Toward a Biorelevant Structure of Protein Kinase C Bound Modulators: Design, Synthesis, and Evaluation of Labeled Bryostatin Analogues for Analysis with Rotational Echo Double Resonance NMR Spectroscopy

摘要: Protein kinase C (PKC) modulators are currently of great importance in preclinical and clinical studies directed at cancer, immunotherapy, HIV eradication, Alzheimer’s disease. However, the bound conformation PKC a membrane environment is not known. Rotational echo double resonance (REDOR) NMR spectroscopy could uniquely address this challenge. REDOR requires strategically labeled, high affinity ligands to determine interlabel distances from which ligand PKC–ligand complex be identified. Here we report first computer-guided design syntheses three bryostatin analogues labeled for analysis. Extensive computer analyses energetically accessible analogue conformations suggested preferred labeling sites identification PKC-bound conformers. Significantly, were synthesized, and, as required analysis, all proved highly potent with affinities (∼1 nM)...