Abstract
Background
Brigatinib (BRG) have been reported to cause hepatic dysfunction-related events, some of which are serious. Idiosyncratic drug-induced liver injury is often immune-mediated and triggered by drugs or their reactive metabolites. BRG contains a piperazine ring and produces iminium derivatives, which are reactive metabolites, in the process of metabolism.However, it is unknown whether BRG and its reactive metabolites activate immune system.
In this study, we examined that BRG and its reactive metabolites can activate inflammasomes of THP-1 cells for the purpose of assessing immune system activation.
Methods
Differentiated THP-1 cells were incubated with BRG (0.3-3 μM) for 24 hours. FLC-4 cells cultured in 3D incubated with BRG (0.3-3 μM) for 7 days. The supernatant of culture medium of FLC-4 cells was added to differentiated THP-1 cells and cultured for 24 h. 1-aminobenzotriazole (ABT, 1 mM) was used to inhibit cytochromes P450 and YVAD (1 μM) was used to inhibit caspase-1 activity. IL-1β in differentiated THP-1 cells was measured using an ELISA kit. Caspase-1 activity in differentiated THP-1 cells was measured using the Caspase-Glo 1 Inflammasome Assay.
Results and Discussion
Incubation of differentiated THP-1 cells with BRG increased the production of IL-1β and caspase-1 activity. Furthermore, incubation of differentiated THP-1 cells with the supernatant from an incubation of FLC-4 cells with BRG increased the production of IL-1β and caspase-1 activity.On the other hand, the increase of the production of IL-1β by BRG was inhibited by adding YVAD to the vulture medium of the THP-1 cells.
These results revealed that BRG within the therapeutic range directly activates the inflammasome of THP-1 cells, whitch suggested that immune activation was involved as a mechanism for the development of BRG induced adverse reactions.
