血脂康抑制HMG-CoA还原酶活性的定量测定研究

发布时间：2018-01-14 14:26

本文关键词：血脂康抑制HMG-CoA还原酶活性的定量测定研究　出处：《中国新药杂志》2015年16期 　论文类型：期刊论文

【摘要】：目的:建立血脂康的活性测定方法,用于血脂康的质量控制研究。方法:建立3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMG-Co A还原酶)酶促反应,即HMG-Co A还原酶量为3.125×10-4U,烟酰胺腺嘌呤二核苷磷酸(NADPH)量为160 nmol·L-1,羟甲基戊二酸单酰辅酶A(HMG-Co A)量为5 nmol·L-1,反应时间为50 min的反应体系,加入洛伐他汀酸系列工作溶液,采用LC-MS/MS方法测定体系中产物甲羟戊酸的量,计算抑制率,得到洛伐他汀酸浓度-抑制率曲线。将血脂康样品溶液加入到酶促反应体系中,根据抑制率计算血脂康活性成分的含量。结果:洛伐他汀酸浓度为10~500 ng·m L-1范围内抑制效应显著,质量控制样品的准确度和精密度范围分别为95.28%~101.53%,7.08%~10.51%;平均加样回收率为100.25%,RSD为2.27%。检测得到3批血脂康胶囊活性成分的含量以洛伐他汀酸计分别为每粒4.86,4.71和4.56 mg。结论:建立的方法精密度好,准确度高,可用于血脂康胶囊的生物活性测定。酶法测定结果显著高于HPLC法的测得值,推测血脂康中可能有其他成份在同时发挥抑制HMG-Co A还原酶的作用。本研究结果对血脂康的质量控制具有指导意义。
[Abstract]:Objective: to establish a method for determining the activity of Xuezhikang. Methods: the enzymatic reaction of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase (HMG-Co reductase) was established. The amount of HMG-Co A reductase was 3.125 脳 10 ~ (-4) U and the amount of nicotinamide adenine dinucleotide phosphate was 160 nmol 路L ~ (-1). The amount of hydroxymethyl glutaric acid monoacyl-CoA HMG-Co A was 5 nmol 路L ~ (-1) and the reaction time was 50 min. A series of working solution of lovastatin acid was added. The concentration and inhibition rate curve of lovastatin acid was obtained by LC-MS/MS method. The sample solution of Xuezhikang was added to the enzymatic reaction system. The content of Xuezhikang was calculated according to the inhibition rate. Results: the inhibitory effect of lovastatin in the range of 10500 ng 路mL ~ (-1) was significant. The accuracy and precision range of the quality control samples were 95.281.53 and 10.51, respectively. The average recovery was 100.25% and RSD was 2.27.The three batches of active components of Xuezhikang capsules were determined to be 4.86 per lovastatin acid. Conclusion: the established method has good precision and high accuracy, and can be used for the determination of biological activity of Xuezhikang capsule. The result of enzymatic assay is significantly higher than that of HPLC method. It is speculated that other components in Xuezhikang may play a role in inhibiting HMG-Co A reductase at the same time. The results of this study are instructive to the quality control of Xuezhikang.
【作者单位】：烟台大学药学院;绿叶制药集团有限公司长效和靶向制剂国家重点实验室;北大维信生物技术有限公司;
【基金】：国家“重大新药创制”科技重大专项(2013ZX09402201)
【分类号】：R927.2;O657.63
【正文快照】： 他汀类药物通过竞争性地抑制胆固醇合成途径的限速酶HMG-Co A还原酶的活性,阻断胆固醇的生物合成,使血液中总胆固醇及低密度脂蛋白胆固醇含量下降,是一类临床广泛应用的降脂药[1]。已上市的该类药物有洛伐他汀、辛伐他汀和阿托伐他汀等[2]。血脂康胶囊(以下简称“血脂康”)是

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