Neuronal nitric oxide synthase (nNOS) is a target for development of antineurodegenerative agents. Most nNOS inhibitors mimic l-arginine and have poor bioavailability. 2-Aminoquinolines showed promise as bioavailable nNOS inhibitors but suffered from low human nNOS inhibition, low selectivity versus human eNOS, and significant binding to other CNS targets. We aimed to improve human nNOS potency and selectivity and reduce off-target binding by (a) truncating the original scaffold or (b) introducing a hydrophilic group to interrupt the lipophilic, promiscuous pharmacophore and promote interaction with human nNOS-specific His342. We synthesized both truncated and polar 2-aminoquinoline derivatives and assayed them against recombinant NOS enzymes. Although aniline and pyridine derivatives interact with His342, benzonitriles conferred the best rat and human nNOS inhibition. Both introduction of a hydrophobic substituent next to the cyano group and aminoquinoline methylation considerably improved isoform selectivity. Most importantly, these modifications preserved Caco-2 permeability and reduced off-target CNS binding.

Department of Biochemistry University of Texas Health Science Center San Antonio Texas 78384 7760 United States

Department of Chemistry Department of Molecular Biosciences Chemistry of Life Processes Institute Center for Molecular Innovation and Drug Discovery Northwestern University Evanston Illinois 60208 3113 United States

Department of Pediatrics and Center for Applied Genomics 1st School of Medicine Charles University and BIOCEV 121 08 Prague Czech Republic

Departments of Molecular Biology and Biochemistry Pharmaceutical Sciences and Chemistry University of California Irvine California 92697 3900 United States

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