Department of Biotechnology and Bioengineering, Chonnam National University, 77 Yongbong-ro, Buk-gu, Gwangju 500-757, Korea.
Molecules. 2013 Dec 13;18(12):15600-12. doi: 10.3390/molecules181215600.
The discovery of potent therapeutic compounds against dengue virus is urgently needed. The NS2B-NS3 protease (NS2B-NS3pro) of dengue fever virus carries out all enzymatic activities needed for polyprotein processing and is considered to be amenable to antiviral inhibition by analogy. Virtual screening of 300,000 compounds using Autodock 3 on the GVSS platform was conducted to identify novel inhibitors against the NS2B-NS3pro. Thirty-six compounds were selected for in vitro assay against NS2B-NS3pro expressed in Pichia pastoris. Seven novel compounds were identified as inhibitors with IC50 values of 3.9 ± 0.6-86.7 ± 3.6 μM. Three strong NS2B-NS3pro inhibitors were further confirmed as competitive inhibitors with Ki values of 4.0 ± 0.4, 4.9 ± 0.3, and 3.4 ± 0.1 μM, respectively. Hydrophobic and hydrogen bond interactions between amino acid residues in the NS3pro active site with inhibition compounds were also identified.
急需发现针对登革热病毒的有效治疗化合物。登革热病毒的 NS2B-NS3 蛋白酶（NS2B-NS3pro）进行多蛋白加工所需的所有酶活性，并且可以通过类比来抑制抗病毒。使用 GVSS 平台上的 Autodock 3 对 300,000 种化合物进行虚拟筛选，以鉴定针对 NS2B-NS3pro 的新型抑制剂。选择了 36 种化合物用于针对在巴斯德毕赤酵母中表达的 NS2B-NS3pro 的体外测定。鉴定出 7 种新型化合物为具有 3.9 ± 0.6-86.7 ± 3.6 μM 的 IC50 值的抑制剂。进一步确认了 3 种强 NS2B-NS3pro 抑制剂为竞争性抑制剂，其 Ki 值分别为 4.0 ± 0.4、4.9 ± 0.3 和 3.4 ± 0.1 μM。还确定了 NS3pro 活性部位的氨基酸残基与抑制化合物之间的疏水和氢键相互作用。
