title: Antisense oligonucleotides containing conformationally constrained 2',4'-(N-methoxy)aminomethylene and 2',4'-aminooxymethylene and 2'-O,4'-C-aminomethylene bridged nucleoside analogues show improved potency in animal models.
abstract: To identify chemistries and strategies to improve the potency of MOE second generation ASOs, we have evaluated gapmer antisense oligonucleotides containing BNAs having N-O bonds. These modifications include N-MeO-amino BNA, N-Me-aminooxy BNA, 2',4'-BNA(NC)[NMe], and 2',4'-BNA(NC) bridged nucleoside analogues. These modifications provided increased thermal stability and improved in vitro activity compared to the corresponding ASO containing the MOE modification. Additionally, ASOs containing N-MeO-amino BNA, N-Me-aminooxy BNA, and 2',4'-BNA(NC)[NMe] modifications showed improved in vivo activity (>5-fold) compared to MOE ASO. Importantly, toxicity parameters, such as AST, ALT, liver, kidney, and body weights, were found to be normal for N-MeO-amino BNA, N-Me-aminooxy BNA, and 2',4'-BNA(NC)[NMe] ASO treated animals. The data generated in these experiments suggest that N-MeO-amino BNA, N-Me-aminooxy BNA, and 2',4'-BNA(NC)[NMe] are useful modifications for applications in both antisense and other oligonucleotide based drug discovery efforts.
authors.affiliation: Department of Medicinal Chemistry and Antisense Core Research, Isis Pharmaceuticals Inc., 1896 Rutherford Road, Carlsbad, California 92008, USA. tprakash@isisph.com
authors.forename: Thazha P
authors.initials: TP
authors.lastname: Prakash
authors.forename: Andrew
authors.initials: A
authors.lastname: Siwkowski
authors.forename: Charles R
authors.initials: CR
authors.lastname: Allerson
authors.forename: Michael T
authors.initials: MT
authors.lastname: Migawa
authors.forename: Sam
authors.initials: S
authors.lastname: Lee
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Gaus
authors.forename: Chris
authors.initials: C
authors.lastname: Black
authors.forename: Punit P
authors.initials: PP
authors.lastname: Seth
authors.forename: Eric E
authors.initials: EE
authors.lastname: Swayze
authors.forename: Balkrishen
authors.initials: B
authors.lastname: Bhat
mesh_terms: D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D001835:Body Weight / Q000187:drug effects; D002460:Cell Line; D004355:Drug Stability; D006358:Hot Temperature; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008968:Molecular Conformation; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry; D020320:Oligoribonucleotides, Antisense / Q000138:chemical synthesis* / Q000494:pharmacology / Q000633:toxicity; D009929:Organ Size / Q000187:drug effects; D051059:PTEN Phosphohydrolase / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D013329:Structure-Activity Relationship
chemical_list: D009692:Nucleic Acid Heteroduplexes; D020320:Oligoribonucleotides, Antisense; D012333:RNA, Messenger; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D051059:PTEN Phosphohydrolase