Abstract
We report the design and target validation of chimeric peptide EP45, a novel 45 amino acid monomeric dual agonist peptide that contains amino acid sequence motifs present within the blood glucose-lowering agent exendin-4 (Ex-4) and the appetite-suppressing agent PYY(3–36). In a new high-throughput FRET assay that provides real-time kinetic information concerning levels of cAMP in living cells, EP45 recapitulates the action of Ex-4 to stimulate cAMP production via the glucagon-like peptide-1 receptor (GLP-1R), while also recapitulating the action of PYY(3–36) to inhibit cAMP production via the neuropeptide Y2 receptor (NPY2R). EP45 fails to activate glucagon or GIP receptors, whereas for cells that co-express NPY2R and adenosine A2B receptors, EP45 acts in an NPY2R-mediated manner to suppress stimulatory effects of adenosine on cAMP production. Collectively, such findings are remarkable in that they suggest a new strategy in which the co-existing metabolic disorders of type 2 diabetes and obesity will be treatable using a single peptide such as EP45 that lowers levels of blood glucose by virtue of its GLP-1R-mediated effect, while simultaneously suppressing appetite by virtue of its NPY2R-mediated effect.
Highlights
We report the design and target validation of chimeric peptide EP45, a novel 45 amino acid monomeric dual agonist peptide that contains amino acid sequence motifs present within the blood glucose-lowering agent exendin-4 (Ex-4) and the appetite-suppressing agent PYY(3–36)
These two G protein-coupled receptors (GPCRs) activated by EP45 are of major importance to metabolic homeostasis in humans[5,6,7,8,9,10], and they are both established targets for drug discovery research that is relevant to the treatment of type 2 diabetes (T2DM) and obesity[5,6,11,12,13]
We established a new high-throughput fluorescence resonance energy transfer (FRET) assay for detection of cAMP using our HEK293-H188-C24 cell line that stably expresses the cAMP biosensor H188 developed by Klarenbeek and co-workers[30]
Summary
We report the design and target validation of chimeric peptide EP45, a novel 45 amino acid monomeric dual agonist peptide that contains amino acid sequence motifs present within the blood glucose-lowering agent exendin-4 (Ex-4) and the appetite-suppressing agent PYY(3–36). We report the design and target validation of EP45, a chimeric dual agonist peptide that has the surprising ability to act as an agonist at both the glucagon-like peptide-1 receptor (GLP-1R) and the neuropeptide Y2 receptor (NPY2R) These two GPCRs activated by EP45 are of major importance to metabolic homeostasis in humans[5,6,7,8,9,10], and they are both established targets for drug discovery research that is relevant to the treatment of type 2 diabetes (T2DM) and obesity[5,6,11,12,13]. PYY(3– 36) is currently under investigation for use in the treatment of obesity, and it suppresses appetite[4,9,13,24] Interestingly, exaggerated secretion of GLP-1 and PYY(3–36) occurs in some gastric bypass (RYGB) surgery patients, and these alterations correlate with long-term appetite suppression and weight-loss that is associated with such surgery[25]
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