Abstract
Lipid nanoparticles (LNPs) are among the most successful classes of nonviral delivery systems for nucleic acid-based therapeutics in treating human diseases. One of the key challenges in achieving efficient cytosolic delivery of nucleic acids is overcoming endosomal entrapment within cells. Conventional lipid bilayer-forming cationic and amino lipids mediate endosomal escape via the mechanism of lamellar-to-inverted hexagonal phase transition, resulting in suboptimal cytosolic cargo delivery. pH-sensitive amphiphilic cell membrane disruption and endosomal escape have emerged as a strategy for designing protonatable or ionizable lipids, especially nonlamellar lipids, for efficient cytosolic nucleic acid delivery. Nonlamellar amino lipids possess a large wedge-shaped tail structure and do not form stable lipid bilayers. These lipids and their corresponding LNPs remain neutral, non-amphiphilic, or minimally amphiphilic at physiological pH (7.4). They become amphiphilic upon protonation or ionization in acidic endosomes (pH 6.5-5.4). The electrostatic interaction of ionized nonlamellar lipids with the negatively charged endosome membrane, combined with their large wedge-like structures, disrupts the lipid bilayer, facilitating efficient endosomal escape. Additionally, the nonlamellar ionizable lipids can be fine-tuned by altering the structure of amino head groups and lipid tails to achieve the precisely controlled pH-sensitive amphiphilic membrane disruption at endosomal pH. Therefore, these lipids exhibit excellent safety profiles and high efficiency for in vivo delivery of various therapeutic nucleic acids. pH-sensitive amphiphilic membrane disruption and endosomal escape provide a feasible and effective mechanism for designing ionizable lipids for safe and efficient in vivo nucleic acid delivery.