Abstract
The seven-transmembrane receptor Smoothened (Smo) is the major component involved in signal transduction of the Hedgehog (Hh) morphogens. Smo inhibitors represent a promising alternative for the treatment of several types of cancers linked to abnormal Hh signaling. Here, on the basis of experimental data, we generated and validated a pharmacophoric model for Smo inhibitors constituted by three hydrogen bond acceptor groups and three hydrophobic regions. We used this model for the virtual screening of a library of commercially available compounds. Visual and structural criteria allowed the selection of 20 top scoring ligands, and an acylthiourea, N-(3-benzamidophenylcarbamothioyl)-3,4,5-trimethoxybenzamide (MRT-10), was identified and characterized as a Smo antagonist. The corresponding acylurea, N-(3-benzamidophenylcarbamoyl)-3,4,5-trimethoxybenzamide (MRT-14), was synthesized and shown to display, in various Hh assays, an inhibitory potency comparable to or greater than that of reference Smo antagonists cyclopamine and N-((3S,5S)-1-(benzo[d][1,3]dioxol-5-ylmethyl)-5-(piperazine-1-carbonyl)pyrrolidin-3-yl)-N-(3-methoxybenzyl)-3,3-dimethylbutanamide (Cur61414). Focused virtual screening of the same library further identified five additional related antagonists. MRT-10 and MRT-14 constitute the first members of novel families of Smo antagonists. The described virtual screening approach is aimed at identifying novel modulators of Smo and of other G-protein coupled receptors.
Footnotes
↵
The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.This work was supported by La Ligue contre le Cancer (Comité des Yvelines); l'Institut National du Cancer; l'Association pour la Recherche contre le Cancer [Grant 3850], l'Association Française contre les Myopathies [Grant 12181]; the French Agence Nationale de la Recherche [Grant 07-physio-010-04]; and The Neuropole de Recherche Francilien [Doctoral Fellowship 248890].
Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.110.065102.
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ABBREVIATIONS:
- Hh
- Hedgehog
- Ptc
- Patched
- GPCR
- G protein-coupled receptor
- Smo
- Smoothened
- BCC
- basal cell carcinoma
- Cur61414
- N-((3S,5S)-1-(benzo[d][1,3]dioxol-5-ylmethyl)-5-(piperazine-1-carbonyl)pyrrolidin-3-yl)-N-(3-methoxybenzyl)-3,3-dimethylbutanamide
- Z″″
- N-(4-chloro-3-(6-(dimethylamino)-1H-benzo[d]imidazol-2-yl)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide
- SAG
- Smoothened agonist
- DMSO
- dimethyl sulfoxide
- HEK
- human embryonic kidney
- IP
- inositol phosphates
- ShhN
- N-terminal fragment of human Sonic hedgehog
- AP
- alkaline phosphatase
- PBS
- phosphate-buffered saline
- DAPI
- 4,6-diamidino-2-phenylindole
- BC
- Bodipy-cyclopamine
- HBA
- hydrogen-bond acceptor
- HY
- hydrophobic
- MRT-24
- N-(3-benzamidophenylcarbamothioyl)-3,4-dimethoxybenzamide
- MRT-29
- 4-methoxy-N-(3-(2-methylbenzamido)phenylcarbamothioyl)-3-nitrobenzamide
- MRT-31
- N-(3-(3-biphenylcarbonylthioureido)phenyl)furan-2-carboxamide
- MRT-39
- N-(3-(3-(3,4-dimethoxybenzoyl)thioureido)phenyl)furan-2-carboxamide
- MRT-42
- N-(5-(benzo[d]thiazol-2-yl)-2-methylphenylcarbamothioyl)-3,4-dimethoxybenzamide.
- Received March 26, 2010.
- Accepted July 27, 2010.
- Copyright © 2010 The American Society for Pharmacology and Experimental Therapeutics
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