CRBN; TNF-α (IC50 = 13 nM)

（放大倍数，×200）。Pomalidomide 抑制人 PBMC 和人全血中脂多糖 (LPS) 刺激的 TNF-α 释放的 IC50 分别为 13 nM 和 25 nM。 [1] Pomalidomide 的 IC50 为 1 μM，抑制 IL-2 诱导的 T 调节细胞生长。 [2] 泊马度胺 (6.4 nM–10 M) 治疗会导致人外周血 T 细胞中 IL-2 的产生增加；这种效应在 CD4+ 子集中比在 CD8+ 子集中稍微更明显。 Pomalidomide 比 CC-5013 具有更强的增加 IL-2、IL-5 和 IL-10 水平的能力，但其增加 IFN-γ 水平的能力仅稍强一些。 Pomalidomide 以剂量依赖性方式增强 SEE 和 Raji 细胞在 Jurkat 细胞中诱导的 AP-1 转录活性，在 1 μM 时最大增强 4 倍。 [3] 当Raji细胞暴露于不同浓度的泊马度胺（2.5-40 μg/mL）48小时时，细胞增殖和DNA合成显着减少。与使用媒介物处理的对照相比，减少了 40%。 [4]

在患有严重联合免疫缺陷的小鼠中，泊马度胺可提高利妥昔单抗治疗 B 细胞淋巴瘤的能力。与CC5013/利妥昔单抗治疗的58天和利妥昔单抗非治疗的45天相比，泊马度胺和利妥昔单抗联合治疗的小鼠的中位生存期为74天。泊马度胺和利妥昔单抗具有协同作用，但这种作用可以通过 NK 细胞耗竭而完全逆转，这支持了以下观点：泊马度胺可能增加利妥昔单抗抗肿瘤活性的一种方式是促进 NK 细胞扩增。 [4]

在脂多糖 (LPS) 刺激的 PBMC 中测量 TNF-α 抑制活性。在添加 LPS (1 μg/mL) 之前一小时，将泊马度胺添加到人 PBMC 中，然后再继续孵育 18 至 20 小时。收获上清液后，使用 ELISA 测量上清液中 TNF-α 的浓度。使用非线性回归分析来确定将 TNF 产量减少 50% 所需的泊马度胺 (IC50) 量。与PBMC测定类似，进行人全血TNF抑制测定，不同之处在于将已肝素化的新鲜人全血直接铺在微量滴定板上。

将泊马度胺 (5 μg/mL) 应用于淋巴瘤细胞系 24 或 48 小时，以测量细胞凋亡。使用碘化丙啶和 FITC 标记的膜联蛋白 V 对细胞进行染色。荧光激活细胞分选仪/FACStar Plus流式细胞仪多色流式细胞分析用于检查细胞凋亡。当细胞表现出早期或晚期凋亡的迹象（分别为膜联蛋白 V 阳性和碘丙啶阴性或阳性）时，它们被认为是凋亡的。将淋巴瘤细胞系暴露于 Pomalidomide（2.5、5、10、20 和 40 μg/mL）24 或 48 小时以测量细胞增殖。每孔添加 1 μCi 的 [3H]-胸苷（在 96 孔板中）后，细胞再孵育 18 小时。使用 Harvest 系统收获细胞并将其放入 96 孔玻璃过滤器后，使用自动闪烁计数器测定 [3H]-胸苷摄取。

Mice: SCID mice aged six to eight weeks are used for this. All of the animals are injected with 1×106 Raji cells through their tail veins on day 0. The animals are divided into seven cohorts following 72 hours of tumor engraftment. The first cohort (group A) serves as the control and is not given any medication. Animals in Groups B and C were given either CC-5013 (0.5 mg/kg) or Pomalidomide (0.5 mg/kg) intravenously on Days +3, +4, +8, +9, +13, +14, +18, and +19. Rituximab or Trastuzumab (isotype control) monotherapy is administered to Groups D and E on Days +5, +10, +15, and +20 by tail vein injection at a dose of 10 mg/kg. Animals treated with Rituximab and CC-5013 (group E) or Pomalidomide (group G) make up groups F and G, respectively. Prior to each dose of Rituximab, IMiDs are administered intravenously for two consecutive days. Animals are monitored for 90 days after therapy is over. The study's primary outcome is survival, which is measured as the amount of time before limb paralysis sets in. Cervical dislocation is used to kill any animals that reach the end point or remain alive after three months of observation. To find any remaining disease, a pathologic examination of all organs is conducted, including the liver, lungs, and brain. Three different times, the experiments are repeated.
Rats: Three male CD-IGS rats in total are used. Pomalidomide is given as a single PO administration through the stomach cannula at a dose of 50 mg/kg (5 mL/kg) in a suspension formulation of 0.5% carboxymethylcellulose/0.25% Tween 80. Ten hours after dosing, microdialysate is collected in a cooling fraction collector set to 4°C at intervals of 25 minutes. Each sample's corrected concentration is multiplied by the sampling interval, in this case 25 minutes, and divided by the number of hours in a day to obtain the AUC. These values added together represented the overall AUC value for the given time frame. The concentration is plotted at each time point at the halfway point of each collection interval in order to create graphs. Within 12 hours of the specified time points, microdialysates are collected and analyzed for the presence of pomalidomide using a LC-MS/MS assay.

[1]. Molecular mechanism of action of the immune-modulatory drugs, thalidomide, lenalidomide and pomalidomide in multiple myeloma. Leuk Lymphoma. 2013 Apr;54(4):683-7.

[2]. Immunomodulatory drug CC-5013 or CC-4047 and rituximab enhance antitumor activity in a severe combined immunodeficient mouse lymphoma model. Clin Cancer Res. 2005 Aug 15;11(16):5984-92.

[3]. Enhancement of cytokine production and AP-1 transcriptional activity in T cells by thalidomide-related immunomodulatory drugs. J Pharmacol Exp Ther. 2003 Jun;305(3):1222-32.

[4]. Pomalidomide shows significant therapeutic activity against CNS lymphoma with a major impact on the tumor microenvironment in murine models. PLoS One. 2013 Aug 5;8(8):e71754.

[5]. Hijacking the E3 Ubiquitin Ligase Cereblon to Efficiently Target BRD4. Chem Biol. 2015 Jun 18;22(6):755-63.

[6]. Tumour necrosis factor-α inhibits hepatic lipid deposition through GSK-3β/β-catenin signaling in juvenile turbot (Scophthalmus maximus L.). Gen Comp Endocrinol. 2016 Mar 1;228:1-8.

C13H11N3O4

273.24

273.07496

C, 57.14; H, 4.06; N, 15.38; O, 23.42

19171-19-8

Pomalidomide-d3;2093128-28-8;Pomalidomide-d5;1377838-49-7;Pomalidomide-d4;1416575-78-4

white solid powder

C1CC(=O)NC(=O)C1N2C(=O)C3=C(C2=O)C(=CC=C3)N

UVSMNLNDYGZFPF-UHFFFAOYSA-N

InChI=1S/C13H11N3O4/c14-7-3-1-2-6-10(7)13(20)16(12(6)19)8-4-5-9(17)15-11(8)18/h1-3,8H,4-5,14H2,(H,15,17,18)

4-amino-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

CC4047; CC-4047; CC 4047; Pomalidomide. Brand name: Pomalyst

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

1%DMSO+30% polyethylene glycol+1%Tween 80: 15mg/mL

请根据您的实验动物和给药方式选择适当的溶解配方/方案：
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10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
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