| 名稱 | Rebastinib |
| 描述 | DCC-2036 (Rebastinib (DCC-2036)) is a conformational control Bcr-Abl inhibitor for Abl1(WT, IC50: 0.8 nM) and Abl1(T315I, IC50: 4 nM), also inhibits LYN, SRC, HCK, FGR, FLT3, KDR, and Tie-2, and low activity to c-Kit. Rebastinib aimed at the Angiopoietin2-Tie2 pathway. |
| 細(xì)胞實(shí)驗(yàn) | Ba/F3 cells or primary Ph+ leukemia cells are plated in triplicate in 96-well plates containing test compounds. After 72 hours, viable cells are quantified by Resazurin or MTT assay. Cells are diluted in medium to be added to each well of a 96-well tissue culture-treated plate. All cells are incubated overnight and maintained in a humidified atmosphere at 37 °C and 5% CO2. Cells are treated the following day. Serum-free medium is used during treatment with DCC-2036. MTT is used to assess the viability of cells following treatment. Aliquots of 20 mL of stock MTT solution are added to each well containing 200 mL of medium (10% final solution) and incubated with the cells for 2 hours. Following incubation the medium is removed and 200 mL of dimethylsulfoxide added to solubilize the formazan crystals. The absorbance is read on the plate reader at 550 and 690 nm. A subtraction analysis of the dual wavelength is performed (D550 to D690) to increase accuracy of the measuremen(Only for Reference) |
| 激酶實(shí)驗(yàn) | Assay of Abl1 kinase isoforms and determination of inhibitor potency: Activity of u-Abl1native is determined by following the production of ADP from the kinase reaction through coupling with the pyruvate kinase/lactate dehydrogenase system. In this assay, the oxidation of NADH (measured as a decreased A340 nM) is continuously monitored spectrophotometrically. The final reaction mixture (100 μL, in a 384-well Corning plate) is prepared as follows: An Abl1 kinase/coupled assay components mixture is prepared containing u-Abl1 kinase (1 nM), Abltide (EAIYAAPFAKKK, 0.2 mM), MgCl2 (9 mM), pyruvate kinase (~ 4 units), lactate dehydrogenase (~ 0.7 units), phosphoenol pyruvate (1 mM), and NADH (0.28 mM) in 90 mM Tris containing 0.1 % octyl-glucoside and 1 % DMSO, pH 7.5. Separately, an inhibitor mixture is prepared containing DCC-2036 serially diluted 3-fold in DMSO followed by dilution into buffer composed of 180 mM Tris, pH 7.5, containing MgCl2 (18 mM) and 0.2 % octyl-glucoside. Fifty μL of the inhibitor mixture is mixed with 50 μL of the above Abl1 kinase/coupled assay components mixture, which is then incubated at 30 °C for 2 hours before 2 μL of 25 mM ATP (500 μM, final) is added to start the reaction. The reaction is recorded every 2 minutes for 2.5 hours at 30 °C on a Polarstar Optima or Synergy2 plate reader. Reaction rate (slope) is calculated using the 1 to 2 hour time frame with reader's software. Percent inhibition is obtained by comparison of reaction rate with that of a DMSO control. IC50 values are calculated from a series of percent inhibition values determined at a range of inhibitor concentrations using GraphPad Prism. The kinase assay for Abl1T315I, p-Abl1native or Abl1H396P is assayed the same as above except that 2.2 nM Abl1T315I, 1 nM p-Abl1 native or 1.3 nM Abl1H396P is used. The above assay format is also used for kinases other than Abl1 with the exception of TIE2, for which a fluorescence polarization/Transcreener format is used.The assay conditions are the same as described above except that PolyE4Y (final 1 mg/mL) is used as the substrate and one hour preincubation is used. |
| 體外活性 | 給在攜帶Ba/F3-BCR-ABL1T315I白血病細(xì)胞的小鼠移植瘤模型中,DCC-2036（100 mg/kg/day,p.o.）對(duì)BCR-ABL1信號(hào)具有明顯抑制效果,并顯著延長(zhǎng)小鼠壽命. |
| 體內(nèi)活性 | DCC-2036對(duì)野生型或突變型BCR-ABL1的Ba/F3細(xì)胞有抗增殖活性（IC50：2-150 nM）。此外,DCC-2036對(duì)Ph+細(xì)胞系K562增殖也有抑制作用（IC50：5.5 nM）,并有效誘導(dǎo)表達(dá)BCR-ABL1的Ba/F3和K562細(xì)胞凋亡。 通過(guò)對(duì)CML細(xì)胞系的顯著抑制（與非CML白血病細(xì)胞系相比）,DCC-2036可選擇性抑制BCR-ABL陽(yáng)性細(xì)胞。DCC-2036對(duì)純化的未磷酸化和磷酸化ABL1（IC50：0.8/2 nM）、未磷酸化和磷酸化突變型ABL1T315I（IC50：1.4/5 nM）及激活環(huán)突變ABL1H396P（IC50：4 nM）呈較強(qiáng)的非ATP競(jìng)爭(zhēng)性抑制作用。DCC-2036還抑制SRC家族激酶SRC/LYN/FGR/HCK及受體 TKs KDR/FLT3/TIE2（IC50：34/29/38/40/4/2/6 nM）。 |
| 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 | Ethanol : 13 mg/mL (23.5 mM)
DMSO : 5.53 mg/mL (10 mM), Sonication is recommended.
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| 關(guān)鍵字 | CD135 | Apoptosis | inhibit | Bcr-Abl | Fms like tyrosine kinase 3 | Inhibitor | FLT3 | Src | Cluster of differentiation antigen 135 | Rebastinib |
| 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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