ABSTRACT
Observations that N-Methyl-D-Aspartate (NMDA) antagonists produce symptoms in humans that are similar to those seen in schizophrenia have led to the current hypothesis that schizophrenia might result from NMDA receptor hypofunction. Inhibition of D-amino acid oxidase (DAAO), the enzyme responsible for degradation of D-serine, should lead to increased levels of this co-agonist at the NMDA receptor, and thereby provide a therapeutic approach to schizophrenia. We have profiled some of the preclinical biochemical, electrophysiological, and behavioral consequences of administering potent and selective inhibitors of DAAO to rodents to begin to test this hypothesis. Inhibition of DAAO activity resulted in a significant dose and time dependent increase in D-serine only in the cerebellum, although a time delay was observed between peak plasma or brain drug concentration and cerebellum D-serine response. Pharmacokinetic/pharmacodynamic (PK/PD) modeling employing a mechanism-based indirect response model was used to characterize the correlation between free brain drug concentration and D-serine accumulation. DAAO inhibitors had little or no activity in rodent models considered predictive for antipsychotic activity. The inhibitors did, however, affect cortical activity in the Mescaline-Induced Scratching model, produced a modest but significant increase in NMDA receptor-mediated synaptic currents in primary neuronal cultures from rat hippocampus, and resulted in a significant increase in evoked hippocampal theta rhythm, an in vivo electrophysiological model of hippocampal activity. These findings demonstrate that although DAAO inhibition did not cause a measurable increase in D-serine in forebrain, it did affect hippocampal and cortical activity, possibly through augmentation of NMDA receptor-mediated currents.
Subject(s)
Brain/metabolism , D-Amino-Acid Oxidase/antagonists & inhibitors , Memory, Short-Term/physiology , Psychomotor Agitation/drug therapy , Receptors, N-Methyl-D-Aspartate/metabolism , Animals , Brain/drug effects , Central Nervous System Stimulants/metabolism , Central Nervous System Stimulants/pharmacology , Cyclic GMP/analysis , Cyclic GMP/biosynthesis , D-Amino-Acid Oxidase/metabolism , D-Amino-Acid Oxidase/physiology , Drug Evaluation, Preclinical , Electroencephalography , Habituation, Psychophysiologic/drug effects , Habituation, Psychophysiologic/physiology , Harmaline/metabolism , Hippocampus/drug effects , Hippocampus/metabolism , Male , Maze Learning/drug effects , Maze Learning/physiology , Memory, Short-Term/drug effects , Mescaline/pharmacology , Mice , Miniature Postsynaptic Potentials/drug effects , Miniature Postsynaptic Potentials/physiology , Models, Biological , Models, Chemical , Molecular Targeted Therapy , Motor Activity/drug effects , Motor Activity/physiology , Pruritus/chemically induced , Pruritus/prevention & control , Rats , Rats, Long-Evans , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/drug effects , Sensory Gating/drug effects , Sensory Gating/physiology , Serine/blood , Serotonin Receptor Agonists/pharmacologyABSTRACT
3-Hydroxyquinolin-2(1H)-one (2) was discovered by high throughput screening in a functional assay to be a potent inhibitor of human DAAO, and its binding affinity was confirmed in a Biacore assay. Cocrystallization of 2 with the human DAAO enzyme defined the binding site and guided the design of new analogues. The SAR, pharmacokinetics, brain exposure, and effects on cerebellum D-serine are described. Subsequent evaluation against the rat DAAO enzyme revealed a divergent SAR versus the human enzyme and may explain the high exposures of drug necessary to achieve significant changes in rat or mouse cerebellum D-serine.
Subject(s)
D-Amino-Acid Oxidase/antagonists & inhibitors , Hydroxyquinolines/pharmacology , Hydroxyquinolines/pharmacokinetics , Animals , Cerebellum/metabolism , Crystallography, X-Ray , Drug Discovery , Drug Evaluation, Preclinical , Humans , Hydroxyquinolines/chemical synthesis , Male , Mice , Rats , Rats, Sprague-Dawley , Serine/metabolism , Structure-Activity RelationshipABSTRACT
Explorations in the pyrimidinetrione series of MMP-13 inhibitors led to the discovery of a series of spiro-fused compounds that are potent and selective inhibitors of MMP-13. While other spiro-fused motifs are hydrolytically unstable, presumably due to electronic destabilization of the pyrimidinetrione ring, the spiropyrrolidine series does not share this liability. Greater than 100-fold selectivity versus other MMP family members was achieved by incorporation of an extended aryl-heteroaryl P1'group. When dosed as the sodium salt, these compounds displayed excellent oral absorption and pharmacokinetic properties. Despite the selectivity, a representative of this series produced fibroplasia in a 14 day rat study.
Subject(s)
Matrix Metalloproteinase Inhibitors , Protease Inhibitors/chemistry , Pyrimidines/chemistry , Pyrrolidines/chemistry , Spiro Compounds/chemistry , Animals , Enzyme Stability/drug effects , Matrix Metalloproteinase 13/metabolism , Protease Inhibitors/pharmacology , Pyrimidines/pharmacology , Pyrrolidines/pharmacology , Rats , Spiro Compounds/pharmacology , Structure-Activity RelationshipABSTRACT
Using SAR from two related series of pyrimidinetrione-based inhibitors, compounds with potent MMP-13 inhibition and >100-fold selectivity against other MMPs have been identified. Despite high molecular weights, clogPs, and polar surface areas, the compounds are generally well absorbed and have excellent pharmacokinetic (PK) properties when dosed as sodium salts. In a rat fibrosis model, a compound from the series displayed no fibrosis at exposures many fold greater than its MMP-13 IC50.
Subject(s)
Matrix Metalloproteinase Inhibitors , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Pyrimidinones/chemistry , Animals , Fibrosis/drug therapy , Fibrosis/pathology , Hydroxamic Acids/chemistry , Inhibitory Concentration 50 , Molecular Weight , Rats , Salts/chemistry , Sodium/chemistry , Structure-Activity RelationshipABSTRACT
Through the use of computational modeling, a series of pyrimidinetrione-based inhibitors of MMP-13 was designed based on a lead inhibitor identified through file screening. Incorporation of a biaryl ether moiety at the C-5 position of the pyrimidinetrione ring resulted in a dramatic enhancement of MMP-13 potency. Protein crystallography revealed that this moiety binds in the S(1)(') pocket of the enzyme. Optimization of the C-4 substituent of the terminal aromatic ring led to incorporation of selectivity versus MMP-14 (MT-1 MMP). Structure activity relationships of the biaryl ether substituent are presented as is pharmacokinetic data for a compound that meets our in vitro potency and selectivity goals.
