The Study Of Drug-drug Interaction Between Ticagrelor And Aspirin Based On UGT Enzyme

Objective:Observed the platelet aggregation rate on the condition of dual antiplatelet therapy(DAPT).The inhibition of uridine diphosphate glucuronide transferase(UGT)enzyme by ticagrelor in vitro was investigated using recombinant UGT isoforms to predict the potential drug-drug interactions between ticagrelor and aspirin.Preliminary explored the increase in the incidence of bleeding caused by ticagrelor.Methods:1.55 patients who underwent percutaneous coronary intervention(PCI)and regularly taken dual antiplatelet drugs from June 2015 to February 2016 in Tianjin Union Medicine Center were divided into ticagrelor group and clopidogrel group according to the difference of P2Y12 receptor inhibitors.The platelet aggregation rate were measured by light transmission aggregometry(LTA)in both groups.2.The UGT isoforms that commonly participated in drug metabolism were selected,namely UGT1A1,UGT1A3,UGT1A7,UGT1A8,UGT1A9,UGT1A10,UGT2B4,UGT2B7,UGT2B15 and UGT2B17,were used to establish the incubation system of glucuronic acid binding reaction in vitro with 4-methyl umbeliferone(4-MU)as non-specific substrate.Ticagrelor was added as an inhibitor in incubation system.The amount of 4-methylumbelliferone-?-D-glucuronide(4-MUG)that was the glucuronic acid binding product of 4-MU was detected by ultra performance liquid chromatography(UPLC)using 7-OH coumarin as internal standard.First,the initial determination of the degree of inhibition was carried out.The enzyme concentration,the substrate concentration and the reaction time were selected according to the previous studies.The amount of 4-MUG produced was measured when there was an inhibitor and no inhibitor,and the residual ratio and the inhibition rate were calculated.Then,the reaction of multiple concentrations of inhibitor were selected to observe the relationship between the inhibitory degree of UGT enzyme and the concentration of ticagrelor,and the half inhibitory concentration(IC50)values were calculated.Four inhibitor concentrations and blank control were selected according to the IC50 values and four 4-MU concentrations were selected for the reaction.Lineweaver–Burk plots were employed to obtain the inhibition type.The second plot of slopes from Lineweaver–Burk plot versus ticagrelor' concentrations was used to calculate the inhibition kinetic parameters(Ki)value.The possibility of inhibition of UGT and drug interactions in vivo was inferred from the ratio of the ticagrelor's concentration in vivo [I] to Ki.Finally,molecular docking was performed using computer software to explain the inhibitory mechanism of ticagrelor to UGT enzyme.Results:1.The platelet aggregation rate induced by arachidonic acid(AA)and adenosine diphosphate(ADP)in the ticagrelor group were lower than those in the clopidogrel group(all P <0.05).2.The inhibitory rates of UGT1A1,UGT1A6,UGT1A7,UGT1A7,UGT1A8,UGT1A9,UGT1A10,UGT2B4,UGT2B7,UGT2B15 and UGT2B17 by 100?M ticagrelor were 74.98%,84.01%,100%,84.66%,100%,98.03%,93.58%,100%,100%,99.99% and 99.32%,respectively.3.The IC50 values of ticagrelor to UGT1A1,UGT1A3,UGT1A6,UGT1A7,UGT1A8,UGT1A9,UGT1A10,UGT2B4,UGT2B7,UGT2B15 and UGT2B17 were:0.043,8.256,12.86,2.472,39.34,0.2994,6.942,36.55,2.118,12.30 and 34.40 ?M,respectively.4.Ticagrelor competitively inhibited UGT1A1,1A3,1A9 and noncompetitively inhibited UGT1A6,1A7,1A10,2B4,2B7,2B15.5.The Ki values of ticagrelor to UGT1A1,UGT 1A3,UGT 1A6,UGT 1A7,UGT 1A8,UGT 1A9,UGT 1A10,UGT 2B4,UGT 2B7 and UGT 2B15 were: 2.05,4.05,1.59,40.32,0.0208,8.63,8.08,2.05 and 4.80 ?M,respectively.6.According to taking 180 mg ticagrelor,the maximum plasma concentration in vivo was 2.34?M.The [I] / Ki of UGT1A1,UGT1A3,UGT1A6,UGT1A7,UGT1A9,UGT1A10,UGT2B4,UGT2B7,UGT2B15 are calculated as follows:1.14,0.58,1.47,0.058,112.5,0.27,0.29,1.14 and 0.48,respectively.[I] / Ki <0.1 for UGT1A7,meant a low degree of likelihood inhibition;0.1 <[I] / Ki <1 for UGT1A3,1A10,2B4,2B15,meant a medium degree of likelihood inhibition;[I] /Ki> 1 for UGT1A1,1A6,1A9,2B7,meant a high degree of likelihood inhibition.7.In silico docking study indicates the binding free energy of 4-MU and ticagrelor towards UGT1A1 were-5.68 and-9.45 kcal/mol,respectively.The binding free energy of 4-MU and ticagrelor towards UGT1A3 were-5.65 and-8.63 kcal/mol,respectively.The binding free energy of 4-MU and ticagrelor towards UGT1A6 were-5.92 and-7.41 kcal/mol,respectively.The binding free energy of 4-MU and ticagrelor towards UGT1A7 were-6.04 and-8.36 kcal/mol,respectively.The binding free energy of 4-MU and ticagrelor towards UGT1A9 were-6.40 and-7.54 kcal/mol,respectively.Conclusions:1.Compared with clopidogrel combined with aspirin,the platelet aggregation rate induced by AA and ADP of ticagrelor combined with aspirin were both lower.2.Ticagrelor could inhibit the activity of UGT1A1,UGT1A3,UGT1A6,UGT1A7,UGT1A8,UGT1A9,UGT1A10,UGT2B4,UGT2B7,UGT2B15 and UGT2B17.Ticagrelor competitively inhibited UGT1A1,1A3,1A9,and noncompetitively inhibited of UGT1A6,1A7,1A10,2B4,2B7,2B15.3.In vivo,ticagrelor is high possible to inhibit UGT1A1,1A6,1A9,2B7,moderately possible to inhibit UGT1A3,1A10,2B4,2B15,and low possible to inhibit UGT1A7.4.Ticagrelor inhibited the activity of UGT isoforms,suggesting that it may inhibit the metabolism of salicylic acid that is aspirin's metabolite.