title: Fast and efficient synthesis of Zorro-LNA type 3'-5'-5'-3' oligonucleotide conjugates via parallel in situ stepwise conjugation.
abstract: Zorro-LNA is a new class of therapeutic anti-gene oligonucleotides (ONs) capable of invading supercoiled DNA. The synthesis of single stranded Zorro-LNA is typically complex and laborious, requiring reverse phosphoramidites and a chemical linker connecting the two separate ON arms. Here, a simplified synthesis strategy based on 'click chemistry' is presented with a high potential for screening Zorro-LNA ONs directed against new anti-gene targets. Four different Zorro type 3'-5' 5'-3' constructs were synthesized via parallel in situ Cu(i) [3 + 2] catalysed cycloaddition. They were prepared from commercially obtained ONs functionalized on solid support (one ON with the azide and the other ON with the activated triple bond linker N-propynoylamino)-p-toluic acid (PATA)) and after cleavage from resin, they were conjugated in solution. Our report shows the benefit of combining different approaches when developing anti-gene ONs, (1) the ability for rapid and robust screening of potential targets and (2) refining the hits with more anti-gene optimized constructs. We present as well the first report showing double-strand invasion (DSI) efficiency of two combined Zorro-LNAs.
authors.affiliation: Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, SE-141 86 Huddinge, Sweden. karin.lundin@ki.se.
authors.forename: O I
authors.initials: OI
authors.lastname: Gissberg
authors.forename: M
authors.initials: M
authors.lastname: Jezowska
authors.forename: E M
authors.initials: EM
authors.lastname: Zaghloul
authors.forename: N I
authors.initials: NI
authors.lastname: Bungsu
authors.forename: R
authors.initials: R
authors.lastname: Strömberg
authors.forename: C I E
authors.initials: CI
authors.lastname: Smith
authors.forename: K E
authors.initials: KE
authors.lastname: Lundin
authors.forename: M
authors.initials: M
authors.lastname: Honcharenko
mesh_terms: D009841:Oligonucleotides / Q000737:chemistry*
chemical_list: D009841:Oligonucleotides; C477371:locked nucleic acid