| 名稱 | Ro-3306 |
| 描述 | Ro 3306 is a CDK1 inhibitor that inhibits CDK1, CDK1/cyclin B1, and CDK2/cyclin E (Ki=20/35/340 nM) selectively and ATP-competitively. Ro-3306 exhibits antitumor activity, inhibits cell cycle arrest, and induces apoptosis. |
| 細(xì)胞實(shí)驗(yàn) | Log phases cells (25,000) are seeed in 96-well plates and incubated in a 37℃ incubator with CO2, After 24 h, different concentrations of RO-3306 are administered to determine the drug concentrations required to achieve a 50% growth inhibition (IC50). MTT (20 μL, 5 mg/mL stock solution in saline) is added to each well and the cells are incubated for 4 h. Supernatants are removed and formazan crystals from viable cells are solubilized with 200 μL anhydrous DMSO. The absorbance is detected with a 550 model microplate reader at the 565 nm wavelength.(Only for Reference) |
| 激酶實(shí)驗(yàn) | CDK assay: The activity of CDK1 cyclin B1, CDK1 cyclin A, CDK2 cyclin E, and CDK4 cyclin D is measured by a homogeneous time-resolved fluorescence assay in a 96-well format. The assay buffer contained 25 mM Hepes, 6.25 mM MgCl2, 0.003% Tween 20, 0.3 mg/mL BSA, 1.5 mM DTT, and ATP as follows: 162 μM (CDK1), 90 mM (CDK2), or 135 μM (CDK4). CDK1 and CDK2 buffer contained 10 mM MgCl2. Test compounds are diluted in assay buffer to 3-fold their final concentration in 20 μL, and the reaction is started by the addition of a 40 μL assay buffer containing the pRB substrate (0.185 μM). The plates are incubated at 37°C for 30 min with constant agitation, and the reaction is terminated by the addition of 15 μL of 1.6 μM anti-phospho pRB antibody (Ser-780) in 25 mM Hepes, 24 mM EDTA, and 0.2 mg/mL BSA. After an additional 30 min of incubation with shaking, 15μL of 3 nM Lance-Eu-W1024-labeledanti-rabbitIgG and 60 nM Alophycocyanin-conjugated anti-His-6 antibody in 25 mM Hepes, and 0.5 mg/mL BSA is added and incubated for 1 h. The plates are read in the Victor-V multi- label reader at excitation 340 nm and emission 615 nm and 665 nm. The IC50 values are calculated from the readings at 665 nm and normalized for Europium readings at 615 nm. Ki values are calculated according to the equation: Ki= IC50/(1 + S/Km ), where S is the ATP concentration in the assay and Km is the Michaelis-Menten constant for ATP. The inhibitory activity against the panel of kinases is determined by the IMAP assay technology. |
| 體外活性 | 方法：卵巢癌細(xì)胞 OVCA-429 和 OVCAR-3 用 Ro 3306 (1-2.5 nM) 處理 9 天，使用 crystal violet assay 檢測(cè)細(xì)胞活力。
結(jié)果：OVCA-429 和 OVCAR-3 細(xì)胞在高劑量濃度的 Ro 3306 下長(zhǎng)達(dá) 9 天的生長(zhǎng)速率較低。使用 2.5 μM Ro 3306 處理，OVCA-429 和 OVCAR-3 細(xì)胞的生長(zhǎng)率在第 9 天分別降低了 75.3% 和 87.7%。[1]
方法：人腫瘤細(xì)胞系 HCT116、SW480 和 HeLa 用 Ro 3306 (9 μM) 處理 20 h，使用 Flow cytometry 檢測(cè)細(xì)胞周期。
結(jié)果：用 Ro 3306 對(duì)增殖的人類腫瘤細(xì)胞處理 20 h 導(dǎo)致 G2/M 期細(xì)胞周期的完全阻斷。[2] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Ro 3306 (4 mg/kg) 和 Cisplatin (4 mg/kg) 腹腔注射給攜帶卵巢癌腫瘤 OVCAR-3 的 BALB/c nude 小鼠，每四天一次，持續(xù)四周。
結(jié)果：與 Ro 3306 或 Cisplatin 治療相比，聯(lián)合治療有效抑制了腫瘤生長(zhǎng)。[1] |
| 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 | DMSO : 12.5 mg/mL (35.57 mM)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 1.25 mg/mL (3.56 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 | Cyclin dependent kinase | Ro3306 | Inhibitor | Apoptosis | CDK | Ro 3306 | inhibit | Ro-3306 |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | Sorafenib | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | 抗胰腺癌化合物庫(kù) | 激酶抑制劑庫(kù) | 抗衰老化合物庫(kù) | TGF-β/Smad靶點(diǎn)化合物庫(kù) | 糖代謝化合物庫(kù) | 酪氨酸激酶分子庫(kù) | 疼痛相關(guān)化合物庫(kù) |