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Attempted synthesis of taxol utilizing anti-taxol monoclonal antibody as a as a chiral mould

以抗紫杉醇单克隆抗体为手性模型尝试合成紫杉醇

基本信息

批准号：
19550114
负责人：
UESATO Shinichi
金额：
$ 3万
依托单位：
Kansai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

项目摘要

2'-Succinyltaxol-BSA and 2'-succinyltaxol-RSA conjugates were prepared. Formation of a linkage between hapten and protein is usually confirmed by the UV or fluorescamine method. However, it was difficult to confirm the binding of 2'-succinyltaxol to the protein by these methods owing to the similar UV absorption maxima of 2'-succinyltaxol and protein. We therefore conducted a mass spectrometric analysis using the neutral loss scan and MS/MS techniques to confirm the formulation of the 2'-succinyltaxol-protein conjugate. Balb/c mice were immunized i.c. at 2-week intervals each with 2'-Succinyltaxol-RSA. The spleen cells from the mice were treated in a conventional way, giving four hybridomas producing an anti-taxol monoclonal antibody. However, the hybridomas did not have a high production ability of an anti-taxol monoclonal antibody, and their antibody titers were rather low. Unfortunately, production of the antibody declined gradually with cloning. This problem was not improved in spite of our efforts with amounts of antibody and kinds of adjuvant as well as culturing conditions. Condensation reaction between baccacin III and an activated ester (viniyl ester) of C13-carboxylic acid was attempted under a crude antibody as a preliminary experiment. Further examination of the antibody reaction has not yet been conducted owing to a loss of antibody production function.

制备了2 ′-琥珀酰紫杉醇-BSA和2 ′-琥珀酰紫杉醇-RSA偶联物。半抗原和蛋白质之间的连接的形成通常通过UV或荧光胺法来确认。然而，由于2 '-琥珀酰紫杉醇和蛋白质的紫外吸收最大值相似，因此难以通过这些方法确认2'-琥珀酰紫杉醇与蛋白质的结合。因此，我们使用中性丢失扫描和MS/MS技术进行质谱分析，以确认2 '-琥珀酰紫杉醇-蛋白质缀合物的制剂。将Balb/c小鼠i.c.每隔2周用2 '-琥珀酰紫杉醇-RSA。以常规方式处理小鼠的脾细胞，得到四个产生抗紫杉醇单克隆抗体的杂交瘤。然而，杂交瘤不具有抗紫杉醇单克隆抗体的高生产能力，并且它们的抗体滴度相当低。不幸的是，随着克隆的进行，抗体的产量逐渐下降。尽管我们在抗体量、佐剂种类以及培养条件方面进行了努力，但这一问题并未得到改善。作为初步实验，在粗制抗体下尝试了巴卡星III与C13-羧酸的活化酯（乙烯基酯）之间的缩合反应。由于抗体产生功能丧失，尚未对抗体反应进行进一步检查。