生物活性
VX-809 (Lumacaftor)通过促进突变型CFTR(F508del-CFTR)的成熟，从而纠正常见于囊性纤维化的CFTR突变，EC50为0.1 μM。VX-809在雌激素受体（ER）水平上起作用的行为，使一部分F508del-CFTR采取正确折叠的方式，退出ER，转移到细胞表面，正常起作用。VX-809作用于表达 F508del-CFTR的Fischer 大鼠甲状腺 (FRT)细胞, VX-809显著提高F508del-CFTR突变，提高7.1 倍，EC50为0.1 μM, 且增强F508del-CFTR调节的氯离子运输，提高5 倍， EC50 为0.5 μM, 而VRT-768作用时具有更高的EC50 值，EC50分别为7.9 μM 和 16 μM。VX-809 (3 μM)作用于表达F508del-CFTR的HEK-293细胞,提高ER中的F508del-CFTR，提高6倍。
化学数据
| 分子量 |
452.41 |
| 分子式 |
C24H18F2N2O5 |
| CAS号 |
936727-05-8 |
| 纯度 |
99.60% |
| 溶解性（25°C） |
DMSO 60 mg/mL |
| 储存和运输条件 |
固体粉末： -20°C 冷藏长期储存
常温运输及临时存放 |
实验操作来自于公开的文献，仅供相同实验参考（如实验材料、目的不同，请参考其他文献）
| 细胞实验 |
| 细胞系 |
FRT, HEK-293 and HBE cells |
| 方法 |
CFTR Immunoblot Analysis.
FRT, HEK-293, or HBE cells expressing CFTR or F508del-CFTR were incubated for 24 h at 37 °C with or without VX-809 in the assay media. After incubation, cells were harvested in ice-cold D-PBS solution (without calcium and magnesium) and pelleted at 1,000 × g at 4 °C. Cell pellets were lysed in 1% Nonidet P-40, 0.5% sodium deoxycholate, 200 mM NaCl, 10 mM Tris, pH 7.8, and 1 mM EDTA plus protease inhibitor mixture (1:250; Roche) for 30 min on ice. Lysates were spun for 10 min at 10,000 × g at 4 °C to pellet nuclei and insoluble material. Approximately 12 μg total protein was heated in Laemmli buffer with 5% β mercaptoethanol at 37 °C for 5 min and loaded onto a 3% to 8% Tris-acetate gel (Invitrogen). The gel was transferred to nitrocellulose and processed for Western blotting by using monoclonal CFTR antibody 769 (gift from John R. Riordan, University of North Carolina, Chapel Hill, NC) or polyclonal to GAPDH (Santa Cruz Biotechnology). Blots were developed by enhanced chemiluminescence. Quantification of the relative amounts of bands C and GAPDH was performed by using NIH ImageJ analysis of scanned films. |
| 浓度 |
0~100μM |
| 处理时间 |
48 h |
| 动物实验 |
| 动物模型 |
Male Sprague–Dawley rats model |
| 配制 |
0.5% Tween80/0.5% methylcellulose/water |
| 剂量 |
1 mg/kg |
| 给药处理 |
orally |
不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）
|
小鼠 |
大鼠 |
兔 |
豚鼠 |
仓鼠 |
狗 |
| 重量 (kg) |
0.02 |
0.15 |
1.8 |
0.4 |
0.08 |
10 |
| 体表面积 (m2) |
0.007 |
0.025 |
0.15 |
0.05 |
0.02 |
0.5 |
| Km系数 |
3 |
6 |
12 |
8 |
5 |
20 |
| 动物 A (mg/kg) = 动物 B (mg/kg) × |
动物 B的Km系数 |
| 动物 A的Km系数 |
例如，依据体表面积折算法，将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量，需要将20 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到化合物用于大鼠的等效剂量为10 mg/kg。
储备液配制
以下数据基于半岛bd体育手机客户端 分子量，对于特殊半岛bd体育手机客户端 ，请参照COA中的储备液配制条件和说明进行操作。
| Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
| 1 mM |
2.2104 mL |
11.0519 mL |
22.1038 mL |
| 5 mM |
0.4421 mL |
2.2104 mL |
4.4208 mL |
| 10 mM |
0.221 mL |
1.1052 mL |
2.2104 mL |
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