Ridaforolimus (Deforolimus, MK-8669)

 

 

Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

 

Biological Activity

Description	Ridaforolimus (Deforolimus) is a selective mTOR inhibitor with IC50 of 0.2 nM; while not classified as a prodrug, mTOR inhibition and FKBP12 binding is similar to rapamycin.
Targets	mTOR
IC50	0.2 nM [1]
In vitro	Treatment of HT-1080 cells with Deforolimus induces a dose-dependent inhibition of phosphorylation of both S6 and 4E-BP1, with IC50 of 0.2 nM and 5.6 nM, respectively, and leads to a decrease in cell size, an increase in the proportion of cells in the G1 phase of the cell cycle, and inhibition of glucose uptake. Deforolimus displays significant antiproliferative activity a broad panel of cell lines with EC50 of 0.2-2.3 nM. Deforolimus potently and selectively inhibits VEGF production in a dose-dependent manner. [1] Deforolimus treatment significantly induces growth suppression in human NSCLC cell lines with IC30 values of 2.45-8.83 nM, with the exception of H157 with IC30 of >20 nM. Deforolimus treatment (2.8-5.9 nM) significantly dephosphorylates p70S6KThr389 in A549, H1703 and H157 cells, except H1666 that may express a resistant variant of mTORC1, and causes increased phosphorylation of pAKTser473 and pAKTThr308 in A549 and H1703 cells. Deforolimus in combination with the MEK inhibitors, CI-1040 or PD0325901 exhibits dose-dependent synergism in lung cancer cell lines, which is associated with the suppression of proliferation rather than enhancement of cell death, involving the inhibition of ribosomal biogenesis by 40% within 24 hours and a decreased polysome/monosome ratio. [2]
In vivo	Administration of Deforolimus exerts significant antitumor effects in mice bearing PC-3 (prostate), HCT-116 (colon), MCF7 (breast), PANC-1 (pancreas) or A549 (lung) xenografts in a dose-dependent manner, and inhibits mTOR signaling in in SK-LMS-1 xenograft model associated with inhibition of tumor growth. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cell based target inhibition

HT-1080 cells are treated with increasing concentrations of Deforolimus (0-100 nM) for 2 hours, prior to harvest. Cellular lysates are extracted in denaturing lysis buffer, resolved on SDS-PAGE and transferred to PVDF membranes. After blocking, membranes are incubated with primary antibodies for 1 hour, followed by appropriate HRPconjugated secondary antibodies for 1 hour at room temperature. Immunoreactive proteins are detected using enhanced chemiluminescence and autoradiography performed by exposure to X-ray film. IC50 is determined from the inhibition of levels of phosphorylated ribosomal protein S6 (p-S6) and 4E-BP1 (p-4E-BP1).

Cell Assay: ^[2]

Cell lines	Colo205, H1755, H1395, H1666, A549, H157, and H1703 cells
Concentrations	Dissolved in ethanol, final concentrations ~ 1 μM
Incubation Time	72-120 hours
Method	Cells are seeded at 2-3 × 104/mL, and serial dilutions of Deforolimus are added after 2 hours, for at least three cell doublings (72-120 hours). Deforolimus effects are measured using the CellTiter 96 Aqueous nonradioactive cell proliferation assay and Sulforhodamine B assays. For Deforolimus, growth effects are described as IC30 because rapamycin and its derivatives do not significantly impede cell proliferation.

Animal Study: ^[1]

Animal Models	Male and female athymic NCr-nu mice with xenografts established by subcutaneous implantation of PC-3, A549, HCT-116, MCF7, PANC-1 and SK-LMS-1 tumors
Dosages	~10 mg/kg
Administration	Intraperitoneally
Solubility	30% PEG400/0.5% Tween80/5% propylene glycol, 20 mg/mL

1

References

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

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01256268	Recruiting	Endometrial Cancer|Ovarian Cancer	H. Lee Moffitt Cancer Center and Research Institute|Merck	2011-06	Phase 1
NCT01380184	Completed	Cancer, Advanced	Merck	2011-07	Phase 1
NCT01431534	Active, not recruiting	Solid Tumors	Merck	2012-01	Phase 1
NCT01431547	Active, not recruiting	Solid Tumors	Merck	2012-02	Phase 1
NCT01605396	Active, not recruiting	Breast Neoplasms	Merck	2012-07	Phase 2

Chemical Information

Download Ridaforolimus (Deforolimus, MK-8669) SDF

Chemical Name

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.99021	9.9021	19.8042	29.7063

Customer Reviews (1)

Click to enlarge	Rating
	Source	Dr. Zhang of Tianjin Medical University. Ridaforolimus (Deforolimus, MK-8669) purchased from Selleck
	Method	Western blotting
	Cell Lines	Breast cancer cells
	Concentrations	0-5 nM
	Incubation Time	24 h
	Results	Deforolimus treatment resulted in a reduction of mTOR phosphorylation in Breast cancer cells.

Product Citations (2)

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  Inhibition of glycogen biosynthesis via mTORC1 suppression as an adjunct therapy for Pompe disease. [Ashe KM, et al. Mol Genet Meta 2010;100(4):309-15]

  
  PubMed: 20554235


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  Activation of mammalian target of rapamycin contributes to pain nociception induced in rats by BmK I, a sodium channel-specific modulator. [Jiang F, et al. Neurosci Bull 2013;10.1007/s12264-013-1377-0]

  
  PubMed: 24132796