Development of Therapeutics to Eliminate HIV

描述(申请人提供)：该项目的长期目标是开发将导致消除人类免疫缺陷病毒的治疗方法。这一特殊应用的目标是开发特洛伊木马抑制剂(THIS)，它是实现我们长期目标的第一步，它由HIV蛋白酶激活，用于杀死HIV试图在其中复制的细胞。中心假设是，消除激活病毒的细胞可以防止病毒复制。第一代THI是一个由四部分组成的蛋白质：细胞穿透序列、膜破坏肽、阻断膜破坏肽作用的掩蔽螺旋以及位于掩蔽螺旋和膜破坏肽之间的HIV蛋白酶裂解位点。细胞穿透序列促进了细胞的吸收。在没有艾滋病毒的情况下，什么都不会发生。在HIV存在的情况下，该肽被HIV蛋白酶裂解，释放出膜破坏肽，该肽破坏膜功能，导致细胞死亡，不产生病毒。将检验中心假设，并通过追求以下特定目标来实现这一应用的总体目标：1.确定其体外功能。第一代和后续代的功能的几个重要方面这可以在体外测试，包括正确的HIV蛋白酶切割，HIV蛋白酶激活和膜破坏2.研究第一代THI在组织培养中杀灭HIV感染细胞和减少病毒产生的能力。THI应该在组织培养中发挥作用，杀死艾滋病毒感染的细胞，并保留未感染的细胞。3.设计THI的第二代和后续世代。THI由四个元件组成：一个多肽摄取序列、一个膜破坏多肽、一个掩蔽螺旋和HIV蛋白酶位点。利用基于知识的设计以及体外和组织培养测试的结果，将对这些细胞进行合理的功能优化。该项目的主要创新是将常规特洛伊木马策略(引入由病毒酶激活的良性物质)与依赖于膜破坏的生物物理杀灭机制相结合。特洛伊木马抑制剂有可能阻止病毒的产生，因此可能提供一条治愈艾滋病的途径。自1981年被确认为流行病以来，艾滋病已导致2500万人死亡。 与公共卫生相关：目前有近4300万人感染艾滋病毒/艾滋病。2005年有310万人死于艾滋病，460万人新感染。艾滋病是世界范围内的一个主要卫生保健问题。这项拟议的研究意义重大，因为它涉及到可能导致艾滋病治愈方法开发的治疗方法的发展。

DESCRIPTION (provided by applicant): The long term goal of this project is to develop therapeutics that will result in the elimination of human immunodeficiency virus. The objective of this particular application, which represents a first step toward the attainment of our long term goal, is the development of Trojan horse inhibitors (THIs) that are activated by HIV protease and serve to kill the cells in which the HIV is attempting to replicate. The central hypothesis is that elimination of the cells in which the virus is activated can prevent viral replication. The first generation THI is a protein consisting of four parts: a cellular penetration sequence, a membrane-disruption peptide, a masking helix that blocks the action of the membrane-disruption peptide and a HIV protease cleavage site between the masking helix and the membrane- disruption peptide. The cellular penetration sequence promotes cellular uptake. In the absence of HIV, nothing happens. In the presence of HIV, the peptide is cleaved by HIV protease releasing the membrane-disruption peptide, the peptide disrupts membrane function leading to cell death and virus is not produced. The central hypothesis will be tested and the overall objectives of this application accomplished by pursuing the following specific aims: 1. Determine the in vitro functionality of THIs. Several important aspects of the functionality of the first and subsequent generation THIs can be tested in vitro including correct HIV protease cleavage, HIV protease activation and membrane disruption 2. Investigate the ability of the first generation THI to kill HIV infected cells and reduce virus production in tissue culture. THI should function in tissue culture to kill HIV infected cells and spare non-infected cells. 3. Design the second and subsequent generations of THI. The THI consists of four elements: a peptide uptake sequence, a membrane-disruptive peptide, a masking helix and the HIV protease site. These will be rationally optimized for function using knowledge-based design and the results of in vitro and tissue culture testing. The main innovation of this project is to combine the use of a general Trojan horse strategy (introducing a benign substance that is activated by a virus enzyme) with a biophysical killing mechanism that relies on membrane disruption. Trojan horse inhibitors have the potential to stop viral production and so could provide a route to a cure for AIDS. AIDS has killed 25 million people since being recognized as an epidemic in 1981. PUBLIC HEALTH RELEVANCE: There are almost 43 million people currently living with HIV/AIDS. There were 3.1 million deaths from AIDS and 4.6 million new infections in 2005. AIDS is a major health care problem worldwide. The proposed research is significant because it concerns the development of therapeutics that could lead to the development of a cure for AIDS.