Preparation of Nicotinamide Riboside

Description

1- ( β-D-Ribofuranosyl ) -nicotinamide [nicotinamide riboside (NR) ] is a nucleoside, which incorporates nicotinamide and ribose into a single structural motif where the positively charged pyridinium moiety is balanced with a negatively charged counter ion (Formula A) . The nature of the counter ion depends on the synthesis or production procedure. The most common counter ions include, but are not limited to, halide (CI-, Br~) , acetate, phosphate and trifluoromethane- sulfonate. 

Nicotinamide riboside (NR)  naturally occurs in yeast, bacteria, and mammals. Therefore, often, nicotinamide riboside is produced biotechnologically from yeast.

Preparation of Nicotinamide Riboside

Preparation of nicotinamide riboside from NAD+ by a two-stage process (ZrCl4-promoted NAD+ primary hydrolysis followed by secondary hydrolysis)

Stage one: In a 250-ml flask, to a mixture of NAD+ (0.3317 gr) and ZrCl4 (0.582 gr) , 100 ml of deionized water was added and the reaction was heated to 80 °C for 30 minutes. Over this time, full conversion of the NAD to β-nicotinamide mononucleotide was observed by HPLC .

Stage two: 1.8 gr of calcium L-ascorbate dihydrate was added to the reaction mixture and heating was continued for further 2 days at 80 °C.

Samples from the reaction mixture were analyzed every 4-5 hours by HPLC and NMR. Formation of the desired NR was detected albeit in low concentration (lower than 3% yield) due to extensive decomposition of NR to ribose and nicotinamide. HR-MS spectrum of the product of Stage 2 is with MW of 255.16. The yield was determined by comparing the abundance of the desired peak to the abundance of the internal standard.