The cytolytic protein perforin has a crucial role in infections and tumor surveillance. Recently, it has also been associated with many brain diseases, such as neurodegenerative diseases and stroke. Therefore, inhibitors of perforin have attracted interest as novel drug candidates. We have previously reported that converting a perforin inhibitor into an L-type amino acid transporter 1 (LAT1)-utilizing prodrug can improve the compound’s brain drug delivery not only across the blood–brain barrier (BBB) but also into the brain parenchymal cells: neurons, astrocytes, and microglia. The present study evaluated whether the increased uptake into mouse primary cortical astrocytes and subsequently improvements in the cellular bioavailability of this brain-targeted perforin inhibitor prodrug could enhance its pharmacological effects, such as inhibition of production of caspase-3/-7, lipid peroxidation products and prostaglandin E₂ (PGE₂) in the lipopolysaccharide (LPS)-induced neuroinflammation mouse model. It was demonstrated that increased brain and cellular drug delivery could improve the ability of perforin inhibitors to elicit their pharmacological effects in the brain at nano- to picomolar levels. Furthermore, the prodrug displayed multifunctional properties since it also inhibited the activity of several key enzymes related to Alzheimer’s disease (AD), such as the β-site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1), acetylcholinesterase (AChE), and most probably also cyclooxygenases (COX) at micromolar concentrations. Therefore, this prodrug is a potential drug candidate for preventing Aβ-accumulation and ACh-depletion in addition to combatting neuroinflammation, oxidative stress, and neural apoptosis within the brain.

溶细胞蛋白穿孔素在感染和肿瘤监测中发挥着至关重要的作用。最近，它还与许多脑部疾病有关，例如神经退行性疾病和中风。因此，穿孔素抑制剂作为新型候选药物引起了人们的兴趣。我们之前曾报道，将穿孔素抑制剂转化为利用 L 型氨基酸转运蛋白 1 (LAT1) 的前药可以改善该化合物的脑药物递送，不仅可以穿过血脑屏障 (BBB)，还可以改善进入脑实质细胞的药物递送：神经元、星形胶质细胞和小胶质细胞。本研究评估了这种脑靶向穿孔素抑制剂前药对小鼠原代皮质星形胶质细胞的摄取增加以及随后细胞生物利用度的提高是否可以增强其药理作用，例如抑制 caspase-3/-7、脂质过氧化产物的产生和前列腺素E ₂ (PGE ₂ ) 在脂多糖(LPS) 诱导的神经炎症小鼠模型中的作用。研究表明，增加大脑和细胞药物输送可以提高穿孔素抑制剂在纳米至皮摩尔水平的大脑中引发药理作用的能力。此外，该前药表现出多功能特性，因为它还抑制与阿尔茨海默病（AD）相关的几种关键酶的活性，例如β位点淀粉样前体蛋白（APP）裂解酶1（BACE1）、乙酰胆碱酯酶（AChE）和最有可能的是微摩尔浓度的环氧合酶（COX）。因此，除了对抗大脑内的神经炎症、氧化应激和神经细胞凋亡之外，这种前药也是预防 Aβ 积累和 ACh 消耗的潜在候选药物。