Use of EEF1A as biomarker and a method of screening MetAP2 inhibitors

摘要

The invention relates to a method for screening compounds, which inhibit MetAP2 activity, by providing a cellular system or a sample thereof being capable of expressing MetAP2 and/or EEF1A, incubating at least a portion of the system with compounds to be screened, and detecting MetAP2 inhibition by determining EEF1A with N-terminal methionine residue (MetEEF1A). Another object of the invention concerns a method for monitoring physiological and/or pathological conditions, which are caused, mediated and/or propagated by MetAP2 activity, by administering an effective amount of at least a single compound to a mammal in need of such treatment and determining MetEEF1A in a biological sample withdrawn from the mammal. The invention also relates to the use of EEF1A as biomarker.

主权项

1. A method for identifying a compound that has methionine aminopeptidase 2 (MetAP2)-dependent anti-proliferative activity, anti-angiogenic activity or anti-tumor activity, comprising
(a) incubating a cellular system that expresses MetAP2 and eukaryotic translation elongation factor 1 alpha (EEF1A) with a compound that is exogenous in origin to the cellular system, wherein the system is selected from the group consisting of single cells, cell cultures, tissues, organs and mammals, and (b) directly detecting EEF1A with an N-terminal methionine residue (MetEEF1A) in a sample of the cellular system of step (a) and in a sample of a negative control system by use of isoelectric-focusing electrophoresis (IEF), optionally followed by Edman sequence analysis, or mass spectrometry, whereby a statistically significant increase in the presence of MetEEF1A in the test system as compared to the negative control system indicates that the N-terminal methionine of MetEEF1A is not cleaved in the test system by MetAP2, and the test compound is identified as having MetAP2 inhibitory activity, and (c) selecting the compound identified in (b) for further in vitro analysis.