title: Advantages of ethanol dilution method for preparing GALA-modified liposomal siRNA carriers on the in vivo gene knockdown efficiency in pulmonary endothelium.
abstract: We previously reported that a multifunctional envelope-type nano device (MEND) modified with a GALA peptide (GALA/MEND) exerted dual functions; effective targeting the pulmonary endothelium and endosomal escape. The GALA/MEND containing encapsulated siRNA was originally prepared by the film coated hydration method (GALA/MENDHyd). However, an ethanol dilution method was found to be more appropriate for scaling up the preparation of this liposomal nanoparticle. In this study, we report on the preparation of a GALA/MEND based on the principal of the ethanol dilution (GALA/MENDEtOH). The gene knockdown efficiency of the MENDHyd and MENDEtOH without GALA-modification was equivalent regardless of the method used in the preparation. The GALA/MENDEtOH induced more efficient gene silencing in the pulmonary endothelium (ED50; approximately 0.17 mg siRNA/kg) compared to the GALA/MENDHyd. The GALA/MENDEtOH escaped from endosomes more rapidly than GALA/MENDHyd, while the pharmacokinetics and lung accumulation of GALA/MENDEtOH and GALA/MENDHyd were comparable after i.v. administration. Collectively, the ethanol dilution method improves the function of the GALA/MEND as a lung-targeting siRNA carrier.
authors.affiliation: Laboratory for Formulation research, Taiho Pharmaceutical Co., Ltd., 224-2 Ebisuno, Hiraishi, Kawauchi-cho, Tokushima 771-0194, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kusumoto
authors.affiliation: Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan. Electronic address: akita@pharm.hokudai.ac.jp.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Akita
authors.affiliation: Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Sarochin
authors.initials: S
authors.lastname: Santiwarangkool
authors.affiliation: 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.
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
mesh_terms: D000818:Animals; D004339:Drug Compounding / Q000379:methods; D004727:Endothelium / Q000378:metabolism; D000431:Ethanol / Q000737:chemistry*; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008081:Liposomes; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000008:administration & dosage / Q000737:chemistry*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013154:Spleen / Q000378:metabolism
chemical_list: D008055:Lipids; D008081:Liposomes; D010455:Peptides; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012997:Solvents; C066951:GALA peptide; D000431:Ethanol; D011094:Polyethyleneimine
keywords: Ethanol dilution method; Intracellular trafficking; Lipid nanoparticles; Lung endothelium; siRNA