| 名稱 | Mirdametinib |
| 描述 | Mirdametinib (PD325901) is an MEK inhibitor (IC50=0.33 nM) with selective, non-ATP-competitive, and oral activity. Mirdametinib exhibits antitumor activity by inhibiting p-ERK1/2 expression and inducing apoptosis. |
| 細胞實驗 | A cell death detection enzyme-linked immunosorbent assay was used per the manufacturer's instructions. Briefly, 4 × 10^4 cells were plated in 24-well plates in triplicate the day before treatment. PTC cells were treated with 0.1 μmol/L PD0325901 for 96 hours. Cells treated with 1 μmol/L staurosporine served as positive controls for apoptosis. At the end of treatment, cells were lysed using the lysis buffer provided in the kit for 30 minutes at room temperature and then centrifuged in 24-well plates. Lysates (20 μL of supernatant) were transferred to streptavidin-coated wells and incubated for 2 hours at room temperature with two antibodies (biotin-labeled anti-histone antibody and peroxidase-conjugated anti-DNA antibody). After the wells were washed three times, the samples were incubated with peroxidase substrate (ABTS) and the amount of colored product was determined spectrophotometrically at 405 nm. The background was measured at 490 nm [2]. |
| 激酶實驗 | Incorporation of 32P into myelin basic protein (MBP) is assayed in the presence of a glutathione S-transferase fusion protein containing p44MAP kinase (GST-MAPK) and a glutathione S-transferase protein containing p45MEK (GST-MEK). The assay solution contained 20 mM HEPES, pH 7.4, 10 mM MgCl2, 1 mM MnCl2, 1 mM EGTA, 50 mM [γ-32P]ATP, 10 mg GST-MEK, 0.5 mg GST-MAPK and 40 mg MBP in a final volume of 100 mL. Reactions are stopped after 20 minutes by addition of trichloroacetic acid and filtered through a GF/C filter mat. 32P retained on the filter mat is determined using a 1205 Betaplate [1]. |
| 動物實驗 | Athymic Ncr-nu/nu mice were obtained from the National Cancer Institute at ages 6 to 8 weeks and housed for at least 1 week after arrival. Mice (10–14 per group) were anesthetized s.c. with a cocktail (100 μL/10 g body weight of 10 mg/mL ketamine and 1 mg/mL xylazine). K2 and TPC-1 cells stably infected with a retrovirus expressing luciferase (5 × 105 cells in 5 μL RPMI1640 medium) were inoculated into the thyroid gland, and the mice were monitored weekly for tumor growth by Xenogen (IVIS 200 imaging system) using Living Image 3.0 software. One week after inoculation, PD0325901 was dissolved in 80 mmol/L citric buffer (pH 7) by sonication and given to mice daily by oral gavage (20–25 mg/kg) for 3 weeks (5 consecutive days/week). Mice were sacrificed only due to tumor burden or loss of 20% of body weight. Tumor sizes were measured with calipers and tumor volume (V) was calculated by the formula (V = length × width × depth). Control mice were given 80 mmol/L citric buffer (pH 7) alone. All in vivo experiments were done at least twice [2]. |
| 體外活性 | 方法：11 種人黑色素瘤細胞系上用 Mirdametinib (0.1-1000 nM) 處理 72 h，使用 trypan blue exclusion test 檢測細胞數(shù)。
結(jié)果：Mirdametinib (IC50=20-50 nM) 有效抑制具有 BRAF 突變 (M14/A375P/A375M/A375SM/ME10538/ME4686/JR8) 或不具有 BRAF 突變 (ME4405/ME13923) 的人黑色素瘤細胞系的生長。ME1007 和 ME8959均具有野生型 BRAF，對 Mirdametinib介導(dǎo)的生長抑制的抗性略高 (IC50≥100 nM)。[1]
方法：乳頭狀甲狀腺癌 (PTC) 細胞系 K2 和 TPC-1 用 Mirdametinib (0.1-1000 nmol/L) 處理 1-96 h，使用 Western Blot 檢測靶點蛋白表達水平。
結(jié)果：Mirdametinib 可以有效抑制多種 PTC 細胞系中 ERK1/2 的磷酸化。[2] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)抗腫瘤活性，將 Mirdametinib (20-25 mg/kg，80 mmol/L citric buffer (pH 7)) 灌胃給藥給攜帶 PTC 腫瘤 K2 或 TPC-1 的 Athymic Ncr-nu/nu 小鼠，每周五次，持續(xù)三周。
結(jié)果：Mirdametinib 在接種攜帶 BRAF 突變的 PTC 細胞 K2 的小鼠中完全抑制腫瘤生長，并且在接種攜帶 RET/PTC1 重排的 PTC 細胞 TPC-1 的小鼠中顯著降低腫瘤生長。[2]
方法：為檢測體內(nèi)抗腫瘤活性，將 Mirdametinib (1.6-25 mg/kg，0.5% hydroxypropylmethylcellulose+0.2% Tween 80 in water) 口服給藥給攜帶小鼠結(jié)直腸癌腫瘤 CT26 的小鼠，每天一次，持續(xù)十四天。
結(jié)果：Mirdametinib 顯著抑制腫瘤中的 pERK 水平。[3] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | 10% DMSO+40% PEG300+5% Tween 80+45% Saline : 5 mg/mL (10.37 mM), Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
H2O : Insoluble
DMSO : 50 mg/mL (103.69 mM)
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| 關(guān)鍵字 | PD 0325901 | MAPKK | Mitogen-activated protein kinase kinase | Apoptosis | MEK | PD-0325901 | Autophagy | inhibit | MAP2K | PD-325901 | Inhibitor | PD 325901 | Mirdametinib |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | L-Glutamic acid | Gefitinib | Hydroxychloroquine | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | L-Ascorbic acid | Paeonol | Sodium 4-phenylbutyrate |
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