Secondary screening of neuroprotective compounds.

<p>Compounds identified through high-throughput screening were administered at 0.1 (white), 1 (light gray), 10 (dark gray), and 100 µM (black) at reoxygenation after 2 hours OGD. Twenty-four hours later, cells survival was measured by TUNEL assay and compared to DMSO vehicle-treated neurons. Compounds providing at least 2-fold increased neuroprotection over vehicle were further investigated. MIA, mianserine hydrochloride, ISO, isoxsuprine hydrochloride, MER, meropenem, MEC, meclofenamic acid, ETI, etilifrine hydrochloride, HAL, haloperidol, MOX, moxonidine, CHL, chlorphenesin carbamate, PRO, prothionamide, EPI, epitiostanol.</p

Dose optimization and time course of administration of neuroprotective compounds.

<p>A. Dose response of compounds administered at reoxygenation after 2(ISO) and etilifrine hydrochloride (ETI) and 200 µM for chlorphenesin carbamate (CHL). Isoxsuprine was significantly more protective at 1 nM compared to 0.01, 0.1, and 100 nM (*, <i>p</i><0.01). Etilifrine was significantly more protective at 1 nM compared to 0.01 and 0.1 nM (*, <i>p</i><0.05). Chlorphenesin carbamate was significantly more neuroprotective at 200 µM compared to all other doses (*, <i>p</i><0.05). B. Time course of administration of compounds at the optimal dose at 0, 15, 30, and 60 minutes after reoxygenation onset. Isoxsuprine and chlorphenesin carbamate demonstrated no decrease in neuroprotection when administered up to 60 minutes after reoxygenation onset. Neuroprotection by etilifrine significantly decreased when administered at 60 minutes versus time 0 (*, <i>p</i><0.01).</p

Neuroprotection by isoxsuprine hydrochloride in an animal stroke model.

<p>A. Representative TTC-stained brain sections showing differences in infarction between animals receiving vehicle or isoxsuprine hydrochloride. B. Effect of isoxsuprine hydrochloride on infarct volume. Isoxsuprine hydrochloride (1 mg/kg, IV) given at the onset of reperfusion after a 90-minute MCAO significantly reduced infarct volume compared to vehicle (137±18 mm³ versus 279±25 mm³), <i>p</i><0.001. Closed circles, vehicle, closed squares, isoxsuprine hydrochloride, n = 7 animals for each group.</p

Neuroprotective compounds identified through high-throughput screening of the Prestwick Chemical Library.

1<p>Compounds previously investigated in models of cerebral ischemia are italicized.</p