experiment is reasonable. From analysis of...