Chemogenomic identification of Ref-1/AP-1 as a therapeutic target for asthma.
Asthma is characterized by an oxidantantioxidant imbalance in the lungs leading to activation of redox-sensitive transcription factors, nuclear factor kappaB (NF-kappaB), and activator protein-1 (AP-1). To develop therapeutic strategies for asthma, we used a chemogenomics approach to screen for small molecule inhibitor(s) of AP-1 transcription. We developed a beta-strand mimetic template that acts as a reversible inhibitor (pseudosubstrate) of redox proteins. This template incorporates an enedione moiety to trap reactive cysteine nucleophiles in the active sites of redox proteins. Specificity for individual redox factors was achieved through variations in X and Y functionality by using a combinatorial library approach. A limited array (2 x 6) was constructed where X was either NHCH(3) or NHCH(2) Ph and Y was methyl, phenyl, m-cyanophenyl, m-nitrophenyl, m-acetylaniline, or m-methylbenzoate. These analogs were evaluated for their ability to inhibit transcription in transiently transfected human lung epithelial A549 cells from either an AP-1 or NF-kappaB reporter. A small-molecule inhibitor, PNRI-299, was identified that selectively inhibited AP-1 transcription (IC(50) of 20 microM) without affecting NF-kappaB transcription (up to 200 microM) or thioredoxin (up to 200 microM). The molecular target of PNRI-299 was determined to be the oxidoreductase, redox effector factor-1 by an affinity chromatography approach. The selective redox effector factor-1 inhibitor, PNRI-299, significantly reduced airway eosinophil infiltration, mucus hypersecretion, edema, and IL-4 levels in a mouse asthma model. These data validate AP-1 as an important therapeutic target in allergic airway inflammation.
Nguyen
Cu
C
Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA.
Teo
Jia-Ling
JL
Matsuda
Akihisa
A
Eguchi
Masakatsu
M
Chi
Emil Y
EY
Henderson
William R
WR
Kahn
Michael
M
D000818:Animals; D001249:Asthma / Q000188:drug therapy*; D001665:Binding Sites; D001992:Bronchoalveolar Lavage Fluid; D019757:Carbon-Oxygen Lyases / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002845:Chromatography; D003545:Cysteine / Q000737:chemistry; D003600:Cytosol / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D005260:Female; D017930:Genes, Reporter; D006801:Humans; D020128:Inhibitory Concentration 50; D015847:Interleukin-4 / Q000378:metabolism; D008168:Lung / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008956:Models, Chemical; D008958:Models, Molecular; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D010084:Oxidation-Reduction; D019151:Peptide Library; D013879:Thioredoxins / Q000037:antagonists & inhibitors; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016328:NF-kappa B; D019151:Peptide Library; D018808:Transcription Factor AP-1; D015847:Interleukin-4; D013879:Thioredoxins; D019757:Carbon-Oxygen Lyases; C085108:APEX1 protein, human; C486216:Apex1 protein, mouse; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D003545:Cysteine