Department of Anesthesiology and Perioperative Medicine, Penn State College of Medicine, Hershey, Pennsylvania, USA.
Physiol Rep. 2023 Jul;11(14):e15760. doi: 10.14814/phy2.15760.
All currently employed pharmaceutical formulations of hydroxychloroquine (HCQ) sulfate are a racemate, consisting of equal parts mixture of two stereoisomers: R(-)HCQ and S(+)HCQ sulfates. The aims of the current study were first, to obtain and characterize pure HCQ enantiomers. The separation and purification of free base HCQ enantiomers from the racemate form were performed using semi-preparative chiral high-performance liquid chromatography. Second, we compared the pharmacological properties of both optical isomers and racemic mixture on the intracellular Ca oscillations employing an in vitro model of human cardiomyocytes derived from induced pluripotent stem cells (iPSCs). The results of the pharmacological investigations indicate that the racemic and pure stereoisomer forms of HCQ sulfate exhibited a dose-dependent inhibition of spontaneous Ca oscillations (as measured from their frequency and Ca peak widths) in cardiomyocytes 5-45 min following exposure. In addition, the concentration-response relationships for all three compounds indicated that the rank order of potency (IC ) was R(-)HCQ >racemic HCQ >S(+)HCQ for the frequency of the Ca oscillations and width of Ca peaks for all time points examined. These studies indicate that both R(-) and S(+) stereoisomers exhibit differing pharmacological actions on hiPSC cardiomyocytes, with the former effecting a greater potency on cell Ca handling.
所有目前市售的羟氯喹硫酸盐制剂均为外消旋体，由两种立体异构体的等量混合物组成：R(-)HCQ 和 S(+)HCQ 硫酸盐。本研究的目的首先是获得并表征纯 HCQ 对映异构体。从外消旋形式中分离和纯化游离碱 HCQ 对映异构体是通过半制备手性高效液相色谱来完成的。其次，我们比较了两种光学异构体和外消旋混合物在人诱导多能干细胞 (iPSC) 衍生的心肌细胞内细胞内 Ca 振荡上的药理学特性。药理学研究的结果表明，HCQ 硫酸盐的外消旋和纯立体异构体形式在暴露后 5-45 分钟内以剂量依赖性方式抑制心肌细胞中的自发 Ca 振荡（从其频率和 Ca 峰宽来测量）。此外，所有三种化合物的浓度-反应关系表明，所有检测时间点的 Ca 振荡频率和 Ca 峰宽的效力（IC ）顺序为 R(-)HCQ >外消旋 HCQ >S(+)HCQ。这些研究表明，两种 R(-) 和 S(+) 立体异构体对 hiPSC 心肌细胞表现出不同的药理学作用，前者对细胞 Ca 处理具有更大的效力。
