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1.发明公开
公开(公告)号：EP1616186A2
公开(公告)日：2006-01-18
申请号：EP04759337.1
申请日：2004-04-09
申请人： Odyssey Thera Inc.
CPC分类号： C07K14/43595 , C12N15/1055 , G01N33/542 , G01N33/58 , G01N33/582 , G01N33/68 , G01N33/6845 , G01N2333/43595
摘要： The present invention is directed to Protein-fragment Complementation Assays (PCAs) and assay compositions based on fluorescent proteins. The invention provides methods for fragmenting fluorescent proteins and generating mutant fragments with desir4 spectral characteristics for PCA. The invention encompasses assays and compositions based on fluorescent proteins from the species Aequorea, Anemonia and Anthozoa. In particular, the invention is directed to fragments of mutant fluorescent proteins having improved spectral properties over the wild-type proteins. The invention encompasses fragments of mutant versions of A. Victoria green fluorescent protein (GFP), in particular yellow fluorescent proteins (EYFP and super-EYFP), 'Venus', cyan, 'citrine', blue, cyan-green, and photoactivatable variants of GFP. The invention also encompasses red fluorescent PCAs based on Discosoma red fluorescent protein (RFP PCA)and a kindling fluorescent protein PCA (KFP 1 PCA) derived from Anemonia sulcata. Any useful mutation of a fluorescent protein can be engineered into a fragment, generating a wide range of assays useful for drug discovery, target validation, high-throughput screening, high-content screening, pathway mapping, drug mechanism-of-action studies, blosensors, and diagnostics.
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2.发明授权
公开(公告)号：EP1590476B1
公开(公告)日：2012-09-26
申请号：EP04708966.9
申请日：2004-02-06
申请人： Odyssey Thera Inc.
发明人： MICHNICK, Stephen, William, Watson , REMY, Ingrid , MACDONALD, Marnie , LAMERDIN, Jane , YU, Helen , WESTWICK, John, K.
IPC分类号： G01N33/50
CPC分类号： G01N33/5011 , G01N33/5008 , G01N33/5041 , G01N33/542 , G01N33/6845 , G01N2500/02 , Y02A90/26
摘要： 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.
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公开(公告)号：EP1737972B1
公开(公告)日：2012-02-01
申请号：EP05735396.3
申请日：2005-04-11
申请人： Odyssey Thera, Inc.
CPC分类号： C40B30/04 , G01N33/5008 , G01N33/5014 , G01N33/502 , G01N33/5041 , G01N33/68 , G01N33/6845 , G06F19/12 , Y02A90/26
摘要： The present invention provides methods for performing pharmacological profiling of a chemical compound, in particular to improve drug safety and efficacy at an early stage in the drug development process. The chemical compound may be a test compound, drug lead, known drug or toxicant. The compound is profiled against a panel of assays. Preferred embodiments of the invention include high-content assays for protein-protein interactions. The compositions and methods of the invention can be used to identify pathways underlying drug efficacy, safety, and toxicity; and to effect attrition of novel compounds with undesirable or toxic properties. The compositions and methods of the invention can also be used to identify new uses of therapeutic agents, to screen libraries of chemical compounds, to perform lead optimization, and to perform studies of structure-activity relationships in the context of intact cells. The compositions and methods of the invention can be applied to any test compound, drug, drug target, pathway, and therapeutic indication.
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4.发明授权PROTEIN FRAGMENT COMPLEMENTATION ASSAYS TO DETECT BIOMOLECULAR INTERACTIONS 失效EIWEISBRUCHSTÜCKERGÄNZUNGSTESTS用于检测BIOMOLEKULARINTERAKTIONEN
公开(公告)号：EP0966685B1
公开(公告)日：2005-08-10
申请号：EP98901905.4
申请日：1998-02-02
申请人： Odyssey Thera, Inc.
IPC分类号： G01N33/68 , G01N33/58 , G01N33/573
CPC分类号： G01N33/6845 , A01K67/0271 , A01K2267/0331 , A01K2267/0393 , C07K14/43595 , C07K2319/00 , C12N15/1055 , G01N33/5008 , G01N33/542 , G01N33/573 , G01N33/58 , G01N33/581 , G01N33/68 , G01N33/6803 , Y02A90/26
摘要： Compositions for protein fragment complementation assays (PCAs) to detect biomolecular interactions are described and their broad applications illustrated with a large number of enzymes, in particular murine dihydrofolate reductase (DHFR). Fusion peptides consisting of N - and C -terminal fragments of murine DHFR fused to GCN4 leucine zipper sequences were coexpressed in E. coli with endogenous DHFR activity inhibited by trimethoprim. Coexpression of the complementary fusion products restored colony formation. Survival only occurred when both DHFR fragments were present and contained leucine-zipper forming sequences, demonstrating that reconstitution of enzyme activity requires assistance of leucine-zipper formation. DHFR fragment-interface mutants of increasing severity resulted in a sequential increase in E-coli doubling times illustrating successful DHFR fragment reassembly rather than non-specific interactions between fragments. The selection and design criteria are developed for numerous examples of clonal selection, colorimetric, fluorometric and other assays based on enzymes whose products can be measured.
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公开(公告)号：EP1027608B1
公开(公告)日：2010-01-20
申请号：EP99936199.1
申请日：1999-07-30
申请人： Odyssey Thera, Inc.
IPC分类号： G01N33/58 , G01N33/566 , G01N33/68
CPC分类号： G01N33/566 , C12N15/1055 , G01N33/581 , G01N33/68 , G01N2500/00
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公开(公告)号：EP1737972A2
公开(公告)日：2007-01-03
申请号：EP05735396.3
申请日：2005-04-11
申请人： Odyssey Thera, Inc.
IPC分类号： C12Q1/00
CPC分类号： C40B30/04 , G01N33/5008 , G01N33/5014 , G01N33/502 , G01N33/5041 , G01N33/68 , G01N33/6845 , G06F19/12 , Y02A90/26
摘要： The present invention provides methods for performing pharmacological profiling of a chemical compound, in particular to improve drug safety and efficacy at an early stage in the drug development process. The chemical compound may be a test compound, drug lead, known drug or toxicant. The compound is profiled against a panel of assays. Preferred embodiments of the invention include high-content assays for protein-protein interactions. The compositions and methods of the invention can be used to identify pathways underlying drug efficacy, safety, and toxicity; and to effect attrition of novel compounds with undesirable or toxic properties. The compositions and methods of the invention can also be used to identify new uses of therapeutic agents, to screen libraries of chemical compounds, to perform lead optimization, and to perform studies of structure-activity relationships in the context of intact cells. The compositions and methods of the invention can be applied to any test compound, drug, drug target, pathway, and therapeutic indication.
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7.发明授权PROTEIN FRAGMENT COMPLEMENTATION ASSAYS TO DETECT BIOMOLECULAR INTERACTIONS 失效EIWEISBRUCHSTÜCKERGÄNZUNGSTESTS用于检测BIOMOLEKULARINTERAKTIONEN
公开(公告)号：EP0966685B9
公开(公告)日：2006-03-15
申请号：EP98901905.4
申请日：1998-02-02
申请人： Odyssey Thera, Inc.
IPC分类号： G01N33/68 , G01N33/58 , G01N33/573
CPC分类号： G01N33/6845 , A01K67/0271 , A01K2267/0331 , A01K2267/0393 , C07K14/43595 , C07K2319/00 , C12N15/1055 , G01N33/5008 , G01N33/542 , G01N33/573 , G01N33/58 , G01N33/581 , G01N33/68 , G01N33/6803 , Y02A90/26
摘要： Compositions for protein fragment complementation assays (PCAs) to detect biomolecular interactions are described and their broad applications illustrated with a large number of enzymes, in particular murine dihydrofolate reductase (DHFR). Fusion peptides consisting of N - and C -terminal fragments of murine DHFR fused to GCN4 leucine zipper sequences were coexpressed in E. coli with endogenous DHFR activity inhibited by trimethoprim. Coexpression of the complementary fusion products restored colony formation. Survival only occurred when both DHFR fragments were present and contained leucine-zipper forming sequences, demonstrating that reconstitution of enzyme activity requires assistance of leucine-zipper formation. DHFR fragment-interface mutants of increasing severity resulted in a sequential increase in E-coli doubling times illustrating successful DHFR fragment reassembly rather than non-specific interactions between fragments. The selection and design criteria are developed for numerous examples of clonal selection, colorimetric, fluorometric and other assays based on enzymes whose products can be measured.
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8.发明公开
公开(公告)号：EP1590476A2
公开(公告)日：2005-11-02
申请号：EP04708966.9
申请日：2004-02-06
申请人： Odyssey Thera Inc.
发明人： MICHNICK, Stephen, William, Watson , REMY, Ingrid , MACDONALD, Marnie , LAMERDIN, Jane , YU, Helen , WESTWICK, John, K.
CPC分类号： G01N33/5011 , G01N33/5008 , G01N33/5041 , G01N33/542 , G01N33/6845 , G01N2500/02 , Y02A90/26
摘要： 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.
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