Cubist Pharmaceuticals, Incorporated

Other

Jiansu Zhang

Jianshi Tao

Xiaoyu Shen

Francis P Tally

Paul R Schimmel

Ronald P Gladue

Paul L Gallant

John B Cheng

Philip A Wendler

Gene Connelly

Susan A Martinis

Kuldeep S Neote

Baiyong Li

Paul H Bauer

The present invention relates to the discovery that PDGis the nautral ligand for orphan receptor CRTH2, which interaction, it will now immediately be recognized, is important in the development of valuable pharmaceuticals. The present invention is therefore related to methods for screening for therapeutic compounds useful in the treatment of PGD-related disorders such as allergy, asthma and inflammation. Appropriate assay methodology is also disclosed.

Methods for the identification of compounds useful for the treatment of disease states mediated by prostaglandin D2

The invention comprises methods useful within a larger process for identifying compounds and/or designing further compounds with activity to produce a desired phenotype (for example, growth inhibition) in cells whose target cell component is the subject of certain studies to identify such compounds. The invention employs constructed cells comprising a regulable gene encoding a biomolecule which modulates (inhibits or activates) in vivo the function of a target component of the cell which can be an enzyme, for example, methionyl-tRNA synthetase of . The process incorporates methods for identifying biomolecules that bind to a chosen target cell component in vitro, methods for identifying biomolecules that also bind to the chosen target and modulate its function intracellularly, causing a phenotypic effect. The intracellular effect of a biomolecule can be tested in cell culture, or tested after introduction of the constructed cells into a host mammal in vivo, and methods for identifying compounds that compete with the biomolecules for sites on the target in competitive binding assays. Compounds identified by the series of steps in this process are candidates for drugs with the desired activity on the cell. Targets for which such compounds can be identified are validated as being essential to a phenotype of the cell.

Method for identifying validated target and assay combinations for drug development

A system has been constructed which is suitable for tetracycline-inducible gene expression in Gram-positive bacteria such as and . The replicon/host gene expression system is tightly regulated, can be used in complex as well as minimal media, and can produce a high level of gene expression upon induction, with a variety of gene products. The gene expression system is suitable for production of products toxic to the host cells, and can be used, for example, for the analysis of gene expression and gene products in Gram-positive bacteria, and in a test of the effect of a peptide or polypeptide inhibitor of an enzyme on the growth of cells in culture or during infection of an animal.

Regulable gene expression in gram-positive bacteria

Isolated and/or recombinant nucleic acids encoding mycobacterial seryl-tRNA synthetase have been characterized. Recombinant DNA constructs and vectors having a sequence which encodes mycobacterial seryl-tRNA synthetase have been made, and can be used for the construction of tester strains as well as for the production of isolated and/or recombinant seryl-tRNA synthetases. These enzymes or portions thereof are useful in the biochemical separation of serine and quantification of serine or ATP, and for producing antibodies useful in the purification and study of the enzyme, for example. Host cells and methods useful for producing recombinant mycobacterial seryl-tRNA synthetases are described, as are tester strains, which are cells engineered to rely on the function of the tRNA synthetase encoded by an introduced cloned gene. Tester strains can be used to identify inhibitors of the essential tRNA synthetase enzyme encoded by the introduced cloned gene, and thus provide a means to assess the antimicrobial effect and specificity of the inhibitor without employing slow-growing, pathogenic strains of mycobacteria, such as Mycobacterium tuberculosis.

Recombinant mycobacterial seryl-tRNA synthetase genes, tester strains

Growing community of inventors

Roslindale, MA, United States of America

Jiansu Zhang