Design and Synthesis of New Cell Penetrating Peptides: Diffusion and Distribution Inside the Cornea.
The role of cell penetrating peptides (CPPs) has been challenged in recent years for drug delivery to ocular tissues for the targeting of both anterior and posterior segments. The enhancement of trans-corneal transport for anterior segment targeting is a very important issue possibly leading to important outcomes on efficacy and to the opportunity of topical administration of molecules with unfavorable penetration properties. The aim of the present work was the design and synthesis of new CPPs, deriving from the structure of PEP-1 peptide. Synthesized peptides were labeled with 5-carboxyfluorescein (5-FAM), and their diffusion behavior and distribution inside the cornea were evaluated by a validated ex vivo model and a confocal microscopy approach. Newly synthesized peptides showed similar corneal permeation profiles as PEP-1 (Papp= 0.75 ± 0.56 × 10-6cm/s), about 2.6-fold higher than 5-FAM (Papp= 0.29 ± 0.08 × 10-6cm/s) despite the higher molecular weight. Confocal microscopy experiments highlighted the tendency of PEP-1 and its derived peptides to localize in the intercellular space and/or in the plasma membrane. Noteworthy, using penetratin as positive control, a higher trans-corneal permeation (Papp= 6.18 ± 1.46 × 10-6cm/s) was evidenced together with a diffusion by intracellular route and a different accumulation between wings and basal epithelial cells, probably depending on the stage of cell development. Finally, PEP-1 and pep-7 proved to be safe and well tolerated when tested on human conjuctival cell line.
Department of Pharmacy, University of Parma , Parco Area delle Scienze 27/A, 43124 Parma, Italy.
Silvia
S
Pescina
Department of Pharmacy, University of Salerno , Via G. Paolo II 132, 84084 Fisciano (SA), Italy.
Marina
M
Sala
Department of Pharmacy, University of Parma , Parco Area delle Scienze 27/A, 43124 Parma, Italy.
Cristina
C
Padula
Department of Pharmacy, University of Salerno , Via G. Paolo II 132, 84084 Fisciano (SA), Italy.
Maria Carmina
MC
Scala
Department of Pharmacy, University of Salerno , Via G. Paolo II 132, 84084 Fisciano (SA), Italy.
Antonia
A
Spensiero
Department of Biomedical, Biotechnological and Translational Sciences, University of Parma , Via Volturno 39, 43126 Parma, Italy.
Silvana
S
Belletti
Department of Biomedical, Biotechnological and Translational Sciences, University of Parma , Via Volturno 39, 43126 Parma, Italy.
Rita
R
Gatti
Department of Pharmacy, University of Naples Federico II , Via D. Montesano 49, 80131 Napoli, Italy.
Ettore
E
Novellino
Department of Pharmacy, University of Salerno , Via G. Paolo II 132, 84084 Fisciano (SA), Italy.
Pietro
P
Campiglia
Department of Pharmacy, University of Parma , Parco Area delle Scienze 27/A, 43124 Parma, Italy.
Patrizia
P
Santi
Department of Pharmacy, University of Parma , Parco Area delle Scienze 27/A, 43124 Parma, Italy.
Sara
S
Nicoli
Department of Pharmacy, University of Naples Federico II , Via D. Montesano 49, 80131 Napoli, Italy.
Carmine
C
Ostacolo
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003315:Cornea / Q000378:metabolism*; D003543:Cysteamine / Q000031:analogs & derivatives / Q000378:metabolism; D005452:Fluoresceins / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008872:Microwaves; D010455:Peptides / Q000378:metabolism; D013552:Swine
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D005452:Fluoresceins; C503924:Pep-1 peptide; D010455:Peptides; D003543:Cysteamine; C045132:4-carboxyfluorescein; C414409:penetratin
cell penetrating peptides; confocal microscopy; ocular drug delivery; penetratin; pep-1; trans-corneal delivery