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目的:研究mPEG-PLGA聚合物对大鼠肝CYP450酶活性的影响。方法:采用芘荧光探针技术测定mPEG-PLGA聚合物的临界胶束浓度(CMC),并用薄膜分散法制备mPEG-PLGA聚合物胶束;分别采用大鼠肝微粒体和原代大鼠肝细胞模型,将mPEG-PLGA聚合物溶液与CYP450酶特异性探针底物孵育,经液-质联用技术(LC-MS/MS)检测代谢产物,通过比较代谢产物生成量的变化,评价聚合物对CYP450酶的抑制和诱导作用。结果:mPEG-PLGA聚合物的CMC为5.37μg·mL-1;mPEG-PLGA聚合物浓度低于CMC时对大鼠肝CYP450酶无抑制作用;当聚合物胶束质量浓度≥100μg·mL-1时显示有抑制作用,且呈现浓度依赖性。mPEG-PLGA聚合物对CYP1A1/B2、CYP1A2、CYP2B1、CYP2C6、CYP2C11、CYP2D2、CYP3A1/2(底物为咪达唑仑)、CYP3A1/2(底物为睾酮)7种酶亚型的IC50值分别为1.13,2.37,2.38,1.77,1.32,1.17,1.12,0.91 mg·mL-1,其中对CYP3A1/2的抑制作用最为明显。mPEG-PLGA聚合物在0.1~1 000μg·mL-1质量浓度范围对CYP1A2酶均有诱导作用,且在高质量浓度1 000μg·mL-1时最强(P<0.001);而该聚合物仅在0.1μg·mL-1时对CYP2B1、CYP2C6和CYP3A1/2酶呈现诱导作用(P<0.05)。结论:mPEG-PLGA聚合物对大鼠肝脏CYP450酶亚型活性有一定的抑制和诱导作用,其抑制和诱导作用程度与酶亚型种类和聚合物浓度有关。
OBJECTIVE: To study the effect of mPEG-PLGA polymer on liver CYP450 enzyme activity in rats. Methods: The critical micelle concentration (CMC) of mPEG-PLGA polymer was determined by pyrene fluorescence probe technique and the mPEG-PLGA polymer micelles were prepared by thin film dispersion method. The rat liver microsomes and primary rat hepatocytes Model, the mPEG-PLGA polymer solution was incubated with CYP450 enzyme-specific probe substrate. The metabolites were detected by liquid chromatography-mass spectrometry (LC-MS / MS), and the changes of metabolites production Inhibition and induction of CYP450 enzyme. Results: The CMC of mPEG-PLGA polymer was 5.37μg · mL-1. When the concentration of mPEG-PLGA polymer was lower than that of CMC, there was no inhibitory effect on rat liver CYP450 enzyme. When the concentration of polymer micelles was ≥100μg · mL-1 Showed inhibition, and showed a concentration-dependent. IC50 of mPEG-PLGA polymer for seven enzyme subtypes of CYP1A1 / B2, CYP1A2, CYP2B1, CYP2C6, CYP2C11, CYP2D2, CYP3A1 / 2 with midazolam substrate and CYP3A1 / 2 with testosterone substrate Respectively, 1.13,2.37,2.38,1.77,1.32,1.17,1.12,0.91 mg · mL-1, of which the most obvious inhibition of CYP3A1 / 2. The mPEG-PLGA polymer induced the CYP1A2 enzyme in the concentration range of 0.1-1000μg · mL-1 and was the strongest at the concentration of 1 000μg · mL-1 (P <0.001) At 0.1μg · mL-1, CYP2B1, CYP2C6 and CYP3A1 / 2 showed induction (P <0.05). CONCLUSION: The mPEG-PLGA polymer can inhibit and induce CYP450 enzyme activity in rat liver. The degree of inhibition and induction of mPEG-PLGA polymer is related to the enzyme subtype and polymer concentration.