In the present research, a lipophilic methotrexate (MTX) prodrug was developed by covalent conjugating of MTX to stearic acid (SA) as a lipid moiety via amid bond. The structure of synthesized conjugate, MTX-SA, was confirmed by IR and NMR spectra. To evaluate inflammatory response of MTX-SA conjugate and MTX, human PBMCs were isolated and exposed to 50, 500 and 5000 nM of MTX-SA conjugate and free MTX. The expression of four key genes involved in inflammation and apoptosis including IL-8, IL-1β, IL-10 and Bcl2 depicted that the MTX-SA had controversial behavior in different doses on the inflammatory transcription. Also, MTX-SA statically decreased the number of immune live cells in comparison to MTX. However, MTX-SA did not capture PBMCs cell cycle in G0/G1 phase. Totally, these results showed MTX-SA with long lipid chain has different effect on immune responses and it is irrefutable that detailed studies of its immunotoxicity and immunogenicity ought to be taken into account.

在本研究中，亲脂性甲氨蝶呤 (MTX) 前药是通过 MTX 与作为脂质部分的硬脂酸 (SA) 通过酰胺键共价结合而开发的。合成的共轭物 MTX-SA 的结构通过 IR 和 NMR 光谱得到证实。为了评估 MTX-SA 偶联物和 MTX 的炎症反应，将人 PBMC 分离并暴露于 50、500 和 5000 nM 的 MTX-SA 偶联物和游离 MTX。IL-8、IL-1β、IL-10 和 Bcl2 四个参与炎症和细胞凋亡的关键基因的表达表明 MTX-SA 在不同剂量下对炎症转录具有有争议的行为。此外，与 MTX 相比，MTX-SA 静态地减少了免疫活细胞的数量。然而，MTX-SA 没有捕获 G0/G1 期的 PBMCs 细胞周期。完全，