title: A novel CGRP-neutralizing Spiegelmer attenuates neurogenic plasma protein extravasation.
abstract: OBJECTIVE
  Calcitonin gene-related peptide (CGRP) plays an important role in the pathology of migraine, and recent clinical trials suggest the inhibition of CGRP-mediated processes as a new therapeutic option in migraine. In this study, we describe the generation of NOX-L41, a CGRP-neutralizing mirror-image (L-)aptamer (Spiegelmer) and investigate its in vitro and in vivo function.
  
  
  METHODS
  A CGRP-binding Spiegelmer was identified by in vitro selection. Binding studies were performed using surface plasmon resonance (SPR), and the inhibitory activity was determined in cell-based assays. The pharmacokinetic profile comparing i.v. and s.c. dosing was analysed in rats. Intravital two-photon microscopy was employed to follow extravasation from meningeal vessels. Finally, in vivo efficacy was tested in a model of electrically evoked meningeal plasma protein extravasation (PPE) in rats.
  
  
  RESULTS
  We identified NOX-L41, a novel CGRP-neutralizing Spiegelmer. SPR studies showed that NOX-L41 binds to human and rat/mouse CGRP with sub-nanomolar affinities and is highly selective against related peptides such as amylin. In vitro, NOX-L41 effectively inhibited CGRP-induced cAMP formation in SK-N-MC cells. In rats, NOX-L41 had a plasma half-life of 8 h. Pharmacodynamic studies showed that NOX-L41 extravasates from blood vessels in the dura mater and inhibits neurogenic meningeal PPE for at least 18 h after single dosing.
  
  
  CONCLUSIONS
  This is the first description of the CGRP-neutralizing Spiegelmer NOX-L41. Preclinical studies confirmed a role for CGRP in neurogenic PPE and provided proof-of-concept for the potential use of this new drug candidate for the treatment or prevention of migraine.
authors: NOXXON Pharma AG, Berlin, Germany.
authors: K
authors: http://orcid.org/0000-0002-4638-4826
authors: K
authors: Hoehlig
authors: Eli Lilly and Company, Indianapolis, IN, USA.
authors: K W
authors: KW
authors: Johnson
authors: Neurotar Ltd, Helsinki, Finland.
authors: E
authors: E
authors: Pryazhnikov
authors: NOXXON Pharma AG, Berlin, Germany.
authors: C
authors: C
authors: Maasch
authors: Eli Lilly and Company, Indianapolis, IN, USA.
authors: A
authors: A
authors: Clemens-Smith
authors: NOXXON Pharma AG, Berlin, Germany.
authors: W G
authors: WG
authors: Purschke
authors: NOXXON Pharma AG, Berlin, Germany.
authors: S
authors: S
authors: Vauléon
authors: NOXXON Pharma AG, Berlin, Germany.
authors: K
authors: K
authors: Buchner
authors: NOXXON Pharma AG, Berlin, Germany.
authors: F
authors: F
authors: Jarosch
authors: Neurotar Ltd, Helsinki, Finland.
authors: L
authors: L
authors: Khiroug
authors: NOXXON Pharma AG, Berlin, Germany.
authors: A
authors: A
authors: Vater
authors: NOXXON Pharma AG, Berlin, Germany.
authors: S
authors: S
authors: Klussmann
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D001798:Blood Proteins / Q000378:metabolism*; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D058228:Islet Amyloid Polypeptide / Q000378:metabolism; D008297:Male; D008578:Meninges / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D020349:Surface Plasmon Resonance; D013997:Time Factors
chemical_list: D052157:Aptamers, Nucleotide; D001798:Blood Proteins; D058228:Islet Amyloid Polypeptide; C000605332:NOX-L41; D000242:Cyclic AMP; D015740:Calcitonin Gene-Related Peptide