Unraveling membrane perturbation mechanisms and developing novel evaluation methods for controlling the function of cell-penetrating peptides

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K15982
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
• Research Institution: National Institute of Health Sciences
• Principal Investigator: Haraya Yuki 国立医薬品食品衛生研究所, 薬品部, 主任研究官 (30634604)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 細胞膜透過ペプチド / 脂質膜 / 膜摂動 / 膜透過 / 原子間力顕微鏡法 / ドラッグデリバリー

Outline of Final Research Achievements

We hypothesized that cell-penetrating peptides, which enable intracellular delivery of drug molecules, exert their cell membrane penetration through mechanical perturbation on plasma lipid membranes. We defined the effect of peptides that reduce the membrane stiffness of artificial cell membranes (liposomes) as 'membrane perturbation' and developed an atomic force microscopy method to directly quantify it. This allowed for quantitative evaluation of the relationship between the membrane perturbation of various peptides with different amino acid sequences and structural conformations, and their cell membrane penetration and cytotoxicity. As a result, we demonstrated a novel methodology for designing peptides with high cell penetration ability.

Free Research Field

物理系薬学/生物物理化学

Academic Significance and Societal Importance of the Research Achievements

本研究成果は，「細胞毒性が低くかつ高い細胞膜透過性を示すペプチドを開発するためには，膜摂動力を最適化する必要がある」という新たな学術的仮説を提示すると同時に，この方法論の実践のために構築した評価法の有用性を示すものである．本評価法を用いて，塩基性アミノ酸/疎水性アミノ酸数，疎水性度，両親媒性度などの構造特性を説明変数とする様々なペプチドの膜摂動力と機能の関係について解明を進めることで，より有用な薬物キャリアの開発が可能となる．また，膜摂動力と細胞毒性の連関解明に本評価手法を用いることで，ペプチドの細胞膜障害性を利用した「耐性のつきにくい抗がん/抗菌剤」の開発に資する基盤研究を展開できる．