SMALL ANIMAL MODELS FOR MICROBICIDE TESTING

用于杀菌剂测试的小动物模型

• 批准号: 6577265
• 负责人: David Phillips
• 金额: $ 7.41万
• 依托单位: POPULATION COUNCIL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6577265
• 关键词: AIDS education /prevention; Chlamydiaceae; HIV infections; antibiotics; antiinfective agents; chemoprevention; contraceptives; cooperative study; cytotoxicity; disease /disorder prevention /control; drug adverse effect; drug screening /evaluation; epithelium; female; guinea pigs; human T cell lymphotropic virus type 1; laboratory mouse; laboratory rabbit; polysaccharides; progestins; sexually transmitted diseases; spermicide; vagina

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.

该项目的总体目标是开发阴道剂量 这是一种有效的避孕药，同时保护 人类免疫缺陷病毒（HIV）和 衣原体。 理想情况下，这将在单个活性代理中实现，但是 它与代理的兼容组合的实现可能会更多 实际的。 要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 具有艾滋病毒传播的机制。 四种体外测试 将进行：（1）细胞毒性，其中包括不同的测定 在各种条件下。 （2）精子：P.I.的研究 最近25年主要集中于精子结构的研究和 功能。 精子的测定将集中于识别剂 干扰功能，包括精子代谢，运动性和 宫颈粘液的穿透以及凝集的剂 精子。 （3）艾滋病毒：该实验室在 阐明艾滋病毒性传播和发展的机制 测定抑制HIV性传播的药物的体外方法。 测定将包括基于荧光的细胞毒性和细胞细胞 依从性测定和ELISA检测代理的能力 干扰HIV从HIV感染的淋巴细胞传播到 源自人宫颈的上皮细胞。 （4）衣原体：筛查 对于抗卫生活性，将涉及阻断细胞的感染 源自人类子宫颈。 我们正在开发一种新颖的技术 衣原体测试。 这种方法将采用高度敏感的 荧光细胞毒性测定法，并使用源自人的细胞系 子宫颈，应比 现有测定。 基于文献中的信息以及我们的初步发现 已选择以下五种铅化合物进行测试：（1） 氯己胺，（2）普萘洛尔，（3）4'-乙酰氨基苯基-4-瓜烷替苯甲酸酯， （4）二硫代氨基甲酸酯和（5）硫酸葡萄糖。 动物研究将 包括抑制小鼠的沙眼衣原体感染，一个系统 P.I.以前曾与之合作过。 另外，生育和 阴道烦躁的研究将在兔子中进行。 进行研究 Simian免疫缺陷病毒（SIV）的传播我们将收缩Dr. 加利福尼亚灵长类动物研究中心的克里斯托弗·J·米勒（Christopher J. Miller） 测试无细胞和与细胞相关的SIV传播的抑制作用 猴子。 在五年的资金期内，我们预计将准备一个或多个 对人类有效性的临床测试的代理。 在配方中 进行此类测试的材料，将采用特定的认知 必须保护整个阴道表面以防止 艾滋病毒的传播。 注意也将指向可取性 至少24小时的有效性。 准备测试有效性 在人类中，将包括对阴道刺激的评估 通过诱变性测定评估致癌潜力，并 测试动物模型中致变性潜力和毒性的测试。