Structure-activity studies on inhibition of choline uptake by a mouse brain synaptosomal preparation: basic data.

Over 80 substances were studied for their inhibition of high-affinity uptake of [3H]choline into a mouse brain synaptosomal fraction. Kinetic experiments tested a number of them for competitive behavior. A minimal provisional model for the choline uptake process is envisioned that is consistent with current data and with relevant observations in the literature. There are two hydrophilic anionic sites on the choline transporter that are separated from each other by a cationic hydrophobic domain. Association of choline in a Na+-dependent manner with one or both of the sites is necessary for the transport of choline to take place. The choline binding anionic sites are sufficiently large and/or flexible to allow attachment of cationic moieties larger than choline. The cationic hydrophobic domain of the transporter is flexible, probably tending to planarity. The length of the hydrophobic region between the anionic sites is approximately that of 10 extended methylene groups, and the minimal width is approximated by the distance across the condensed ring system of chlorpromazine. The probability of attachment of the highly hydrophilic choline to its binding sites is increased both by hydrogen-bonding to a proton-acceptor within the anionic sites and by repulsion from the cationic hydrophobic region. A number of substances that potently and competitively inhibit high affinity choline uptake possess quaternary ammonium groups and neutral or negatively charged lipophilic groups. In general, substances in which two quaternary ammonium groups are separated by an appropriately configured hydrophobic group and which can combine with both anionic sites and the hydrophobic region between them are more potent inhibitors than monoquaternary substance with the same or similar groups. However, substances with a single high-affinity quaternary group and an appropriately structured hydrophobic group, e.g. the trimethoxy-3-butynyl quaternary ammonium compounds, possess inhibitory efficacies similar to those shown by the most potent bisquaternaries. The above suggests that further delineation of the characteristics of the structures of the above sites of the transporter could lead to devisal of more potent reversible inhibitors of choline uptake than now are available.