南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (01): 40-.

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化学修饰赋予肝素低抗凝血活性和高抗肿瘤活性

梁颖，李立标，张配，吴成柱，赵素容，张倩雯，刘浩，蒋志文   

  • 出版日期:2015-01-20 发布日期:2015-01-20

Chemical modification endows heparin with low anticoagulant and high antineoplastic
activities

  • Online:2015-01-20 Published:2015-01-20

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摘要： 目的对肝素进行化学修饰改造，并检测化学修饰物的抗凝血活性和抗肿瘤活性。方法用高碘酸钠氧化、硼氢化钠还原法
降解普通肝素（UFH），并用大位阻二环己基碳二亚胺（DCC）和二甲氨基吡啶（DMAP）为催化剂对所获低分子量肝素（LMWH）
进行乙酰化修饰，获得低抗凝活性的乙酰化低分子肝素（ALMWH）。分别在小鼠和MDA-MB-231和MCF-7人乳腺癌细胞株检
测ALMWH的抗凝血活性和抗肿瘤细胞增殖和侵袭活性。结果ALMWH的抗凝血活性显著降低，市售低分子肝素（LMWH*）
延长凝血时间（CT）1倍的浓度为33.04 μmol·L-1，ALMWH延长CT 1倍的浓度为223.56 μmol·L-1，ALMWH 将小鼠凝血时间延
长1倍所需的药物浓度提高到LMWH*的6 倍之多。0.1、0.3、0.9、2.7、8.1 mmol·L-1系列浓度的ALMWH和LMWH对人乳腺癌
细胞MDA-MB-231和MCF-7的增殖均表现出显著的剂量依赖性抑制作用。但两药的抑制人乳腺癌细胞增殖强度具有明显的
差异。LMWH和ALMWH的半数抑制MCF-7 增殖浓度（IC50）分别为3168.4 μmol·L-1 和152.6 μmol·L-1，半数抑制MDAMB-
231增殖的IC50分别为12299.6 μmol·L-1和22.2 μmol·L-1。化学修饰将LMWH抑制MCF-7的IC50和抑制MDA-MB-231的
IC50分别降低了20和560倍。ALMWH和LMWH对于MDA-MB-231细胞的侵袭能力具有明显的抑制作用，统计学分析未发现
二者抑制强度上的明显差异。结论结构的化学修饰可降低LMWH的抗凝血活性，增强其抗癌细胞增殖和侵袭活性。

Abstract: Objective To evaluate the anticoagulant and antineoplastic activities of chemically modified low-molecular-weight
heparin (LMWH). Methods LMWH obtained by splitting unfractionated heparin (UFH) with sodium periodate oxidation and
sodium borohydride reduction was subjected to acetylation catalyzed by DCC and DMAP to produce acetylated LMWH
(ALMWH). The anticoagulant activity of ALMWH was determined in mice, and its antiproliferative and anti-invasion
activities was assessed in human breast cancer cells MDA-MB-231 and MFC-7. Results The anticoagulant activity of LMWH
was decreased significantly after acetylation. The concentrations of commercial LMWH* and ALMWH for doubling the
coagulation time (CT) were 33.04 μmol/L and 223.56 μmol/L, respectively, and the IC50 of ALMWH for doubling CT was 6
times of that of LMWH*. ALMWH and LMWH at 0.1, 0.3, 0.9, 2.7 and 8.1 mmol/L both significantly inhibited the proliferation
of MDA-MB-231 and MCF-7 cells in a concentration-dependent manner, but ALMWH produced stronger inhibitory effects.
The IC50 of LMWH and ALMWH for inhibiting cell proliferation was 3168.4 μmol/L and 152.6 μmol/L in MCF-7 cells, and
12299.6 μmol/L and 22.2 μmol/L in MDA-MB-231 cells, respectively. ALMWH and LMWH all markedly suppressed the
invasion of MDA-MB-231 cells with comparable effects. Conclusion Chemical modification of structure can endow LMWH
with a low anticoagulant and high antiproliferative activities.