Roscovitine (Seliciclib,CYC202)

Roscovitine (Seliciclib,CYC202)は有効な細胞週期卵白の依存性キナーゼ選択阻害剤、cdc2/ cyclin B、cdk2/ cyclin A、cdk2/ cyclin Eとcdk5/ p53に作用する時、IC50それぞれ0.65、0.7、0.7と0.16μM。

目録号S1153
5 5 4レビュー 8製品表彰状
価格 在庫  
USD 185 In stock
USD 240 In stock
USD 592 In stock
USD 1222 In stock

Roscovitine (Seliciclib,CYC202) 化学構造
分子量: 354.45

品質と確認

カスタマーレビュー(4)

Quality Control & MSDS

製品情報

  • Compare CDK Inhibitors
    CDK阻害剤を比較
  • 研究分野
  • Roscovitine (Seliciclib,CYC202)のメカニズム

製品の説明

生物活性

情報 Roscovitine (Seliciclib,CYC202)は有効な細胞週期卵白の依存性キナーゼ選択阻害剤、cdc2/ cyclin B、cdk2/ cyclin A、cdk2/ cyclin Eとcdk5/ p53に作用する時、IC50それぞれ0.65、0.7、0.7と0.16μM。
目標 Cdc2/cyclin B CDK2/cyclin A CDK2/cyclin E CDK5/p35
IC50 0.65 μM 0.7 μM 0.7 μM 0.16 μM [1]
In vitro試験 Roscovitine displays high efficiency and high selectivity towards some cyclin-dependent kinases with IC50 of 0.65, 0.7, 0.7 and 0.16 μM for cdc2/cyclin B, cdk2/cyclin A, cdk2/cyclin E and cdk5/p53, respectively. [1] Roscovitine reversibly inhibits the prophaselmetaphase transition in the micromolar range of starfish oocytes and sea urchin embryos, inhibits in vitro M-phase-promoting factor activity and in vitro DNA synthesis in Xenopus egg extracts, and suppresses the proliferation of mammalian cell lines with an average IC50 of 16 μM. [1] In mesangial cells, Roscovitine results in a dose-dependent reduction of CDK2 activity that at concentrations of 7.5, 12.5 and 25 mM, Roscovitine causes a 25, 50% and 100% decrease in CDK2 activity, respectively. [2] A recent study shows that Roscovitine inhibits cdk5 kinase activity, cell proliferation, multicellular development, and cdk5 nuclear translocation in Dictyostelium discoideum, without affecting the expression of cdk5 protein during axenic growth. [3]
In vivo試験 Roscovitine, at a dose of 50 mg/kg, significantly inhibits growth of The Ewing's sarcoma family of tumors (ESFT) xenografts. [4] Roscovitine enhances the antitumor effect of doxorubicin without increased toxicity with a mechanism that involves cell cycle arrest rather than apoptosis in nude mice bearing established MCF7 xenografts. [5]
臨床試験 Roscovitine is currently in Phase I clinical trial in patients with advanced solid tumors.
特集

推薦された実験操作 (公開の文献だけ)

キナーゼアッセイ: [1]

Enzymes Kinases activities are assayed at 30 °C in buffer C. Blank values are subtracted from the data and activities calculated as molar amount of phosphate incorporated in protein acceptor during a 10-minute incubation. Controls are performed with appropriate dilutions of DMSO. In a few cases, phosphorylation of the substrate is assessed by autoradiography after SDS/PAGE. p34cdc2/cyclin B is purified from M-phase starfish (M. glacialis) oocytes by affinity chromatography. It is assayed with 1 mg histone Hl/mL, in the presence of 15 μM [γ-32P]ATP (3000 Ci/mmol; 1 mCi/mL) in a final volume of 30 μL. After a 10-minute incubation at 30 °C, 25-μL aliquots of supernatant are spotted onto pieces of Whatman P81 phosphocellulose paper, and, after 20 seconds, the filters are washed five times (for at least 5 minutes each time) in a solution of 10mL phosphoric acid/L water. The wet filters are transferred into 6-mL plastic scintillation vials, 5 mL ACS scintillation fluid is added and the radioactivity measured in a Packard counter. The kinase activity is expressed as molar amount of phosphate incorporated in histone H1 during a 10-minutes incubation or as a percentage of maximal activity. p33cdk2/cyclin A and p33cdk2/cyclinE are reconstituted from extracts of sf9 insect cells infected with various baculoviruses. Cyclins A and E are fusion proteins with glutathione S-transferase and the complexes are purified on glutathione-agarose beads. Kinase activities are assayed with 1 mg/mL histone H1, in the presence of 15 μM [γ-32P]ATP, during 10 minutes, in a final volume of 30 μL, as described for the p34cdc2/cyclin B kinase. p33cdk5/p35 is purified from bovine brain, excluding the Mono S-chromatographic step. The active fractions from the Superose 12 column are pooled and concentrated to a final concentration of approximately 25 μg enzyme/mL. The kinase is assayed with 1 mg/mL histone HI in the presence of 15 μM [γ-32P]ATP, during 10 minutes in a final volume of 30 μL, as described for the p34cdc2/cyclin B kinase. p33cdk5/cyclin D1 is obtained from insect cell lysates. Cdk4 is a fusion protein with glutathione-S-transferase and the active complex is purified on glutathione-agarose beads. Its kinase activity is assayed with purified retinoblastoma protein (complexed with glutathione-S-transferase) in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. After a 15-minute incubation, 30 μL Laemmli sample buffer is added. The phosphorylated substrate is resolved by 10 % SDS/PAGE and analysed by autoradiography by overnight exposure to Hyperfilm MP and densitometry. p33cdk4/cyclinD 2 is obtained from insect cell lysates. It is assayed with purified retinoblastoma protein (complexed with glutathione-S-transferase) in the presence of 15 μM [γ-32P]ATP in a final volume of 30 μL. After a 30-minute incubation, 30 μL Laemmli sample buffer is added. The phosphorylated substrate is resolved by 10% SDS/PAGE and analysed by autoradiography by overnight exposure to Hyperfilm MP and densitometry. MAP kinase erkl (tagged with glutathione-S-transferase), is expressed in bacteria, purified on glutathione-agarose beads and assayed with 1 mg myelin basic protein/ml in the presence of 15 μM [γ-32P]ATP as described above for the p34cdc2cyclin B kinase. His-tagged erkl and erk2 are activated in vitro by mitogen-activated protein kinase kinase, purified (Ni-affinity and Mono Q) and assayed as described above during 10 minutes in a final volume of 30 μL. Protein kinase C isoforms are purified from baculovirus infected sf9 insect cells and assayed with 1 mg/mL protamine sulfate in the presence of 15 μM [γ-32P]ATP, during 10 minutes at 30 °C, in a final volume of 30 μL. Phosphorylated protamine sulfate is recovered on Whatman P81 phosphocellulose paper as described for the cdc2 kinase. The catalytic subunit of cAMP-dependent protein kinase, purified from bovine heart, is assayed with 1 mg histone Hl/ml, in the presence of 15 μM [γ-32P]ATP as described for the p34cdc2/cyclin B kinase. cGMP-dependent protein kinase, purified to homogeneity from bovine tracheal smooth muscle, is assayed with 1 mg histone Hl/mL, in the presence of 15 μM [γ-32P]ATP as described for the p34cdc2/cyclin B kinase. Casein kinase 2 is isolated from rat liver cytosol and assayed with 1 mg casein/mL and 15 μM [γ-32P]ATP. The substrate is spotted on Whatmann 3MM filters and washed with 10% (mass/vol.) trichloroacetic acid. Myosin light chain kinase, purified from chicken gizzard is assayed in the presence of 100 nM calmodulin, 100 μM CaCl2, 50 mM Hepes, 5 mM MgCI,, 1 mM dithiothreitol and 0.1 mg BSA/ml at pH 7.5 using a synthetic peptide based on the smooth-muscle myosin light-chain phosphorylation site and in the presence of 15 μM [γ-32P]ATP, in a final volume of 50 μL. Incorporation of radioactive phosphate is monitored on phosphocellulose filters as described above. ASK-γ, a plant homologue of GSK-3, is expressed as a glutathione-S-transferase fusion protein in Escherichia coli and purified on glutathione-agarose. ASK-γ kinase is assayed, for 10 minutes at 30 °C, with 5 μg myelin basic protein, in the presence of 15 μM [γ-32P]ATP in a final volume of 30 μL. The phosphorylated myelin basic protein is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase. Insulin receptor tyrosine kinase domain (CIRK-41) is overexpressed in a baculovirus system and purified to homogeneity. Its kinase activity is assayed, for 10 minutes at 30 °C, with 5 μg Raytide, in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. The phosphorylated Raytide is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase. c-src kinase is purified from infected Sf9 cells. The v-abl kinase is expressed in E. coli and affinity purified on IgG Affigel 10. Both kinases are assayed for 10 minutes at 30 °C, with 5 μg Raytide, in the presence of 15 μM [γ-32P]ATP, in a final volume of 30 μL. The phosphorylated Raytide is recovered on Whatman P81 phosphocellulose paper as described for the p34cdc2/cyclin B kinase.

細胞アッセイ: [1]

細胞系 Leukemia, non-small cell lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer
濃度 0.01 - 100 μM
処理時間 48 hours
方法 60 human tumour cell lines comprising nine tumor types are cultured for 24 hours prior to a 48-hour continuous exposure to 0.01-100 μM roscovitine. A sulforhodaminine B protein assay is used to estimate the cytotoxicity.

動物実験: [4]

動物モデル A4573 cells are injected s.c. into the right posterior flank of CD1 nu/nu mice.
製剤 Roscovitine is dissolved in either absolute methanol or DMSO and then diluted in 10% Tween 80, 20% N-N-dimethylacetamide, and 70% polyethylene glycol 400.
投薬量 ≤50 mg/kg
管理 Administered via i.p.
1

参考

化学情報

Download Roscovitine (Seliciclib,CYC202) SDF
分子量 354.45
化学式

C19H26N6O

CAS No. 186692-46-6
別名 N/A
溶解度 (25°C)
  • DMSO 71 mg/mL
  • 水 <1 mg/mL
  • エタノール 6 mg/mL
保管 2年 -20°C
6月-80°Cin DMSO
化学名 (R)-2-(6-(benzylamino)-9-isopropyl-9H-purin-2-ylamino)butan-1-ol

研究分野

カスタマーレビュー (4)


Click to enlarge
Rating
Source PNAS, 2011, 108, 8417. Roscovitine (Seliciclib,CYC202) purchased from Selleck
Method Western blotting, Fluorescent Microscopy, Confocal Imaging
Cell Lines Tumor xenografts
Concentrations 150 mg/kg
Incubation Time 15 d
Results R-roscovitine caused approximately 50% weight reduction of dissected tumor xenografts(Fig. A). Consistent with the in vitro observations, Western blot and immunohistochemistry analysis of tumor specimens showed suppressed ACTH and PCNA protein expression by R-roscovitine (Fig. B and C). R-roscovitine-treated mice exhibited more than 50% reduction in plasma ACTH levels ), and approximately 50% reduction in serum corticosterone levels.

Click to enlarge
Rating
Source Bai Bo University of Hong Kong. Roscovitine (Seliciclib,CYC202) purchased from Selleck
Method Oil-Red-O/SA-β-gal staining
Cell Lines ApoE-/- mice
Concentrations
Incubation Time 18 weeks
Results Chronic treatment with roscovitine attenuates the development of atherosclerosis in ApoE-/- mice.

Click to enlarge
Rating
Source The Journal of Neuroscience, 2012, 32(32):11050-11066. Roscovitine (Seliciclib,CYC202) purchased from Selleck
Method Western Blot/immunofluoresence
Cell Lines Isolated granular neurons
Concentrations 20 uM
Incubation Time 24 h
Results Application of roscovitine significantly suppressed T288 phosphorylation of Aurora-A associated with reduction of S251 phosphorylation of NDEL1, which was confirmed by Western blotting and immunocytochemistry.

Click to enlarge
Rating
Source PNAS, 2011, 108, 8417. Roscovitine (Seliciclib,CYC202) purchased from Selleck
Method Western blotting, Wst-1 proliferation assay, radioimmunoassays
Cell Lines AtT20 cells
Concentrations 0-20 µM
Incubation Time 24/48 h
Results Treatment with R-roscovitine led to decreased cell number by 24 h (Fig. A). Western blot analysis of protein extracts derived from R-roscovitine-treated cells revealed evidence for cell cycle arrest, including decreased cyclin E, increased p27Kip1, p57Kip2, and p21Cip1 expression, as well as reduced Thr821 phosphorylation of Rb (Fig. B). R-roscovitine treatment also induced senescent features by 48 h as evidenced by increased β -gal expression (Fig. C). Consistent with decreased cell viability, we detected decreased ACTH concentrations in culture medium derived from R-roscovitine-treated AtT20 cells (Fig. D). Western blot analysis of protein extracts derived from R-roscovitine-treated AtT20 cells showed suppressed ACTH expression (Fig. E). These results indicate that R-roscovitine targets cdk2/cyclin E-mediated cell cycle progression, and also inhibits corticotroph ACTH protein expression.

製品表彰状 (8)

  • MAPK phosphorylation of connexin 43 promotes binding of cyclin E and smooth muscle cell proliferation [Johnstone SR,et al. Circ Res 2012;111(2):201-11]

    PubMed: 22652908
  • Targeting zebrafish and murine pituitary corticotroph tumors with a cyclin-dependent kinase (CDK) inhibitor. [Liu NA, et al. Proc Natl Acad Sci U S A 2011;108(20):8414-9]

    PubMed: 21536883
  • Activation of Aurora-A Is Essential for Neuronal Migration via Modulation of Microtubule Organization. [Takitoh T, et al. J Neurosci 2012;32(32):11050-66]

    PubMed: 22875938
  • An RNAi-based screen reveals PLK1, CDK1 and NDC80 as potential therapeutic targets in malignant pleural mesothelioma. [Linton A, et al. Br J Cancer 2013;110(2):510-9]

    PubMed: 24327015
  • The cyclin-dependent kinase inhibitor p57(Kip2) is epigenetically regulated in carboplatin resistance and results in collateral sensitivity to the CDK inhibitor seliciclib in ovarian cancer. [Coley HM, et al. Br J Cancer 2012;106(3):482-9]

    PubMed: 22233925
  • An RNAi-based screen reveals PLK1, CDK1 and NDC80 as potential therapeutic targets in malignant pleural mesothelioma. [Linton A, et al. Br J Cancer 2013;10.1038/bjc.2013.731]

    PubMed: 24327015
  • A chrysin derivative suppresses skin cancer growth by inhibiting cyclin-dependent kinases. [Liu H, et al. J Biol Chem 2013;288(36):25924-37]

    PubMed: 23888052
  • Effect of Roscovitine on Intracellular Calcium Dynamics: Differential Enantioselective Responses. [Tamma G, et al. Mol Pharm 2013;10(12):4620-8]

    PubMed: 24168213

技術サポート&よくある質問(FAQ)

顧客がするかもしれない質問に対する答えは、指示を取り扱っている阻害剤で見つかります。話題は、貯蔵液(阻害剤と特別な注意を細胞ベースの分析法と動物のために必要とする問題を保存することは実験します)を準備する方法を含みます。

電話番号: +1-832-582-8158 Ext:3月曜日〜金曜日 9:00 AM–5:00 PM (米国中部標準時)

他の問い合わせをするならば、メッセージを残してください。

* 必須

Related CDK 阻害剤

  • PD0332991 HCl

    Palbociclib (PD-0332991) HClはCdksを抑制、Cdk4/ cyclin D1とCdk6/ cyclin D2に作用する時、IC50がそれぞれ11と16nMになる。

  • SNS-032 (BMS-387032)

    SNS-032 (BMS-387032)は、CDK2、CDK7とCDK9の新しくて、強力で、選択的なcdk阻害剤で、 IC50 がそれぞれ 38 nM、 62 nM 、 4 nMです。

  • Flavopiridol (Alvocidib)

    Flavopiridol (Alvocidib)は、CDK1、CDK2、CDK4、CDK5、CDK6とCDK9を目標としている汎cdk阻害剤で、IC50 がそれぞれ 30 nM、 170 nM、 100 nM、 170 nM、 80 nM と 20 nMです。

  • JNJ-7706621

    JNJ-7706621は新型有効の広い譜CDKとAuroraキナーゼ阻害剤、IC50が3-253nMになる。

  • PHA-793887

    PHA-793887は、CDK2、CDK5とCDK7の新しくて強力な阻害剤で、IC50 がそれぞれ 8 nM、 5 nM と 10 nMです。

  • AT7519

    AT7519は、新しい小分子です。それは、マルチ・サイクリン依存的なキナーゼ阻害剤で、CDK1/cyclin B、 CDK2/Cyclin A、 CDK3/Cyclin E、 CDk4/Cyclin D1、 CDk5/p35、 CDK6/Cyclin D3に作用すると、IC50が それぞれ210 nM、47 nM、360 nM、100 nM、13 nM、170 nM になる。

  • BS-181 HCl

    BS-181 HClはCDK7高度選択阻害剤、IC50が21nMになる。

  • Palbociclib (PD0332991) Isethionate

    Palbociclib (PD0332991) Isethionateは、非常に特定のCDK4CDK6の阻害剤で、 IC50 がそれぞれ11 nM と 16 nMです。

  • BMS-265246

    BMS-265246は、CDK1/cycBとCDK2/cycEのための強力で選択的なCDK1/CDK2阻害剤で、IC50 がそれぞれ 6 nM と 9 nMです。

  • AZD5438

    AZD5438は、サイクリン依存的なキナーゼ(cdk)1、2と9の強力な阻害剤で、IC50 がそれぞれ 16 nM、 6 nM、20 nMです。

最近見られたアイテム

Tags: Roscovitine (Seliciclib,CYC202)を買う | Roscovitine (Seliciclib,CYC202)供給者 | Roscovitine (Seliciclib,CYC202)を購入する | Roscovitine (Seliciclib,CYC202)費用 | Roscovitine (Seliciclib,CYC202)生産者 | オーダーRoscovitine (Seliciclib,CYC202) | Roscovitine (Seliciclib,CYC202)代理店
お問い合わせ