Flavopiridolは、サイクリン依存的なkianses（CDK）を妨げるために、ATPと争いまして、 CDK1, CDK2, CDK4、CDK6 に作用すると、IC50が40 nMになり、CDK7に作用すると、IC50が 300 nMになる。
| 情報 | Flavopiridolは、サイクリン依存的なkianses（CDK）を妨げるために、ATPと争いまして、 CDK1, CDK2, CDK4、CDK6 に作用すると、IC50が40 nMになり、CDK7に作用すると、IC50が 300 nMになる。 | |||||
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| 目標 | CDK1 | CDK2 | CDK4 | CDK6 | CDK7 | |
| IC50 | 40 nM | 40 nM | 40 nM | 40 nM | 300 nM [1] | |
| In vitro試験 | Flavopiridol is initially found to inhibit the epidermal growth factor receptor and protein kinase A (IC50 = 21 and 122 μM). Flavopiridol is later shown to inhibit cell proliferation, at more physiologically relevant concentrations (IC50 = 66 nM) when Flavopiridol is tested in the National Cancer Institute Development Therapeutics Program panel of 60 human tumor cell lines. [1] Flavopiridol induces G1 arrest with inhibition of CDK2 and CDK4 in human breast carcinoma cells in a time and concentration dependent manner. [2] Short time treatment of Flavopiridol (~12 hours) induce apoptosis in hematopoietic cell lines including SUDHL4, SUDHL6 (B-cell lines), Jurkat and MOLT4 (T-cell lines ), and HL60 (myeloid). [3] In the clonogenic assay, Flavopiridol functions as a highly potent cytotoxic compound with a mean IC70 with 8 ng/mL in 23 human tumor models. [4] A recent study shows Flavopiridol treatment induces a substantial AKT-Ser473 phosphorylation in human glioblastoma T98G cell line. [5] | |||||
| In vivo試験 | At the maximal tolerated dose of 10 mg/kg/day administered p.o. on days 1-4 and 7-11, Flavopiridol effects tumor regression in PRXF1337 and tumor stasis lasting for 4 weeks in PRXF1369. [4] After treatment with 7.5 mg/kg Flavopiridol bolus intravenous (IV) or intraperitoneal on each of 5 consecutive days, 11 out of 12 advanced stage subcutaneous (s.c.) human HL-60 xenografts undergo complete regressions, and animals remain disease-free several months after one course of Flavopiridol treatment. SUDHL-4 s.c. lymphomas treated with flavopiridol at 7.5 mg/kg bolus IV for 5 days undergo either major (two out of eight mice) or complete (four out of eight mice) regression, with two animals remaining disease-free for more than 60 days. The overall growth delay is 73.2%. [6] | |||||
| 臨床試験 | ||||||
| 特集 | ||||||
| Recombinant CDKs Kinase Reactions | CDKs activities are determined in microtiter plates as follows. Forty μg Gst-Rb are mixed with different amounts of Flavopiridol and unlabeled ATP. Reactions are then started by the addition of an ammonium sulfate cut of the S100 fraction obtained from insect cells expressing recombinant human CDKs. The final reaction conditions are 10 mM MgCl2, 50 mM Tris-HCl (pH 7.5), and 1 mM DTT. The final concentration of ATP is adjusted accordingly. Radiolabeled ATP is used as a phosphoryl donor. The reaction is carried out for 2.5 minutes at 30 °C after addition of enzyme and then terminated with the addition of EDTA. The Gst-Rb is then captured with glutathione-Sepharose and the incorporated radioactivity is determined by liquid scintillation counting. |
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| 細胞系 | SUDHL4, SUDHL6, Jurkat, MOLT4, and HL60 |
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| 濃度 | 0, 100 500, 5000 nM |
| 処理時間 | 14 hours |
| 方法 | Cells grown at a density of 1 × 106 cells/mL are exposed to Flavopiridol for different concentrations and time periods. DNA is extracted. Briefly, cells are washed once with cold phosphate-buffered saline (PBS) and lysed with 3 mL lysis buffer (5 mM Tris-HCL [pH 7.5]; 20 mM EDTA; 0.5% Triton X-100) for 15 minutes at 4 °C. The chromatin of the cell lysates is isolated by centrifugation (20 minutes at 26,000g, 4 °C). The supernatants containing small DNA fragments are extracted sequentially with phenol, phenol:chloroform (1:1), and chloroform. Nucleic acids are precipitated in 0.5 M NaCl, 90% ethanol at -20 °C overnight. RNA is then digested by bovine RNAaseA (60 μg/mL). After sequential reextraction and reprecipitation, DNA is dissolved in 10 mM Tris-HCL (pH 7.5), 1 mM EDTA, 0.5% sodium dodecyl sulfate (SDS) before electrophoresis on 1.6% agarose gel. |
| 動物モデル | human prostate cancer xenografts, PRXFI337 and PRXFI369, grown s.c. in nude mice |
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| 製剤 | Flavopiridol is dissolved in water. |
| 投薬量 | 10 mg/kg/d |
| 管理 | Flavopiridol is administered p.o. on days 1-4 and 7-11. |
| 分子量 | 438.3 |
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| 化学式 | C21H20ClNO5.HCl |
| CAS No. | 131740-09-5 |
| 別名 | HMR-1275, L86-8275 |
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溶解度 (25°C)
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| 保管 | 2年 -20°C粉 |
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| 2週4°Cin DMSO | |
| 6月-80°Cin DMSO | |
| 化学名 | 2-(2-chlorophenyl)-5,7-dihydroxy-8-((3S,4R)-3-hydroxy-1-methylpiperidin-4-yl)-4H-chromen-4-one hydrochloride |
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| Source | PNAS, 2011.May, 108:8417. Flavopiridol (Alvocidib) HCl purchased from Selleck | |
| Method | Fluorescent Microscopy/Confocal Imaging | |
| Cell Lines | Tg:Pomc-Pttg embryo, Pomc-eGFP embryo | |
| Concentrations | 50 μM | |
| Incubation Time | 18-48 h | |
| Results | Although flavopiridol retarded early embryonic development before corticotroph ontogeny occurred, in vivo treatment of zebrafish embryos with R-roscovitine, olomoucine, PD-0332991, and CAY10572 starting at 18 hpf caused no apparent growth defect by 40 hpf. Strikingly, R-roscovitine-treated embryos exhibited approximately 40% reduction in pituitary POMC-eGFP expression compared with controls. |
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), 8414-8419]
PubMed: 21536883Mechanisms of action of a dual Cdc7/Cdk9 kinase inhibitor against quiescent and proliferating CLL cells. [Natoni A, et al. Mol Cancer Ther 2011;10(9), 1624-1634]
PubMed: 21768328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。
* 必須
PD0332991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。
Roscovitine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。
Flavopiridol (Alvocidib, HMR-1275, L86-8275)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はCDK7高度選択阻害剤、IC50が21nMになる。
PD 0332991は、非常に特定のCDK4 と CDK6の阻害剤で、 IC50 がそれぞれ11 nM と 16 nMです。
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。
in vivo invitation
