TOPICAL MICROBICIDES & BIOLOGY OF VENEREAL PATHOGENS

性传播疾病（STDs）是一个全球性的公共卫生问题。 需要制定新的战略来防治这一日益严重的流行病。 一种有希望的新方法是开发局部杀微生物剂-安全有效，女性控制的产品，设计用于阴道内使用，旨在预防入口感染。 在这个应用程序中，我们提出了一个综合计划，杀微生物剂的发展，涉及三个项目和两个核心。 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的性病病原体的有效性，评估相关的毒性，包括生殖道免疫毒性，并确定它们是否是避孕药。待研究的化合物包括：（1）硫酸多糖和聚合物，我们以前已经证明是广泛的活性和基本上无毒的;（2）具有抗菌特性和人类安全使用历史的化合物的多样化集合;和（3）一组以前没有研究过的新型化合物作为潜在的杀微生物剂。在项目1中，赫罗尔德博士和克洛特曼博士将检查化合物的体外抗单纯疱疹病毒和抗艾滋病毒-1活性以及细胞毒性。 该项目将利用原代人类细胞：阴道、子宫颈内和子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，库珀博士将使用人输卵管器官培养模型确定化合物的抗衣原体和抗淋球菌活性，研究作用机制和毒性。项目1和2的研究将提供有关活性谱、作用机制和细胞毒性特征的重要信息。 基于这些数据，最有前途的化合物将在项目3（Stanberry博士）中进行评价，其中将使用小鼠和豚鼠生殖道感染模型研究体内抗HSV和衣原体活性。 研究将调查体液（血液或精液）的影响以及与女性到男性的性病传播有关的问题。 将在项目3中检查似乎可能进入临床试验的化合物，以确定它们是否对保护女性生殖道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生殖道细胞和衣原体原体中分离硫酸乙酰肝素糖胺聚糖（GAG），并研究特异性GAG在HSV和衣原体与细胞结合中的作用。 核心A将提供行政支助。核心B（Dr. Zaneveld）将评估避孕和毒性特性（阴道生长、家兔阴道刺激和急性经口毒性）并制备制剂。 该综合计划将确定和表征有前途的杀微生物剂，并开发新化合物或方法（GAGs）的信息，为未来的研究提供领域。

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.