p53 (IC50 = 0.37 μM); NF-κB (IC50 = 0.47 μM); FACT

CBL0137 是胰腺癌细胞系细胞凋亡的有效诱导剂，对胰腺癌干细胞和活跃增殖的肿瘤细胞有毒。使用 CBL0137 和相关分子可以抑制 NF-B 和 HSF-1 控制的细胞应激途径，同时激活 p53 [2]。 CBL0137 结合 DNA 时不会引入任何类型的化学改变，使其无基因毒性。促进染色质转录 (FACT) 复合物是一种参与转录、复制和 DNA 修复的染色质重塑复合物，由于 CBL0137 与 DNA 结合而功能失活。在 CBL0137 处理的细胞中，FACT 从核质中丢失并被捕获在染色质中，从而抑制 FACT 依赖性转录，包括 NF-kB 介导的转录。此外，FACT 的染色质捕获导致 p53 以酪蛋白激酶 2 (CK2) 依赖性方式磷酸化和激活 [3]。

在小鼠中，CBL0137 可有效对抗多种胰腺导管腺癌 (PDA) 模型，包括原位吉西他滨耐药 PANC-1 模型和患者来源的异种移植物，其中 CBL0137 的抗肿瘤作用与 FACT 的过度表达相关[1]。根据其提出的作用机制，CBL0137靶向胶质母细胞瘤（GBM），穿透血脑屏障，并且在TMZ反应性和耐药性原位模型中均有效。这种药物能够穿过血脑屏障，尤其是静脉注射时，其治疗中枢神经系统肿瘤的潜力令人鼓舞。在原位模型中，静脉注射药物比口服药物具有更高的生物利用度，因为它们在肿瘤组织中积累更多。 CBL0137 在脑组织中的正常积累不会导致可检测到的神经毒性[3]。

将 CBL0137 盐酸盐应用于 MiaPaca2 和 BxPC-3 细胞 4 或 24 小时。蛋白酶和磷酸酶抑制剂存在于 1× 细胞培养裂解试剂中，用于收获细胞。在 SDS-PAGE 凝胶上分离后，将 5 至 20 μg 裂解物转移到 PVDF 膜上。靶向 SSRP1、SPT16、RRM1 和 RRM2 蛋白的抗体用于探测印迹。上样对照是 GAPDH。 ECL 试剂盒用于可视化蛋白质[1]。

将细胞重悬于无血清 Dulbecco's Modified Eagle Medium (DMEM) 中，并暴露于不同浓度的 CBL0137 盐酸盐中 1 小时。然后，将来自每种处理条件的 105 个细胞接种到 6 孔板的 3 个孔中，加入 2 mL 无血清 DMEM/F12 培养基，补充有 0.4% BSA、0.2×B27、10 ng/mL 重组 EGF，并含有 0.25 ％琼脂糖。在具有常规含 FBS 培养基的 6 孔板的三个孔中，将来自每种处理条件的 103 个细胞铺板。平板接种后七至十五天，在倒置显微镜下对菌落进行计数。

Ketamine/xylazine is used to create a deep anesthesia in 10-week-old female athymic nude mice (n = 8 per treatment group). Each mouse has its pancreas tail inoculated with 2 106 PANC-1 cells via laparotomy. When the tumor was detected by ultrasound two weeks after the vaccination, treatment started. There are the following routines: 1) Vehicles, gavage of sterile water and 100 mg/kg captisol 2) 100 mg/mL captisol diluted with 50 to 90 mg/kg CBL0137 hydrochloride and given intravenously once a week via the tail vein; 3) 10 to 20 mg/kg CBL0137 hydrochloride given orally five days on and two days off. To measure tumors, digital calipers are used. The equation LW2/2 is used to determine the tumor volume, where L is the longest dimension and W is the dimension that is perpendicular to W. Mice are monitored for at least 90 days after the start of treatment or until at least one tumor per mouse reached 1000 mm3, whichever occurs first.

[1]. Sci Transl Med . 2011 Aug 10;3(95):95ra74.

[2]. Oncotarget . 2014 Nov 30;5(22):11038-53.

[3]. Neuro Oncol . 2017 Feb 1;19(2):186-196.

Effective eradication of cancer requires treatment directed against multiple targets. The p53 and nuclear factor κB (NF-κB) pathways are dysregulated in nearly all tumors, making them attractive targets for therapeutic activation and inhibition, respectively. We have isolated and structurally optimized small molecules, curaxins, that simultaneously activate p53 and inhibit NF-κB without causing detectable genotoxicity. Curaxins demonstrated anticancer activity against all tested human tumor xenografts grown in mice. We report here that the effects of curaxins on p53 and NF-κB, as well as their toxicity to cancer cells, result from "chromatin trapping" of the FACT (facilitates chromatin transcription) complex. This FACT inaccessibility leads to phosphorylation of the p53 Ser(392) by casein kinase 2 and inhibition of NF-κB-dependent transcription, which requires FACT activity at the elongation stage. These results identify FACT as a prospective anticancer target enabling simultaneous modulation of several pathways frequently dysregulated in cancer without induction of DNA damage. Curaxins have the potential to be developed into effective and safe anticancer drugs.[1]

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Pancreatic ductal adenocarcinoma (PDA) continues to be one of the deadliest cancers due to the absence of effective treatment. Curaxins are a class of small molecules with anti-cancer activity demonstrated in different models of cancer in mice. The lead curaxin compound, CBL0137, recently entered Phase I clinical trials. Curaxins modulate several important signaling pathways involved in the pathogenesis of PDA through inhibition of chromatin remodeling complex FACT. FACT is overexpressed in multiple types of tumor, with one of the highest rate of overexpression in PDA (59%). In this study, the efficacy of CBL0137 alone or in combination with current standard of care, gemcitabine, was tested against different models of PDA in vitro and in mouse models. It was found that CBL0137 alone is a potent inducer of apoptosis in pancreatic cancer cell lines and is toxic not only for proliferating bulk tumor cells, but also for pancreatic cancer stem cells. In mice, CBL0137 was effective against several PDA models, including orthotopic gemcitabine resistant PANC-1 model and patient derived xenografts, in which CBL0137 anti-tumor effect correlated with overexpression of FACT. Moreover, we observed synergy of CBL0137 with gemcitabine which may be explained by the ability of CBL0137 to inhibit several transcriptional programs induced by gemcitabine, including NF-kappaB response and expression of ribonucleotide reductase, one of the targets of gemcitabine in cells. This data suggest testing of CBL0137 efficacy in Phase II trial in PDA patients alone and in combination with gemcitabine.[2]

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Pancreatic ductal adenocarcinoma (PDA) continues to be one of the deadliest cancers due to the absence of effective treatment. Curaxins are a class of small molecules with anti-cancer activity demonstrated in different models of cancer in mice. The lead curaxin compound, CBL0137, recently entered Phase I clinical trials. Curaxins modulate several important signaling pathways involved in the pathogenesis of PDA through inhibition of chromatin remodeling complex FACT. FACT is overexpressed in multiple types of tumor, with one of the highest rate of overexpression in PDA (59%). In this study, the efficacy of CBL0137 alone or in combination with current standard of care, gemcitabine, was tested against different models of PDA in vitro and in mouse models. It was found that CBL0137 alone is a potent inducer of apoptosis in pancreatic cancer cell lines and is toxic not only for proliferating bulk tumor cells, but also for pancreatic cancer stem cells. In mice, CBL0137 was effective against several PDA models, including orthotopic gemcitabine resistant PANC-1 model and patient derived xenografts, in which CBL0137 anti-tumor effect correlated with overexpression of FACT. Moreover, we observed synergy of CBL0137 with gemcitabine which may be explained by the ability of CBL0137 to inhibit several transcriptional programs induced by gemcitabine, including NF-kappaB response and expression of ribonucleotide reductase, one of the targets of gemcitabine in cells. This data suggest testing of CBL0137 efficacy in Phase II trial in PDA patients alone and in combination with gemcitabine.[3]

C21H24N2O2-HCL

372.89

372.16

C, 67.64; H, 6.76; Cl, 9.51; N, 7.51; O, 8.58

1197397-89-9

CBL0137;1197996-80-7

44519123

Off-white to yellow solid powder

5.39

51.1

2

3

6

26

466

0

CC(C1C=C2C(N(CCNC(C)C)C3=CC=C(C=C32)C(C)=O)=CC=1)=O.Cl

IXRKBBVMDMKAEB-UHFFFAOYSA-N

InChI=1S/C21H24N2O2.ClH/c1-13(2)22-9-10-23-20-7-5-16(14(3)24)11-18(20)19-12-17(15(4)25)6-8-21(19)23;/h5-8,11-13,22H,9-10H2,1-4H3;1H

1-[6-acetyl-9-[2-(propan-2-ylamino)ethyl]carbazol-3-yl]ethanone;hydrochloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

配方 1 中的溶解度: ≥ 2.5 mg/mL (6.70 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (6.70 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: 5%DMSO+40%PEG300+5%Tween80+50%ddH2O: 0.75mg/ml

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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：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。