PD184352 (CI1040)   中文名称: PD-184352 (CI-1040)

MEK1 (IC50 = 17 nM); MEK2 (IC50 = 17 nM)

CI-1040 的 IC50 为 17 nM，直接抑制 MEK1。此外，一组 IC50 值高出 2.5 个数量级的相关激酶已被证明对其几乎没有活性。当用 CI-1040 处理整个细胞时，有丝分裂原诱导的 ERK 磷酸化被完全阻断。在 MDA-MB-231 乳腺癌细胞中，浓度为 1 μM 的 CI-1040 可分别抑制 99% 和 92% 的 ERK1 和 ERK2 磷酸化[1]。在 U-937 细胞中，CI-1040 以剂量和时间依赖性方式引起细胞凋亡并阻止增殖。 CI-1040 后 PUMA mRNA 和蛋白质水平显着升高[2]。

MEK 抑制剂 CI-1040 的全身给药显着改善了肺结构，同时将腺瘤的形成减少了三分之一。 CL-1040 治疗的小鼠肺细胞增殖较少，对肺细胞分化没有明显影响[3]。

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在体内，MEK抑制剂CI-1040的全身给药将腺瘤形成减少到第三个，并显著恢复了肺结构。CL-1040处理的小鼠肺细胞增殖率降低，对肺细胞分化没有明显影响。相比之下，Raf抑制剂BAY 43-9006不影响体内腺瘤的形成。 结论：MEK抑制剂CI-1040可用于治疗Ras和/或Raf依赖性人类恶性肿瘤。[3]

髓磷脂碱性蛋白 (MBP) 在被 MEK 激活后，会被激活的 MAP 激酶磷酸化。当谷胱甘肽 S-转移酶 (GST) 融合蛋白由 44-kDa MAPK (GST-MAPK) 或 45-kDa MEK (GST) 制成时-MEK1) 存在，测量 32P 掺入髓鞘碱性蛋白 (MBP) 的情况。测定在 50 mL 50 mM Tris、pH 7.4、10 mM MgCl2、2 mM EGTA 和 10 μM [γ-32P]ATP 中进行，其中含有 10 μg GST-MEK1、0.5 μg GST-MAPK 和 40 μg的MBP。 30°C 孵育 15 分钟后，添加 Laemmli SDS 样品缓冲液终止反应。 SDS/10% PAGE 可解析磷酸化的 MBP。该筛选过程的结果是鉴定出多种小分子 MEK 抑制剂，包括 CI-1040。 50% 抑制浓度 (IC50) 为 17 nM，检查添加顺序的实验表明 CI-1040 直接抑制 MEK1，而不影响 MAPK 的活性。

在细胞培养中，MEK 抑制剂 CI-1040 溶解在 DMSO 中制成 10 mM 储备液后，最终浓度为 50 mg/mL。用 5 和 20 uM CI-1040 预处理 24 小时后，用 wt-p53 siRNA 或 PUMA siRNA 转染 U-937 细胞 48 小时。然后每孔接受20mL MTT溶液，并且该过程再重复2小时。为了将代谢活细胞产生的MTT甲臜溶解在100mL异丙醇中，实验结束后吸出上清液。将板在旋转摇床上混合 30 分钟后，使用读板器测量 595 nm 处的吸光度[2]。

Mice: By administering constitutively active C-Raf kinase to the lung, a lung cancer mouse model is produced. A daily intraperitoneal injection of BAY 43-9006 or CI-1040 is given starting at the age of 4 months for a total of 21 days at a dose of 100 mg/kg. At the conclusion of the treatment period, the lungs were separated and examined[3].

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In this study, researchers have generated a lung cancer mouse model by targeting constitutively active C-Raf kinase to the lung. These mice develop adenomas within 4 months of life. At this time-point they received daily intraperitoneal injections of either 100 mg/kg BAY 43-9006 or CI-1040 for additional 21 days. Thereafter, lungs were isolated and the following parameters were analyzed using histology and immunohistochemistry: overall lung structure, frequency of adenoma foci, proliferation rate, ERK activity, caspase-3 activation, and lung differentiation.[3]

[1]. CI-1040 (PD184352), a targeted signal transduction inhibitor of MEK (MAPKK). Semin Oncol. 2003 Oct;30(5 Suppl 16):105-16.

[2]. MEK inhibitor CI-1040 induces apoptosis in acute myeloid leukemia cells in vitro. Eur Rev Med Pharmacol Sci. 2016 May;20(10):1961-8.

[3]. Use of mitogenic cascade blockers for treatment of C-Raf induced lung adenoma in vivo: CI-1040strongly reduces growth and improves lung structure. BMC Cancer. 2004 Jun 1;4:24.

2-(2-chloro-4-iodoanilino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide is an aminobenzoic acid.
CI-1040 has been used in trials studying the treatment of Lung Cancer, Breast Cancer, Breast Neoplasms, Pancreatic Cancer, and Colorectal Cancer, among others.
MEK Inhibitor CI-1040 is an agent that inhibits both mitogen-activated protein kinase kinases 1 and 2 (MEK1 and MEK2), substrates of Raf and phosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2), preventing phosphorylation and activation of the Mitogen-Activated Protein Kinase (MAPK) pathways, involved with signal transduction pathways and tumor proliferation.

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Several key growth factors, cytokines, and proto-oncogenes transduce their growth- and differentiation-promoting signals through the mitogen-activated protein kinase or extracellular signal-regulated protein kinase (ERK) cascade. Overexpression or constitutive activation of this pathway has been shown to play an important role in the pathogenesis and progression of breast and other cancers, making the components of this signaling cascade potentially important as therapeutic targets. CI-1040 (PD184352) is an orally active, highly specific, small-molecule inhibitor of one of the key components of this pathway (MEK1/MEK2), and thereby effectively blocks the phosphorylation of ERK and continued signal transduction through this pathway. Antitumor activity has been seen in preclinical models with this compound, particularly for pancreas, colon, and breast cancers, which has been shown to correlate with its inhibition of pERK. Clinically, CI-1040 has been shown to be well tolerated in phase I studies, with safety and pharmacokinetic profiles that permit continuous daily dosing. Biomarker studies have shown target inhibition in patients, and antitumor activity has also been observed with a partial response in one patient with pancreatic cancer and stable disease in approximately 25% of phase I patients. Given the central role of the ERK/mitogen-activated protein kinase pathway in mediating growth-promoting signals for a diverse group of upstream stimuli, inhibitors of MEK, as a key central mediator, could have significant clinical benefit in the treatment of breast and other cancers.[1]

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Objective: MEK1/2 (mitogen-activated protein kinase 1 and 2)/ERK1/2 (extracellular signal-regulated kinase 1 and 2) is important transducers of external signals for cell growth, survival, and apoptosis in acute myeloid leukemia cells (AML). In this study, we analyzed the effect of MEK inhibitor CI-1040 on the survival of AML cells. Materials and methods: Using ELISA and MTT we studied the cytotoxic effects of CI-1040 on AML U-937 cells. We studied the changes induced by CI-1040 on PUMA and p53 expression in U-937 cells by Western blotting assay. Moreover, we analyzed the cytotoxic effect of CI-1040 in U-937 cells with deleted PUMA, wt-p53 by wt-p53 siRNA and PUMA siRNA transfection. Results: CI-1040 induced apoptosis and inhibited proliferation in U-937 cells in a dose and time-dependent manner. CI-1040 induced a significant increase in PUMA mRNA and protein levels. Importantly, we show that knockdown of PUMA by PUMA siRNA transfection inhibited CI-1040-induced apoptosis and proliferation inhibition in U-937 cells. Moreover, CI-1040 induced apoptosis and proliferation inhibition was irrespective of wt-P53 status. Conclusions: These results demonstrate that CI-1040 induce apoptosis of U-937 cells and might be a new therapeutic option for the treatment of AML.[2]

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Background: Signaling networks promoting cell growth and proliferation are frequently deregulated in cancer. Tumors often are highly dependent on such signaling pathways and may become hypersensitive to downregulation of key components within these signaling cascades. The classical mitogenic cascade transmits stimuli from growth factor receptors via Ras, Raf, MEK and ERK to the cell nucleus and provides attractive molecular targets for cancer treatment. For example, Ras and Raf kinase inhibitors are already in a number of ongoing phase II and phase III clinical trials. In this study the effect of the Raf kinase inhibitor BAY 43-9006 and of the MEK inhibitor CI-1040 (PD184352) on a Raf dependent lung tumor mouse model was analyzed in detail. Methods: We have generated a lung cancer mouse model by targeting constitutively active C-Raf kinase to the lung. These mice develop adenomas within 4 months of life. At this time-point they received daily intraperitoneal injections of either 100 mg/kg BAY 43-9006 or CI-1040 for additional 21 days. Thereafter, lungs were isolated and the following parameters were analyzed using histology and immunohistochemistry: overall lung structure, frequency of adenoma foci, proliferation rate, ERK activity, caspase-3 activation, and lung differentiation. Results: Both inhibitors were equally effective in vitro using a sensitive Raf/MEK/ERK ELISA. In vivo, the systemic administration of the MEK inhibitor CI-1040 reduced adenoma formation to a third and significantly restored lung structure. The proliferation rate of lung cells of mice treated with CL-1040 was decreased without any obvious effects on differentiation of pneumocytes. In contrast, the Raf inhibitor BAY 43-9006 did not influence adenoma formation in vivo. Conclusion: The MEK inhibitor CI-1040 may be used for the treatment of Ras and/or Raf-dependent human malignancies.[3]

C17H14CLF2IN2O2

478.67

477.975

C, 42.66; H, 2.95; Cl, 7.41; F, 7.94; I, 26.51; N, 5.85; O, 6.69

212631-79-3

6918454

White to off-white solid powder

1.7±0.1 g/cm3

166-169ºC

1.656

8.01

50.36

2

5

6

25

472

0

IC1C([H])=C([H])C(=C(C=1[H])Cl)N([H])C1C(=C(C([H])=C([H])C=1C(N([H])OC([H])([H])C1([H])C([H])([H])C1([H])[H])=O)F)F

GFMMXOIFOQCCGU-UHFFFAOYSA-N

InChI=1S/C17H14ClF2IN2O2/c18-12-7-10(21)3-6-14(12)22-16-11(4-5-13(19)15(16)20)17(24)23-25-8-9-1-2-9/h3-7,9,22H,1-2,8H2,(H,23,24)

2-(2-chloro-4-iodoanilino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide

PD 184352; CI-1040; PD184352 (CI-1040); 2-(2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide; CI1040; PD-184352; 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide; PD184352 (CI-1040); 2-(2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide; PD184352; CI 1040

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.22 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: ≥ 2.08 mg/mL (4.35 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 30% PEG400+0.5% Tween80+5% propylene glycol, pH 9: 10mg/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。