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D-peptide inhibitors of HIV assembly and maturation

HIV 组装和成熟的 D 肽抑制剂

基本信息

批准号：
8020942
负责人：
WUYUAN LU
金额：
$ 18.56万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-15 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8020942
关键词：
Acquired Immunodeficiency Syndrome Affinity Amino Acids Anti-Retroviral Agents Antiviral Agents Bacteriophages Binding Biological Availability C-terminal Capsid Proteins Cells Cessation of life Chemicals Drug Compounding Drug Kinetics Drug resistance Environment Enzymes Epidemic Gagging HIV HIV Infections HIV drug resistance HIV-1 Image In Vitro Infection Integrase Lead Libraries Life Ligands Ligation Mediating Modification Patients Peptide Hydrolases Peptide Library Peptide Synthesis Peptides Peripheral Blood Mononuclear Cell Phage Display Pharmaceutical Preparations Phase Process Proteins Proteolysis RNA-Directed DNA Polymerase Regimen Research Research Design Resistance Solid Specificity Testing Therapeutic Therapeutic Agents Toxic effect Translating Viral Viral Load result Virus Assembly antiretroviral therapy base combat design drug development enantiomer fighting gag Gene Products immunogenicity improved in vitro activity inhibitor/antagonist insight novel particle peptide L peptide chemical synthesis preclinical study prevent public health relevance small molecule success therapeutic development uptake weapons

项目摘要

DESCRIPTION (provided by applicant): D-Peptide Inhibitors of HIV Assembly and Maturation Background: New therapies less susceptible to drug resistance are needed to combat the global HIV epidemic. Virus assembly and maturation remains a significant yet largely unexploited antiviral target. HIV assembly and maturation is mediated at the protein level by self-association of the HIV Gag polyprotein and of its sub-domains such as the matrix protein (MA) and capsid protein (CA). Peptides derived from MA and CA or selected from phage libraries have been shown to be able to block virus assembly and maturation in vitro. However, the antiviral effects of these peptide-based assembly/maturation inhibitors vary, due to a large extent to their poor stability in the protease-rich environment of the living cell - an inherent drawback of peptide therapeutics. Peptides composed entirely of D-amino acids, i.e., D-peptides, are resistant to proteolysis, which translates into much-improved bioavailability and reduced immunogenicity as compared with conventional L- peptides. Thus, D-peptides are ideally suited as lead drug compounds for therapeutic development. Objective/Hypothesis: We seek to develop, by the means of chemical protein synthesis and mirror-image phage display, D-peptide based antagonists of HIV MA and the C-terminal domain of CA (C-CA) as a novel class of therapeutic agents for the treatment of HIV-1 infection. Specific Aims: (1) Synthesize the D- enantiomers of HIV-1 MA and C-CA composed entirely of D-amino acids using a combination of solid phase peptide synthesis and native chemical ligation; (2) Screen phage-expressed peptide libraries against DMA and DC-CA and identify optimal sequences required for productive binding to the D-proteins; (3) Test the hypothesis that D-peptide antagonists of native MA and C-CA inhibit HIV replication in peripheral blood mononuclear cells. Study design: We will screen phage-expressed peptide libraries against chemically synthesized DMA and DC-CA, identifying high-affinity binders for the D-proteins. Inversion from L to D through peptide synthesis will convert phage-optimized L-peptide ligands to their mirror image D-forms that specifically bind native LMA and LC-CA. We will structurally characterize the interaction between the D-peptides and LMA or LC-CA, and evaluate their antiviral activity in vitro. In addition, mechanistic studies will be carried out to examine how the D-peptides disrupt HIV Gag assembly. Finally, novel delivery vehicles will be designed to improve peptide uptake by target cells. Significance: The proposed research seeks to fill the obvious gap by developing D-peptide inhibitors to prevent assembly of both immature and mature HIV particles. The specific aims outlined in this proposal, if achieved, will enable us to design D-peptide antiviral therapeutics for preclinical studies. This novel class of peptide antagonists, emulating the activity of known assembly/maturation inhibitors, enjoys high specificity and less toxicity compared with traditional small- molecule drugs, and is superior to conventional L-peptide-based antiviral agents with respect to bioavailability, pharmacokinetics, and immunogenicity. Identification of D-peptide antagonists may help better understand the virus assembly and maturation process and illuminate insight as well into designing other classes of antagonistic molecules to inhibit HIV replication. PUBLIC HEALTH RELEVANCE: Current antiretroviral therapy (ART) for HIV-1 infected patients utilizes a combination of inhibitors that target the viral enzymes reverse transcriptase and protease. ART reduces viral load and slows the progression of HIV to AIDS, contributing to a steady decrease in AIDS deaths in the world. Despite its success, ART does not eradicate HIV from infected cells, and, among many complications of ART is the emergence of drug-resistant HIV strains not responding to current antiretroviral regimens. The proposed research aims to develop a novel class of D-peptide inhibitors as additional weapons in the arsenal to fight HIV infection by specifically targeting HIV assembly and maturation.

描述（由申请人提供）：HIV 组装和成熟的 D 肽抑制剂背景：需要不易产生耐药性的新疗法来对抗全球 HIV 流行。病毒组装和成熟仍然是一个重要但很大程度上尚未开发的抗病毒靶点。 HIV 的组装和成熟是在蛋白质水平上通过 HIV Gag 多蛋白及其子结构域（例如基质蛋白 (MA) 和衣壳蛋白 (CA)）的自缔合介导的。源自 MA 和 CA 或选自噬菌体文库的肽已被证明能够在体外阻断病毒组装和成熟。然而，这些基于肽的组装/成熟抑制剂的抗病毒作用各不相同，很大程度上是由于它们在活细胞富含蛋白酶的环境中稳定性差，这是肽治疗的固有缺陷。完全由D-氨基酸组成的肽，即D-肽，具有抗蛋白水解作用，与传统的L-肽相比，这意味着生物利用度大大提高，免疫原性降低。因此，D-肽非常适合作为治疗开发的先导药物化合物。目的/假设：我们寻求通过化学蛋白质合成和镜像噬菌体展示的方式开发基于 D 肽的 HIV MA 和 CA 的 C 末端结构域 (C-CA) 拮抗剂，作为治疗 HIV-1 感染的新型治疗药物。具体目标：（1）采用固相肽合成和天然化学连接相结合的方式合成完全由D-氨基酸组成的HIV-1 MA和C-CA的D-对映体； (2) 针对 DMA 和 DC-CA 筛选噬菌体表达的肽文库，并确定与 D 蛋白有效结合所需的最佳序列； (3)检验天然MA和C-CA的D肽拮抗剂抑制外周血单核细胞中HIV复制的假设。研究设计：我们将针对化学合成的 DMA 和 DC-CA 筛选噬菌体表达的肽库，鉴定 D 蛋白的高亲和力结合物。通过肽合成从 L 到 D 的反转会将噬菌体优化的 L 肽配体转化为其镜像 D 形式，从而特异性结合天然 LMA 和 LC-CA。我们将从结构上表征 D 肽与 LMA 或 LC-CA 之间的相互作用，并评估它们的体外抗病毒活性。此外，还将进行机制研究以检验 D 肽如何破坏 HIV Gag 组装。最后，将设计新型递送载体以改善靶细胞对肽的摄取。意义：拟议的研究旨在通过开发 D 肽抑制剂来防止未成熟和成熟 HIV 颗粒的组装来填补明显的空白。该提案中概述的具体目标如果实现，将使我们能够设计用于临床前研究的 D 肽抗病毒疗法。这类新型肽拮抗剂模仿已知组装/成熟抑制剂的活性，与传统小分子药物相比具有较高的特异性和较低的毒性，并且在生物利用度、药代动力学和免疫原性方面优于传统的基于L-肽的抗病毒药物。 D 肽拮抗剂的鉴定可能有助于更好地了解病毒组装和成熟过程，并阐明设计其他类别的拮抗分子来抑制 HIV 复制的见解。公共卫生相关性：目前针对 HIV-1 感染患者的抗逆转录病毒疗法 (ART) 采用针对病毒酶逆转录酶和蛋白酶的抑制剂组合。抗逆转录病毒疗法可减少病毒载量并减缓艾滋病毒向艾滋病的进展，从而有助于世界各地艾滋病死亡人数的稳步下降。尽管 ART 取得了成功，但它并没有从受感染的细胞中根除 HIV，而且 ART 的许多并发症之一是出现了对当前抗逆转录病毒疗法没有反应的耐药 HIV 毒株。拟议的研究旨在开发一类新型 D 肽抑制剂，作为武器库中的额外武器，通过专门针对 HIV 组装和成熟来对抗 HIV 感染。