Functional L-Arginine Derivative as an Efficient Vector for Intracellular Protein Delivery for Potential Cancer Therapy

Xiao He; Yannv Qu; Su Xiong; Zhiru Jiang; Yaqin Tang; Fei Yan; Yuanfei Deng; Yansun Sun

doi:10.3390/jfb14060301

Functional L-Arginine Derivative as an Efficient Vector for Intracellular Protein Delivery for Potential Cancer Therapy

J Funct Biomater. 2023 May 30;14(6):301. doi: 10.3390/jfb14060301.

Authors

Xiao He^{1

2}, Yannv Qu¹, Su Xiong³, Zhiru Jiang¹, Yaqin Tang³, Fei Yan², Yuanfei Deng¹, Yansun Sun¹

Affiliations

¹ Department of Geriatrics, Peking University Shenzhen Hospital, Shenzhen 518036, China.
² Center for Cell and Gene Circuit Design, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
³ Chongqing Key Laboratory of Medicinal Chemistry and Molecular Pharmacology, Chongqing University of Technology, Chongqing 400054, China.

Abstract

The utilization of cytosolic protein delivery is a promising approach for treating various diseases by replacing dysfunctional proteins. Despite the development of various nanoparticle-based intracellular protein delivery methods, the complicated chemical synthesis of the vector, loading efficiency and endosomal escape efficiency of proteins remain a great challenge. Recently, 9-fluorenylmethyloxycarbonyl (Fmoc)-modified amino acid derivatives have been used to self-assemble into supramolecular nanomaterials for drug delivery. However, the instability of the Fmoc group in aqueous medium restricts its application. To address this issue, the Fmoc ligand neighboring arginine was substituted for dibenzocyclooctyne (DBCO) with a similar structure to Fmoc to obtain stable DBCO-functionalized L-arginine derivative (DR). Azide-modified triethylamine (crosslinker C) was combined with DR to construct self-assembled DRC via a click chemical reaction for delivering various proteins, such as BSA and saporin (SA), into the cytosol of cells. The hyaluronic-acid-coated DRC/SA was able to not only shield the cationic toxicity, but also enhance the intracellular delivery efficiency of proteins by targeting CD44 overexpression on the cell membrane. The DRC/SA/HA exhibited higher growth inhibition efficiency and lower IC₅₀ compared to DRC/SA toward various cancer cell lines. In conclusion, DBCO-functionalized L-arginine derivative represents an excellent potential vector for protein-based cancer therapy.

Keywords: DBCO; L-arginine derivatives; click chemical reaction; protein delivery; self-assembly.

Grants and funding

This work was supported by the Shenzhen Science and Technology Program (JCYJ20210324110004012), the Science and Technology research program of Chongqing Education Commission of China (KJQN202201168) and the Scientific Research Foundation of Chongqing University of Technology.