Structure Based Themodynamic Studies of HIV-1 Protease

描述（由申请人提供）：蛋白酶抑制剂是HIV-1感染化疗的基本成分。尽管抗逆转录病毒疗法取得了成功，但由于出现了对蛋白酶抑制剂具有耐药性的病毒株，其长期疗效不断受到阻碍。在接受高效抗逆转录病毒治疗（HAART）的患者中，与严重副作用发生相关的治疗失误往往会加速耐药性的发生。此外，在非洲流行的病毒亚型，绝大多数艾滋病毒感染发生在非洲，与导致美洲和欧洲感染的病毒亚型不同。使事情更加复杂的是，另一种艾滋病毒，HIV-2，虽然不如HIV-1流行，但也能引起艾滋病。很明显，成功的蛋白酶抑制剂必须针对广泛的靶标变异性保持适当的效力。截至目前，FDA已批准9种蛋白酶抑制剂用于临床。虽然它们都靶向相同酶的活性位点口袋，但它们具有不同的效力，不同的抗性谱和对不需要的靶标的不同选择性。理想的抑制剂应该对野生型蛋白酶具有极高的效力，表现出对与耐药性相关的蛋白酶突变的低敏感性，并且不干扰人类靶标，从而使副作用最小化。该项目的主要目标是开发精确的热力学和结构指南，以开发此类抑制剂。该项目的具体目标是：-发展热力学和结构规则，旨在实现极高的亲和力。- 鉴定赋予蛋白酶抑制剂对突变的低敏感性和对不同病毒亚型（包括HIV-2）的有效性的热力学和结构决定因素。- 发展热力学和结构规则，旨在限制蛋白酶抑制剂对不需要的靶点的亲和力，从而提高选择性。这些目标将通过实验热力学测量（高灵敏度等温滴定量热法和高灵敏度差示扫描量热法）、结构测定（X射线晶体学）和基于结构的热力学分析的组合来实现。 公共卫生相关性：全世界有3 000多万人感染艾滋病毒/艾滋病，每年有200多万人死亡。尽管抗逆转录病毒疗法取得了初步成功，但由于出现耐药性病毒株和出现严重副作用，这种疗法受到阻碍。该项目的主要目标是为开发抗逆转录病毒药物，特别是蛋白酶抑制剂制定精确的指导方针，其特点是效力极高，耐药性敏感性低，副作用最小。

DESCRIPTION (provided by applicant): Protease inhibitors are essential components in the chemotherapy of HIV-1 infection. Despite their success, the long-term efficacy of antiretroviral therapies is continuously hindered by the emergence of viral strains that exhibit resistance to protease inhibitors. The onset of drug resistance is often accelerated by therapy lapses associated with the occurrence of severe side effects in patients undergoing highly active anti-retroviral therapy (HAART). In addition, the viral subtypes prevalent in Africa, where the vast majority of HIV infections take place, are not the same as the one responsible for the infections in America and Europe. Complicating things even further, a different HIV virus, HIV-2, although less prevalent than HIV-1, is also able to cause AIDS. It is clear that successful protease inhibitors would have to maintain appropriate potency against a wide range of target variability. As of today, the FDA has approved nine protease inhibitors for clinical use. While all of them target the active site pocket of the same enzyme, they do so with different potency, different resistance profiles and different selectivity towards unwanted targets. An ideal inhibitor should have extremely high potency against the wild type protease, exhibit low susceptibility to protease mutations associated with drug resistance and not interfere with human targets, thus minimizing side effects. The main goal of this project is to develop precise thermodynamic and structural guidelines to develop such inhibitors. The specific goals of this project are: - Development of thermodynamic and structural rules aimed at achieving extremely high affinity. - Identification of thermodynamic and structural determinants that confer protease inhibitors low susceptibility to mutations and efficacy against different viral subtypes, including HIV-2. - Development of thermodynamic and structural rules aimed at limiting the affinity of protease inhibitors to unwanted targets and hence improving selectivity. The goals will be achieved by a combination of experimental thermodynamic measurements (high sensitivity isothermal titration calorimetry and high sensitivity differential scanning calorimetry), structure determination (x-ray crystallography) and structure-based thermodynamic analysis. PUBLIC HEALTH RELEVANCE: More than 30 million people in the world are infected with HIV/AIDS and more than 2 million die each year. Despite their initial success, antiretroviral therapies are hindered by the emergence of drug resistant viral strains and by the occurrence of severe side effects. The main goal of this project is to develop precise guidelines for the development of antiretrovirals, especially protease inhibitors, characterized by extremely high potency, low susceptibility to drug resistance and minimal side effects.