PF-3845 is a dual inhibitor targeting fatty acid amide hydrolase (FAAH) and soluble epoxide hydrolase (sEH), with EC50 values of 0.65nM and 12000nM, respectively[1]. PF-3845 covalently inactivates FAAH by carbamylation of the enzyme's serine nucleophile without reacting with other serine hydrolases in vivo[2]. PF-3845 acts as a novel chemical scaffold inhibiting phenylalanyl-tRNA synthetase in Mycobacterium tuberculosis[3].
In vitro, PF-3845 treatment for 48h significantly inhibited the viability of Colo-205 cells with an IC50 value of 51.38μM[4]. Treatment with 10μM PF-3845 for 4 days inhibited receptor activator of nuclear factor kappa B ligand (RANKL)-induced differentiation of mouse bone marrow stromal cells (BMM) into osteoclasts and significantly decreased the mRNA expression of dendritic cell-specific transmembrane protein and cathepsin K in cells[5]. Treatment with 10μM PF-3845 for 18 hours significantly inhibited lipopolysaccharide (LPS)-induced prostaglandin E2 (PGE2) production and reduced inflammatory gene expression in BV2 cells[6].
In vivo, PF-3845 treatment via intraperitoneal injection at a dose of 5mg/kg/day for 14 days significantly restored hippocampal-dependent memory ability and alleviated traumatic brain injury (TBI)-induced anxiety and fine motor deficits in mice with TBI[7]. A single intraperitoneal injection of PF-3845 (5mg/kg) for 60 minutes can improve the plasma corticosterone release and glucose mobilization in rats after acute restraint stress[8].
References:
[1] Kodani S D, Wan D, Wagner K M, et al. Design and potency of dual soluble epoxide hydrolase/fatty acid amide hydrolase inhibitors[J]. ACS omega, 2018, 3(10): 14076-14086.
[2] Ahn K, Johnson D S, Mileni M, et al. Discovery and characterization of a highly selective FAAH inhibitor that reduces inflammatory pain[J]. Chemistry & biology, 2009, 16(4): 411-420.
[3] Wang H, Xu M, Engelhart C A, et al. Rediscovery of PF-3845 as a new chemical scaffold inhibiting phenylalanyl-tRNA synthetase in Mycobacterium tuberculosis[J]. Journal of Biological Chemistry, 2021, 296.
[4] Wasilewski A, Krajewska U, Owczarek K, et al. Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study[J]. Acta Biochimica Polonica, 2017, 64(3).
[5] Ihn H J, Kim Y S, Lim S, et al. PF-3845, a fatty acid amide hydrolase inhibitor, directly suppresses osteoclastogenesis through ERK and NF-κB pathways in vitro and alveolar bone loss in vivo[J]. International Journal of Molecular Sciences, 2021, 22(4): 1915.
[6] Tanaka M, Yagyu K, Sackett S, et al. Anti-inflammatory effects by pharmacological inhibition or knockdown of fatty acid amide hydrolase in BV2 microglial cells[J]. Cells, 2019, 8(5): 491.
[7] Tchantchou F, Tucker L B, Fu A H, et al. The fatty acid amide hydrolase inhibitor PF-3845 promotes neuronal survival, attenuates inflammation and improves functional recovery in mice with traumatic brain injury[J]. Neuropharmacology, 2014, 85: 427-439.
[8] Chen H J C, Spiers J G, Sernia C, et al. Inhibition of fatty acid amide hydrolase by PF-3845 alleviates the nitrergic and proinflammatory response in rat hippocampus following acute stress[J]. International Journal of Neuropsychopharmacology, 2018, 21(8): 786-795.
PF-3845是一种双靶点抑制剂，可同时靶向脂肪酸酰胺水解酶（FAAH）和可溶性环氧化物水解酶（sEH），对FAAH和sEH的EC50值分别为0.65nM和12000nM[1]。PF-3845通过羧基化FAAH酶的丝氨酸亲核基团，实现对酶的特异性共价抑制，且在体内不与其他丝氨酸水解酶反应[2]。PF-3845可作为新型化学支架，抑制结核分枝杆菌的苯丙氨酰-tRNA合成酶[3]。
在体外，PF-3845处理48小时能显著抑制Colo-205细胞活力，IC50值为51.38μM[4]。用10μM的PF-3845处理小鼠骨髓基质细胞4天，可抑制RANKL诱导的破骨细胞分化，并显著降低细胞中树突状细胞特异性跨膜蛋白和组织蛋白酶K的mRNA表达[5]。用10μM的PF-3845预处理BV2细胞18小时，能显著抑制脂多糖诱导的前列腺素E2产生，并降低炎症基因表达[6]。
在体内，通过每日腹腔注射5mg/kg剂量的PF-3845，连续14天，可显著恢复创伤性脑损伤小鼠的海马依赖性记忆能力，并缓解小鼠焦虑症状和精细运动缺陷[7]。对大鼠单次腹腔注射5mg/kg剂量的PF-3845（60分钟），能够改善急性束缚应激引起的血浆皮质酮释放和葡萄糖动员[8]。