Concepts for improved regioselective placement of O-sulfo, N-sulfo, N-acetyl, and N-carboxymethyl groups in chitosan derivatives☆

Abstract In the present paper a new strategy has been studied to introduce solely or in combination N -sulfo, O -sulfo, N -acetyl, and N -carboxymethyl groups into chitosan with highest possible regioselectivity and completeness and defined distribution along the polymer chain. The aim was to generate compounds having lowest toxicity for determining the pharmacological structure–function relationships among different backbone structures and differently arranged functional groups compared to those of heparin and heparan sulfate. The water-soluble starting material, chitosan, with a degree of acetylation (DA) of 0.14 and a molecular weight of 29 kD, allows one to apply most of the known reactions of chitosan as well as some reactions of heparin chemistry successfully and with improved regioselectivity and completeness. On the other hand, a number of these reactions were not successful by application to water-soluble high-molecular-weight chitosan (DA 0.45 and 150 kD). The starting material showed statistical N -acetyl ( N -Ac) distribution along the polymer chain according to the rules of Bernoulli, with highest abundance of the GlcNAc–GlcNAc diad along with a lower abundance of triads, tetrads, and pentads. The space between the N -Ac groups was filled up in homogeneous reactions by N -sulfo and/or N -carboxymethyl groups, which also resulted in a Bernoulli statistical distribution. The N-substitution reaction showed highest regioselectivity and completeness with up to three combined different functional groups. The regioselectivity of the 3- O -sulfo groups was improved by regioselective 6-desulfation of nearly completely sulfated 3,6-di- O -sulfochitosan. By means of desulfation reactions, all of the possible intermediate sulfated products are possible. 6- O -Sulfo groups can also be introduced with highest regioselectivity and completeness, and a number of partially 6-desulfated products are possible.