Anti-HIV-1 activity of single & multicomponent biguanide

我们正在开发一类新型的非表面活性剂杀微生物剂，聚双胍（PBG），扰乱膜结构，重新分配膜相关蛋白（包括HIV-1共受体），并改变病毒/宿主细胞相互作用。在项目III中，我们建议评估在项目I中合成、纯化和表征的基于聚乙烯、双胍（PEHMB）的杀微生物剂和组合杀微生物剂的抗HIV-1活性。在项目三中进行测试的铅化合物将是那些先前在项目二中被评估为无毒的化合物。假设计算化学方法与合理药物设计相结合，将导致基于PEHMB的杀微生物剂的单一或组合制剂表现出低毒性和高度的抗HIV-1活性。为此，我们将PEHMB化合物与无毒、高活性的硫酸化树枝状聚合物结合，以生产通过多种作用机制阻断HIV-1感染的杀微生物剂制剂。项目III的具体目的是（1）定量未配制和配制的基于PEHMB和基于PEHMB-树枝状聚合物组合的化合物（在项目I中合成）在无细胞病毒（CFI）灭活、淋巴细胞或单核细胞来源的感染细胞内细胞相关病毒（CAI）灭活以及病毒结合和进入（VBI）抑制方面的抗HIV-1活性; 2.）确定基于PEHMB的化合物和基于PEHMB-树枝状聚合物组合的化合物（在项目I中合成）关于抑制利用无细胞和细胞相关病毒接种物体外培养的人阴道细胞靶标（淋巴细胞、单核细胞、树突细胞和上皮细胞）的HIV-1感染的功效; 3.）利用阴道上皮的体外器官培养模型来检查感染性HIV-1通过含有选择的基于PEHMB的化合物和基于PEHMB-树枝状聚合物组合的化合物（在目的1和2中鉴定）的人工阴道制剂渗透到下面的容许靶细胞单层和4。测量选择的基于PEHMB和基于PEHMB-树枝状聚合物组合的化合物（在目的1和2中鉴定）在两种病毒感染模型中阻断无细胞和细胞相关HIV-1感染的能力：小鼠阴道hu-PBL-NOD-SCID模型和NOD-SCID小鼠中的人阴道上皮组织异种移植模型。项目III中进行的研究将与项目I和项目II中的研究人员协商进行，他们将参与利用体外和体内模型系统对基于PEHMB的制剂的细胞敏感性的详细评估。项目II和III中获得的结果将用于推导项目I中构建的所选PEHMB基化合物的体外治疗指数。这些结果将可用于选择最有希望的化合物以在HIV-1感染的人阴道异种移植模型中进行检查，并用于人类临床试验的进展。

We are developing a novel class of non-surfactant microbicidal agents, the polybiguanides (PBG) that perturb membrane structure, redistribute membrane-associated proteins (including HIV-1 co-receptors and alter virus/host cell interactions. In Project III, we propose to assess the anti-HIV-1 activity of polyethylene, hexamethylene biguanide (PEHMB)-based microbicides and combination microbicides synthesized, purified, and characterized in Project I. Lead compounds for testing in Project III will be those previously assessed as non-toxic in Project II. The HYPOTHESIS is that computational chemistry approaches combined with rationale drug design will result in single or combination formulations of PEHMB-based microbicides that exhibit low toxicity with a high degree of anti-HIV-1 activity. To this end, we are combining PEHMB-compounds with non-toxic, highly active sulfated dendrimers in order to produce microbicide formulations that interdict HIV-1 infection by multiple mechanisms of action. The SPECIFIC AIMS of Project III are to (1) quantitate the anti-HIV-1 activity of non-formulated and formulated PEHMB-based and PEHMB-dendrimer combination based compounds (synthesized in Project I) with respect to inactivation of cell-free virus (CFI), inactivation of cell-associated virus (CAI) within infected cells of lymphocytic or monocytic origin and inhibition of viral binding and entry (VBI); 2.) determine the efficacy of PEHMB-based and PEHMB-dendrimer combination based compounds (synthesized in Project I) with respect to inhibiting HIV-1 infection of human vaginal cellular targets (lymphocytes, monocytes, dendritic cells, and epithelial cells) cultured in vitro utilizing both cell-free and cell-associated viral inoculums; 3.) Utilize an in vitro organ culture model of vaginal epithelium to examine the penetration of infectious HIV-1 through artificial vaginal formulations containing select PEHMB-based and PEHMB-dendrimer combination based compounds (identified in Aims 1 and 2) to an underlying permissive target cell monolayer and 4.) measure the ability of select PEHMB-based and PEHMB-dendrimer combination based compounds (identified in Aims 1 and 2) to block infections by cell-free and cell-associated HIV-1 in two models of virus infection: a mouse vaginal hu-PBL-NOD-SCID model and a human vaginal epithelial tissue xenograft model in NOD-SCID mice. The studies _erformed in Project III will be pursued in consultation with investigators in Project I and Project II, who will be involved in a detailed assessment of the cellular sensitivity of PEHMB-based formulations utilizing both in vitro and in vivo model systems. Results obtained in Projects II and III will be utilized to derive in vitro therapeutic indices for the selected PEHMB-based compounds that will be constructed in Project I. These results will be useful in selecting the most promising compounds to examine in the HIV-l-infected human vaginal xenograft model and for advancement to clinical trials in humans.