CN115531316A - 一种奈达铂脂质体组合物及其制备方法 - Google Patents
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Abstract
本发明提供一种奈达铂脂质体组合物及制备方法，采用薄膜分散法制备奈达铂脂质体，脂质体组合物中的主要辅料为磷脂和胆固醇，其特征在于所选择的磷脂种类应为氢化卵磷脂或者人工合成磷脂，不能使用常规的天然不饱和卵磷脂。
Description
技术领域
本发明涉及药物制剂技术领域，具体涉及一种奈达铂脂质体组合物及制备方法。
背景技术
肺癌是世界范围内最常见的癌症之一，其中非小细胞肺癌占比最高，非小细胞肺癌是一种致命疾病，其五年生存率仅为18％。根据肺癌指南，铂类药物的化疗是治疗非小细胞肺癌的一线方案。铂类药物主要通过结合细胞内DNA靶点而发挥细胞毒性作用，但是，铂类药物对肿瘤部位和正常组织都显示出一定的细胞毒性。一些经典的铂类药物，如顺铂、卡铂、奥沙利铂、奈达铂等已被批准用于临床，其中奈达铂是顺铂的衍生物，比顺铂具有更大的水溶性和更快的排泄速率。奈达铂降低了与顺铂相关的肾毒性和胃肠道毒性，然而，奈达铂的剂量限制毒性为骨髓抑制，主要表现为白细胞减少、贫血和血小板减少等，这些副作用限制了其临床应用。因此，为了降低奈达铂的全身毒性，提高其抗肿瘤疗效，有必要开发新的奈达铂制剂。
脂质体具有生物相容性好、非免疫原性等特点，可将药物与外界环境分离，保护药物不被降解。此外，脂质体可以通过聚乙二醇修饰来避免网内皮系统的摄取，能延长药物在体内的作用时间，可被动靶向至肿瘤部位，提高药物的治疗效果和减轻毒副作用。迄今为止，尚无铂类药物脂质体制剂上市，本发明提供一种奈达铂脂质体组合物及其制备方法，药效学研究显示其具有提高药物治疗效果的作用。
发明内容
本发明提供了一种奈达铂脂质体组合物及制备方法，采用薄膜分散法，首先将磷脂和胆固醇按一定比例溶解于有机溶剂中，然后使用旋转蒸发仪减压挥干有机溶剂，直到形成薄膜，再加入奈达铂溶液水化，透析除去游离药物，通过高压均质机或超声的方法减小粒径，最后挤出过膜得到脂质体制剂。
所述磷脂为氢化卵磷脂(HSPC)或者人工合成磷脂(如DSPC、DSPE、DPPC等)，优选价格低廉的氢化大豆磷脂和DSPE-PEG2000的混合物，其比例范围在1：1～20：1。
所述磷脂不能使用常规的天然不饱和卵磷脂(如大豆卵磷脂和蛋黄卵磷脂)。
所述奈达铂脂质体组合物中，磷脂和胆固醇的比例范围在1：1～4：1。
所述用于水化薄膜的奈达铂溶液浓度为1～10mg/mL。
所述有机溶剂为乙醇、丙酮和氯仿，优选乙醇。
本发明提供的奈达铂脂质体组合物为静脉注射用，动物实验表明，该制剂的安全性和市售制剂相当，但其肿瘤抑制作用比溶液剂显著提高，能显著提高药物治疗的有效性，具有较强的市场应用前景。
附图说明
为了使本发明的目的、技术方案和有益效果更加清楚，本发明提供如下附图进行说明：
图1为不同种类的磷脂与奈达铂原料药混合于水中后的液相色谱图。
图2为奈达铂脂质体的外观及粒径分布图
图3为奈达铂脂质体的透射电镜图
图4为奈达铂脂质体的体外释放曲线图
图5小鼠静脉注射PBS对照、奈达铂溶液剂和脂质体制剂后肿瘤体积变化趋势图
图6小鼠静脉注射PBS对照、奈达铂溶液剂和脂质体制剂后的肿瘤图
图7各给药组的血常规和肝肾功指标检测结果
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明，以使本领域的技术人员可以更好的理解本发明并能予以实施，但所举实施例不作为对本发明的限定。
实施例1
目前尚无关于奈达铂与磷脂相容性的报道，本发明将不同种类的磷脂与奈达铂原料药按1:1的比例混合并分散于水中后，采用HPLC分析方法分析奈达铂的降解情况。如图1所示，将奈达铂(NDP)分别与大豆卵磷脂(SPC)和蛋黄卵磷脂(EPC)混合后，在相对保留时间为9.3min左右出现了一个明显的降解峰，并且奈达铂的药物峰明显减小，说明大豆卵磷脂和蛋黄卵磷脂会促使奈达铂降解。但是，奈达铂与氢化大豆磷脂(HSPC)或人工合成磷脂(DSPC、DPPC)混合后，没有出现明显的药物降解峰，表明奈达铂与氢化大豆磷脂或者人工合成磷脂具有很好的相容性，本发明优选价格低廉的氢化大豆磷脂制备脂质体，为了提高脂质体的制剂稳定性和延长体内作用时间，本发明的脂质体组合物中还加入了辅料DSPE-PEG2000。
实施例2
奈达铂脂质体采用薄膜分散法制备，其中，药物和HSPC的比例为1：4，HSPC和胆固醇的比例为2：1，DSPE-mPEG2000的比例为20％。首先将HSPC和DSPE-mPEG2000溶解于氯仿中，然后置于旋转蒸发仪中，在45℃、150转/分的条件下减压蒸发溶剂，直到形成薄膜。加入奈达铂溶液，60℃水化6小时，脂质体悬浮液在200W条件下超声4分钟，再过孔径为220nm的聚碳酸酯膜，透析除游离药物后得奈达铂脂质体。测得脂质体得平均粒径为134.73±2.80nm，PDI值为0.19±0.01，可见粒径较小，分布均匀(图1)。由图2可见，脂质体为球形，图3表明所制备的脂质体具有明显的缓释作用，在体外的释放时间可达8天。
实施例3
建立了肺癌小鼠模型，当肿瘤长到100～150mm3时，将小鼠随机分为3组(n＝5)，按2mg/kg的给药剂量，分别于第1、5天静脉注射PBS对照、奈达铂溶液剂和奈达铂脂质体制剂，治疗过程中监测小鼠体重和肿瘤体积变化，当肿瘤体积达到约1000～1500mm3时处死小鼠，实验结果如图5和图6所示，奈达铂脂质体给药组的肿瘤体积显著低于奈达铂溶液剂组，可见奈达铂脂质体的疗效如溶液剂相比有明显提高。图7可见，奈达铂脂质体给药后，与奈达铂溶液剂相比，其血常规指标和肝肾功指标的测定值没有显著性差异，证明了该制剂的安全性和市售制剂相当，但其有效性明显提高，具有较强的市场应用前景。
Claims (5)
1.一种奈达铂脂质体组合物，采用薄膜分散法制备，先将磷脂和胆固醇按一定比例溶解于有机溶剂中，然后使用旋转蒸发仪减压挥干有机溶剂，直到形成薄膜，再加入奈达铂溶液水化，透析除去游离药物，通过高压均质机或超声的方法减小粒径，最后挤出过膜得到脂质体制剂。
2.根据权利要求1所述的奈达铂脂质体组合物，其特征在于：所选择的磷脂种类应为氢化卵磷脂或者人工合成磷脂，不能使用常规的天然不饱和卵磷脂。
3.根据权利要求1所述的奈达铂脂质体组合物，其特征在于：磷脂和胆固醇的比例范围在1：1～4：1。
4.根据权利要求1所述的薄膜分散法，其特征在于：所述用于水化薄膜的奈达铂溶液浓度为1～10mg/mL。
5.根据权利要求1所述的薄膜分散法，其特征在于：所述有机溶剂为乙醇、丙酮和氯仿。
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