Genome Targeted Inhibitors of Retroviruses

描述（由申请人提供）：现在比以往任何时候都更加强调需要针对病毒生命周期的几个阶段并且可能不易受病毒遗传灵活性影响的新型疗法，主要是因为HIV- 1耐药株的快速出现。选择的靶标开始缩小到病毒基因组的非翻译5'区，其含有多个调节元件，如TAR、PBS、A环、DIS等，这些元件对于病毒复制至关重要并且不愿突变变化。选择性干预这些调控区域的功能可能在阻断病毒感染方面具有深远的治疗潜力。我们已经确定了一些领先的聚酰胺核苷酸类似物（PNA），可以成功地阻止这些目标在体外的功能。该建议的主要目的是将这些主要的PNA与许多膜转运肽缀合，以鉴定最有效的生物递送系统，从而增强这些化合物的功能功效。将深入检查每种PNA-肽缀合物在细胞培养物中的摄取动力学、功能功效、细胞毒性和抗病毒活性。还将在动物模型中测试主要PNA-肽缀合物的药代动力学行为、组织分布和毒理学性质。随后将在用人PBL重建的hu-SCID小鼠模型上对这些化合物进行临床前试验。这些研究将提供关于这类化合物的宝贵信息，这可能有助于开发高治疗指数的有效多叉抑制剂。提出了以下具体目标。目标1：靶向HIV-1 RNA基因组关键区域的PNA-膜转运蛋白肽缀合物的设计和合成目的2：PNA-转运蛋白肽缀合物的生物递送和功能表征的评价。目的3：在细胞培养中评价PNA-肽缀合物的抗病毒效力和细胞毒性。目的4：主要PNA转运蛋白肽制剂的药代动力学分析、组织分布和毒理学评价。目的5：使用SCID-hu小鼠模型评价PNA-肽缀合物的抗病毒效力。

DESCRIPTION (provided by applicant): The need for novel therapies that target several stages of the viral life cycle and that may not be vulnerable to the genetic flexibility of the virus are now emphasized more than ever, primarily because of the rapid emergence of drug resistant strains of HIV- 1. The target of choice is beginning to be narrowed to the non-translated 5' region of the viral genome that contains multiple regulatory elements such as TAR, PBS, A-loop, DIS etc which are critical for viral replication and averse to mutational changes. Selective intervention of the function of these regulatory regions may have profound therapeutic potential in blocking the viral infection. We have identified a number of leading polyamide nucleotide analogs (PNA) which can successfully block the function of these targets in vitro. The major thrust of this proposal is to conjugate these leading PNAs with a number of membrane transporting peptides in order to identify the most efficient biodelivery system thus enhancing the functional efficacy of these compounds. Each PNA-peptide conjugate will be examined in-depth with respect to its uptake kinetics, functional efficacy, cytotoxicity and antiviral activity in cell cultures. The leading PNA-peptideconjugates will also be tested for their pharmacokinetic behaviour, tissue distribution and toxicological properties in animal models. This will be followed by subjecting these compounds to pre-clinical trial on hu-SCID mice model reconstructed with human PBL. These studies will provide invaluable information on this class of compounds which may help in the development of effective multiprong inhibitors of high therapeutic index. The following specific aims are proposed. Aim 1: Design and synthesis of PNA-membrane transporter peptide conjugates targeted to critical regions of HIV-1 RNA genome Aim 2: Evaluation of biodelivery and functional characterization of PNA- transporter peptide conjugates. Aim 3: Evaluation of antiviral efficacy and cytotoxicity of PNA-peptide conjugates in cell culture. Aim 4: Pharmacokinetic analysis, tissue distribution and toxicological evaluation of leading PNA-transporter peptide formulations. Aim 5: Evaluation of antiviral efficacy of PNA-peptide conjugates using SCID-hu mice model.