甘草次酸修饰姜黄素阳离子脂质体介入治疗肝癌的作用及机制研究

After confirmed diagnosis of liver cancer , about 80% of patients can only be treated by non-surgical treatments .The interventional therapy is the first choice of the non-surgical treatments .But when interventional therapy for liver cancer , the adopted antitumor drugs are usually toxic. and their liver-targeting poor , causing long-term effect subpar ,adverse effect comparatively large . So we developed the non-toxic drugs and targeting preparation of Curcumin cationic Liposomes modified by Glycyrrhetinic Acid , and a patent for invention was applied (No. 201110324751.5) for the preparation already . After non-toxic antitumor Curcumin was prepared cationic Liposome , the Curcumin could be transferred into cell nuclei of hepatoma ,which its therapeutic index could be increased, the bottleneck problem of its instability and the hardly soluble in water could be resolved . Glycyrrhetinic Acid possess carboxylate anion ,there are a large number of high specific and high active receptors on the surface of hepatocyte , after modified with Glycyrrhetinic Acid ,the whole Liposome showed electronegative or neutral , so the problems of the pulmonary embolism caused by cationic Liposomes and the hepatocyte-targeting poor would be solved . The interventional therapy of the liposome preparation was security and high activity ..Through experimental methods of cellular pharmacology in vitro and anti-tumor pharmacology of animal liver cancer model with interventional therapy , by means of technologies of tumor immunology and molecular biology ,we will deeply explore the effects and mechanism of the interventional therapy for liver cancer with the liposome preparation , laying foundations for its further development and utilization .

肝癌确诊后约80%的患者只能采用非手术疗法，介入治疗是非手术疗法中的首选方法，但肝癌介入治疗通常采用毒性抗癌药，且靶向性差，致使远期疗效欠佳，毒副反应较大。我们研究开发了甘草次酸修饰姜黄素阳离子脂质体介入治疗肝癌的无毒药物靶向治疗制剂，已申请发明专利（201110324751.5）。无毒抗癌药姜黄素制成阳离子脂质体，可将其导入肝癌细胞核内，提高了其治疗指数，解决了其不稳定和难溶于水的应用"瓶颈问题"。甘草次酸具带负电的羧酸根，且肝细胞表面存在大量高特异性、高活性的甘草次酸受体，甘草次酸修饰姜黄素阳离子脂质体，使脂质体整体带负电或中性，可解决阳离子脂质体在体内易致肺栓塞和肝实质细胞靶向性差的问题。该制剂介入治疗肝癌，安全高效。本项目通过体外细胞药理学和动物肝癌模型介入治疗抗肿瘤药理学等实验方法，运用肿瘤免疫学、分子生物学等技术深入研究该制剂介入治疗肝癌的作用及机制，为进一步开发应用奠定基础。

项目背景：肝癌确诊后约80%的患者只能采用非手术疗法，介入治疗是非手术疗法中的首选方法，但肝癌介入治疗通常采用毒性抗癌药，且全身分布，肝靶向性差，致使远期疗效欠佳，毒副反应较大。为此，我们开发了无毒药物肝靶向给药治疗系统----甘草次酸修饰姜黄素阳离子脂质体。.主要研究内容：1.甘草次酸十八胺两亲复合物的合成；2. 甘草次酸修饰姜黄素阳离子脂质体制备工艺优化； 3.甘草次酸修饰姜黄素阳离子脂质体、甘草次酸联合姜黄素、姜黄素联合索拉非尼、姜黄素联合阿霉抗肝癌细胞株HepG-2的作用；4.甘草次酸修饰姜黄素阳离子脂质体瘤内介入及尾静脉注射治疗H22肝癌移植瘤、大鼠尾静脉注射治疗肝Walker-256移植瘤的作用 ..重要结果：1.两亲性导向分子材料甘草次酸十八胺复合物通过静电方式结合而成，合成方式简单、流程短、成本低；2.甘草次酸修饰姜黄素阳离子脂质体通过乙醇注入法制备，工艺简单，可大量制备，肝靶向作用明显，同时解决了姜黄素难溶于水及不稳定的问题；3. 甘草次酸与姜黄素联合应用、姜黄素联合索拉非尼、甘草次酸修饰姜黄素阳离子脂质体均能明显增强抗HepG-2肝癌细胞作用；4.甘草次酸修饰姜黄素阳离子脂质体瘤内介入及静脉注射治疗H22肝癌移植瘤、静脉注射大鼠肝Walker-256移植瘤的作用显著增强。..关键数据：甘草次酸修饰的姜黄素阳离子脂质体处方：甘草次酸十八胺化合物26mg，卵磷脂200mg，姜黄素4mg，蒸馏水20g。脂质体粒径圆整，分散均匀。脂质体粒径194±0.25nm，PdI0.214±0.02；电位+31.9±0.31mv。脂质体包封率=98.26%。活体成像实验结果表明从0.5—6h，只有肝、脾有荧光，心、肺、肾没有荧光。小鼠及大鼠种植瘤实验结果表明姜黄素脂质体低剂量组（5mg/kg）肿瘤抑制率（24.33%）与姜黄素组（20mg/kg）肿瘤抑制率（23.91%）相当，说明姜黄素制备成姜黄素阳离子脂质体后，抗肿瘤作用大大增强。..科学意义：我们开发的甘草次酸修饰姜黄素阳离子脂质体无毒药物靶向制剂，可提高姜黄素治疗指数，并解决其难溶于水和不稳定的“瓶颈问题”；较好的解决了目前阳离子脂质体在动物体内易致肺栓塞和肝实质细胞靶向性差的问题；为肝癌的治疗提供了新的思路和工具。