A systematic analysis of DMTMM vs EDC/NHS for ligation of amines to hyaluronan in water

Abstract

The activation of carboxyl groups with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS) for amide formation is the standard method for amine ligation to hyaluronan (HA), and a very well established wide-ranging bioconjugation method. In this paper we compare 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) to EDC/NHS activation chemistry for HA ligation using an array of substrates including small, large and functional molecules. For all the substrates tested DMTMM yields were superior at parity of feed ratio. DMTMM chemistry resulted effective also in absence of pH control, which is essential for EDC/NHS conjugation. Overall our results demonstrate that DMTMM is more efficient than EDC/NHS for ligation of amines to HA and does not require accurate pH control or pH shift during the reaction to be effective. DMTMM-mediated ligation is a new promising chemical tool to synthesize HA derivatives for biomedical and pharmaceutical applications.

Keywords: Amide formation; Bioconjugates; DMTMM; EDC/NHS; Hyaluronan modification.