南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (4): 613-620.doi: 10.12122/j.issn.1673-4254.2021.04.20

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阿霉素-甘草酸分子复合物的制备及体外抗肿瘤活性

吕雪丽，刘 媛，祝瑶露，赵博欣，魏 理，李国锋   

  • 出版日期:2021-04-20 发布日期:2021-04-30

Construction of an adriamycin-glycyrrhizin molecular complex and assessment of its anti-tumor activity

  • Online:2021-04-20 Published:2021-04-30

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摘要： 目的 以中药组份甘草酸为载体，按照阿霉素与甘草酸的不同比例制备阿霉素-甘草酸分子复合物（ADR-GL复合物），探究其在pH=7.4下的溶解度差异并初步评价其抗肿瘤效果。方法 按照阿霉素与甘草酸摩尔比2∶1，1∶1，1∶2的量（ADR∶GL=2∶1，1∶1，1∶2）投料，采用旋蒸法制备ADR-GL分子复合物并经红外光谱（FT-IR）和差式扫描量热（DSC）分析鉴定所得固体复合物。将ADR-GL复合物溶解于pH=7.4的磷酸盐缓冲液中，过夜超声后3000 r/min离心5 min得ADR-GL复合物饱和溶液，采用高效液相色谱法测定其中的阿霉素溶解度。最后，MTT法测定ADR-GL对肝癌细胞HepG2的增殖抑制作用，及经流式细胞仪测定CD44+ MDA-MB-231干细胞所占总MDA-MB-231细胞的比例以初步探究其抗肿瘤活性。结果 ADR-GL复合物中阿霉素1525 cm-1红外光谱吸收峰消失，且DSC分析显示ADR-GL在86 ℃处出现一个明显的新的吸收峰，表明阿霉素可能被甘草酸所包裹而以新的形式存在，称之为“ADR-GL分子复合物”。对照组盐酸阿霉素组在pH=7.4的磷酸盐缓冲液中溶解度为0.844±0.011 mmol/L，ADR-GL复合物组与对照组相比溶解度均有显著性差异（P<0.05），其中阿霉素与甘草酸摩尔比为1∶2组溶解度最高，为5.562±0.049 mmol/L，是对照组的6.3倍。此外，与阿霉素对照组相比，ADR-GL分子复合物组对HepG2细胞活力的直接影响及对MDA-MB-231肿瘤干细胞群比负作用二者无显著差异。结论 ADR-GL不降低阿霉素抗肿瘤活性，其可能是一种潜在的安全的具有开发价值的新颖递药系统。

关键词: 阿霉素；甘草酸；分子复合物；抗肿瘤活性

Abstract: Objective To prepare an adriamycin-glycyrrhizin molecular complex (ADR-GL complex) using glycyrrhizin (GL, a component in traditional Chinese drug) as the carrier and assess the solubility and anti-tumor activity of the complex. Methods Dried solid products of ADR-GL complex with different molar ratios of ADR and GL (2∶1, 1∶1, and 1∶2) were prepared by rotary steaming and characterized using FT-IR and DSC. The products were dissolved in pH7.4 phosphate buffer, sonicated overnight, and centrifuged to obtain saturated ADR-GL complex solution, and ADR solubility was determined using high-performance liquid chromatography (HPLC). The cytotoxicity of ADR and ADR-GL complex was evaluated in HepG2 cells by assessing the cell viability using MTT assay. Breast cancer MDA-MB-231 cells were treated with ADR-GL complex and the proportion of CD44 + cells in the total cells was measured by flow cytometry to evaluate the anti- tumor activity of the complex. Results FT-IR spectrum of solid ADR-GL complex did not show the absorption peak of adriamycin at 1525 cm-1 , and an intense absorption peak of ADR-GL occurred at 86 ℃ in DSC, indicating that ADR molecules were encapsulated by GL, the giving rise to the new form of ADR-GL molecular complex. The solubility of ADR in pH7.4 phosphate buffer in the control group was 0.844±0.011 mmol/L, significantly different from that in ADR-GL complex group (P<0.05). The ADR-GL complex with an ADR to GL ratio of 1∶2 showed the highest ADR solubility (5.562 ± 0.049 mmol/L), which was 6.3 times that of the control sample. The ADR-GL complex and ADR showed similar inhibitory effects on HepG2 cells and the negative stemness population of MDA-MB-231 stem cells. Conclusion The ADR-GL complex does not reduce the antitumor activity of ADR and may serve potentially as a safe and novel drug delivery system.

Key words: adriamycin; glycyrrhizin; molecular complex; anti-tumor activity