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Volumn 17, Issue 16, 2007, Pages 4464-4470

Identification of a novel series of benzimidazoles as potent and selective antagonists of the human melanocortin-4 receptor

Author keywords

Antagonist; Benzimidazole; Melanocortin 4 receptor

Indexed keywords

BENZAMIDINE; BENZIMIDAZOLE DERIVATIVE; MCL 0129; MELANOCORTIN 4 RECEPTOR; ML 253764; PIPERAZINEBENZYLAMINE; RECEPTOR BLOCKING AGENT; UNCLASSIFIED DRUG;

EID: 34447310285     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2007.06.010     Document Type: Article
Times cited : (26)

References (22)
  • 13
    • 34447313153 scopus 로고    scopus 로고
    • note
    • 125I]NDP-α-MSH was separated by filtration through GF/C glass fibre filters presoaked with 0.1% (w/v) PEI. Filters were washed three times with 50 mM Tris-HCl, pH 7.4, at 0-4 °C and assayed for radioactivity using Perkin-Elmer Topcount counter. Binding data were analysed by computer-assisted non-linear regression analysis (XL fit; IDBS).
  • 17
    • 34447344256 scopus 로고    scopus 로고
    • ®) was used instead of N,N′-dicyclohexylcarbodimide to facilitate by-products removal.
  • 21
    • 34447298118 scopus 로고    scopus 로고
    • note
    • Cyclic AMP assay: Intracellular cyclic AMP (cAMP) levels were determined by an electro-chemiluminescence (ECL) assay (Meso Scale Discovery, MSD). CHO-K1 cells stably expressing the human MC4 receptors were suspended in RMPI 1640 containing 0.5 mM IBMX and 0.2% protein cocktail (MSD). They were dispensed (7000 cells/well) in multi-array plates containing integrated carbon electrodes and coated with anti-cAMP antibody. Concentration-response experiments of NDP-α-MSH were carried out in the presence of increasing concentrations of the tested compound by incubating the cells for 40 min at 37 °C. Then, the cells were lysed and 2.5 nM TAG ruthenium-labelled cAMP was added. After 90 min, cAMP levels were determined by ECL detection using sector imager 6000 reader (MSD). cAMP data were analysed by computer-assisted non-linear regression analysis (XL fit; IDBS). The Kb values were determined by Schild regression analysis.
  • 22
    • 34447334582 scopus 로고    scopus 로고
    • note
    • Fed male Sprague-Dawley rats were dosed in groups of three animals (for each kinetic point). A solution of 4.5 μmol/kg in 1 ml/kg of a 10% DMA/90% propanediol solution was given by intravenous administration via the penile vein. Then, rats were deeply anaesthetised under Isoflurane® and blood was taken from the vena cava (at 1, 15, 30, 60, 90 and 120 min) and centrifuged (2000g, 15 min, 4 °C). The supernatants were mixed with cold acetonitrile to precipitate the proteins. After a further centrifugation (2000g, 15 min, 4 °C), the samples were analysed by HPLC (UV detection) or frozen at - 80° pending analysis. After blood withdrawal, tissues were rapidly washed by intracardiac perfusion with 50 ml of saline. The brains were then removed, frozen in liquid nitrogen and weighed. After mixing the brains with methanol for 40 s with an Ultraturax®, brain homogenates were centrifuged (25,000g, 15 min, 4 °C) and the supernatants were analysed by HPLC.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.