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| Dovitinib (TKI-258, CHIR-258)Multitargeted RTK inhibitor |
Sample solution is provided at 25 µL, 10mM.
Quality Control & MSDS
- View current batch:
- Purity = 98.62%
- COA (Certificate Of Analysis)
- HPLC
- NMR (Nuclear Magnetic Resonance)
- MSDS (Material Safety Data Sheet)
- Datasheet
Chemical structure
Related Biological Data
Related Biological Data
| Description | Dovitinib is a multitargeted inhibitor of RTK with IC50 values of 1 nM, 2 nM and 8 nM for FLT3, c-Kit and FGFR1, respectively. | |||||
| Targets | FLT3 | c-Kit | FGFR1 | VEGFR3/FLT4 | FGFR3 | VEGFR1/FLT1 |
| IC50 | 1 nM | 2 nM | 8 nM | 8 nM | 9 nM | 10 nM |
| Kinase experiment [1]: | |
Inhibitory activities | IC50 values for the inhibition of RTKs by CHIR-258 were determined in a time-resolved fluorescence (TRF) or radioactive format, measuring the inhibition by CHIR-258 of phosphate transfer to a substrate by the respective enzyme. The kinase domains of FGFR3, FGFR1, PDGFR-β and VEGFR1-3 were 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 were at or just below Km. For c-KIT and FLT3 reactions the pH was 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 were 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, 50mM HEPES, pH 7.5). |
| Cell experiment [1]: | |
Cell lines | Human multiple myeloma (MM) cell lines and B9 cells |
Preparation method | The solubility of this compound in DMSO is >36.4mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
Reaction Conditions | 100 nM CHIR-258; 48-96 h. |
Applications | Dovitinib is a receptor tyrosine kinases inhibitor. Dovitinib selectively inhibits the growth of human myeloma cell lines and B9 cells expressing wild-type (WT) or activated mutant FGFR3. Dovitinib also causes cytostatic and cytotoxic effects and inhibits downstream extracellular signal-regulated kinase (ERK) 1/2 phosphorylation. |
| Animal experiment [1]: | |
Animal models | 6- to 8-week-old female BNX mice bearing 3 ×107 KMS11 cells. |
Dosage form | 10, 30, or 60 mg/kg for 21 days by gavage. |
Preparation method | Dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM. For animal experiments: formulated in 5 mM citrate buffer. |
Applications | Dovitinib causes antitumor effect and inhibits tumor growths by 48%, 78.5%, and 94% in the 10 mg/kg, 30 mg/kg, and 60 mg/kg treatment arms, respectively. Weight loss, as a marker of significant toxicity, is not observed in any of the treatment groups. Dovitinib completely inhibits FGFR3 at the 60 mg/kg dose. CHIR-258 induces both cytostatic and cytotoxic responses. |
Other notes | Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal. |
References: [1]. Trudel S, Li ZH, Wei E, et al. CHIR-258, a novel, multitargeted tyrosine kinase inhibitor for the potential treatment of t(4;14) multiple myeloma. Blood, 2005, 105(7): 2941-2948. | |
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| Cas No. | 405169-16-6 | SDF | Download SDF |
| Synonyms | N/A | ||
| Chemical Name | (3Z)-4-amino-5-fluoro-3-[5-(4-methylpiperazin-1-yl)-1,3-dihydrobenzimidazol-2-ylidene]quinolin-2-one | ||
| Canonical SMILES | CN1CCN(CC1)C2=CC3=C(C=C2)NC(=C4C(=C5C(=NC4=O)C=CC=C5F)N)N3 | ||
| Formula | C21H21FN6O | M.Wt | 392.43 |
| Solubility | ≥36.35mg/mL in DMSO | Storage | Store at -20°C |
| Shipping Condition | Evaluation sample solution : ship with blue ice.All other available size:ship with RT , or blue ice upon request | ||
| General tips | For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months. | ||
Dovitinib (TKI258, CHIR-258) is a multitargeted receptor tyrosine kinase inhibitor against FLT3, KIT, FGFR, VEGFR, PDGFRα and PDGFRβ with IC50 of 1 nM, 2 nM, 8-9 nM, 8-13 nM , 210 nM and 27nM, respectively [1].
The viability of ZNF198-FGFR1 or BCR-FGFR1 cells was specifically inhibited by TKI258 with IC50 values of 150 nM or 90 nM. The phosphorylation of ERK and STAT5 genes was inhibited by TKI258 in dose dependent manner. TKI258 also specifically inhibited proliferation and survival of the FGFR1OP2-FGFR1-positive KG1 and KG1A cell lines, increasing levels of apoptosis [2]. In hepatocellular carcinoma (HCC) cells, the combination of TKI258 and tigatuzumab restored the sensitivity of HCC cells to TRAIL- and tigatuzumab-induced apoptosis. TKI258 inhibited phosphorylation of STAT3 and subsequently reduced the protein levels of Mcl-1, Survivin and Cylcin D1. Co-treatment of TRAIL and TKI258 increased the activity of SHP-1 [3]. Inhibition of FGFR3 with TKI258 decreased waldenstr m macroglobulinemia (WM) cell survival, increased apoptosis, and induced cell cycle arrest. TKI258 reduced the interaction of WM to bone marrow element, and reversed its proliferation. TKI258 had an additive effect with other drugs [4].
In vivo, the combination of tigatuzumab and TKI258 inhibited Huh-7 xenograft tumor growth [3]. TKI258 reduced WM tumor progression [4].
Reference:[1]. Trudel S, Li ZH, Wei E, Wiesmann M, Chang H, Chen C, Reece D, Heise C, Stewart AK. CHIR-258, a novel, multitargeted tyrosine kinase inhibitor for the potential treatment of t(4;14) multiple myeloma. Blood. 2005 Apr 1;105(7):2941-8.[2]. Chase A, Grand FH, Cross NC. Activity of TKI258 against primary cells and cell lines with FGFR1 fusion genes associated with the 8p11 myeloproliferative syndrome. Blood. 2007 Nov 15;110(10):3729-34.[3]. Chen KF, Chen HL, Liu CY, Tai WT, Ichikawa K, Chen PJ, Cheng AL. Dovitinib sensitizes hepatocellular carcinoma cells to TRAIL and tigatuzumab, a novel anti-DR5 antibody, through SHP-1-dependent inhibition of STAT3. Biochem Pharmacol. 2012 Mar 15;83(6):769-77.[4]. Azab AK, Azab F, Quang P, Maiso P, Sacco A, Ngo HT, Liu Y, Zhang Y, Morgan BL, Roccaro AM, Ghobrial IM. FGFR3 is overexpressed waldenstrom macroglobulinemia and its inhibition by Dovitinib induces apoptosis and overcomes stroma-induced proliferation. Clin Cancer Res. 2011 Jul 1;17(13):4389-99.
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如果是用转录出的cDNA做那跟平时就一样了。不需要特别的试剂盒、
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBRGreenMix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBRGreen，并且如果你那是ABI或者Stratagene的PCR如果用SYBRGreen还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
一、cDNA第二链的合成:
1. 第一链反应完成后,取2ul一链产物－20℃冰箱中保存，待电泳检测。其余的产物合并，混匀，然后顺序加入下列试剂(promega):
20ul 10×DNA Polymerase I buffer
6ul10mM dNTP(自己配制)
xuldd H2O
1ulRNase H(2U/ul)
10ul DNA Polymerase I(10U/ul)
总体系为200ul;
2. 混匀后,16℃反应2.5小时;
3. 70℃灭活10分钟;
4. 反应完成后,得到200ul cDNA第二链反应体系,将此体系置于冰上;
5．取2ul二链产物，同保存的一链产物一起电泳鉴定。同时上1kb ladder，确定双链的大小范围。
注：一链，二链的电泳图是smear，且二链稍比一链大一些。
二、双链cDNA末端补平:
1. 在第二链反应体系中,顺序加入下列试剂(promega):
6ul 10mM dNTP
2ul T4 DNA Polymerase(8.7U/ul)
2ul BSA(10mg/ml)
2. 稍微离心混匀反应物, 37℃反应至少30分钟,然后75℃灭活10分钟;
3. 加入等体积酚/氯仿/异戊醇,剧烈振荡后,常温下13000g离心5分钟;
4. 离心后,吸取上清于另一1.5ml eppendof管中,加入等体积氯仿,上下颠倒几次混匀后,常温下13000g离心5分钟;
5. 吸取上清至另一eppendof管,加入1/10V3M NaAc(PH5.2)和2.5V预冷的无水乙醇,混匀,-20℃放置过夜以沉淀双链cDNA;
6. 第二日,将昨日沉淀物在4℃,13000g离心60分钟以充分沉淀双链cDNA;
7.离心完毕,弃上清,加入1ml 70%乙醇洗涤沉淀,常温下13000g离心5分钟;
8.离心完毕,弃上清,干燥沉淀至无乙醇气味.
注：第3，第4步可以用PCR 纯化试剂盒代替。
PCR纯化试剂盒操作流程：
1．溶液PE使用前应加入适量体积95％－100％的乙醇，混匀。
2．向200ul二链补平产物中加入5倍体积的buffer PB，混匀。
3．加入spin column中，13000rpm离心1min。
4．加入0.75ml buffer PE，13000rpm离心1min。
5．13000rpm，再离心1min。
6．将spin column放入一新的离心管中，加入50ul buffer EB，静置10min。
7．13000rpm离心2min。
8．加入30ul buffer EB，静置10min。
9．13000rpm离心2min。
10．加入1/10体积3M的NaAc，2.5倍体积无水乙醇，混匀，－20℃沉淀过夜。
大家miRNA逆转录和qPCR的试剂盒用的是那个公司的啊，求推荐！
