Bioconjugatable porphyrins bearing a compact swallowtail motif for water solubility
Bioconjugatable porphyrins bearing a compact swallowtail motif for water solubility
复制标题
DOI：
10.1021/bc050337w
复制
发表时间：
2006-05-17
影响因子：
4.7
通讯作者：
Lindsey, Jonathan S.
中科院分区：
文献类型：
--
作者：
Borbas, K. Eszter;Mroz, Pawel;Lindsey, Jonathan S.
广泛的应用需要获得水溶性、可生物共轭的卟啉。已知在支化位点连接至卟啉内消旋位置的支化烷基可赋予高有机溶解度。这种在每个分支末端带有极性基团（羟基、二羟基磷酰基、二羟基磷酰氧基）的“燕尾”基序现已被掺入反式-AB-卟啉的内消旋位点。燕尾基序的掺入依赖于合理的合成方法，其中1,9-双(N-丙基亚氨基)二吡咯甲烷（在5位带有可生物共轭的系链）与二吡咯甲烷（在5位带有受保护的1,5-二羟基戊-3-基单元）缩合。所得锌卟啉的燕尾基序中的两个羟基可以转化为相应的二磷酸盐或二膦酸盐产物。 4-(羧甲基氧基)苯基提供生物可缀合的系链。通过目视检查、紫外-可见吸收光谱或 H-1 NMR 光谱测定，本文报道的六种此类卟啉具有高度水溶性（在室温下，在 pH 7 的水中，>= 20 mM）。在水溶液中与未保护的卟啉二磷酸酯和针对 T 细胞受体 CD3 ε 的单克隆抗体进行共价连接。在流式细胞术和荧光显微镜测定中，所得缀合物与 Jurkat 细胞的市售异硫氰酸荧光素标记抗体的性能相当。总而言之，这项工作能够以紧凑的结构制备有用数量的水溶性、可生物共轭的卟啉，用于生命科学的应用。
A broad range of applications requires access to water-soluble, bioconjugatable porphyrins. Branched alkyl groups attached at the branching site to the porphyrin meso position are known to impart high organic solubility. Such "swallowtail" motifs bearing a polar group (hydroxy, dihydroxyphosphoryl, dihydroxyphosphoryloxy) at the terminus of each branch have now been incorporated at a meso site in trans-AB-porphyrins. The incorporation of the swallowtail motif relies on rational synthetic methods whereby a 1,9-bis(N-propylimino) dipyrromethane (bearing a bioconjugatable tether at the 5-position) is condensed with a dipyrromethane (bearing a protected 1,5-dihydroxypent-3-yl unit at the 5-position). The two hydroxy groups in the swallowtail motif of each of the resulting zinc porphyrins can be transformed to the corresponding diphosphate or diphosphonate product. A 4-(carboxymethyloxy) phenyl group provides the bioconjugatable tether. The six such porphyrins reported here are highly water-soluble (>= 20 mM at room temperature in water at pH 7) as determined by visual inspection, UV-vis absorption spectroscopy, or H-1 NMR spectroscopy. Covalent attachment was carried out in aqueous solution with the unprotected porphyrin diphosphonate and a monoclonal antibody against the T-cell receptor CD3 epsilon. The resulting conjugate performed comparably to a commercially available fluorescein isothiocyanate-labeled antibody with Jurkat cells in flow cytometry and fluorescence microscopy assays. Taken together, this work enables preparation of useful quantities of water-soluble, bioconjugatable porphyrins in a compact architecture for applications in the life sciences.