Regorafenib (BAY 73-4506)

 

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Biological Activity

Description	Regorafenib (BAY 73-4506) is a multi-target inhibitor for VEGFR1, VEGFR2, VEGFR3, PDGFRβ, Kit, RET andRaf-1 with IC50 of 13 nM/4.2 nM/46 nM, 22 nM, 7 nM, 1.5 nM and 2.5 nM, respectively.
Targets	VEGFR1/2/3	PDGFRβ	Kit	RET	Raf-1
IC50	13 nM/4.2 nM/46 nM	22 nM	7 nM	1.5 nM	2.5 nM [1]
In vitro	Regorafenib strongly prevents VEGFR2 autophosphorylation in NIH-3T3/VEGFR2 cells with IC50 of 3 nM. In HAoSMCs, regorafenib suppress PDGFR-β autophosphorylation after stimulation with PDGF-BB, with an IC50 of 90 nM. Regorafenib also inhibits FGFR signaling in MCF-7 breast cancer (BC) cells stimulated with FGF10. Regorafenib very potently inhibited the mutant receptors KITK642E and RETC634W, with IC50 of approximately 20 nM and 10 nM, respectively. Regorafenib inhibits the proliferation of VEGF165-stimulated HUVECs, with an IC50 of approximately 3 nM. Regorafenib prevents the proliferation of FGF2-stimulated HUVECs and of PDGF-BB-stimulated HAoSMCs with IC50 of 127 nM and 146 nM, respectively. [1] Regorafenib targets both tumor cell proliferation and tumor vasculature through inhibition of receptors of tyrosine kinases (VEGFR, KIT, RET, FGFR, and PDGFR) and serine/threonine kinases (Raf and p38MAPK). [2] Regorafenib suppresses growth of human Hep3B, PLC/PRF/5 and HepG2 cells in a concentration- and time-dependent manner. [3]
In vivo	Regorafenib reveals potent dose-dependent TGI in various preclinical human xenograft models in mice, with tumor shrinkages in breast MDA-MB-231 and renal 786-O carcinoma models. Regorafenib prevents not only the growth of syngeneic primary 4T1 breast tumors growing orthotopically in the fat pad, but also suppresses the formation of tumor metastasis in the lung. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Kinase assays

In vitro assays using recombinant VEGFR2 (murine aa785–aa1367), VEGFR3 (murine aa818–aa1363), PDGFRβ (aa561–aa1106), Raf-1 (aa305–aa648) and BRafV600E (aa409–aa765) kinase domains are performed. Initial in vitro kinase inhibition profiling is performed at a fixed 1 μM Regorafenib concentration. Inhibitory concentration of 50% (IC50) values are determined from selected responding kinases, e.g., VEGFR1 and RET. TIE2 kinase inhibition is measured with a homogeneous time-resolved fluorescence (HTRF) assay using a recombinant fusion protein of glutathione-S-transferase, the intracellular domain of TIE2 and the peptide biotin-Ahx-EPKDDAYPLYSDFG as substrate.

Cell Assay: ^[1]

Cell lines	GIST 882 and TT cells
Concentrations	5 nM-10 μM
Incubation Time	96 hours
Method	For proliferation assays, GIST 882 and TT cells are grown in RPMI medium containing L-glutamine, and MDA-MB-231, HepG2 and A375 cells in DMEM always containing 10% hiFBS. Cells are trypsinized, plated at 5×104 cells/well in 96-well plates in complete media containing 10% FBS and grown overnight at 37 °C. The next day, vehicle or Regorafenib serially diluted in complete growth media to between 10 μM and 5 nM final concentrations, and 0.2% DMSO, is added and incubation is continued for 96 hours. Cell proliferation is quantified.

Animal Study: ^[1]

Animal Models	Female athymic NCr nu/nu mice with Colo-205, MDA-MB-231 or 786-O
Dosages	3 mg/kg, 10 mg/kg, 30 mg/kg, 100 mg/kg
Administration	Orally
Solubility	30% PEG400/0.5% Tween80/5% propylene glycol, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01959269	Recruiting	Colorectal Neoplasm	Bayer	2013-10
NCT01996969	Recruiting	Metastatic Colorectal Cancer	Seoul National University Hospital|Bayer	2013-10
NCT01973868	Recruiting	Neoplasms	Bayer	2013-11	Phase 1
NCT02001909	Not yet recruiting	Neoplasms	Bayer	2013-12	Phase 1
NCT01939223	Recruiting	Colorectal Neoplasms	Bayer|Onyx Pharmaceuticals	2013-12	Phase 3

Chemical Information

Download Regorafenib (BAY 73-4506) SDF

Chemical Name	1-(4-chloro-3-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.48282	4.8282	9.6564	14.4846

Product Citations (2)

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  Thyroid cancer cell resistance to gefitinib depends on the constitutive oncogenic activation of the ERK pathway [Frasca F,et al. J Clin Endocrinol Metab 2013;98(6):2502-12]

  
  PubMed: 23559083


• 
  

  Sorafenib/regorafenib and phosphatidyl inositol 3 kinase/thymoma viralproto-oncogene inhibition interact to kill tumor cells. [Sajithlal GB , et al. Mol Pharmacol 2013;84(4):562-71]

  
  PubMed: 23877009