印度威罗菲尼

产品描述

Vemurafenib (PLX4032) is a novel and potent B-Raf (V600E) inhibitor (IC50: 31 nM).

靶点活性

ACK1,19 nM (cell free)

BLK,547nM

B-Raf,100nM

B-Raf (V600E),31 nM (cell free)

BRK,213nM

C-Raf,48nM

CSK,2.339μM

FGR,63nM

FRK (PTK5),1.884μM

Lck,183nM

Lyn B,599nM

MAP4K5 (KHS1),51nM

MNK2,1.717μM

NEK11,317nM

Src,2.389μM

SRMS,18 nM (cell free)

WNK3,877nM

Yes1,604nM

c-Raf-1,48 nM (cell free)

实验溶液

30% PEG400+0.5% Tween80+5% propylene glycol: 5 mg/mL

体外活性

Vemurafenib (PLX4032, RG7204) displays similar potency for B-RAFV600E (31nM) and c-RAF-1 (48nM) and selectivity against many other kinases, including wild type B-RAF (100nM) [1]. In 17 melanoma cell lines, RG7204 was a potent inhibitor of proliferation in those expressing BRAFV600E but not BRAFWT. RG7204 also potently inhibited proliferation of melanoma cell lines expressing other codons 600 BRAF mutations (V600D, V600K, and V600R) [2]. Melanoma cells were more sensitive to PLX4032 than CRC cells. The inhibition of EGFR does not significantly affect the proliferation of EGFR in WiDr cells. In contrast, suppression of EGFR in combination with PLX4032 caused a marked inhibition of proliferation in WiDr cells [3].

体内活性

In BRAFV600E-mutant xenograft, PLX4032 (6 or 8 mg/kg) demonstrated dose-dependent inhibition of tumor growth, with higher exposures resulting in tumor regression [1]. In mice bearing Colo829 tumor xenografts, RG7204 at 100 mg/kg bid for 21 days showed greatly improved antitumor activity at the end of the study on day 38 after the tumor cell implant. There was complete tumor regression in all 10 mice treated with RG7204 by the end of the study [2]. In the dose-escalation cohort, among the 16 patients with melanoma whose tumors carried the V600E BRAF mutation and who were receiving 240 mg or more of PLX4032 twice daily, 10 had a partial response and 1 had a complete response [4].

激酶实验

Expression and purification of B-RAF, structure determination, and protein kinase activity measurements were carried out as previously described. To obtain co-crystals of B-RAFV600E with PLX4032, the protein solution was initially mixed with the compound dissolved in DMSO at a final compound concentration of 1 mM. This complex was co-crystallized by a sitting drop vapor diffusion experiment in which equal volumes of complex (at 10 mg/ml concentration) and reservoir solution (100mM BisTris at pH 6.0, 12.5% 2,5-hexanediol, and 12% PEG3350) were mixed and allowed to equilibrate against the reservoir at 4°C. The crystal was soaked in cryosolvent, followed by flash-freezing in liquid nitrogen. The data were collected at Beamline ALS831 with the wavelength of 1.11?. The Ramachandran plot from the refined structure shows that 94%, 5.6% and 0.4% residues are in the most favored, additional allowed and generously allowed regions, respectively. A summary of the crystallography statistics is included in Supplementary Table 3. COLO205 tumor xenograft studies (Molecular Imaging Research, Ann Arbor, MI) were carried out as previously described either using a conventional formulation (5%DMSO, 1% methylcellulose) or using the MBP formulation [1].

细胞实验

Cellular proliferation was evaluated by MTT 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 [2].

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

动物实验

All animal procedures were approved by the Ethical Commission of the Institute for Cancer Research and Treatment and by the Italian Ministry of Health. WiDr cells were injected subcutaneously into the right posterior flanks of 7-week-old immunodeficient NODSCID female mice (6 mice per group). Tumour formation was monitored twice a week, and tumour volume based on caliper measurements was calculated by the modified ellipsoidal formula: tumour volume = 1/2 length × width. When tumours reached a volume of approximately 200–250 mm^3, mice were randomly assigned to treatment with vehicle or drug(s) [3].

动物模型：Mice (athymic nude) xenograft models of LOX, Colo829, and A375 cells

分子量

489.92

分子式

C23H18ClF2N3O3S

CAS

918504-65-1

溶解度

DMSO: 90 mg/mL (183.7 mM)

Ethanol: <1 mg/mL

Water: <1 mg/mL

（ < 1 mg/ml refers to the product slightly soluble or insoluble )

储存条件

store at -80°C

备注

For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.