D-SAP: a new, noncytotoxic, and fully protease resistant cell-penetrating peptide.
Protease resistant cell-penetrating peptides (CPPs) are promising carriers for drugs unable to cross the cell membrane. As these CPPs are stable in vivo for much longer periods of time compared to other classes of therapeutic peptides, noncytotoxicity is a property sine qua non for their pharmacological development. Described herein is a fully protease resistant CPP that is noncytotoxic at concentrations up to 1 mM. Proteolytic stability was obtained by chiral inversion of the residues of a known self-assembling CPP-from all L-amino acids to all D-amino acids-and then assessed against trypsin and human serum. Circular dichroism studies confirmed the enantiomeric structure of the analogue, and transmission electron microscopy (TEM) studies indicated that the new inverso analogue retains the ability of the original peptide to self-assemble. The results of uptake experiments indicate that the protease-stable (that is, D-amino acid) analogue of the peptide is internalised by cells to the same extent as the protease-susceptible (that is, L-amino acid) parent peptide. Also reported herein are the results of studies on the cellular internalisation mechanism of the all-D analogue, which reveal the steps followed by the peptide upon its entry into the cell.
Institut de Recerca Biomèdica, Parc Científic de Barcelona, Josep Samitier 1-5, 08028 Barcelona, Spain.
Sílvia
S
Pujals
Jimena
J
Fernández-Carneado
M Dolors
MD
Ludevid
Ernest
E
Giralt
D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002942:Circular Dichroism; D004337:Drug Carriers / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D049231:Saposins / Q000737:chemistry / Q000378:metabolism*
D004337:Drug Carriers; D010455:Peptides; D049231:Saposins; D010450:Endopeptidases