VALIDATING BIOLOGICAL MOLECULES AS DRUG TARGETS BY METAL-ION CHELATES IN ANIMAL TEST MODELS

摘要

A target validation process for testing or validating the physiological importance and/or the therapeutic or pharmacological potential of biological target molecules, notably proteins such as, e.g., receptors and especially 7TM receptors in test animals expressing the biological target molecule with, notably, a silent, engineered metal-ion site. A test animal suitable for use in the present invention is normally a genetically modified animal. At any given time during the development of the test animal or in adult life, it is then possible to turn the biological target molecule (such as, e.g., the receptor) on or off - depending on the engineered site with a pharmacological tool, i.e. a metal-ion chelate formed between a metal-ion and a metal-ion chelator. The thus developed pharmacologically controlled 'knock-out' methods are useful in the evaluation of biological target molecules such as, e.g., proteins as drug targets as well as in the characterization of the physiological role of orphan receptors. The process comprises i) introduction of a silent metal ion site in the biological target molecule to obtain a silent metal ion engineered biological target molecule, ii) in vitro testing of a test compound for its ability to bind to the introduced silent metal ion site in the silent metal ion engineered biological target molecule, iii) optionally, chemically optimising the test compound and/or the biological target molecule to create secondary interaction(s) with chemical groups in the vicinity of the metal ion site in the silent metal ion engineered biological target molecule, iv) optionally, repeating any of steps ii) and iii) to obtain a suitable binding affinity in the in vitro test, v) optionally, chemically optimising the test compound to improve the pharmacokinetic and/or biopharmaceutical properties of the test compound, vi) preparing a genetically modified test animal containing the silent metal ion site engineered biological target molecule, vii) in vivo testing of the optionally optimised test compound in the genetically modified test animal, and monitoring the biochemical, physiological and/or behaviour parameters of the genetically modified test animal.