Home JAK/STAT JAK inhibitor AT9283

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AT9283 JAK inhibitor

Cat.No.S1134

AT9283 is a potent JAK2/3 inhibitor with IC50 of 1.2 nM/1.1 nM in cell-free assays; also potent to Aurora A/B, Abl1(T315I).
AT9283 JAK inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 381.43

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Quality Control

Batch: Purity: 99.82%
99.82

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HCT116 Cytotoxicity assay 10 to 14 days Cytotoxicity against human HCT116 cells assessed as number of colonies after 10 to 14 days by colony forming assay, IC50=0.012μM 19143567
HCT116 Function assay 10 mg/kg Cmax in BALB/c mouse bearing human HCT116 cells at 10 mg/kg, po, Cmax=0.45μM 19143567
HCT116 Function assay 5 mg/kg Cmax in BALB/c mouse bearing human HCT116 cells at 5 mg/kg, iv, Cmax=4.9μM 19143567
HCT116 Function assay 20 mg/kg Cmax in BALB/c mouse bearing human HCT116 cells at 20 mg/kg, ip, Cmax=8.4μM 19143567
HT-29 Antitumor assay 72 hrs Antitumor activity against human HT-29 cells after 72 hrs by MTT assay, IC50=0.383μM 23664099
A549 Antitumor assay 72 hrs Antitumor activity against human A549 cells after 72 hrs by MTT assay, IC50=0.512μM 23664099
LoVo Antitumor assay 72 hrs Antitumor activity against human LoVo cells after 72 hrs by MTT assay, IC50=0.553μM 23664099
K562 Antitumor assay 72 hrs Antitumor activity against human K562 cells after 72 hrs by MTT assay, IC50=1.6μM 23664099
U937 Antitumor assay 72 hrs Antitumor activity against human U937 cells after 72 hrs by MTT assay, IC50=6.7μM 23664099
BL21 (DE3) Function assay 30 mins Inhibition of His6-tagged MELK catalytic domain (1 to 340 residues) (unknown origin) expressed in Escherichia coli BL21 (DE3) cells using Bcl-GL as substrate measured after 30 mins in presence of [gamma32P]ATP by liquid scintillation counting method, IC50=0.685μM 28351607
Sf9 Function assay Binding affinity to N-terminal TEV-cleavable hexa-histidine tagged human JAK2 JH1 domain (840 to 1132 residues) expressed in baculovirus-infected Sf9 cells by ITC assay, Kd=0.011μM 28626521
Sf9 Function assay Binding affinity to C-terminal thrombin-cleavable hexa-histidine tagged human JAK2 JH2 pseudokinase domain (536 to 812 residues) W659A/W777A/F794H mutant expressed in baculovirus-infected Sf9 cells by ITC assay, Kd=1.323μM 28626521
Sf9 Function assay 10 uM 60 mins Displacement of BODIPY-ATP from C-terminal thrombin-cleavable hexa-histidine tagged human JAK2 JH2 pseudokinase domain (536 to 812 residues) W659A/W777A/F794H mutant expressed in baculovirus-infected Sf9 cells at 10 uM after 60 mins by high-throughput flu 28626521
HCT116 Function assay Inhibition of Aurora B kinase in human HCT116 cells assessed as reduction in polyploid phenotype, IC50=0.03μM 28918096
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
U-2 OS qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells 29435139
Saos-2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells 29435139
fibroblast cells qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for control Hh wild type fibroblast cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-MC cells 29435139
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 76 mg/mL (199.25 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 25 mg/mL

Water : Insoluble

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Mass Concentration Volume Molecular Weight
Dilution Calculator Molecular Weight Calculator

In vivo
Batch:

In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
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Chemical Information, Storage & Stability

Molecular Weight 381.43 Formula

C19H23N7O2

 Storage (From the date of receipt)
CAS No. 896466-04-9 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles C1CC1NC(=O)NC2=C(NN=C2)C3=NC4=C(N3)C=C(C=C4)CN5CCOCC5

Mechanism of Action

Targets/IC50/Ki
JAK3
(Cell-free assay)
1.1 nM
JAK2
(Cell-free assay)
1.2 nM
Aurora A
(Cell-free assay)
~3.0 nM
Aurora B
(Cell-free assay)
~3.0 nM
Abl1 (T315I)
(Cell-free assay)
4 nM
GSK-3β
(Cell-free assay)
1 nM-10 nM
FGFR2
(Cell-free assay)
1-10 nM
VEGFR3/FLT4
(Cell-free assay)
1 nM-10 nM
Mer
(Cell-free assay)
1 nM-10 nM
RET
(Cell-free assay)
1 nM-10 nM
RSK2
(Cell-free assay)
1 nM-10 nM
RSK3
(Cell-free assay)
1 nM-10 nM
TYK2
(Cell-free assay)
1 nM-10 nM
YES
(Cell-free assay)
1 nM-10 nM
Abl (Q252H)
(Cell-free assay)
10 nM-30 nM
DRAK1
(Cell-free assay)
10 nM-30 nM
FGFR1
(Cell-free assay)
10 nM-30 nM
FGFR1 (V561M)
(Cell-free assay)
10 nM-30 nM
FGFR2 (N549H)
(Cell-free assay)
10 nM-30 nM
FGFR3
(Cell-free assay)
10 nM-30 nM
VEGFR1/FLT1
(Cell-free assay)
10 nM-30 nM
FLT3
(Cell-free assay)
10 nM-30 nM
PDGFRα (D842V)
(Cell-free assay)
10 nM-30 nM
PDK-1
(Cell-free assay)
10 nM-30 nM
PKCμ
(Cell-free assay)
10 nM-30 nM
RSK4
(Cell-free assay)
10 nM-30 nM
Src (T341M)
(Cell-free assay)
10 nM-30 nM
VEGFR2
(Cell-free assay)
10 nM-30 nM
In vitro

AT9283 leads to a clear polyploid phenotype by inhibiting the activity of Aurora B kinase in HCT116 cells with IC50 of 30 nM. Furthermore, this compound also produces the potent inhibition on HCT116 colony formation.

Kinase Assay
Aurora A and Aurora B Kinase Assays
Assays for Aurora A and B are performed in a DELFIA format. Aurora A enzyme is incubated with AT9283 and 3 μM cross-tide substrate (biotin-CGPKGPGRRGRRRTSSFAEG) in 10 mM MOPS, pH 7, 0.1 mg/mL BSA, 0.001% Brij-35, 0.5% glycerol, 0.2 mM EDTA, 10 mM MgCl2, 0.01% β-mercaptoethanol, 15 μM ATP, and 2.5% DMSO. Aurora B enzyme is incubated with this compound, 3 μM of the above substrate in 25 mM Tris, pH 8.5, 5 mM MgCl2, 0.1 mg/mL BSA, 0.025% Tween-20, 1 mM DTT, 15 μM ATP, and 2.5% DMSO. Reactions are allowed to proceed for 60 minutes and 45-90 minutes for Aurora A and Aurora B, respectively, before quenching with EDTA. The reaction mixtures are then transferred to a neutravidin-coated plate, and phosphorylated peptide is quantified by means of a phospho-specific antibody and a europium labeled secondary antibody using time-resolved fluorescence (excitation, 337 nm; emission, 620 nm). IC50 values for the control compounds are 92 nM (Aurora A assay) and 17 nM (Aurora B).
In vivo

In HCT116 human colon carcinoma xenograft bearing mice, AT9283 treatment (15 mg/kg and 20 mg/kg) for 16 days results in a significant tumor growth inhibition of 67% and 76%, respectively. In addition, this compound also exhibits a significantly longer half-life in tumors(2.5 hours) compared with plasma (0.5 hour) and modest oral bioavailability in mice (Fp.o. = 24%).

References

Applications

Methods Biomarkers Images PMID
Growth inhibition assay Cell viability
S1134-viability1
21430070

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT01145989 Completed
Multiple Myeloma
NCIC Clinical Trials Group|Astex Pharmaceuticals Inc.|Canadian Cancer Trials Group
 February 15 2011 Phase 2
NCT00443976 Completed
Non-Hodgkins Lymphoma|Unspecified Adult Solid Tumor Protocol Specific
NCIC Clinical Trials Group|Astex Pharmaceuticals Inc.|Canadian Cancer Trials Group
 January 30 2007 Phase 1

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