Preparation of Phosphatidyl-panthenol by phospholipase D-mediated transphosphatidylation and its anti-inflammatory activity on macrophage-like RAW264.7 cells

Panthenol is known to be pro-vitamin B-5 and show several functions such as wound-healing, anti-oxidative, and anti-inflammatory activities. In this study, panthenol was phosphatidylated via phospholipase D (PLD)-mediated transphosphatidylation and its anti-inflammatory effects were investigated on macrophage-like RAW264.7 cells. By PLD-mediated transphosphatidylation from phosphatidylcholine (PC), hydroxy group of p1 position in panthenol molecule was alternatively formed phosphate ester bond even though panthenol has three of hydroxy groups in the molecule. The yield was 96 mol% under optimum reaction conditions: 50 umol of PC, 1.6 U of PLD, 500 umol of panthenol, 1.6 mL/1.6 mL of ethyl acetate/0.2 M acetate buffer (pH 5.6) ratio, 10 mM of CaCl2 in the buffer, 37 degrees C for 24 h. Phosphatidyl-panthenol (P-panthenol) suppressed mRNA expression level of pro-inflammatory mediators such as IL-6, IL-1 beta, TNF-alpha and COX-2 more than precursor compounds, PC and panthenol, in activated RAW264.7 cells. P-panthenol also suppressed protein secretion levels of IL-6 and IL-1 beta in the culture medium of RAW264.7 cells. These results demonstrate that P-panthenol, which is easily synthesized in high yield, is potential compound for novel anti-inflammatory phospholipid.