S(+)-4-(1-Phenylethylamino)quinazolines as inhibitors of human immunoglobulin E synthesis: potency is dictated by stereochemistry and atomic point charges at N-1

• Published in: Journal of medicinal chemistry 2001-08, Vol.44 (18), p.3031-3038
• Main Authors: Berger, M, Albrecht, B, Berces, A, Ettmayer, P, Neruda, W, Woisetschläger, M
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - pharmacology; B-Lymphocytes - metabolism; Blotting, Northern; CD40 Antigens - immunology; Cell Line; Depression, Chemical; Enzyme-Linked Immunosorbent Assay; ErbB Receptors - metabolism; Female; Genes, Reporter; Germ Cells; Humans; Immunoglobulin E - biosynthesis; Immunoglobulin E - genetics; Interleukin-4 - pharmacology; Luciferases - genetics; Luciferases - metabolism; Mice; Mice, Inbred BALB C; Phosphorylation; Promoter Regions, Genetic; Quinazolines - chemical synthesis; Quinazolines - chemistry; Quinazolines - pharmacology; Reproducibility of Results; Spleen - cytology; Stereoisomerism; Structure-Activity Relationship; Tyrosine - metabolism
• ISSN: 0022-2623

Since the pathogenesis of allergic diseases is associated with elevated levels of immunoglobulin E (IgE), we developed a high throughput reporter gene assay in a human B-cell line to screen for low molecular weight IgE inhibitory compounds. Monitoring the IL-4 driven IgE-germline promoter activity (IgE-GLP), we discovered 4-(1-phenylethylamino)qinazolines as potent inhibitors of IgE-germline gene expression. Testing of the individual enantiomers (1, 2) revealed that only the S(+) enantiomer 1 was active. A cell viability assay done in the same cell line in parallel discriminated the dose-dependent inhibition from a general antiproliferative effect. The observed correlation of the inhibitory potencies found in the reporter gene assay with those measured by IgE-ELISA in primary human splenocytes provided evidence that the blockade of IgE synthesis is the direct consequence of IgE-germline gene inhibition, thereby validating the reporter gene assay. Parallel synthesis in solution rapidly provided a series of analogues of compound 1 with modifications in the phenethylamine side chain and the quinazoline core for SAR studies. Increasing the lipophilicity of the arylalkylamine moiety yielded S(+)-4-(1-(2-naphthyl)ethylamino)quinazoline (6) as the most potent inhibitor (IC(50) of 14 nM) while the R(-) enantiomer was again found to be inactive. Within the set of S enantiomers, quantum mechanical calculations revealed that the IgE inhibitory activity can be quantitatively described by the charge at N-1 of the heterocyclic core and to a lesser extent by the molar refractivity. These results demonstrate the importance of electron-deficient fused 4-aminopyrimidines and lipophilic side chains for biological activity. The strong preference for the S configuration of the phenethylamine side chain is remarkable insofar as biological activity for fused 4-(1-phenylethylamino)pyrimidines has been published for the R enantiomers only (EGFR tyrosine kinase inhibition).