| YiQiFuMai injection(YQFM)is a modified preparation derived from a famous Traditional Chinese medicine(TCM)formula Sheng-Mai-San,which is included in a book named "Medicine Origin"(written by Yuansu Zhang in AD 1186).YQFM is extracted from three kinds of crude drugs:Panax ginseng C.A.MEY.,Ophiopogon japonicus(Thunb.)KER-GAWL and Schisandra chinensis(Turcz.)BAILL(1:3:1.5).According to the theory of TCM,this formulation benefits "qi”,nourishes "yin",prevents exhaustion,and replenishes bodily fluids.Tasly Holding Group has successfully invented Sheng-mai injection in 1970s supported by the 7th five years programs for science and technology development of China,and then produced YQFM in 1999 supported by National modernization project of Chinese traditional medicine.It was launched to the market in 2007 and considered to be curative in treating cardiovascular diseases.Clopidogrel(CPG),a thienopyridine antiplatelet agent,was widely used in preventing and treating heart,brain and other artery circulation obstacles diseases caused by high platelet aggregation state.As combination use of herbs and drugs is a very common approach in clinical therapy to improve drug efficacy or avoid side effects,will it be more efficiency or safer in co-adiministrated YQFM with CPG?In our study,the effect of YQFM on pharmacokinetics and pharmacodynamics of CPG was investigated in rats to provide guidance for clinical use.Firstly,we developed an LC-MS/MS method for the determination of clopidogrel carboxylic acid(SR26334).Mass spectrometric detection was carried out on a Surveyor HPLC System coupled to a Thermo Finnigan TSQ Quantum triple quadrupole mass spectrometer(Thermo Finnigan,USA).The analytical column was Agilent Eclipse XDB-C8(4.6mm×150mm,3.5μm).The mobile phase was a mixture of 0.1%formic acid:methol(5:95,v/v)and pumped at a flow rate of 0.5 mL/min,with an analysis time was 5 min.Linearity,precision,accuracy,recovery and stability were examined for quantitative analysis in vivo pharmacokinetics.This method is simple,fast,accurate and available for the determination of clopidogrel carboxylic acid.The standard curve is Y=0.970085X+0.014395,linearity over a concentration range from 0.02 to 20 μg/mL was corroborated greater than 0.991,with the lower limit of quantification(LLOQ)of 5 ng/mL.Specificity,precision,accuracy,extraction rate and stability all meet the requirements of the analysis.Secondly,we investigate the effect YQFM on pharmacokinetics of CPG in rats,PK study was carried out by the established method.Two groups of Wistar rats(n=6/group)received CPG alone and CPG in combination with YQFM,respectively.The CPG group animals were orally administrated with CPG at dose of 15 mg/kg(human equivalent dosage)and combination group animals were received YQFM at dose of 542 mg/kg(human equivalent dosage)by intravenous injection 1 h prior to CPG(15 mg/kg)administration.In single dose experiment,whole blood samples were collected through cannula at a series of time points within 48 h after dosing CPG.In multiple doses experiment,the rats were treated for consecutive 7 days.Following dosing on Day 7,whole blood samples were collected through cannula at a series of time points within 48 h after dosing CPG.Plasma samples were prepared and a 10μL aliquot was injected into the chromatographic column.Parameters like AUC0-t、Cmax、Tmax、T1/2 of CPG were not altered by concomitant application of YQFM and CPG(P>0.05).YQFM had no significant effect on pharmacokinetics of CPG.Thirdly,we reflect the efficiency of extrinsic clotting pathway,intrinsic coagulation phase in plasma and evaluate the risk of bleeding.Bleeding time(BT)and clotting time(PT and APTT)were measured in pharmacodynamic study.In clotting time experiment,the rat dosing were same as PK study and plasma samples were collected for measuring PT and APTT.All procedures were obeyed to introduction of reagents.In bleeding time experiment,the rat dosing were same as PK study and a control group(n=6)treated without drug was added as a reference.Tails of the animals were transected 2 mm from the lip by a disposable surgical blade and immersed into 10 mL normal saline at 37℃ by a water bath immediately.BT of each rat from incision to termination was recorded.No obvious differences were found in measuring BT,PT and APTT between CPG group and combination group.YQFM does not affect the coagulation properties and adverse effects of CPG,Pre-treatment of YQFM with clopidogrel does not affect the pharmacokinetics and pharmacodynamics of clopidogrel,that is to say,the combination of YQFM with CPG is safe in rats.Whether there is interaction for human in vivo still needs further research. |
