Strategy and exploration of identifying drug combinations for
antithrombosis from Xijiao Dihuang Tang formulae as p38MAPK inhibitor
Background and Purpose: Combination therapy has been widely recognized
internationally, however, identifying drug combinations with synergism
is very challenging. We first proposed a strategy for identifying drug
combinations from herbs with mutual reinforcement or assistance in
traditional Chinese medicine (TCM) formulae, and verified it using the
example of Xijiao Dihuang Tang (XDT) , a classical TCM formula.
Experimental Approach: In vitro the smallest half maximal inhibitory
concentration (IC50) values of the candidate ingredients in XDT and
synergistic neuroprotective effect of the drug combinationsin were
studied in injured PC12 and BV2 cell by CCK8 and flow cytometry assay.
The p38MAPK pathway affected by the drug combinations were predicted and
validated using molecular docking method, biolayer interferometry,
western blot and immunoprecipitation assessment. Antithrombotic of the
drug combinations were evaluated in zebrafish thrombosis model in vivo.
Results Two drug combinations with a combination index value of 0.1- 0.5
were obtained, indicating strong synergy. Our results suggested that,
compared to the individual ingredients, the two drug combinations can
strengthen the synergistic neuroprotective effects and have a
significant antithrombotic effect, and not only enhanced the interaction
with on MKK3, MKK6 and p38 in the p38MAPK pathway, but also interrupted
the MKK3-p38 axis to inhibit MKK3-induced activation of p38 MAPK.
Conclusions Our study identified two synergistic drug combinations as
p38MAPK pathway inhibitors to develop antithrombotics and suggests that
provide approaches and effective means for identifying drug combinations
with synergism from the combination of herb-herb with mutual
reinforcement or assistance in TCM formulae for drug combination