SMALL ANIMAL MODELS FOR MICROBICIDE TESTING

该项目的总体目标是开发一种阴道用药配方。 这是一种有效的避孕药，同时可以防止 人类免疫缺陷病毒(HIV)的性传播和 衣原体。理想情况下，这将在单一的活性试剂中实现，但是 通过一种相容的药物组合来实现它可能证明更多 现实主义。待测试的候选材料包括已知的化合物 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。四种体外试验 将进行：(1)细胞毒性，包括不同的检测方法 在各种情况下。(2)精子：P.I.S时期的研究 在过去的25年里，主要集中在精子结构和 功能。对精子的分析将集中在鉴定哪些物质 干扰功能，包括精子新陈代谢、活力和 宫颈粘液和凝集剂的穿透 精子。(3)艾滋病毒：这个实验室在以下方面发挥了作用 人类免疫缺陷病毒性行为传播机制的研究进展 体外方法检测抑制HIV性传播的试剂。 检测将包括基于荧光的细胞毒性和细胞-细胞 依从性试验，以及检测代理人是否有能力 干扰HIV从感染HIV的淋巴细胞传播到 来源于人类子宫颈的上皮细胞。(4)衣原体：筛查 因为抗衣原体的活性将包括阻断细胞的感染 取自人类的宫颈。我们正在开发一种新的技术来治疗 衣原体检测。这种方法，它将使用高度敏感的 荧光细胞毒性试验，并使用了一种来自人类的细胞系 宫颈，应该更快，更定量，更合适 现有的化验方法。 根据文献中的信息和我们的初步调查结果， 选择了以下五种先导化合物进行测试：(1) 氯己定，(2)普奈洛尔，(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；(5)葡聚糖硫酸盐。动物研究将会 包括抑制小鼠沙眼衣原体感染，一种系统 私家侦探之前已经处理过了。此外，生育率和 将在兔子身上进行阴道刺激性研究。用于研究 关于猴免疫缺陷病毒(SIV)的传播，我们将与Dr。 加州灵长类动物研究中心的克里斯托弗·J·米勒，他将 实验抑制恒河猴体内无细胞和细胞相关的SIV传播 猴子。 在五年的资助期内，我们预计已准备好一个或多个 用于人体疗效临床测试的试剂。在制定 对于此类测试的材料，将特别注意 有必要保护整个阴道表面，以防止 艾滋病毒的传播。还将把注意力集中在可取性上 至少24小时的有效性。为测试有效性做准备 在人类身上将包括对阴道刺激的评估，初步 通过诱变试验评估致癌潜力，以及 动物模型的致畸潜力和毒性测试。

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.