TOPICAL MICROBICIDES & BIOLOGY OF VENEREAL PATHOGENS

性传播疾病是一个全球性的公共卫生问题。需要新的战略来抗击日益增长的流行病。一种有希望的新方法是开发局部杀微生物剂--安全有效的女性控制产品，专为阴道内使用而设计，旨在防止入口处的感染。在本申请中，我们提出了一个涉及三个项目和两个核心的杀菌剂开发的综合计划。该计划将研究三组精心挑选的化合物，以确定它们在体外和体内对重要STD病原体的有效性，评估相关的毒性特性，包括生殖道免疫毒性，并确定它们是否具有避孕作用。待研究的化合物包括：(1)我们先前已证明具有广泛活性且基本上无毒的硫化多糖和聚合物；(2)一系列具有抗菌特性和在人体上安全使用的化合物；以及(3)一组以前未作为潜在杀菌剂进行研究的新型化合物。在项目1中，Herold和Klotman博士将检测化合物的体外抗HSV和抗HIV-1活性以及细胞毒性。该项目将利用原代人类细胞：阴道、宫颈内和宫颈外细胞以及巨噬细胞和树突状细胞来研究抗病毒活性、作用机制和毒性。在项目2中，库珀博士将利用人类输卵管器官培养模型来确定化合物的抗衣原体和抗淋球菌活性，以研究其作用机制和毒性。项目1和2中的研究将提供关于活性光谱、作用机制和细胞毒性概况的重要信息。在这些数据的基础上，最有希望的化合物将进入项目3(Stanberry博士)的评估，其中将使用小鼠和豚鼠生殖道感染模型来研究体内对HSV和衣原体的活性。研究将调查体液(血液或精液)的影响以及与女性向男性传播性病有关的问题。可能进入临床试验的化合物将在项目3中进行检查，以确定它们是否对保护女性生殖道至关重要的先天免疫反应产生不利影响。项目1和2中的其他研究将从人类生殖道细胞和衣原体基本体中分离出硫酸乙酰肝素糖胺聚糖(GAG)，并研究特定的GAG在HSV和衣原体与细胞结合中的作用。核心A将提供行政支持。CORE B(扎内维尔德博士)将评估避孕和毒性特性(乳杆菌生长、兔阴道刺激性和急性口服毒性)，并准备配方。这一综合计划将确定和表征有前景的杀菌剂，并开发关于新化合物或新方法(GAG)的信息，这些信息将提供未来的研究领域。

Sexually transmitted diseases (STDs) are a global public health problem. New strategies are needed to combat the growing epidemic. A promising new approach is the development of topical microbicides - safe and effective, female-controlled products designed for intravaginal use and intended to prevent infection at the portal of entry. In this application we propose an integrated program for microbicide development involving three projects and two cores. The program will investigate three groups of carefully selected compounds to determine their in vitro and in vivo effectiveness against important STD pathogens, assess for pertinent toxic properties including genital tract immunotoxicity, and determine whether they are contraceptive. The compounds to be investigate include: (1) sulfated polysaccharides and polymers we have previously shown to be broadly active and essentially non-toxic; (2) a diverse collection of compounds with antimicrobial properties and a history of safe use in humans; and (3) a group of novel compounds not previously investigated as potential microbicides. In Project 1, Drs. Herold and Klotman will examine the in vitro anti-HSV and anti-HIV-1 activity and cytotoxicity of the compounds. This project will make use of primary human cells: vaginal, endocervical and ectocervical cells as well as macrophages and dendritic cells to investigate antiviral activity, mechanism of action and toxicity. In project 2, Dr. Cooper will determine the anti-chlamydial and anti-gonococcal activity of the compounds investigating mechanism of action and toxicity using the human fallopian tube organ culture model. Studies in Projects 1 and 2 will provide important information regarding the spectrum of activity, mechanism of action and cytotoxicity profiles. Based on these data, the most promising compounds will progress to evaluation in Project 3 (Dr. Stanberry), where mouse and guinea pig models of genital tract infection will be used to investigate in vivo activity against HSV and chlamydia. Studies will investigate effects of body fluids (blood or semen) and issues related to female to male STD transmission. Compounds that appear likely to enter clinical trials will be examined in Project 3 to determine whether they adversely effect innate immune responses important in protection of the female genital tract. Other studies in Projects 1 and 2 will isolate heparan sulfate glycosaminoglycans (GAGs) from human genital tract cells and chlamydial elementary bodies and investigate the role of specific GAGs in HSV and chlamydial binding to cells. Core A will provide administrative support. Core B (Dr. Zaneveld) will assess contraceptive and toxic properties (lactobacillus growth, rabbit vaginal irritation, and acute oral toxicity) and prepare formulations. This integrated program will identify and characterize promising microbicides and develop information on new compounds or approaches (GAGs) that will provide future areas for investigation.