3 research outputs found
Proposed Mode of Binding and Action of Positive Allosteric Modulators at Opioid Receptors
Available
crystal structures of opioid receptors provide a high-resolution
picture of ligand binding at the primary (“orthosteric”)
site, that is, the site targeted by endogenous ligands. Recently,
positive allosteric modulators of opioid receptors have also been
discovered, but their modes of binding and action remain unknown.
Here, we use a metadynamics-based strategy to efficiently sample the
binding process of a recently discovered positive allosteric modulator
of the δ-opioid receptor, BMS-986187, in the presence of the
orthosteric agonist SNC-80, and with the receptor embedded in an explicit
lipid–water environment. The dynamics of BMS-986187 were enhanced
by biasing the potential acting on the ligand–receptor distance
and ligand–receptor interaction contacts. Representative lowest-energy
structures from the reconstructed free-energy landscape revealed two
alternative ligand binding poses at an allosteric site delineated
by transmembrane (TM) helices TM1, TM2, and TM7, with some participation
of TM6. Mutations of amino acid residues at these proposed allosteric
sites were found to either affect the binding of BMS-986187 or its
ability to modulate the affinity and/or efficacy of SNC-80. Taken
together, these combined experimental and computational studies provide
the first atomic-level insight into the modulation of opioid receptor
binding and signaling by allosteric modulators
Ligand-Based Discovery of a New Scaffold for Allosteric Modulation of the μ‑Opioid Receptor
With
the hope of discovering effective analgesics with fewer side
effects, attention has recently shifted to allosteric modulators of
the opioid receptors. In the past two years, the first chemotypes
of positive or silent allosteric modulators (PAMs or SAMs, respectively)
of μ- and δ-opioid receptor types have been reported in
the literature. During a structure-guided lead optimization campaign
with ÎĽ-PAMs BMS-986121 and BMS-986122 as starting compounds,
we discovered a new chemotype that was confirmed to display ÎĽ-PAM
or ÎĽ-SAM activity depending on the specific substitutions as
assessed by endomorphin-1-stimulated β-arrestin2 recruitment
assays in Chinese Hamster Ovary (CHO)-ÎĽ PathHunter cells. The
most active ÎĽ-PAM of this series was analyzed further in competition
binding and G-protein activation assays to understand its effects
on ligand binding and to investigate the nature of its probe dependence
Utilization of an Active Site Mutant Receptor for the Identification of Potent and Selective Atypical 5‑HT<sub>2C</sub> Receptor Agonists
Agonism of the 5-HT<sub>2C</sub> receptor
represents one of the most well-studied and clinically proven mechanisms
for pharmacological weight reduction. Selectivity over the closely
related 5-HT<sub>2A</sub> and 5-HT<sub>2B</sub> receptors is critical
as their activation has been shown to lead to undesirable side effects
and major safety concerns. In this communication, we report the development
of a new screening paradigm that utilizes an active site mutant D134A
(D3.32) 5-HT<sub>2C</sub> receptor to identify atypical agonist structures.
We additionally report the discovery and optimization of a novel class
of nonbasic heterocyclic amide agonists of 5-HT<sub>2C</sub>. SAR
investigations around the screening hits provided a diverse set of
potent agonists at 5-HT<sub>2C</sub> with high selectivity over the
related 5-HT<sub>2A</sub> and 5-HT<sub>2B</sub> receptor subtypes.
Further optimization through replacement of the amide with a variety
of five- and six-membered heterocycles led to the identification of
6-(1-ethyl-3-(quinolin-8-yl)-1<i>H</i>-pyrazol-5-yl)Âpyridazin-3-amine
(<b>69</b>). Oral administration of <b>69</b> to rats
reduced food intake in an ad libitum feeding model, which could be
completely reversed by a selective 5-HT<sub>2C</sub> antagonist