Highly Potent Bisphosphonate Ligands for Phosphoglycerate Kinase and Protein Binding Studies

Abstract

Several novel α,ω-bisphosphonate analogs (1) of 1,3-bisphospho-D-glyceric acid, 1,3-BPG (2) have been made and their binding to phosphoglycerate kinase, PGK (EC 2.7.2.3) evaluated. Non-scissile methanephosphonic acids replace the two phosphate monoesters of 1,3-BPG. Fluorine substitution in the α-methylene groups of the phosphonic acid analogs (1) markedly improves their binding to PGK as determined by NMR analysis. The best ligands bind some 50-100 times more strongly than does 3-phospho-D-glyceric acid and show a requirement for pK _a3 to be below 6.0 while the presence of a β-carbonyl group (1, Y = CO) seems to be of secondary importance. One analog, 1,1,5,5-tetrafluoropentanebisphosphonic acid, has been linked to adenosine 5′-phosphate and 5′-diphosphate to generate bisubstrate analogs (3) whose affinity for PGK and an X-ray crystal structure of a binary complex with PGK will be discussed.

Keywords::

• Phosphonates
• PGK
• binding studies
• bisubstrate
• pKa difluoromethylenephosphonates