Methods of identifying compounds that target trna splicing endonuclease and uses of said compounds as anti-proliferative agents

Abstract

The present invention relates to a method for screening and identifying compounds that modulate the activity tRNA splicing endonuclease. In particular, the invention provides assays for the identification of compounds that inhibit animalia tRNA splicing endonuclease. The methods of the present invention provide a simple, sensitive assay for high-throughput screening of libraries of compounds to identify pharmaceutical leads useful for treating and/or preventing cancer.

Description

1. INTRODUCTION [0001] The present invention relates to a method for screening and identifying compounds that modulate the activity of an animialia tRNA splicing endonuclease. In particular, the invention provides assays for the identification of compounds that inhibit or reduce the activity of an animalia tRNA splicing endonuclease. The methods of the present invention provide a simple, sensitive assay for high-throughput screening of libraries of compounds to identify pharmaceutical leads useful for preventing, treating, managing or ameliorating cancer or one or more symptoms thereof. 2. BACKGROUND OF THE INVENTION [0002] 2.1 Cancer and Neoplastic Disease [0003] Cancer is the second leading cause of death in the United States. The American Cancer Society estimated that in 2001, there would be 1.3 million new cases of cancer and that cancer will cause 550,000 deaths. Overall rates have declined by 1% per year during the 1990s. There are 9 million Americans alive who have ever had c...

AI Technical Summary

Benefits of technology

[0027] The effect of a compound on the activity of an animalia tRNA splicing endonuclease may be determined utilizing a fluorescence polarization-based assay. In such an assay, a fluorescently labeled substrate for an animalia tRNA splicing endonuclease is contacted with an animalia cell-free extract or a purified animalia tRNA splicing endonuclease and a compound or member of a library of compounds; and the fluorescent polarized light emitted is measured utilizing techniques well-known to one of skill in the art or described herein, wherein an alteration in the fluorescently polarized light emitted relative to a control or the absence of the compound or the member of a library of compounds indicates that the compound or member of a library of compounds modulates animalia tRNA splicing endonuclease activity.

[0028] Further, the effect of a compound on the activity of an animalia tRNA splicing endonuclease may be determined utilizing a tRNA endonuclease suppression assay. In such an assay, a host cell is engineered to contain a reporter gene and a suppressor tRNA, wherein the reporter gene construct comprises a reporter gene with a nonsense codon in its open reading frame such that the open reading frame is interrupted

Problems solved by technology

All of these approaches pose significant drawbacks for the patient.

Surgery, for example, can be contraindicated due to the health of the patient or can be unacceptable to the patient.

Additionally, surgery might not completely remove the neoplastic tissue.

Radiation therapy is effective only when the irradiated neoplastic tissue exhibits a higher sensitivity to radiation than normal tissue, and radiation therapy can also often elicit seriou

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