Axitinib

Biological Activity

Axitinib blocked phosphorylation of VEGFR-2 and VEGFR-3 with average IC50s of 0.2 and 0.1 to 0.3 nM.Axitinib (also known as AG013736) is a small molecule tyrosine kinase inhibitor.It inhibits multiple targets, including VEGFR-1, VEGFR-2, VEGFR-3, platelet derived growth factor receptor (PDGFR), and cKIT (CD117). It has been shown to significantly inhibit growth of breast cancer in xenograft models and has been successful in trials with renal cell carcinoma (RCC) and several other tumor types.

Product Citations

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  J Virol. 2020 Feb 14;94(5):e01791-19.

  Targeting Kaposi's Sarcoma-Associated Herpesvirus ORF21 Tyrosine Kinase and Viral Lytic Reactivation by Tyrosine Kinase Inhibitors Approved for Clinical Use
  Axitinib purchased from AbMole

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  Oncol Res. 2019 Sep 23;27(9):987-995.

  Nutlin-3-Induced Sensitization of Non-Small Cell Lung Cancer Stem Cells to Axitinib-Induced Apoptosis Through Repression of Akt1/Wnt Signaling.
  Axitinib purchased from AbMole

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  Int J Cancer. 2017 May 1;140(9):2150-2161.

  Complementary actions of dopamine D2 receptor agonist and anti-vegf therapy on tumoral vessel normalization in a transgenic mouse model
  Axitinib purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Int J Cancer (2017) . Figure 6. Axitinib (20 mg/kg, Abmole Bioscience)
	Method
	Cell Lines	Hmga2/T mice
	Concentrations	20mg/kg
	Incubation Time	6 weeks
	Results	Together, these results show that, while bromocriptine and/or axitinib were able to correct structuralabnormalities of tumoral vessels with a similar efficacy, only the combination treatment restored their function.

	Source	Int J Cancer (2017) . Figure 5. Axitinib (20 mg/kg, Abmole Bioscience)
	Method
	Cell Lines	Hmga2/T mice
	Concentrations	20mg/kg
	Incubation Time	6 weeks
	Results	Analysis of pituitary vasculature in various conditions and morphometric measurements of blood vessels demonstrate that D2R agonist or axitinib had an equivalent capacity to improve structural defects present in tumoral blood vessels and that the combination treatment did not lead to a significant advantage.

	Source	Int J Cancer (2017) . Figure 4. Axitinib (20 mg/kg, Abmole Bioscience)
	Method
	Cell Lines	Hmga2/T mice
	Concentrations	20mg/kg
	Incubation Time	6 weeks
	Results	Importantly, the expression of Angpt1 and Prok1, which was upregulated in response to bromocriptine treatment, was unchanged and remained low after axitinib treatment (Fig. 4d).

Protocol (for reference only)

Cell Experiment
Cell lines	HUVEC, SH-SY5Y, IGR-N91 and IGR-NB8 cell lines
Preparation method	MTS cell proliferation assay HUVEC, SH-SY5Y, IGR-N91 and IGR-NB8 cell lines were seeded at 5,000 cells per well in a 96-well plate and left to settle overnight in EMBV R-2 or DMEM with 10% fetal calf serum. The cells were treated with axitinib at concentrations ranging from 1 nmol/l to 10 lmol/l. Cell viability was determined after 72 hr using MTS tetrazolium substrate (CellTiter 96 Aqueous One Solution Cell Proliferation Assay; Promega Corporation, Charbonnieres, France) and colorimetric measurement at 490 nm in an automatic plate reader (Elx808; Fisher Bioblock Scientific SAS, Illkirch, France). IC50 values were calculated using GraphPadV R Prism software (version 3.00).
Concentrations	1 nM - 10 μM
Incubation time	72 hr

Animal Experiment
Animal models	IGR-N91 xenografts with female SPF-Swiss athymic nude mice at 6–8 weeks of age
Formulation	0.5% CMC
Dosages	30 mg/kg BID for 2 weeks
Administration	oral gavage

Chemical Information

Molecular Weight	386.47
Formula	C22H18N4OS
CAS Number	319460-85-0
Solubility (25°C)	DMSO ≥ 20 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Carmichael et al. Clin Adv Hematol Oncol. Comprehensive overview of axitinib development in solid malignancies: focus on metastatic renal cell carcinoma.

[2] Pithavala et al. Cancer Chemother Pharmacol. Evaluation of the effect of food on the pharmacokinetics of axitinib in healthy volunteers.

[3] Jochen Rössler, et al. Int J Cancer. The selective VEGFR1-3 inhibitor axitinib (AG-013736) shows antitumor activity in human neuroblastoma xenografts