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

Validation & Quality Control

Customer Product Validation(4)

Quality Control & MSDS

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 2016-05-07)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02156895 Not yet recruiting Advanced Renal Cell Carcinoma Pfizer December 2016 --
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
NCT02693535 Recruiting Lymphoma, Non-Hodgkin|Multiple Myeloma|Advanced Solid Tumors American Society of Clinical Oncology|AstraZeneca|Bayer|B  ...more American Society of Clinical Oncology|AstraZeneca|Bayer|Bristol-Myers Squibb|Eli Lilly and Company|Genentech, Inc.|Pfizer 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 warmed (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.

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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

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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