Heparanase activity

CR Parish, C Freeman, KJ Brown, DJ Francis, WB Cowden. Identification of sulfated oligosaccharide-based inhibitors of tumor growth and metastasis using novel in vitro assays for angiogenesis and heparanase activity. Cancer Res 59 3433-3441, 1999. 

Miao, H. Q., Elkin, M., Aingorn, E., Ishai-Michaeli, R., Stein, C. A., and Vlodavsky, 1. (1999). Inhibition of heparanase activity and tumor metastasis by laminarin sulfate and synthetic phosphorothioate oligodeoxynudeotides. Int. J. Cancer 83, 424-d31. 

Heparanase and type IV collagenase activities were assessed by a method described previously [17]. Briefly, H-labeled heparin sulfate (HS) was incubated at 37°C with partially purified heparanase in the presence or absence of chitin derivatives. [ H]HS degradation products in the supernatant were analyzed by size-exclusion chromatography. Heparanase activity was determined by measuring the radioactivity of each fraction. H-labeled type IV collagen was incubated at 37°C for 48 h in the presence or absence of chitin derivatives. Type IV collagenolytic activity was calculated from the radioactivity in the supernatant. 

Total PA activity Total PA activity Collagenase 1 Collagenase 1 CB-Iike enzyme Trypsin-like enzyme Chymotrypsin-like enzyme 92-kDa gelatinase Heparanase... 

Antiangiogenic activity as a heparanase inhibitor, HCII-mediated anticoagulant Preclinical studies inhibitor of tumor metastasis, tumor growth, and angiogenesis 38, 39... 

Various biochemical methods are available for measuring heparanase and other glycosidase activities. The former can be assayed by a method that makes use of a solid-phase substrate for heparanase, such as chemically-modified heparan sulphate coupled covalently at its reducing terminal saccharide to agarose gel beads. The reader is referred to Nakagima et al. (1986). 

Nakagima, M., Irimura, T. and Nicolson, G. L. (1986). A solid-phase substrate of heparanase its application to assay of human melanoma for heparan sulfate degradative activity. Anal. Biochem. 157, 162-171. 

Oligomerization of a 7-amino-2,6-anhydro-7-deoxy-heptonic acid results in mimetics of (1 6)-linked sugars with a three-atom instead of a two-atom linker. The sulfated tetramer 24 (O Scheme 7) turned out to exhibit /x-molar activity in the protection of MT2 cells from HIV infection [33], and was also shown to provide heparanase inhibitory activity [34]. With different location of the amide bond, a still benzyl protected /3(1 6)-linked octamer of glucosyl-uronic acid-methyl amine (Gum [35]) 25 was synthesized [36]. 

J. K. Fairweather, E. Hammond, and V. Ferro, Synthesis and heparanase inhibitory activity of sulfated mannooligosaccharides related to the antiangiogenic agent PI-88, Bioorg. Med. Chem., 16 (2008) 699-799. 

Figure 9 Influence of sulfated chitin derivatives on heparanase and type IV collagenase activities by B16-BL6 melanoma cells V-pH]acetylated HS (5 xg) was incubated at 37°C for 3 h with 26.7 jLg of partially purified heparanase in 50 p,L of 0.2 M sodium acetate buffer, pH 5.6, in the presence or absence of chitin derivatives. The H-labeled type IV collagen film was treated at 37 C for 3 h with or without chitin derivatives and incubated with B16-BL6 cells (5 X lO ) suspended in 400 p.L of a medium containing 0.5% BSA for 48 h.

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