Investigating the Solubility and Activity of a Novel Class of Ice Recrystallization Inhibitors

Abstract

O-aryl-β-D-glucosides and N-alkyl-D-gluconamides are two classes of effective ice recrystallization inhibitors (IRIs), however their solubilities limit their use in cryopreservation applications. Herein, we have synthesized and assessed phosphonate analogues of small-molecule IRIs as a method to improve their chemical and physical properties. Four sodium phosphonate compounds 4–7 were synthesized and exhibited high solubilities greater than 200 mM. Their IRI activity was evaluated using the splat cooling assay and only the sodium phosphonate derivatives of α-methyl-D-glucoside (5-Na) and N-octyl-D-gluconamide (7-Na) exhibited an IC50 value less than 30 mM. It was found that the addition of a polar sodium phosphonate group to the alkyl gluconamide (1) and aryl glucoside (2) structure decreased its IRI activity, indicating the importance of a delicate hydrophobic/hydrophilic balance within these compounds. The evaluation of various cation-phosphonate pairs was studied and revealed the IRI activity of ammonium and its ability to modulate the IRI activity of its paired anion. A preliminary cytotoxicity study was also performed in a HepG2 cell line and phosphonate analogues were found to have relatively low cytotoxicity. As such, we present phosphonate small-molecule carbohydrates as a biocompatible novel class of IRIs with high solubilities and moderate-to-high IRI activities.

Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield

“…ice growth) modes of damage. This includes ice binding proteins, 10 controlled nucleation, 11,12 ice recrystallisation inhibitors, 13–16 and macromolecular cryoprotectants. 17–19 Despite the success of all these additives, they do not target the biochemical damage induced by hypothermic temperatures, nor the damage mechanisms which remain and impair post-thaw function.…”