华法林个体化用药的血浆药物代谢组学研究及机制探讨

Recently，individualization of warfarin stable dosage is commonly researched. Genetic polymorphisms in vitamin K epoxide reductase complex 1 (VKORC1) and cytochrome P450 2C9 (CYP2C9) and age, gender and body weight which had attracted broad attention could account for 30% to 60% of the dose variation. In recent years, ‘Pharmaco-metabolomics’ which affected by gene, environment and physiological/ pathological functions could predict the outcome of a drug or xenobiotic intervention in an individual based on a mathematical model of pre-intervention metabolite signatures. In our previous research, the metabolome of patients with valvular heart disease were associated to INR. In this study, Patients with valvular heart disease are recruited, and the pharmaco-metabolomics （GC-MS/UPLC-Q-TOFMS）, pharmacogenomics and PCA/PLSDA approach is used to predict the interindividual variability of steady-state dose and responsivity of warfarin. Metabolomics in preoperative and postoperative baseline plasma is performed on patients with valvular heart disease. Correlation analysis is conducted with data relating to predose plasma metabolites to predict the steady-state trough concentration and select the metabolites that substantially contributed to such prediction. The selection of these metabolites are to generate mathematical models for clinical prediction. The molecular mechanisms of pharmacodynamics-Pharmacometabolomics individual variation are explored using metabolic network analysis of these markers after validated in clinically. This research is of great theoretical and practical significance to facilitate individualized drug therapy and provide in-depth understanding of individual variation of warfarin. Pharmaco-metabolomics platform of personalized medicine is of course worth waiting for based on this study.

华法林稳态剂量的个体化是近年来广泛研究的课题。然而，目前结合基因、年龄、性别等因素的剂量预测模型，仅能解释约30-60%的个体差异，尚不能满足临床个体化用药的需求。受遗传、环境及生理病理综合影响的药物代谢组学可根据基础代谢谱的差异对药物反应性进行预测。前期研究发现心脏瓣膜置换术患者的代谢组与INR之间存在一定的相关性。本项目拟以心脏瓣膜置换术患者为研究对象，运用基于质谱法的药物代谢组学方法、结合药物基因组学及主成分分析、判别分析等研究手段表征机体个体差异并探讨不同个体内源性代谢特征与华法林应答的规律，通过测定患者术前/术后血浆代谢谱，预测华法林的稳态给药剂量，提取与药物敏感性相关的生物标记物；经临床可靠性和普适性验证后通过代谢通路分析探索华法林药物敏感性差异的分子机制。本课题旨在为实现临床华法林个体化合理用药提供技术支撑，并最终以此为基础建立药物代谢组学个体化研究平台。

华法林稳态剂量的个体化是近年来广泛研究的课题。然而，目前结合基因、年龄、性别等因素的剂量预测模型，仅能解释约30-60%的个体差异，尚不能满足临床个体化用药的需求。受遗传、环境及生理病理综合影响的药物代谢组学可根据基础代谢谱的差异对药物反应性进行预测。本项目以心脏瓣膜置换术患者为研究对象，共入组296例患者，测定VKORC及CYP2C9的基因、稳定华法林使用剂量、INR，并收集相关临床资料，运用基于质谱法的药物代谢组学方法发现VKORC基因型为AA及GA患者之间的差异主要集中在精氨酸与脯氨酸代谢以及鞘脂代谢通路中；推测CYP2C9基因型为AA及GA患者之间的差异主要集中在精氨酸与脯氨酸代谢通路中，患者对华法林敏感与否可能与Phenylalanine metabolism和Linoleic acid metabolism有关，预测公式的准确与否可能与Arginine and proline metabolism和Steroid hormone biosynthesis有关。此外本课题将预测公式改为D=5.174-0.014*Age+0.009*Weight-0.0287*CYP2C9-2.108*VK1-3.438*VK2+0.00012*Ct238-0.4×Adr可将预测准确度提高3%，为实现临床华法林个体化合理用药提供技术支撑，多中心验证后可以向临床进一步推广。