Synthesis and biological activity of spin-labeled analogs of biotin, hexamethonium, decamethonium, dichlorisoproterenol, and propranolol.

Spin-labeled analogs of biotin (vitamin H), hexamethonium, decamethonium, dichlorisoproterenol, propranolol, and primaquine containing the nitroxide free radical have been synthesized and tested for biological activity. The four spin-labeled analogs of biotin, 4-biotinamido-2,2,6,6-tetramethyl-1-piperidinyloxy (IV), 3-biotinamido-2,2,5,5-tetramethyl-1-pyrrolidinyloxy (V), 3-biotinamidomethyl-2,2,5,5-tetramethyl-1-pyrrolidinyloxy (VI), and 4-(biotinylglycyl)amino-2,2,6,6-tetramethyl-1-piperidinyloxy (VII), all interacted with avidin, a specific biotin binding protein found in raw egg white, at the same sites as did biotin itself. An unsymmetrical decamethonium spin label (XVIII) in which one of the quaternary methyl groups had been replaced by the 4-(2,2,6,6-tetramethyl-1-piperidinyloxy) moiety was 13 times more potent as an inhibitor of Torpedo californica acetylcholinesterase than the parent drug. The symmetrical decamethonium (XVI) and hexamethonium (XIV) spin labels were 18 and 1.8 times as active as decamethonium in the same assay system. The substitution of the 4-(2,2,6,6-tetramethyl-1-piperidinyloxy) group for the isopropyl groups of beta-adrenergic blocking drugs dichlorisoproterenol and propranolol, to give spin labels XXI and XXII, caused a 45 and 54% reduction, respectively, in the ability of these compounds to inhibit the isoproterenol-stimulated activity of rat fat cell membranes. Finally, modification of primaquine by the introduction of the 4-(2,2,6,6-tetramethyl-1-piperidinyloxy) substituent into the amino group of the butyl side chain completely abolished the ability of the drug to bind to nuclei acids. These results suggest that the incorporation of the nitroxide group into drug molecules may be a useful approach to the synthesis of more specific spin labels for biological systems, such as egg white avidin, acetylcholinesterase, and the beta-adrenergic receptor.