Summary Assay for Selective Agonists of NTR1
Addiction leading to abuse should be treatable by pharmacological approaches, and programs that identify new drugs to treat methamphetamine abuse address an immediate goal of the National Institute on Drug Abuse (NIDA) that new approaches are needed for treating METH addiction. Currently, small molecule drug-like compounds are not available for treating METH abuse. Neurotensin receptor 1 (NTR1) more ..
Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG)
Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA)
Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1 R03 MH089653-01
Assay Provider: Dr. Lawrence Barak, Duke University Medical Center
Addiction leading to abuse should be treatable by pharmacological approaches, and programs that identify new drugs to treat methamphetamine abuse address an immediate goal of the National Institute on Drug Abuse (NIDA) that new approaches are needed for treating METH addiction. Currently, small molecule drug-like compounds are not available for treating METH abuse. Neurotensin receptor 1 (NTR1) peptide agonists produce behaviors that are exactly opposite to the psychostimulant effects observed with methamphetamine abuse, such as hyperactivity, neurotoxicity, psychotic episodes, and cognitive deficits, and repeated administrations of NTR1 agonists do not lead to the development of tolerance . These data form the basis of the idea that neurotensin receptors are valid targets for antagonizing drug seeking behaviors and preventing relapses. We propose to identify novel small molecule neurotensin receptor agonists by screening libraries of compounds using a primary assay based upon the ability of a b-arrestin fluorescent reporter to directly recognize the activated state of the NTR1.
This AID summarizes SBCCG's probe development efforts to identify agonists of the neurotensin receptor 1.
From the primary high-content imaging based HTS (AID 493036) and subsequent hit confirmation (AID 504550) several hit compounds were identified. Compounds that confirmed were further tested in full dose response (AID 504549). Hits were clustered into scaffolds and SAR studies were performed on compounds from several scaffolds reconfirming potency and allowing for structure-activity refinement (AID 602356, 602426). Compounds were further tested for selectivity against the related neurotensin receptor 2 (AID 588495, 602334) and the unrelated GPR35 receptor (AID 588503). Selected compounds were further profiled in a NTR1 calcium mobilization assay (AID 602466, 602459), a NTR1 b-arrestin assay (AID 602427) based on enzyme fragment complementation, and a cytotoxicity screen (AID 602482). Successful prosecution of the project led to the nomination of two low, single-digit micromolar, small molecule, non-peptide chemical agonist probes for the neurotensin-1 receptor, with submission of a probe report for ML301 (MLS004254788) and ML314 (MLS004254796). These probes appear to exhibit divergent GPCR pathway bias with ML301 displaying full agonist behavior in b-arrestin pathway based assays as well as a calcium flux assay, while ML314 displays full agonist behavior in b-arrestin pathway based assays, but no activity in a calcium mobilization assay.
Details of protocols, compound structures, and results from the original assays can be found in PubChem at the respective AIDs listed below.
Biology of neurotensin: revisited study. Katsanos GS, Anogianaki A, Castellani ML, Ciampoli C, De Amicis D, Orso C, Pollice R, Vecchiet J, Tete S, Salini V, Caraffa A, Patruno A, Shaik YB, Kempuraj D, Doyle R, Antinolfi PL, Cerulli G, Conti CM, Fulcheri M, Neri G, Sabatino G. Int J Immunopathol Pharmacol. 2008 Apr-Jun;21(2):255-9
Biochemical and pharmacological profile of a potent and selective nonpeptide antagonist of the neurotensin receptor.
D Gully, M Canton, R Boigegrain, F Jeanjean, J C Molimard, M Poncelet, C Gueudet, M Heaulme, R Leyris, A Brouard, et al.
Proc Natl Acad Sci U S A. 1993 January 1; 90(1): 65-69.
Neurotensin receptor antagonists and therapeutical perspectives.
Gully D, Maffrand JP, Soubrie P, Rostene W, Kitabgi P, Le Fur G.
Therapie. 1995 Jan-Feb;50(1):5-7.
Neuropharmacological profile of non-peptide neurotensin antagonists.
Gully D, Jeanjean F, Poncelet M, Steinberg R, Soubrie P, Le Fur G, Maffrand JP. Fundam Clin Pharmacol. 1995;9(6):513-21. Review.
Use of nonpeptide antagonists to explore the physiological roles of neurotensin. Focus on brain neurotensin/dopamine interactions.
Rostene W, Azzi M, Boudin H, Lepee I, Souaze F, Mendez-Ubach M, Betancur C, Gully D. Ann N Y Acad Sci. 1997 Apr 24;814:125-41. Review.
Please see pertinent AIDs: 493036, 504549, 504550, 588495, 588503, 602334, 602356, 602426, 602427, 602459, 602466, 602482
Probe molecules are defined as the positives of this assay and assigned a score of 100. Two compounds have been identified as probes,
ML314 - SID 134225039
ML301 - SID 126723249 & SID 104164387
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 2
MLP Probe ML# for probe 1: ML301
PubChem Substance ID (SID) for probe 1: 126723249
PubChem Compound ID (CID) for probe 1: 49837912
IC50/EC50 (nM) for probe 1: 1890
Target for probe 1: ntr1 (gi: 110611243)
Anti-target for probe 1: ntr2
Fold selectivity for probe 1: 40
Grant number for probe 1: 1 R03 MH089653-01
MLP Probe ML# for probe 2: ML314
PubChem Substance ID (SID) for probe 2: 134225039
PubChem Compound ID (CID) for probe 2: 53245590
Probe type for probe 2: Agonist
IC50/EC50 (nM) for probe 2: <4140
Target for probe 2: NTR1 (gi: 110611243)
Disease relevance for probe 2: Addiction
Anti-target for probe 2: NTR2
Fold selectivity for probe 2: 20
Grant number for probe 2: MH089653-01
PubMed Publication ID (PMID) for probe 1: 24611085
PubMed Publication ID (PMID) for probe 2: 24611085
Data Table (Concise)