CN111569071B - 含穿膜肽且光/活性氧刺激脱peg壳的聚乳酸纳米颗粒制备及应用 - Google Patents
含穿膜肽且光/活性氧刺激脱peg壳的聚乳酸纳米颗粒制备及应用 Download PDFInfo
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- CN111569071B CN111569071B CN202010475587.7A CN202010475587A CN111569071B CN 111569071 B CN111569071 B CN 111569071B CN 202010475587 A CN202010475587 A CN 202010475587A CN 111569071 B CN111569071 B CN 111569071B
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Abstract
本发明公开了一种含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系、其制备方法及应用。所述药物体系包含:5.0~5.76重量份的活性药物;85.67~90.41重量份的活性氧响应的光敏嵌段聚合物PEG‑TK‑Ce6‑PLA材料;和4.52~8.57重量份的细胞穿膜肽修饰的聚乳酸TAT‑PLA材料。
Description
技术领域
本发明涉及聚乳酸材料领域,具体涉及一种含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系、其制备方法及应用。
背景技术
近年来癌症纳米医学取得了稳健的发展,该领域的研究与开发经历了指数级增长,并且有许多纳米药物已经被FDA批准用于治疗癌症。纳米颗粒具有许多优点,如能够封装难溶的小分子药物,保护治疗分子,并改变其血液循环和组织分布特征等。目前光响应的纳米药物已经被广泛的应用于研究药物递送系统,光响应体系有比较好时间和空间响应性,并且红光的组织穿透性高,对正常细胞的副作用小。在许多光响应的纳米载体中,可降解的聚合物载体最具有研究前景的。聚合物材料具有性能优越,合成路线简单等许多优点。其中可降解的聚丙交酯(PLA),已成为医学领域最常用的合成生物可降解聚合物之一,并广泛的应用于临床上的药物输送载体。
PEG明显的改善了所修饰药物的溶解性、生物相容性、毒性有以及体内的循环时间等。然而药物的治疗效果却没有发生明显的改变。这是因为PEG层能够使颗粒在体内长循环过程中稳定颗粒,然而在肿瘤细胞内部,由于纳米药物表面的PEG密度高和肿瘤组织内部固有的病理特征,使得纳米药物难以被肿瘤细胞摄取。输送与摄取两难问题便是制约传统纳米药物药效的重要因素之一,也是目前亟待解决的科学问题之一。为了解决这个问题,一大批科研工作者设计了许多策略,使得PEG修饰的纳米颗粒既能在体内长循环,又能在组织部位脱去PEG壳层,以增加肿瘤细胞对纳米药物的摄取。然而许多内源性刺激PEG脱壳的纳米颗粒,都是基于肿瘤组织自身的微环境,然而由于肿瘤组织的异质性,并且不同病人、不同肿瘤以及肿瘤的不同时期的肿瘤微环境可能失效,因此内源刺激响应性的PEG脱壳有很大的局限性。
以解决纳米药物载体的输送-摄取两难问题为研究对象,主动靶向增加纳米药物内化进入肿瘤细胞为目的,设计了具有近红外光诱导断裂的ROS敏感的PEG-TK-Ce6-PLA两亲型嵌段聚合物及细胞穿膜肽(TAT)封端的聚乳酸TAT-PLA两种功能化聚合物。通过两种聚合物组装构建以亲水PEG和TAT为壳层,PLA为负载阿霉素(DOX)疏水核的纳米颗粒。PEG用来遮盖TAT,以保证纳米颗粒在体内的长循环,在近红外的照射下,光敏基团Ce6产生活性氧切断缩硫酮(TK)键导致PEG壳层脱除,使穿膜肽裸露在纳米药物的表面,在其作用下,纳米药物被内化,负载的阿霉素进入肿瘤细胞后和细胞内的DNA相互作用以杀死细胞。
发明内容
为了克服现有技术的不足,本发明的目的是提供一种含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系、其制备方法及应用,基于所述药物体系的总重量,其包含:5.06~5.76重量份的活性药物;85.67~90.41重量份的活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料;和4.52~8.57重量份的细胞穿膜肽修饰的聚乳酸TAT-PLA材料;
其中,所述活性药物为选自阿霉素、紫杉醇、喜树碱中的一种或多种;优选为阿霉素;
所述活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料为
其中,X为45~113的整数,Y为50~100的整数;
所述细胞穿膜肽修饰的聚乳酸TAT-PLA材料为
其中Z为50~150的整数,其YGRKKRRQRRRC为TAT的氨基酸序列。
本发明的一个目的是提供一种含穿膜肽且光、活性氧刺激脱PEG壳的
聚乳酸纳米颗粒制备的方法,该方法包括:
1)制备活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料,其包括如下步骤:
其中,X、Y、的定义同前文;
步骤(1-1)以聚乙二醇单甲醚(其中链节数为45~113)、溴乙酸乙酯为原料,叔丁醇钾为亲核试剂,18-冠醚-6为离子络合剂室温下发生亲核取代反应,生成羧基化的聚乙二醇PEG-COOH;
步骤(1-2)PEG-COOH在二环己基碳二亚胺、N-羟基琥珀酰亚胺存在下活化,并与含有缩硫酮键的二乙胺室温下发生酰胺化反应,生成PEG-TK;
步骤(1-3)Ce6(Chlorine 6)在二环己基碳二亚胺、N-羟基琥珀酰亚胺存在下活化,并与PEG-TK室温下发生酰胺化反应,生成PEG-TK-Ce6;
步骤(1-4)PEG-TK-Ce6在二环己基碳二亚胺、N-羟基琥珀酰亚胺下活化,并与4-氨甲基苯甲醇于溶剂室温下发生酰胺化反应,生成PEG-TK-Ce6-OH;步骤(1-5)以PEG-TK-Ce6-OH为引发剂,1,8-二氮杂二环十一碳-7-烯(DBU)为催化剂,外消旋体丙交酯D,L-LA为单体,室温下发生活性开环聚合,生成ROS敏感的嵌段聚合物PEG-TK-Ce6-PLA;
2)制备聚乳酸TAT-PLA材料,其包括如下步骤:
其中,Z的定义同前文;
步骤(2-1)以带双键小分子HEM为引发剂、外消旋体丙交酯D,L-LA为单体,异辛酸亚锡(Sn(Oct)2)为催化剂发生活性开环聚合,生成带双键的聚合物HEM-PLA;
步骤(2-2)细胞穿模肽TAT和聚合物HEM-PLA在碱性条件下发生点击反应,生成TAT-PLA。
3)制备负载活性药物的纳米药物,其包括如下步骤:
步骤(3-1)首先将活性药物(选自阿霉素(DOX)、紫杉醇、喜树碱中的一种或多种,优选为DOX),PEG-TK-Ce6-PLA,TAT-PLA分别溶于DMSO中,配制成相应溶液(优选,分别为8~15mg/mL,更优选10mg/mL);
步骤(3-2)以活性药物、TAT-PLA、PEG-TK-Ce6-PLA相应溶液的体积比为1:0.5:10~1:1:10。取活性药物溶液、TAT-PLA溶液、PEG-TK-Ce6-PLA溶液加入容器中,搅拌,混合溶液,逐滴加入到高速搅拌的去离子水中,搅拌,而后置于去离子水中透析以除去DMSO溶剂和未负载上的活性药物,最终得到负载活性药物的纳米药物;
优选地,例如,取0.1mL DOX溶液、0.1mL TAT-PLA溶液、1mLPEG-TK-Ce6-PLA溶液加入到圆底烧瓶中,搅拌2h,然后将混合溶液逐滴加入到10mL的高速搅拌的去离子水中,搅拌4h。将溶液转移到截留分子量为14kDa的透析袋中,将透析袋密封,置于5L去离子水中透析,加入磁子缓慢搅拌,以除去DMSO溶剂和未负载上的DOX。每两小时换一次水,透析12小时即得到负载DOX的纳米药物。
优选的,步骤(1-1)所述的聚乙二醇单甲醚,其中X为45~113,进一步优选为113,溴乙酸乙酯,叔丁醇钾,18-冠醚-6的摩尔比为1:20:20:20~1:30:30:30,进一步优选为1:30:30:30。
优选的,步骤(1-1)中所述的反应温度为25℃-35℃,时间为12-24h,进一步优选为25℃,时间为12h。
优选的,步骤(1-2)所述的二环己基碳二亚胺、N-羟基琥珀酰亚胺、小分子缩硫酮单体、PEG-COOH的摩尔比为1.2:1.2:1.5:1~1.5:1.5:2:1,进一步优选为1.2:1.2:1.5:1。
优选的,步骤(1-2)所述的活化时间为6-12h,反应时间为12-24h,进一步优选为活化时间为6h,反应时间为12h。
优选的,步骤(1-3)所述的二环己基碳二亚胺、N-羟基琥珀酰亚胺、二氢卟吩e6、PEG-TK的摩尔比为3.3:3.3:3:1~4:4:4:3:1,进一步优化选为3.3:3.3:3:1。
优选的,步骤(1-4)所述的PEG-TK-Ce6、4-氨甲基苯甲醇的摩尔比为1:2~1:3,进一步优选为1:3。
优选的,步骤(1-4)所述的反应溶剂为甲醇或二甲亚砜,进一步优化选为甲醇。
优选的,步骤(1-5)所述的PEG-TK-Ce6-OH、外消旋体丙交酯、1,8-二氮杂二环十一碳-7-烯的摩尔量之比为1:50:1~1:150:1.5,进一步优化选为1:100:1。
优选的,步骤(1-5)所述的反应温度为20℃-30℃,反应时间为2h-3h,进一步优选为25℃,反应时间为2h。
优选的,步骤(2-1)所述的反应小分子引发剂HEM、异辛酸亚锡、外消旋体丙交酯的摩尔量之比为1:50:0.5~1:100:1,进一步优选为1:50:0.5。
优选的,步骤(2-1)所述的反应的时间为2h-4h,进一步优选为2h。
优选的,步骤(2-2)所述的反应TAT、HEM-PLA的摩尔量之比为1.2:1~1.5:1,进一步优选为1.2:1。
优选的,步骤(2-2)所述的反应时间为12h~24h,进一步优选为24h。
以上所述的制备方法是制得一种活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料和细胞穿膜肽修饰的聚乳酸TAT-PLA材料。
以上所述的活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料和细胞穿膜肽修饰的聚乳酸TAT-PLA材料,作为运输载体负载小分子药物应用于制备载药纳米颗粒。
本发明的另一个目的是提供所含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系在制备载药纳米颗粒中的应用,以及所述药物体系在治疗乳腺癌、肺癌、卵巢癌、胃癌、肝癌的药物中的用途。
这两种材料是通过以下的方式来实现。以聚乙二醇单甲醚、溴乙酸乙酯为原料,叔丁醇钾为亲核试剂,18-冠醚-6为离子络合剂合成羧基化的聚乙二醇PEG-COOH。二环己基碳二亚胺和N-羟基琥珀酰亚胺将PEG-COOH活化,并和含有缩硫酮键的二乙胺反应生成PEG-TK。二环己基碳二亚胺和N-羟基琥珀酰亚胺将Ce6活化,将PEG-TK加入其中,合成PEG-TK-Ce6。然后将PEG-TK-Ce6和4-氨甲基苯甲醇加入到溶剂中,合成PEG-TK-Ce6-4-OH,并以此为引发剂以1,8-二氮杂二环十一碳-7-烯(DBU)为催化剂,外消旋体丙交酯为单体,室温下发生活性开环聚合,合成ROS敏感的嵌段聚合物PEG-TK-Ce6-PLA。
以带双键小分子引发剂HEM,异辛酸亚锡(Sn(Oct)2)为催化剂发生活性开环聚合,合成了带双键的聚合物HEM-PLA。用细胞穿模肽(TAT)在碱性条件下与聚合物HEM-PLA发生点击反应,合成了TAT-PLA。
基于缩硫酮键的活性氧响应的聚乙二醇-聚乳酸结构负载阿霉素的载药纳米颗粒可药物在体内的长循环。并且在到达肿瘤组后,在近红外光的照射下,缩硫酮键裂解,PEG层脱去,TAT暴露在纳米颗粒的表面,TAT能主动携带药物进入细胞,使药物在胞内的含量增加,增加对肿瘤细胞的杀伤能力。
本发明中亲水部分是聚乙二醇,为亲水性聚酯,链节数为113,其相对分子量为5000。
本发明中疏水部分是聚乳酸,它的优点在于①疏水性,通过疏水-疏水相互作用可包载疏水性药物自组装成纳米颗粒;②可生物降解,并且它的最终降解产物不会对生物体有不良影响;③制成的载药纳米颗粒在循环过程中,可避免Ce6泄露;本发明的活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料和TAT修饰的聚乳酸TAT-PLA材料可在水相中自组装形成纳米颗粒并应用于疏水性抗癌药物的运输载体。
本发明得到的活性氧响应的聚乳酸材料具有良好的生物相容性和可降解性。基于活性氧敏感的缩硫酮键构建的载药纳米颗粒,在血液长循环过程中颗粒内的药物几乎不释放,在到达肿瘤部位后,给予近红外光照射,光敏剂产生活性氧,活性氧能够使缩硫酮键断裂,PEG脱去,TAT暴露在纳米颗粒的表面,TAT能主动携带阿霉素进入肿瘤细胞,增加肿瘤部位阿霉素的药物浓度,提高药物的利用率和治疗效果,具有重大的临床应用意义。
附图说明
图1为活性氧响应的光敏嵌段聚合物PEG113-TK-Ce6-PLA材料和细胞穿膜肽修饰的聚乳酸TAT-PLA材料的合成路线。
图2为PEG113-COOH的核磁氢谱图。
图3为PEG113-TK的核磁氢谱图。
图4为PEG113-TK-Ce6的核磁氢谱图。
图5为PEG113-TK-Ce6-OH的核磁氢谱图。
图6为PEG113-TK-Ce-PLA140的核磁氢谱图。
图7为HEM-PLA72的核磁氢谱图。
图8为TAT-PLA72的核磁氢谱图。
图9为PEG113-TK-Ce-PLA200的核磁氢谱图。
图10为HEM-PLA112的核磁氢谱图。
图11为四种载药纳米颗粒在水溶液中的粒径及粒径分布图。
图12为四种载药纳米颗粒的透射电镜图。
图13为PEG113-TK-Ce6-OH的光裂解核磁氢谱图。
图14为PEGNPCe6@DOX纳米颗粒的光脱壳核磁氢谱图。
图15为四种载药纳米颗粒的稳定性图。
图16为四种载药纳米颗粒体外药物释放曲线图。
图17四种载药纳米颗粒对MDA-MB-231细胞的杀伤情况。
具体实施方式
以下结合实例对本发明的具体实施做进一步的说明,但本发明的实施方式不限于此。
聚乙二醇单甲醚PEG113-OH,CAS:9004-74-4,购买自Sigma-Aldrich(上海)贸易有限公司。
二环己基碳二亚胺,CAS:538-75-0,购买于阿拉丁试剂(上海)有限公司。
N-羟基琥珀酰亚胺胺,CAS:228-001-3,购买于阿拉丁试剂(上海)有限公司。
18-冠醚-6,CAS:17455-13-9,购买于阿拉丁试剂(上海)有限公司。
外消旋体丙交酯,CAS:95-96-5,购买于济南大港生物材料有限公司。
溴乙酸乙酯,CAS:105-36-2,购买于阿拉丁试剂(上海)有限公司。
4-氨甲基苯甲醇,CAS:39895-56-2,购买于阿拉丁试剂(上海)有限公司。
二氢卟吩e6,CAS:19660-77-6,购买于百灵威科技有限公司。
1,8-二氮杂二环十一碳-7-烯,CAS:6674-22-2,购买于阿拉丁试剂(上海)有限公司。
异辛酸亚锡,CAS:301-10-0,购买自Sigma-Aldrich(上海)贸易有限公司。
细胞穿膜肽TAT,其氨基酸序列为YGRKKRRQRRRC-SH,购买于杭州中肽生化有限公司。
实施例1、活性氧响应的嵌段聚合物PEG-TK-Ce6-PLA和TAT修饰的聚乳酸TAT-PLA的合成与表征
一、活性氧响应的光敏聚合物PEG113-TK-Ce6-PLA140的合成
(1)将聚乙二醇单甲醚PEG113-OH(20g,0.004mol)、叔丁醇钾(13.44g,0.12mol)和18-冠醚-6(31.68g,0.12mol)溶于甲苯中,在50℃反应30min,冷却至25℃,然后将溴乙酸乙酯(20.04g,0.12mol)加入其中,25℃反
应12h。产物用乙醚沉淀两次,再加入氢氧化钠溶液进行酯水解反应1h,反应结束后,加入盐酸,将溶液的pH调制为3左右。二氯甲烷萃取,无水硫酸钠
干燥,旋蒸即得到PEG113-COOH。
(2)将PEG113-COOH(10g,0.002mol)、二环己基碳二亚胺(0.494g,0.0024mol)、N-羟基琥珀酰亚胺(0.276g,0.0024mol)溶于干燥的二氯甲烷中活化6h,将含有缩硫酮键的二乙胺(0.776g,0.004mol)加入其中反应12h,产物用乙醚沉淀两次,即得到PEG113-TK。
(3)将Ce6(0.1788g,0.0003mol),二环己基碳二亚胺(0.07128g,0.00033mol),N-羟基琥珀酰亚胺(0.03795mg,0.00033mol)放入干燥的二氯甲烷中活化6h,将PEG113-TK(0.5g,0.0001mol)加入其中室温下反应12h。产物用乙醚沉淀两次,得到PEG113-TK-Ce6。
(4)将PEG113-TK-Ce6(0.5g,0.000083mol)和4-氨甲基苯甲醇(0.039g,0.00025mol)加入到甲醇,反应12h,将产物用乙醚沉淀两次,得到PEG113-TK-Ce6-OH。
(5)将PEG113-TK-Ce6-OH(30mg,0.000006mol,1eq)、丙交酯(D,L-LA)(86.4mg,0.0006mol,100eq)加入至含有干燥甲苯的干燥两口瓶中。瓶子上加一个氮气球,用油泵将甲苯抽干。抽干后,将干燥的甲苯加入到两口瓶中,瓶子上加一个氮气球,用双排管将氮气抽排三次,保持瓶内的氮气氛,然后用微量注射器加入1,8-二氮杂二环十一碳-7-烯(1μL,0.000006mol)于25℃下反应2h,加入含有苯甲酸的干燥的甲苯结束反应。用乙醚沉淀两次即得到PEG113-TK-Ce6-PLA140。
二、TAT修饰的聚乳酸TAT-PLA72的合成
(1)将HEM(18mg,0.128mmol,1eq),丙交酯(D,L-LA)(0.92g,6.4mmol,50eq)放入到含有干燥甲苯的干燥两口瓶中,瓶子的翻口塞上加一个氮气球,用油泵将甲苯抽干。抽干后,将干燥的甲苯加入其中,瓶子上加一个氮气球,用双排管将氮气抽排三次,保持瓶内的氮气氛,然后辛酸亚锡加入(2.6mg,0.064mmol,0.5eq)并于105℃下反应2h。加入含有苯甲酸的干燥的甲苯结束反应,用乙醚沉淀两次即得到HEM-PLA72。2)称取HEM-PLA72(165.85mg,0.012mmol,1eq)溶解于二甲亚砜中,然后先往烧瓶中滴入三乙胺调pH为8左右,在氮气氛围下加入TAT(30mg,0.018mmol,1.5eq),室温下搅拌反应24h。反应结束后,进行透析。透析结束后,将透析袋中的液体取出加入到离心管中,冷冻干燥即得到产物TAT-PLA72。
三、活性氧响应的嵌段聚合物PEG113-TK-Ce6-PLA140和TAT修饰的聚乳酸TAT-PLA72的表征
对上述合成材料进行核磁共振氢谱(1H NMR)分析,测定其分子结构:
图2即为聚合物PEG113-COOH的核磁氢谱,c处即新产生的峰,即为键合后新产生的亚甲基峰,同时我们将在不同化学环境下的氢原子对应的峰进行了归属。1H NMR(400MHz,CDCl3,δ):4.13(-CH2-,2H),3.62(-CH2-,452H),3.36(-CH3,3H)。
图3即为PEG113-TK的核磁氢谱图。从核磁氢谱图中可以看出,1.58ppm是TK的的甲基特征峰,同时我们将在不同化学环境下的氢原子对应的峰进行了归属。1H NMR(400MHz,CDCl3,δ):3.36(-CH3,3H),3.62(-CH2-,454H),4.13(-CH2-,2H),2.92(-CH2-,2H),2.75(-CH2-,4H),1.58(-CH3,6H)。
图4即为PEG113-TK-Ce6的核磁氢谱图。从核磁氢谱图中可以看出,PEG-TK-Ce6上出现了Ce6的特征峰,并且Ce6上9.8ppm处羧基的峰消失,说明PEG-TK-Ce6合成成功。
图5即为PEG113-TK-Ce6-OH的核磁氢谱图。从核磁氢谱图中可以看出,PEG113-TK-Ce6-OH在7.25ppm左右出现了4-氨甲基苯甲醇中的苯环峰,说明4-氨甲基苯甲醇成功的接到PEG113-TK-Ce6上,PEG113-TK-Ce6-OH合成成功。
图6即为PEG113-TK-Ce6-PLA140的核磁氢谱图。从核磁氢谱图中可以看出,5.25ppm和1.5ppm即为PEG113-TK-Ce6-PLA140中PLA中次甲基和甲基的特征峰,经过核磁积分可以看出,一共接上了140个链节。
图7即为HEM-PLA72的核磁氢谱图。双键上的峰为7.02ppm,而PLA中次甲基和甲基的特征峰分别为5.25ppm和1.5ppm。
图8即为TAT-PLA72的核磁氢谱图。HEM-PLA72中双键在7.02ppm的峰,在点击反应后得到的产物TAT-PLA72中消失,说明点击反应的成功发生,TAT成功的接到了HEM-PLA72上,成功合成了TAT-PLA72。
实施例2、活性氧响应的嵌段聚合物PEG113-TK-Ce6-PLA200和TAT修饰的聚乳酸TAT-PLA112的合成
一、活性氧响应的光敏聚合物PEG113-TK-Ce6-PLA200的合成
(1)将聚乙二醇单甲醚PEG113-OH(20g,0.004mol)、叔丁醇钾(13.44g,0.12mol)和18-冠醚-6(31.68g,0.12mol)溶于甲苯中,在50℃反应30min,冷却至25℃,然后将溴乙酸乙酯(20.04g,0.12mol)加入其中,25℃反应12h。产物用乙醚沉淀两次,再加入氢氧化钠溶液进行酯水解反应1h,反应结束后,加入盐酸,将溶液的pH调制为3左右。二氯甲烷萃取,无水硫酸钠干燥,旋蒸即得到PEG113-COOH。
(2)将PEG113-COOH(10g,0.002mol)、二环己基碳二亚胺(0.494g,0.0024mol)、N-羟基琥珀酰亚胺(0.276g,0.0024mol)溶于干燥的二氯甲烷中活化6h,将含有缩硫酮键的二乙胺(0.776g,0.004mol)加入其中反应12h,产物用乙醚沉淀两次,即得到PEG113-TK。
(3)将Ce6(0.1788g,0.0003mol),二环己基碳二亚胺(0.07128g,0.00033mol),N-羟基琥珀酰亚胺(0.03795mg,0.00033mol)放入干燥的二氯甲烷中活化6h,将PEG113-TK(0.5g,0.0001mol)加入其中室温下反应12h。产物用乙醚沉淀两次,得到PEG113-TK-Ce6。
(4)将PEG113-TK-Ce6(0.5g,0.000083mol)和4-氨甲基苯甲醇(0.039g,0.00025mol)加入到甲醇,反应12h,将产物用乙醚沉淀两次,得到PEG-TK-Ce6-OH。
(5)将PEG113-TK-Ce6-OH(30mg,0.000006mol,1eq)、丙交酯(D,L-LA)(129.6mg,0.0009mol,150eq)加入至含有干燥甲苯的干燥两口瓶中。瓶子上加一个氮气球,用油泵将甲苯抽干。抽干后,将干燥的甲苯加入到两口瓶中,瓶子上加一个氮气球,用双排管将氮气抽排三次,保持瓶内的氮气氛,然后用微量注射器加入1,8-二氮杂二环十一碳-7-烯(0.75μL,0.0000045mol)于25℃下反应2h,加入含有苯甲酸的干燥的甲苯结束反应。用乙醚沉淀两次即得到PEG113-TK-Ce6-PLA200。
二、TAT修饰的聚乳酸TAT-PLA112的合成
(1)将HEM(18mg,0.128mmol,1eq),丙交酯(D,L-LA)(1.38g,9.6mmol,75eq)放入到含有干燥甲苯的干燥两口瓶中,瓶子的翻口塞上加一个氮气球,用油泵将甲苯抽干。抽干后,将干燥的甲苯加入其中,瓶子上加一个氮气球,用双排管将氮气抽排三次,保持瓶内的氮气氛,然后异辛酸亚锡加入(3.9mg,0.096mmol,0.75eq)并于105℃下反应2h。加入含有苯甲酸的干燥的甲苯结束反应,用乙醚沉淀两次即得到HEM-PLA112。
(2)称取HEM-PLA112(165.85mg,0.012mmol,1eq)溶解于二甲亚砜中,然后先往烧瓶中滴入三乙胺调pH为8左右,在氮气氛围下加入TAT(30mg,0.018mmol,1.5eq),室温下搅拌反应24h。反应结束后,将产物放入到截留分子量为3.5k Da的透析袋中,进行透析。透析结束后,将透析袋中的液体取出加入到离心管中,冷冻干燥即得到产物TAT-PLA112。
图9PEG113-TK-Ce6-PLA200的核磁氢谱图。从核磁氢谱图中可以看出,5.25ppm和1.5ppm即为PEG113-TK-Ce6-PLA200中PLA中次甲基和甲基的特征峰,经过核磁积分可以看出,一共接上了200个链节。
图10即为HEM-PLA112的核磁氢谱图。双键上的峰为7.02ppm,而PLA中次甲基和甲基的特征峰分别为5.25ppm和1.5ppm。
实施例3、活性氧响应的嵌段聚合物PEG113-TK-Ce6-PLA140和TAT修饰的聚乳酸TAT-PLA72纳米颗粒化及应用
一、纳米颗粒的制备
通过纳米沉淀法(Nano precipitation method)制备活性氧响应的PEG脱壳载药纳米颗粒,具体方法如下:
首先将活性药物(选自阿霉素(DOX)、紫杉醇、喜树碱中的一种或多种,优选为DOX),PEG-TK-Ce6-PLA,TAT-PLA分别溶于DMSO中,配制成相应溶液(优选,10mg/mL)。
步骤(3-2)以活性药物、TAT-PLA、PEG-TK-Ce6-PLA相应溶液的体积比为1:0.5:10~1:1:10。取活性药物溶液、TAT-PLA溶液、PEG-TK-Ce6-PLA溶液加入容器中,搅拌,混合溶液,逐滴加入到高速搅拌的去离子水中,搅拌,而后置于去离子水中透析以除去DMSO溶剂和未负载上的活性药物,最终得到负载活性药物的纳米药物;
称取实施例1制备的PEG113-TK-Ce6-PLA140,TAT-PLA72,DOX分别溶于DMSO中,配置成10mg/mL的相应溶液。取0.1mL DOX溶液,0.1mL TAT-PLA72溶液,1mL PEG113-TK-Ce6-PLA140溶液,加入容器中,搅拌,混合溶液。然后将混合溶液逐滴滴加到搅拌的10mL超纯水中,在避光条件下搅拌4h,在超纯水中透析一天。透析结束后,将颗粒溶液用0.45μm过滤器过滤除去未被负载的DOX,得到含TAT-PLA的载药纳米颗粒,记为PEGNPCe6&TAT@DOX。
称取实施例1制备的PEG113-TK-Ce6-PLA140,DOX分别溶于DMSO中,配置成10mg/mL的相应溶液。取0.1mL DOX溶液,1mL PEG113-TK-Ce6-PLA140溶液,加入容器中,搅拌,混合溶液。然后将混合溶液逐滴滴加到搅拌的10mL超纯水中,在避光条件下搅拌4h,在超纯水中透析一天。透析结束后,将颗粒溶液用0.45μm过滤器过滤除去未被负载的DOX,得到不含TAT-PLA的载药纳米颗粒,记为PEGNPCe6@DOX。
利用紫外分光光度计(UV-vis)测试颗粒中DOX的浓度,通过投料的总量减去未包载的DOX的量算得载在纳米颗粒中的药物的含量。
纳米颗粒包载DOX的载药量(drug loading content,DLC)通过以下公式算得:
载药量(%)=颗粒包载DOX的总质量/包载DOX的纳米颗粒的总质量×100%
两种药物PEGNPCe6&TAT@DOX、PEGNPCe6@DOX的载药量分别为5.76%和4.6%。
二、活性氧响应的载药纳米颗粒的完整性和形貌
PEGNPCe6&TAT@DOX、PEGNPCe6@DOX两种纳米颗粒分别用0.2W cm-2光强的660nm激光器分别光照0min和30min,然后进行透射电镜(TransmissionElectron Microscope,TEM)和动态光散射表征。通过图11的DLS图和图12TEM图中可以清楚的观察到,光照前后纳米颗粒的尺寸仍为40nm左右,未发生团聚与崩解,纳米颗粒保持其形貌的完整性,形貌仍为球形。
三、活性氧响应的载药纳米颗粒的PEG脱壳特性
首先以PEG113-TK-Ce6-OH为模型,将其溶于氘代氯仿中,分别用0.2W cm-2光强的660nm激光器光照0min和30min,然后送去进行核磁氢谱的检测。如图13所示,光照30min后,1.58ppm处的TK上的特征峰甲基峰降解,并且在2.15ppm处产生新的丙酮的特征峰。
PEGNPCe6@DOX分别用0.2W cm-2光强的660nm激光器光照0min、10min、20min、30min,光照之后透析24h以除去PEG层,将透析液冻干后进行核磁氢谱的检测。我们以PLA中次甲基峰为标准峰,其积分面积不变,分析PEG中亚甲基峰的积分面积。如图14所示,在光照0min时,标准峰PLA中次甲基峰的氢原子数目为156,特征峰PEG中亚甲基峰的的氢原子数目为456。在光照30min时,标准峰PLA中次甲基峰的氢原子数目为156,特征峰PEG中亚甲基峰的氢原子数目为383。从中可以看出,标准峰PLA的氢原子数不变,而PEG的氢原子却在不断的减少。
四、活性氧响应的载药纳米颗粒的稳定性和药物释放行为
取PEGNPCe6&TAT@DOX(L-)、PEGNPCe6@DOX(L-)、PEGNPCe6&TAT@DOX(L+30min)以及PEGNPCe6@DOX(L+30min),分别取出2mL(DOX含量为100μg)的光照前后的纳米颗粒,加入到两端封口的3.5k Da的透析袋中,取出四根50mL的离心管,里面加入15mL PBS(pH=7.4),将透析袋放入离心管中,将离心管密封好。放置于37℃的恒温震荡水槽中,在2h、4h、8h、12h、24h、36h、48h、60h直至156h,用15mL的等量的PBS溶液换下,将收集的外液测利用荧光光度计测量DOX的含量,然后累加至156h。如图15所示,光照前后的四组纳米药物没有突释行为,光照之后纳米颗粒的DOX的释放有所增加。
取出PEGNPCe6&TAT@DOX(L+30min)、PEGNPCe6&TAT@DOX(L-)、PEGNPCe6@DOX(L+30min)和PEGNPCe6@DOX(L-)0.5mL载药纳米颗粒溶液加入到2mL含有10%胎牛血清(FBS)的高糖培养基(DMEM)中,然后用DLS测量纳米颗粒在72小时内的稳定性,每12小时测一次。如图16所示,载药纳米颗粒的粒径在72小时内没有发生明显变化,光照前后的载药纳米颗粒具有较好的稳定性。
五、活性氧响应载药纳米颗粒的体外细胞实验
我们选取了人乳腺癌细胞系(MDA-MB-231)用于探究活性氧响应纳米颗粒在光照条件下对药物DOX的释放情况。我们选取Free DOX、PEGNPCe6@DOX(L-)、PEGNPCe6&TAT@DOX(L-)、以及光照30min后的纳米颗粒PEGNPCe6&TAT@DOX(L+30min)和PEGNPCe6@DOX(L+30min),其中DOX的浓度分别为2.3μg mL-1,4.6μg mL-1,9.2μg mL-1和18.4μg mL-1,五种药物在37℃共同培养48h,摄取结束后洗去未被摄取的药物或颗粒。最后用MTT法检测各实验组肿瘤细胞活性。从图17中可以看出,未光照的纳米药物,其细胞毒性很低。激光照射后,两组药物均显示出来抑制肿瘤细胞生长的效果,而PEGNPCe6&TAT@DOX(L+30min)组的肿瘤细胞的杀伤效果最好。说明在光照之后载药纳米颗粒PEG的脱壳,使得TAT暴漏在纳米颗粒的表面,在TAT的作用下,纳米药物能主动的进入细胞,从而使得细胞内的DOX的含量增多,从而杀死细胞,提高纳米颗粒的肿瘤杀伤能力。
现有技术制备的两嵌段聚合物PEG113-TK-PLA140,是以PEG113-OH、溴乙酸乙酯为原料,叔丁醇钾为亲核试剂,并将得到的物质通过过阳离子交换柱得到PEG113-COOH。二环己基碳二亚胺和N-羟基琥珀酰亚胺将PEG113-COOH活化,将TK加入其中,合成PEG113-TK。以PEG113-TK为引发剂,异辛酸亚锡Sn(Oct)2为催化剂,外消旋体丙交酯为单体,发生活性开环聚合,合成ROS敏感的嵌段聚合物PEG113-TK-PLA140。并以PEG113-TK-PLA140为材料,通过纳米沉淀法,制备负载化疗药PTX和光敏剂Ce6的纳米颗粒TK-NPCe6&PTX。TK-NPCe6&PTX的粒径约为80nm左右,并且在光照前后纳米药物依然保持完整并且能稳定存在。TK-NPCe6&PTX在0.2W cm-2的660nm的红光光照30min后,PEG约脱壳50%左右。与现有技术相比,PEGNPCe6&TAT@DOX的材料合成更加的简便,并且Ce6接在PEG113-TK上,避免光敏剂Ce6在运输过程中泄露。相对于纳米药物TK-NPCe6&PTX,纳米药物PEGNPCe6&TAT@DOX的尺寸更小,使得纳米颗粒更好地在肿瘤部位聚集。与纳米药物TK-NPCe6&PTX(L+30min)组相比,纳米药物PEGNPCe6&TAT@DOX(L+30min)组即使在少量PEG脱壳的情况下,也达到比较好的杀伤肿瘤的效果。
Claims (6)
1.一种含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系,其中,所述药物体系包含:5.0~5.76重量份的活性药物;85.67~90.41重量份的活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料;和4.52~8.57重量份的细胞穿膜肽修饰的聚乳酸TAT-PLA材料;
其中,所述活性药物为选自阿霉素、紫杉醇、喜树碱中的一种或多种;
所述活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料为:
其中,X为45~113的整数,Y为50~100的整数;
所述细胞穿膜肽修饰的聚乳酸TAT-PLA材料为:
其中,Z为50~150的整数,YGRKKRRQRRRC为TAT的氨基酸序列。
2.根据权利要求1所述的药物体系,其中,所述药物体系包含5.0~5.76重量份的活性药物;85.67~90.41重量份的活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料;和4.52~8.57重量份的细胞穿膜肽修饰的聚乳酸TAT-PLA材料;
其中,所述活性药物为阿霉素;
在所述活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料中,X为45~113的整数,Y为50~100的整数;
在所述细胞穿膜肽修饰的聚乳酸TAT-PLA材料中,Z为50~150的整数。
3.一种制备所述含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系的方法,该方法包括:
1)制备活性氧响应的光敏嵌段聚合物PEG-TK-Ce6-PLA材料,其包括如下步骤:
其中,X、Y的定义如权利要求1所述;
步骤(1-1)以PEG-OH、溴乙酸乙酯为原料,叔丁醇钾为亲核试剂,18-冠醚-6为离子络合剂室温下发生亲核取代反应,生成羧基化的聚乙二醇PEG-COOH;
步骤(1-2)PEG-COOH在二环己基碳二亚胺、N-羟基琥珀酰亚胺存在下活化,并与含有缩硫酮键的二乙胺室温下发生酰胺化反应,生成PEG-TK;
步骤(1-3)Ce6在二环己基碳二亚胺、N-羟基琥珀酰亚胺存在下活化,并与PEG-TK室温下发生酰胺化反应,生成PEG-TK-Ce6;
步骤(1-4)PEG-TK-Ce6在二环己基碳二亚胺、N-羟基琥珀酰亚胺存在下活化,并与4-氨甲基苯甲醇于溶剂中室温下发生酰胺化反应,生成PEG-TK-Ce6-OH;
步骤(1-5)以PEG-TK-Ce6-OH为引发剂,1,8-二氮杂二环十一碳-7-烯(DBU)为催化剂,外消旋体丙交酯D,L-LA为单体,室温下发生活性开环聚合,生成ROS敏感的嵌段聚合物PEG-TK-Ce6-PLA;
2)制备聚乳酸TAT-PLA材料,其包括如下步骤:
其中,Z的定义如权利要求1所述;
步骤(2-1)以带双键小分子HEM为引发剂、外消旋体丙交酯D,L-LA为单体,异辛酸亚锡(Sn(Oct)2)为催化剂发生活性开环聚合,生成末端带双键的聚合物HEM-PLA;
步骤(2-2)细胞穿模肽TAT和聚合物HEM-PLA在碱性条件下发生点击反应,生成TAT-PLA;
3)制备负载活性药物的纳米药物,其包括如下步骤:
步骤(3-1)首先将所述活性药物,PEG-TK-Ce6-PLA,TAT-PLA分别溶于DMSO中,配制成相应溶液;
步骤(3-2)以活性药物、TAT-PLA、PEG-TK-Ce6-PLA相应溶液的质量比为1:0.5:10~1:1:10,取活性药物溶液、TAT-PLA溶液、PEG-TK-Ce6-PLA溶液加入容器中,搅拌,混合溶液,逐滴加入到高速搅拌的去离子水中,搅拌,而后置于去离子水中透析以除去DMSO溶剂和未负载上的活性药物,最终得到负载活性药物的纳米药物。
4.根据权利要求3所述的方法,其中,步骤(1-1)所述的聚乙二醇单甲醚中,其中X为45~113,
步骤(1-1)中,聚乙二醇单甲醚,溴乙酸乙酯,叔丁醇钾,18-冠醚-6的摩尔比为1:20:20:20~1:30:30:30,
步骤(1-1)中所述的反应温度为25℃-35℃,时间为12-24h,
步骤(1-2)所述的二环己基碳二亚胺、N-羟基琥珀酰亚胺、小分子缩硫酮单体、PEG-COOH的摩尔比为1.2:1.2:1.5:1~1.5:1.5:2:1,
步骤(1-2)所述的活化时间为6-12h,反应时间为12-24h,
步骤(1-3)所述的二环己基碳二亚胺、N-羟基琥珀酰亚胺、二氢卟吩e6、PEG-TK的摩尔比为3.3:3.3:3:1~4:4:4:3:1,
步骤(1-4)所述的PEG-TK-Ce6、4-氨甲基苯甲醇的摩尔比为1:2~1:3,
步骤(1-4)所述的反应溶剂为甲醇或二甲亚砜,
步骤(1-5)所述的PEG-TK-Ce6-OH、外消旋体丙交酯、1,8-二氮杂二环十一碳-7-烯的摩尔量之比为1:50:1~1:150:1.5,
步骤(1-5)所述的反应温度为20℃-30℃,反应时间为2h-3h,
步骤(2-1)所述的反应小分子引发剂HEM、异辛酸亚锡、外消旋体丙交酯的摩尔量之比为1:50:0.5~1:100:1,
步骤(2-1)所述的反应的时间为2h-4h,
步骤(2-2)所述的反应TAT、HEM-PLA的摩尔量之比为1.2:1~1.5:1,
步骤(2-2)所述的反应时间为12h~24h。
5.根据权利要求1或2所述的含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系在制备载药纳米颗粒中的应用。
6.根据权利要求1或2所述的含穿膜肽且光/活性氧刺激脱PEG壳的聚乳酸纳米颗粒药物体系在制备治疗乳腺癌、肺癌、卵巢癌、胃癌、肝癌的药物中的用途。
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CN109223729A (zh) * | 2018-09-21 | 2019-01-18 | 华南理工大学 | 一种缩硫酮键键合阿霉素和聚磷酸酯的材料及其制备方法与应用 |
CN109517173A (zh) * | 2018-11-12 | 2019-03-26 | 华南理工大学 | 一种含有缩硫酮键的支化聚醚酰亚胺材料及其制备方法与应用 |
WO2019226651A1 (en) * | 2018-05-22 | 2019-11-28 | Children's Medical Center Corporation | Nanoparticles for treatment of choroidal neovascularization and other indications |
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CN109223729A (zh) * | 2018-09-21 | 2019-01-18 | 华南理工大学 | 一种缩硫酮键键合阿霉素和聚磷酸酯的材料及其制备方法与应用 |
CN109517173A (zh) * | 2018-11-12 | 2019-03-26 | 华南理工大学 | 一种含有缩硫酮键的支化聚醚酰亚胺材料及其制备方法与应用 |
Non-Patent Citations (3)
Title |
---|
On-demand PEGylation and dePEGylation of PLA-based nanocarriers via amphiphilic mPEG-TK-Ce6 for nanoenabled cancer chemotherapy;Yueqiang Zhu, et al.;《Theranostics》;20191022;第9卷(第26期);第8312-8320页及支持材料 * |
ROS-sensitive thioketal-linked polyphosphoester-doxorubicin conjugate for precise phototriggered locoregional chemotherapy;Pei Pei,et al.;《Biomaterials》;20181011;第188卷;第74-82页 * |
红光调控ROS的敏感聚合物纳米载体治疗效果的脱壳和增强肿瘤研究;李杰;《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ⅰ辑》;20190115(第01期);第B016-1301页 * |
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