The Stereoselective Sulfate Conjugation of 4′-Methoxyfenoterol Stereoisomers by Sulfotransferase Enzymes

The presystemic sulfate conjugation of the stereoisomers of 4′-methoxyfenoterol, (R,R′)-MF, (S,S′)-MF, (R,S′)-MF, and (S,R′)-MF, was investigated using commercially available human intestinal S9 fractions, a mixture of sulfotransferase (SULT) enzymes. The results indicate that the sulfation was stereospecific and that an S-configuration at the β-OH carbon of the MF molecule enhanced the maximal formation rates with (S,R′)-MF (S,S′)-MF (R,S′)-MF ≈ (R,R′)-MF, and competition studies demonstrated that (S,R′)-MF is an effective inhibitor of (R,R′)-MF sulfation (IC50 = 60 μM). In addition, the results from a cDNA-expressed human SULT isoform screen indicated that SULT1A1, SULT1A3, and SULT1E1 can mediate the sulfation of all four MF stereoisomers. Previously published molecular models of SULT1A3 and SULT1A1 were used in docking simulations of the MF stereoisomers using Molegro Virtual Docker. The models of the MF-SULT1A3 and MF-SULT1A1 complexes indicate that each of the two chiral centers of MF molecule plays a role in the observed relative stabilities. The observed stereoselectivity is the result of multiple hydrogen bonding interactions and induced conformational changes within the substrate–enzyme complex. In conclusion, the results suggest that a formulation developed from a mixture of (R,R′)-MF and (S,R′)-MF may increase the oral bioavailability of (R,R′)-MF. Chirality 24:796–803, 2012. © 2012 Wiley Periodicals, Inc.1