Department of Biological Science, School of Pharmacy, São Paulo State University (UNESP), Araraquara, SP, Brazil.
Department of Biological Science, Institute of Bioscience, Letters and Exact Science, São Paulo State University (UNESP), Araraquara, SP, Brazil.
Protein Pept Lett. 2024;31(7):532-543. doi: 10.2174/0109298665308871240703090408.
Peptide drugs are advantageous because they are subject to rational design and exhibit highly diverse structures and broad biological activities. The NS2B-NS3 protein is a particularly promising flavivirus therapeutic target, with extensive research on the development of inhibitors as therapeutic candidates, and was used as a model in this work to determine the mechanism by which GA-Hecate inhibits ZIKV replication.
The present study aimed to evaluate the potential of GA-Hecate, a new antiviral developed by our group, against the Brazilian Zika virus and to evaluate the mechanism of action of this compound on the flavivirus NS2B-NS3 protein.
Solid-phase peptide Synthesis, High-Performance Liquid Chromatography, and Mass Spectrometry were used to obtain, purify, and characterize the synthesized compound. Real-time and enzymatic assays were used to determine the antiviral potential of GA-Hecate against ZIKV.
The RT-qPCR results showed that GA-Hecate decreased the number of ZIKV RNA copies in the virucidal, pre-treatment, and post-entry assays, with 5- to 6-fold fewer RNA copies at the higher nontoxic concentration in Vero cells (HNTC: 10 μM) than in the control cells. Enzymatic and kinetic assays indicated that GA-Hecate acts as a competitive ZIKV NS2B-NS3 protease inhibitor with an IC of 32 nM and has activity against the yellow fever virus protease.
The results highlight the antiviral potential of the GA-Hecate bioconjugate and open the door for the development of new antivirals.
肽类药物具有优势，因为它们可以进行合理设计，表现出高度多样化的结构和广泛的生物活性。NS2B-NS3 蛋白是一种特别有前途的黄病毒治疗靶点，广泛研究了抑制剂作为治疗候选物的开发，并在这项工作中被用作模型，以确定 GA-Hecate 抑制 ZIKV 复制的机制。
本研究旨在评估我们小组开发的新型抗病毒药物 GA-Hecate 对巴西寨卡病毒的潜力，并评估该化合物对黄病毒 NS2B-NS3 蛋白作用的机制。
采用固相肽合成、高效液相色谱和质谱法获得、纯化和表征合成化合物。实时和酶测定法用于确定 GA-Hecate 对 ZIKV 的抗病毒潜力。
RT-qPCR 结果表明，GA-Hecate 在溶病毒、预处理和进入后试验中减少了 ZIKV RNA 拷贝数，在更高的无毒浓度（HNTC：10 μM）下，Vero 细胞中的 RNA 拷贝数比对照细胞少 5-6 倍。酶和动力学测定表明，GA-Hecate 是一种竞争性 ZIKV NS2B-NS3 蛋白酶抑制剂，IC 为 32 nM，对黄热病毒蛋白酶具有活性。
这些结果突出了 GA-Hecate 生物缀合物的抗病毒潜力，并为开发新的抗病毒药物开辟了道路。
