Inhibition of amyloid A amyloidogenesis in vivo and in tissue culture by 4-deoxy analogues of peracetylated 2-acetamido-2-deoxy-alpha- and beta-d-glucose: implications for the treatment of various amyloidoses.
Two novel sugars, 2-acetamido-1,3,6-tri-O-acetyl-2,4-dideoxy-alpha- and beta-D-xylo-hexopyranoses, have been synthesized and their effects on heparan sulfate biosynthesis using primary mouse hepatocytes in tissue culture have been assessed. At concentrations of 0.1 and 1.0 mmol/L a mixture of both anomers significantly inhibited the biosynthesis of heparan sulfate by 60% and 99%, respectively. At 1.0 mmol/L the average molecular weight of the heparan sulfate synthesized is reduced from 77 kd to 40 kd. The biosynthetic inhibition is apparent within 1 hour (the earliest time point examined) of exposure of the hepatocytes to the analogues and appears virtually complete throughout a 24-hour incubation period. Using a radiolabeled version of the beta-anomer we demonstrate that the analogue is incorporated into growing heparan sulfate chains. The nature of the analogue, the quantity of analogue isotope incorporated, and the reduction in the size of the heparan sulfate polysaccharide are consistent with UDP activation and incorporation of the analogue and truncation of the growing heparan sulfate chain. At 0.1 mmol/L, and in the presence of a constant concentration of serum amyloid A (the precursor to AA amyloid), each analogue inhibited amyloid deposition (by 95 to 99%) in a tissue culture model of AA amyloidogenesis. At 6 mg/dose twice daily each analogue inhibited in vivo splenic AA amyloid deposition by 65 to 70% when using a rapid induction model of mouse AA amyloidogenesis. These data indicate that polysaccharides, such as heparan sulfate, play an integral part in the pathogenesis of AA amyloid deposition, and potentially other forms of amyloid. These data support our previous work that demonstrated that agents that mimic aspects of heparan sulfate structure and that interfere with heparan sulfate:amyloid protein binding inhibit AA amyloid deposition. They emphasize that heparan sulfate likely plays a critical role in amyloidogenesis, and compounds that interfere with heparan sulfate biosynthesis may provide leads for the development of anti-amyloid therapeutic agents.
Kisilevsky
Robert
R
Department of Pathology, Queen's University and The Syl and Molly Apps Research Center, Kingston General Hospital, Kingston, Ontario, Canada. kisilevsky@cliff.queensu.ca
Szarek
Walter A
WA
Ancsin
John B
JB
Elimova
Elena
E
Marone
Sandra
S
Bhat
Shridhar
S
Berkin
Ali
A
D000107:Acetylation; D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology* / Q000627:therapeutic use; D000686:Amyloidosis / Q000188:drug therapy*; D000818:Animals; D002236:Carbohydrate Conformation; D002460:Cell Line; D005260:Female; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D008099:Liver / Q000187:drug effects; D008264:Macrophages; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D015394:Molecular Structure; D000685:Serum Amyloid A Protein / Q000037:antagonists & inhibitors / Q000502:physiology*; D013154:Spleen / Q000187:drug effects
D000685:Serum Amyloid A Protein; D005944:Glucosamine; D000117:Acetylglucosamine