泰妙菌素对人CYP3A4、CYP1A2、CYP2E1、CYP2D6酶活性的影响

doi:10.16431/j.cnki.1671-7236.2020.04.034

Abstract

Abstract: In this study,the in vitro CYP450 incubation assays were established using human liver microsomes (HLMs) to investigate the effects of Tiamulin on the metabolism of four substrate probes Testosterone,Phenacetin,Chlorzoxazone,and Dextromethorphan hydrobromide which reflect the effect of Tiamulin on CYP3A4,CYP1A2,CYP2E1 and CYP2D6 activity.Experiments were performed in 96-well plates with final volume of 100 μL in each well.The study consisted of three groups:a drug test group,a positive control group (without NADPH) and a negative control group (without CYP450 inhibitor).After incubation were terminated,the concentrations of probe substrates in each well were analyzed by LC-MS/MS internal standard method.The percentage of test drug metabolic was calculated from the ratio of the amount of drug metabolized of the drug test group and that of the negative control group.The percentage of test drug metabolic was plotted versus the logarithm of drugs concentration and the IC₅₀ values of test drugs were obtained from the graph using Graphpad prism 6.0.Multiple incubation time points were set to investigate the effect of the incubation time on the IC₅₀ value of Tiamulin against CYP3A4.The results showed that the IC₅₀ value of Ketoconazole against CYP3A4 was 0.044 μmol/L,the IC₅₀ value of α-Naphthoflavones against CYP1A2 was 0.030 μmol/L,the IC₅₀ value of 4-Methylpyrazole against CYP2E1 was 0.022 μmol/L and the IC₅₀ value of Quinidine against CYP2D6 was 0.096 μmol/L.The IC₅₀ values of Tiamulin against CYP1A2 and CYP2D6 were higher than 50 μmol/L.The IC₅₀ values of Tiamulin against CYP3A4 and CYP2E1 were 1.609 and 0.045 μmol/L,respectively.As the incubation time of Tiamulin and CYP3A4 prolonged from 10 min to 50 min,the IC₅₀ value of Tiamulin against CYP3A4 was increased from 1.609 μmol/L to 8.657 μmol/L.For the IC₅₀ values of the recognized inhibitors were consistent with the reference values,it could be concluded that the in vitro CYP450 incubation assays were constructed successfully.Tiamulin possessed no inhibition against CYP1A2 and CYP2D6,while existed a high inhibitory effect on CYP2E1 and CYP3A4.Tiamulin might be developed as a reversible inhibitor against CYP3A4 in the future.

Key words: Tiamulin; liver microsomes; CYP3A4; CYP2E1; CYP1A2; CYP2D6; IC₅₀ value

CLC Number:

S859.7

CHEN Fang, GAO Hong, LI Bo, ZUO Xiangyi, JIN Zhen, TANG Youzhi. Effects of Taimulin on Human CYP3A4,CYP1A2,CYP2E1 and CYP2D6 Activity[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1267-1273.

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Effects of Taimulin on Human CYP3A4,CYP1A2,CYP2E1 and CYP2D6 Activity

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