Synthesis of high quality phosphorothioate oligonucleotides as antisense drugs. Use of I-linker in the elimination of 3'-terminal phosphorothioate monoesters.
Detritylation of a 5'-O-DMT-2'-deoxyadenosine moiety attached to solid support under acidic condition leads to depurination during oligonucleotide synthesis. Deprotection followed by reversed phase HPLC purification leads to desired oligonucleotide contaminated with significant levels of 3'-terminal phosphorothiaote (3'-TPT) monoester (n-1)-mer. However, it is demonstrated that attachment of dA nucleoside through its exocyclic amino group to solid support leads to substantial reduction of 3'-TPT formation thereby improving the quality of oligonucleotide synthesized.
Ravikumar
Vasulinga T
VT
Isis Pharmaceuticals, Carlsbad, California 92009, USA. vravikumar@isisph.com
Kumar
R Krishna
RK
Capaldi
Daniel C
DC
Cole
Douglas L
DL
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D007202:Indicators and Reagents; D008968:Molecular Conformation; D015394:Molecular Structure; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis* / Q000737:chemistry; D010710:Phosphates
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D010710:Phosphates; C027903:oligo (dT)