Multivalent Env gp120 Targeting Gold Nanoparticle Virucides of HIV-1

描述（由申请人提供）：该项目的主要目标是鉴定HIV-1拮抗剂，这些拮抗剂可以在宿主细胞接触前特异性地抑制病毒。抑制HIV-1最初进入宿主细胞仍然是预防病毒感染和传播的一种引人注目但又难以捉摸的手段。进入取决于病毒包膜蛋白刺突在触发构象的多阶段过程中与特定细胞受体相互作用的能力 Env中的重排以及随后的病毒和细胞膜的融合以将病毒内容物递送至宿主。肽三唑类进入抑制剂先前已被鉴定，并发现能够以纳摩尔亲和力结合HIV-1 gp 120，抑制Env蛋白在其CD 4和辅助受体结合位点的蛋白配体相互作用，并抑制广泛的病毒亚型的细胞感染。引人注目的是，我们最近发现，在金纳米颗粒（AuNPs）上展示的肽三唑KR 13的多价形式能够在不存在宿主细胞的情况下破坏病毒颗粒，导致内部蛋白p24的泄漏并表现出强的抗病毒活性。我们假设，多价肽三唑金纳米颗粒利用病毒Env的内在亚稳态，并且基于调节其基本特性来优化这种纳米组装体可以帮助识别HIV-1特异性杀病毒剂，以通过促进无细胞病毒破裂和灭活来用于艾滋病治疗和预防。基于这些发现，本研究的具体目标是：（1）确定纳米金-肽三唑（AuNP-PT）促进无细胞HIV-1病毒裂解的空间几何形状和表面刚性的规律;（2）确定AuNP-PT诱导HIV-1病毒裂解的机制特性及其与病毒细胞感染致病过程的关系;（3）建立AuNP-KR 13纳米结构的基本稳定性和细胞转运性质。总体而言，这项工作将得出设计多价Env靶向NP的原则，以实现对HIV-1特异性的杀病毒作用。NP将有助于扩大对Env亚稳定性的理解，该亚稳定性对于致病性宿主细胞进入至关重要，可以被劫持以开发用于HIV-1治疗剂和杀微生物剂的工具。这些结果将为如何设计其他gp 120转运蛋白-NP组合物用于HIV-1病毒灭活以及如何为含有亚稳态融合前表面蛋白复合物的其他病毒潜在地实现配体特异性病原体破裂提供先例。

DESCRIPTION (provided by applicant): This project addresses a major goal to identify HIV-1 antagonists that could inactivate the virus specifically before host cell encounter. Inhibition of he initial entry of HIV-1 into host cells remains a compelling and yet elusive means to prevent infection and spread of the virus. Entry is dependent on the ability of the virus envelope protein spike to interact with specific cell receptors in a multistage process that triggers conformational rearrangements in Env and consequent fusion of virus and cell membrane to deliver virus contents to the host. A peptide triazole class of entry inhibitors has been identified previously and found to be able to bind to HIV-1 gp120 with nanomolar affinity, to suppress protein ligand interactions of the Env protein at both its CD4 and co-receptor binding sites and to inhibit cell infection by a broad range of virus subtypes. Strikingly, we have found recently that a multivalent form of the peptide triazole KR13 displayed on gold nanoparticles (AuNPs) is able to disrupt virus particles in the absence of host cells, causing leakage of the internal protein p24 and exhibiting strong antiviral activity. We hypothesize that the multivalent peptide triazole AuNPs are capitalizing on the intrinsic metastability of the virus Env, and that optimizing such nanoassemblies based on modulating their fundamental properties can help identify HIV-1-specific virucidal agents to use in AIDS treatment and prevention by promoting cell-free virus rupture and inactivation. Based on these findings, the Specific Aims of this proposed project are to (1) determine rules of spatial geometry and surface rigidity of gold nanoparticle - peptide triazole (AuNP-PT) that promote cell-free HIV-1 virolysis; (2) determine mechanistic properties of AuNP-PT induced HIV-1 virolysis and its relationship to the pathogenic process of virus cell infection; (3) establish fundamental stability and cell transport properties of AuNP-KR13 nanoconstructs. Overall, this work will derive principles for designing multivalent Env-targeting NP's to enable virucidal actions that are specific for HIV-1. The NP's will help expand understanding of the extent to which the Env metastability that is critical for pathogenic host cel entry can be hijacked to develop tools for HIV-1 therapeutics and microbicides. The results will provide precedent for how other gp120 inhibitor-NP compositions can be devised for HIV-1 virus inactivation as well as how ligand-specific pathogen rupture can potentially be achieved for other viruses that contain metastable prefusion surface protein complexes.