PROTEGRIN DESIGN

我们的长期目标是设计可同时用作局部杀菌剂以防止性传播疾病(STD)和局部治疗细菌性阴道病(BV)的前列腺素多肽。本项目的具体目标是：1.设计类似前列环素的分子，在不影响正常阴道菌群的情况下，灭活多种性病病原体和与细菌性阴道病相关的细菌。2.确定前列环素的β-折叠和转折区如何参与这些活动。3.了解前列环素如何与与其用作局部杀微生物剂有关的因素相互作用，包括a)宿主蛋白、多肽、多肽和细胞；b)宿主和微生物蛋白酶；c)非氧合酶-9其他表面活性剂。4.研究前列环素对阴道机会主义者白色念珠菌的影响。5.研究前胶原如何组装成二聚体和寡聚体，并确定这种组装是否以及如何与它们的抗菌、细胞毒性和溶血特性有关。前列环素是一种小的、非常有效的β-折叠多肽，可以迅速灭活许多微生物，包括那些导致大多数性传播细菌感染的微生物。我们将使用固相肽合成和精确的抗菌检测方法来“微调”前列环素，为未来的阴道应用做准备。我们的目的是开发不影响阴道乳杆菌(例如嗜酸乳杆菌和皱纹乳杆菌)，但对白色念珠菌、性传播疾病细菌和与细菌性阴道炎/阴道病相关的菌群具有高度活性的前列环素样肽。我们可以通过在蛋白质中引入一个或两个氨基酸取代来获得这一系列性质，这些蛋白质具有15-18个残基和两个分子内二硫键。我们计划产生数量相对较少的额外前胶原变异体，并测试它们对一组STD靶标生物体的活性。总体而言，这些研究将有助于开发专门为阴道内使用的新型含肽局部杀微生物剂。鉴于性传播疾病的流行和严重后果，迫切需要能够保护和增强妇女能力的局部杀微生物剂。

Our long term goal is to design protegrin peptides that will be used both as topical microbicides to prevent sexually transmitted diseases (STDs) and as topical therapeutics to remediate bacterial vaginosis (BV). The Specific Aims of this project are: 1. To design protegrin-like molecules that inactivate multiple STD agents and the bacteria associated with bacterial vaginosis, without affecting normal vaginal flora. 2. To determine how the beta-sheet and turn regions of protegrins contribute to these activities. 3. To learn how protegrins interact with factors relevant to their use as topical microbicides, including a) host proteins, peptides, peptides and cells; b) host and microbial proteases; c) nonoxynol-9 other surfactants. 4. To study the effects of protegrins on C. albicans a frequent vaginal opportunist. 5. To examine how protegrins assemble into dimers and oligomers, and ascertain if and how such assemble relates to their antimicrobial, cytotoxic and hemolytic properties. Protegrins are small, exceptionally potent, beta-sheet peptides that rapidly inactivate many microbes, including those responsible for most sexually transmitted bacterial infections. We will use solid-phase peptide synthesis and precise methods of antimicrobial testing to "fine tune" protegrins for future intravaginal application. Our intent is to develop protegrin-like peptides that do not affect vaginal lactobacilli (e.g., L. acidophilus and L. crispatus), but are highly active against C. albicans, STD bacteria, and the flora associated with bacterial vaginitis/vaginosis. We can obtain this constellation of properties by introducing one or two amino acid substitutions into protegrins with 15-18 residues and two intramolecular disulfide bonds. We plan to generate a relatively small number of additional protegrin variants and to test their activity against a panel of STD target organisms. Overall, these studies will facilitate the development of novel, peptide- containing topical microbicides that are designed specifically for intravaginal use. Given the prevalence and serious consequences of STDs, topical microbicides that can protect and empower women are urgently needed.