The molecular target of bicyclams, potent inhibitors of human immunodeficiency virus replication
The molecular target of bicyclams, potent inhibitors of human immunodeficiency virus replication
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DOI：
10.1128/jvi.70.2.689-696.1996
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发表时间：
1996-02-01
影响因子：
5.4
通讯作者：
Werner, G
中科院分区：
文献类型：
--
作者：
DeVreese, K;KoflerMongold, V;Werner, G
双环胺是一类新型的抗病毒化合物，其作为人免疫缺陷病毒1型（HIV-1）和HIV-2复制的有效和选择性抑制剂。它们在吸附到CD 4受体和逆转录之前阻断病毒生命周期的早期步骤，为了鉴定这些化合物的分子靶点，我们对HIV-1分子克隆NL 4 -3的变体进行了遗传分析，该变体对两种结构相关的双环类药物JM 2763和更有效的SID 791产生了抗性。在存在逐渐增加的化合物浓度的情况下在MT-4细胞中长期传代后获得抗性菌株，通过使标记拯救技术适应MT-4细胞来产生抗性菌株的选定基因与亲本NL 4 -3前病毒之间的互补物，通过将抗双环胺病毒的包膜gp 120基因转移到NL 4 -3亲本遗传背景中来拯救双环胺抗性表型，在耐药菌株的gp 120基因中，我们鉴定了导致V3环氨基酸取代的几个突变。此外，在SID 791和JM 2763耐药菌株中均发现了V3和V4环二硫键附近的两个高度保守的氨基酸取代。V5和C5存在于SID 791抗性病毒中，用包膜基因的重叠部分进行的纯化实验表明，如果不是全部的话，大多数突变是发展完全的SID 791抗性表型所必需的，C基因的突变-V3环序列下游的gp 120末端部分赋予对JM 2763的部分抗性，但没有显著降低对JM 2763的敏感性。SID 791抗HIV病毒的遗传数据和生物学特性表明抑制进入和融合是HIV抑制双环类化合物的作用方式。本文还讨论了双环类化合物与gp 120结合抑制gp 120解折叠和从gp 41融合结构域脱落的可能机制。
;/Bicyclams are a novel class of antiviral compounds which act as potent and selective inhibitors of the replication of human immunodeficiency virus type 1 (HIV-1) and HIV-2. They block an early step in the viral life cycle following adsorption to the CD4 receptor and preceding reverse transcription, To identify the molecular target of these compounds, we genetically analyzed variants of the HIV-1 molecular clone NL4-3, which developed resistance against two structurally related bicyclams, JM2763 and the more potent SID791. The resistant strains were obtained after long-term passaging in MT-4 cells in the presence of progressively increasing compound concentrations, Recombinants between selected genes of the resistant strains and the parental NL4-3 provirus were generated by adapting the marker rescue technique to MT-4 cells, The bicyclam-resistant phenotype was rescued by transferring the envelope gp120 gene of bicyclam-resistant virus into the NL4-3 parental genetic background, In the gp120 genes of the resistant strains, we identified several mutations leading to amino acid substitutions in the V3 loop, Furthermore, two substitutions of highly conserved amino acids in close proximity to the disulfide bridges of the V3 and V4 loops were found in both SID791- and JM2763-resistant strains, Additional mutations in regions encoding V3, C4, V5, and C5 were present in SID791-resistant viruses, Recombination experiments with overlapping parts of the envelope gene indicated that most, if not all, of the mutations were necessary to develop the fully SID791 resistant phenotype, The mutations in the C-terminal part of gp120 downstream of the V3 loop sequence conferred partial resistance to JM2763 but did not significantly decrease susceptibility to SID791. The genetic data and the biological properties of the resistant viruses point to inhibition of entry and fusion as the mode of action of the HIV-inhibitory bicyclams, A possible mechanism of binding of bicyclams to gp120 leading to inhibition of unfolding of gp120 and its shedding from the gp41 fusion domain is discussed.