1. Academic Validation
  2. Biostable aptamers with antagonistic properties to the neuropeptide nociceptin/orphanin FQ

Biostable aptamers with antagonistic properties to the neuropeptide nociceptin/orphanin FQ

  • RNA. 2004 Mar;10(3):516-27. doi: 10.1261/rna.5186504.
Dirk Faulhammer 1 Bernd Eschgfäller Sandra Stark Petra Burgstaller Werner Englberger Jeannette Erfurth Frank Kleinjung Johanna Rupp Sebastian Dan Vulcu Werner Schröder Stefan Vonhoff Hermann Nawrath Clemens Gillen Sven Klussmann
Affiliations

Affiliation

  • 1 NOXXON Pharma AG, 10589 Berlin, Germany.
Abstract

The neuropeptide nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the opioid receptor-like 1 (ORL1) receptor, has been shown to play a prominent role in the regulation of several biological functions such as pain and stress. Here we describe the isolation and characterization of N/OFQ binding biostable RNA Aptamers (Spiegelmers) using a mirror-image in vitro selection approach. Spiegelmers are L-enantiomeric oligonucleotide ligands that display high affinity and specificity to their targets and high resistance to enzymatic degradation compared to D-oligonucleotides. A representative Spiegelmer from the selections performed was size-minimized to two distinct sequences capable of high affinity binding to N/OFQ. The Spiegelmers were shown to antagonize binding of N/OFQ to the ORL1 receptor in a binding-competition assay. The calculated IC(50) values for the Spiegelmers NOX 2149 and NOX 2137a/b were 110 nM and 330 nM, respectively. The competitive antagonistic properties of these Spiegelmers were further demonstrated by their effective and specific inhibition of G-protein activation in two additional models. The Spiegelmers antagonized the N/OFQ-induced GTPgammaS incorporation into cell membranes of a CHO-K1 cell line expressing the human ORL1 receptor. In oocytes from Xenopus laevis, NOX 2149 showed an antagonistic effect to the N/OFQ-ORL 1 receptor system that was functionally coupled with G-protein-regulated inwardly rectifying K(+) channels.

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