Axitinib Licensed by Pfizer

Catalog No.S1005

Axitinib is a multi-target inhibitor of VEGFR1, VEGFR2, VEGFR3, PDGFRβ and c-Kit with IC50 of 0.1 nM, 0.2 nM, 0.1-0.3 nM, 1.6 nM and 1.7 nM in Porcine aorta endothelial cells, respectively.

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Axitinib Chemical Structure

Axitinib Chemical Structure
Molecular Weight: 386.47

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Related Compound Libraries

VEGFR Inhibitors with Unique Features

Product Information

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  • Research Area
  • Axitinib Mechanism
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Axitinib is a multi-target inhibitor of VEGFR1, VEGFR2, VEGFR3, PDGFRβ and c-Kit with IC50 of 0.1 nM, 0.2 nM, 0.1-0.3 nM, 1.6 nM and 1.7 nM in Porcine aorta endothelial cells, respectively.
Targets VEGFR1/FLT1 [1]
(Porcine aorta endothelial cells)
VEGFR2/Flk1 [1]
(Porcine aorta endothelial cells)
VEGFR2/KDR [1]
(Porcine aorta endothelial cells)
VEGFR3 [1]
(Porcine aorta endothelial cells)

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IC50 0.1 nM 0.18 nM 0.2 nM 0.1 nM-0.3 nM
In vitro Axitinib could block the cellular autophosphorylation of VEGFR and VEGF-mediated endothelial cell viability, tube formation, and downstream signaling. Axitinib inhibits the proliferation of variable cell lines with IC50 of >10,000 nM (IGR-N91), 849 nM (IGR-NB8), 274 nM (SH-SY5Y) and 573 nM (non-VEGF stimulated HUVEC). [2]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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... Click to View More Cell Line Experimental Data

In vivo Axitinib exhibits primary inhibition to orthotopically transplanted models such as M24met (melanoma), HCT-116 (colorectal cancer), and SN12C (renal cell carcinoma). [1] Axitinib delays the tumor growth with 11.4 days compared to the controls (p.o. 30 mg/kg) and decreases the Mean Vessels Density (MVD) to 21, compared to 49 in controls, in IGR-N91 flank xenografts. [2] Axitinib significantly inhibits growth and disrupts tumor microvasculature in BT474 breast cancer model at 10-100 mg/kg. [3] Axitinib has shown single-agent activity in variable tumors, including renal cell carcinoma, thyroid cancer, non-small cell lung cancer, and melanoma.
Features Superior as second-line therapy relative to sorafenib (current standard-of-care).

Protocol(Only for Reference)

Kinase Assay:

[1]

Cellular receptor kinase phosphorylation assay Porcine aorta endothelial (PAE) cells, which overexpress full-length VEGFR2, PDGFRβ, Kit, and NIH-3T3, which overexpress murine VEGFR2 (Flk-1) or PDGFRα, are generated. The 96-well plates are coated with 100 μL/well of 2.5 μg/mL anti-VEGFR2 antibody, 0.75 μg/mL anti-PDGFRβ antibody, 0.25 μg/mL anti-PDGFRα antibody, 0.5 μg/mL anti-KIT antibody, or 1.20 μg/mL anti-Flk-1 antibody to prepare ELISA capture plates. Then phosphorylation of RTK is measured by ELISA.

Cell Assay:

[2]

Cell lines HUVEC, SH-SY5Y, IGR-N91 and IGR-NB8 cells
Concentrations 1 nM - 10 μM
Incubation Time 72 hours
Method

Cells are seeded in a 96-well plate at a density of 5 × 104 and cultured for 24 hours. Axitinib is added to the cells at concentrations ranging from 1 nM to 10 μM. Cell viability is measured after 72 hours by MTS tetrazolium substrate and IC50 values are calculated.

Animal Study:

[3]

Animal Models BT474 breast cancer cells are implanted subcutaneously into Immune-deficient female mice (Nu/nu; age 8-12 weeks).
Formulation 0.5% carboxymethylcellulose (CMC)
Dosages 10, 30 or 100 mg/kg
Administration Oral daily

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Hu-Lowe DD, et al. Clin Cancer Res, 2008, 14(22), 7272-7283.

[2] Rossler, J. et al., Int J Cancer, 2011, 128(11), 2748-2758.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-07-23)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02156895 Not yet recruiting Advanced Renal Cell Carcinoma Pfizer December 2016 --
NCT02782403 Not yet recruiting Leukemia M.D. Anderson Cancer Center|Pfizer August 2016 Phase 1|Phase 2
NCT02762513 Not yet recruiting Squamous Cell Carcinoma of the Head and Neck University of Michigan Cancer Center May 2016 Phase 2
NCT02684006 Recruiting Renal Cell Cancer Pfizer March 2016 Phase 3
NCT02636725 Recruiting Alveolar Soft Part Sarcoma|Soft Tissue Sarcomas Breelyn Wilky|Merck Sharp & Dohme Corp.|Pfizer|University  ...more Breelyn Wilky|Merck Sharp & Dohme Corp.|Pfizer|University of Miami March 2016 Phase 2

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Chemical Information

Download Axitinib SDF
Molecular Weight (MW) 386.47
Formula

C22H18N4OS

CAS No. 319460-85-0
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms AG 013736
Solubility (25°C) * In vitro DMSO 35 mg/mL warming (90.56 mM)
Water <1 mg/mL (<1 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 0.5% CMC 30 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name N-methyl-2-[[3-[(1E)-2-(2-pyridinyl)ethenyl]-1H-indazol-6-yl]thio]-benzamide

Customer Product Validation(4)


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Rating
Source J Biomol Screen, 2011,16: 141-154. Axitinib purchased from Selleck
Method Spheroid Staining & Sectioning
Cell Lines MDA-MB-231 cells
Concentrations 10 µM
Incubation Time 72 h
Results These results demonstrate that both U0126 and axitinib penetrated into the spheroid, resulting in downregulation of vimentin throughout the whole spheroid by more than 80% as compared to untreated spheroids.

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Rating
Source J Biomol Screen, 2011,16: 141-154. Axitinib purchased from Selleck
Method Western Blot
Cell Lines MDA-MB-231 cells
Concentrations 10 µM
Incubation Time
Results Based on these data, there appears to be a threshold of 50% downregulation of VimPro-Fluc activity that must be reached before protein downregulation occurs.

Click to enlarge
Rating
Source J Biomol Screen 2011 16, 141-154. Axitinib purchased from Selleck
Method Secondary assay
Cell Lines MDA-MB-231 cells
Concentrations 10 µM
Incubation Time
Results U0126, pF2341066, axitinib, and pKC412 caused significant inhibition of the invasive potential of Mda-Mb-231 spheroids. Conversely, dasatinib, a potent inhibitor of vimentin gene expression, did not significantly alter the invasive potential of MDA-MB-231 spheroids.

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Rating
Source Dr. Cheri Pasch of UW Madison. Axitinib purchased from Selleck
Method Western blot
Cell Lines HUVEC cells
Concentrations 0.1-10 nM
Incubation Time
Results Axitinib treatment blocks VEGFR2 phosphorylation.

Tech Support

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