与FDA批准的药物替莫唑胺相比,Carmofur是一种酸性神经酰胺酶抑制剂,在抑制和杀死致命的侵袭性胶质母细胞瘤细胞系方面具有卓越的功效。除了脑肿瘤,carmofur还作为SARS-CoV-2主要蛋白酶(MPRO)的潜在铅抑制剂而受到关注。据报道,它还对许多其他癌症和非癌症疾病,包括急性肺损伤有效。痴呆症,帕金森病,儿童室管膜瘤,和Krabbe病等。Carmofur还具有抗真菌和抗微生物特性。因此,需要一种灵敏的生物分析方法,以支持进一步的体内药理学研究,临床前和临床研究。在这里,我们报告一个敏感的,和可靠的LC-MS/MS方法用于使用小鼠血浆定量生物分析卡莫氟。使用乙酸乙酯和2-丙醇(85:15)的液-液萃取(LLE)技术制备样品。在XBridgeBEHC18XP色谱柱上实现色谱分离(100毫米×3毫米,2.5µm),运行时间为八分钟。定量是在多反应监测(MRM)模式下进行的,在负电喷雾电离中,对于Carmofur,前体到产物的离子跃迁为m/z256.25→m/z129.01,对于5-氯尿嘧啶(IS),则为m/z145.53→m/z42.00。Carmofur在5-1,000ng的范围内显示出良好的线性。mL-1。该方法在特异性方面进行了验证,线性度结转,基体效应,回收效率,准确度,精度,稀释完整性,和稳定性。最后,该方法已成功用于腹腔内给药后小鼠血浆中的药代动力学研究。据我们所知,这是用于卡莫弗生物分析的LC-MS/MS方法的第一份报告。
Carmofur is an acid ceramidase inhibitor with superior efficacy in suppressing and killing fatally aggressive glioblastoma cell lines compared to the FDA-approved drug temozolomide. In addition to brain tumors,
carmofur also gained attention as a potential lead inhibitor of the main protease (MPRO) of SARS-CoV-2. It is also reported efficacious against numerous other cancers and non-cancerous diseases including acute lung injury, dementia, Parkinson\'s disease, childhood ependymoma, and Krabbe disease etc.
Carmofur also possesses antifungal and antimicrobial properties. Therefore, a sensitive bio-analytical method is needed in order to support further in vivo pharmacological investigation, pre-clinical and clinical studies. Herein, we report a sensitive, and reliable LC-MS/MS method for quantitative bioanalysis of
carmofur using mouse plasma. The samples were prepared employing liquid-liquid extraction (LLE) technique using ethyl acetate and 2-propanol (85:15). Chromatographic separation was achieved on an XBridge BEH C18 XP column (100 mm × 3 mm, 2.5 µm) with a runtime of eight minutes. Quantification was performed in multiple reaction monitoring (MRM) mode with precursor to product ion transition of m/z 256.25 → m/z 129.01 for carmofur and m/z 145.53 → m/z 42.00 for 5-chlorouracil (IS) in negative electrospray ionization.
Carmofur showed good linearity over the range of 5-1,000 ng.mL-1. The method was validated in terms of specificity, linearity, carry-over, matrix effect, recovery efficiency, accuracy, precision, dilution integrity, and stability. Finally, the method was successfully employed in a pharmacokinetic study in mouse plasma after intraperitoneal administration of the drug solution. To the best of our knowledge, this is the first report of an LC-MS/MS method for carmofur bioanalysis.