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CELLULAR AND MOLECULAR

-Aminoazaheterocyclic-Methylglyoxal Adducts Do Not Inhibit Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channel Activity

Departments of Medicine and Physiology, University of California, San Francisco, California (N.D.S., W.N., A.S.V.); and Laboratorio di Genetica Molecolare, Istituto Giannina Gaslini, Genova, Italy (O.Z.-M., L.J.V.G.)

Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential applications in the therapy of secretory diarrheas and polycystic kidney disease. In a recent study, several highly polar -aminoazaheterocyclic-methylglyoxal adducts were reported to reversibly inhibit CFTR chloride channel activity with IC₅₀ values in the low picomolar range (J Pharmacol Exp Ther 322:1023–1035, 2007), more than 10,000-fold better than that of thiazolidinone and glycine hydrazide CFTR inhibitors previously identified by high-throughput screening. In this study, we resynthesized and evaluated the -aminoazaheterocyclic-methylglyoxal adducts reported to have high CFTR inhibition potency (compounds 5, 7, and 8). We verified that the reported synthesis procedures produced the target compounds in high yield. However, we found that these compounds did not inhibit CFTR chloride channel function in multiple cell lines at up to 100 µM concentration, using three independent assays of CFTR function including short-circuit current analysis, whole-cell patch-clamp experiments, and yellow fluorescence protein-fluorescence quenching. As positive controls, approximately 100% of CFTR inhibition was found by thiazolidinone and glycine hydrazide CFTR inhibitors. Our data provide direct evidence against CFTR inhibition by -aminoazaheterocyclic-methylglyoxal adducts.

Address correspondence to: Dr. Alan S. Verkman, Departments of Medicine and Physiology, 1246 Health Sciences East Tower, University of California, San Francisco, CA 94143-0521. E-mail: Alan.Verkman{at}ucsf.edu