Department of Neurology, Cedars-Sinai Medical Center, Burns and Allen Research Institute, 110 N. George Burns Road, D-2091, Los Angeles, CA 90048, USA.
Transl Stroke Res. 2011 Mar;2(1):51-9. doi: 10.1007/s12975-010-0034-4.
In the present study, we used a comprehensive cellular toxicity (CeeTox) analysis panel to determine the toxicity profile for CNB-001 [4-((1E)-2-(5-(4-hydroxy-3-methoxystyryl-)-1-phenyl-1H-pyrazoyl-3-yl)vinyl)-2-methoxy-phenol)], which is a hybrid molecule created by combining cyclohexyl bisphenol A, a molecule with neurotrophic activity and curcumin, a spice with neuro-protective activity. CNB-001 is a lead development compound since we have recently shown that CNB-001 has significant preclinical efficacy both in vitro and in vivo. In this study, we compared the CeeTox profile of CNB-001 with two neuroprotective molecules that have been clinically tested for efficacy: the hydrophilic free radical spin trap agent NXY-059 and the hydrophobic free radical scavenger edaravone (Radicut). CeeTox analyses using a rat hepatoma cell line (H4IIE) resulted in estimated C(Tox) value (i.e., sustained concentration expected to produce toxicity in a rat 14-day repeat dose study) of 42 μM for CNB-001 compared with >300 μM for both NXY-059 and Radicut. The CeeTox panel suggests that CNB-001 produces some adverse effects on cellular adenosine triphosphate content, membrane toxicity, glutathione content, and cell mass (or number), but only with high concentrations of the drug. After a 24-h exposure, the drug concentration that produced a half-maximal response (TC(50)) on the measures noted above ranges from 55 to 193 μM. Moreover, all CNB-001-induced changes in the markers were coincident with loss of cell number, prior to acute cell death as measured by membrane integrity, suggesting a cytostatic effect of CNB-001. NXY-059 and Radicut did not have acute toxic effects on H4IIE cells. We also found that CNB-001 resulted in an inhibition of ethoxyresorufin-o-deethylase activity, indicating that the drug may affect cytochrome P4501A activity and that CNB-001 was metabolically unstable using a rat microsome assay system. For CNB-001, an estimated in vitro efficacy/toxicity ratio is 183-643-fold, suggesting that there is a significant therapeutic safety window for CNB-001 and that it should be further developed as a novel neuroprotective agent to treat stroke.
在本研究中，我们使用了全面的细胞毒性（CeeTox）分析面板来确定 CNB-001[4-((1E)-2-(5-(4-羟基-3-甲氧基二苯乙烯基)-1-苯基-1H-吡唑基-3-基)乙烯基)-2-甲氧基苯酚]的毒性特征，这是一种由环己基双酚 A（具有神经营养活性的分子）和姜黄素（具有神经保护活性的香料）组合而成的混合分子。CNB-001 是一种先导化合物，因为我们最近发现 CNB-001 在体外和体内都具有显著的临床前疗效。在这项研究中，我们将 CNB-001 的 CeeTox 特征与两种已在临床中进行疗效测试的神经保护分子进行了比较：亲水性自由基捕获剂 NXY-059 和疏水性自由基清除剂依达拉奉（Radicut）。使用大鼠肝癌细胞系（H4IIE）进行的 CeeTox 分析得出，CNB-001 的估计 C(Tox) 值（即预计在大鼠 14 天重复剂量研究中产生毒性的持续浓度）为 42 μM，而 NXY-059 和 Radicut 的估计 C(Tox) 值均>300 μM。CeeTox 分析表明，CNB-001 对细胞三磷酸腺苷含量、膜毒性、谷胱甘肽含量和细胞质量（或数量）产生一些不良影响，但仅在药物高浓度时才会产生。在 24 小时暴露后，上述测量指标产生半最大反应（TC(50)）的药物浓度范围为 55 至 193 μM。此外，CNB-001 引起的所有标记物变化都与细胞膜完整性测量的急性细胞死亡之前的细胞数量减少同时发生，表明 CNB-001 具有细胞抑制作用。NXY-059 和 Radicut 对 H4IIE 细胞没有急性毒性作用。我们还发现 CNB-001 抑制了乙氧基-resorufin-o-脱乙基酶活性，表明该药物可能影响细胞色素 P4501A 活性，并且 CNB-001 在大鼠微粒体测定系统中代谢不稳定。对于 CNB-001，估计的体外疗效/毒性比为 183-643 倍，这表明 CNB-001 具有显著的治疗安全性窗口，应该进一步开发为治疗中风的新型神经保护剂。
