Discovery and biological evaluation of N-(3-(7-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-4-methyl-2-oxo-2H-pyrimido[4,5-d][1,3]oxazin-1(4H)-yl)phenyl)acrylamide as potent Bruton’s tyrosine kinase inhibitors

• Published in: Acta pharmacologica Sinica 2020-03, Vol.41 (3), p.415-422
• Main Authors: Lai, Meng-zhen, Song, Pei-ran, Dou, Dou, Diao, Yan-yan, Tong, Lin-jiang, Zhang, Tao, Xie, Hua, Li, Hong-lin, Ding, Jian
• Format: Article
• Language: English
• Subjects: Acrylamide; Agammaglobulinaemia Tyrosine Kinase - antagonists & inhibitors; Agammaglobulinaemia Tyrosine Kinase - metabolism; Antineoplastic Agents - chemical synthesis; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; Autoimmune diseases; B-cell lymphoma; B-cell receptor; Biomedical and Life Sciences; Biomedicine; Bruton's tyrosine kinase; Caspase-3; Cell activation; Cell Cycle - drug effects; Cell Proliferation - drug effects; Cyclin D1; Dose-Response Relationship, Drug; Drug Discovery; Drug Screening Assays, Antitumor; G1 phase; Humans; Immunology; Internal Medicine; Kinases; Medical Microbiology; Molecular Docking Simulation; Molecular Structure; Pharmacology/Toxicology; Poly(ADP-ribose) polymerase; Protein Kinase Inhibitors - chemical synthesis; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - pharmacology; Pyrimidines - chemical synthesis; Pyrimidines - chemistry; Pyrimidines - pharmacology; Signal transduction; Structure-Activity Relationship; Targeted cancer therapy; Tumor Cells, Cultured; Tyrosine kinase inhibitors; Vaccine
• ISSN: 1671-4083; 1745-7254
• DOI: 10.1038/s41401-019-0250-8

Bruton’s tyrosine kinase (BTK) is a key component of the B cell receptor (BCR) signaling pathway and plays a crucial role in B cell malignancies and autoimmune disorders; thus, it is an attractive target for the treatment of B cell related diseases. Here, we evaluated the BTK inhibitory activity of a series of pyrimido[4,5-d][1,3]oxazin-2-one derivatives. Combining this evaluation with structure-activity relationship (SAR) analysis, we found that compound 2 exhibited potent BTK kinase inhibitory activity, with an IC 50 of 7 nM. This derivative markedly inhibited BTK activation in TMD8 B cell lymphoma cells and thus inhibited the in vitro growth of the cells. Further studies revealed that compound 2 dose dependently arrested TMD8 cells at G 1 phase, accompanied by decreased levels of Rb, phosphorylated Rb, and cyclin D1. Moreover, following treatment with compound 2 , TMD8 cells underwent apoptosis associated with PARP and caspase 3 cleavage. Interestingly, the results of the kinase activity assay on a small panel of 35 kinases showed that the kinase selectivity of compound 2 was superior to that of the first-generation inhibitor ibrutinib, suggesting that compound 2 could be a second-generation inhibitor of BTK. In conclusion, we identified a potent and highly selective BTK inhibitor worthy of further development.