The selenium analog of 6-propylthiouracil. Measurement of its inhibitory effect on type I iodothyronine deiodinase and of its antithyroid activity.

6-Propylthiouracil (PTU), a widely used antithyroid drug for the treatment of Graves' disease, is also a potent inhibitor of Type I iodothyronine deiodinase (ID-1). Inhibition of ID-1 was attributed initially to the formation of a mixed disulfide between PTU and a putative cysteine residue at the active site. It has been demonstrated recently that ID-1 is a selenium-containing enzyme, with selenocysteine, rather than cysteine, at the active site. It seemed possible, therefore, that the selenium analog of PTU (PSeU) might be a more potent inhibitor of ID-1 than PTU. To test this possibility, we developed a procedure for the synthesis of PSeU, and we compared PSeU and PTU as inhibitors of ID-1 in a test system containing 125I-rT3, rat liver microsomes, and dithiothreitol. Deiodinase activity was measured by the increase in 125I-iodide. PTU and PSeU were tested at 0.1, 0.3, 1 and 3 microM. Based on results of four separate experiments, the drugs were essentially equipotent as inhibitors of ID-1, although statistical analysis suggested that PSeU may be slightly more potent than PTU. PTU and PSeU were also compared for antithyroid activity in vivo and in vitro. As inhibitors of the catalytic activity of thyroid peroxidase (TPO), the two drugs were essentially equipotent in iodination and guaiacol assays involving measurements made shortly after the addition of H2O2. However, in in vivo experiments with rats, PSeU showed no appreciable inhibition of organic iodine formation in the thyroid, whereas PTU, as expected, was a potent inhibitor. The lack of inhibition of organic iodine formation in vivo by PSeU suggests that, unlike PTU, it is not concentrated by the thyroid gland. In an iodination system in which H2O2 was generated by glucose-glucose oxidase, both PTU and PSeU, when present at 10 microM, acted as reversible inhibitors of iodination. However, when the drug concentration was raised to 50 microM, TPO was inactivated and iodination was irreversibly inhibited. These results suggest that PTU and PSeU inhibit TPO-catalyzed iodination by similar mechanisms. Under the same conditions, the selenium analog of methimazole (another widely used antithyroid drug) does not inactivate TPO. It acts primarily as a reversible inhibitor of TPO-catalyzed iodination.

The selenium analog of methimazole. Measurement of its inhibitory effect on type I 5'-deiodinase and of its antithyroid activity.

Methimazole (MMI), unlike propylthiouracil (PTU) is a poor inhibitor of type I iodothyronine deiodinase (ID-1). Inhibition of the enzyme by PTU was attributed initially to formation of a mixed disulfide between PTU and a cysteine residue at the active site. Presumably, MMI was unable to form a stable mixed disulfide and thus did not inhibit the enzyme. However, it has been demonstrated recently that ID-1 is a selenium-containing enzyme, with selenocysteine, rather than cysteine, at the active site. This observation raised the possibility that the selenium analog of MMI, methyl selenoimidazole (MSeI), might be a better inhibitor of ID-1 than MMI itself, as formation of the Se-Se bond with the enzyme would be expected to occur more readily than formation of the S-SE bond. To test this possibility, we developed a procedure for the synthesis of MSeI and compared MSeI with MMI and PTU for inhibition of ID-1 and for antithyroid activity. For inhibition of ID-1, MMI and MSeI were tested at concentrations of 10-300 microM. No significant inhibition was observed with MMI. MSeI showed slight but significant inhibition only in the 100-300 microM range. PTU, on the other hand, showed marked inhibition at 1 microM. Thus, replacement of the sulfur in MMI with selenium only marginally increases its inhibitory effect on ID-1. As an inhibitor of ID-1, MSeI is much less than 1% as potent as PTU. MMI and MSeI were also compared for antithyroid activity, both in vivo and in vitro. As an inhibitor of the catalytic activity of thyroid peroxidase, MMI was 4-5 times more potent than MSeI in a guaiacol assay, but only twice as potent in an iodination assay. In in vivo experiments with rats, MMI was at least 50 times more potent than MSeI in inhibiting thyroidal organic iodine formation. The relatively low potency of MSeI in vivo suggests that it is much less well concentrated by the thyroid than in MMI.