BR4044 Reduces Fibrin Content and Neutrophil Extracellular Traps in Arterial Thrombi

Drug Candidate IV BR4044 delivers nano‐emulsified 3,3’‐Diindolylmethane (DIM) which shows pre‐clinical efficacy in TBI (Dallera CA, et al Neurotherapeutics. 2025 Mar;22(2):e00531). and AIS (Ramakrishna K, et al Curr Neurovasc Res. 2022;19(5):462‐475). DIM binds to, activates, and modulates Aryl Hydrocarbon Receptor (AhR) activity, inhibiting Hexokinase 2 in cytoplasm, and HIF1α transcription in nucleus to prevent Neutrophil Extracellular Trap (NET) formation and Extracellular Vesicle (EV) release following AIS. The purpose of this study was to demonstrate DIM's activity to modify neutrophil activation which contributes to thrombus composition relevant for the success of MT. Using an established mouse model of FeCl3 induced carotid arterial thrombosis, the IV BR4044 DIM drug candidate was shown to modify carotid thrombus composition. H&E and Immunofluorescent stained thrombus specimens were analyzed using 40X scanning and Indica Labs HALO AI‐supported automated cell counting. DIM treatment significantly increased RBC cellularity, reduced Fibrin content (p versus Vehicle = 0.004), reduced Neutrophils (p = 0.036 versus IV Vehicle), and reduced NETs (p = 0.008 versus IV Vehicle). The DIM induced thrombus composition following treatment in mice duplicated the thrombus composition clinically associated with higher first‐pass recanalization and successful thrombectomy. Present results provide translational support for use of IV DIM BR4044 before and during post‐AIS MT while simultaneously providing BBB and neuronal protection. Presently demonstrated neutrophil inhibition are relevant to clinical findings of higher thrombus Neutrophils and NETs seen in poor AIS outcome, failed embolectomy, and thrombolytic resistance. Prior translational studies show improved outcome in AIS models with depletion of neutrophils and NETs where neutrophils are implicated in “No‐Reflow” phenomena. In conclusion BR4044 administration showed significant reduction of fibrin acquisition in arterial thrombi which is expected to translate into clinical benefits for MT in AIS.