Bora Kushal, Sarma Manash, Kanaujia Shankar Prasad, Dubey Vikash Kumar
School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi 221005, Uttar Pradesh, India; Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati - 781039, Assam, India.
School of Biochemical Engineering, Indian Institute of Technology (BHU), Varanasi 221005, Uttar Pradesh, India.
Free Radic Biol Med. 2024 Nov 20;225:275-285. doi: 10.1016/j.freeradbiomed.2024.10.257. Epub 2024 Oct 10.
The dual-target inhibitors (ZINC000008876351 and ZINC000253403245) were identified by utilizing an advanced computational drug discovery method by targeting two critical enzymes such as FeSODA (Iron superoxide dismutase) and TryR (Trypanothione reductase) within the antioxidant defense system of Leishmania donovani (Ld). In vitro enzyme inhibition kinetics reveals that both the compound's ability to inhibit the function of enzyme LdFeSODA and LdTryR with inhibition constant (Ki) value in the low μM range. Flow cytometry analysis, specifically at IC and 2X IC doses of both the compounds, the intracellular ROS was significantly increased as compared to the untreated control. The compounds ZINC000253403245 and ZINC000008876351 exhibited strong anti-leishmanial activity in a dose-dependent manner against both the promastigote and amastigote stages of the parasite. The data indicate that these molecules hold promise as potential anti-leishmanial agents for developing new treatments against visceral leishmaniasis, specifically targeting the LdFeSODA and LdTryR enzymes. Additionally, the in vitro MTT assay shows that combining these compounds with miltefosine produces a synergistic effect compared to miltefosine alone. This suggests that the compounds can boost miltefosine's effectiveness by synergistically inhibiting the growth of L. donovani promastigotes. Given the emergence of miltefosine resistance in some Leishmania strains, these findings are particularly significant.
通过利用先进的计算药物发现方法，针对杜氏利什曼原虫（Ld）抗氧化防御系统中的两种关键酶，如铁超氧化物歧化酶（FeSODA）和锥虫硫醇还原酶（TryR），鉴定出了双靶点抑制剂（ZINC000008876351和ZINC000253403245）。体外酶抑制动力学表明，这两种化合物都有能力抑制LdFeSODA和LdTryR酶的功能，其抑制常数（Ki）值处于低 microM 范围。流式细胞术分析显示，特别是在两种化合物的IC和2X IC剂量下，与未处理的对照相比，细胞内活性氧显著增加。化合物ZINC000253403245和ZINC000008876351对寄生虫的前鞭毛体和无鞭毛体阶段均表现出强烈的剂量依赖性抗利什曼原虫活性。数据表明，这些分子有望成为开发针对内脏利什曼病新疗法的潜在抗利什曼原虫药物，特别是针对LdFeSODA和LdTryR酶。此外，体外MTT试验表明，与单独使用米替福新相比，将这些化合物与米替福新联合使用会产生协同效应。这表明这些化合物可以通过协同抑制杜氏利什曼原虫前鞭毛体的生长来提高米替福新的有效性。鉴于一些利什曼原虫菌株出现了对米替福新的耐药性，这些发现尤为重要。
