The Investigation Of New Anticoagulant ANV-6L15 In The Treatment Of Traumatic Brain Injury Associated Coagulopathy

Objective: At present,there is no effective treatment for coagulopathy caused by traumatic brain injury(TBI).Common anticoagulants are subject to many restrictions due to their poor anticoagulant effect,cumbersome treatment procedures,and many adverse drug reactions.The new anticoagulant ANV-6L15 has a strong anticoagulant effect because they can target the site of injury and specifically inhibit tissue factor(TF)/ ?.However,its anticoagulant effect in TBI is unknown.This experiment attempts to clarify the anticoagulant mechanism of ANV-6L15 in TBI by verifying the anticoagulant effect of ANV-6L15 in vitro and in TBI mice,and exploring the interaction of ANV-6L15 with microparticles(MP)and TF / ?.This study will provide new ideas for the treatment of coagulopathy after TBI.Methods: 1)The anticoagulant and antithrombin production effects of ANV-6L15 in vitro were tested by the clotting time experiment and thrombin generation experiment;2)The anticoagulant effect of ANV-6L15 and its effect on the blood vessels of TBI mice were verified by tail bleeding experiment,clotting time experiment,enzyme linked immunosorbent assay and Evans blue dye extravasation experiment;3)Explore the mechanism of ANV-6L15 inhibiting MP procoagulant effect by flow cytometry and Western Blot technology;4)To study the effect of ANV-6L15 on the coagulation cascade and activated platelets by thrombin generation experiment and flow cytometry technology;5)The effect of ANV-6L15 on the prognosis and neurological dysfunction of TBI mice was studied by survival analysis and neurological dysfunction score.Results: 1)The ANV-6L15 not only inhibits the coagulation of MP/brain-derived microparticles(BDMP)in vitro,but also inhibits thrombin generation induced by them;2)Compared with the TBI group,the TBI + ANV-6L15 group had less bleeding from the tail vein,longer coagulation time,more fibrinogen and less D-dimer production;3)Compared with the TBI group,the Evans blue dye extravasation decreased in the TBI + ANV-6L15 group;4)Flow cytometry and Western Blot results show that ANV-6L15 can tightly bind to MP,and compared with TBI group,TBI + ANV-6L15 group has less thrombin generation,and less factor ? / Xa-positive and fibrinogen-positive MP and activated platelets;5)Compared with the TBI group,the TBI + ANV-6L15 group had smaller modified neurological severity score and higher survival rate.Conclusion: 1)The ANV-6L15 has a significant anticoagulant effect in vitro.2)The ANV-6L15 can inhibit the hypercoagulable state in TBI mice and has a significant anticoagulant effect;3)The ANV-6L15 can improve the permeability of vessels and reduce tissue damage.The results of this study provide new clues for the treatment of coagulopathy after TBI.4)The ANV-6L15 can bind MP and prevent MP-induced coagulation cascade and platelet activation;5)The ANV-6L15 has a significant inhibitory effect on BDMP,platelet microparticles(PMP)and EMP;6)The ANV-6L15 can also inhibit factor ? /Xa.The results of this experiment lay solid theoretical foundation for the treatment of TBI coagulopathy.7)The ANV-6L15 can treat neurological dysfunction and improve the prognosis of TBI mice.The effectiveness and safety of ANV-6L15 in TBI mice indicates that this new anticoagulant is expected to be an ideal candidate for the treatment of TBI associated coagulopathy.