title: Selective amplification of glucocorticoid anti-inflammatory activity through synergistic multi-target action of a combination drug.
abstract: BACKGROUND
  Glucocorticoids are a mainstay of anti-inflammatory therapy, but significant adverse effects ultimately limit their utility. Previous efforts to design glucocorticoid structures with an increased therapeutic window have focused on dissociating anti-inflammatory transcriptional repression from adverse effects primarily driven by transcriptional activation. An alternative to this medicinal chemistry approach is a systems biology based strategy that seeks to amplify selectively the anti-inflammatory activity of very low dose glucocorticoid in immune cells without modulating alternative cellular networks that mediate glucocorticoid toxicity.
  
  
  METHODS
  The combination of prednisolone and the antithrombotic drug dipyridamole was profiled using in vitro and in vivo models of anti-inflammatory activity and glucocorticoid-induced adverse effects to demonstrate a dissociated activity profile.
  
  
  RESULTS
  The combination synergistically suppresses release of proinflammatory mediators, including tumour necrosis factor-alpha, IL-6, chemokine (C-C motif) ligand 5 (RANTES), matrix metalloproteinase-9, and others, from human peripheral blood mononuclear cells and mouse macrophages. In rat models of acute lipopolysaccharide-induced endotoxemia and delayed-type hypersensitivity, and in chronic models of collagen-induced and adjuvant-induced arthritis, the combination produced anti-inflammatory activity that required only a subtherapeutic dose of prednisolone. The immune-specific amplification of prednisolone anti-inflammatory activity by dipyridamole did not extend to glucocorticoid-mediated adverse effects, including corticosterone suppression or increased expression of tyrosine aminotransferase, in vivo after repeat dosing in rats. After 8 weeks of oral dosing in mice, treatment with the combination did not alter prednisolone-induced reduction in osteocalcin and mid-femur bone density, which are markers of steroid-induced osteoporosis. Additionally, amplification was not observed in the cellular network of corticotroph AtT-20/D16v-F2 cells in vitro, as measured by pro-opiomelanocortin expression and adrenocorticotropic hormone secretion.
  
  
  CONCLUSIONS
  These data suggest that the multi-target mechanism of low-dose prednisolone and dipyridamole creates a dissociated activity profile with an increased therapeutic window through cellular network selective amplification of glucocorticoid-mediated anti-inflammatory signaling.
authors.affiliation: CombinatoRx, Incorporated, First Street, Cambridge, MA 02142, USA. gzimmermann@combinatorx.com
authors.forename: Grant R
authors.initials: GR
authors.lastname: Zimmermann
authors.forename: William
authors.initials: W
authors.lastname: Avery
authors.forename: Alyce L
authors.initials: AL
authors.lastname: Finelli
authors.forename: Melissa
authors.initials: M
authors.lastname: Farwell
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Fraser
authors.forename: Alexis A
authors.initials: AA
authors.lastname: Borisy
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D004176:Dipyridamole / Q000494:pharmacology*; D004359:Drug Therapy, Combination; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D007249:Inflammation / Q000188:drug therapy*; D018836:Inflammation Mediators / Q000276:immunology; D051379:Mice; D011239:Prednisolone / Q000494:pharmacology*; D051381:Rats; D011917:Rats, Inbred Lew
chemical_list: D000893:Anti-Inflammatory Agents; D005938:Glucocorticoids; D018836:Inflammation Mediators; D004176:Dipyridamole; D011239:Prednisolone