IMMUNOASSAYS FOR DIDEMNINS

该项目的总体目标是开发能够 检测纳克级的敌敌畏。滴滴涕B是最有效的 抗肿瘤、抗病毒和细胞毒作用的五环家族成员 从海洋被毛类动物中分离出的脱脂肽。这个化合物目前在 作为一种抗肿瘤药物进行临床试验。现有的生物测定和高效液相色谱方法 用于检测生物基质中的敌百虫缺乏精密度和 药代动力学和药物代谢研究所需的灵敏度。我们 计划合成具有良好特性的白粉菌素A衍生免疫原 应该会引起抗体反应。这将通过耦合来实现 通过其N-甲基-D-亮氨酸侧链作用于免疫原性载体 揭示所有双异体蛋白共有的大环表位。早先的尝试 其他研究人员未能开发出放射免疫分析，可能是 由于白粉菌素B具有较强的免疫抑制作用， 由于白粉菌素A的效力弱于白粉菌素B，因此仔细操作 免疫参数，如免疫原载体的选择、免疫 免疫剂量、免疫途径、佐剂类型和预先接种 免疫原性载体，有望引起体液免疫反应。第一阶段 努力将重点放在展示开发仿制药的可行性上 所有白粉菌素的免疫分析。第二阶段将专注于提炼仿制药 测定和发展选择性免疫分析法检测毛滴虫蛋白。

The overall goal of this project is to develop immunoassays capable of detecting didemnins at the nanogram level. Didemnin B is the most potent antitumor, antiviral and cytotoxic member of a family of five cyclic depsipeptides isolated from marine tunicates. This compound is currently in clinical trials as an antitumor agent. Available bioassay and HPLC methods for detecting didemnins in biological matrices lack the precision and sensitivity required for pharmacokinetic and drug metabolism studies. We plan to synthesize well characterized didemnin A derived immunogens which should elicit antibody responses. This will be achieved by coupling didemnin A to immunogenic carriers via its N-methyl-D-leucine side chain to expose the macrocyclic epitope common among all didemnins. Earlier attempts by other investigators failed to develop a radioimmunoassay, possibly because of the strong immunosuppressive action of didemnin B. However, since didemnin A is less potent than didemnin B, careful manipulation of immunization parameters such as choice of immunogenic carrier, immunizing dose, route of immunization, adjuvant type, and prior priming with immunogenic carrier, is expected to evoke humoral immune responses. Phase I efforts will focus on demonstrating the feasibility of developing a generic immunoassay for all didemnins. Phase II will focus on refining the generic assay and develop selective immunoassays for didemnins.