亚砷酸毒性机制及预警研究

Arsenic trioxide (ATO) had dramatically empowered the most pernicious leukemia-acute promyelocytic leukemia (APL) to achieve the best treatment effects. It was independently researched and developed in China, and its appearance has a great impact on the international medical circles, arousing widely appreciated. However, ATO also has toxicities, the studies related predictive factors have been reported rarely..This project intends to make a fundamental research on the predictive factors about the toxicities, such as hyperleukocytosis, liver damage, caused by ATO. Firstly, focus on the metabolites of ATO: the abilities of inducing cell apoptosis are different among various arsenic metabolites, they are the determinant of toxicities. Previous studies were confined to the concentration of total arsenic. Unprecedentedly, we have first comprehensively analyzed the relationship among different arsenic metabolites with various toxicities. Secondly, draw attention to the body response to ATO, systematically explore the predictive factors on the toxic and side effects from multi-perspective, such as the enzymes which have great influence on the arsenic metabolism, the substances which are easily integrated with arsenic and so on. .This project will devote on the basic research for the predictive factors about the toxic and side effects of ATO, via Translational Medicine, establish toxic related predictive strategies, lay a foundation for ATO individual therapy both in APL and other diseases, make ATO, the independently researched and developed drug by Chinese, be safely used. Meanwhile, the related predictive strategies contain great value, worthy of promotion to the safety forecast on other drugs. .

亚砷酸注射液是我国自主研发的治疗急性早幼粒细胞白血病的特效药物，使该病从白血病中最凶险的病种改变为疗效最好的一型，在国际医药学界产生了巨大的影响，受到同行广泛认可。但亚砷酸具有一定毒性，对于其机制及预警研究鲜有报道。.拟对亚砷酸导致的高白细胞血症、肝脏损害等多种毒性的机制及预测因素进行研究。首先针对药物的代谢产物：各种砷代谢产物促细胞凋亡能力有很大差别，其浓度是决定毒性的重要因素，以往研究仅限于总砷浓度，我们率先综合分析各种砷代谢产物与多种毒性的关系。另一方面是机体对药物的反应，从影响砷代谢的酶、易与砷结合的物质等多个角度系统的研究毒性机制及预测因素。.本项目针对亚砷酸毒性的机制及预测因素进行基础研究，并将通过转化医学建立亚砷酸毒性预警策略，为应用亚砷酸个体化治疗急性早幼粒细胞白血病等疾病打下基础，使这一我国独立研发的药物安全的使用，并将部分方法推广到其他药物安全性预测研究中。

建立了HPLC-HG-AFS及HPLC-ICP-MS检测平台，首先检测到APL患者血浆中4种砷化物：三价无机砷（AsIII）、二甲基砷酸（DMAV）、一甲基砷酸（MMAV）和五价无机砷（AsV）。发现APL患者血浆中砷及其代谢产物的浓度特征：DMAV>AsIII、MMAV>AsV(p<0.05)，DMAV的含量较高，AsⅤ含量最低。血浆中的主要砷代谢产物为DMAV和MMAV。进行了APL患者红细胞中砷及其代谢产物：无机砷（iAs=AsV+AsIII）、DMAV、MMAV等含量研究，发现了红细胞中砷化物与血红蛋白结合，而储存于红细胞中。APL患者红细胞中的达稳态时，砷及其代谢产物的浓度特征为：iAs>MMA>DMA(p<0.05)。红细胞中MMA含量远高于DMA。红细胞中砷及其代谢产物浓度及比例与血浆中的均存在很大差异。APL患者用药达到稳态时，尿中砷及其代谢产物的总体浓度特征为：AsIII、DMAV>MMAV>AsV(p<0.0001)，主要的代谢产物为 DMAV和MMAV；AsIII原型和DMAV占比较高。尿中与血中砷形态浓度及占比存在正相关。.发现了As2O3（ATO）慢点法给药与常规法给药的药代动力参数的差异，证明了慢点法毒副性作用小、疗效更佳的原因，为初发APL的治疗提供了一种优秀的替代方案。急性肾功损伤需要接受连续肾替代治疗CVVHD的APL患者，对比CVVHD与非CVVHD下浓度特征、药时曲线及药动学参数比较：CVVHD增加了患者砷化物的清除，但随着药物持续输注，疗程内砷浓度能达到有效浓度以上，为临床个体化治疗提供依据。.分析了APL患者体内砷代谢产物水平及比例差异，结合SPN技术，首次证明了砷甲基转移酶（AS3MT）基因多态性通过调节砷代谢而影响亚砷酸治疗APL的疗效和毒副作用。发现AS3MT14215等基因型与砷甲基化代谢物具有相关性，治疗前检测AS3MT基因多态性情况，可提前预知患者肝损伤、高白细胞血症、DS等的发生，为临床提供毒副反应预警依据，以便采取措施加以预防，临床意义重大。.建立了单药ATO治疗APL早期死亡的风险分级系统，对早期死亡发生风险高的APL患者鉴别和治疗具有重要意义；探寻了巯基与ATO治疗的APL患者肝功能损伤相关性，巯基含量可预测发生肝功能损伤的程度。