Tilorone-induced lysosomal storage of sulphated glycosaminoglycans can be separated from tilorone-induced enhancement of lysosomal enzyme secretion.
This investigation deals with a drug side-effect. The immunomodulatory drug tilorone (2,7-bis[2-(diethylamino)ethoxy]fluoren-9-one) and congeners induce lysosomal storage of sulphated glycosaminoglycans (GAGs) in animals and in cultured cells. At high tilorone concentrations, GAG storage in cultured fibroblasts was previously reported to be accompanied, and presumably caused by, disturbance of intracellular targeting of lysosomal enzyme precursors, which leads to enhanced secretion and thus loss of lysosomal enzymes. The purpose of the present study was to examine whether the GAG storage induced in cultured bovine fibroblasts by low tilorone concentrations is also accompanied by enhanced lysosomal enzyme release. Enhanced secretion of beta-hexosaminidase (EC 3.2.1.52) was taken as indicating the intracellular mistargeting of lysosomal enzyme precursors. Dose-response curves were established for (a) the intracellular accumulation of 35S-GAGs and (b) the release of beta-hexosaminidase after exposure (72 hr) to tilorone (1-35 microM). For positive controls, the classical lysosomotropic agents NH4Cl (1-30 mM) and chloroquine (1-60 microM) were used. With NH4Cl, 35S-GAG storage was accompanied by enhanced enzyme release throughout the concentration range (EC50 at 3.3 mM for either effect). With chloroquine, low concentrations (< or = 5 microM) caused a small increase in 35S-GAG accumulation without abnormal enzyme secretion; at higher concentrations both drug effects were produced (EC50 around 15 microM for either effect). With tilorone, low concentrations (< or = 5 microM) caused marked 35S-GAG accumulation without enhancement of enzyme release. The EC50 for tilorone-induced 35S-GAG storage was 3 microM, as opposed to 15 microM for enzyme release. The results indicate that GAG storage induced by low concentrations of tilorone is due to mechanisms other than mistargeting and loss of lysosomal enzymes. On the basis of previous results it may be hypothesized that tilorone and other symmetrically substituted dicationic compounds form complexes with the polyanionic GAG chains and thereby impair their enzymic degradation.
Lüllmann-Rauch
R
R
Department of Anatomy, University of Kiel, Germany.
Pods
R
R
Von Witzendorff
B
B
D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000648:ultrastructure; D006025:Glycosaminoglycans / Q000378:metabolism*; D008247:Lysosomes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D009083:Mucopolysaccharidoses / Q000378:metabolism; D013994:Tilorone / Q000494:pharmacology*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D006025:Glycosaminoglycans; D000643:Ammonium Chloride; D002738:Chloroquine; D001619:beta-N-Acetylhexosaminidases; D013994:Tilorone