In our recent studies, we identified compound N-benzyl-2-(2,5-dioxopyrrolidin-1-yl)propanamide (AS-1) as a broad-spectrum hybrid anticonvulsant which showed potent protection across the most important animal acute seizure models such as the maximal electroshock (MES) test, the subcutaneous pentylenetetrazole (s.c. PTZ) test, and the 6-Hz (32 mA) test in mice. Therefore, AS-1 may be recognized as a candidate for new anticonvulsant effective in different types of human epilepsy with a favorable safety margin profile determined in the rotarod test in mice. In the aim of further pharmacological evaluation of AS-1, in the current study, we examined its activity in the 6-Hz (44 mA) test, which is known as the model of drug-resistant epilepsy. Furthermore, we determined also the antiseizure activity in the kindling model of epilepsy induced by repeated injection of pentylenetetrazole (PTZ) in mice. As a result, AS-1 revealed relatively potent protection in the 6-Hz (44 mA) test, as well as delayed the progression of kindling induced by repeated injection of PTZ in mice at doses of 15 mg/kg, 30 mg/kg, and 60 mg/kg. Importantly, the isobolographic analysis showed that a combination of AS-1 and valproic acid (VPA) at the fixed ratio of 1:1 displayed a supra-additive (synergistic) interaction against PTZ-induced seizures in mice. Thus, AS-1 may be potentially used in an add-on therapy with VPA. Moreover, incubation of zebrafish larvae with AS-1 substantially decreased the number, cumulative but not the mean duration of epileptiform-like events in electroencephalographic assay. Finally, the in vitro ADME-Tox studies revealed that AS-1 is characterized by a very good permeability in the parallel artificial membrane permeability assay test, excellent metabolic stability on human liver microsomes (HLMs), no significant influence on CYP3A4/CYP2D6 activity, and moderate inhibition of CYP2C9 in a concentration of 10 μM, as well as no hepatotoxic properties in HepG2 cells (concentration of 10 μM).

中文翻译：

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具有混合结构的 N-苄基-(2,5-二氧吡咯烷-1-基)丙酰胺 (AS-1) 作为广谱抗癫痫药物的候选药物。


在我们最近的研究中，我们发现化合物N -benzyl-2-(2,5-dioxopyrrolidin-1-yl)propanamide (AS-1) 是一种广谱混合抗惊厥药，对最重要的动物急性癫痫发作模型显示出有效的保护作用例如最大电击（MES）测试、皮下戊四唑（sc PTZ）测试和小鼠6-Hz（32 mA）测试。因此，AS-1可能被认为是对不同类型的人类癫痫症有效的新型抗惊厥药物的候选者，并且在小鼠转棒试验中确定了良好的安全裕度。为了进一步评价AS-1的药理学，在本研究中，我们在6Hz（44mA）测试中检查了其活性，该测试被称为耐药性癫痫模型。此外，我们还测定了小鼠重复注射戊四唑（PTZ）诱导的癫痫点燃模型的抗癫痫活性。结果，AS-1在6-Hz（44 mA）测试中显示出相对有效的保护作用，并延迟了小鼠重复注射15 mg/kg、30 mg/kg剂量的PTZ引起的点燃进程。和 60 毫克/千克。重要的是，等辐射分析表明，AS-1 和丙戊酸 (VPA) 以 1:1 的固定比例组合，对 PTZ 诱导的小鼠癫痫发作表现出超加成（协同）相互作用。因此，AS-1 可能与 VPA 一起用于附加疗法。此外，将斑马鱼幼虫与AS-1一起孵育显着降低了脑电图检测中癫痫样事件的累积数量，但并未降低其平均持续时间。 最后，体外ADME-Tox研究表明，AS-1在平行人工膜渗透性测定试验中具有非常好的渗透性，在人肝微粒体（HLMs）上具有优异的代谢稳定性，对CYP3A4/CYP2D6活性无显着影响，浓度为 10 μM 时对 CYP2C9 具有中度抑制作用，并且对 HepG2 细胞（浓度为 10 μM）没有肝毒性。