公开(公告)号：EP1442296A2

公开(公告)日：2004-08-04

申请号：EP02768394.5

申请日：2002-08-02

申请人： Merck & Co., Inc.

发明人： INGLESE, James ,  FERRER, Marc ,  HAMILTON, Aaron, C.

IPC分类号： G01N33/53 ,  G01N33/566 ,  C12Q1/34 ,  C12Q1/42

CPC分类号： G01N33/573 ,  G01N33/542

摘要： The invention is a method for detecting a reaction product which signals the presence of a reaction product inducer such as an enzyme. The method enables the recognition of epitopes that form the basis of a detection strategy without the need for specific antibodies to the epitope. In the method, a directly or indirectly labeled modular domain and a biotinylated form of the cognate peptide ligand are used as the basis for a measurable interaction. The peptide ligand can be masked by modifications through, for example, phosphorylation of the Ser or Thr residue, or extension of the amino acid sequence beyond the C-terminal Val. Because the masked residues are critical to binding of the labeled modular domain, masking of at least one of the residues prevents binding. Upon treatment of the masked residue by the appropriate enzyme, (e.g., treatment of the phosphorylated residue with a phosphatase enzyme, or treatment of the extended residue with a protease enzyme, the peptide is converted to the original unmasked ligand that is capable of binding to the labeled modular domain and forming a measurable complex.

公开(公告)号：EP1578356A2

公开(公告)日：2005-09-28

申请号：EP03743199.6

申请日：2003-02-23

申请人： Merck & Co., Inc.

发明人： ESPESETH, Amy, S. ,  FERRER, Marc ,  FLORES, Osvaldo, A. ,  HAZUDA, Daria, J. ,  INGLESE, James ,  MILLER, Michael, D. ,  REGISTER, Bruce ,  SHI, Xiao-Ping ,  SIMON, Adam, J. ,  ZUCK, Paul, D.

IPC分类号： A61K6/00

CPC分类号： C07K14/47 ,  C07K14/4702 ,  C07K14/4711 ,  C07K2319/00 ,  C07K2319/50 ,  G01N2500/10

摘要： The present invention provides DNA constructs, genetically engineered host cells, and methods for identifying inhibitors of amyloid precursor protein (APP) processing. The methods provide for the convenient identification, in a single assay, of inhibitors of β-secretase and Ϝ-secretase as well as other forms of APP processing. The methods rely on fusion proteins of APP and transcription factors in which APP processing releases the transcription factors, allowing the transcription factors to activate transcription of a reporter gene. Inhibitors are identified as substances that block or diminish transcription factor release from the fusion protein, thereby causing a diminution of reporter gene readout.