DRUG-INDUCED LYSOSOMAL STORAGE OF SULFATED GLYCOSAMINOGLYCANS - STUDIES ON THE UNDERLYING STRUCTURE-ACTIVITY-RELATIONSHIPS

Some immunomodulatory drugs have previously been shown to induce lysosomal storage of sulfated glycosaminoglycans (sGAG) in intact organisms and cultured cells. These compounds consist of a planary aromatic ring system and two symmetric side chains each carrying a protonizable nitrogen. The purpose of this study was to test a larger collection of such compounds for their potencies to induce lysosomal storage of sGAG in cultured fibroblasts of rat cornea. The cells were exposed (72 h) to various compounds differing with respect to the aromatic ring system or the side chains. Lysosomal sGAG-storage was demonstrated by selective cytochemical staining with cuprolinic blue. The threshold concentration, i.e., the concentration necessary to induce cuprolinic blue-positive cytoplasmic inclusions in at least 1% of the cells, was determined for each compound. The threshold concentrations were distributed over a range of 0.3-30 mu M. It Should be emphasized that the threshold concentration of a given compound is not a constant, but depends on the volume of cell culture medium per surface area of cell monolayer, since the lysosomal accumulation towers the initial drug concentration in the medium. If the ratio of medium volume : cell monolayer surface is increased as compared with standard cell culture conditions, the threshold concentration will be lowered. The compounds were ranked according to their threshold concentrations as determined under standard conditions. The following conclusions can be drawn from the ranking: the type of the central aromatic ring system and the distance between the ring system and the protonizable nitrogen atoms of the side chains influence the potency to induce lysomomal sGAG-storage. Regarding the ring system, the potency decreases as follows: acridine similar to anthrachinone > fenfluorenone similar to fenfluorene > xanthenone; xanthene > dibenzofuran similar to dibenzothiophene. In intact organisms, these structure-activity relationships may be superimposed by drug metabolism and pharmacokinetic factors.