Wei Yuping, Zhang Caiying, Zhang Man, Niu Qionghong, Hui Fengli, Liu Zi, Xu Xia
School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang, Henan Province, 473061, P.R. China; State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P.R. China.
School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang, Henan Province, 473061, P.R. China.
Eur J Pharm Sci. 2021 Jun 1;161:105790. doi: 10.1016/j.ejps.2021.105790. Epub 2021 Mar 6.
In our previous study, a novel cell penetrating peptide (CPP) R7 (Arg-Arg-Arg-Arg-Arg-Trp-Trp, RRRRRWW) has been developed to help cellular internalization of paclitaxel (PTX) through the non-covalent interaction with CPP. However, the facilitation mechanism of R7 mediated PTX translocation is not clear. Here the uptake pathways of R7 and R7-PTX were investigated by in vitro test and molecular simulations. In vitro experiments reveal that both R7 and R7-PTX complex translocate through the direct translocation and clathrin mediated endocytosis and associate with the macropinocytosis pathway at high CPP concentration. The translocation of R7(0.1 mM)-PTX complex further involves the lipid raft/caveolae mediated endocytosis. The simulation results show that the synergistic effect between R7 and PTX not only changes the penetration energy barrier but also activates the macropinocytosis and lipid raft/caveolae mediated pathway, resulting in the improvement in the translocation. The presence of heparin also improves the R7 and R7-PTX translocation. These studies provide a theoretical basis for understanding PTX delivery facilitated by the synergistic effect between CPP and cargo and paves a way for CPP design.
在我们之前的研究中，已开发出一种新型细胞穿透肽（CPP）R7（精氨酸-精氨酸-精氨酸-精氨酸-精氨酸-色氨酸-色氨酸，RRRRRWW），以通过与CPP的非共价相互作用帮助紫杉醇（PTX）的细胞内化。然而，R7介导的PTX转运促进机制尚不清楚。在此，通过体外试验和分子模拟研究了R7和R7-PTX的摄取途径。体外实验表明，R7和R7-PTX复合物均通过直接转运和网格蛋白介导的内吞作用进行转运，并在高CPP浓度下与巨胞饮途径相关。R7（0.1 mM）-PTX复合物的转运进一步涉及脂筏/小窝介导的内吞作用。模拟结果表明，R7和PTX之间的协同作用不仅改变了穿透能垒，还激活了巨胞饮作用和脂筏/小窝介导的途径，从而导致转运的改善。肝素的存在也改善了R7和R7-PTX的转运。这些研究为理解CPP与货物之间的协同作用促进PTX递送提供了理论基础，并为CPP设计铺平了道路。
