Method for high-throughput screening of compounds and combinations of compounds for discovery and quantification of actions, particularly unanticipated therapeutic or toxic actions, in biological systems

Abstract

The invention enables high-throughput screening of compounds in living systems to detect unanticipated or unintended biological actions. The invention also allows for screening, detection, and confirmation of new indications for approved drugs. Screening and detection of toxic effects of compounds also can be achieved by using the methods of the invention. The methods comprise administering isotope-labeled substrates to a living system so that the label is incorporated into molecules in a manner that reveals flux rates through metabolic pathways thought to be involved in a disease. Comparisons between living systems exposed to compounds and living systems not so exposed reveals the effects of the compounds on the flux rates through the metabolic pathways. Combinations or mixtures of compounds can be systematically screened to detect unanticipated or unintended biological actions, including synergistic actions, in the same manner.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60 / 525,261 filed Nov. 25, 2003, the disclosure of which is incorporated herein by referenced in its entirety.FIELD OF THE INVENTION [0002] The invention relates to methods for screening compounds, combinations of compounds, or mixtures of compounds (i.e., drugs and drug candidates, including chemical entities, whether new or known, and biological factors, whether new or known) for actions in biological systems. The disclosed methods measure and quantify molecular flux rates through metabolic pathways (synthesis and breakdown or input and removal rates from pools of molecules) in vivo as targets of drug action. The disclosed methods are capable of high-throughput, large-scale, automated applications. The methods are particularly applicable to detecting and establishing unanticipated or unintended actions of drugs or drug candidates during drug discovery, d...

AI Technical Summary

Benefits of technology

[0016] Stable isotope-labeled substrates are incorporated into one or more molecules of one or more metabolic pathways of interest. In this manner, the molecular flux rates can be determined by measuring, over specific time intervals, isotopic content and / or pattern or rate of change of isotopic content and / or pattern in the targeted molecules, for example by using mass spectrometry, allowing for the determination of the molecular flux rates within the one or more metabolic pathways of interest, by use of analytic and calculation methods known in the art.

[0017] Alternatively, the use of radiolabeled substrates is contemplated for use in the present invention wherein the radiolabeled substrates are incorporated into one or more molecules of one or more metabolic pathways of interest. In this manner, the molecular flux rates can be determined by measuring radiation and / or radioactivity of the targeted molecules of interest within the one or more metabolic pathways of interest by using techniques well known in the art such as scintillation counting. The molecular flux rates within the one or more metabolic pathways of interest are then calculated, using methods known in the art.

[0018] In another embodiment of the invention, isolated isotopically perturbed molecules are provided, the isotopically perturbed molecules including one or more isotopes. The isolated isotopically perturbed molecules are products of the labeling methods described herein. The isolated isotopically perturbed molecules are collected by sampling techniques known in the art and are analyzed using appropriate analytical tools. In one embodiment of the invention, the isolated isotopically perturbed molecules are comprised of one or more stable isotopes. In another embodiment, the isolated isotopically perturbed molecules are comprised of one or more radioactive isotopes.

[0019] In yet another embodiment of the invention, one or more kits are provided that comprise isotope-labeled precursors and instructions for using them. The kits may contain stable-isotope labeled precursors or radioactive-labeled isotope precursors or both. Stable-isotope labeled precursors and radioactive-labeled isotope precursors may be provided in one kit or they may be separated and provided in two or more kits. The kits may further comprise one or more tools for administering the isotope-labeled precursors. The kits also may comprise one or more tools for collecting samples from a subject.

[0020] In yet another embodiment of the invention, one or more information storage devices are provided that comprise data generated from the methods of the present invention. The data may be analyzed, partially analyzed, or unanalyzed. The data may be imprinted onto paper, plastic, magnetic, optical, or other medium for storage and display.

[0021] In yet another embodiment of the invention, one or more compounds, combinations of compounds, or mixtures of compounds identified and at least partially characterized by the methods of the present invention are contemplated.

Problems solved by technology

The metabolic action of the compound (or combination of compounds or mixture of compounds) on the exposed cell, tissue or organism may be unexpected or unanticipated (based on prevailing biochemical knowledge and concepts about the compound and molecular flux within the pathway) or may be anticipated or expected.

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