title: Synthesis and in vitro evaluation of PNA-peptide-DETA conjugates as potential cell penetrating artificial ribonucleases.
abstract: We report the synthesis of novel artificial ribonucleases with potentially improved cellular uptake. The design of trifunctional conjugates 1a and 1b is based on the specific RNA-recognizing properties of PNA, the RNA-cleaving abilities of diethylenetriamine (DETA), and the peptide (KFF)(3)K for potential uptake into E. coli. The conjugates were assembled in a convergent synthetic route involving native chemical ligation of a PNA, containing an N-terminal cysteine, with the C-terminal thioester of the cell-penetrating (KFF)(3)K peptide to give 12a and 12b. These hybrids contained a free cysteine side-chain, which was further functionalized with an RNA-hydrolyzing diethylenetriamine (DETA) moiety. The trifunctional conjugates (1a, 1b) were evaluated for RNA-cleaving properties in vitro and showed efficient degradation of the target RNA at two major cleavage sites. It was also established that the cleavage efficiency strongly depended on the type of spacer connecting the PNA and the peptide.
authors.affiliation: Leiden Institute of Chemistry and ProSensa B.V., Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: Lene
authors.initials: L
authors.lastname: Petersen
authors.forename: Martijn C
authors.initials: MC
authors.lastname: de Koning
authors.forename: Petra
authors.initials: P
authors.lastname: van Kuik-Romeijn
authors.forename: Jimmy
authors.initials: J
authors.lastname: Weterings
authors.forename: Christine J
authors.initials: CJ
authors.lastname: Pol
authors.forename: Gerard
authors.initials: G
authors.lastname: Platenburg
authors.forename: Mark
authors.initials: M
authors.lastname: Overhand
authors.forename: Gijsbert A
authors.initials: GA
authors.lastname: van der Marel
authors.forename: Jacques H
authors.initials: JH
authors.lastname: van Boom
mesh_terms: D004926:Escherichia coli / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics; D011073:Polyamines / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics; D012313:RNA / Q000187:drug effects / Q000378:metabolism; D012260:Ribonucleases / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics; D013379:Substrate Specificity
chemical_list: D020135:Peptide Nucleic Acids; D010455:Peptides; D011073:Polyamines; C005391:diethylenetriamine; D012313:RNA; D012260:Ribonucleases