Carvalho Gustavo Almeida, Chiareli Raphaela Almeida, Marques Bruno Lemes, Parreira Ricardo Cambraia, de Souza Gil Eric, de Carvalho Flávio Silva, da Rocha André Luís Batista, Silva Rafaela Ribeiro, Noël François, Vaz Boniek Gontijo, Lião Luciano Morais, Ahmad Shabir, Verli Hugo, Menegatti Ricardo, Pinto Mauro Cunha Xavier
Department of Pharmacology, Federal University of Goias, Goiânia, GO, Brazil.
Faculty of Pharmacy, Federal University of Goias, Goiânia, GO, Brazil.
Pharmacol Rep. 2023 Apr;75(2):276-292. doi: 10.1007/s43440-023-00451-x. Epub 2023 Jan 31.
L-proline transporter (PROT/SLC6A7) is closely associated with glutamatergic neurotransmission, where L-proline modulates the NMDA receptor (NMDAR) function. NMDAR-mediated excitotoxicity is a primary cause of neuronal death following stroke, which is triggered by the uncontrolled release of glutamate during the ischemic process. After ischemic stroke, L-proline levels show a reduction in the plasma, but high circulating levels of this molecule indicate good functional recovery. This work aimed to produce new PROT inhibitors and explore their effects on ischemic stroke.
Initially, we built a three-dimensional model of the PROT protein and run a molecular docking with the newly designed compounds (LQFM215, LQFM216, and LQFM217). Then, we synthesized new PROT inhibitors by molecular hybridization, and proline uptake was measured in ex vivo and in vivo models. The behavioral characterization of the treated mice was performed by the open-field test, elevated plus-maze, Y-maze, and forced swimming test. We used the permanent middle cerebral artery occlusion (MCAO) model to study the ischemic stroke damage and analyzed the motor impairment with limb clasping or cylinder tests.
LQFM215 inhibited proline uptake in hippocampal synaptosomes, and the LQFM215 treatment reduced proline levels in the mouse hippocampus. LQFM215 reduced the locomotor and exploratory activity in mice and did not show any anxiety-related or working memory impairments. In the MCAO model, LQFM215 pre-treatment and treatment reduced the infarcted area and reduced motor impairments in the cylinder test and limb clasping.
This dataset suggests that the new compounds inhibit cerebral L-proline uptake and that LQFM215 promotes neuroprotection and neuro-repair in the acute ischemic stroke model.
L-脯氨酸转运体（PROT/SLC6A7）与谷氨酸能神经传递密切相关，其中L-脯氨酸调节N-甲基-D-天冬氨酸受体（NMDAR）功能。NMDAR介导的兴奋性毒性是中风后神经元死亡的主要原因，其由缺血过程中谷氨酸的失控释放引发。缺血性中风后，血浆中L-脯氨酸水平降低，但该分子的高循环水平表明功能恢复良好。本研究旨在制备新的PROT抑制剂并探索其对缺血性中风的影响。
首先，我们构建了PROT蛋白的三维模型，并与新设计的化合物（LQFM215、LQFM216和LQFM217）进行分子对接。然后，通过分子杂交合成新的PROT抑制剂，并在体外和体内模型中测量脯氨酸摄取。通过旷场试验、高架十字迷宫、Y迷宫和强迫游泳试验对治疗小鼠进行行为表征。我们使用永久性大脑中动脉闭塞（MCAO）模型研究缺血性中风损伤，并通过肢体紧握或圆筒试验分析运动障碍。
LQFM215抑制海马突触体中的脯氨酸摄取，并且LQFM215处理降低了小鼠海马中的脯氨酸水平。LQFM215降低了小鼠的运动和探索活动，并且未显示出任何焦虑相关或工作记忆损伤。在MCAO模型中，LQFM215预处理和处理减少了梗死面积，并减少了圆筒试验和肢体紧握中的运动障碍。
该数据集表明新化合物抑制脑内L-脯氨酸摄取，并且LQFM215在急性缺血性中风模型中促进神经保护和神经修复。
