Next, Asim Mahmood will talk about treatment of traumatic brain injury with Vipaloximer, looking at functional recovery in rats. This study was carried out in collaboration with LifeRef Technologies, California, who provided us with this drug, Vipaloximer. No other financial assistance was provided. Traumatic brain injury is a very common problem, just in one single country, not in the United States. There are over a million hospital visits, out of which there are 50,000 deaths and 80,000 cases of long-term morbidity. And all we can do up till now is to remove mass lesions from the brain and provide an optimal milieu in ICU for the brain to recover. There's little available to repair a damaged neural pair. Vipaloximer is a tri-block polypolymer that seals membrane and restores plasma membrane integrity, and it has been used in experimental models of muscular dystrophy, heart failure, and neurodegenerative disorders. It has been reported to attenuate TBI-induced damage by reducing blood-brain barrier damage, brain edema, apoptosis, and mitochondrial dysfunction. It is presently being used in clinical trials of sickle cell disease and heart failure. The drug is FDA-approved. We used a controlled cortical impact model of traumatic brain injury, where the injury is induced over the left cortex with a pneumatic piston, 6 millimeter in diameter, with a velocity of 4 meter per second and 2.5 millimeter compression. This is a standard model which we have used for over two decades. The animals were divided into three groups, a sham group, which underwent sham surgery without injury, and then received the vehicle PBS, that is, saline, and TBI group, the control group, sorry, which received the vehicle that is PBS after an injury, and the experimental group, which were treated with Vipaloximer, 300 milligram per kg, starting two hours post-injury and given via intravenous infusion over 60 minutes. In addition to this group, another set of rats with five per group were used to test the bleeding time, performed one-hour and 24-hour post-drug treatment using a tail transaction bleeding test. 100 functional tests were performed, including MLSS, which was done pre-injury, then on day 1, 4, 7, 14, 21, 28, and 35. It's a composite of motor, sensory, and reflex tests. Neurological function is graded on a scale of 0 to 18, and the higher the score, the greater the deficit. One score is awarded for each abnormal behavior or for lack of tested reflex. This is a standard test of functional analysis. In addition to that, to test the spatial learning, we used Morris Water Maze test, and the animals were tested for five days, starting days 31 and 35, ranging from days 31 to 35 after injury. And the result parameter was the time spent in the correct quadrant. Foot fault tests were also used to measure the sensory motor function, carried pre-injury, then on days 1, 7, 14, 21, 28, and 35 after the injury. Histological analysis were performed with standard H&E tests, as well as immunohistochemistry with CD68, which is a marker of microglia and macrophages, and for GFAP, which is a marker for astrocytes. The medication significantly decreases the volume of total brain tissue loss. This is the sham group compared to the control group, and then the treated group. There was a significant reduction of the volume loss. The treated animals had a significant improvement in the spatial learning as tested by the Morris Water Maze test. This is again the sham group, which has obviously the best results. These are the control animals, and these are the treated animals. It also, the treatment also promoted the functional recovery, tested with the foot fault tests. Vipaloxamol decreased the number of brain microglia, macrophages, and astrocyte activation after the injury. This is the cortex, corpus callosum, and dentate gyrus in sham control and the treated animals. And you can see that the number of microglia and macrophages were decreased in the treated animals. The GFAP area was also reduced, indicating decrease in the activation of the astrocytes. The treatment also reduced brain microthrombosis and hemorrhage in rats. You can see the control in the treated animals, reduction in the hemorrhage. Microthrombosis was decreased. There was no reduction in the body weight with the treatment, which is important. Body weight more or less remained the same in all groups of animals. The administration of Vipaloxamol, it normalized the bleeding time. Vipaloxamol, it's a membrane stabilizer. It repairs by resealing the damaged cell membranes and protects the neuron by suppressing apoptosis. In a stroke model, Vipaloxamol has been shown to increase blood flow in areas of severe and moderate ischemia. Our data also showed that Vipaloxamol decreased astrocyte activation. Astrocytes release pro- and anti-inflammatory cytokines, which can further damage the neurons. The treatment restored coagulation by reducing brain microthrombosis, which can lead to hemodynamic depression and ischemia. Vipaloxamol is neuroprotective by inhibiting megachannel opening. It's a membrane stabilizer. That is the rationale for using it in sickle cell disease. So in conclusion, our study shows that Vipaloxamol treatment after TBI decreases tissue loss, promotes functional recovery, has an anti-inflammatory effect on microglia, macrophages, and astrocytes, normalizes bleeding time, reduces brain hemorrhage and microthrombosis formation, is well-tolerated as indicated by absence of weight loss. So drug has definitely a therapeutic potential, and further studies are essential to take it from the lab to being clinically relevant. Thank you.

Asim Mahmood

traumatic brain injury

Vipaloximer

LifeRef Technologies

TBI treatment