CID-2858522 is an effective and specific antigen receptor-mediated NF-κB activation inhibitor (IC50: 70 nM).

NF-κB:70 nM

CID-2858522 (化合物1) 在人体肝微粒体和NADPH生成系统存在下抑制睾丸激素羟化酶的活性（IC50：85 μM）[1]。在HEK293细胞系中，CID-2858522以浓度依赖方式抑制NF-κB报告基因的活性，IC50约为70 nM，最大抑制作用在0.25-0.5 μM达到。相反，在高达4 μM的浓度下，CID-2858522不抑制TNF诱导的NF-κB报告基因活性。CID-2858522还强效抑制PMA/Ionomycin诱导的NF-κB报告基因活性，在瞬时转染试验中表现出来[2]。

CID-2858522表现出非线性药代动力学特性，在0.5小时测量时间点，30 mg/kg剂量的血清水平高于50 mg/kg剂量，但在t=3小时时表现出典型的剂量依赖性行为。50 mg/kg剂量时观察到的累积增加可能是由于CYP3A4抑制产生的库效应。在t=3小时，50 mg/kg剂量的队列明显显示出病理性征象[2]。

Cell viability is estimated based on cellular ATP levels. HEK293 cells at a density of 10^5/mL are seeded at 90 μL per well in 96-well white plates and cultured overnight. Compounds (e.g., CID-2858522; 1 μM, 2 μM, 3 μM, and 4 μM) are added (5 μL in medium) to wells and cells are cultured for 16 h, Finally, 50 μL ATPlite solution is added to each well and luminescence activity is read using a luminometer [2].

Three male mice are subjected to CID-2858522 (single i.p doses at 10, 30, and 50 mg/kg). Blood is drawn at 0.5 and 3 h, and subsequent LC/MS analysis of pooled samples is performed to determine the overall blood levels of CID-2858522 [2].