Study On Evaluating The Bioequivalence Of Two Preparations In Human Body Based On Ezetimibe And Total Ezetimibe Exposure Concentrations

BackgroundCoronary heart disease is a clinical high incidence cardiovascular disease,and its main inducing factor is coronary atherosclerosis.In clinical treatment,statins are mainly used to reduce blood lipid to achieve the therapeutic effect.Ezetimibe is a selective cholesterol absorption inhibitor.Combined with statins,ezetimibe can more effectively regulate the blood lipid level of patients,reduce the dosage of statins and reduce the side effects of statins.There is still a lack of methods to accurately determine the concentrations of ezetimibe and total ezetimibe in human plasma at the same time.Objective（1）To establish a liquid chromatography tandem mass spectrometry（LC-MS/MS）is a method for the determination of ezetimibe and total ezetimibe in human plasma.（2）To investigate the relevant pharmacokinetic parameters and relative bioavailability of ezetimibe test preparation and reference preparation in a single administration,and to evaluate the bioequivalence and safety of the two preparations.MethodsEzetimibe tablets developed and produced by a company were used as the trial preparation R(10 mg / tablet based on C₂₄H₂₁F₂NO₃)and ezetimibe tablets produced by MSD Pharma（Singapore）Pte.Ltd(Ezetrol^（?）)10 mg / tablet was used as reference preparation T for bioequivalence test.Bioequivalence test is a single center,single dose,fasting / postprandial,randomized,open,two preparation,two cycle and double crossover bioequivalence test conducted in healthy adult subjects.48 subjects in fasting group / postprandial group were randomly divided into A（TR）and B（RT）groups according to the screening order.All subjects were given 10 mg of T or R in fasting / postprandial state,and blood samples were collected at the specified time point.Plasma samples were treated by direct precipitation method and post enzymatic precipitation method.A LC-MS / MS method was established for the determination of ezetimibe and total ezetimibe in plasma.Result（1）In the LC-MS / MS method established in this study,the linear range of ezetimibe in plasma is 0.1 ～ 20 ng / ml,and the lower limit of lloq is 0.1 ng / ml;The linear range of total ezetimibe is 1 ～ 200 ng / ml,and the lower limit of lloq is 1 ng / ml.The accuracy of ezetimibe was98.6 ～ 106.6%,the accuracy of total ezetimibe was 93.5 ～ 105.1%,and the RSD of intra batch and inter batch precision was less than 15.00%.The recovery of ezetimibe was 103.8 ～ 108.7%,the recovery of total ezetimibe was 106.7 ～ 109.8%,and the RSD was less than 5.0%.The matrix effect of ezetimibe and total ezetimibe was 70.5 ～ 79.8%,and the RSD was less than 5.00%;The accuracy of 2% hemolysis effect and high-fat matrix effect was 103 ～ 108.6%,and the RSD was less than5.00%.The stability test results showed that the plasma quality control samples of ezetimibe and total ezetimibe were stable under various conditions.The determination of the drug to be tested and the internal standard will not be disturbed by endogenous substances in plasma samples.This method can accurately determine the concentrations of ezetimibe and total ezetimibe in plasma with high specificity and sensitivity.（2）Equivalence testFasting group: a total of 111 subjects were selected in the fasting study,and 48 subjects were enrolled.All 48 subjects completed two cycles of administration and follow-up according to the test scheme.Postprandial group: 115 subjects were screened in the postprandial study,including 49 enrolled,47 completed the test and 2 exfoliated subjects.1)Pharmacokinetic characteristics and bioequivalence evaluationThe pharmacokinetic parameters C_max,AUC_0-t and AUC_0-∞ of the two groups were tested by two-way one-sided t-test after logarithmic conversion.The 90% CI of the geometric mean ratio of C_max,AUC_0-t and AUC_0-∞ was 80.00%～125.00%,which was in line with the bioequivalence evaluation criteria.2)Safety evaluationIn the fasting group,the incidence of adverse events and adverse reactions in the trial preparation group were 29.17%（14 / 48）and 14.58%（7 / 48）;The incidence of adverse events in the reference preparation group was 37.50%（18 / 48）,and the incidence of adverse reactions was16.67%（8 / 48）.In the postprandial group,the incidence of adverse events was29.17%（14 / 48）and the incidence of adverse reactions was 20.83%（10 /48）;The incidence of adverse events in the reference preparation group was 29.79%（14 / 47）,and the incidence of adverse reactions was 17.02%（8 / 47）.The adverse events related to the study drug occurred in this trial were mild and short duration.Most subjects could tolerate them.There were no serious adverse events related to the study drug.The symptoms of most subjects could return to normal by themselves.ConclusionLC-MS / MS has good accuracy,strong specificity and high stability.It can quickly and accurately determine the concentrations of ezetimibe and total ezetimibe in human plasma.According to the analysis and comparison of pharmacokinetic parameters,ezetimibe tablets(10 mg/ tablet based on C₂₄H₂₁F₂NO₃)developed by a company and ezetimibe tablets(Ezetrol^（?）)produced by Hangzhou moshadong Pharmaceutical Co.,Ltd 10 mg / tablet is bioequivalent and safe when administered fasting /postprandial.15 graphs,37 tables and 57 references...