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In the fragment-based drug design approach, methods for rapid identification of the ligand-binding site are essential. The INPHARMA experiment, a ligand-observed NMR experiment based on the nuclear Overhauser effect, is tested here with fragment-like molecules and the glycogen phosphorylase enzyme that contain multiple binding sites. The results illustrate the potential of the method for the FBDD process and demonstrate that the INPHARMA experiment is particularly useful to study the binding specificity of the fragments and to assess the ligand binding mode in the presence of a reference ligand.
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