RESUMEN
The identification, synthesis, and evaluation of a series of naphthoquinone derivatives as selective inhibitors of human arylamine N-acetyltransferase 1 and mouse arylamine N-acetyltransferase 2 are described. The compounds undergo a distinctive color change (red --> blue) upon binding to these human and mouse NAT isoenzymes driven by a proton transfer event. No color change is observed in the presence of functionally distinct but highly similar isoenzymes which are >70% identical. These molecules may be used as sensors to detect the presence of human NAT1 in cell lysates.
Asunto(s)
Arilamina N-Acetiltransferasa/análisis , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/enzimología , Colorimetría/métodos , Isoenzimas/análisis , Animales , Arilamina N-Acetiltransferasa/antagonistas & inhibidores , Femenino , Humanos , Isoenzimas/antagonistas & inhibidores , Ratones , Modelos MolecularesRESUMEN
The identification, synthesis and evaluation of a series of rhodanine and thiazolidin-2,4-dione derivatives as selective inhibitors of human arylamine N-acetyltransferase 1 and mouse arylamine N-acetyltransferase 2 is described. The most potent inhibitors identified have submicromolar activity and inhibit both the recombinant proteins and human NAT1 in ZR-75 cell lysates in a competitive manner. (1)H NMR studies on purified mouse Nat2 demonstrate that the inhibitors bind within the putative active site of the enzyme.
Asunto(s)
Arilamina N-Acetiltransferasa/antagonistas & inhibidores , Neoplasias de la Mama/tratamiento farmacológico , Isoenzimas/antagonistas & inhibidores , Rodanina/síntesis química , Tiazolidinedionas/síntesis química , Animales , Sitios de Unión , Biomarcadores de Tumor/antagonistas & inhibidores , Neoplasias de la Mama/patología , Inhibidores Enzimáticos , Femenino , Humanos , Ratones , Rodanina/farmacología , Tiazolidinedionas/farmacologíaRESUMEN
A series of 3-aryl(pyrrolidin-1-yl)butanoic acids were synthesized using a diastereoselective route, via a rhodium catalyzed asymmetric 1,4-addition of arylboronic acids in the presence of ( R)-BINAP to a crotonate ester to provide the ( S) absolute configuration for the major product. A variety of aryl substituents including morpholine, pyrazole, triazole, imidazole, and cyclic ether were screened in cell adhesion assays for affinity against αvß1, αvß3, αvß5, αvß6, and αvß8 integrins. Numerous analogs with high affinity and selectivity for the αvß6 integrin were identified. The analog ( S)-3-(3-(3,5-dimethyl-1 H-pyrazol-1-yl)phenyl)-4-(( R)-3-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)pyrrolidin-1-yl)butanoic acid hydrochloride salt was found to have very high affinity for αvß6 integrin in a radioligand binding assay (p Ki = 11), a long dissociation half-life (7 h), very high solubility in saline at pH 7 (>71 mg/mL), and pharmacokinetic properties commensurate with inhaled dosing by nebulization. It was selected for further clinical investigation as a potential therapeutic agent for the treatment of idiopathic pulmonary fibrosis.