二价HIV-1融合抑制剂的设计、合成与活性评价

针对人免疫缺陷病毒跨膜糖蛋白(HIV-1 gp41)N末端重复序列靶标设计二价融合抑制剂, 以C肽为模板, 通过共价交联形成类似发夹结构的相互平行的2条肽链, 研究了二价C肽分子不同连接位点与不同连接臂对抗HIV融合活性的影响. 细胞-细胞融合活性测试表明, 与单价分子相比, 所设计的基于N末端交联的C34或T20的二价分子在前体共价交联后, 活性明显提升. 基于C34或T20的N末端与C末端均存在发生协同效应的可能性, 在C34的N末端设计中β-丙氨酸为最适连接臂, 而在C末端的设计中C34C的融合活性提高最大. 单价分子CβAC34经过氧化形成二硫键连接的二价分子BiCβAC34, 融合活性从43.7 nmol/L提高到6.4 nmol/L, 表明二价抑制剂中2条C肽链间具有良好的协同效应. 本文结果表明, 针对gp41靶标设计的二价融合抑制剂能够相互协同.

Design,Synthesis and Activity Evaluation of Novel Bivalent HIV-1 Fusion Inhibitor

Multivalent inhibitor is an efficient strategy for inhibitor design based on an additive effect of ΔG. In this work, we chose C34 and T20 as the template sequences to study bivalent inhibitors targeting HIV-1 gp41 NHR domain. We optimized the crosslink sites and linkers to improve anti-HIV activities of the bivalent fusion inhibitors. Compared to monovalent molecules, significant cooperative effects in the anti-cell-cell fusion activity were observed in the bivalent inhibitors, at either N- or C-terminal crosslink of both C34 and T20. β-Alanine may be the most suitable linker for N-terminal crosslink, while C34C is the best C-terminal crosslinked molecule. Specially, the anti-HIV IC₅₀ value of peptide BiCβAC34 was improved from 43.7 nmol/L to 6.4 nmol/L, indicating the two C-peptide chains had a cooperative effect. The results show that bivalent fusion inhibitor for gp41 design could appear a cooperative effect.