Im Jungkyun, Das Sanket, Jeong Dongjun, Kim Chang-Jin, Lim Hyun-Suk, Kim Ki Hean, Chung Sung-Kee
Department of Chemical Engineering, Soonchunhyang University, 22 Soonchunhyangro, Asan, 31538, Republic of Korea.
Department of Chemistry, Pohang University of Science and Technology, 77 Cheongamro, Pohang, 37673, Republic of Korea.
Int J Pharm. 2017 Aug 7;528(1-2):646-654. doi: 10.1016/j.ijpharm.2017.06.030. Epub 2017 Jun 17.
The impermeability of the cell plasma membrane is one of the major barriers for protein transduction into mammalian cells, and it also limits the use of proteins as therapeutic agents. Protein transduction has usually been achieved based on certain invasive processes or cell penetrating peptides (CPP). Herein we report our study in which a synthetic guanidine-rich molecular carrier is used as a delivery vector for intracellular and transdermal delivery of proteins. First a sorbitol-based molecular carrier having 8 guanidine units (Sor-G8) was synthesized, and then was simply mixed with a cargo protein of varying sizes to form the non-covalent complex of carrier-cargo proteins. These ionic complexes were shown to have efficient cellular uptake properties. The optimum conditions including the molar ratio between cargo protein and carrier, and the treatment time have been defined. Several protein cargoes were successfully examined with differing sizes and molecular weights: green fluorescent protein (MW 27kDa), albumin (66kDa), concanavalin A (102kDa), and immunoglobulin G (150kDa). These non-covalent complexes were also found to have excellent transdermal penetration ability into the mouse skin. The skin penetration depth was studied histologically by light microscopy as well as two-photon microscopy thus generating a depth profile. These complexes were largely found in the epidermis and dermis layers, i.e. down to ca. 100μm depth of the mouse skin. Our synthetic Sor-G8 carrier was found to be substantially more efficient that Arg8 in both the intracellular transduction and the transdermal delivery of proteins. The mechanism of the cellular uptake of the complex was briefly studied, and the results suggested macropinocytosis.
细胞质膜的不渗透性是蛋白质转导进入哺乳动物细胞的主要障碍之一，它也限制了蛋白质作为治疗剂的应用。蛋白质转导通常是基于某些侵入性过程或细胞穿透肽（CPP）来实现的。在此，我们报告了一项研究，其中一种富含胍的合成分子载体被用作蛋白质细胞内和经皮递送的载体。首先合成了一种具有8个胍单元的基于山梨醇的分子载体（Sor-G8），然后将其与不同大小的货物蛋白简单混合，形成载体-货物蛋白的非共价复合物。这些离子复合物显示出具有高效的细胞摄取特性。已经确定了包括货物蛋白与载体之间的摩尔比以及处理时间在内的最佳条件。成功检测了几种大小和分子量不同的蛋白质货物：绿色荧光蛋白（分子量27kDa）、白蛋白（66kDa）、伴刀豆球蛋白A（102kDa）和免疫球蛋白G（150kDa）。还发现这些非共价复合物对小鼠皮肤具有出色的经皮渗透能力。通过光学显微镜和双光子显微镜对皮肤渗透深度进行了组织学研究，从而生成了深度剖面图。这些复合物主要存在于表皮和真皮层，即深入到小鼠皮肤约100μm的深度。我们合成的Sor-G8载体在蛋白质的细胞内转导和经皮递送方面都比Arg8效率高得多。简要研究了复合物的细胞摄取机制，结果表明是巨胞饮作用。
