| 名稱 | COH29 |
| 描述 | COH29 (RNR Inhibitor COH29) is an orally available, aromatically substituted thiazole and inhibitor of the human ribonucleotide reductase (RNR), with potential antineoplastic activity. Upon oral administration, the RNR inhibitor COH29 binds to the ligand-binding pocket of the RNR M2 subunit (hRRM2) near the C-terminal tail. Inhibition of RNR activity decreases the pool of deoxyribonucleotide triphosphates available for DNA synthesis. The resulting decrease in DNA synthesis causes cell cycle arrest and growth inhibition. In addition, this agent may inhibit the nuclear enzyme poly (ADP-ribose) polymerase (PARP) 1, which prevents the repair of damaged DNA and causes both the accumulation of single and double strand DNA breaks and the induction of apoptosis. |
| 細胞實驗 | Cells is seeded into 96-well plates in 100 μL of complete medium at 2000 to 5000 cells per well, depending on the cell line's growth rate. After overnight incubation, test compound is added to each well at various concentrations in 50 μL of culture medium. After a further incubation for 96 hours at 37°C, fluorescein diacetate (final concentration: 10 mg/mL) and eosin Y [final concentration: 0.1% (w/v)] is added to each well, and the cells is incubated for an additional 20 minutes at 37°C. Cytotoxicity is assessed by Digital Imaging Microscopy System detection Viability is assessed using MTS [(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] as previously described[1]. |
| 激酶實驗 | 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 | COH29成功克服了Y細胞對吉西他濱和羥基脲的抗藥性。在NCI 60人類Y系列的大多數(shù)細胞系中，它強效抑制增殖，但對正常成纖維細胞或內(nèi)皮細胞幾乎無影響。NMR、定點突變技術(shù)和表面等離子體共振生物傳感器研究確認(rèn)COH29與提議的配體結(jié)合口袋結(jié)合，并為RRM1-RRM2四聚體結(jié)構(gòu)的組裝阻斷提供了證據(jù)[1]。 |
| 體內(nèi)活性 | COH29 (50/100 mg/kg, b.i.d., p.o.)在MOLT-4腫瘤移植模型中顯示出劑量依賴性的抑制增長效果，尤其在治療第12天時抑制效果顯著。在攜帶TOV11D腫瘤移植物的小鼠中，連續(xù)7天使用COH29 (200/300/400 mg/kg/天)同樣展示出劑量依賴性的腫瘤生長抑制效果。與對照組相比，腫瘤增長被顯著抑制[1]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 50 mg/mL (118.92 mM)
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| 關(guān)鍵字 | subunit | RNR Inhibitor COH-29 | RNR Inhibitor COH 29 | RNR | ribonucleotide | reductase | Inhibitor | inhibit | DNA/RNA Synthesis | COH-29 | COH29 | COH 29 | anticancer |
| 相關(guān)產(chǎn)品 | N-Nitrosodiethylamine | Acyclovir | Guanidine hydrochloride | Trimethoprim | Procaine | Pemetrexed | Folic acid | Resveratrol | 5-Fluorouracil | Temozolomide | Thymidine | Ribavirin |
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