The use of design of experiments with multiple responses to determine optimal formulations for in vivo hepatic mRNA delivery

Akari Hashiba; Manaya Toyooka; Yusuke Sato; Masatoshi Maeki; Manabu Tokeshi; Hideyoshi Harashima

doi:10.1016/j.jconrel.2020.08.031

The use of design of experiments with multiple responses to determine optimal formulations for in vivo hepatic mRNA delivery

J Control Release. 2020 Nov 10:327:467-476. doi: 10.1016/j.jconrel.2020.08.031. Epub 2020 Aug 25.

Authors

Akari Hashiba¹, Manaya Toyooka¹, Yusuke Sato², Masatoshi Maeki³, Manabu Tokeshi⁴, Hideyoshi Harashima⁵

Affiliations

¹ Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-Ku, Sapporo 060-0812, Japan.
² Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-Ku, Sapporo 060-0812, Japan. Electronic address: y_sato@pharm.hokudai.ac.jp.
³ Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita-13, Nishi-8, Kita-Ku, Sapporo 060-8628, Japan; JST PRESTO, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
⁴ Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita-13, Nishi-8, Kita-Ku, Sapporo 060-8628, Japan; Innovative Research Center for Preventive Medical Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan; Institute of Nano-Life Systems, Institutes of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
⁵ Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-Ku, Sapporo 060-0812, Japan. Electronic address: harasima@pharm.hokudai.ac.jp.

PMID: 32853732
DOI: 10.1016/j.jconrel.2020.08.031

Abstract

Although great advances have been made in the delivery of short RNAs by lipid nanoparticles (LNPs), the optimal formulation composition and physicochemical properties of LNPs for long RNA (including mRNA) remain unclear. In the present study, we optimized the lipid composition of liver-targeted mRNA-loaded LNPs that were prepared with pH-sensitive cationic lipids that had been previously designed for siRNA delivery through a two stepped design of experiment (DoE). Multiple responses including physicochemical properties, gene expression, and liver-specificity were analyzed in order, not only to understand the role of each formulation parameter, but also to examine parameters that would be difficult to predict. We found that particle size and the PEG-to-phospholipid (PEG/PL) ratio were additional key factors for liver-specific gene expression in addition to the other formulation factors. The optimized formulation showed a better gene expression compared to other lipid formulations from industry leaders. These findings suggest that a "DoE with multiple responses" approach can be used to predict significant parameters and permit optimized formulations to be prepared more efficiently.

Keywords: Design of Experiment; Lipid nanoparticles; Microfluidic device; Multiple responses; mRNA delivery.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Lipids*
Liver
Nanoparticles*
RNA, Messenger
RNA, Small Interfering

Substances

Lipids
RNA, Messenger
RNA, Small Interfering