Laboratório Nacional de Biociências (LNBio), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP 13083-970, Brazil.
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP 05508-900, Brazil.
Molecules. 2020 May 30;25(11):2551. doi: 10.3390/molecules25112551.
High genetic and phenotypic variability between species and strains within species make the development of broad-spectrum antileishmanial drugs challenging. Thus, screening panels consisting of several diverse species can be useful in enabling compound prioritization based on their spectrum of activity. In this study, a robust and reproducible high content assay was developed, and 1280 small molecules were simultaneously screened against clinically relevant cutaneous and visceral species: , , and . The assay is based on THP-1 macrophages infected with stationary phase promastigotes and posterior evaluation of both compound antileishmanial activity and host cell toxicity. The profile of compound activity was species-specific, and out of 51 active compounds, only 14 presented broad-spectrum activity against the three species, with activities ranging from 52% to 100%. Notably, the compounds CB1954, Clomipramine, Maprotiline, Protriptyline, and ML-9 presented pan-leishmanial activity, with efficacy greater than 70%. The results highlight the reduced number of compound classes with pan-leishmanial activity that might be available from diversity libraries, emphasizing the need to screen active compounds against a panel of species and strains. The assay reported here can be adapted to virtually any species without the need for genetic modification of parasites, providing the basis for the discovery of broad spectrum anti-leishmanial agents.
种间和种内菌株间的高遗传和表型变异性使得开发广谱抗利什曼原虫药物具有挑战性。因此，由几种不同物种组成的筛选面板可用于根据其活性谱对化合物进行优先级排序。在这项研究中，开发了一种稳健且可重复的高内涵测定法，并对 1280 种小分子同时进行了针对临床相关皮肤和内脏物种的筛选： 、 、 和 。该测定法基于用静止期前鞭毛体感染 THP-1 巨噬细胞，随后评估化合物的抗利什曼原虫活性和宿主细胞毒性。化合物活性谱具有物种特异性，在 51 种活性化合物中，只有 14 种对三种物种具有广谱活性，活性范围为 52%至 100%。值得注意的是，化合物 CB1954、氯米帕明、马普替林、普罗替林和 ML-9 具有泛利什曼原虫活性，疗效大于 70%。结果表明，多样性文库中可能具有泛利什曼原虫活性的化合物类别的数量减少，这强调了需要针对一组物种和菌株筛选活性化合物。这里报道的测定法可以适用于几乎任何物种，而无需对寄生虫进行遗传修饰，为发现广谱抗利什曼原虫药物提供了基础。
