NGF (nerve growth factor) and BDNF (brain-derived neurotrophic factor) are protein molecules (MW 26 and 13.6 kDa, respectively) that are neuroprotective in the middle cerebral artery occlusion (MCAO) rat stroke model. Their mechanism of action involves the activation of transcription factor AP-1 that turns on neuronal growth genes. In our ongoing studies we are designing short peptides that mimic some of the properties of full-length neurotrophic factors. We have synthesized a neuroprotective 14-amino acid peptide (CMX-9236) with an N-terminal docosahexaenoic acid (DHA). DHA enhances entry through the blood-brain barrier. Using primary rat brain cortical cultures and a fluorescent assay we found that CMX-9236 can counteract the excitotoxic effects of glutamate or kainate, reversing the intracellular accumulation of Ca(2+) to normal levels. Administration (i.v.) of CMX-9236 post initiation of ischemia reduced the lesion volumes from 178+/-50 to 117+/-55 mm(3) in the temporary rat MCAO model (90 min), and from 216+/-58 to 127+/-57 mm(3) in the permanent (24 h) model for stroke, corresponding to 34+/-28% (P=0.01) and 41+/-19% (P=0.038) reductions of the infarct volumes. Neurological behavior scores showed 57 and 47% improvements for treated temporary and permanent models, respectively. Dose-response studies indicated a 60-fold activation of AP-1 transcription factor in cells treated with 100 ng/ml of the peptide. These studies illustrate that a small peptide can function as a neuroprotective agent and an activator of a beneficial signal transduction pathway.
