Screening and optimization of ligand conjugates for lysosomal targeting.
Screening and optimization of ligand conjugates for lysosomal targeting.
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DOI：
10.1021/bc200336j
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发表时间：
2011-11-16
影响因子：
4.7
通讯作者：
Torchilin, Vladimir P.
中科院分区：
文献类型：
--
作者：
Meerovich, Igor;Koshkaryev, Alexander;Thekkedath, Ritesh;Torchilin, Vladimir P.
溶酶体靶向脂质体的使用可以显著改善治疗酶和伴侣蛋白进入溶酶体治疗溶酶体储存障碍的效果。本研究的目的是在中性红和罗丹明B的基础上合成新的潜在溶酶体亲和性配体，并研究它们对模型化合物异硫氰酸荧光素-葡聚糖(FD)表面修饰脂质体的特异性递送作用。用共聚焦免疫荧光显微镜、亚细胞分裂和流式细胞术研究脂质体在HeLa细胞中的转运及其在溶酶体内的含量。共聚焦显微镜显示，经罗丹明B衍生物修饰的脂质体具有良好的共定位能力，并能很好地识别特异性溶酶体标记物。亚细胞分级法分离的Fd在溶酶体中的荧光比较也表明，经部分合成配体修饰的脂质体在溶酶体中的载脂效率明显高于普通脂质体。用含有溶酶体内β-半乳糖苷酶特异性底物5-十二酰氨基荧光素二-D-半乳糖苷的脂质体处理的完整细胞的流式细胞术进一步证实了这些结果，使用了一些细胞系，包括诱导表型为溶酶体酶缺陷的巨噬细胞；两个合成的配体-罗丹明B DSPE-PEG2K-酰胺和6-(3-(DSPE-PEG2k)-硫脲)罗丹明B-表现出增强的溶酶体转运，在某些情况下，高于商用的罗丹明B十八酯，对于诱导溶酶体酶缺乏表型的细胞表现出最好的结果(与普通脂质体相比，对溶酶体输送的促进程度高达75%)。使用罗丹明B衍生物修饰的脂质体可能有利于开发用于治疗溶酶体相关疾病的药物输送系统。
The use of lysosome-targeted liposomes may significantly improve the delivery of therapeutic enzymes and chaperones into lysosomes for the treatment of lysosomal storage disorders. The aim of this research was to synthesize new potentially lysosomotropic ligands on a base of Neutral Red and rhodamine B and to study their ability to enhance specific lysosomal delivery of surface-modified liposomes loaded with a model compound, fluorescein isothiocyanate-dextran (FD). The delivery of these liposomes and their content to lysosomes in HeLa cells was investigated by confocal immunofluorescent microscopy, subcellular fractionation and flow cytometry. Confocal microscopy demonstrated that liposomes modified with derivatives of rhodamine B provide good rate of co-localization well the specific lysosomal markers. The comparison of fluorescence of FD in lysosomes isolated by subcellular fractionation also showed that the efficiency of lysosomal delivery of liposomal load by liposomes modified with some of synthesized ligands was significantly higher compared with plain liposomes. These results were additionally confirmed by the flow cytometry of the intact cells treated with liposomes loaded with with 5-dodecanoylaminofluorescein di-β-D-galactopyranoside, a specific substrate for the intralysosomal β-galactosidase, using a number of cell lines, including macrophages with induced phenotype of lysosomal enzyme deficiency; two of the synthesized ligands – rhodamine B DSPE-PEG2k-amide and 6-(3-(DSPE-PEG2k)-thioureido) rhodamine B – demonstrated enhanced lysosomal delivery, in some cases, higher than that for commercially available rhodamine B octadecyl ester, with the best results (the enhancement of the lysosomal delivery up to 75% greater in comparison to plain liposomes) shown for the cells with induced lysosomal enzyme deficiency phenotype. Use of liposomes modified with rhodamine B derivatives may be advantageous for the development of drug delivery systems for the treatment of lysosome-associated disorders.
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影响因子：
4.1
作者：
KUWANA, T;MULLOCK, BM;LUZIO, JP
通讯作者：
LUZIO, JP
影响因子：
10.8
作者：
Boddapati, Sarathi V.;D'Souza, Gerard G. M.;Weissig, Volkmar
通讯作者：
Weissig, Volkmar
影响因子：
6.4
作者：
SUNDSTROM, C;NILSSON, K
通讯作者：
NILSSON, K
影响因子：
0.9
作者：
Derkacheva, V. M.;Mikhalenko, S. A.;Luk'yanets, E. A.
通讯作者：
Luk'yanets, E. A.
影响因子：
2.5
作者：
Minier, C;Moore, MN
通讯作者：
Moore, MN