基于药物代谢酶调控的参芎葡萄糖注射液PK-PD相关性及配伍机制研究

Shenxiong glucose injection (SGI) is one of the main drugs for prevention and treatment of cardiovascular and cerebrovascular diseases in China. It has remarkable clinical effects and has been on the market for many years. In previous studies, we found that the pharmacodynamics and pharmacokinetic parameters were changed significantly in rats, and the activity of CYP450 enzyme had different effects in mice, which were detected before and after salvia miltiorrhiza combined with ligustrazine. In view of the pharmacokinetic study of SGI in disease states and the influence of drug metabolism enzymatic activity induced by the active ingredient of SGI were not reported. In addition, the compatibility mechanism is not clear. On the basis of preliminary research, the process of distribution, metabolism and excretion of main active components (salvianic acid, purple oxalic acid, danphenolic acid D, rosemary acid, and danphenate B) in Salvia miltiorrhiza , ligustrazine or formula SGI will be carried out respectively in myocardial ischemia model rats. Meanwhile, Study on the relationship between pharmacokinetic process and the change of pharmacological effect (PK-PD). Furthermore, from the perspective of drug metabolism enzyme regulation, study the expression of I, II phase metabolism enzyme and the conversion process of active constituents and its metabolites in vivo. It will elucidate the compatibility mechanism and safety evaluation of SGI to guide clinical treatment.

参芎葡萄糖注射液（SGI）作为我国防治心脑血管疾病的主要药物之一，已上市销售多年且临床疗效显著。课题组在前期研究中发现丹参和川芎嗪在配伍前后大鼠体内的药效及药动学参数均发生了明显变化，且对小鼠CYP450酶活性有不同程度的影响。鉴于SGI在病理状态下的药动学研究及组方中各效应成分对药物代谢酶活性的影响未见报道,且配伍机制尚不明确。本项目拟在前期研究的基础上，研究丹参、川芎嗪及其组方SGI中主要活性成分（丹参素、紫草酸、丹酚酸D、迷迭香酸、丹酚酸B、川芎嗪等）在心肌缺血模型大鼠体内的分布、代谢、排泄过程，及其与药理效应消长变化之间的关系（PK-PD相关性），并从药物代谢酶调控的角度，研究I相、II相代谢酶的表达及其介导的活性成分及代谢产物的体内转化过程，为阐明中药配伍及安全性评价提供理论依据,有助于指导临床合理用药。

参芎葡萄糖注射液由丹参和川芎嗪配伍组成，主要用于缺血性血管疾病和闭塞性脑血管病的质量，已上市销售多年并广泛应用于临床。但其在病理状态下配伍前后各效应成分对药物代谢酶表达的影响及体内过程研究尚属空白，无法从在体的角度去阐释配伍机制。本课题研究发现配伍前后川芎嗪的药代动力学参数有明显改变，这可能是由于配伍后丹参改变了介导川芎嗪Ⅰ 相代谢的CYP1A2、2C9、3A4的活性和表达，单次给药时CYP1A2、CYP2C11、CYP3A4三种酶的表达增加，推测活性增加，使得川芎嗪的代谢加快，川芎嗪的体内吸收总量降低；多次给药则相反。在组织分布研究中，我们发现配伍后（单次给药）情况下能显著提高靶器官（心）及组织中的丹参素和川芎嗪的药物浓度，且在配伍后含量最高的组织由配伍前的代谢器官（肾）变为靶器官（心）。而PK-PD研究提示配伍后AST、CK-MB、cTn-I 3个药效指标最大值与血药浓度的达峰时间相同，不存在滞后现象。配伍后川芎嗪、丹参素作用于AST、CK-MB、cTn-I的活性优于配伍前，这与SGI的PK-PD多效应成分整合结果一致。这说明在首次给药时，活性成分在配伍的情况下可迅速分布至靶器官发挥药效，而在血中含量较低，在多次给药后，体内的吸收逐渐增加，血液中的活性成分进入心肌、血管平滑肌等效应部位的速度和药物量增加，直接在效应部位发挥药效。排泄和代谢研究结果显示，各成分在组织中不存在长时间蓄积现象，丹参素可经过原形主要经肾排出体外（累积排泄率32~33%），迷迭香酸和川芎嗪经尿液、粪便及胆汁的排泄量均很少，累积排泄率均不足1%。在大鼠尿液、粪便、胆汁中共鉴定出 2 个原形成分和26个代谢产物，其中25个为丹酚酸类成分的代谢产物，1个为川芎嗪的代谢产物。配伍后丹参类成分的代谢产物的数量和峰面积大于单方组，基于组方药效优于单方的结论，推测代谢产物中可能存在活性代谢产物。综上，通过代谢酶介导的药动行为改变机制、PK-PD及体内过程研究，初步阐释了SGI配伍的合理性和优势。