Back to results
US 10,274,481 B2Granted
Method for identifying modulators of notch signaling
AssigneeEcole Polytechnique Federale De Lausanne (Epfl)
InventorsFreddy Radtke, Rajwinder Lehal, Viktoria Reinmüller, Jieping Zhu
Life of this patent
Estimating the term…
Abstract
The present invention relates to use of inhibitors of Notch signalling pathway selected from the group consisting of 6-(4-Tert-Butylphenoxy)Pyridin-3-Amine (I3), its derivatives, in treating and/or preventing cancers.
Claims25
A method for identifying modulators of NOTCH signalling in a cell culture, comprising
- (i) co-culturing mammalian cells expressing a NOTCH ligand and mammalian cells expressing a NOTCH receptor to form a co-culture, wherein said mammalian cells expressing a NOTCH ligand are HeLa cells that have been transfected with a NOTCH ligand expression system and said mammalian cells expressing a NOTCH receptor are HeLa cells that have been transfected with a NOTCH receptor expression system;
- (ii) incubating the co-culture with and without a candidate modulator of NOTCH signalling to be tested
- (iii) quantifying the amount of a NOTCH intracellular domain (NICD) that has been released from a NOTCH receptor;
- and/or
- (iv) quantifying the NICD-mediated modulation of a NOTCH signalling specific reporter system, wherein a reduction in the signal of the NOTCH signalling specific reporter system, a reduction of the amount of NICD, the absence of NICD and/or the absence of the signal of the NOTCH signalling specific reporter system in said co-culture that has been incubated with a candidate modulator of NOTCH signalling, compared to the signal of the NOTCH signalling specific reporter system and/or the amount of NICD in said co-culture that has been incubated without a candidate modulator of NOTCH signalling, is indicative of NOTCH signalling modulation.
2 more elements
A method for identifying modulators of NOTCH signalling by high throughput screening of a library comprising candidate modulators of NOTCH signalling comprising the steps of:
- (a) providing an assay in wells of a microwell plate wherein the assay comprises a co-culture comprising mammalian cells expressing a NOTCH ligand and mammalian cells expressing a NOTCH receptor, wherein said mammalian cells expressing a NOTCH ligand are HeLa cells that have been transfected with a NOTCH ligand expression system and said mammalian cells expressing a NOTCH receptor are HeLa cells that have been transfected with a NOTCH receptor expression system;
- (b) incubating the co-culture of said assay with and without a candidate modulator of NOTCH signalling to be tested;
- and
- (c)(i) quantifying the amount of a NOTCH intracellular domain (NICD) that has been released from the NOTCH receptor;
- and/or
- (c)(ii) quantifying the NICD-mediated modulation of a NOTCH signalling specific reporter system;
- wherein a reduction in the signal of the NOTCH signalling specific reporter system, a reduction of the amount of NICD, the absence of NICD and/or the absence of the signal of the NOTCH signalling specific reporter system in said co-culture that has been incubated with a candidate modulator of NOTCH signalling, compared to the signal of the NOTCH signalling specific reporter system and/or the amount of NICD in said co-culture that has been incubated without a candidate modulator of NOTCH signalling, is indicative of NOTCH signalling modulation.
4 more elements
Dependencies are inferred from the claim text.