title: Novel cationic lipophilic peptides for oligodeoxynucleotide delivery.
abstract: In search of new oligodeoxynucleotide (ODN) delivery agents, we evaluated novel peptides derived from core peptide H-GLRILLLKV-OH (CP). CP is a fragment designed from the T-cell antigen receptor (TCR) alpha-chain transmembrane sequence. CP was able to enter cells including T-cells and inhibited interleukin-2 (IL-2) production. To examine the effect of increased lipophilicity on cellular uptake and activity of CP, a lipoamino acid (2-aminododecanoic acid) was incorporated into peptide CP resulting in 2-aminodecanoyl-CP (LP). The toxicity of CP and LP was assessed by measuring the haemolytic activity. Neither compound caused any haemolysis of red blood cells. We have also compared the biological activities of the CP and LP. Using a T-cell antigen presentation assay, the more lipophilic LP caused greater inhibition of IL-2 production than the parent CP in the antigen stimulated T-cells. The LP also showed increased permeability than CP in the Caco-2 cell assay. We utilised the enhanced cell permeability property of LP in oligodeoxynucleotide ODN1 delivery. Isothermal titration calorimetry (ITC) suggested that CP and LP complex with ODN1 in a 12:1 (CP:ODN1) and 15:1 (LP:ODN1) ratio. These complexes were then transfected into human retinal pigment epithelial cells. The level of transfection was measured by the decreased production of the protein human vascular endothelial growth factor (hVEGF). The results revealed greater transfection efficiency for both CP and LP (47%, 55% more inhibition) compared to commercially available transfection agent cytofectin GSV. These results suggested that the CP and particularly its lipophilic analogue LP have the potential to be used as oligodeoxynucleotide delivery systems.
authors.affiliation: School of Pharmacy, Griffith University, Gold Coast Campus, PMB 50 Gold Coast Mail Centre, QLD 9726, Australia.
authors.forename: Enoch
authors.initials: E
authors.lastname: Chan
authors.forename: Michael
authors.initials: M
authors.lastname: Amon
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Marano
authors.forename: Norbert
authors.initials: N
authors.lastname: Wimmer
authors.forename: Philip S
authors.initials: PS
authors.lastname: Kearns
authors.forename: Nicholas
authors.initials: N
authors.lastname: Manolios
authors.forename: P Elizabeth
authors.initials: PE
authors.lastname: Rakoczy
authors.forename: Istvan
authors.initials: I
authors.lastname: Toth
mesh_terms: D001483:Base Sequence; D018938:Caco-2 Cells; D002151:Calorimetry; D002412:Cations; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis; D009841:Oligonucleotides / Q000008:administration & dosage*; D010455:Peptides / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism
chemical_list: D002412:Cations; D007376:Interleukin-2; D009841:Oligonucleotides; D010455:Peptides