High-throughput screening for compounds modulating expression of cellular macromolecules

Abstract

A method of screening for compounds that module expression of specific macromolecules, the “target”. The method is particularly useful in that it does not require separation of target-bound and excess ligand and therefore enables, but is not limited to, High Throughput Screening for compounds that increase or decrease the levels or amounts of a target present in a biological sample. The method can also be used for high-throughput diagnosis of a condition leading to an increase or decrease of a cellular macromolecule.

What is claimed is: 
     
         1 . A method of preventing or treating a neurological disease or disorder in an individual comprising: administering a therapeutically effective amount of one or more compounds comprising Astemizole, Tacrolimus, Lasalocid sodium, Monensin sodium, Emetine, Cetrimonium, analogs or combinations thereof. 
     
     
         2 . The method of  claim 1 , wherein the compounds are administered in a pharmaceutical composition. 
     
     
         3 . The method of  claim 1 , wherein a neurological disease or disorder comprises Creutzfeldt-Jakob disease; prion infection; Alzheimer's disease; Parkinson's disease, synucleinopathies; Multiple Sclerosis (MS); amyotrophic lateral sclerosis (ALS); Huntington's disease or neurological disorders associated with a virus infection. 
     
     
         4 . A method of modulating PrP in vitro or in vivo comprising, contacting a cell or administering to a subject an effective amount of one or more compounds comprising: Astemizole, Tacrolimus, Lasalocid sodium, Monensin sodium, Emetine, Cetrimonium, analogs or combinations thereof. 
     
     
         5 . The method of  claim 4 , wherein the amount of PrP is reduced as compared to a control. 
     
     
         6 . A method of modulating a protein kinase in vitro or in vivo, comprising contacting a cell or administering to a subject an effective amount of one or more compounds comprising: Astemizole, Tacrolimus, Lasalocid sodium, Monensin sodium, Emetine, Cetrimonium, analogs or combinations thereof. 
     
     
         7 . A method of preventing or treating cancer comprising: administering a therapeutically effective amount of one or more compounds comprising Astemizole, Tacrolimus, Lasalocid sodium, Monensin sodium, Emetine, Cetrimonium, analogs or combinations thereof. 
     
     
         8 . A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds comprising: Astemizole, Tacrolimus, Lasalocid sodium, Monensin sodium, Emetine, Cetrimonium, analogs or combinations thereof. 
     
     
         9 . A compound identified by method of screening for a candidate therapeutic compound comprising:
 screening a sample containing a specific target molecule in a high-throughput screening assay comprising the steps of: (i) contacting the sample with the candidate therapeutic compound (ii) adding a first and second ligand, wherein the first ligand comprises a first detectable label and the second ligand comprises a second detectable label, (iii) the first and second ligands each binding to separate and specific sites on a specific target molecule, wherein the screening assay does not require the step of (iv) washing;   detecting an emission of light when the first and second ligands specifically bind to the specific target molecule;   selecting the compound(s) that modulate(s) the amount of the target molecule as compared to a control; thereby, identifying the compound.   
     
     
         10 . A compound identified by a method comprising the steps of:
 placing the sample containing a specific target molecule into a receptacle permitting irradiation of the sample at a wavelength suitable for exciting the donor fluorophore and measurement of the fluorescence of the acceptor fluorophore via a high-throughput assay;   contacting the sample with the candidate therapeutic compound;   adding a first ligand that binds to a specific site on the target molecule wherein the first ligand is linked to a first fluorophore (the “donor fluorophore”);   adding a second ligand that binds to a specific site on the same target molecule distinct from that to which the first ligand binds wherein the second ligand is linked to a second fluorophore (the “acceptor fluorophore”);   irradiating the sample containing the target molecule linked to the ligands at a wavelength optimal for exciting the donor fluorophore and measuring the intensity of the light emitted by the acceptor fluorophore;   selecting the compound(s) that modulate(s) the amount of the target molecule as compared to a control; thereby,
 screening for a candidate compound. 
   
     
     
         11 . The method of  claim 9  or  10 , wherein the compound modulates an amount, a function, activity or expression of a target molecule as measured by the emission of light. 
     
     
         12 . A pharmaceutical composition comprising a compound identified by the method of  claim 9  or  10 .