Microplasmin: a novel thrombolytic that improves behavioral outcome after embolic strokes in rabbits.
OBJECTIVE
It has been proposed that the novel thrombolytic microplasmin may be useful in the treatment of ischemic stroke. In the present study the effects and safety profile of microplasmin were evaluated in 2 rabbit embolic stroke models that have been used successfully to develop tissue plasminogen activator (tPA) as the only Food and Drug Administration-approved treatment for stroke. The rabbit small clot embolic stroke model (RSCEM) and rabbit large clot embolic stroke model (RLCEM) were used to determine the potential neuroprotective properties and safety profile of microplasmin, respectively, after an embolic stroke.


METHODS
Rabbits were embolized by injecting small blood clots (RSCEM) or large blood clots (RLCEM) into the cerebral circulation. For the RSCEM, 126 rabbits were included, with behavioral analysis conducted 24 hours later, allowing for determination of the effective stroke dose (ES50) or clot amount (milligrams) that produces severe neurological deficits in 50% of rabbits. For RLCEM safety study analysis, 47 rabbits were included, with postmortem analyses consisting of assessment of hemorrhage and infarct rate and size. In test animals microplasmin was infused intravenously 60 minutes after embolization, whereas control rabbits were given infusions of the saline/Plasma-Lyte vehicle with all assessments performed in a blinded fashion.


RESULTS
In the RSCEM, a drug is considered neuroprotective if it significantly increases the ES50 compared with the vehicle-treated control group. The ES50 of the vehicle-treated control group 24 hours after embolization was 1.36+/-0.42 mg (n=38). Microplasmin, infused starting 60 minutes after embolization, increased the ES50 to 2.32+/-0.57 (n=21), 1.89+/-0.48 (n=21), 2.81+/-0.55 (n=22), and 1.89+/-0.28 mg (n=24) for the 1-, 2-, 4-, and 8-mg/kg doses, respectively. There was a statistically significant behavioral improvement in the 4-mg/kg dose arm (P=0.040). The microplasmin dose of microplasmin that was statistically significant (4 mg/kg) was subsequently determined to be safe in the RLCEM because it did not increase the incidence of hemorrhages (56%) compared with vehicle-treated rabbits (63%), nor did it significantly alter hemorrhage volume, infarct rate, or infarct volume.


CONCLUSIONS
The present study shows that microplasmin improves behavioral rating scores in the RSCEM when administered 60 minutes after embolization, at a dose that does not increase hemorrhages in the RLCEM. This is in contrast to tPA, which significantly enhances the hemorrhage rate in the RLCEM.
Lapchak
Paul A
PA
Department of Neuroscience, University of California at San Diego, La Jolla, CA 92093-0624, USA. plapchak@ucsd.edu
Araujo
Dalia M
DM
Pakola
Steve
S
Song
Donghuan
D
Wei
Jiandong
J
Zivin
Justin A
JA
D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D002543:Cerebral Hemorrhage / Q000209:etiology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005341:Fibrinolysin / Q000009:adverse effects / Q000627:therapeutic use*; D005343:Fibrinolytic Agents / Q000009:adverse effects / Q000627:therapeutic use*; D007262:Infusions, Intravenous; D020766:Intracranial Embolism / Q000150:complications / Q000188:drug therapy*; D008297:Male; D018696:Neuroprotective Agents / Q000009:adverse effects / Q000627:therapeutic use*; D010446:Peptide Fragments / Q000009:adverse effects / Q000627:therapeutic use*; D011817:Rabbits; D020521:Stroke / Q000150:complications / Q000188:drug therapy*; D016896:Treatment Outcome
D005343:Fibrinolytic Agents; D018696:Neuroprotective Agents; D010446:Peptide Fragments; C054561:microplasmin; D005341:Fibrinolysin