Multivalent Env gp120 Targeting Gold Nanoparticle Virucides of HIV-1

描述(由申请人提供)：该项目的一个主要目标是确定HIV-1拮抗剂，这些拮抗剂可以在宿主细胞相遇之前特定地灭活病毒。抑制艾滋病毒-1最初进入宿主细胞仍然是防止病毒感染和传播的一种令人信服但又难以捉摸的手段。进入依赖于病毒包膜蛋白尖峰与特定细胞受体在触发构象的多阶段过程中相互作用的能力 在环境中的重排和随后的病毒和细胞膜的融合，将病毒内容传递到宿主。三唑类多肽进入抑制剂已被发现能够以纳摩尔亲和力与HIV-1 gp120结合，在其CD4和辅助受体结合部位抑制Env蛋白的蛋白质配体相互作用，并抑制广泛的病毒亚型对细胞的感染。值得注意的是，我们最近发现，在金纳米颗粒(AuNPs)上展示的多价形式的三氮唑KR13能够在没有宿主细胞的情况下破坏病毒颗粒，导致内部蛋白p24的泄漏，并显示出强大的抗病毒活性。我们假设多价多肽三氮唑AuNPs利用了病毒环境的内在亚稳性，并且基于调节它们的基本性质来优化这种纳米组件可以通过促进无细胞病毒破裂和灭活来帮助识别用于艾滋病治疗和预防的HIV-1特异性杀病剂。基于这些发现，这项拟议项目的具体目标是(1)确定促进无细胞HIV-1病毒裂解的金纳米颗粒-多肽三唑(AuNP-PT)的空间几何和表面刚性规则；(2)确定AuNP-PT诱导的HIV-1病毒裂解的机制性质及其与病毒细胞感染的致病过程的关系；(3)建立AuNP-KR13纳米结构的基本稳定性和细胞运输性质。总体而言，这项工作将得出设计多价环境靶向NP的原则，以实现针对HIV-1的杀毒作用。NP‘s将有助于扩大对环境亚稳性的理解，环境亚稳性对致病宿主细胞进入至关重要，可以被劫持以开发HIV-1治疗和杀微生物剂的工具。这些结果将为如何设计其他gp120抑制剂-NP组合物用于HIV-1病毒灭活以及如何潜在地为包含亚稳态预融合表面蛋白复合体的其他病毒实现配体特异性病原体断裂提供先例。

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.