A novel synthetic chemical entity (UPEI‐800) is neuroprotective in vitro and in an in vivo rat model of oxidative stress

• Published in: Clinical and experimental pharmacology & physiology 2017-10, Vol.44 (10), p.993-1000
• Main Authors: Saleh, Tarek M., Saleh, Monique C., Connell, Barry J., Kucukkaya, Inan, Abd‐El‐Aziz, Alaa S.
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Antioxidants; Antioxidants - metabolism; APPC; Cell culture; Cell Death - drug effects; Cerebral blood flow; Chemistry Techniques, Synthetic; Covalence; Deprivation; Glucose; Glucose - metabolism; In vivo methods and tests; Intracellular Space - drug effects; Intracellular Space - metabolism; ischaemia–reperfusion injury; Ischemia; Lipoic acid; mixed neuronal culture; Neuronal-glial interactions; Neurons - drug effects; Neurons - metabolism; Neurons - pathology; Neuroprotection; Neuroprotective Agents - chemical synthesis; Neuroprotective Agents - chemistry; Neuroprotective Agents - pharmacology; Occlusion; Organic chemistry; Oxidative stress; Oxidative Stress - drug effects; Oxygen; Oxygen - metabolism; Pretreatment; Pyrazole; Pyrazoles - chemical synthesis; Pyrazoles - chemistry; Pyrazoles - pharmacology; Rats; Reperfusion; Reperfusion Injury - pathology; Rodents; stroke
• ISSN: 0305-1870; 1440-1681
• DOI: 10.1111/1440-1681.12785

Summary In this study, we tested a novel synthetic pyrazole‐containing compound, 5‐amino‐1‐phenyl‐1H‐pyrazole‐4‐carbonitrile (APPC), as an antioxidant in both in vitro and in vivo models of oxidative stress. In addition, the utility of covalently combining APPC with another well‐established antioxidant, lipoic acid (LA), was also tested in both models. The in vitro results demonstrated that pretreatment with APPC in a mixed neuronal‐glial culture exposed to oxygen‐glucose deprivation (OGD) followed by reoxygenation‐refeeding, resulted in significant neuroprotection at concentrations between 2.5 to 25 μmol/L. In contrast, LA was not neuroprotective following OGD alone or following reoxygenation‐refeeding. However, the synthetic covalent combination of APPC with LA, named “UPEI‐800”, resulted in significant neuroprotection at concentrations between 0.027 and 2.7 μmol/L (100‐fold more potent than APPC alone), an effect shown to be correlated with increased cellular antioxidant capacity. Further, in an in vivo model of ischaemia‐reperfusion injury following transient occlusion of the middle cerebral artery (tMCAO), both APPC (0.1 and 1.0 mg/kg) and UPEI‐800 (1×10−3 mg/kg) provided significant neuroprotection. Consistent with the in vitro findings, the in vivo results following tMCAO also demonstrated a 100‐fold increase in the potency of the covalently linked compound UPEI‐800 compared to APPC alone.