Study On PK And PD Of Compatibility Of YTG Prescription Effective Parts

Objective This thesis had chosen the representative main components of YTG prescription effective parts,simultaneously,establised HPLC-DAD method to determine the mass concentrations of puerarin,ferulic acid and TMP in rat plasma.To research the pharmacokinetic characteristics and antioxidant efficacy of main representative components in vivo after MCAO rats taking main effective parts orthogonal compatibility orally.Based on these,we used the intelligent mathematical model to establish the PK-PD model of YTG prescription activity parts,which can provide references for the studies on pharmacokinetic and pharmacodynamic of traditional Chinese medicine compound prescriptions.Methods(1)We established HPLC-DAD methods for determination:the MCAO model was built in rats,and all of the rats were given the effective parts by oral administration(total alkaloid,total flavonoids,total saponins and total phenolic acids)of YTG prescription,and then we obtained blood samples from eye sockets.After pretreated,the blood samples were measured by the reversed-phase column of Agilent Eclipse XDB-C18(4.6 mm × 150 mm,5μm)with 1 mL·min-1 flow rate,column temperature 30℃ and eluted with methanol-0.1%formic acid as mobile phase in gradient mode.Detection wavelengths were 280 nm for puerarin,ferulic acid and TMP.Internal standard method was used as quantitative determination.(2)Pharmacokinetic studies:By using the orthogonal design L9(34)to research the main effective parts(total alkaloid,total flavonoids,total saponins and total phenolic acids)of YTG prescription,9 different dosage ratios combinations were formed,which were used for oral administration in MCAO rats to determine the concentration of puerarin,ferulic acid and TMP in rat plasma at different time points.DAS 3.2.6 software was used to fit the pharmacokinetic parameters of non-compartmental model,and use the total quantum statistical moment analysis method and comprehensive evaluation method to evaluate the total pharmacokinetic characteristics.(3)Pharmacodynamic and PK-PD studies:On the basis of orthogonal combination of nine groups,the normal group and model group were set up,and blood samples were took from eye socket at time points of pharmacokinetic research.The contents of SOD and CAT were determined by ELISA kits.Finally,the two parts of data were simultaneously studied in pharmacokinetic and pharmacodynamics.Using DAS 3.2.6 software to fit the PK-PD model and get the quantitative equations between drug concentration and efficacy.And using Mat lab to build concentration-time-effect of 3D model diagram.Results(1)HPLC-DAD determination method results showed that:The method was successfully established to quantitatively analyze the representative main components(puerarin,ferulic acid and TMP)of YTG prescription effective parts.The linear ranges of puerarin,ferulic acid and TMP were 0.4～80 mg·L-1(R2=0.999 8),1.2～100 mg·L-1(R2=0.9999),0.04～20 mg·L-1(R2=0.998 0)respectively.The mean recoveries were between 85%and 115%and the RSD of intra-day and inter-day were less than 10%.The results of stability met the requirements for biopharmaceutical analysis.(2)The pharmacokinetic results revealed that:There were different pharmacokinetic characteristics of the representative main components(puerarin,ferulic acid and TMP)of YTG prescription effective parts after orthogonal compatibility in rats.Different compatibility had a great influence on each parameter by the total quantum statistical moment analysis and comprehensive evaluation methods.Flavones had the strongest effect on the total AUC with a significantly difference(P<0.05).A3B3C3D3 was the preferred dose combination for AUC.Phenolic acids had the strongest effect on the MRT with a significantly difference(P<0.05).A3B1C2D3 was the preferred dose combination for MRT.Phenolic acids had the strongest effect on the comprehensive evaluation method.A3B3C2D3 was the preferred dose combination for the comprehensive evaluation method.(3)Pharmacodynamic and PK-PD results demonstrated that:The orthogonal compatibility of YTG prescription effective parts could inhibit the reduction of oxidation indexes such as SOD and CAT.Based on the analysis of significant difference and the overall trend,Com7,Com8,Com9 enhanced the vitality of SOD;Com3,Com8,Com9 elevated the vitality of the CAT.The PK and PD models were adopted Sigmoid-Emax model,and the fitting results had a good correlation with the measured data.The R values were more than 0.85.Conclusion(1)The HPLC-DAD method had a high specificity,good degree of separation,simple and quick operating,and the analysis time is appropriate.It suits for the pharmacokinetics studies on the simultaneous determination of puerarin,ferulic acid and TMP in vivo.(2)Compatibility of YTG prescription activity parts had a certain influence on their pharmacokinetic behaviors in MCAO rats.The total quantum statistical moment analysis and comprehensive evaluation method could combine the multi-component pharmacokinetic parameters and express the total pharmacokinetic behaviors.The total quantum statistical moment analysis and comprehensive evaluation method can be used to study the pharmacokinetics of multi-component traditional Chinese medicine(TCM)prescriptions.(3)The effects of YTG prescription against cerebral I/R injury were closely related to the antioxidant properties of the effective parts and constituents.PK-PD model can be used to predict and evaluate the correlation between pharmacokinetics and pharmacodynamics of TCM.Compatibility of YTG prescription activity parts were contributed to optimizing combination drugs screening and evaluating the overall effect of the prescription drugs.It also adequately reflected the thinking mode of traditional Chinese medicine on prescription compatibility.