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High-performance liquid chromatographic determination of metallochlorophylls used for tumor phototherapy

高效液相色谱法测定肿瘤光疗用金属叶绿素

基本信息

批准号：
06453071
负责人：
INOUE Hidenari
金额：
$ 3.33万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

A series of water soluble metallochlorophylls which can be used for photosensitizers of photodynamic tumor therpay have been synthesized and determined by highperformance liquid chromatography (HPLC). Some of water soluble metallochlorophyllse. g. sodium copper (II) or iron (III) chlorophyllin, contain certain amounts of impurities, because they are usually perpared from native chlorophylls. In the present study, a preparative method for the component of metallochlorophyllins, i. e. metallopheophorbides, metallochlorin e_6, metallochlorin e_4 and metallorhodin g_7, has been developed. A reversed-phase HPLC method for their determination has been established by using a chemically bonded octadecyl silica column and an eluent of methanol-water containing phosphate buffer or 1% acetic acid. The development of the preparative and analytical methods of water soluble metallochlorophyllins has made it possible to practically examine which component is more effective for photosensitizers of tumor phototherapy. The spectroscopic properties of metallochlorophylls, e. g. electronic spectra, ESR,^<57>Fe Mossbauer spectra, CD,MCD,FAB-Mass spectra etc. have been interpreted in connection with their molecular structure to propose a guideline of the molecular design of new photosensitizers. The above spectroscopic information on water soluble metallochlorophylls would be useful in understanding the accumulation mechanism of photosensitizers on tumor cells and in designing a new photosensitizer more effective for photodynamic tumore therapy.

本文合成了一系列水溶性金属叶绿素类光动力学肿瘤治疗光敏剂，并用高效液相色谱法测定了它们的含量。一些水溶性金属叶绿素。G.叶绿素铜（II）钠或铁（III）钠含有一定量杂质，因为它们通常是天然叶绿素的过氧化物。本文介绍了一种金属叶绿素类化合物的制备方法。e.金属脱镁叶绿酸盐，金属二氢卟酚E_6，金属二氢卟酚E_4和金属紫红质G_7已被开发。采用十八烷基硅胶键合色谱柱，以甲醇-水（含磷酸盐缓冲液或1%乙酸）为流动相，建立了反相高效液相色谱法测定其含量。水溶性金属叶绿素的制备和分析方法的发展使得实际检查哪种组分对于肿瘤光疗的光敏剂更有效成为可能。研究了金属叶绿素的光谱性质。G.电子能谱、ESR、~ 1Fe<57>穆斯堡尔谱、CD、MCD、FAB-质谱等与其分子结构的关系进行了解释，为新型光敏剂的分子设计提供了指导。上述水溶性金属叶绿素的光谱信息将有助于理解光敏剂在肿瘤细胞上的积累机制，并有助于设计更有效的新型光敏剂用于光动力学肿瘤治疗。