Substituted isoquinolines and quinazolines as potential antiinflammatory agents. Synthesis and biological evaluation of inhibitors of tumor necrosis factor alpha

A series of isoquinolin-1-ones and quinazolin-4-ones and related derivative s were prepared and evaluated for their ability to inhibit tumor necrosis f actor alpha (TNF alpha) production in human peripheral blood monocytes stim ulated with bacterial lipopolysaccharide (LPS). In an effort to optimize th e TNF alpha inhibitory activity, a homologous series of N-alkanoic acid est ers was prepared. Several electrophilic and nucleophilic substitutions were also carried out. Alkanoic acid esters of four carbons were found to be op timum for activity in both the isoquinoline and quinazoline series. Ring su bstituents such as fluoro, bromo, nitro, acetyl, and aminomethyl on the iso quinoline ring resulted in a significant loss of activity. Likewise, simila r groups on the quinazoline ring also reduced inhibitory activity. However, the 6- and 7-aminoquinazoline derivatives, 75 and 76, were potent inhibito rs, with IC50 values in the TNF alpha in vitro assay of approximately 5 mu M for each. An in vivo mouse model of pulmonary inflammation was then used to evaluate promising candidate compounds identified in the primary in vitr o assay. Compound 75 was selected for further study in this inhalation mode l, and was found to reduce the level of TNF alpha in brochoalveolar lavage fluid of LPS-treated mice by about 50% that of control mice. Thus, compound s such as 75, which can effectively inhibit proinflammatory cytokines such as TNF alpha in clinically relevant animal models of inflammation and fibro sis, may have potential as new antiinflammatory agents. Finally, a quinazol ine derivative suitable to serve as a photoaffinity radiolabeled compound w as prepared to help identify the putative cellular target(s) for these TNF alpha inhibitors.