title: PepFect 14, a novel cell-penetrating peptide for oligonucleotide delivery in solution and as solid formulation.
abstract: Numerous human genetic diseases are caused by mutations that give rise to aberrant alternative splicing. Recently, several of these debilitating disorders have been shown to be amenable for splice-correcting oligonucleotides (SCOs) that modify splicing patterns and restore the phenotype in experimental models. However, translational approaches are required to transform SCOs into usable drug products. In this study, we present a new cell-penetrating peptide, PepFect14 (PF14), which efficiently delivers SCOs to different cell models including HeLa pLuc705 and mdx mouse myotubes; a cell culture model of Duchenne's muscular dystrophy (DMD). Non-covalent PF14-SCO nanocomplexes induce splice-correction at rates higher than the commercially available lipid-based vector Lipofectamine 2000 (LF2000) and remain active in the presence of serum. Furthermore, we demonstrate the feasibility of incorporating this delivery system into solid formulations that could be suitable for several therapeutic applications. Solid dispersion technique is utilized and the formed solid formulations are as active as the freshly prepared nanocomplexes in solution even when stored at an elevated temperatures for several weeks. In contrast, LF2000 drastically loses activity after being subjected to same procedure. This shows that using PF14 is a very promising translational approach for the delivery of SCOs in different pharmaceutical forms.
authors.affiliation: Department of Neurochemistry, Stockholm University, SE-106 91 Stockholm, Sweden. kariem@neurochem.su.se
authors.forename: Kariem
authors.initials: K
authors.lastname: Ezzat
authors.forename: Samir E L
authors.initials: SE
authors.lastname: Andaloussi
authors.forename: Eman M
authors.initials: EM
authors.lastname: Zaghloul
authors.forename: Taavi
authors.initials: T
authors.lastname: Lehto
authors.forename: Staffan
authors.initials: S
authors.lastname: Lindberg
authors.forename: Pedro M D
authors.initials: PM
authors.lastname: Moreno
authors.forename: Joana R
authors.initials: JR
authors.lastname: Viola
authors.forename: Tarek
authors.initials: T
authors.lastname: Magdy
authors.forename: Rania
authors.initials: R
authors.lastname: Abdo
authors.forename: Peter
authors.initials: P
authors.lastname: Guterstam
authors.forename: Rannar
authors.initials: R
authors.lastname: Sillard
authors.forename: Suzan M
authors.initials: SM
authors.lastname: Hammond
authors.forename: Matthew J A
authors.initials: MJ
authors.lastname: Wood
authors.forename: Andrey A
authors.initials: AA
authors.lastname: Arzumanov
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gait
authors.forename: C I Edvard
authors.initials: CI
authors.lastname: Smith
authors.forename: Mattias
authors.initials: M
authors.lastname: Hällbrink
authors.forename: Ülo
authors.initials: Ü
authors.lastname: Langel
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008027:Light; D055666:Lipopeptides / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D012542:Scattering, Radiation; D012996:Solutions; D013696:Temperature
chemical_list: D057846:Cell-Penetrating Peptides; D003470:Culture Media; D016895:Culture Media, Serum-Free; D055666:Lipopeptides; D016376:Oligonucleotides, Antisense; C560878:PepFect14 peptide; D012996:Solutions