2'-Deoxy purine, 2'-O-methyl pyrimidine (dRmY) aptamers as candidate therapeutics.
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.
Burmeister
Paula E
PE
Archemix Corporation, Cambridge, MA 02142, USA.
Wang
Chunhua
C
Killough
Jason R
JR
Lewis
Scott D
SD
Horwitz
Lillian R
LR
Ferguson
Alicia
A
Thompson
Kristin M
KM
Pendergrast
P Shannon
PS
McCauley
Thomas G
TG
Kurz
Markus
M
Diener
John
J
Cload
Sharon T
ST
Wilson
Charles
C
Keefe
Anthony D
AD
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
D052157:Aptamers, Nucleotide; D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases