Vanadate complexes in serum: a speciation modeling study.

The speciations of two drug candidate ligands, 2-hydroxypyridine-N-oxide (Hhpno) and 2-mercaptopyridine-N-oxide (Hmpno), with vanadate (V(V)) were determined at 25.0 degrees C and 0.20 mol dm(-3) KCl by pH-metric and (51)V-NMR methods. At pH 7.4, the two predominant compounds with both ligands are the VO(2)L(2) and VO(2)L(OH). NH(4)[VO(2)(hpno)(2)] x 3 H(2)O was prepared in solid form, and its crystal structure was determined by X-ray diffraction. The stabilities of the complexes VO(2)L(2) of five drug candidate ligands were compared at pH 7.4. In view of the stability sequence hpno > maltol approximately hdp (Hhdp: 3-hydroxy-1,2-dimethyl-4-pyridinone) >> mpno > picolinic acid, the first two of these ligands were chosen for equilibrium studies with apotransferrin (apoTf) competition. The V(V)-apoTf stability constants (log K(1) = 6.03 +/- 0.10; log K(2) = 5.46 +/- 0.18) determined by (51)V-NMR spectroscopy were confirmed by ultrafiltration. Both methods proved that there seems to be no hydrogencarbonate-vanadate competition for the apoTf anion-binding positions. The other potential high molecular mass V(V) binder in the serum is human serum albumin (HSA). As no interaction was detected by (51)V-NMR spectroscopy or fluorimetry, the binding properties of HSA were quantified on the basis of literature data. As a final conclusion, speciation modeling calculations suggest that, under serum conditions, apoTf is probably the primary metal ion binder, even in the presence of the most stable V(V) carrier ligands hpno and maltol and HSA plays a negligible role in V(V) binding.