The solution synthesis of antisense oligonucleotide-peptide conjugates directly linked via phosphoramide bond by using a fragment coupling approach.
To improve antisense oligonucleotide penetration inside cells, conjugates of oligonucleotides and cell-penetrating peptides, covalently linked through a phosphoramide bond, were prepared by a fragment coupling approach in the liquid phase. Two methods were used for this synthesis, i.e., phosphorylation of a peptide amino group by an oligonucleotide terminal phosphate 1-hydroxybenzotriazole ester in aqueous media or condensation of phosphate and amino groups in presence of triphenylphosphine, 2,2'-dithiopyridine and 4-dimethylaminopyridine in organic media. Several oligonucleotides, including a 18-mer antisense oligodeoxyribonucleotide complementary to an internal coding region of the reporter gene of the green fluorescent protein (GFP) were prepared. Peptides derived from the third helix of the homeodomain of Antennapedia, the influenza envelope hemagglutinin subunit as well as melittin and polymyxin B were used for the conjugates' synthesis. The peptides with various amino acid composition were chosen to confirm that these coupling methods are of a general use.
Sumbatyan
Nataliya V
NV
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russian Federation. sumbtyan@belozersky.msu.ru
Mandrugin
Vassili A
VA
Deroussent
Alain
A
Bertrand
Jean-Rémi
JR
Majer
Zsuzsa
Z
Malvy
Claude
C
Korshunova
Galina A
GA
Hollosi
Miklos
M
Gottikh
Marina B
MB
D000577:Amides / Q000737:chemistry*; D015394:Molecular Structure; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis* / Q000737:chemistry; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry; D063088:Phosphoramides; D010756:Phosphoric Acids / Q000737:chemistry*; D010766:Phosphorylation; D012996:Solutions / Q000138:chemical synthesis
D000577:Amides; D016376:Oligonucleotides, Antisense; D010446:Peptide Fragments; D063088:Phosphoramides; D010756:Phosphoric Acids; D012996:Solutions; C005438:phosphoramide