I-BET-762 别名：GSK525762
I-BET-762是一种BET (bromodomain and extra terminal domain)蛋白抑制剂， IC50约为35 nM，抑制巨噬细胞产生促炎蛋白，抑制急性炎症，对其他溴区结合域包含的蛋白具有高度的选择性。
CAS No.：1260907-17-2
JCI Insight. 2022 Sep 8;7(17):e151851.
Targeting ESR1 mutation-induced transcriptional addiction in breast cancer with BET inhibition
化学性质/溶解性/储存
| 分子量 | 423.9 |
| 分子式 | C22H22ClN5O2 |
| CAS号 | 1260907-17-2 |
| 溶解性 | DMSO 90 mg/mL 1M HCl: 70 mg/mL |
| 储存条件 |
粉末型式 -20°C 3年；4°C 2年 溶于溶剂 -80°C 6个月；-20°C 1个月 |
| 运输方式 | 冰袋运输，根据产品的不同，可能会有相应调整。 |
生物活性
实验参考
下述溶液配置方法仅为基于分子量计算出的理论值。不同产品在配置溶液前，需考虑其在不同溶剂中的溶解度限制。
| 浓度/溶剂体积/质量 | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 1 mM | 2.359 mL | 11.7952 mL | 23.5905 mL |
| 5 mM | 0.4718 mL | 2.359 mL | 4.7181 mL |
| 10 mM | 0.2359 mL | 1.1795 mL | 2.359 mL |
*吸湿的DMSO对产品的溶解度有显著影响，请使用新开封的DMSO；
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
| 细胞系 | isolated CD4+ T cells |
| 方法 | CD4+ T cells are isolated from lymph nodes and spleens of 10- to 12-wk-old 2D2 transgenic mice by using Dynal Beads according to manufacturer’s protocols. The isolated CD4+ T cells were stimulated with platebound anti-CD3 and anti-CD28 antibodies alone (ThN conditions), or in the presence of anti-IL4 and IL-12 (Th1 conditions), IL-4 and anti-IL12 (Th2 conditions), IL-1β, IL-6, and IL-23 (Th17 conditions), or TGF-β (Treg conditions). Along with the cytokine mixtures were included either the Control-768 (GSK525768A) or I-BET-762 (GSK525762A) compounds. The cells are stimulated in these condition for 60–72 h and then harvested and expanded with DMEM media containing 10% (vol/vol) FBS, antibiotics, Lglutamine, and B-2ME. The cells were counted and reset to 0.5 × 10^6 cells per mL on days 3 and 4 after initial stimulation. Over the course of 5 d of T-cell culture and expansion, the compounds were diluted 12-fold relative to the starting concentrations. In case of Th1, Th2, and iTreg conditions, IL-2 was added to the media at a final concentration of 20 units per mL. On day 5 after initial activation, the cells were harvested and restimulated with PMA (10 nM) and ionomycin (1 μM) for 6 h. Brefeldin A (10 μg/mL) was added during the last 2 h of stimulation. Subsequently, the cells were fixed with 4% (wt/vol) paraformaldehyde in PBS for 15 min at 25 °C, washed in PBS, and permeabilized in saponin buffer [PBS, 0.5% saponin (AbMole), 1% (wt/vol) BSA, and 0.1% (wt/vol) sodium azide]. Intracellular staining was performed as described for indicated cytokines. In some experiments, cell surface staining was completed before fixation. |
| 浓度 | 125, 250, 500nM |
| 处理时间 | 60-72h |
* 上述方法来自公开文献，仅供相同目的实验参考。如实验目的、材料、方法不同，请参考其他文献。
建议您制定动物给药及实验方案时，尽量参考已发表的相关实验文献（溶剂种类及配比众多，简单地溶解目的化合物，并不能解决动物给药依从性、体内生物利用度、组织分布等相关问题，未必能保证目的化合物在动物体内充分发挥生物学效用）。
体内实验的工作液，建议您现用现配，当天使用；如在配制过程中出现沉淀、析出现象，可以通过超声和（或）加热的方式助溶。
切勿一次性将产品全部溶解。
请在下面的计算器中，输入您的动物实验相关数据并点击计算，即可得到该实验的总需药量和工作液终浓度。
例如您给药剂量是10 mg/kg，平均每只动物的体重为20 g，每只动物的给药体积是100 μL，动物数量为20只，则该动物实验的总需药量为4 mg，工作液终浓度为2 mg/mL。
1：鉴于实验过程的损耗，建议您至少多配1-2只动物的量；
2：为该产品最终给药时的浓度。
| 动物模型 | LPS-induced endotoxic shock of C57BL/6 mice model |
| 配制 | 20% beta-cyclodextrin, 2% DMSO in 0.9% saline |
| 剂量 | 30 mg/kg |
| 给药处理 | retro-orbital or tail vein injection |
* 上述方法来自公开文献，仅供相同目的实验参考。如实验目的、材料、方法不同，请参考其他文献。
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