Screening And Anti-cerebral Ischemia-reperfusion Injury Effect Of A-FABP Targeting Small Molecule Inhibitors

Ischemic stroke is one of the diseases which threatened human life and reduce the quality of life of patients.After the occurrence of ischemic stroke,the main feature is the damage of the blood-brain barrier.The damaged integrity of the blood-brain barrier can lead to increasing paracellular permeability so that toxins,various immune cells and inflammatory factors enter the brain.The main treatment method in the acute phase of ischemic stroke（within 3.5 to 8 hours）is to dredge blood vessels,so that ischemic brain tissue can restore blood flow in time.A large number of animal and clinical evidences have shown that the vascular recanalization after stroke can cause the cerebral ischemia-reperfusion injury,resulting in the destruction of blood-brain barrier function and increasing nerve cell death,patient mortality and the increased risk of hemorrhagic transformation.Therefore,the development of drugs which can control or alleviate the above-mentioned side effects of vascular recanalization therapy will increase the benefit of vascular recanalization therapy and improve the prognosis of ischemic stroke patients.Adipocyte-type fatty acid binding protein（A-FABP）is an adipokine which affects various metabolic diseases.Since A-FABP has dual functions of fatty acid transport and promotion of inflammation,it is considered to be a key molecule in regulating metabolic function.Recent studies have shown that A-FABP is highly expressed after the cerebral ischemic injury to exacerbate the cerebral ischemia-reperfusion injury,so it is a potential drug target for ischemic stroke However,no A-FABP inhibitor has yet been approved for clinical trials..From the perspective of saving time and cost in drug development,this study adopts the strategy of " discover a new drug is to start with an old one ",through a combination of machine learning and molecular docking,aiming at screening potential A-FABP inhibitors in the FDA-approved drug database,and combining in vitro and in vivo experiments to verify the inhibitory activity of the compounds.The main research contents and therapeutic effect are divided into the following aspects:（1）A naive Bayesian classification model combined with molecular docking model develop a virtual screening method for A-FABP inhibitors,and from FDAapproved drugs（about 2600 types）.Four compounds were screened out which they were Cobimetinib,Larotrectinib,Pantoprazole,and Vildagliptin.（2）Cobimetinib has a good inhibitory effect on the pro-inflammatory function of AFABP in mouse primary macrophages and mouse macrophage cell line RAW 264.7 cells.（3）We used a mouse cerebral ischemia-reperfusion model to verify the therapeutic effect of Cobimetinib.It was found that MCAO model mice could increase the content of A-FABP in serum,and Cobimetinib had no obvious effect on cerebral ischemia-reperfusion injury.Combined with the specific inhibition of MEK by Cobimetinib and the complexity of the effect on the body,its therapeutic effect on ischemic stroke needs further evaluation.In conclusion,the study presents a new method for virtual screening of potential A-FABP inhibitors,further examining the possibility of candidate compounds as novel A-FABP inhibitors,and it finally confirm that Cobimetinib can act as a potential novel A-FABP inhibitor.