Conformation of the trypanocidal pharmaceutical suramin in its free and bound forms: transferred nuclear overhauser...

Suramin is a lead compound for treatment of cancer, HIV, and trypanosomiasis. The conformations of suramin in its free form and bound to phosphoglycerate kinases from T. brucei and S. cerevisae, have been studied in aqueous solutions using nuclear Overhauser effect (NOE) and transferred NOE NMR spectroscopies. The NOE data of the free drug can be accommodated by a model in which many of the single bonds of suramin are unrestricted at room temperature, consistent with molecular mechanics calculations. The angle between the naphthalene ring and the adjoining amide is essentially locked by a strong amide-sulfonate hydrogen bond into one preferred conformation. Another degree of freedom near the termini of the molecule has a rather pronounced preference, and a third exhibits a nearly perpendicular arrangement between the amide and adjacent aromatic ring. The other two degrees of freedom have weaker preferences. Molecular mechanics calculations using AMBER force field and charges on amides and sulfonates obtained from semiempirical or ab initio calculations reproduced the extent of nonplanarity but not the detailed preferences. 13C spin-lattice relaxation, proton NOE, and light-scattering measurements for free suramin indicate that the correlation time of the molecule is approximately 3 ns at 5 mM concentration, suggesting that suramin is multimeric. Lowering the temperature to 5 degrees C causes a dramatic broadening of all of the resonances in the NMR spectra of 5 mM suramin. This broadening probably is associated with further aggregation into micelles. Suramin is monomeric at 0.5 mM and room temperature, and the NOE cross-relaxation rate constants are close to the cancellation condition for a 500 MHz proton frequency; this concentration is typical of blood serum concentrations when the drug is utilized in humans. Changes in the conformational preferences for terminal degrees of freedom are observed in the bound states of suramin based upon the transferred NOE data. The data for the bound state cannot be accommodated by a symmetric conformer. Analysis of the transferred NOESY buildup curves indicates complex kinetics of binding, probably involving an electrostatically bound encounter complex. Despite the weak binding constant, the buildup curves cannot be treated as population-weighted averages of the free and bound cross-relaxation rates, and therefore complete relaxation-exchange matrix analysis has been performed to simulate the data sets.