iii) BTK inhibitors in 16 h ZIP model
Male C57BL/6 mice (10-12 weeks old) were pre-treated with ONO-4059, CNX-774, Olmutinib, LFM-A13, and Acalabrutinib (1 mg·kg-1; p.o.) or vehicle (5% DMSO, 30% cyclodextrin) one hour prior to zymosan challenge (100 ug; i.p.). Compounds where all purchased from SellectChem, as selected because they had varying properties compared to ibrutinib i.e. more selective acalabrutinib, not marketed as a BTK inhibitor but target BTL Olmutinib, more potent ONO-4059 than ibrutinib and structurally different to ibrutinib CNX-774. All compounds also have publication history of being bioactive following oral gavage. Blood was collected by direct cardiac puncture and peritoneal exudate cells were harvested in ice-cold cell PEC harvest buffer (PBS, 5 mM EDTA) 16 h after zymosan challenge.