Comparison of intracellular uptake and target protein binding characteristics of two BTK inhibitors with different selectivities
10.12360/CPB202405007
- VernacularTitle:两种不同选择性的BTK抑制剂胞内摄取和靶蛋白结合特点的比较
- Author:
Teng ZHANG
1
;
Ting-Fei TAN
;
Ying-Li ZHAO
;
Gong-Wei HAN
;
Zi-Tong XIA
;
Han-Bing SHI
;
He-Ying LIU
;
Jun-Ping WANG
;
Quan XIA
Author Information
1. 安徽医科大学药学院,安徽合肥 230032
- Keywords:
ibrutinib;
zanubrutinib;
BTK selectivity;
intracellular uptake;
targeted binding characteristics;
hemorrhage
- From:
Chinese Pharmacological Bulletin
2024;40(10):1899-1905
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the intracellular up-take and target protein binding characteristics of two Bruton's tyrosine kinase inhibitors(BTKi)with differ-ent selectivities to provide further insights into the mechanisms of drug off-target-related bleeding risk.Methods Ibrutinib(non-selective BTKi)and za-nubrutinib(selective BTKi)were used as study drugs.After incubation of MEC-1 cells and human platelets with drugs,the cellular thermal shift assay(CETSA)was combined with Western blot to obtain the melting curve and isothermal curve to analyze the binding char-acteristics of the two drugs with the target protein BTK.After incubation of MEC-1 cells and human platelets with drugs,the concentrations of the two drugs were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS)to analyze the intracellular uptake of the two drugs.Results CETSA analysis confirmed that zanubrutinib was more selective for the target protein BTK compared to ibrutinib.LC-MS/MS analysis showed that both drugs were uptaken intracel-lularly by MEC-1 cells and platelets in a concentration-dependent manner.Conclusions While BTKi targe-ting BTK to B lymphocytes exerts therapeutic effects,off-target effects on platelets due to differences in their intracellular uptake,and target-binding characteristics may be one of the reasons for the differences in bleed-ing risk across selective BTKi.
