| 名稱 | COTI-2 |
| 描述 | COTI-2, an orally available thiosemicarbazone, is an activator of mutant forms of the p53 protein with potential antineoplastic activity. |
| 細(xì)胞實(shí)驗(yàn) | COTI-2 is dissolved in 100% dimethyl sulfoxide stock solution and diluted in medium plus FBS such that final DMSO concentrations are 0.5 to 1.0% depending on the experiment.SHP-77 cells are cultured with various concentrations of COTI-2 for 48 h. Cells are then washed twice with 1× cold PBS and stained with Annexin V and 7AAD according to the manufacturer's instructions. Briefly, 5 μL of Annexin V and 7AAD are added to 1×105 cells and incubated for 15 min at room temperature in the dark. Then 400 μL of the 1× binding buffer (100 mM HEPES, pH 7.4, 140 mM NaCl, 2.5 mM CaCl2) is added to the cells. Finally, cells are analyzed using a flow cytometer. |
| 激酶實(shí)驗(yàn) | The interaction of COTI-2 with 227 kinases is tested using the AMBIT BIOSCIENCES KINOMESCAN assay. In brief, streptavidin-coated magnetic beads are treated with biotinylated small molecule ligands for 30 min at 25°C to generate affinity resins for kinase assays. The liganded beads are blocked with excess biotin and washed with blocking buffer (1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific binding. Binding reactions are assembled by combining phage lysates, liganded affinity beads, and COTI-2 in 1× binding buffer (20% SeaBlock, 0.17× PBS, 0.05% Tween 20, 6 mM DTT). All reactions are carried out in polystyrene 96-well plates that have been pre-treated with blocking buffer in a final volume of 0.1 mL. |
| 動(dòng)物實(shí)驗(yàn) | SHP-77 and HT-29 cells are injected in 50% matrigel into flanks of NCr-nu mice (2×106 cells per injection site) (n=5 mice per group). In the case of SHP-77 xenografts, treatment with COTI-2 begins prior to the appearance of palpable tumors. One day after injection of SHP-77 cells, animals receive 3 mg/kg of COTI-2 (once every two days, up to 38 days). Tumor sizes are estimated at 5, 10, 17, 24, and 38 days, by standard caliper measurements. In the case of HT-29 xenografts, the capacity of COTI-2 to suppress the growth of established tumors is assessed. HT-29 xenografts are allowed to grow to 200 mm3 before starting IP treatment with COTI-2 (10 mg/kg, 5 days a week for 7 weeks) or saline IP. Tumor growth is measured every 4 days by caliper measurement. |
| 體外活性 | COTI-2（72小時(shí)）顯著抑制了所有測(cè)試細(xì)胞系的增殖速率。在COLO-205、HCT-15和SW620細(xì)胞系中，COTI-2顯著抑制腫瘤細(xì)胞增殖。COTI-2的相對(duì)低濃度對(duì)所有測(cè)試的人類膠質(zhì)母細(xì)胞瘤細(xì)胞系（U87-MG、SNB-19、SF-268和SF-295）均有效。在SHP-77細(xì)胞中，COTI-2處理的IC50濃度導(dǎo)致40至47%的總細(xì)胞發(fā)生早期凋亡。 |
| 體內(nèi)活性 | 在HT-29人類結(jié)直腸癌異種移植模型中，COTI-2（10 mg/kg）顯著抑制腫瘤生長(zhǎng)。不僅在特定治療后時(shí)間點(diǎn)減少腫瘤體積，COTI-2還延長(zhǎng)腫瘤達(dá)到特定體積的時(shí)間。在SHP-77 SCLC異種移植模型中，COTI-2（3 mg/kg）同樣顯著抑制腫瘤生長(zhǎng)。COTI-2能在特定治療后時(shí)間點(diǎn)減少U87-MG腫瘤體積，并延長(zhǎng)U87-MG異種移植體在裸鼠體內(nèi)生長(zhǎng)所需的時(shí)間。僅用載體處理的對(duì)照組小鼠的腫瘤，5天內(nèi)就達(dá)到平均體積828 mm3，而用COTI-2處理的動(dòng)物腫瘤需要雙倍時(shí)間（10天）達(dá)到類似的平均體積（857 mm3）。無(wú)論采用何種給藥途徑，COTI-2均強(qiáng)效抑制OVCAR-3異種移植體的生長(zhǎng)。 |
| 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 : 5 mg/mL (13.64 mM)
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| 關(guān)鍵字 | inhibit | Inhibitor | HNSCC | low | anti-cancer | anti-proliferative | toxicity | TNBC | COTI-2 | MDM-2/p53 | Apoptosis | orally |
| 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 |
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