Vemurafenib (PLX4032)

Catalog No.S1267 Batch:S126712

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Technical Data

Formula

C23H18ClF2N3O3S
Molecular Weight 489.92 CAS No. 918504-65-1
Solubility (25°C)* In vitro DMSO 97 mg/mL (197.99 mM)
Water Insoluble
Ethanol Insoluble
* <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.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description Vemurafenib (PLX4032, RG7204, RO5185426) is a novel and potent inhibitor of B-RafV600E with IC50 of 31 nM in cell-free assay. 10-fold selective for B-RafV600E over wild-type B-Raf in enzymatic assays and the cellular selectivity can exceed 100-fold. Vemurafenib (PLX4032, RG7204) induces autophagy.
Targets
SRMS [1]
(Cell-free assay)		 ACK1 [1]
(Cell-free assay)		 B-Raf (V600E) [1]
(Cell-free assay)		 C-Raf [1]
(Cell-free assay)		 MAP4K5 (KHS1) [1]
(Cell-free assay)		 View More
18 nM 19 nM 31 nM 48 nM 51 nM
In vitro PLX4032 inhibits B-RAFV600E, C-RAF, as well as wildtype B-RAF, with IC50 of 31 nM, 48 nM and 100 nM, respectively. PLX4032 also inhibits several non-RAF kinases, including ACK1, KHS1, and SRMS, with IC50 of 18 nM to 51 nM. [1] In melanoma cell lines, the inhibitory effect by PLX4032 depends on B-RAF mutational status, because PLX4032 potently inhibits those harboring B-RAF V600 mutants, including V600E, V600D, V600K, and V600R, but not wildtype or other mutants. The IC50 values of PLX4032 on these cells, including MALME-3M, Colo829, Colo38, A375, SK-MEL28, and A2058, ranges from 20 nM to 1 μM. In these cells, PLX4032 (0.1 μM to 30 μM) also inhibits the phosphorylation of both MEK1/2 and ERK1/2. [2] PLX4032 is highly effective in the treatment of melanoma, for its ability of inhibiting B-RAFV600E. However, PLX4032 displays limited effect in colon cancer patients that also carrying B-RAFV600E oncoprotein. The reason for this is that, in colon cancer cells, B-RAFV600E inhibition by PLX4032 results in a rapid feedback EGFR activation, which compensates for the PLX4032-inhibited cell proliferation. [3]
In vivo In B-RAFV600E-mutant mice xenograft models, PLX4032 (6 mg/kg–20 mg/kg) inhibits tumor growth. [1] In mice xenograft models of LOX, Colo829, and A375 cells, PLX4032 (12.5 mg/kg–100 mg/kg) inhibits tumor growth and prolongs mice survival. [2]
Features A novel and potent inhibitor of the B-RAFV600E oncoprotein.

Protocol (from reference)

Kinase Assay:

[1]
  • RAF kinase activity measurements

    The kinase activities of wild-type RAF and mutants are determined by measuring phosphorylation of biotinylated-BAD protein. For each enzyme (0.01 ng), 20 μL reactions are carried out in 20 mM Hepes (pH 7.0), 10 mM MgCl2, 1 mM DTT, 0.01% (v/v) Tween-20, 50 nM biotin-BAD protein, and 1 mM ATP at room temperature. Reactions are stopped at 5 min with 5 μL of a solution containing 20 mM Hepes (pH 7.0), 200 mM NaCl, 80 mM EDTA, 0.3% (w/v) bovine serum albumin (BSA). The stop solution also includes phospho-BAD (Ser112) antibody, streptavidin-coated donor beads, and protein A acceptor beads. The antibody and beads are pre-incubated in stop solution in the dark at room temperature for 30 min. The final dilution of antibody is 1/2000 and the final concentration of each bead is 10 μg/mL. The assay plates are incubated at room temperature for one hour and then are read on a PerkinElmer AlphaQuest reader. Mutant activities are the average of two different batches of purified protein assayed in duplicate in three different experiments.
Cell Assay:

[2]
  • Cell lines

    MALME-3M, Colo829, Colo38, A375, SK-MEL28, and A2058 cells

  • Concentrations

    0–10 μM , dissolved in DMSO

  • Incubation Time

    5 days

  • Method

    Cellular proliferation is evaluated by MTT assay. Briefly, cells are plated in 96-well microtiter plates at a density of 1000 to 5000 cells per well in a volume of 180 μL. PLX4032 is 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 of PLX4032 are added to plates in duplicate. The plates are assayed for proliferation 6 days after the cells are plated. Percent inhibition is calculated and the IC50 is determined from the regression of a plot of the logarithm of the concentration versus percent inhibition.
Animal Study:

[2]
  • Animal Models

    Mice (athymic nude) xenograft models of LOX, Colo829, and A375 cells

  • Dosages

    12.5 mg/kg–100 mg/kg

  • Administration

    Oral gavage twice daily

Customer Product Validation

Data from [Data independently produced by Nature, 2015, 520(7547), 368-72]

Data from [Data independently produced by Nature, 2015, 520(7547), 368-72]

Data from [Data independently produced by Oncogene, 2015, 10.1038/onc.2015.97]

Data from [Data independently produced by Nature, 2014, 508(7494), 118-22]

Selleck's Vemurafenib (PLX4032) has been cited by 749 publications

The ribotoxic stress response drives UV-mediated cell death [ Cell, 2024, 187(14):3652-3670.e40] PubMed: 38843833
Proteostatic reactivation of the developmental transcription factor TBX3 drives BRAF/MAPK-mediated tumorigenesis [ Nat Commun, 2024, 15(1):4108] PubMed: 38750011
Proteostatic reactivation of the developmental transcription factor TBX3 drives BRAF/MAPK-mediated tumorigenesis [ Nat Commun, 2024, 15(1):4108] PubMed: 38750011
Mi-2β promotes immune evasion in melanoma by activating EZH2 methylation [ Nat Commun, 2024, 15(1):2163] PubMed: 38461299
The CoREST repressor complex mediates phenotype switching and therapy resistance in melanoma [ J Clin Invest, 2024, 134(6)e171063] PubMed: 38300709
Ubiquitin-specific protease 22 controls melanoma metastasis and vulnerability to ferroptosis through targeting SIRT1/PTEN/PI3K signaling [ MedComm (2020), 2024, 5(8):e684] PubMed: 39135915
Genomic deletions explain the generation of alternative BRAF isoforms conferring resistance to MAPK inhibitors in melanoma [ Cell Rep, 2024, S2211-1247(24)00376-0] PubMed: 38614086
ERK1/2 interaction with DHPS regulates eIF5A deoxyhypusination independently of ERK kinase activity [ Cell Rep, 2024, 43(10):114831] PubMed: 39392755
Cell-specific models reveal conformation-specific RAF inhibitor combinations that synergistically inhibit ERK signaling in pancreatic cancer cells [ Cell Rep, 2024, 43(9):114710] PubMed: 39240715
ZEB1 controls a lineage-specific transcriptional program essential for melanoma cell state transitions [ Oncogene, 2024, 43(20):1489-1505] PubMed: 38519642

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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