Effects Of Clopidogrel And Atorvastatin On Gene Expression Profiles And Bioinformatic Analysis And Effect Of Atorvastatin On The Pharmacokinetics Of Clopidogrel

Objective: To identify the drug-drug interaction of clopidogrel and atorvastatin based onbioinformatic analysis, pharmacokinetics and retrospective cohort study.Methods:1. Human umbilical vein endothelial cell line EA.hy926were treated with10μM ofatorvastatin or10μM of clopidogrel carboxylic acid for24h, then the cell-based microarraywere performed. GSEA, DAVID and the cMap database were used to analysis the geneexpression data. The target differenced genes of atorvastatin and clopidogrel carboxylicacid were validated by real-time polymerase and Weston-blot.2. In enrolled24healthy volunteers,12patients in each group were treated withclopidogrel75mg for1week or atorvastatin40mg plus clopidogrel75mg for1week, aftera4-week washout period, they received the alternative drug. Blood samples forpharmacokinetic assessments were collected. Concentrations of the active metabolite ofclopidogrel were measured using a validated LC-MS/MS method.3. The study cohort was defined as all patients≥60years old hospitalized for CAD whowere prescribed clopidogrel between January2001and february2011. A total of1021patients were enrolled,187patients were prescribed clopidogrel and843patients wereadministrated clopidogrel combined with statins. The primary endpoint was all causedeath,the secondly endpoint were nonfatal myocardial infarction (MI), and hospitalizationfor unstable angina, stroke, transient ischemic attack, or repeat revascularization (PCI orcoronary artery bypass graft).Results:1. The bioinformatics analysis using GSEA and DAVID revealed that Kruppel-like factors(KLFs) and cell-cycle related genes are the genes most significantly affected byatorvastatin treatment. the cMap analysis suggested that atorvastatin acted as an HDACinhibitor and similar to resveratrol. The genes belonged to Toll-like receptor, Nod-like receptor and IL-6signaling pathway were up-regulated by clopidogrel carboxylic acid,which was similar to15d-PGJ2by cMap analysis. The target genes were validated byreal-time PCR and Weston-blot. The results were consistent with the microarray data.2. Twenty three of the24enrolled subjects completed the study. The mean AUC0_∝ofclopidogrel active metabolite in clopidogrel group was83.12%(19.63±6.32vs23.62±7.65ng/ml·h-1, p<0.01) of clopidogrel plus atorvastatin group. There was no ststisticallydifference in t1/2(0.54±0.19VS0.53±0.21.P=0.78). the Cmax(20.04±8.20VS16.84±5.85,P=0.005) and oral clearance(3.67±1.49VS4.34±1.99,P=0.002)were significantlyaffected by coadministration of atorvastatin.3. Among clopidogrel group and clopidogrel with statins group, the incidence density ofdeath was6.86per1thousand and3.18per1thousand respectively, The crude RR2.15(95%CI:1.39-3.33), and there was statistical significance (χ2=3.53, P<0.01). The twogroups were1:1matched after propensity score matching, clopidogrel coadministratedwith statins showed significant decrease in all cause death, RR=0.42(95%CI0.19-0.93),χ2=7.23, P<0.01. No significant difference was oberserved in death or compositethromboembolic events between statins via different cytochrome P450pathway.Conclusion:1. Atorvastatin played the anti-atherosclerotic role through multiple mechanisms. Themechanism may be similar to the HDAC inhibitors and resveratrol; clopidogrel carboxylicacid increases inflammatory gene pathway, and was similar to the mechanism of15d-PGJ2.From gene expression profiles analysis, there was no significant antagonism betweenatorvastatin and clopidogrel carboxylic acid.2. The pharmacokinetics parameters of clopidogrel active metabolite were significantlydecreased by coadministration of atorvastatin.3. clopidogrel with statin can decrease the mortality of elderly CAD patients comparedwith clopidogrel without statin, There wasn't statistical significance betweenCYP3A4-metabolized statins or non CYP3A4-metabolized statins in mortality andcomposite endpoint events.