Xie Chunfang, Albulescu Laura-Oana, Bittenbinder Mátyás A, Somsen Govert W, Vonk Freek J, Casewell Nicholas R, Kool Jeroen
Amsterdam Institute of Molecular and Life Sciences, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Science, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081HV Amsterdam, The Netherlands.
Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands.
Biomedicines. 2020 Aug 20;8(9):297. doi: 10.3390/biomedicines8090297.
Animal-derived antivenoms are the only specific therapies currently available for the treatment of snake envenoming, but these products have a number of limitations associated with their efficacy, safety and affordability for use in tropical snakebite victims. Small molecule drugs and drug candidates are regarded as promising alternatives for filling the critical therapeutic gap between snake envenoming and effective treatment. In this study, by using an advanced analytical technique that combines chromatography, mass spectrometry and bioassaying, we investigated the effect of several small molecule inhibitors that target phospholipase A (varespladib) and snake venom metalloproteinase (marimastat, dimercaprol and DMPS) toxin families on inhibiting the activities of coagulopathic toxins found in snake venoms. The venoms of , , and , which are known for their potent haemotoxicities, were fractionated in high resolution onto 384-well plates using liquid chromatography followed by coagulopathic bioassaying of the obtained fractions. Bioassay activities were correlated to parallel recorded mass spectrometric and proteomics data to assign the venom toxins responsible for coagulopathic activity and assess which of these toxins could be neutralized by the inhibitors under investigation. Our results showed that the phospholipase A-inhibitor varespladib neutralized the vast majority of anticoagulation activities found across all of the tested snake venoms. Of the snake venom metalloproteinase inhibitors, marimastat demonstrated impressive neutralization of the procoagulation activities detected in all of the tested venoms, whereas dimercaprol and DMPS could only partially neutralize these activities at the doses tested. Our results provide additional support for the concept that combinations of small molecules, particularly the combination of varespladib with marimastat, serve as a drug-repurposing opportunity to develop new broad-spectrum inhibitor-based therapies for snakebite envenoming.
动物源抗蛇毒血清是目前治疗蛇咬伤的唯一特效疗法，但这些产品在疗效、安全性和可承受性方面存在一些局限性，难以用于热带地区的蛇咬伤受害者。小分子药物及候选药物被视为填补蛇咬伤与有效治疗之间关键治疗空白的有前景的替代方案。在本研究中，我们使用一种结合了色谱法、质谱法和生物测定法的先进分析技术，研究了几种靶向磷脂酶A（伐瑞拉地布）和蛇毒金属蛋白酶（马立马司他、二巯丙醇和二巯丙磺钠）毒素家族的小分子抑制剂对抑制蛇毒中促凝毒素活性的影响。以具有强大血液毒性而闻名的、、和的毒液，先用液相色谱法在高分辨率下分馏到384孔板上，然后对所得馏分进行促凝生物测定。将生物测定活性与同步记录的质谱和蛋白质组学数据相关联，以确定导致促凝活性的毒液毒素，并评估所研究的抑制剂能够中和哪些毒素。我们的结果表明，磷脂酶A抑制剂伐瑞拉地布中和了所有测试蛇毒中发现的绝大多数抗凝活性。在蛇毒金属蛋白酶抑制剂中，马立马司他对所有测试毒液中检测到的促凝活性表现出令人印象深刻的中和作用，而二巯丙醇和二巯丙磺钠在测试剂量下只能部分中和这些活性。我们的结果为以下概念提供了更多支持，即小分子组合，特别是伐瑞拉地布与马立马司他的组合，为开发基于新型广谱抑制剂的蛇咬伤治疗方法提供了药物重新利用的机会。
