Vemurafenib (PLX4032)

Biological Activity

Vemurafenib (PLX4032) is the first and effective selective inhibitor of B-Raf, which can inhibit RAFV600E and C-RAF-1 activity with IC50 of 31 nM and 48 nM, respectively. Vemurafenib induces autophagy.

Product Citations

• 

  Oncogene. 2021 Jun;40(24):4137-4150.

  Targeting the p300/NONO axis sensitizes melanoma cells to BRAF inhibitors
  Vemurafenib (PLX4032) purchased from AbMole

• 

  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
  Vemurafenib (PLX4032) purchased from AbMole

• 

  J Nucl Med. 2018 Nov 21.

  MEK inhibition induces therapeutic iodine uptake in a murine model of anaplastic thyroid cancer.
  Vemurafenib (PLX4032) purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Cell Death Differ (2016). Figure 2. PLX4032
	Method	western blot
	Cell Lines	BRAF mutant melanoma cell lines
	Concentrations	3 μM
	Incubation Time	24 h
	Results	PLX4032-induced upregulation of BIM and PUMA occurred normally in the control sgRNA transduced cell lines.

	Source	Cell Death Differ (2016). Figure 1. PLX4032
	Method	flow cytometry
	Cell Lines	BRAFWT cells
	Concentrations	0.01, 0.03, 0.1, 0.3, 1, 3, 10 μM
	Incubation Time	24 h
	Results	We found that PLX4032 induced significant dose-dependent apoptosis over a time course of 5 days in all BRAF mutant melanoma cell lines (M14, UACC257, Malme3M, SKMEL5 and UACC62), whereas the BRAFWT cells (CHL-1) were unaffected.

Protocol (for reference only)

Cell Experiment
Cell lines	MDA-MB-435 cells line
Preparation method	Cellular proliferation assays Cellular proliferation was evaluated by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT; Sigma) assay. Briefly, cells were plated in 96-well microtiter plates at a density of 1,000 to 5,000 cells per well in a volume of 180 μL. For the assay, RG7204 was prepared at 10 times the final assay concentration in media containing 1% DMSO. Twenty-four hours after cell plating, 20 μL of the appropriate dilution were added to plates in duplicate. The plates were assayed for proliferation 6 days after the cells were plated according to the procedure originally described by Mosmann . The IC50 was determined from the regression of a plot of the logarithm of the concentration versus percent inhibition by XLfit (version 4.2; IDBS) using the Dose-Response One-Site Model (#205).
Concentrations	0~10µM
Incubation time	6 days

Animal Experiment
Animal models	Colo829 and A375 xenografts
Formulation	an aqueous vehicle containing 2% Klucel LF (Hydroxypropylcellulose; Aqualon) and adjusted to pH 4 with dilute HCl.
Dosages	100 mg/kg bid
Administration	oral gavage

Chemical Information

Molecular Weight	489.92
Formula	C23H18ClF2N3O3S
CAS Number	918504-65-1
Solubility (25°C)	DMSO 60 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] Shao Y et al. Cell Death Differ. BH3-only protein silencing contributes to acquired resistance to PLX4720 in human melanoma.

[2] Patrawala S et al. Future Oncol. Vemurafenib (RG67204, PLX4032): a potent, selective BRAF kinase inhibitor.