| BackgroundVonoprazan fumarate,a potassium-competitive acid blocker,has the advantage of replacing traditional proton pump inhibitors as a new acid inhibitor.As it has just entered the Chinese market,the study of its drug interactions will be of certain significance for its further clinical application.vonoprazan is mostly metabolized by the cytochrome P450.This study intended to establish a type UPLC-MS/MS method,which could determine six different probe drugs and their metabolites at the same time to present the activity of CYP450 enzyme.Thus,we could estimate the inhibiting effect that vonoprazan impacted on them,providing the clue for its potential drug-drug interactions in clinical settings.MethodsIn the first part of this study,vonoprazan and six probe drugs(phenacetin,bupropion,tolbutamide,dextromethorphan,chlorzoxazone and midazolam),representing six kinds of CYP subtypes(CYP1A2,CYP2B6,CYP2C9,CYP2D6,CYP2E1 and CYP3A4).We established an UPLC-MS/MS method,using it to detect those drug concentrations together at different time points,so that the extent of inhibiting impact could be estimated.Then the concentration of probe drug was also detected in vivo.Rats were randomly assigned into two groups: one was administered six probe drug mixtures and vonoprazan,while the other without vonoprazan.By using UPLC-MS/MS method,we tested the drug concentrations in blood in order to evaluat vonoprazan influence on the activity of target CYP450 subtypes from the in vivo level.ResultsIn vitro microsomal experiments showed that the IC50 values of bupropion,tolbutamide,dextromethorphan and midazolam in rat microsomes were 3.68,16.11,3.43,and 22.12μM,respectively,under the inhibition of vonoprazan,while chlorzoxazone and phenacetin were not affected.In vivo analysis demonstrated that many pharmacokinetic parameters of midazolam,tolbutamide,dextromethorphan and bupropion were significantly different from those of the untreated(P<0.05),while the pharmacokinetic parameters of chlorzoxazone and phenacetin showed no significant differences.It was consistent with the above results in vitro.ConclusionBoth in vitro and in vivo results showed that vonoprazan inhibited the activity of CYP2B6,CYP2C9,CYP2D6,and CYP3A4 those four CYP450 subtype enzymes,suggesting that in clinical applications,vonoprazan may impair the efficiency of drugs that metabolized by cytochrome P450 substrates,which should be pay attention to the possible side effects or changes in curative effectBackgroundIn the first part of the study,vonoprazan was used as an inhibitor to study its effect on the activity of different CYP enzyme subtypes,suggesting that vonoprazan may inhibit drugs metabolized by CYP2B6,CYP2C9,CYP2D6,and CYP3A4 enzymes.However,since vonoprazan itself is mostly metabolized by CYP enzymes,it may also be affected by other drugs that inhibit or induce CYP enzyme activity.Therefore,this part intends to use vonoprazan as a substrate to study the influence of some commonly used clinical drugs on drug metabolism.MethodsThis part is the same as the previous study from both in vivo and in vitro.First,incubate vonoprazan and different drugs together in the in vitro incubation system of rat liver microsomes.UPLC-MS/MS was applied to detect the concentration of vonoprazan metabolite,M1,in order to screen drugs that may produce inhibitory or inducing effects.Drugs with obvious inhibitory effects(inhibition effect greater than 5 times)were further tested for the half inhibitory concentration(IC50).Then three drugs with lower IC50 and higher clinical value were selected for further in vitro inhibition mechanism experiments and in vivo experiments.In vivo experiments group rats into groups of inhibitors for 1 week,and gavage vonoprazan on the last day.UPLC-MS/MS is used to detect the concentrations of vonoprazan prototype and metabolite M1 in blood in order to evaluate the effect of target clinical drug administration on the metabolism of vonoprazan from the rat in vivo level.ResultsIn vitro microsomal experiments were used to preliminarily screen out 10 possible inhibitors among 49 clinical medicines and Chinese herb monomer,and the IC50 was detected to be 21.87μM for nifedipine,16.37μM for amlodipine besylate,and 12μM for simvastatin,21.7μM for lometapide,10.6μM for poziotinib,0.78μM for napabucasin,17.96μM for mocetinostat,24.14μM for rotigotine.There are also three kinds of Chinese medicine monomer: 11.61μM for apigenin,9.46μM for acacetin,1.66μM for imperatorin.Considering the IC50 and the clinical value of combined use with vonoprazan,three drugs,simvastatin,amlodipine,and poziotinib,were selected for in vitro drug mechanism research.The inhibitory mechanisms of the three drugs on vonoprazan were found to be as follows: anti-competitive and non-competitive mixed type,non-competitive and competitive mixed type,and non-competitive and anticompetitive mixed type.Further in vivo experimental analysis showed that simvastatin,amlodipine and poziotinib all have varying degrees of inhibitory effects on the metabolism of vonoprazan carboxylic acid metabolite M1,which is consistent with the in vitro screening results.But for the drug prototype,amlodipine did not have a significant impact on the metabolism of the vonoprazan prototype drug.The Vz/F of the simvastatin group was significantly increased to 3 times compared with the control(P<0.05),and the Cmax was reduced to 53% of the control(P< 0.05),contrary to the hypothetical inhibition of drug prototype metabolism.The AUC of the vonoprazan prototype in the poziotinib group was significantly increased by four times(P<0.01),MRT was prolonged by 2.4 times(P<0.01),and the Cmax of the drug increased by 2.7 times(P<0.01).The corresponding performance of the carboxylic acid metabolite M1 showed that the Cmax was significantly reduced to 29.2% of the control(P<0.01).ConclusionVonoprazan is used as a CYP450 substrate for inhibitor screening studies in vitro and in vivo.Vonoprazan shows good drug metabolism stability.The screened amlodipine may not have obvious clinical inhibition effects.What’s more,simvastatin may even induce prototype metabolism.The anti-tumor drug poziotinib has a significant inhibitory effect on the metabolism of vonoprazan,and the inhibitory mechanism may not be limited to the CYP450 enzyme.All the puzzles need further researchs and verifications. |
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