Low Yi Ling, Kreutzer Ethan, Chandrashekaran Indu R, Adams Luke A, Pun Jason, Doak Bradley C, Pan Yijun, Short Jennifer L, Scanlon Martin J, Nicolazzo Joseph A
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia.
Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, 3052, Australia.
J Neuroimmune Pharmacol. 2025 Apr 16;20(1):40. doi: 10.1007/s11481-025-10191-9.
Fatty acid-binding protein 4 (FABP4) is a key lipid binding protein expressed in microglia, which has been demonstrated to play a critical role in microglial-mediated neuroinflammation, a component of many neurodegenerative diseases. Compounds able to inhibit the function of FABP4 have shown promise in reducing microglial-mediated neuroinflammation, however, their physicochemical properties would prevent their ability to be easily formulated and traverse the blood-brain barrier (BBB) in order to access microglial FABP4. To this end, this study assessed the ability of a series of FABP4 inhibitors, with more desirable physicochemical properties, to attenuate microglial inflammation in an in vitro setting. Four inhibitors with varying affinity to FABP4, as measured by isothermal titration calorimetry (MFP-0011462, MFP-0012314, MFP-0012318, and MFP-0012328), were assessed for their ability to induce toxicity and attenuate reactive oxygen species (ROS) generation and tumour necrosis factor-α (TNF-α) release from lipopolysaccharide (LPS)-activated BV-2 microglia. All FABP4 inhibitors were determined to be soluble in the aqueous buffers at the highest concentration used in the assays (100 µM). Isothermal titration calorimetry demonstrated that the compounds had varying affinities for FABP4 (K values of 316 nM to > 100 µM). The ability of FABP4 inhibitors to reduce LPS-mediated ROS production aligned with their K for FABP4, with the most effective inhibitor (MFP-0012328) also able to reduce TNF-α production (by RT-qPCR) and TNF-α release from LPS-activated BV-2 cells by 17% and 25%, respectively. These studies have demonstrated that a series of FABP4 inhibitors with more appropriate physicochemical properties for BBB penetration are able to reduce microglial-mediated inflammation, which may be of benefit in diseases where overactivation of microglia leads to neurodegeneration.
脂肪酸结合蛋白4(FABP4)是一种在小胶质细胞中表达的关键脂质结合蛋白,已证明其在小胶质细胞介导的神经炎症中起关键作用,而神经炎症是许多神经退行性疾病的一个组成部分。能够抑制FABP4功能的化合物在减轻小胶质细胞介导的神经炎症方面显示出前景,然而,它们的物理化学性质会妨碍其易于制剂化并穿越血脑屏障(BBB)以作用于小胶质细胞FABP4的能力。为此,本研究评估了一系列具有更理想物理化学性质的FABP4抑制剂在体外环境中减轻小胶质细胞炎症的能力。通过等温滴定量热法测定了四种对FABP4具有不同亲和力的抑制剂(MFP-0011462、MFP-0012314、MFP-0012318和MFP-0012328)诱导毒性以及减轻脂多糖(LPS)激活的BV-2小胶质细胞中活性氧(ROS)生成和肿瘤坏死因子-α(TNF-α)释放的能力。所有FABP4抑制剂在测定中使用的最高浓度(100µM)下均能溶解于水性缓冲液中。等温滴定量热法表明这些化合物对FABP4具有不同的亲和力(K值为316 nM至>100µM)。FABP4抑制剂减少LPS介导的ROS产生的能力与其对FABP4的K值一致,最有效的抑制剂(MFP-0012328)还能够分别将LPS激活的BV-2细胞中的TNF-α产生(通过RT-qPCR)和TNF-α释放减少17%和25%。这些研究表明,一系列具有更适合BBB穿透的物理化学性质的FABP4抑制剂能够减轻小胶质细胞介导的炎症,这可能对小胶质细胞过度激活导致神经退行性变的疾病有益。