| 名稱 | Dovitinib Dilactic Acid |
| 描述 | Dovitinib Dilactic Acid (Dovitinib (TKI-258) Dilactic Acid) is the Dilactic acid of Dovitinib, which is a multitargeted RTK inhibitor, mostly for class III (FLT3/c-Kit) with IC50 of 1 nM/2 nM, also potent to class IV (FGFR1/3) and class V (VEGFR1-4) RTKs with IC50 of 8-13 nM, less potent to InsR, EGFR, c-Met, EphA2, Tie2, IGFR1 and HER2. Phase 4. |
| 細胞實驗 | Cell viability is assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium (MTT) dye absorbance. Cells are seeded in 96-well plates at a density of 5 × 103 (B9 cells) or 2 × 104 (MM cell lines) cells per well. Cells are incubated with 30 ng/mL aFGF and 100 μg/mL heparin or 1% IL-6 where indicated and increasing concentrations of Dovitinib. For each concentration of Dovitinib, 10 μL aliquots of drug or DMSO diluted in culture medium is added. For drug combination studies, cells are incubated with 0.5 μM dexamethasone, 100 nM Dovitinib, or both simultaneously where indicated. To evaluate the effect of Dovitinib on growth of MM cells adherent to BMSCs, 104 KMS11 cells are cultured on BMSC-coated 96-well plates in the presence or absence of Dovitinib. Plates are incubated for 48 to 96 hours. For assessment of macrophage colony-stimulating factor (M-CSF)-mediated growth, 5 × 103 M-NFS-60 cells/well are incubated with serial dilutions of Dovitinib with 10 ng/mL M-CSF and without granulocyte-macrophage colony-stimulating factor (GM-CSF). After 72 hours cell viability is determined using Cell Titer-Glo Assay. Each experimental condition is performed in triplicate. (Only for Reference) |
| 激酶實驗 | In vitro kinase assays: The inhibitory concentration of 50% (IC50) values for the inhibition of RTKs by Dovitinib are determined in a time-resolved fluorescence (TRF) or radioactive format, measuring the inhibition by Dovitinib of phosphate transfer to a substrate by the respective enzyme. The kinase domains of FGFR3, FGFR1, PDGFRβ, and VEGFR1-3 are assayed in 50 mM HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid), pH 7.0, 2 mM MgCl2, 10 mM MnCl2 1 mM NaF, 1 mM dithiothreitol (DTT), 1 mg/mL bovine serum albumin (BSA), 0.25 μM biotinylated peptide substrate (GGGGQDGKDYIVLPI), and 1 to 30 μM adenosine triphosphate (ATP) depending on the Km for the respective enzyme. ATP concentrations are at or just below Km. For c-KIT and FLT3 reactions the pH is raised to 7.5 with 0.2 to 8 μM ATP in the presence of 0.25 to 1 μM biotinylated peptide substrate (GGLFDDPSYVNVQNL). Reactions are incubated at room temperature for 1 to 4 hours and the phosphorylated peptide captured on streptavidin-coated microtiter plates containing stop reaction buffer (25 mM EDTA [ethylenediaminetetraacetic acid], 50 mM HEPES, pH 7.5). Phosphorylated peptide is measured with the DELFIA TRF system using a Europium-labeled antiphosphotyrosine antibody PT66. The concentration of Dovitinib for IC50 is calculated using nonlinear regression with XL-Fit data analysis software version 4.1 (IDBS). Inhibition of colony-stimulating factor-1 receptor (CSF-1R), PDGFRα, insulin receptor (InsR), and insulin-like growth factor receptor 1 (IGFR1) kinase activity is determined at ATP concentrations close the Km for ATP. |
| 體外活性 | Dovitinib 對 FGF 刺激下生長的 WT 和 F384L-FGFR3 表達的 B9 細胞表現(xiàn)出強力抑制作用，具有 25 nM 的 IC50 值。此外，Dovitinib 還能抑制表達各種活化突變型 FGFR3 的 B9 細胞的增殖。有趣的是，對 Dovitinib 的敏感性在不同 FGFR3 突變之間的差異很小，IC50 值介于 70 到 90 nM 之間。僅含有載體的 IL-6 依賴性 B9 細胞（B9-MINV）對 Dovitinib 的抑制活性在高達 1 μM 的濃度下表現(xiàn)出抵抗性。Dovitinib 抑制 KMS11（FGFR3-Y373C）、OPM2（FGFR3-K650E）和 KMS18（FGFR3-G384D）細胞的細胞增殖，IC50 值分別為 90 nM（KMS11 和 OPM2）和 550 nM。Dovitinib 抑制 FGF 介導的 ERK1/2 磷酸化并在表達 FGFR3 的原發(fā)性 MM 細胞中引起細胞毒性。與未共培養(yǎng) BMSCs 的細胞相比，BMSCs 能為用 500 nM Dovitinib 處理的細胞提供輕微的抗性，前者的生長抑制率為 44.6%，而后者為 71.6%。Dovitinib 抑制由 M-CSF 驅動生長的小鼠骨髓增生性細胞系 M-NFS-60 的增殖，其 EC50 為 220 nM。[1] SK-HEP1 細胞對 Dovitinib 的處理導致細胞數(shù)量劑量依賴性下降，G2/M 期阻滯，G0/G1 和 S 期減少，抑制無錨生長，并阻斷 bFGF 引起的細胞運動。在 SK-HEP1 細胞中，Dovitinib 的 IC50 值約為 1.7 μM。Dovitinib 同時顯著減少了 SK-HEP1 和 21-0208 細胞中 FGFR-1、FGFR底物 2α（FRS2-α）和 ERK1/2 的基礎磷酸化水平，但對 Akt 無影響。在 21-0208 HCC 細胞中，Dovitinib 顯著抑制了 bFGF 引起的 FGFR-1、FRS2-α、ERK1/2 的磷酸化，但對 Akt 無影響。[2] |
| 體內活性 | Dovitinib在體內誘導細胞靜止和細胞毒性反應，導致表達FGFR3的腫瘤減小。[1] Dovitinib通過劑量和暴露程度依賴的方式，抑制腫瘤異種移植物中表達的目標受體酪氨酸激酶(RTKs)。Dovitinib強效地抑制六種HCC細胞系的腫瘤生長。抑制血管生成與FGFR/PDGFRβ/VEGFR2信號通路的失活相關。在原位模型中，Dovitinib強效地抑制原發(fā)性腫瘤生長和肺轉移，并顯著延長小鼠的存活時間。[2] Dovitinib的使用顯著抑制腫瘤生長并導致腫瘤回退，包括大型、已建立的腫瘤(500-1,000 mm3)。[3] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 83 mg/mL (144.96 mM), Sonication is recommended.
H2O : 64 mg/mL (111.77 mM), Sonication is recommended.
Ethanol : <1 mg/mL
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| 關鍵字 | VEGFR3/FLT4 | FLT3 | FGFR3 | FGFR1 | Dovitinib Dilactic Acid | c-Kit | cKit |
| 相關產品 | Amlexanox | Gilteritinib | Ribociclib | Lenvatinib | Chloramphenicol | Formononetin | Ferulic Acid | Pazopanib | Dasatinib | Sorafenib | Pexidartinib | Nintedanib esylate |