Odyssey Thera Inc.

Signal Pharmaceuticals, Inc.

John K Westwick

Marnie L MacDonald

Stephen William Watson Michnick

Ingrid Remy

Helen Yu

Jane Lamerdin

Bernd M Stein

Brigitte Keon

Bruce W Ennis

Marnie Macdonald

The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. Single-color and multi-color assays are disclosed. Further disclosed are universal expression vectors with cassettes that allow the rapid construction of assays for a large and diverse number of gene/reporter combinations. The development of such assays is shown to be straightforward, providing for a broad, flexible and biologically relevant platform for drug discovery.

Protein fragment complementation assays for high-throughput and high-content screening

The invention provides a method of screening a candidate drug, a compound library or a biological extract to identify activators or inhibitors of G-protein-coupled receptors or G-protein-coupled pathways, comprising: (A) using a fluorescent protein fragment complementation assay to construct an assay for one or more steps in a G-protein-coupled pathway; (B) testing the effects of the candidate drugs, compound library, or biological extract on the receptor or pathway of interest; and (C) using the results of the screening to identify specific agents that activate or inhibit the receptor or pathway of interest. The invention also provides a method for identifying a drug lead that modulates the activity of a G-protein-coupled pathway using a fluorescent protein fragment complementation assay. The method of the invention is used to identify agonists, antagonists, activators or inhibitors of G-protein coupled receptors or G-protein-coupled pathways.

Fragment complementation assays for G-protein-coupled receptors and their signaling pathways

The present invention relates to methods and compositions designed for the treatment, management or prevention of cancer. The methods of the invention comprise the administration of an effective amount of one or more inhibitors of JNK in combination with the administration of an effective amount of one or more other agents useful for cancer therapy. The invention also provides pharmaceutical compositions comprising one or more inhibitors of JNK in combination with one or more other agents useful for cancer therapy. In particular, the invention is directed to methods of treatment and prevention of cancer by the administration of an effective amount of one or more inhibitors of JNK in combination with standard and experimental chemotherapies, hormonal therapies, bone marrow transplants, stem cell replacement therapies, biological therapies/immunotherapies and/or radiation therapies for treatment or prevention of cancer. Also included are methods of treatment of cancer by the administration of one or more inhibitors of JNK in combination with surgery, alone or in further combination with standard and experimental chemotherapies, hormonal therapies, bone marrow transplants, stem cell replacement therapies, biological therapies/immunotherapies and/or radiation therapies.

JNK inhibitors for use in combination therapy for treating or managing proliferative disorders and cancers

The instant invention describes a method for detecting protein-protein interactions in living organisms and/or cells, said method comprising: (a) synthesizing probe protein fragments from a protein which enables fluorescent or luminescent detection by dissecting the gene coding for the fluorescent or luminescent protein into a least two fragments; (2) constructing fusion proteins consisting of the probe protein fragments linked to protein domains that are to be tested for interactions; (3) coexpressing the fusion proteins; and (d) detecting reconstitution of the fluorescence or luminescence signal.

Protein fragment complementation assays in whole animals applications to drug efficacy, ADME, cancer biology, immunology, infectious disease and gene therapy

The present invention provides protein single-color and multi-color protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter such as monomeric enzymes and fluorescent proteins, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. The development of such assays provides for a broad, flexible and biologically relevant platform for drug discovery.

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San Ramon, CA, United States of America

John K Westwick