CAJ | 학술논문

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33.04μmol· L-1，ALMWH延长CT 1倍的浓度为223.56μmol· 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3168.4μmol· L-1和152.6μmol· L-1，半数抑制MDA-MB-231增殖的IC50分别为12299.6μmol· L-1和22.2μmol· 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。
목적：대간소진행화학수식개조，병검측화학수식물적항응혈활성화항종류활성。방법용고전산납양화、붕경화납환원법강해보통간소（UFH），병용대위조이배기기탄이아알（DCC）화이갑안기필정（DMAP）위최화제대소획저분자량간소（LMWH）진행을선화수식，획득저항응활성적을선화저분자간소（ALMWH）。분별재소서화MDA-MB-231화MCF-7인유선암세포주검측ALMWH적항응혈활성화항종류세포증식화침습활성。결과ALMWH적항응혈활성현저강저，시수저분자간소（LMWH*）연장응혈시간（CT）1배적농도위33.04μmol· L-1，ALMWH연장CT 1배적농도위223.56μmol· L-1，ALMWH장소서응혈시간연장1배소수적약물농도제고도LMWH*적6배지다。0.1、0.3、0.9、2.7、8.1 mmol· L-1계렬농도적ALMWH화LMWH대인유선암세포MDA-MB-231화MCF-7적증식균표현출현저적제량의뢰성억제작용。단량약적억제인유선암세포증식강도구유명현적차이。LMWH화ALMWH적반수억제MCF-7증식농도（IC50）분별위3168.4μmol· L-1화152.6μmol· L-1，반수억제MDA-MB-231증식적IC50분별위12299.6μmol· L-1화22.2μmol· L-1。화학수식장LMWH억제MCF-7적IC50화억제MDA-MB-231적IC50분별강저료20화560배。ALMWH화LMWH대우MDA-MB-231세포적침습능력구유명현적억제작용，통계학분석미발현이자억제강도상적명현차이。결론결구적화학수식가강저LMWH적항응혈활성，증강기항암세포증식화침습활성。
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.