Neuronal nitric oxide synthase (nNOS) is an enzyme constitutively expressed in the mammalian brain and skeletal muscles. The excessive activation of nNOS in the neurons results in oxidative and nitrosative stress associated with neuronal loss in various neurological disorders. Several nNOS inhibitors have been reported to limit the excessive activation of nNOS. In the present work, we have designed and carried the synthesis of benzo[d]thiazol-2-yl-methyl-4-(substituted)-piperazine-1-carbothioamide as novel neuronal nitric oxide inhibitors (5–28, twenty-four compounds). Stably transfected HEK 293 cells expressing NOS isoforms treated with the compounds (5–28) showed that the eight compounds exhibited > 95% cell survival in the MTT assay. nNOS inhibition assay of the eight compounds illustrated that the compound 18 was most selective for nNOS (nNOS=66.73 ± 1.51; eNOS=28.70 ± 1.39; iNOS =13.26 ± 1.01) in HEK 293 cells expressing NOS isoforms. 6-OHDA-induced unilaterally lesioned rats treated with the compound 18 showed the improvement in motor and non-motor functions. Furthermore, the compound 18 showed the increased levels of dopamine and decreased levels of glutamate and nitrite ions in the isolated rat brain. In the docking analysis, the compound 18 showed the significant binding affinity with the nNOS binding site (the ∆G value = - 9.0 kcal/mol). Overall results demonstrated that the N-(benzo[d]thiazol-2-ylmethyl)-4-(4-nitrophenyl) piperazine-1-carbothioamide (the compound 18) possessed significant nNOS inhibiting activity and neuroprotecting potential in 6-OHDA-induced unilaterally lesioned rat model of PD and more work will be required to establish the role of the compound 18 in the therapy of PD and other neurodegenerative disorders.

神经元一氧化氮合酶 (nNOS) 是一种在哺乳动物大脑和骨骼肌中组成性表达的酶。神经元中 nNOS 的过度激活导致与各种神经系统疾病中的神经元丢失相关的氧化和亚硝化应激。据报道，几种 nNOS 抑制剂可以限制 nNOS 的过度激活。在目前的工作中，我们设计并合成了苯并[ d ]噻唑-2-基-甲基-4-（取代的）-哌嗪-1-硫代甲酰胺作为新型神经元一氧化氮抑制剂（5-28，二十四化合物）。表达经化合物处理的 NOS 亚型的稳定转染 HEK 293 细胞 ( 5–28)显示八种化合物在 MTT 测定中表现出 > 95% 的细胞存活率。八种化合物的 nNOS 抑制测定表明化合物18在表达 NOS 亚型的 HEK 293 细胞中对 nNOS 最具选择性（nNOS = 66.73 ± 1.51；eNOS = 28.70 ± 1.39；iNOS = 13.26 ± 1.01）。用化合物18治疗的 6-OHDA 诱导的单侧损伤大鼠表现出运动和非运动功能的改善。此外，化合物18显示离体大鼠大脑中多巴胺水平升高，谷氨酸和亚硝酸根离子水平降低。在对接分析中，化合物18显示与 nNOS 结合位点的显着结合亲和力（ΔG 值 = - 9.0 kcal/mol）。总体结果表明，N-(benzo[ d ]thiazol-2-ylmethyl)-4-(4-nitrophenyl) piperazine-1-carbothioamide（化合物18）在 6-OHDA 诱导的中具有显着的 nNOS 抑制活性和神经保护潜力PD 的单侧损伤大鼠模型，需要做更多的工作来确定化合物18在治疗 PD 和其他神经退行性疾病中的作用。