D-peptide inhibitors of HIV assembly and maturation

HIV 组装和成熟的 D 肽抑制剂

• 批准号: 7929943
• 负责人: WUYUAN LU
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-15 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929943
• 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; Protein C; 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; matrix protein, Human immunodeficiency virus type 1; novel; particle; peptide L; peptide chemical synthesis; preclinical study; prevent; public health relevance; small molecule; success; therapeutic development; uptake; weapons; 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; Protein C; 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; matrix protein, Human immunodeficiency virus type 1; novel; particle; peptide L; peptide chemical synthesis; preclinical study; prevent; public health relevance; small molecule; success; therapeutic development; uptake; weapons

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组装和成熟背景的D肽抑制剂：与全球HIV流行作斗争所需的新疗法不易受到耐药性。病毒组装和成熟仍然是一个重要但没有开发的抗病毒靶标。 HIV组装和成熟是通过HIV GAG多蛋白及其亚构域（例如基质蛋白（MA）和衣壳蛋白（CA）的亚植物的自我关联（CA）的自我关联，在蛋白质水平上介导。源自MA和CA或从噬菌体文库中选择的肽已被证明能够在体外阻断病毒组装和成熟。但是，这些基于肽的组装/成熟抑制剂的抗病毒作用因其在活细胞的蛋白酶丰富的环境中的稳定性很大而变化，这是肽疗法的固有缺点。与常规的L-肽相比，完全由D-氨基酸组成的肽，即D肽，对蛋白水解具有抗药性，它转化为备受改善的生物利用度和降低的免疫原性。因此，理想地将D肽作为治疗性发育的铅药物化合物。客观/假设：我们试图通过化学蛋白合成和镜像噬菌体显示，基于D肽的HIV MA的拮抗剂和Ca（C-CA）的C末端结构域（C-CA）作为新型治疗剂治疗HIV-1感染的新型治疗剂。具体目的：（1）合成HIV-1 MA和C-CA的D-剂，完全由D-氨基酸组成，结合了固相肽合成和天然化学连接； （2）针对DMA和DC-CA的屏幕噬菌体表达的肽库，并确定生产性结合与D蛋白所需的最佳序列； （3）检验以下假设：天然MA和C-CA的D肽拮抗剂抑制外周血单核细胞中的HIV复制。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，从而鉴定D蛋白的高亲和力粘合剂。通过肽合成从L到D的反转将将噬菌体优化的L肽配体转换为特异性结合天然LMA和LC-CA的镜像D形式。我们将在结构上表征D肽与LMA或LC-CA之间的相互作用，并在体外评估其抗病毒活性。此外，将进行机械研究，以检查D肽如何破坏HIV GAG组装。最后，新型的输送车将旨在改善目标细胞的肽摄取。意义：拟议的研究旨在通过开发D肽抑制剂来防止同时组装不成熟和成熟的HIV颗粒，以填补明显的空白。如果达成的话，该提案中概述的具体目的将使我们能够为临床前研究设计D肽抗病毒疗法。与传统的小分子药物相比，这种新型的肽拮抗剂模拟已知的组装/成熟抑制剂的活性具有很高的特异性和较小的毒性，并且在传统的基于L肽的抗病毒剂方面相对于生物通用性，药物动力学和免疫基因而言优越。 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，并阐明洞察力以及设计其他类别的拮抗分子以抑制HIV复制。 公共卫生相关性：HIV-1感染患者的当前抗逆转录病毒疗法（ART）利用靶向病毒酶逆转录酶和蛋白酶的抑制剂组合。 ART减少了病毒载荷并减慢了艾滋病毒向艾滋病的发展，从而导致世界上艾滋病死亡的稳定减少。尽管取得了成功，但ART并未从感染细胞中消除HIV，并且在许多艺术并发症中，抗药性HIV菌株的出现不反应当前的抗逆转录病毒治疗方案。拟议的研究旨在开发一种新型的D肽抑制剂，作为阿森纳的其他武器，通过专门针对HIV组装和成熟来抵抗HIV感染。