Immunology, cell biology, and mechanism of biguanide

免疫学、细胞生物学和双胍的作用机制

• 批准号: 6809134
• 负责人: Fred C. Krebs
• 金额: $ 8.68万
• 依托单位: NOVAFLUX BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6809134
• 关键词: antiAIDS agent; biological signal transduction; cell migration; cervix; chemokine receptor; cooperative study; cytokine; drug design /synthesis /production; drug screening /evaluation; gene expression; guanidines; human immunodeficiency virus 1; human tissue; inflammation; laboratory mouse; laser capture microdissection; microarray technology; pharmacokinetics; topical drug application; vagina; virus infection mechanism

The goal of studies in Project II is to define the safety (in vitro cytotoxicity, in vivo toxicity, and immunogenicity) and mechanisms of action that underlie the anti-HIV-1 activities of compounds collectively classified as polybiguanides (PBGs). The hypotheses that form the basis of Project II are that (i) PEHMB-based compounds (or combinations containing PEHMB) can be designed, synthesized, and formulated with little or no toxicity or immunogenicity, and (ii) the mechanism of action of PEHMB against HIV-1 infection involves direct, specific interactions with HIV-1 co-receptors CXCR4 and CCR5. Our studies have identified polyethylene hexamethylene biguanide (PEHMB) as a prototypical PBG with low cytotoxicity, substantial anti-HIV-1 activity, and an in vitro therapeutic index in excess of 1400. The PEHMB molecule will be used as a candidate microbicide as well as the cornerstone for logical, computer-aided design and synthesis strategies directed toward optimizing the efficacy of PEHMB derivatives. Our efforts, which will culminate in demonstrations of in vivo efficacy against HIV-1 in animal models of HIV-1 infection (Project III), will serve as a foundation for clinical trials of highly efficacious anti-HIV-1 PBG-based compounds as safe and effective topical vaginal microbicides. The specific aims of Project II are to (i) assess the effect of PEHMB and derivative compounds, combinations, and formulations (identified in Projects I and III) on the in vitro viability of cells of epithelial and immune origin found in the human female reproductive tract, and on the viability and integrity of vaginal and cervical epithelial tissues using mouse and rabbit models of in vivo toxicity; (ii) demonstrate the impact of PEHMB and select derivative compounds on gene expression patterns and cytokine/chemokine signaling in single cell populations and on immune cell recruitment and inflammatory processes within vaginal and cervical epithelial tissues in the mouse model of in viv0 toxicity (as assessed by microarray analyses and laser capture microdissection); and (iii) determine the effects of interactions between PEHMB and HIV-1 co-receptors CXCR4 and CCR5 on HIV-1 binding and entry events that precede HIV-1 infection and on co-receptor functions. This project is an integral part of highly collaborative studies focused on the development of safe and effective PBG-based microbicides with activity against HIV-I. These studies will serve to significantly advance our development of an affordable topical PBG-based microbicide used for the global reduction or elimination of HIV-1 sexual transmission.

项目II中研究的目标是确定安全性（体外细胞毒性、体内毒性和 免疫原性）和作为统称为聚双胍（PBG）的化合物的抗HIV-1活性的基础的作用机制。构成项目II基础的假设是：（i）可以设计、合成和配制具有很少或没有毒性或免疫原性的基于PEHMB的化合物（或含有PEHMB的组合），以及（ii）PEHMB抗HIV-1感染的作用机制涉及与HIV-1共受体CXCR 4和CCR 5的直接、特异性相互作用。我们的研究已经确定聚乙烯双胍（PEHMB）作为一个原型PBG具有低细胞毒性，大量的抗HIV-1活性，在体外治疗指数超过1400。PEHMB分子将用作 候选杀微生物剂以及逻辑，计算机辅助设计和合成策略的基石，旨在优化PEHMB衍生物的功效。我们的努力将最终在HIV-1感染的动物模型中证明体内抗HIV-1的有效性（项目III），这将为高效抗HIV-1 PBG化合物作为安全有效的局部阴道杀微生物剂的临床试验奠定基础。项目II的具体目标是（i）评估PEHMB和衍生化合物、组合和制剂的效果（项目I和III中确定的）对人类女性生殖道中发现的上皮细胞和免疫源细胞的体外活力，以及使用小鼠和家兔体内毒性模型对阴道和宫颈上皮组织的活力和完整性的影响;（ii）证明PEHMB和选择的衍生化合物对基因表达模式的影响， 体内毒性小鼠模型中单细胞群中的细胞因子/趋化因子信号传导以及阴道和宫颈上皮组织内的免疫细胞募集和炎症过程（如通过微阵列分析和激光捕获显微切割评估的）;和（iii）确定PEHMB与HIV-1共受体CXCR 4和CCR 5之间的相互作用对HIV-1结合和HIV-1进入事件的影响，1感染和辅助受体功能。该项目是高度合作研究的一个组成部分，重点是开发安全有效的基于PBG的杀微生物剂，对HIV-I具有活性。这些研究将大大促进我们开发一种负担得起的基于PBG的局部杀微生物剂 用于全球减少或消除HIV-1性传播。