Revisiting the mechanism of enfuvirtide and designing an analog with improved fusion inhibitory activity by targeting triple sites in gp41
Revisiting the mechanism of enfuvirtide and designing an analog with improved fusion inhibitory activity by targeting triple sites in gp41
复制标题
重新审视恩夫韦肽的机制并通过靶向 gp41 中的三重位点设计具有改进的融合抑制活性的类似物
DOI：
10.1097/qad.0000000000002208
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发表时间：
2019-08-01
期刊：
影响因子：
3.8
通讯作者：
Lu, Lu
中科院分区：
文献类型：
--
作者：
Xu, Wei;Pu, Jing;Lu, Lu
目的：为了重新审视恩夫韦肽（T20）的作用机制，并基于新定义的机制，设计具有改善的抗病毒活性的T20的类似肽。设计：我们比较了T20和T1144在共培养表达HIV-1包膜（Env）的中国仓鼠卵巢（CHO-Env）后不同时间对六螺旋束（6 HB）形成的抑制活性。细胞和表达CD 4（+）的MT-2细胞在31.5 ℃下的溶血作用，以及T20-SF（具有额外的富含Escherichan基序的T20的类似肽）对由FP-P（其含有融合肽和融合肽（FP）近端区（FPPR））介导的溶血作用和HIV-1感染的作用。在温控细胞-细胞融合试验中，使用6 HB特异性mAb 2G 8，通过流式细胞术检测肽对6 HB形成的抑制活性;通过p24和细胞-细胞融合试验评估肽对HIV-1感染和融合的抑制活性。结果：T20对HIV-1融合早期的6 HB形成有抑制作用，而T1144对融合晚期的6 HB形成无抑制作用。结论：T20-SF的N端和C端分别与N端七肽重复序列和FPPR结合，提示T20-SF具有双靶机制。基于这种新机制设计的T20-SF具有显著提高的抗HIV-1活性，因为它靶向gp 41中的三重位点，包括N-末端七肽重复序列、FPPR和融合肽。因此，本研究为设计具有改进的抗病毒活性的新型HIV融合抑制剂提供了线索。版权所有（C）2019威科医疗集团All rights reserved.
Objective: To revisit the mechanism of action of enfuvirtide (T20) and based on the newly defined mechanism, design an analogous peptide of T20 with improved antiviral activity.Design: We compared the inhibitory activity of T20 with that of T1144 on six-helix bundle (6HB) formation at different time after coculture of HIV type 1 (HIV-1) envelope (Env)-expressing Chinese hamster ovary (CHO-Env) cells and CD4(+)-expressing MT-2 cells at 31.5 degrees C and with that of T20-SF, an analogous peptide of T20 with an additional tryptophan-rich motif, on hemolysis mediated by FP-P, which contains fusion peptide and fusion peptide (FP) proximal region (FPPR), and HIV-1 infection.Methods: Inhibitory activity of peptides on 6HB formation was tested in a temperature-controlled cell-cell fusion assay by flow cytometry using 6HB-specific mAb 2G8; on HIV-1 infection and fusion was assessed by p24 and cell-cell fusion assays. Interaction between different peptides or peptide and antibody was evaluated by ELISA.Results: T20 could inhibit 6HB formation at early, but not late, stage of HIV-1 fusion, whereas T1144 was effective at both stages. T20-SF is much more effective than T20 in binding to FP-P and inhibiting infection of HIV-1, including T20-resistant strains, and FP-P-mediated hemolysis.Conclusion: Results suggest that T20 has a double-target mechanism, by which its N-terminal and C-terminal portions bind to N-terminal heptad repeat and FPPR, respectively. T20-SF designed based on this new mechanism exhibits significantly improved anti-HIV-1 activity because it targets the triple sites in gp41, including N-terminal heptad repeat, FPPR, and fusion peptide. Thus, this study provides clues for designing novel HIV fusion inhibitors with improved antiviral activity. Copyright (C) 2019 Wolters Kluwer Health, Inc. All rights reserved.