Resistance Profiles of Novel Electrostatically Constrained HIV-1 Fusion Inhibitors*
Resistance Profiles of Novel Electrostatically Constrained HIV-1 Fusion Inhibitors*
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DOI：
10.1074/jbc.m110.145789
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发表时间：
2010-10
期刊：
影响因子：
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通讯作者：
Kazuya Shimura;Daisuke Nameki;Keiko Kajiwara;Kentaro Watanabe;Yasuko Sakagami;S. Oishi;N. Fujii;
中科院分区：
文献类型：
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作者：
Kazuya Shimura;Daisuke Nameki;Keiko Kajiwara;Kentaro Watanabe;Yasuko Sakagami;S. Oishi;N. Fujii;
人类免疫缺陷病毒（HIV）gp 41在病毒融合中起关键作用; gp 41的N-和C-末端七肽重复序列（N-HR和C-HR）形成稳定的6-螺旋构象用于融合。因此，HR衍生的肽，如恩夫韦肽（T-20），通过充当诱饵来抑制HIV-1融合，并已用于治疗HIV-1感染。然而，T-20的疗效因gp 41中的耐药突变而减弱，包括V38 A和N43 D。为了抑制耐药变异体，我们先前开发了静电约束肽SC 34和SC 34 EK，并显示两者对野生型和T-20耐药变异体表现出有效的抗HIV-1活性。在这项研究中，为了阐明对下一代融合抑制剂的耐药机制，我们在体外选择了对SC 34和SC 34 EK具有耐药性的变体。耐药变体在gp 41中具有多个突变。所有这些突变单独引起对SC 34和SC 34 EK的小于6倍的抗性，表明高水平抗性存在显著的遗传障碍。通过两个分子内静电对的极性的简单差异降低了对SC 34和SC 34 EK的交叉抗性。此外，所选择的突变增强了与N-HR变体的物理化学相互作用，并恢复了亲本肽C34的活性，甚至对抗性变体也是如此。这些结果表明，我们的方法设计的gp 41结合抑制剂，利用静电约束和来自耐药性研究的信息产生抑制剂与T-20和其他抑制剂的活性增强，高遗传屏障，和不同的耐药性。因此，这是一个有前途的方法，为未来一代肽融合抑制剂的设计。
Human immunodeficiency virus (HIV) gp41 plays a key role in viral fusion; the N- and C-terminal heptad repeats (N-HR and C-HR) of gp41 form a stable 6-helical conformation for fusion. Therefore, HR-derived peptides, such as enfuvirtide (T-20), inhibit HIV-1 fusion by acting as decoys, and have been used for the treatment of HIV-1 infection. However, the efficacy of T-20 is attenuated by resistance mutations in gp41, including V38A and N43D. To suppress the resistant variants, we previously developed electrostatically constrained peptides, SC34 and SC34EK, and showed that both exhibited potent anti-HIV-1 activity against wild-type and T-20-resistant variants. In this study, to clarify the resistance mechanism to this next generation of fusion inhibitors, we selected variants with resistance to SC34 and SC34EK in vitro. The resistant variants had multiple mutations in gp41. All of these mutations individually caused less than 6-fold resistance to SC34 and SC34EK, indicating that there is a significant genetic barrier for high-level resistance. Cross-resistance to SC34 and SC34EK was reduced by a simple difference in the polarity of two intramolecular electrostatic pairs. Furthermore, the selected mutations enhanced the physicochemical interactions with N-HR variants and restored activities of the parental peptide, C34, even to resistant variants. These results demonstrate that our approach of designing gp41-binding inhibitors using electrostatic constraints and information derived from resistance studies produces inhibitors with enhanced activity, high genetic barrier, and distinct resistance profile from T-20 and other inhibitors. Hence, this is a promising approach for the design of future generation peptide fusion inhibitors.