Improved method for synthesis of low molecular weight protamine–siRNA conjugate

Acta Pharmaceutica Sinica B

Ergang Liu¹  Junxiao Ye²  Seung Jin Lee³  Jianxin Wang⁴  Yongzhuo Huang⁵  Zhili Yu⁶  Lu Sun⁶  Xing Pei⁶  Huining He⁶ 

[1] Collaborative Innovation Center of Chemical Science and Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China;College of Pharmacy, Tsinghua University, Beijing 100084, China;Department of Pharmacy, Ewha Womans University, Seodaemun-gu, Seoul 120-750, Republic of Korea;Key Laboratory of Smart Drug Delivery, Ministry of Education & PLA, Shanghai 201201, China;Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China;Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China;

关键词: Cell penetrating peptide;    siRNA;    Conjugate;    Conjugation yield;    Biomimetic delivery;    Crosslinker;   

DOI  :  10.1016/j.apsb.2017.11.011

来源: DOAJ

【 摘 要 】

RNAi technology has aroused wide public interest due to its high efficiency and specificity to treat multiple types of diseases. However, the effective delivery of siRNA remains a challenge due to its large molecular weight and strong anionic charge. Considering their remarkable functions in vivo and features that are often desired in drug delivery carriers, biomimetic systems for siRNA delivery become an effective and promising strategy. Based on this, covalent attachment of synthetic cell penetrating peptides (CPP) to siRNA has become of great interest. We developed a monomeric covalent conjugate of low molecular weight protamine (LMWP, a well-established CPP) and siRNA via a cytosol-cleavable disulfide linkage using PEG as a crosslinker. Results showed that the conjugates didn't generate coagulation, and exhibited much better RNAi potency and intracellular delivery compared with the conventional charge-complexed CPP/siRNA aggregates. Three different synthetic and purification methods were compared in order to optimize synthesis efficiency and product yield. The methodology using hetero-bifunctional NHS–PEG–OPSS as a crosslinker to synthesize LMWP–siRNA simplified the synthesis and purification process and produced the highest yield. These results pave the way towards siRNA biomimetic delivery and future clinical translation.

【 授权许可】

Unknown