National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido, Japan.
Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Dakahlia, Egypt.
Parasit Vectors. 2022 Sep 19;15(1):329. doi: 10.1186/s13071-022-05430-4.
An innovative approach has been introduced for identifying and developing novel potent and safe anti-Babesia and anti-Theileria agents for the control of animal piroplasmosis. In the present study, we evaluated the inhibitory effects of Malaria Box (MBox) compounds (n = 8) against the growth of Babesia microti in mice and conducted bioinformatics analysis between the selected hits and the currently used antibabesial drugs, with far-reaching implications for potent combinations.
A fluorescence assay was used to evaluate the in vivo inhibitory effects of the selected compounds. Bioinformatics analysis was conducted using hierarchical clustering, distance matrix and molecular weight correlation, and PubChem fingerprint. The compounds with in vivo potential efficacy were selected to search for their target in the piroplasm parasites using quantitative PCR (qPCR).
Screening the MBox against the in vivo growth of the B. microti parasite enabled the discovery of potent new antipiroplasm drugs, including MMV396693 and MMV665875. Interestingly, statistically significant (P < 0.05) downregulation of cysteine protease mRNA levels was observed in MMV665875-treated Theileria equi in vitro culture in comparison with untreated cultures. MMV396693/clofazimine and MMV665875/atovaquone (AV) showed maximum structural similarity (MSS) with each other. The distance matrix results indicate promising antibabesial efficacy of combination therapies consisting of either MMV665875 and AV or MMV396693 and imidocarb dipropionate (ID).
Inhibitory and hematology assay results suggest that MMV396693 and MMV665875 are potent antipiroplasm monotherapies. The structural similarity results indicate that MMV665875 and MMV396693 have a similar mode of action as AV and ID, respectively. Our findings demonstrated that MBox compounds provide a promising lead for the development of new antibabesial therapeutic alternatives.
为了控制动物巴贝虫病，我们引入了一种创新方法来鉴定和开发新型有效且安全的抗巴贝西虫和抗泰勒虫药物。在本研究中，我们评估了 Malaria Box (MBox) 化合物（n=8）对小鼠微小巴贝虫生长的抑制作用，并对选定的化合物与目前使用的抗巴贝斯虫药物进行了生物信息学分析，这对有效的组合具有深远的意义。
采用荧光测定法评估选定化合物的体内抑制作用。使用层次聚类、距离矩阵和分子量相关性以及 PubChem 指纹图谱进行生物信息学分析。选择具有体内潜在疗效的化合物，使用定量 PCR (qPCR) 搜索其在巴贝斯虫寄生虫中的靶点。
筛选 MBox 化合物对微小巴贝虫体内生长的抑制作用，发现了新型有效的抗巴贝斯虫新药，包括 MMV396693 和 MMV665875。有趣的是，与未处理的培养物相比，在 MMV665875 处理的体外培养的马泰勒虫中，半胱氨酸蛋白酶 mRNA 水平呈统计学显著（P<0.05）下调。MMV396693/氯法齐明和 MMV665875/阿托伐醌（AV）彼此之间具有最大的结构相似性（MSS）。距离矩阵结果表明，由 MMV665875 和 AV 或 MMV396693 和咪唑苯并二氮䓬（ID）组成的联合疗法具有有希望的抗巴贝斯虫疗效。
抑制和血液学检测结果表明，MMV396693 和 MMV665875 是有效的抗巴贝斯虫单药治疗药物。结构相似性结果表明，MMV665875 和 MMV396693 的作用模式分别与 AV 和 ID 相似。我们的研究结果表明，MBox 化合物为开发新型抗巴贝斯虫治疗替代品提供了有希望的先导化合物。
