Quizartinib (AC220)

 

Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	Quizartinib (AC220) is a second-generation FLT3 inhibitor for Flt3(ITD/WT) with IC50 of 1.1 nM/4.2 nM, 10-fold more selective for Flt3 than KIT, PDGFRα, PDGFRβ, RET, and CSF-1R. Phase 1/2.
Targets	Flt3ITD	Flt3wt
IC50	1.1 nM [1]	4.2 nM [1]
In vitro	AC220, a unique, potent and selective inhibitor of FLT3, has high affinity for FLT3 with a Kd value of 1.6 nM. AC220 inhibits the autophosphorylation of FLT3 in the human leukemia cell lines MV4-11 which harbor a homozygous FLT3-ITD mutation and is FLT3 dependent, and RS4;11 which expresses wild-type FLT3 with IC50 values of 1.1 nM and 4.2 nM, respectively. AC220 is the most potent cellular FLT3-ITD inhibitor, leading to the most significant inhibition of MV4-11 cell proliferation with IC50 of 0.56 nM compared to all other FLT3 inhibitors whose IC50 values range from 0.87 nM to 64 nM. AC220 has no inhibitory activity against the proliferation of A375 cells which harbor an activating mutation in BRAF and are not FLT3 dependent, indicating a large window between FLT3 inhibition and general cytotoxic effects. [1]
In vivo	Oral administration of AC220 (10 mg/kg) induces time-dependent inhibition of FLT3 autophosphorylation in the FLT3-ITD–dependent MV4-11 tumor xenograft mouse model; the inhibition being 90% at 2 hours and 40% at 24 hours. AC220 significantly extends survival in a mouse model of FLT3-ITD AML with doses as low as 1 mg/kg given orally once a day. Treatment with AC220 at 10 mg/kg for 28 days results in rapid and complete regression of tumors in all mice with no tumor regrowth during the 60-day posttreatment period. AC220 displays more significant efficacy compared to sunitinib treatment which causes tumors to shrink slowly and resume growth immediately upon discontinuation of treatment in all but one of the mice. [1]
Features	The most potent cellular FLT3-ITD inhibitor.

Protocol(Only for Reference)

Kinase Assay: ^[1]

Inhibition of FLT3 autophosphorylation

To measure inhibition of FLT3 autophosphorylation, MV4-11 or RS4;11 cells are cultured in low serum media (0.5% FBS) overnight and seeded at a density of 400 000 cells per well in a 96-well plate the following day. The cells are incubated with different concentrations of AC220 for 2 hours at 37 °C. To induce FLT3 autophosphorylation in RS4;11 cells, 100 ng/mL FLT3 ligand is added for 15 minutes after the 2-hour AC220 incubation. Cell lysates are prepared and incubated in 96-well plates precoated with a total FLT3 capture antibody. The coated plates are incubated with either a biotinylated antibody against FLT3 to detect total FLT3 or an antibody against phosphotyrosines to detect FLT3 autophosphorylation. In both cases, a SULFO-tagged streptavidin secondary antibody is used for electrochemiluminescence detection on the Meso Scale Discovery platform. The concentration of AC220 that inhibits FLT3-ITD or TLT3-WT autophosphorylation by 50% represents IC50 value

Cell Assay: ^[1]

Cell lines	MV4-11 and RS4;11 cells
Concentrations	Dissolved in DMSO, final concentration ~20 μM
Incubation Time	72 hours
Method	Cells are cultured overnight in low serum media (0.5% FBS), seeded in a 96-well plate at 40 000 cells per well and exposed to AC220 for 72 hours at 37 °C. Cell viability is measured using the Cell Titer-Blue Cell Viability Assa

Animal Study: ^[1]

Animal Models	Female NU/NU or severe combined immunodeficient mice implanted with MV4-11 cells
Dosages	~10 mg/kg
Administration	Oral gavage
Solubility	15% Captisol, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT00462761	Completed	Acute Myeloid Leukemia|Leukemia|Myelodysplastic Syndrome|AML|MDS	Ambit Biosciences Corporation	2007-01	Phase 1
NCT01390337	Recruiting	Leukemia, Myeloid, Acute	Astellas Pharma Inc|Ambit Biosciences Corporation	2011-10	Phase 1
NCT01468467	Recruiting	Leukemia, Myeloid, Acute,	Astellas Pharma Inc|Ambit Biosciences Corporation	2012-04	Phase 1
NCT01565668	Active, not recruiting	Leukemia, Myeloid, Acute	Astellas Pharma Inc|Astellas Pharma Global Development, Inc.|Ambit Biosciences Corporation	2012-04	Phase 2

Chemical Information

Download Quizartinib (AC220) SDF

Chemical Name	Urea, N-[5-(1,1-dimethylethyl)-3-isoxazolyl]-N-[4-[7-[2-(4-morpholinyl)ethoxy]imidazo[2,1-b]benzothiazol-2-yl]phenyl]-

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.56067	5.6067	11.2134	16.8201

Customer Reviews (1)

Click to enlarge	Rating
	Source	Quizartinib (AC220) purchased from Selleck
	Method	MTT Assay
	Cell Lines	epidermoid carcinoma cell line
	Concentrations	0.75-3 μM
	Incubation Time
	Results	Effect of AC220 on the sensitivity of KB-C2 cells to paclitaxel.

Product Citations (5)

• 
  

  MUC1-C oncoprotein promotes FLT3 receptor activation in acute myeloid leukemia cells. [Liu S, et al. Blood 2013;10.1182/blood-2013-04-493858]

  
  PubMed: 24282218


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  A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets. [Uitdehaag JC, et al. Br J Pharmacol 2012;166(3):858-76]

  
  PubMed: 22250956


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  The FLT3 Inhibitor Quizartinib Inhibits ABCG2 at Phar macologica lly Relevant Concen trations, with Implica tions for Both Chem osensit ization and Adverse Drug Interactions. [Bhullar J, et al. PLoS One 2013;8(8): e71266]

  
  PubMed: 23967177