| 名稱 | KW-2449 |
| 描述 | KW-2449 is a multiple-targeted inhibitor, mostly for Flt3 (IC50: 6.6 nM), modestly effective to Bcr-Abl, FGFR1, and Aurora A; little inhibitory on PDGFRβ, IGF-1R, EGFR. |
| 細(xì)胞實驗 | Cell viability is determined by the sodium 3′-[1-(phenylaminocarbonyl)-3, 4-tetrazolium]-bis (4-methoxy-6-nitro) benzene sulfonic acid hydrate assay after incubation with or without KW-2449 for 72 hours at 37 °C. The number of viable cells is determined using the Cell Proliferation Kit II. For cell-cycle analysis, MOLM-13 and RS4;11 cells are treated with KW-2449. After 24, 48, and 72 hours of incubation at 37 °C, DNA contents are analyzed. Cell cycle distribution of K562, TCC-Y, and TCC/Ysr is analyzed 24 hours after treatment with KW-2449 or imatinib. (Only for Reference) |
| 激酶實驗 | FLT3 phosphorylation: Leukemia cells are washed in phosphate-buffered saline (PBS), then lysed by resuspending the cells in lysis buffer (20 mM Tris pH 7.4, 100 mM NaCl, 1% Igepal, 1 mM EDTA, 2 mM NaVO4, plus Complete protease inhibitor KW-2449 for 30 minutes while rocking. The extract is clarified by centrifugation at 1.6 × 104?g and the supernatant is assayed for protein (Bio-Rad). A 50-μg aliquot is removed as a whole-cell lysate for analysis of STAT5, and the remainder is used for immunoprecipitation with anti-FLT3. Anti-FLT3 antibody is added to the extract for overnight incubation, then protein A sepharose is added for 2 additional hours. Separate sodium dodecyl sulfate–polyacrylamide electrophoresis (SDS-PAGE) gels for whole-cell lysate and immunoprecipates are run in parallel. After transfer to Immobilon membranes, immunoblotting is performed with antiphosphotyrosine antibody (4 g10) to detect phosphorylated FLT3 or, for the whole-cell lysate gels, with a rat monoclonal antibody against phosphorylated STAT5 (residue Y694) then stripped and reprobed with anti-FLT3 antibody to measure total FLT3. Proteins are visualized using chemiluminescence, exposed on Kodak BioMax XAR film, developed, and scanned using a Bio-Rad GS800 densitometer. The concentration of KW-2449 for which the phosphorylation of FLT3 or STAT5 is inhibited to 50% of its baseline (IC50) is determined using linear regression analysis of the dose response curves. For direct analysis of FLT3 and STAT5 in circulating blasts, peripheral blood is collected in heparinized tubes and promptly chilled on ice. Samples are centrifuged for 10 minutes at 900 g, at 4 °C. The plasma is removed and stored frozen at ?80 °C. The buffy coat is carefully transferred to ice-cold PBS, layered onto chilled Ficoll-Hypaque, and centrifuged for 5 minutes at 600 g, at 4 °C. All subsequent steps are carried out at 4 °C. Mononuclear cells are collected and washed rapidly once in red blood cell lysis buffer (0.155 M NH4Cl, 0.01 M KHCO3, 0.1 mM EDTA), then washed once in PBS. Cells are then lysed as described for FLT3 and STAT5 analysis. |
| 體外活性 | 在MOLM-13移植瘤模型中,KW-2449（p.o.,14天）抗癌效果明顯,該作用存在劑量依賴性. |
| 體內(nèi)活性 | 在攜帶FLT3突變和抗Imatinib突變的白血病患者體內(nèi),KW-2449具有明顯活性,且獲得臨床研究授權(quán)。KW-2449可劑量依賴性地使FLT3和 STAT5的磷酸化程度降低。此外, KW-2449對ABL-T315I有顯著抑制效果（IC50：4 nM）。另一方面, KW-2449（1 μM）不影響PDGFRβ, IGF-1R, EGFR, 及多種絲/蘇氨酸激酶。作用于表達(dá)FLT3/ITD的白血病細(xì)胞和FLT3/KDM激活的以及過表達(dá)野生型FLT3的白血病細(xì)胞時,KW-2449對其生長均有明顯抑制作用。作用于MOLM-13細(xì)胞時,KW-2449劑量依賴性地抑制FLT3及其下游分子磷酸-STAT5的磷酸化。KW-2449還提高G1期百分?jǐn)?shù),并降低S期百分?jǐn)?shù),使得凋亡細(xì)胞數(shù)增高。KW-2449 可以使組成型活化的WT-FLT3 激酶去磷酸化,而對白血病細(xì)胞增殖無影響。 KW-2449會迅速被吸收,并轉(zhuǎn)化為一個主要的代謝產(chǎn)物M1。 |
| 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 : 55 mg/mL (165.46 mM), Sonication is recommended.
H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : 62 mg/mL (186.5 mM)
|
| 關(guān)鍵字 | Cluster of differentiation antigen 135 | Fibroblast growth factor receptor | inhibit | Fms like tyrosine kinase 3 | KW-2449 | Aurora Kinase | Inhibitor | FGFR | Bcr-Abl | KW 2449 | Apoptosis | FLT3 | CD135 |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Metronidazole | Ferulic Acid | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 神經(jīng)退行性疾病化合物庫 | 膜蛋白靶向化合物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |