Institut für Chemie und Biochemie, Freie Universität Berlin , Takustr. 3, D-14195 Berlin, Germany.
Biolitec Research GmbH , Otto-Schott-Str. 15, D-07745 Jena, Germany.
Biomacromolecules. 2018 Jan 8;19(1):222-238. doi: 10.1021/acs.biomac.7b01485. Epub 2017 Dec 12.
An adaptable approach toward cleavable nanoparticle carrier systems for photodynamic therapy (PDT) is presented, comprising a biocompatible carrier loaded with multiple photosensitizer (PS) molecules related to the clinically employed PS Temoporfin, two linkers cleavable under different triggers and glyco-targeting with mannose. A synthetic pathway to stimuli responsive hyperbranched polyglycerol (hPG) porphyrin conjugates via the copper(I)-catalyzed 1,3-dipolar cycloaddition (CuAAC) or the strain-promoted alkyne-azide cycloaddition (SPAAC) has been developed. The PS 10,15,20-tris(3-hydroxyphenyl)-5-(2,3,4,5,6-pentafluorophenyl)porphyrin was functionalized with disulfide containing cystamine and acid-labile benzacetal linkers. Conjugates with reductively and pH labile linkers were thus obtained. Cleavage of the active PS agents from the polymer carrier is shown in several different release studies. The uptake of the conjugates into the cells is demonstrated via confocal laser scanning microscopy (CLSM) and flow cytometry. Finally, the antitumor and antibacterial phototoxicity of selected conjugates has been assessed in four different tumor cell lines and in cultures of the bacterium Staphylococcus aureus. The conjugates exhibited phototoxicity in several tumor cell lines in which conjugates with reductively cleavable linkers were more efficient compared to conjugates with acid-cleavable linkers. For S. aureus, strong phototoxicity was observed for a combination of the reductively cleavable and the pH labile linker and likewise for the cleavable conjugate with mannose targeting groups. The results thus suggest that the conjugates have potential for antitumor as well as antibacterial PDT.
本文提出了一种可适应的光动力治疗(PDT)可裂解纳米颗粒载体系统的方法,该系统包含负载有多个人体细胞相关的光敏剂(PS)分子的生物相容性载体,这些 PS 分子与临床应用的 PS Temoporfin 有关,两种在不同触发条件下可裂解的连接子以及甘露糖靶向糖基化。已经开发了一种通过铜(I)催化的 1,3-偶极环加成(CuAAC)或应变促进的炔烃-叠氮化物环加成(SPAAC)来合成刺激响应超支化聚甘油(hPG)卟啉缀合物的途径。PS 10,15,20-三(3-羟基苯基)-5-(2,3,4,5,6-五氟苯基)卟啉用含二硫键的半胱胺和酸不稳定的苯乙缩醛连接子官能化。因此获得了具有还原和 pH 不稳定连接子的缀合物。在几项不同的释放研究中显示了从聚合物载体上裂解活性 PS 试剂。通过共聚焦激光扫描显微镜(CLSM)和流式细胞术证明了缀合物进入细胞的摄取。最后,在四种不同的肿瘤细胞系和金黄色葡萄球菌的培养物中评估了选定的缀合物的抗肿瘤和抗细菌光毒性。在几种肿瘤细胞系中,具有还原可裂解连接子的缀合物比具有酸可裂解连接子的缀合物更有效,表现出光毒性。对于金黄色葡萄球菌,观察到具有还原可裂解和 pH 不稳定连接子的组合以及具有甘露糖靶向基团的可裂解缀合物具有很强的光毒性。因此,结果表明缀合物具有用于抗肿瘤和抗细菌 PDT 的潜力。