Lenière Anne-Charlotte, Upadhyay Amit, Follet Jérôme, O'Sullivan Timothy P
University of Lille, CNRS, Centrale Lille, Junia, Université Polytechnique Hauts de France, UMR 8520, IEMN Institut d'Electronique de Microélectronique et de Nanotechnologie, F59000, Lille, France.
School of Chemistry, University College Cork, Cork, T12 YN60, Ireland; School of Pharmacy, University College Cork, Cork, T12 YN60, Ireland; Analytical and Biological Chemistry Research Facility, University College Cork, Cork, T12 YN60, Ireland.
Int J Parasitol Drugs Drug Resist. 2025 Apr 15;28:100592. doi: 10.1016/j.ijpddr.2025.100592.
The protozoan parasite Cryptosporidium is the etiological agent of cryptosporidiosis, a ubiquitous diarrheic disease affecting humans and animals. Treatment options are limited, highlighting an urgent need for novel therapeutics. Despite decades of research and a wide diversity of strategies to tackle parasite metabolic pathways, no completely effective drug has been identified to date. Within targeted parasite enzymatic and metabolic pathways, the synthesis of nucleotide mediated by the inosine 5'-monophosphate dehydrogenase (IMPDH) enzyme is the focus of significant research efforts. Based on our prior studies of bacterial IMPDH inhibitors, we report herein the development and characterisation of novel inhibitors targeting Cryptosporidium parvum IMPDH (CpIMPDH). Specifically, we synthesised heteroaryl-containing urea and squaramide analogues to evaluate their potential in vitro anti-Cryptosporidium activity. Initial screening identified nine active compounds with the most potent candidates achieving IC values as low as 2.2 μM. Subsequent time-course experiments revealed that the molecules effectively inhibit parasite invasion and early intracellular development but failed to tackle C. parvum growth when introduced at 30 h post infection. The present work introduces a new family of squaramide-derived IMPDH inhibitors and also interrogates the need to standardise commonly accepted protocols used for assessing anti-cryptosporidial drug activity.
原生动物寄生虫隐孢子虫是隐孢子虫病的病原体，这是一种影响人类和动物的普遍存在的腹泻疾病。治疗选择有限，凸显了对新型疗法的迫切需求。尽管经过数十年的研究以及针对寄生虫代谢途径的多种策略，但迄今为止尚未确定完全有效的药物。在靶向的寄生虫酶促和代谢途径中，由肌苷5'-单磷酸脱氢酶（IMPDH）介导的核苷酸合成是大量研究工作的重点。基于我们之前对细菌IMPDH抑制剂的研究，我们在此报告针对微小隐孢子虫IMPDH（CpIMPDH）的新型抑制剂的开发和表征。具体而言，我们合成了含杂芳基的脲和方酸二酰胺类似物，以评估它们在体外的抗隐孢子虫活性潜力。初步筛选确定了9种活性化合物，其中最有效的候选物的IC值低至2.2 μM。随后的时间进程实验表明，这些分子有效抑制寄生虫入侵和早期细胞内发育，但在感染后30小时引入时未能抑制微小隐孢子虫的生长。本研究介绍了一个新的方酸二酰胺衍生的IMPDH抑制剂家族，并探讨了标准化用于评估抗隐孢子虫药物活性的常用协议的必要性。
