Prostaglandin E2 (PGE2) is a lipid autacoid, which is an oxidative metabolite of arachidonic acid synthesized by cyclooxygenase and prostaglandin E synthases. PGE2 is proposed to mediate in part the pathophysiology of hypertensive and diabetic kidney diseases. To probe the structure and function of a cell surface receptor for PGE2, single amino acid substitutions of the mouse EP3 receptor were generated, each having a single cysteine-to-alanine missense mutation. These studies demonstrated a critical disulfide bond between the first and second extracellular loops of mouse EP3 at position C107 and C184. Receptors substituted at either of these cysteines had attenuated expression and were functionally inactive. To study EP1 and EP3 receptors in mouse models of chronic kidney disease, selective antagonists of EP1 and/or EP3 were synthesized and characterized. A 4-chlorophenylsulfonamide was a functional antagonist of EP1 and EP3, lacked the off-target activity of the lead antagonist, and was efficiently metabolized in vitro to a small subset of metabolites. Subcutaneous injection of this compound prolonged plasma exposure of this antagonist sufficiently to maintain receptor coverage with once daily injection. Subcutaneous injection of this antagonist significantly blunted the vasopressor response of EP1 and EP3 agonists in mice. Thus, this represents a novel, selective EP1 and EP3 antagonist that is bioavailable in mice and suitable for use in studies of chronic kidney disease.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-11092012-132438 |
| Date | 21 November 2012 |
| Creators | Downey, Jason Duane |
| Contributors | Roy Zent, Richard Breyer, Vsevolod Gurevich, Lawrence Marnett, Craig Lindsley |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-11092012-132438/ |
| Rights | restricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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