| 名稱(chēng) | JNJ-7706621 |
| 描述 | JNJ-7706621 is a potent aurora kinase inhibitor, and also inhibits CDK1 and CDK2. |
| 細(xì)胞實(shí)驗(yàn) | Cell lines: HeLa,HCT-116,A375,SK-OV-3,MDA-MB-231 and PC-3 cells. Concentrations: 1 nM - 10 μM,dissolved in DMSO. Method: Measuring incorporation of 14C-labelled thymidine into newly synthesized DNA within the cells to determine the ability of JNJ-7706621 to inhibit the proliferation of cell growth.Cells are trypsinized and counted and 3-8 ×103 cells are added to each well of a 96-well CytoStar tissue culture treated scintillating microplate in 100 μL complete medium in a volume.Cells are incubated for 24 hours at 37 °C in an atmosphere containing 5% CO2.Next,1 μL JNJ-7706621 is added to the wells of the plate.Cells are incubated for another 24 hours.Methyl 14C-thymidine 56 mCi/mmol is diluted in complete medium and 0.2 μCi/well is added to each well of the CytoStar plate in a volume of 20 μL.The plate is incubated for 24 hours at 37 °C in JNJ-7706621 with 14C-thymidine.The contents of the plate are discarded and the plate is washed twice with 200 μL PBS.200 μL of PBS is added to each well.The top of the plate is sealed with a transparent plate sealer and a white plate backing sealer is applied to the bottom of the plate.The degree of methyl 14C-thymidine incorporation is quantified on a Packard Top Count. |
| 激酶實(shí)驗(yàn) | In vitro kinase assay for CDK1 and Aurora kinases:CDK1 kinase activity is tested by the CDK1/cyclin B complex purified from baculovirus to phosphorylate a biotinylated peptide substrate containing the consensus phosphorylation site for histone H1, which is phosphorylated in vivo by CDK1. Inhibition of CDK1 activity is measured by observing a decreased amount of 33P-γ-ATP incorporation into the immobilized substrate in streptavidin-coated 96-well scintillating microplates. CDK1 enzyme is diluted in 50 mM Tris-HCl (pH 8), 10 mM MgCl2, 0.1 mM Na3VO4, 1 mM DTT, 1% DMSO, 0.25 μM peptide, 0.1 μCi per well 33P-γ-ATP, and 5 μM ATP in the presence or absence of various concentrations of JNJ-7706621 and incubated at 30 °C for 1 hour. The reaction is terminated by washing with PBS containing 100 mM EDTA and plates are counted in a scintillation counter. IC50 is determined by Linear regression analysis of the percent inhibition by JNJ-7706621.The Aurora kinase activity is measured with 10 μM ATP and a peptide containing a dual repeat of the kemptide phosphorylation motif. |
| 動(dòng)物實(shí)驗(yàn) | Animal Models: Mouse xenograft model of A375 cellsFormulation & . Dosages: Dissolved in 0.5% methylcellulose containing 0.1% polysorbate 80 in sterile water.100 or 125 mg/kg. Administration: Orally or by intraperitoneal injection |
| 體外活性 | JNJ-7706621（100或125 mg/kg）導(dǎo)致攜帶A375惡性黑色素瘤人類(lèi)腫瘤移植瘤模型的小鼠中腫瘤消退. |
| 體內(nèi)活性 | JNJ-7706621在抑制正常細(xì)胞類(lèi)型（包括MRC-5,HASMC,HUVEC和HMVEC）的生長(zhǎng)方面具有幾倍的效力,IC50為3.67-5.42 μM。JNJ-7706621對(duì)CDK1和2具有高度有效性,IC50為3-9 nM。JNJ-7706621顯示對(duì)包括HeLa,HCT-116,SK-OV-3,PC3,DU145,A375,MDA-MB-231,MES-SA和MES-SA/Dx5在內(nèi)的一組人類(lèi)癌細(xì)胞類(lèi)型的抑制作用 ,IC50為112-514 nM,獨(dú)立于p53,成視網(wǎng)膜細(xì)胞瘤或P-糖蛋白狀態(tài)。JNJ-7706621也抑制CDK3,4和6,IC50為58-253 nM。JNJ-7706621抑制Aurora-A和B,IC50分別為11和15 nM。JNJ-7706621也抑制VEGF-R2,FGF-R2和GSK3β,IC50為154-254 nM。在HeLa或U937細(xì)胞中,JNJ-7706621（0.5-3 μM）延遲從G1期中退出,使細(xì)胞停滯在G2-M期,誘導(dǎo)核內(nèi)復(fù)制,激活凋亡并減少集落形成。 |
| 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 : 55 mg/mL (139.47 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | Cyclin dependent kinase | CDK | Apoptosis | JNJ7706621 | inhibit | Inhibitor | Aurora Kinase | JNJ-7706621 |
| 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) | 抗癌活性化合物庫(kù) | 抗癌化合物庫(kù) | 已知活性化合物庫(kù) | 細(xì)胞周期化合物庫(kù) | 抗胰腺癌化合物庫(kù) | 激酶抑制劑庫(kù) | 含氟化合物庫(kù) | 抗衰老化合物庫(kù) |