Identifying compounds that modulate interaction of neuronal nitric oxide synthase and its inhibitor, useful e.g. for treating prediabetic states

摘要

Method for detecting compounds (A) that modulate the complexation between neuronal nitric oxide synthase (nNOS) and the inhibitor of nNOS (PIN). nNOS comprises a 1429 amino acid protein, fully defined in the specification, or it variants. Method for detecting compounds (A) that modulate the complexation between neuronal nitric oxide synthase and the inhibitor of nNOS (PIN). A mixture of test compound, PIN and nNOS is incubated under conditions that allow binding between (i) PIN and nNOS and (ii) test compound and either of the proteins, and any variation, relative to a control, in formation of the PIN/nNOS complex is measured. The control value is measured in absence of test compound; in absence of nNOS or PIN; or in presence of a reference inhibitor. A significant variation in complex formation indicates that the test compound binds to one of the proteins. Independent claims are also included for the following: (1) Protein (2), or its fragments of at least 100 amino acids (aa) provided they include aa 269; (2) 104 specific peptides (P), detected by the new process; (3) Nucleic acid that encodes (2), its fragments or P, specifically a 4290 bp rat sequence (1), reproduced; (4) Kit for detecting (A); and (5) Pharmaceutical composition containing (2) or its fragments, or the compound 1-(4-ethoxyphenyl)-5-(5-(benzimidazol-2-ylthio)-fur-2-ylmethylidene) -hexahydropyrimidin-2,4,6-trione.