Study of interaction of carprofen and its enantiomers with human serum albumin--II. Stereoselective site-to-site displacement of carprofen by ibuprofen.

The site-to-site displacement of carprofen, a site II-specific drug, bound to human serum albumin (HSA) by ibuprofen, another site II-specific drug, was qualitatively and quantitatively studied by circular dichroism (CD) and equilibrium dialysis (ED). Carprofen gives rise to different CD spectra at lower (1:1) and higher (3:1) molar ratios to HSA, indicating different mechanisms for the binding of this drug to its high and low affinity sites on HSA. Ibuprofen at a 5:1 molar ratio to HSA displaces carprofen at a molar ratio of 1:1 to HSA from its high affinity binding site (site II) to its low affinity site (site I), as shown by production of the CD spectrum similar to that obtained in the case of the carprofen-HSA complex at a molar ratio 3:1. As revealed by the ED experiments, the free fraction of carprofen at a molar ratio of 1:2 to HSA (2 x 10(-5) M) was not initially increased by the addition of ibuprofen at a lower concentration, but at a higher concentration (6 x 10(-5) M), the free fraction was increased by only 90%. When site I was sufficiently blocked by a site I-specific drug like warfarin or phenylbutazone (6 x 10(-5) M), there was about a 4-fold increase in the free fraction of carprofen caused by ibuprofen. This site-to-site displacement demonstrated by carprofen was found to be stereospecific as indicated by the highest interaction between the S(+)-enantiomers of carprofen and ibuprofen. Moreover, the displacement of carprofen occurred at the azapropazone region rather than the warfarin region of site I on HSA.