title: BOL-303242-X, a novel selective glucocorticoid receptor agonist, with full anti-inflammatory properties in human ocular cells.
abstract: OBJECTIVE
  BOL-303242-X is a novel selective glucocorticoid receptor agonist under clinical evaluation for the treatment of inflammatory skin and eye diseases. Data from in vitro and in vivo studies suggest an improved side-effect profile of this compound compared to classical glucocorticoids. The aim of this study was to determine the anti-inflammatory effect of BOL-303242-X in ocular cells.
  
  
  METHODS
  Four primary human ocular cell cultures, including human conjunctival fibroblasts (HConFs), human corneal epithelial cells (HCEpiCs), human optic nerve astrocytes (HONAs), and human retinal endothelial cells (HRECs), as well as a human monocytic cell line, THP-1, were challenged with either lipopolysacharide (LPS) or interleukin-1ss (IL-1ss). Luminex technology was used to determine the effect of BOL-303242-X on LPS- or IL-1ss-induced cytokine release and intercellular adhesion molecule-1 (ICAM-1) levels. Effects of BOL-303242-X on nuclear factor kappa B (NFkappaB) and mitogen-activated protein kinase (MAPK) in HCEpiCs were also assessed by measuring inhibitory kappa B protein-alpha (IkappaB-alpha), phosphorylated p65 NFkappaB, and MAPK levels by western blotting. Dexamethasone (DEX) or triamcinolone acetonide (TA) was used as the control.
  
  
  RESULTS
  LPS or IL-1ss induced multiple cytokine release in all cell types studied. BOL-303242-X significantly reduced LPS- or IL-1ss-induced inflammatory cytokine release in a dose-dependent manner, including granulocyte colony-stimulating factor (G-CSF), IL-1ss, IL-6, IL-8, IL-12p40, monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-alpha). BOL-303242-X showed activity and potency comparable to that observed for DEX or TA. A statistically significant inhibitory effect of BOL-303242-X was observed at doses ranging from 1 to 100 nM in HConFs, HCEpiCs, HONAs, and THP-1. The IC(50) values for these effects were in the low nM range. BOL-303242-X also significantly reduced LPS-induced IL-1ss release and ICAM-1 levels in HRECs. Furthermore, BOL-303242-X inhibited IL-1ss-induced decreases in IkappaB-alpha levels, as well as IL-1ss-induced phosphorylation of NFkappaB, p38, and c-Jun-N-terminal kinase (JNK) MAPKs in HCEpiCs.
  
  
  CONCLUSIONS
  BOL-303242-X acts as a potent anti-inflammatory agent in various primary human ocular cells with similar activity and potency compared to classical steroids. Results also suggest that MAPK (p38 and JNK) and NFkappaB signaling pathways are involved in the anti-inflammatory properties of BOL-303242-X in HCEpiCs. An improved side effect profile of this novel SEGRA compound has been reported recently. Thus, BOL-303242-X may provide a new option for the treatment of ophthalmic conditions with an inflammatory component.
authors.affiliation: Pharmaceutical R&D, Bausch & Lomb Inc, Rochester, NY 14603, USA. jinzhong_zhang@bausch.com
authors.forename: Jin-Zhong
authors.initials: JZ
authors.lastname: Zhang
authors.forename: Megan E
authors.initials: ME
authors.lastname: Cavet
authors.forename: Karl R
authors.initials: KR
authors.lastname: VanderMeid
authors.forename: Mercedes
authors.initials: M
authors.lastname: Salvador-Silva
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: López
authors.forename: Keith W
authors.initials: KW
authors.lastname: Ward
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D001572:Benzofurans / Q000494:pharmacology*; D002457:Cell Extracts; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology; D003907:Dexamethasone / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D019573:Epithelium, Corneal / Q000166:cytology; D005123:Eye / Q000166:cytology* / Q000187:drug effects*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009900:Optic Nerve / Q000166:cytology; D000439:Pentanols / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011804:Quinolines / Q000494:pharmacology*; D011965:Receptors, Glucocorticoid / Q000819:agonists*; D014222:Triamcinolone Acetonide / Q000494:pharmacology
chemical_list: D000893:Anti-Inflammatory Agents; D001572:Benzofurans; D002457:Cell Extracts; D020672:I-kappa B Proteins; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D000439:Pentanols; D011804:Quinolines; C546413:R-1,1,1-trifluoro-4-(5-fluoro-2,3-dihydrobenzofuran-7-yl)-4-methyl-2-(((2-methyl-5-quinolyl)amino)methyl)pentan-2-ol; D011965:Receptors, Glucocorticoid; D018799:Intercellular Adhesion Molecule-1; D003907:Dexamethasone; D020928:Mitogen-Activated Protein Kinases; D014222:Triamcinolone Acetonide