IMM-H004, A New Coumarin Derivative, Improved Focal Cerebral Ischemia via Blood-Brain Barrier Protection in Rats

J Stroke Cerebrovasc Dis. 2017 Oct;26(10):2065-2073. doi: 10.1016/j.jstrokecerebrovasdis.2016.11.121. Epub 2017 Jun 29.

Abstract

Objective: IMM-H004 (7-hydroxy-5-methoxy-4-methyl-3-[4-methylpiperazin-1-yl]-2H-chromen-2-one) is a novel coumarin derivative that showed better effect in improving global cerebral ischemia in rats. However, the effects and mechanisms in focal cerebral ischemia were not clear. Blood-brain barrier (BBB) protection is a vital strategy for the treatment of cerebral ischemia. This study is to investigate whether IMM-H004 improves brain ischemia injury via BBB protection.

Methods: Focal brain ischemia model was induced by middle cerebral artery occlusion for 1 hour and reperfusion (MCAO/R) for 24 hours in rats. IMM-H004 (1.5, 3, 6 mg/kg) and edaravone (positive drug, 6 mg/kg) were administered after 5 minutes of occlusion. Neurological score and TTC staining were used to evaluate the effect of IMM-H004. Evans Blue (EB) staining and electron microscopy were used to assess BBB permeability. Western blot, reverse transcription-polymerase chain reaction, and immunohistochemistry were used to detect the expression of BBB structure-related proteins.

Results: Compared with the model group, IMM-H004 in the focal brain ischemia model improved neurological function and reduced cerebral infarction size and edema content. IMM-H004 sharply reduced the EB content and alleviated BBB structure. In addition, IMM-H004 increased the level of zonula occludens (ZO-1) and occluding, decreased the level of aquaporin 4 and matrix metalloproteinase 9, either in cortex or in hippocampus. And all of these changed were related to BBB protection.

Conclusion: IMM-H004 improved cerebral ischemia injury via BBB protection. For a potential therapy drug of cerebral ischemia, IMM-H004 merits further study.

Keywords: Cerebral ischemia; IMM-H004; blood–brain barrier; coumarin derivative; electron microscopy.

MeSH terms

  • Animals
  • Aquaporin 4 / metabolism
  • Blood-Brain Barrier / drug effects*
  • Blood-Brain Barrier / metabolism
  • Blood-Brain Barrier / ultrastructure
  • Brain Edema / metabolism
  • Brain Edema / pathology
  • Brain Edema / prevention & control
  • Capillary Permeability / drug effects*
  • Cerebral Cortex / drug effects
  • Cerebral Cortex / metabolism
  • Cerebral Cortex / pathology
  • Coumarins / pharmacology*
  • Cytoprotection
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Hippocampus / drug effects
  • Hippocampus / metabolism
  • Hippocampus / pathology
  • Infarction, Middle Cerebral Artery / metabolism
  • Infarction, Middle Cerebral Artery / pathology
  • Infarction, Middle Cerebral Artery / prevention & control*
  • Male
  • Matrix Metalloproteinase 9 / metabolism
  • Neuroprotective Agents / pharmacology*
  • Occludin / metabolism
  • Rats, Sprague-Dawley
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control*
  • Time Factors
  • Zonula Occludens-1 Protein / metabolism

Substances

  • Aqp4 protein, rat
  • Aquaporin 4
  • Coumarins
  • IMM-H004
  • Neuroprotective Agents
  • Occludin
  • Ocln protein, rat
  • Tjp1 protein, rat
  • Zonula Occludens-1 Protein
  • Matrix Metalloproteinase 9
  • Mmp9 protein, rat