title: Mapracorat, a novel selective glucocorticoid receptor agonist, inhibits hyperosmolar-induced cytokine release and MAPK pathways in human corneal epithelial cells.
abstract: OBJECTIVE
  Increasing evidence suggests that tear hyperosmolarity is a central mechanism causing ocular surface inflammation and damage in dry eye disease. Mapracorat (BOL-303242-X) is a novel glucocorticoid receptor agonist currently under clinical evaluation for use in the treatment of dry eye disease. This study assessed the anti-inflammatory effects of mapracorat in an in vitro osmotic stress model which mimics some of the pathophysiological changes seen in dry eye.
  
  
  METHODS
  Human corneal epithelial cells were cultured in normal osmolar media (317 mOsM) or 440 mOsM hyperosmolar media for 24 h. Luminex technology was used to determine the effect of mapracorat on hyperosmolar-induced cytokine release. Effects of mapracorat on mitogen-activated protein kinase (MAPK) phosphorylation were determined by cell based ELISA. Effects of mapracorat on nuclear factor kappa B (NFkappaB) and activator protein-1 (AP-1) transcriptional activity were assessed by reporter gene assay. Dexamethasone was used as a control.
  
  
  RESULTS
  Hyperosmolar conditions induced release of the pro-inflammatory cytokines interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) from cultured human corneal epithelial cells, and altered the phosphorylation state of p38 and c-Jun N-terminal kinase (JNK) and transcriptional activity of NFkappaB and AP-1. Incubation of cells with mapracorat inhibited hyperosmolar-induced cytokine release with comparable activity and potency as dexamethasone. This inhibition was reversed by the glucocorticoid receptor antagonist mifepristone (RU-486). Increased phosphorylation of p38 and JNK caused by hyperosmolarity was inhibited by mapracorat. Mapracorat also significantly decreased the hyperosmolar-induced increase in NFkappaB and AP-1 transcriptional activity.
  
  
  CONCLUSIONS
  Mapracorat acts as a potent anti-inflammatory agent in corneal epithelial cells challenged with osmotic stress, with comparable activity to the traditional steroid dexamethasone. These in vitro data suggest that mapracorat may be efficacious in the treatment of dry eye disease.
authors.affiliation: Pharmaceutical R&D, Bausch & Lomb, Inc., Rochester, NY 14609, USA.
authors.forename: Megan E
authors.initials: ME
authors.lastname: Cavet
authors.forename: Karen L
authors.initials: KL
authors.lastname: Harrington
authors.forename: Keith W
authors.initials: KW
authors.lastname: Ward
authors.forename: Jin-Zhong
authors.initials: JZ
authors.lastname: Zhang
mesh_terms: D001572:Benzofurans / Q000494:pharmacology*; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology*; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D015735:Mifepristone / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D009994:Osmolar Concentration; D000439:Pentanols / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011804:Quinolines / Q000494:pharmacology*; D011965:Receptors, Glucocorticoid / Q000819:agonists*; D018808:Transcription Factor AP-1 / Q000378:metabolism
chemical_list: D001572:Benzofurans; D016207:Cytokines; 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; D018808:Transcription Factor AP-1; D015735:Mifepristone; D003907:Dexamethasone; D020928:Mitogen-Activated Protein Kinases
references.citation: Optometry. 2009 Oct;80(10):555-66
references.pmid: 19801339
references.citation: Int Ophthalmol Clin. 1994 Winter;34(1):27-36
references.pmid: 8169071
references.citation: Invest Ophthalmol Vis Sci. 2009 Aug;50(8):3671-9
references.pmid: 19324847
references.citation: Br J Pharmacol. 2009 Oct;158(4):1088-103
references.pmid: 19422381
references.citation: Invest Ophthalmol Vis Sci. 2009 Aug;50(8):3802-7
references.pmid: 19339740
references.citation: Eye Contact Lens. 2005 Sep;31(5):186-93
references.pmid: 16163009
references.citation: J Steroid Biochem Mol Biol. 2010 May 31;120(2-3):96-104
references.pmid: 20206690
references.citation: Am J Respir Cell Mol Biol. 2008 Aug;39(2):208-17
references.pmid: 18314542
references.citation: IUBMB Life. 2005 Apr-May;57(4-5):283-95
references.pmid: 16036612
references.citation: J Biopharm Stat. 1998 Nov;8(4):645-65
references.pmid: 9855040
references.citation: Curr Eye Res. 2010 Feb;35(2):165-75
references.pmid: 20136427
references.citation: Exp Eye Res. 2010 Mar;90(3):437-43
references.pmid: 20026325
references.citation: Mol Cell Endocrinol. 2007 Sep 15;275(1-2):71-8
references.pmid: 17630118
references.citation: Endocr Rev. 2003 Aug;24(4):488-522
references.pmid: 12920152
references.citation: J Immunol Methods. 2000 Sep 21;243(1-2):243-55
references.pmid: 10986418
references.citation: J Immunol Methods. 1994 Apr 15;170(2):211-24
references.pmid: 8157999
references.citation: Cornea. 2008 Jun;27(5):574-9
references.pmid: 18520508
references.citation: Mol Vis. 2009 Dec 08;15:2606-16
references.pmid: 20011631
references.citation: Invest Ophthalmol Vis Sci. 2004 Dec;45(12):4293-301
references.pmid: 15557435
references.citation: J Leukoc Biol. 2002 Jan;71(1):9-15
references.pmid: 11781376
references.citation: Cornea. 2007 May;26(4):431-7
references.pmid: 17457192
references.citation: FEMS Immunol Med Microbiol. 2007 Oct;51(1):171-84
references.pmid: 17645739
references.citation: Invest Ophthalmol Vis Sci. 2010 Jun;51(6):3083-91
references.pmid: 20130281
references.citation: Ocul Surf. 2009 Apr;7(2):78-92
references.pmid: 19383277
references.citation: Trends Endocrinol Metab. 2006 Oct;17(8):321-7
references.pmid: 16942889
references.citation: Curr Top Med Chem. 2008;8(9):813-38
references.pmid: 18537690
references.citation: J Exp Med. 2006 Aug 7;203(8):1883-9
references.pmid: 16880258
references.citation: Curr Opin Cell Biol. 1997 Apr;9(2):180-6
references.pmid: 9069255
references.citation: Am J Ophthalmol. 2009 Feb;147(2):198-205. e1
references.pmid: 18992869
references.citation: Biotechnol Annu Rev. 2006;12:269-300
references.pmid: 17045197
references.citation: Arch Ophthalmol. 2009 Jul;127(7):882-7
references.pmid: 19597109
references.citation: Ocul Surf. 2007 Apr;5(2):75-92
references.pmid: 17508116
references.citation: Exp Eye Res. 2006 Apr;82(4):588-96
references.pmid: 16202406
references.citation: Cornea. 2009 Oct;28(9):1023-7
references.pmid: 19724208
references.citation: Eur J Pharmacol. 2008 Apr 7;583(2-3):290-302
references.pmid: 18289525
references.citation: Cornea. 2007 Jun;26(5):579-84
references.pmid: 17525655
references.citation: Invest Ophthalmol Vis Sci. 2004 Dec;45(12):4302-11
references.pmid: 15557436
references.citation: Eur J Biochem. 2000 Sep;267(17):5421-6
references.pmid: 10951200
references.citation: J Mol Endocrinol. 2008 Oct;41(4):239-49
references.pmid: 18682532
references.citation: Mol Cell Endocrinol. 2007 Sep 15;275(1-2):109-17
references.pmid: 17630119
references.citation: Invest Ophthalmol Vis Sci. 2002 Aug;43(8):2609-14
references.pmid: 12147592
references.citation: Invest Ophthalmol Vis Sci. 2010 Jan;51(1):437-46
references.pmid: 19696178
references.citation: Clin Immunol. 2004 Mar;110(3):252-66
references.pmid: 15047203
references.citation: Clin Ophthalmol. 2008 Dec;2(4):829-36
references.pmid: 19668437
references.citation: Invest Ophthalmol Vis Sci. 2010 Feb;51(2):643-50
references.pmid: 19850844
references.citation: Steroids. 2008 Oct;73(9-10):1025-9
references.pmid: 18221974