Compound Danshen Dripping Pills Affect The Pharmacokinetics Of Azisartan By Regulating The Expression Of Cytochrome P450 In Rats

Part one Effect of Compound Danshen Dripping Pills on thepharmacokinetics of azilsartan in vivoObjective:To establish a quantitative analysis method for the determination of azisartan in rat plasma,and to evaluate the effects of different doses of compound danshen dripping pills on the pharmacokinetics of azisartan.Methods:Twenty four male Sprague Dawley?SD?rats were randomly divided into four groups:low dose Compound Danshen Dripping Pills?81mg/kg?+azilsartan group;medium dose Compound Danshen Dripping Pills?405 mg/kg?+azilsartan group;high dose Compound Danshen Dripping Pills?810 mg/kg?+azilsartan group;and control group?AZ was applied individually?.All Sprague-Dawley rats were administered with a human equivalent oral dose of AZ?2 mg/kg?.According to the time point of the design,the blood was taken through the orbital vein,and the plasma sample was obtained.The sample was pretreated by acetonitrile protein precipitation method.Chromatographic conditions:Waters XBridge BEH C18 column?2.1x100 mm,2.5?m?;mobile phase:acetonitrile?A?-0.1%formic acid water?B?,gradient elution;flow rate:0.4 m L/min;pre-column equilibrium time:2min;injection volume:3?L.Mass spectrometry conditions:electrospray ionization source?ESI?,positive ion mode detection;multiple reaction monitoring?MRM?mode scan.Non-compartmental model analysis,the main pharmacokinetic parameters of azisartan were calculated by DAS 2.1.1,and SPSS 21.0 software was used to compare the differences of pharmacokinetic parameters of the Compound Danshen Dripping Pills combined with the azisartan group and the control group.Results:Calibration curves for azisartan in rat plasma showed good linearity over the test concentration range of 2–2,000 ng/m L,and the methodologies have been verified to meet the requirements.The pharmacokinetic analysis indicated that the combination groups significantly increased C_max and AUC and decreased CL significantly compared with the control group of azisartan alone.The C_max of azisartan in the low,middle and high dose groups was 1.96,1.97,2.13 times higher than that in the control group,AUC_0-t was 1.78,1.89,2.03 times higher than that in the control group,and AUC_0-?was 1.78,1.95 and 2.03 times higher than that in the control group.CL in the high-dose group was significantly reduced by 4.5 times compared to the control group.Conclusion:In this study,a rapid,sensitive,and highly selective UPLC-MS/MS method was established for determining the azisartan plasma concentration in rats and applied to evaluate the effects of different doses of compound danshen dripping pills on the pharmacokinetic parameters of azisartan.Pharmacokinetic analysis indicated that compound danshen dripping pills could reduce the metabolism of azisartan and significantly increase its concentration in vivo.The experimental results provide a scientific illustration and theoretical basis for the rational use of the two drugs in clinical practice.Part two Effects of compound danshen dripping pills on CYP2B1,Objective:To investigate the changes of m RNA and protein expression levels of compound danshen dripping pills on CYP2B1,CYP2C6 and CYP2C11 in rat liver.This study expounds the effect of compound danshen dripping pills on the metabolism of azilsartan from the perspective of metabolism,and provides a reference for clinical application to ensure that the two can be used safely and reasonably.Methods:The m RNA and protein levels of CYP2B1,CYP2C6,and CYP2C11 in the rat liver were detected by RT-PCR and Western Blot,respectively.Results:Compared with the control group alone,the m RNA and protein expressions of CYP2B1,CYP2C6 and CYP2C11 in the combination group were significantly reduced.Compound danshen dripping pills can inhibit the expression of CYP2B1,CYP2C6 and CYP2C11 m RNA and protein.CYP2B1m RNA was significantly reduced by 1.09-fold,1.37-fold,and 2.00-fold in the low,medium,and high-dose groups;CYP2C6 was 1.65-fold,2.15-fold,and3.36-fold,respectively;CYP2C11 was significantly reduced by 1.20-fold,1.58-fold,and 3.30-fold.The expression level of the protein was consistent with its corresponding m RNA expression,and both were significantly inhibited.Conclusion:We found that compound danshen dripping pills significantly inhibited the m RNA and protein expression of CYP2B1,CYP2C6,and CYP2C11,and the inhibitory effect became more pronounced as the dose increased.Compound Danshen dripping pills may affect the metabolism of azisartan in vivo by inhibiting the activities of CYP2B1,CYP2C6 and CYP2C11.The data indicate potential drug-drug interactions in the combination therapy.Therefore,we should consider personalized dose adjustments in patients.The experimental results also provide a scientific illustration and theoretical basis for the rational use of the two drugs in clinical practice.