Chakraborti Tapati, Das Partha, Choudhury Rajdeep, De Tripti
Indian J Biochem Biophys. 2015 Feb;52(1):14-22.
Proteases have been considered as an important group of targets for development of antiprotozoal drugs due to their essential roles in host-parasite interactions, parasite immune evasion, life cycle transition and pathogenesis of parasitic diseases. The development of potent and selective serine protease inhibitors targeting L. donovani secretory serine protease (pSP) could pave the way to the discovery of potential antileishmanial drugs. Here, we employed different classical serine protease inhibitors (SPIs), such as aprotinin, N-tosyl-1-phenylalanine chloromethyl ketone (TPCK), N-tosyl-lysine chloromethyl ketone (TLCK), benzamidine (Bza) and pSP-antibody to determine the role of the protease in parasitic survival, growth and infectivity. Among the different classical SPIs, aprotinin appeared to be more potent in arresting L. donovani promastigotes growth with significant morphological alterations. Furthermore, aprotinin and anti-pSP treated parasites significantly decreased the intracellular parasites and percentage of infected macrophages. These results suggest that SPIs may reduce the infectivity by targeting the serine protease activity and may prove useful to elucidate defined molecular mechanisms of pSP, as well as for the development of novel antileishmanial drugs in future.
由于蛋白酶在宿主 - 寄生虫相互作用、寄生虫免疫逃避、生命周期转变和寄生虫病发病机制中发挥着重要作用，它们一直被视为抗寄生虫药物开发的重要靶点类别。开发针对杜氏利什曼原虫分泌性丝氨酸蛋白酶（pSP）的强效且选择性的丝氨酸蛋白酶抑制剂，可能为发现潜在的抗利什曼原虫药物铺平道路。在此，我们使用了不同的经典丝氨酸蛋白酶抑制剂（SPIs），如抑肽酶、N - 对甲苯磺酰 - 1 - 苯丙氨酸氯甲基酮（TPCK）、N - 对甲苯磺酰 - 赖氨酸氯甲基酮（TLCK）、苯甲脒（Bza）和pSP抗体，以确定该蛋白酶在寄生虫存活、生长和感染性中的作用。在不同的经典SPIs中，抑肽酶似乎在抑制杜氏利什曼原虫前鞭毛体生长方面更有效，且伴有显著的形态学改变。此外，经抑肽酶和抗pSP处理的寄生虫显著减少了细胞内寄生虫数量以及感染巨噬细胞的百分比。这些结果表明，SPIs可能通过靶向丝氨酸蛋白酶活性来降低感染性，并且可能有助于阐明pSP的特定分子机制，以及在未来开发新型抗利什曼原虫药物。
