Lenalidomide (CC-5013)是一种TNF-α分泌抑制剂，在PBMCs中IC50为13 nM。

靶点

体外研究

CC-5013强诱导IL-2和sIL-2R产量。CC-5013作用于T细胞，诱导CD28的酪氨酸磷酸化，随后下调NF-κB的激活。Lenalidomide和Pomalidomide作用于表达与Thalidomide结合野生型CRBN的，而不是Thalidomide结合缺陷型CRBN(YW/AA)的HEK293 T细胞，抑制CRBN自体泛素化。KMS12多发性骨髓瘤细胞中，CRBN野生型蛋白而不是CRBN(YW/AA) 突变型蛋白的过表达, 放大Pomalidomide调节的c-myc和IRF4表达降低和p21(WAF-1) 表达增高。H929 多发性骨髓瘤细胞系中长期抗Lenalidomide 的选择性与CRBN降低相关,然而抗Pomalidomide和 Lenalidomide的DF15R多发性骨髓瘤 中, 检测不到CRBN蛋白。Lenalidomide通过下调肿瘤细胞抑制性分子表达而抑制缺陷发生。Lenalidomide防止肿瘤诱导的T细胞裂解功能障碍发生。Lenalidomide 作用于T细胞，抑制慢性淋巴细胞性白血病（CLL）细胞诱导的T细胞肌动蛋白突触功能受损, 且下调CLL抑制配体及其他受体的表达。Lenalidomide 作用于FL, DLBCL, HL, MM, SCC,和 OC，抑制肿瘤诱导的免疫抑制，且作用于所有检测的肿瘤细胞时,下调免疫抑制配体表达

体内研究

口服处理Lenalidomide，显著抑制bFGF诱导的血管生成，这种作用存在剂量依赖性。Lenalidomide 显著降低血管面积百分数，从对照组的5.16%降低到实验组的2.58（50 mg/kg剂量处理）和 1.69（250 mg/kg 剂量处理）。 Lenalidomide 显著降低全部MVL，从对照组的21.07降低到实验组的8.11（50 mg/kg剂量处理）和1.90（250 mg/kg剂量处理）。

1 mg

5 mg

10 mg

1 mM

3.8571 mL

19.2857 mL

38.5713 mL

5 mM

0.7714 mL

3.8571 mL

7.7143 mL

10 mM

0.3857 mL

1.9286 mL

3.8571 mL

50 mM

0.0771 mL

0.3857 mL

0.7714 mL

Assay for inhibition of TNF synthesis by human PBMCs:
Human PBMCs from normal donors are obtained by Ficoll−Hypaque density centrifugation. Cells (106 cells/mL) are cultured in RPMI supplemented with 10 AB+ serum, 2 mM l-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin. Lenalidomide is dissolved in DMSO at 20 mg/mL; further dilution is done with culture medium. The final DMSO concentration in all assays including the controls is 0.25%. Lenalidomide is added to cells 1 hour prior to the addition of LPS. PBMCs (106 cells/mL) are stimulated with 1 μg/mL of LPS from Salmonella minnesota R595. Cells, in triplicate, are incubated with LPS for 18−20 hours at 37 °C in 5% CO2. Supernatants are then harvested and assayed for cytokine levels. In some experiments, supernatants are kept frozen at −70 °C until use. Cell viability is assayed by Trypan blue exclusion dye method. The concentration of TNFα in the culture supernatants is determined by ELISA. Lenalidomide is assayed in a minimum of three separate experiments. Percent inhibition is determined as 100 × [1 − (cytokine(experimental)/cytokine(control))].

细胞实验

• Cell lines: 良性或恶性 B (CLL)细胞, RPMI8226 MM 细胞
• Concentrations: 1 μM或5 nM
• Incubation Time: 24小时
• Method: 使用CellTracker Blue CMAC对良性或恶性B (CLL)细胞(2 × 106)进行染色， 然后使用2 μg/mL金黄色葡萄球菌超抗原(sAg) (SEA 和 SEB) 在37oC下进行脉冲处理30分钟。B细胞和等量T细胞(从原代共培养中纯化的)按200 × g转速离心5分钟，然后与CD8+ T 细胞在37oC下温育10分钟，或者与 CD3+ T细胞温育20分钟。使用细胞收集器使细胞转移到显微镜载玻片上，然后在室温下和3% 溶于PBS的甲醛混合15分钟。在完全培养基中加入终浓度为1 μM的Lenalidomide。对照组则使用DMSO处理细胞。

动物实验

• Animal Models: 携带HUVECs细胞的成年雄性Sprague–Dawley大鼠
• Formulation: 0.5% DMSO
• Dosages: 50 mg/kg和250 mg/kg
• Administration: 腹腔注射

TESTS

SPECIFICATIONS

Appearance

White-like powder

Solubility

30mg/ml DMSD,clear

UV

λmax=306±1nm; λmix=274±1nm

Identification

HNMR

Purity(HPLC)

> 99.0%