title: Enabling Multiple Conjugation to Oligonucleotides Using "Click Cycles".
abstract: An efficient method for the synthesis of multiply functionalized oligonucleotides (ONs) utilizing a novel H-phosphonate alkyne-based linker for multiple functionalization (LMF) is developed. The strategy allows for the conjugation of various active entities to oligonucleotide through the postsynthetic attachment of LMF at the 5'-terminus of ONs using H-phosphonate chemistry followed by conjugation of various entities via [3 + 2] copper(I) catalyzed cycloaddition in a stepwise manner. Each cycle is composed of attachment of the LMF followed by a click reaction with azido-containing units. Sequential solid-phase synthesis of oligonucleotide conjugates containing three attached entities was performed using an acetylated form of MIF peptide conjugated to azido linker, achieving high conversions at each unit addition. In addition, to show the versatility of the method, oligonucleotide conjugates with several different classes of compounds were synthesized. Each conjugate containing three different entities, whose structure and function varied (e.g., sugars, peptides, fluorescent labels, and m3G-Caps).
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum , SE-14183 Huddinge, Sweden.
authors.forename: Martina
authors.initials: M
authors.lastname: Jezowska
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum , SE-14183 Huddinge, Sweden.
authors.forename: Dmytro
authors.initials: D
authors.lastname: Honcharenko
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum , SE-14183 Huddinge, Sweden.
authors.forename: Alice
authors.initials: A
authors.lastname: Ghidini
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum , SE-14183 Huddinge, Sweden.
authors.forename: Roger
authors.initials: R
authors.lastname: Strömberg
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum , SE-14183 Huddinge, Sweden.
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Honcharenko
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002384:Catalysis; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry*; D010757:Phosphorous Acids / Q000737:chemistry
chemical_list: D000480:Alkynes; D001386:Azides; D009841:Oligonucleotides; D010757:Phosphorous Acids; C570063:phosphonic acid; D003300:Copper