| 名稱 | SP600125 |
| 描述 | SP600125 (JNK Inhibitor II) is a JNK inhibitor that inhibits JNK1, JNK2, and JNK3 (IC50=40/40/90 nM) with oral potency, reversibility, and ATP-competitive properties. SP600125 inhibits autophagy and induces apoptosis. |
| 細(xì)胞實(shí)驗(yàn) | Multiarray plate screening of mRNA was performed by High Throughput Genomics. In brief, cell lysates were prepared by using a single-step proprietary lysis buffer. Lysates were incubated with a 16-gene capture array manufactured into each well of a 96-well plate. Detection was by luminescence and was performed by HTG. SDs for triplicate samples were typically 3–8% for samples with high levels of gene expression and 15–25% for samples with very low (near-threshold) levels of cytokine gene expression [1]. |
| 動(dòng)物實(shí)驗(yàn) | Mouse LPS/TNF assay was performed as follows: Female CD-1 mice (8–10 weeks of age) were dosed i.v. or per os with SP600125 in PPCES vehicle (30% PEG-400/20% polypropylene glycol/15% Cremophor EL/5% ethanol/30% saline), final volume of 5 ml/kg, 15 min before i.v. injection with LPS in saline (0.5 mg/kg; Escherichia coli 055:B5). At 90 min, a terminal bleed was obtained from the abdominal vena cava, and the serum was recovered. Samples were analyzed for mouse TNF-α by using an ELISA. The in-life phase of the thymocyte apoptosis assay was performed in female C57BL/6 mice. SP600125 was administered at 0, 12, 24, and 36 h, 15 mg/kg s.c. in PPCES vehicle. Anti-CD3 (50 μg) i.p. (clone 145-2C11) was administered as a single dose immediately after SP600125 at time 0. After 48 h, mice were killed, and the thymus was dissected for thymocyte isolation. Treated and untreated mice thymuses were excised and immediately placed in complete medium (RPMI medium 1640 with 10% FBS, penicillin/streptomycin, and l-glutamine) on ice. Each thymus was then pressed between the frosted ends of 2 microscope slides to form a single cell suspension and collected through a 30 μm nylon mesh. Cells were stained for cell surface CD4 and CD8 and apoptosis and measured by flow cytometry [1]. |
| 體外活性 | 方法：小鼠肺癌細(xì)胞 LLC 和小鼠腫瘤細(xì)胞 4T1 用 SP600125 (3-10 μM) 處理 72 h，使用 MTT assay 檢測(cè)細(xì)胞活力。
結(jié)果：SP600125 劑量依賴性抑制 LLC 和 4T1 細(xì)胞生長(zhǎng)，IC50 為 8.14 μM 和 7.37 μM。[1]
方法：Jurkat T 細(xì)胞 用 SP600125 (1-50 μM) 預(yù)處理 10 min，隨后用 PMA (50 ng/mL)、anti-CD3 (0.5 μg/mL) 和 anti-CD28 (2 μg/mL) 刺激 30 min，使用 Western Blot 方法檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：SP600125 以 5-10 μM 的 IC50 阻斷 c-Jun 的磷酸化。在 50 μM 的濃度下，SP600125 不阻斷 ERK 的磷酸化，也不抑制 IκBα 的降解。在 50 μM 時(shí)觀察到 磷酸-p38 和 ATF2 的部分抑制，但在 25 μM 時(shí)沒有觀察到。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)對(duì) TNF-α 的抑制活性，將 SP600125 (7.5-30 mg/kg，30% PEG-400/20% polypropylene glycol/15% Cremophor EL/5% ethanol/30% saline) 靜脈注射或口服給藥給 CD-1 小鼠，15 min 后注射 LPS 誘導(dǎo)的 TNF-α 表達(dá)。
結(jié)果：靜脈注射 15 或 30 mg/kg SP600125 可顯著抑制 TNF-α 血清水平，而口服給藥劑量依賴性阻斷 TNF-α 表達(dá)，每次口服 30 mg/kg 可觀察到顯著抑制作用。[2]
方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 SP600125 (5 mg/kg) 和 C-2 (10 mg/kg) 腹腔注射給攜帶膀胱癌腫瘤 BIU87 的 nude 小鼠，每天一次，持續(xù)二十一天。
結(jié)果：C-2 治療抑制了腫瘤的生長(zhǎng)，C-2/SP600125 組腫瘤顯著低于用載體或 C-2 單獨(dú)治療的小鼠。[3] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Methanol : 1.25 mg/mL (5.68 mM), Sonication is recommended.
DMSO : 50 mg/mL (227.04 mM)
Ethanol : 1.1 mg/mL (5 mM)), Heating is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 2.2 mg/mL (9.99 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.
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| 關(guān)鍵字 | ATP-competitive | JNK | inhibit | Inhibitor | Apoptosis | SP-600125 | Ferroptosis | phosphorylation | SP 600125 | Autophagy | SP600125 | reversible |
| 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 |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 抗肝癌化合物庫 | 神經(jīng)退行性疾病化合物庫 | 疼痛相關(guān)化合物庫 | 酪氨酸激酶分子庫 |