多韋替尼作用機(jī)制 - Medchemexpress - MCE中國

1、Product Data SheetDovitinibCat. No.: HY-50905CAS No.: 405169-16-6分式: CHFNO分量: 392.43作靶點(diǎn): c-Kit; FLT3; FGFR; VEGFR; PDGFR作通路: Protein Tyrosine Kinase/RTK儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 25 mg/mL (63.71 mM; Need ultrasonic and warming)SolventMass1

2、mg 5 mg 10 mgConcentration制備儲備液1 mM 2.5482 mL 12.7411 mL 25.4823 mL5 mM 0.5096 mL 2.5482 mL 5.0965 mL10 mM 0.2548 mL 1.2741 mL 2.5482 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲存時(shí)，請?jiān)?6 個(gè)內(nèi)使，-20C 儲存時(shí)，請?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解案都?/p>

3、先按照 In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實(shí)驗(yàn)結(jié)果的可靠性，澄 的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (6.37 mM); Clear solution此案可獲得 2.5 mg/mL (6.37 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 10

4、0 L 25.0 mg/mL 的澄 DMSO 儲備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (6.37 mM); Clear solution此案可獲得 2.5 mg/mL (6.37 mM，飽和度未知) 的澄清溶液。Page 1 of 2 www.MedChemE以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄均勻。DMSO 儲備液加

5、到 900 L 20% 的 SBE-CD 理鹽溶液中，混合3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (6.37 mM); Clear solution此案可獲得 2.5 mg/mL (6.37 mM，飽和度未知) 的澄 溶液，此案不適于實(shí)驗(yàn)周 期在半個(gè)以上的實(shí)驗(yàn)。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Dovitinib (TKI258; CHIR-258)是多靶點(diǎn)的酪氨酸激酶抑制劑，抑制FLT3，c

6、-Kit，F(xiàn)GFR1/3，VEGFR1/2/3 和 PDGFR/ 的 IC50 值分別為1，2，8/9，10/13/8，27/210 nM。IC & Target FGFR1 FGFR3 VEGFR1 VEGFR28 nM (IC50) 9 nM (IC50) 1 nM (IC50) 13 nM (IC50)VEGFR3 PDGFR PDGFR FLT38 nM (IC50) 27 nM (IC50) 210 nM (IC50) 1 nM (IC50)c-Kit2 nM (IC50)體外研究 Dovitinib potently inhibits the FGF-stimulated grow

7、th of WT and F384L-FGFR3-expressing B9 cells with IC50 values of25 nM. B9-MINV cells are resistant to the inhibitory activity of Dovitinib at concentrations up to 1 M. Dovitinibinhibits cell proliferation of KMS11 (FGFR3-Y373C), OPM2 (FGFR3-K650E), and KMS18 (FGFR3-G384D) cells with IC50of values of

8、 90 nM (KMS11 and OPM2) and 550 nM, respectively1. Dovitinib significantly reduces the basalphosphorylation levels of FGFR-1, FGFR substrate 2 (FRS2-) and ERK1/2 but not Akt in both SK-HEP1 and 21-0208cells2. Dovitinib enhances the BMP-2-induced alkaline phosphatase (ALP) induction, which is a repre

9、sentative markerof osteoblast differentiation. Dovitinib also stimulates the translocation of phosphorylated Smad1/5/8 into thenucleus and phosphorylation of mitogen-activated protein kinases, including ERK1/2 and p383. Dovitinib stronglyinhibits both the interaction of TNIK with ATP (Ki, 13 nM) and

10、 the activation of Wnt signaling effectors such as -catenin and TCF4. Dovitinib also induces caspase-dependent apoptosis in IM-9 cells without significant cytotoxicity inPBMCs4.體內(nèi)研究 Dovitinib (10 mg/kg, 30 mg/kg, 60 mg/kg, p.o.) shows significant antitumor effect in the KMS11-bearing mice model,and

11、the growth inhibition is 48%, 78.5%, and 94% in the 10 mg/kg, 30 mg/kg, and 60 mg/kg treatment arms,respectively, compared with the placebo-treated mice1. Dovitinib (50 and 75 mg/kg) results in 97% and 98% tumorgrowth inhibition, respectively, and the maximal efficacy is at 50 mg/kg2.PROTOCOLKinase

12、Assay 1 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 asubstrate by the respective enzyme. The kinase domains of FGFR

13、3, FGFR1, PDGFR-, and VEGFR1-3 are assayed in 50mM HEPES (N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid), pH 7.0, 2 mM MgCl2, 10 mM MnCl2 1 mM NaF, 1mM dithiothreitol (DTT), 1 mg/mL bovine serum albumin (BSA), 0.25 M biotinylated peptide substrate(GGGGQDGKDYIVLPI), and 1 to 30 M adenosine triph

14、osphate (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 M ATPPage 2 of 3 www.MedChemEin the presence of 0.25 to 1 M biotinylated peptide substrate (GGLFDDPSYVNVQNL). Reactions are incub

15、ated atroom temperature for 1 to 4 hours and the phosphorylated peptide captured on streptavidin-coated microtiter platescontaining 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-la

16、beled antiphosphotyrosineantibody PT66. The concentration of Dovitinib for IC50 is calculated using nonlinear regression with XL-Fit dataanalysis software version 4.1. Inhibition of colony-stimulating factor-1 receptor (CSF-1R), PDGFR-, insulin receptor(InsR), and insulin-like growth factor receptor

17、 1 (IGFR1) kinase activity is determined using the IC50 Profiler Expressservice.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cell viability is assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium (MTT) dye absorbance according to

18、 themanufacturers instructions. Cells are seeded in 96-well plates at a density of 5000 (B9 cells) or 20 000 (MM cell lines)cells per well. Cells are incubated with 30 ng/mL aFGF and 100 g/mL heparin or 1% IL-6 where indicated andincreasing concentrations of Dovitinib. For each concentration of Dovi

19、tinib, 10-L aliquots of drug or DMSO dilutedin culture medium is added. For drug combination studies, cells are incubated with 0.5 M dexamethasone, 100 nMDovitinib, or both simultaneously where indicated. To evaluate the effect of Dovitinib on growth of MM cellsadherent to BMSCs, 10 000 KMS11 cells

20、are cultured on BMSC-coated 96-well plates in the presence or absence ofDovitinib. Plates are incubated for 48 to 96 hours. For assessment of macrophage colony-stimulating factor (M-CSF)-mediated growth, 5000 M-NFS-60 cells/well are incubated with serial dilutions of CHIR-25 ZS8 with 10 ng/mL M-CSFa

21、nd without granulocyte-macrophage colony-stimulating factor (GM-CSF). After 72 hours cell viability is determinedusing Cell Titer-Glo Assay. Each experimental condition is performed in triplicate.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Th

22、e xenograft mouse model is prepared as previously described.27 Briefly, 6- to 8-week-old female BNX mice areAdministration 1 inoculated subcutaneously into the right flank with 3107 KMS11 cells in 150 L IMDM, together with 150 LMatrigel basement membrane matrix. Treatment is initiated when tumors re

23、ach volumes of 200 mm3 at which timemice are randomized to receive 10, 30, or 60 mg/kg Dovitinib or 5 mM citrate buffer. Dosing is performed daily for 21days by gavage. Eight to 10 mice are included in each treatment group. Caliper measurements are performed twiceweekly to estimate tumor volume, usi

24、ng the formula: 4/3 (width/2)2 (length/2). One-way analysis of variance isused to compare differences between vehicle- and Dovitinib-treated groups.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Sci Transl Med. 2018 Jul 18;10(450). pii: eaa

25、q1093. Theranostics. 2018 Jul 30;8(15):4262-4278. Front Cell Dev Biol. 2020 May 7;8:287. Biochemistry for Health, NOVA University of Lisbon. 2019 Jul. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Trudel S, et al. CHIR-258, a novel, multitargeted tyrosine kinase inhibitor for the potential trea