SMALL ANIMAL MODELS FOR MICROBICIDE TESTING

该项目的总体目标是开发一种阴道给药制剂 这是一种有效的避孕方法，同时可以防止 性传播人体免疫机能丧失病毒（艾滋病毒）， 衣原体 理想情况下，这将在单一活性剂中实现，但 它的实现与兼容的组合剂可能会证明更多 现实 待测试的候选材料包括已知 干扰精子功能和被认为可能干扰的化合物 艾滋病病毒的传播机制 四种类型的体外试验 将进行：（1）细胞毒性，包括不同的试验 在不同的条件下。 (2)精子：PI。的研究， 过去25年主要集中在精子结构的研究， 功能 对精子的分析将集中在鉴定 干扰功能，包括精子代谢，运动， 宫颈粘液的渗透以及凝集剂 精子 (3)艾滋病毒：该实验室在以下方面发挥了重要作用 阐明艾滋病毒的性传播机制， 测定抑制HIV性传播的试剂的体外方法。 试验将包括基于荧光的细胞毒性和细胞间 粘附测定，以及检测试剂粘附的能力的ELISA。 干扰HIV从HIV感染的淋巴细胞向 来源于人子宫颈的上皮细胞。 (4)衣原体：筛查 抗衣原体活性将包括阻断细胞感染 来源于人类子宫颈 我们正在开发一种新技术， 衣原体检测。 该方法将采用高度敏感的 荧光细胞毒性测定并使用来源于人的细胞系 子宫颈，应该更迅速，定量，和适当的比 现有的化验。 根据文献中的信息和我们的初步发现， 选择了以下五种先导化合物进行测试：（1） 氯己定，（2）普萘洛尔，（3）4 ′-乙酰氨基苯基-4-胍基苯甲酸酯， (4)二乙基二硫代氨基甲酸酯，和（5）硫酸葡聚糖。 动物研究将 包括抑制小鼠中沙眼衣原体感染，一种系统 私家侦探曾经合作过。 此外，生育率和 将在兔子中进行阴道刺激性研究。 对于研究 猴免疫缺陷病毒（SIV）的传播，我们将合同博士。 加州灵长类动物研究中心的克里斯托弗·J·米勒说， 恒河猴中无细胞和细胞相关SIV传播的试验抑制 猴子 在五年的资助期内，我们预计将准备一个或多个 用于人体有效性临床试验的试剂。 制定 材料进行此类测试时，将特别注意 必须保护整个阴道表面，以防止 艾滋病毒的传播。 还将注意到 有效期至少24小时。 测试有效性的准备 将包括阴道刺激评估，初步 通过致突变试验评估致癌潜力，以及 在动物模型中进行致畸潜力和毒性试验。

The overall goal of this project is to develop a vaginal dosage formulation that is an effective contraceptive which simultaneously protects against the sexual transmission of human immunodeficiency virus (HIV) and Chlamydia. Ideally, this would be achieved in a single active agent, but its attainment with a compatible combination of agents may prove more realistic. Candidate materials to be tested include compounds known to interfere with sperm function and compounds considered likely to interfere with the mechanisms of transmission of HIV. Four types of in vitro tests will be carried out: (1) Cytotoxicity which will include different assays under various conditions. (2) Spermatozoa: The P.I.'s research during the last 25 years has concentrated primarily on studies of sperm structure and function. The assays of sperm will concentrate on identifying agents which interfere with function including sperm metabolism, motility, and penetration of cervical mucus as well as agents which agglutinate spermatozoa. (3) HIV: This laboratory has been instrumental in elucidating the mechanisms of sexual transmission of HIV and developing in vitro methods to assay agents which inhibit sexual transmission of HIV. Assays will include a fluorescence-based cytotoxicity and cell-cell adherence assays, and an ELISA which detects the ability of agents to interfere with transmission of HIV from HIV-infected lymphocytes to epithelial cells derived from the human cervix. (4) Chlamydia: Screening for anti-Chlamydial activity will involve blocking infection of cells derived from the human cervix. We are developing a novel technique for Chlamydial testing. This method, which will employ a highly sensitive fluorescence cytotoxicity assay and uses a cell line derived from the human cervix, should be much more rapid, quantitative, and appropriate than the existing assays. Based on information in the literature and on our preliminary findings, the following five lead compounds have been selected for testing: (1) chlorhexidine, (2) propranolol, (3) 4'-acetamidophenyl-4-guanidinobenzoate, (4) diethyldithiocarbamate, and (5) dextran sulfate. Animal studies will include inhibition of Chlamydia trachomatis infection in mice, a system which the P.I. has worked with previously. In addition, fertility and vaginal irritability studies will be carried out in rabbits. For studies of simian immunodeficiency virus (SIV) transmission we will contract Dr. Christopher J. Miller, at the California Primate Research Center, who will test inhibition of cell-free and cell-associated SIV transmission in rhesus monkeys. Within the five-year funding period we expect to have prepared one or more agents for clinical tests of effectiveness in humans. In formulating materials for such tests, particular cognizance will be taken of the necessity of protecting the entire vaginal surface in order to prevent transmission of HIV. Attention will also be directed to the desirability of effectiveness for at least 24 h. Preparation for testing effectiveness in humans will include assessment of vaginal irritation, preliminary assessment of carcinogenic potential by means of mutagenicity assays, and tests for teratogenic potential and toxicity in animal models.