title: 2'-Deoxy purine, 2'-O-methyl pyrimidine (dRmY) aptamers as candidate therapeutics.
abstract: Aptamers are short oligonucleotides that fold into well-defined three-dimensional architectures thereby enabling specific binding to molecular targets such as proteins. To be successful as a novel therapeutic modality, it is important for aptamers to not only bind their targets with high specificity and affinity, but also to exhibit favorable properties with respect to in vivo stability, cost-effective synthesis, and tolerability (i.e., safety). We describe methods for generating aptamers comprising 2 - deoxy purines and 2 -O-methyl pyrimidines (dRmY) that broadly satisfy many of these additional constraints. Conditions under which dRmY transcripts can be efficiently synthesized using mutant T7 RNA polymerases have been identified and used to generate large libraries from which dRmY aptamers to multiple target proteins, including interleukin (IL)-23 and thrombin, have been successfully discovered using the SELEX process. dRmY aptamers are shown to be highly nuclease-resistant, long-lived in vivo, efficiently synthesized, and capable of binding protein targets in a manner that inhibits their biologic activity with K(D) values in the low nM range. We believe that dRmY aptamers have considerable potential as a new class of therapeutic aptamers.
authors.affiliation: Archemix Corporation, Cambridge, MA 02142, USA.
authors.forename: Paula E
authors.initials: PE
authors.lastname: Burmeister
authors.forename: Chunhua
authors.initials: C
authors.lastname: Wang
authors.forename: Jason R
authors.initials: JR
authors.lastname: Killough
authors.forename: Scott D
authors.initials: SD
authors.lastname: Lewis
authors.forename: Lillian R
authors.initials: LR
authors.lastname: Horwitz
authors.forename: Alicia
authors.initials: A
authors.lastname: Ferguson
authors.forename: Kristin M
authors.initials: KM
authors.lastname: Thompson
authors.forename: P Shannon
authors.initials: PS
authors.lastname: Pendergrast
authors.forename: Thomas G
authors.initials: TG
authors.lastname: McCauley
authors.forename: Markus
authors.initials: M
authors.lastname: Kurz
authors.forename: John
authors.initials: J
authors.lastname: Diener
authors.forename: Sharon T
authors.initials: ST
authors.lastname: Cload
authors.forename: Charles
authors.initials: C
authors.lastname: Wilson
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Keefe
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use*; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D004355:Drug Stability; D006801:Humans; D051379:Mice; D015394:Molecular Structure; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D052156:SELEX Aptamer Technique; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases