| 名稱 | THZ1 |
| 描述 | THZ1 (CDK7 inhibitor) is a selective covalent inhibitor of CDK7, exhibiting binding affinity to the cysteine residue located at the outer end of the classical kinase domain, thus conferring high selectivity for CDK7, with an IC50 of 3.2 nM. |
| 細(xì)胞實(shí)驗(yàn) | Cells are treated with THZ1, THZ1-R or dimethylsulphoxide (DMSO) for 0-6?h to assess the effect of time on the THZ1-mediated inhibition of RNAPII CTD phosphorylation. For subsequent experiments cells are treated with compounds for 4?h as determined by the time-course experiment described earlier, unless otherwise noted. For inhibitor washout experiments, cells are treated with THZ1, THZ1-R or DMSO for 4?h. Medium containing inhibitors is subsequently removed to effectively 'washout' the compound and the cells are allowed to grow in the absence of inhibitor. For each experiment, lysates are probed for RNAPII CTD phosphorylation and other specified proteins.(Only for Reference) |
| 激酶實(shí)驗(yàn) | For kinase assays following immunoprecipitation of FLAG-CDK7 protein from HCT116 or FLAG-CDK12 from 293A cellular lysates, cells are first treated with THZ1, THZ1-R, or DMSO for 4 hrs at 37°C. Cells are then harvested by lysis in 50 mM Tris HCl pH 8.0, 150 mM NaCl, 1% NP-40, 5 mM EDTA, and protease/phosphatase cocktails. Exogenous CDK7 or CDK12 proteins are immunoprecipitated from cellular lysates using FLAG antibody- conjugated agarose beads. Precipitated proteins are washed with lysis buffer 6 times, followed by 2 washes with kinase buffer (40 mM Hepes pH 7.5, 150 mM NaCl, 10 mM MgCl2, 5% glycerol) and subjected to in vitro kinase assays at 30°C for 45 minutes using 1 μg of the large subunit of RNAPII (RPB1) as substrate and 25 μM ATP and 10 μCi of 32P ATP. Kinase assays using recombinant CDK7/TFIIH/MAT1 are conducted in the manner as described above using 25 ng of CAK complex per reaction. For kinase assays designed to test time-dependent inactivation of CDK7 kinase activity, CAK complex is pre-incubated with indicated concentrations of THZ1, THZ1-R, or DMSO in kinase buffer without ATP for 4 hrs at 30°C prior to being subjected to kinase assay conditions[1]. |
| 體外活性 | 方法：Loucy 細(xì)胞用 THZ1 (0.8-500 nM) 處理 4 h，通過 LanthaScreen Eu Kinase Binding assay 檢測結(jié)合情況。
結(jié)果：THZ1 在體外對 CDK7 具有時(shí)間依賴性抑制作用，并具有細(xì)胞內(nèi) CDK7 的共價(jià)結(jié)合作用。[1]
方法：NSCLC 細(xì)胞系 H1299、A549 和 H292 用 THZ1 (10-10000 nM) 處理 48 h，通過 crystal violet 檢測細(xì)胞活力。
結(jié)果：THZ1 劑量依賴性地抑制了 NSCLC 細(xì)胞的遷移增殖。[2] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)抗腫瘤活性，將 THZ1 (10 mg/kg) 靜脈注射給攜帶 MYCN 擴(kuò)增的人 NB 異種移植物的 NU/NU 小鼠，每天兩次，持續(xù) 28 天。
結(jié)果：THZ1 抑制人 MYCN 擴(kuò)增 NB 小鼠模型中的腫瘤生長。[3] 方法：為檢測體內(nèi)抗腫瘤活性，將 THZ1 (10 mg/kg) 腹腔注射給攜帶 KYSE510 腫瘤的 NSG 小鼠，每天兩次，持續(xù) 24 天。
結(jié)果：THZ1 抑制 KYSE510 異種移植物的生長并抑制 NSG 小鼠模型的肺轉(zhuǎn)移。[4] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 50 mg/mL (88.33 mM), Sonication is recommended.
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| 關(guān)鍵字 | THZ-1 | THZ1 | THZ 1 | CDK7 | CDK |
| 相關(guān)產(chǎn)品 | Ribociclib | Palbociclib monohydrochloride | Abemaciclib methanesulfonate | Abemaciclib | CASIN | Olomoucine | Palbociclib | GW 441756 | Dinaciclib | Ro-3306 | GSK 3 Inhibitor IX | SNS-032 |
| 相關(guān)庫 | 抑制劑庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 抗癌化合物庫 | 已知活性化合物庫 | 細(xì)胞周期化合物庫 | 抗胰腺癌化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 臨床前化合物庫 | 神經(jīng)退行性疾病化合物庫 | 共價(jià)抑制劑庫 |