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Discovery of novel inhibitors of the phosphatase activity of the soluble epoxide hydrolase
Author(s) -
Kramer Jan Sebastian,
Woltersdorf Stefano,
Wittmann Sandra,
Hiesinger Kerstin,
Klingler Franca,
Heering Jan,
Merk Daniel,
Chaikuad Apirat,
Pogoryelov Denys,
Steinhilber Dieter,
Rovati G. Enrico,
Knapp Stefan,
Proschak Ewgenij
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.558.3
Subject(s) - epoxide hydrolase 2 , hydrolase , phosphatase , chemistry , epoxide hydrolase , enzyme , dual specificity phosphatase , linker , alkaline phosphatase , biochemistry , docking (animal) , microsome , medicine , nursing , computer science , operating system
The soluble epoxide Hydrolase (sEH) is an emerging pharmacological target. The enzyme is an antiparallel homodimer, where each of the two subunits is composed of a C‐terminal hydrolase domain connected to an N‐terminal phosphatase domain by a proline‐rich linker. (Cronin et al. 2003) Although the physiological role of the hydrolase domain is well‐investigated (Harris und Hammock 2013, S. 61), little is known about the phosphatase activity. One of the main reasons for this is the lack of high‐affinity inhibitors, which can be used as chemical probes (Arrowsmith et al. 2015) to investigate its physiological role. In this paper, we present a new series of inhibitors for the human sEH‐phosphatase domain. Starting from an HTS hit Oxaprozin we expanded the SAR using a fluorescence‐based activity assay (Klingler et al. 2016). For one of the most potent inhbitors SWE101, we made ITC measurements to further characterize the interactions of the compound with the phosphatase. The compound binds with a Kd of 0.3 μM with balanced enthalpic and entropic contributions. In differential scanning fluorimetry (DSF) experiments a strong stabilizing effect of the two most potent inhibitors SWE101 and SWE102 on the phosphatase domain was visible, while there was only a slight effect on the stability of the hydrolase domain. These two inhibitors have been further tested on a number of potential lipid signaling targets including PPARα, PPARδ, PPARγ, FXR, RXRα, LXRα, LXRβ, COX 1, and COX 2. Additionally, we were able to co‐crystalize the phosphatase domain with one of our most potent inhibitors SWE102 under acidic conditions and without phosphate in the buffer. To our best knowledge, this study is the first description of the interactions of small‐molecule inhibitor with the N‐terminal domain of soluble epoxide hydrolase. Support or Funding Information German Research Foundation (DFG; SFB 1039 Teilprojekt A07; Heisenberg‐Professur PR 1405/4‐1)X‐ray structure of N‐terminal domain of soluble epoxide hydrolase in complex with sEH‐phosphatase inhibitor SWE102.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .