title: (31)P solid-state NMR based monitoring of permeation of cell penetrating peptides into skin.
abstract: The main objective of the current study was to investigate penetration of cell penetrating peptides (CPPs: TAT, R8, R11, and YKA) through skin intercellular lipids using (31)P magic angle spinning (MAS) solid-state NMR. In vitro skin permeation studies were performed on rat skin, and sections (0-60, 61-120, and 121-180μm) were collected and analyzed for (31)P NMR signal. The concentration-dependent shift of 0, 25, 50, 100, and 200mg/ml of TAT on skin layers, diffusion of TAT, R8, R11, and YKA in the skin and time dependent permeation of R11 was measured on various skin sections using (31)P solid-state NMR. Further, CPPs and CPP-tagged fluorescent dye encapsulate liposomes (FLip) in skin layers were tagged using confocal microscopy. The change in (31)P NMR chemical shift was found to depend monotonically on the amount of CPP applied on skin, with saturation behavior above 100mg/ml CPP concentration. R11 and TAT caused more shift in solid-state NMR peaks compared to other peptides. Furthermore, NMR spectra showed R11 penetration up to 180μm within 30min. The results of the solid-state NMR study were in agreement with confocal microscopy studies. Thus, (31)P solid-state NMR can be used to track CPP penetration into different skin layers.
authors.affiliation: College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, USA.
authors.forename: Pinaki R
authors.initials: PR
authors.lastname: Desai
authors.affiliation: Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, USA; National High Magnetic Field Laboratory, Tallahassee, USA.
authors.forename: Ashley R
authors.initials: AR
authors.lastname: Cormier
authors.affiliation: College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, USA.
authors.forename: Punit P
authors.initials: PP
authors.lastname: Shah
authors.affiliation: Dr. Reddys Laboratories, Integrated Product Development, Hyderabad, India.
authors.forename: Ram R
authors.initials: RR
authors.lastname: Patlolla
authors.affiliation: Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, USA; National High Magnetic Field Laboratory, Tallahassee, USA. Electronic address: paravastu@eng.fsu.edu.
authors.forename: Anant K
authors.initials: AK
authors.lastname: Paravastu
authors.affiliation: College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, USA. Electronic address: mandip.sachdeva@gmail.com.
authors.forename: Mandip
authors.initials: M
authors.lastname: Singh
mesh_terms: D000818:Animals; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010539:Permeability; D051381:Rats; D054772:Rats, Hairless; D012867:Skin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D057846:Cell-Penetrating Peptides; D005456:Fluorescent Dyes; D008055:Lipids; D008081:Liposomes
keywords: Cell penetrating peptides; Confocal laser scanning microscopy; Polyarginine peptide; Skin permeation; Solid-state NMR; TAT peptide; Topical delivery