Inositol pyrophosphates (PP-IPs) are densely
phosphorylated messenger molecules involved in numerous
biological processes. PP-IPs contain one or two pyrophosphate
group(s) attached to a phosphorylated myo-inositol ring. 5PP-IP5 is
the most abundant PP-IP in human cells. To investigate the function
and regulation by PP-IPs in biological contexts, metabolically stable
analogs have been developed. Here, we report the synthesis of a new
fluorinated phosphoramidite reagent and its application for the
synthesis of a difluoromethylene bisphosphonate analog of 5PP-IP5.
Subsequently, the properties of all currently reported analogs were
benchmarked using a number of biophysical and biochemical
methods, including co-crystallization, ITC, kinase activity assays and
chromatography. Together, the results showcase how small structural
alterations of the analogs can have notable effects on their properties
in a biochemical setting and will guide in the choice of the most
suitable analog(s) for future investigations
