CN113384709B - 一种葡聚糖-原卟啉前药纳米胶束的制备和应用 - Google Patents
一种葡聚糖-原卟啉前药纳米胶束的制备和应用 Download PDFInfo
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Abstract
本发明公开了一种葡聚糖‑原卟啉前药纳米胶束的制备和应用,制备方法包括以下步骤:(1)氨基化原卟啉(PpIX‑NH2)的制备;(2)醛基化葡聚糖(DEX‑CHO)的制备;(3)DEX‑CHO‑PpIX‑NH2及其DEX‑PpIX胶束的制备。所制备的胶束粒径稳定、分散均一、较高药物上载量、低毒副作用、良好的生物安全性、良好的药物控释等优势,从根本上改善了原卟啉的水溶性差的问题。葡聚糖对原卟啉的修饰,大大降低光敏剂对正常细胞的毒副作用,同时,小分子前药在癌细胞中可控释放,致使原卟啉在肿瘤部位有效富集,通过激光照射,杀死癌细胞实现精确诊疗,为小分子光疗前药递送系统的发展提供一种新的策略和系统性的理论支撑。
Description
技术领域
本发明涉及高分子化学药物及部分生物领域,具体涉及到一种葡聚糖-原卟啉前药纳米胶束的制备方法和应用。
背景技术
癌症是世界范围内的主要公共卫生问题之一,已经成为一个威胁我国人民健康的主要杀手。世界卫生组织国际癌症研究机构 (IARC)近日发布了2020年全球最新癌症负担数据。据统计预测,2020 年全球将有癌症新发病例约1929 万例,死亡约996 万例,其中中国新发癌症病例约457万,占据全球的23.7%,死亡人数约300万,占全球癌症死亡人数的约30%。故此,寻觅一个新方法新策略去提高癌症的治愈率是大势所趋。针对肿瘤治疗,目前已有手术治疗、放射治疗、化学治疗、光动力治疗、免疫治疗等疗法在临床中取得显著进展。
光动力疗法(PDT)是一种具有高度选择性的微创癌症治疗方法。它在光照时通过使用光敏剂将能量转移到周围的分子氧以生成活性氧(ROS),主要是单线态氧(1O2)用于诱导细胞凋亡或坏死和组织破坏,以抑制肿瘤生长。研究表明,PDT 的侵入性明显低于手术,此外,PDT 不会产生任何与放射治疗相关的长期副作用。因此,与传统癌症治疗相比,PDT具有巨大的优势。
在各种光敏剂中,原卟啉(PpIX)是 PDT 中最常用的卟啉之一。原卟啉是一种内源性光敏剂(PS),一般是由是通过给予5-氨基乙酰丙酸(ALA)等前体获得。使用5-ALA的一个缺点是给药后,向PpIX的生物转化时间约为4-6小时,这意味着PDT的延迟阶段,这对癌症治疗造成较大影响。如果直接进行PpIX给药,PpIX本质上较差的水溶性使得直接静脉内给药成为一个挑战。因此,通过优化递送方案来提高这种光敏剂的PDT效率已成为广泛追求的目标。
本发明着手于改善原卟啉的水不溶性,通过酰腙键将原卟啉与葡聚糖缀合,利用肿瘤微环境呈酸性这一特性,展现出葡聚糖-原卟啉前药较高的生物相容性及精准可控释放的能力,从而进一步改善光动力治疗的缺陷。该前药必将推动光疗前药的发展,为光动力治疗的低毒副作用、高精准治疗以及与其他治疗癌症的手段共联提供一定的参考,为小分子光疗前药递送系统的发展提供一种新的策略和系统性的理论支撑。
发明内容
本发明所解决的技术问题提供一种光敏剂和糖类共同递送的小分子前药的制备方法,从根本上解决了光敏剂水溶性差及不稳定的问题。研究在肿瘤酸性环境下,小分子前药的基本表征、药物释放、细胞吞噬、细胞毒性等,旨在提供一种葡聚糖-原卟啉前药的制备方法及其在小分子前药光疗癌症方面的应用。。
本发明的技术方案具体如下:
一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于,包括以下步骤:
(1)整个反应过程在氩气环境中进行,氩气作为保护气,防止氧气或氢气的存在对反应造成影响;同时,整个制备过程中因光敏剂原卟啉的存在,应进行避光处理;将一定量的原卟啉PpIX溶于无水二氯甲烷DCM,得到原卟啉溶液,随后在冰浴下加入一定量的N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐EDC、N-羟基琥珀酰亚胺NHS不断搅拌使其溶解,溶解后继续冰浴搅拌30min,随后加入一定量的肼,室温下反应6h;反应结束后,用去离子水反复萃取3次,得到PpIX-NH2;
(2)将一定量葡聚糖溶解于二甲基亚砜DMSO和N-甲基吡咯烷酮NMP混合溶液中,超声溶解,得到葡聚糖溶液;所述的混合溶液中DMSO与NMP的体积比为1:1;
(3)整个反应过程在氩气且避光的环境中进行,将一定量氯铬酸吡啶盐PCC溶于DMSO,待PCC完全溶解后,快速搅拌下用注射器将步骤(2)的葡聚糖溶液逐滴滴入,室温下反应6h;反应结束后,加入乙醚反复洗涤产物2次,这时会有部分沉淀出现,除去上清,加入乙醚继续洗涤,洗去副产物三乙胺盐;加入二氯甲烷CH2Cl2洗涤2次,除去上清,洗去溶剂DMSO以及部分未反应完全的PCC;最后再次用乙醚洗涤,去上清,待沉淀中的乙醚挥发完全后,真空干燥箱干燥24h,得到DEX-CHO;
(4)整个反应过程在氩气且避光的环境中进行,将一定量的PpIX-NH2和DEX-CHO加入二甲基亚砜DMSO与N,N-二甲基亚砜DMF的混合溶液中,搅拌下加入一定量三乙胺,室温条件下反应24h;反应结束,以甲醇为透析液,分子量为3500的透析袋透析约24h,取出产物,进行旋蒸;旋蒸至还剩些许液体时,加入乙醚沉淀,离心去除上清液,真空干燥24h,得到DEX-CHO-PpIX-NH2;所述混合溶液中DMSO与DMF体积比为2:3;
(5)将一定量DEX-CHO-PpIX-NH2溶于DMF,随后将其用移液枪慢慢滴入快速搅拌状态下的去离子水中,得到混合溶液;搅拌30min,转移至分子量3500的透析袋透析约24h,透析液为去离子水,得到DEX-PpIX胶束。
进一步的,步骤(1)中,原卟啉溶液的摩尔浓度范围为0.01~0.06mol·L-1;所述EDC与NHS摩尔比为1:1且EDC与NHS摩尔浓度范围为0.04~0.12mol·L-1。
进一步的,步骤(2)中,葡聚糖溶液的摩尔浓度范围为0.01~0.03mol· L-1。
进一步的,步骤(3)中,PCC的摩尔浓度范围为0.15~0.40 mol·L-1;洗涤产物过程中,产物与乙醚的体积比范围为1:1~1:2;产物与二氯甲烷的体积比范围为1:1~1:2。
进一步的,步骤(4)中,PpIX-NH2质量浓度范围为7~9 mg·mL-1;DEX-CHO质量浓度范围为35-40 mg·mL-1;三乙胺的体积30-60μL;透析过程中产物与甲醇的体积比例范围为1:150~1:300。
进一步的,步骤(5)DEX-CHO-PpIX-NH2溶于DMF中得到的质量浓度范围为0.1~0.2mg·mL-1;混合溶液的质量浓度范围为0.5~0.8 mg·mL-1;透析过程中胶束与去离子水的体积比例范围为1:200~1:300。
本发明的主要优点在于:
1.针对光动力治疗中光敏剂存在水溶性差的问题,以起关键作用的光敏剂原卟啉为例,葡聚糖修饰,增强其稳定性,从根本上解决其水溶性差的问题,为其静脉给药提供了极大的便利,成功优化递送方案以改善水溶性较差的光敏剂的光动力治疗效率。
2.在酸性肿瘤微环境中,葡聚糖与原卟啉连接的酰腙键断裂,释放出光敏剂原卟啉,可用于荧光成像和光声成像等,实现药物的可控释放及精确诊疗。由葡聚糖修饰后,通过细胞毒性可看出,胶束在正常细胞中几乎无毒性,证明其对正常组织的低毒副作用、较高生物安全性。而在激光照射下用于PDT治疗,继续生成单线态氧来杀死癌细胞,实现肿瘤的多模态协同治疗。
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:
图1为实施例1中葡聚糖-原卟啉前药的合成示意图。
图2为实施例1中葡聚糖-原卟啉前药的核磁示意图。
图3为实施例1中葡聚糖-原卟啉前药纳米胶束的紫外光谱图。
图4为实施例1中葡聚糖-原卟啉前药纳米胶束的荧光光谱图。
图5为实施例1中葡聚糖-原卟啉前药纳米胶束的透射电镜图TEM和动态光散射粒径分布图DLS。
图6为实施例1中葡聚糖-原卟啉前药纳米胶束的体外药物释放示意图。
图7为实施例1中葡聚糖-原卟啉前药纳米胶束对4T 1细胞和L929成纤维细胞的毒性对比图。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。
实施例1 制备一种葡聚糖-原卟啉前药
一种葡聚糖-原卟啉前药的总合成示意图如图1所示,主要包括以下步骤:
(1)整个反应过程在氩气环境中进行,氩气作为保护气,防止氧气或氢气的存在对反应造成影响;同时,整个制备过程中因光敏剂原卟啉的存在,应进行避光处理;将原卟啉(PpIX)(57mg,0.1mmol)溶于5mL无水二氯甲烷DCM,得到原卟啉溶液,随后在冰浴下加入N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸EDC(75mg,0.4mmol)、N-羟基琥珀酰亚胺NHS(46mg,0.4mmol)不断搅拌使其溶解,溶解后继续冰浴搅拌30min,随后加入肼(1.2mmol),室温下反应6h;反应结束后,用去离子水反复萃取3次,得到PpIX-NH2;
(2)将葡聚糖(1.5g,0.3mmol)溶解于10mL二甲基亚砜DMSO和N-甲基吡咯烷酮NMP混合溶液中,超声溶解,得到葡聚糖溶液;所述的混合溶液中DMSO与NMP的体积比为1:1;
(3)整个反应过程在氩气且避光的环境中进行,将氯铬酸吡啶盐PCC(1g,4.6mmol)溶于10mL二甲基亚砜DMSO,待PCC完全溶解后,快速搅拌下用注射器将步骤(2)的葡聚糖溶液逐滴滴入,室温下反应6h;反应结束后,加入乙醚反复洗涤产物2次,每次乙醚用量为25mL,这时会有部分沉淀出现,除去上清,加入乙醚继续洗涤,洗去副产物三乙胺盐;加入二氯甲烷CH2Cl2洗涤2次,每次二氯甲烷用量为25mL,除去上清,洗去溶剂DMSO以及部分未反应完全的PCC,最后再次用乙醚洗涤2次,每次乙醚用量为25mL,去上清,待沉淀中的乙醚挥发完全后,真空干燥箱干燥24h,得到DEX-CHO;
(4)整个反应过程在氩气且避光的环境中进行,将40mg PpIX-NH2和192mg DEX-CHO加入5mL二甲基亚砜DMSO与N,N-二甲基亚砜DMF的混合溶液中,搅拌下加入40μL三乙胺,室温条件下反应24h;反应结束,以甲醇为透析液,透析液500mL,分子量为3500的透析袋透析约24h,取出产物,进行旋蒸;旋蒸至还剩2mL液体时,加入乙醚沉淀,离心去除上清液,真空干燥24h,得到DEX-CHO-PpIX-NH2;所述混合溶液中DMSO与DMF体积比为2:3;
(5)将5mg DEX-CHO-PpIX-NH2溶于0.6mL DMF,随后将其用移液枪慢慢滴入快速搅拌状态下的3mL去离子水中,搅拌30min,转移至分子量3500的透析袋透析24h,透析液为500mL去离子水,得到DEX-PpIX胶束。图3紫外光谱图、图4荧光光谱图有效证明胶束DEX-PpIX的成功合成。DP胶束的粒径分布及尺寸大小如图5所示。图6为DEX-PpIX纳米胶束的体外释放示意图,可知药物在体外模拟肿瘤酸性环境中较好释放效果。图7为DEX-PpIX胶束4T1肿瘤细胞及L929成纤维细胞的毒性对比图,可以看出胶束对正常细胞几乎无毒副作用,对癌细胞有较大杀伤力,突出了该胶束的PDT治疗效果。
最后说明的是,以上优选实施例仅用以说明本发明的技术方案而非限制,尽管通过上述优选实施例已经对本发明进行了详细的描述,但本领域技术人员应当理解,可以在形式上和细节上对其作出各种各样的改变,而不偏离本发明权利要求书所限定的范围。
Claims (6)
1.一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于,包括以下步骤:
(1)整个反应过程在氩气环境中进行,氩气作为保护气,防止氧气或氢气的存在对反应造成影响;同时,整个制备过程中因光敏剂原卟啉的存在,应进行避光处理;将一定量的原卟啉PpIX溶于无水二氯甲烷DCM,得到原卟啉溶液,随后在冰浴下加入一定量的N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐EDC、N-羟基琥珀酰亚胺NHS不断搅拌使其溶解,溶解后继续冰浴搅拌30min,随后加入一定量的肼,室温下反应6h;反应结束后,用去离子水反复萃取3次,得到PpIX-NH2;
(2)将一定量葡聚糖溶解于二甲基亚砜DMSO和N-甲基吡咯烷酮NMP混合溶液中,超声溶解,得到葡聚糖溶液;所述的混合溶液中DMSO与NMP的体积比为1:1;
(3)整个反应过程在氩气且避光的环境中进行,将一定量氯铬酸吡啶盐PCC溶于DMSO,待PCC完全溶解后,快速搅拌下用注射器将步骤(2)的葡聚糖溶液逐滴滴入,室温下反应6h;反应结束后,加入乙醚反复洗涤产物2次,这时会有部分沉淀出现,除去上清,加入乙醚继续洗涤,洗去副产物三乙胺盐;加入二氯甲烷CH2Cl2洗涤2次,除去上清,洗去溶剂DMSO以及部分未反应完全的PCC;最后再次用乙醚洗涤,去上清,待沉淀中的乙醚挥发完全后,真空干燥箱干燥24h,得到DEX-CHO;
(4)整个反应过程在氩气且避光的环境中进行,将一定量的PpIX-NH2和DEX-CHO加入二甲基亚砜DMSO与N,N-二甲基亚砜DMF的混合溶液中,搅拌下加入一定量三乙胺,室温条件下反应24h;反应结束,以甲醇为透析液,分子量为3500的透析袋透析约24h,取出产物,进行旋蒸;旋蒸至还剩些许液体时,加入乙醚沉淀,离心去除上清液,真空干燥24h,得到DEX-CHO-PpIX-NH2;所述混合溶液中DMSO与DMF体积比为2:3;
(5)将一定量DEX-CHO-PpIX-NH2溶于DMF,随后将其用移液枪慢慢滴入快速搅拌状态下的去离子水中,得到混合溶液,搅拌30min,转移至分子量3500的透析袋透析约24h,透析液为去离子水,得到DEX-PpIX胶束。
2.根据权利要求1所述的一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于:所述步骤(1)中, 原卟啉溶液的摩尔浓度范围为0.01~0.06mol·L-1;所述EDC与NHS摩尔比为1:1且EDC与NHS摩尔浓度范围为0.04~0.12mol·L-1。
3.根据权利要求1所述的一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于:所述步骤(2)中,葡聚糖的摩尔浓度范围为0.01~0.03mol· L-1。
4.根据权利要求1所述的一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于:所述步骤(3)中,PCC的摩尔浓度范围为0.15~0.40 mol·L-1;洗涤产物过程中,产物与乙醚的体积比范围为1:1~1:2;产物与二氯甲烷的体积比范围为1:1~1:2。
5.根据权利要求1所述的一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于:所述步骤(4)中, PpIX-NH2质量浓度范围为7~9 mg·mL-1;DEX-CHO质量浓度范围为35-40mg·mL-1;三乙胺的体积30-60μL;透析过程中产物与甲醇的体积比例范围为1:150~1:300。
6.根据权利要求1所述的一种葡聚糖-原卟啉前药纳米胶束的制备方法,其特征在于:步骤(5)DEX-CHO-PpIX-NH2溶于DMF中得到的质量浓度范围为0.1~0.2mg·mL-1;混合溶液的质量浓度范围为0.5~0.8 mg·mL-1;透析过程中胶束与去离子水的体积比例范围为1:200~1:300。
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Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20080095182A (ko) * | 2007-04-23 | 2008-10-28 | 한국과학기술연구원 | 고분자 유도체-광감작제 복합체를 이용한 새로운 광역학치료제 |
CN103041405A (zh) * | 2012-12-26 | 2013-04-17 | 深圳先进技术研究院 | 诊疗一体化载药聚合物及其制备方法 |
CN103330938A (zh) * | 2013-07-11 | 2013-10-02 | 中国科学院化学研究所 | 肿瘤靶向性光敏剂及其制备方法与应用 |
CN108774301A (zh) * | 2018-07-11 | 2018-11-09 | 西南大学 | 一类基于葡聚糖的酸响应聚合物药物的制备方法及其应用 |
CN109069472A (zh) * | 2016-05-06 | 2018-12-21 | 般财团法人生物动力学研究所 | 含有高分子化药物的医药组合物 |
CN110041475A (zh) * | 2019-04-11 | 2019-07-23 | 中国科学技术大学 | 一种两亲性嵌段共聚物、其壳交联胶束及制备方法和应用 |
CN111643673A (zh) * | 2020-07-08 | 2020-09-11 | 福建医科大学孟超肝胆医院(福州市传染病医院) | 一种同时包载光敏剂和蛋白质的肿瘤靶向纳米药物及其应用 |
CN112618727A (zh) * | 2021-01-08 | 2021-04-09 | 中国药科大学 | 一种增强乏氧肿瘤光动力治疗的制剂及其制备方法和应用 |
-
2021
- 2021-06-28 CN CN202110716005.4A patent/CN113384709B/zh active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20080095182A (ko) * | 2007-04-23 | 2008-10-28 | 한국과학기술연구원 | 고분자 유도체-광감작제 복합체를 이용한 새로운 광역학치료제 |
CN103041405A (zh) * | 2012-12-26 | 2013-04-17 | 深圳先进技术研究院 | 诊疗一体化载药聚合物及其制备方法 |
CN103330938A (zh) * | 2013-07-11 | 2013-10-02 | 中国科学院化学研究所 | 肿瘤靶向性光敏剂及其制备方法与应用 |
CN109069472A (zh) * | 2016-05-06 | 2018-12-21 | 般财团法人生物动力学研究所 | 含有高分子化药物的医药组合物 |
CN108774301A (zh) * | 2018-07-11 | 2018-11-09 | 西南大学 | 一类基于葡聚糖的酸响应聚合物药物的制备方法及其应用 |
CN110041475A (zh) * | 2019-04-11 | 2019-07-23 | 中国科学技术大学 | 一种两亲性嵌段共聚物、其壳交联胶束及制备方法和应用 |
CN111643673A (zh) * | 2020-07-08 | 2020-09-11 | 福建医科大学孟超肝胆医院(福州市传染病医院) | 一种同时包载光敏剂和蛋白质的肿瘤靶向纳米药物及其应用 |
CN112618727A (zh) * | 2021-01-08 | 2021-04-09 | 中国药科大学 | 一种增强乏氧肿瘤光动力治疗的制剂及其制备方法和应用 |
Non-Patent Citations (8)
Title |
---|
A Step-by-Step Multiple Stimuli-Responsive Nanoplatform for Enhancing Combined Chemo-Photodynamic Therapy;Yi Wang et al;《Advanced Materials》;20170127;全文 * |
Light-Activated ROS-Responsive Nanoplatform Codelivering Apatinib and Doxorubicin for Enhanced Chemo-Photodynamic Therapy of Multidrug-Resistant Tumors;Xiao Wei;《ACS Appl. Mater. Interfaces》;20180508;全文 * |
Recent advances in innovative strategies for enhanced cancer photodynamic therapy;Tingting Hu et al;《Theranostics》;20210115;第11卷(第7期);全文 * |
Tumor Microenvironment-Responsive Nanomaterials as Targeted Delivery Carriers for Photodynamic Anticancer Therapy;Houhe Liu et al;《Frontiers in Chemistry》;20200929;第8卷;全文 * |
Tumor-targeting core-shell structured nanoparticles for drug procedural controlled release and cancer sonodynamic combined therapy;Lei Wang et al;《J Control Release》;20170717;第28卷(第286期);全文 * |
多重刺激响应型聚合物纳米颗粒在癌症治疗中的研究;王毅;《中国博士学位论文全文数据库 (工程科技Ⅰ辑)》;20181015;全文 * |
胞内示踪的光动-化疗协同治疗 pH敏感纳米载药系统的构建和评价;杜琛;《中国优秀博硕士学位论文全文数据库(硕士)(工程科技Ⅰ辑))》;20210115;全文 * |
还原响应型高分子药物递送系统的构建及其在增强癌症治疗中的应用;王亚君;《中国优秀博硕士学位论文全文数据库(硕士)(工程科技Ⅰ辑))》;20200115;全文 * |
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