CHEMICALLY MODIFIED HEPARINS AS INHIBITORS OF HEPARAN-SULFATE SPECIFIC ENDO-BETA-GLUCURONIDASE (HEPARANASE) OF METASTATIC MELANOMA-CELLS
CHEMICALLY MODIFIED HEPARINS AS INHIBITORS OF HEPARAN-SULFATE SPECIFIC ENDO-BETA-GLUCURONIDASE (HEPARANASE) OF METASTATIC MELANOMA-CELLS
复制标题
DOI：
10.1021/bi00366a050
复制
发表时间：
1986-09-09
期刊：
影响因子：
2.9
通讯作者：
NICOLSON, GL
中科院分区：
文献类型：
--
作者：
IRIMURA, T;NAKAJIMA, M;NICOLSON, GL
目的：探讨硫酸肝素（HS）降解酶- β的意义。-葡糖苷酸酶（肝素酶）在肿瘤侵袭和转移中的作用，为了开发可能的抗转移药物，我们合成了该酶的特异性抑制剂。我们先前发现肝素酶活性与小鼠B16黑色素瘤细胞的肺部定植能力相关，并被肝素抑制[Nakajima， M., Irimura， T., Di Ferante， N., and Nicolson， G.L. (1984) J. Biol.]。化学通报，1999,22(2):1 - 3。在本研究中，对肝素进行化学修饰，以确定其结构的哪些部分负责肝素酶抑制活性，并获得具有最小额外生物效应的肝素酶抑制剂，如抗凝。分别去除肝素中的n -硫酸盐基团和o -硫酸盐基团，得到的游离氨基被乙酰化或生成。肝素在醛酸的羧基上也被还原。采用高速凝胶渗透色谱法和琼脂糖球固定化放射性HS法检测了这些肝素衍生物的肝素酶抑制活性。结果表明，尽管氨基葡萄糖残基上的n -硫酸盐和o -硫酸盐基团以及脲酸残基上的羧基对肝素酶的抑制作用很重要，但它们并不是完全抑制肝素酶活性所必需的。将高度转移的B16-BL6黑色素瘤细胞与N-乙酰化N-去硫肝素、N-生成的N-和o -去硫肝素或羧基还原肝素一起培养，并静脉注射给同体C57BL/6小鼠，实验中黑色素瘤肺转移的数量显著减少。
To determine the significance of the heparan sulfate (HS) degradative endo-.beta.-glucuronidase (heparanase) in tumor invasion and metastasis and to develop possible antimestatic agents, we synthesized specific inhibitors of this enzyme. We previously found that heparanase activity correlates with the lung colonization abilities of murine B16 melanoma cells and is inhibited by heparin [Nakajima, M., Irimura, T., Di Ferante, N., and Nicolson, G.L. (1984) J. Biol. Chem. 259, 2283-2290]. In this study, heparin as chemically modified in order to determine which portions of its structure are responsible for heparanase inhibitory activity and to obtain heparanase inhibitors that have minimal additional biological effects, such as anticoagulation. N-Sulfate groups and O-sulfate in heparin were removed separately, and the resultant free amino groups were acetylated or resulfated. Heparin was also reduced at the carboxy groups of uronic acid. The heparanase inhibitory activities of these heparin derivatives were examined by high-speed gel-permeation chromatography and by the use of radioactive HS immobilized on agarose beads. The results indicated that although N-sulfate and O-sulfate groups on glucosamine residues, and carboxy groups on uronimc acid residues, are important for heparanase inhibition, they are not essential for full activity. When highly metastatic B16-BL6 melanoma cells were incubated with N-acetylated N-desulfated heparin, N-resulfated N- and O-desulfated heparin, or carboxyl-reduced heparin and injected intravenously to syngenic C57BL/6 mice, significant reductions in the numbers of experimental melanoma lung metastases occurred.