Irreversible inhibition of BTK kinase by a novel highly selective inhibitor CHMFL-BTK-11 suppresses inflammatory response in rheumatoid arthritis model

Hong Wu,Jin Dong,Jinghua Wang,Wensheng Chen,Qi Zhou ,Kewei Yu ,Yongfei Chen,Nathanael S Gray,Qiong Huang,Beilei Wang,Li Wang,Cheng Chen,Xiaoxiang Li,Qianqian Wang ,Changwei Hu ,Aoli Wang,Yunfei Deng ,Wei Wei,Xiaochuan Liu,Qiaokang Liang ,Jing Liu,Qingsong Liu

doi:10.1038/s41598-017-00482-4

Abstract

BTK plays a critical role in the B cell receptor mediated inflammatory signaling in the rheumatoid arthritis (RA). Through a rational design approach we discovered a highly selective and potent BTK kinase inhibitor (CHMFL-BTK-11) which exerted its inhibitory efficacy through a covalent bond with BTK Cys481. CHMFL-BTK-11 potently blocked the anti-IgM stimulated BCR signaling in the Ramos cell lines and isolated human primary B cells. It significantly inhibited the LPS stimulated TNF-α production in the human PBMC cells but only weakly affecting the normal PBMC cell proliferation. In the adjuvant-induced arthritis rat model, CHMFL-BTK-11 ameliorated the inflammatory response through blockage of proliferation of activated B cells, inhibition of the secretion of the inflammatory factors such as IgG1, IgG2, IgM, IL-6 and PMΦ phagocytosis, stimulation of secretion of IL-10. The high specificity of CHMFL-BTK-11 makes it a useful pharmacological tool to further detect BTK mediated signaling in the pathology of RA.

Highlights

Rheumatoid arthritis (RA), which is characterized by synovial membrane inflammation and causing joint swelling, cartilage and bone destruction, is an autoimmune inflammation disease that affects about 0.5% of human population[1]
Starting from a quinoline-based scaffold, by employment of the structure based irreversible inhibitor design approach[7], we obtained the compound CHMFL-Bruton’s tyrosine kinase (BTK)-11, which displayed an IC50 of 26.82 nM against purified BTK kinase with the ADP-GloTM biochemical assay (Fig. 1b)
The results demonstrated CHMFL-BTK-11 inhibited BTK wt Y551 phosphorylation with an EC50 of 25 nM, while BTK C481S was remarkably resistant to it (EC50: >3 μM) (Fig. 1c and Supplemental Fig. 1)

Summary

Introduction

Rheumatoid arthritis (RA), which is characterized by synovial membrane inflammation and causing joint swelling, cartilage and bone destruction, is an autoimmune inflammation disease that affects about 0.5% of human population[1]. Upon anti-IgM stimulation, which activates the BTK kinase mediated signaling pathway, CHMFL-BTK-11 potently blocked BTK Y223 phosphorylation (less than 100 nM) (Fig. 2a). In the purified human B cells, upon anti-IgM stimulation, BTK Y223 and PLCγ2 Y1217 were potently inhibited by CHMFL-BTK-11 but they were not affected by the reversible version of the compound CHMFL-BTK-12 (Fig. 2b).

Results

Conclusion