CN107281161A - 一种药物纳米制剂及其制备方法 - Google Patents
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Abstract
本发明公开了一种药物纳米制剂及其制备方法。该药物纳米制剂包括纳米载体，所述纳米载体为由亲水外链和疏水内核组成的核壳结构所述亲水外链为水溶性聚合物或水溶性多糖；所述疏水内核为聚酰胺‑胺，其表面吸附有siRNA，内部包埋有抗肿瘤药物和疏水性探针；所述亲水外链与疏水内核通过偶氮苯‑4，4‑二羧酸连接。本发明首次采用偶氮苯‑4，4‑二羧酸(AZO)连接甲氧基聚乙二醇胺(PEG)与聚酰胺‑胺(PAMAM)制备了一种低氧响应性的纳米载体，可用于共载化疗药物和siRNA；该纳米制剂不仅在肿瘤低氧微环境刺激下粒径减小，穿透性强；而且制剂工艺简单，对肿瘤细胞的毒性增加。
Description
技术领域
本发明涉及纳米载药制剂的制备领域，尤其涉及一种药物纳米制剂及其制备方法。
背景技术
癌症是引起人类死亡的第二大原因，肿瘤发病率和死亡率居高不下，使其成为医疗领域的重大挑战之一。化疗仍然是目前最主要的肿瘤治疗手段，但传统的标准化疗往往会产生多种不可避免的副反应及系统毒性。纳米传递系统的出现提高了药物对肿瘤部位的靶向性，使得药物更多得聚集于肿瘤部位，从而大大降低了对其他正常组织器官的毒副作用。目前已有多个抗肿瘤纳米制剂问世，比如Doxil和Abraxane。
现有技术中，关于抗肿瘤纳米制剂的研究较多。利用肿瘤微环境内低氧、偏酸、高压等特点设计纳米给药系统是当前的热点。如Zhu等合成的MMP2敏感脂质体可携载多柔吡星(DOX)治疗乳腺癌。Cheng等将MMP2敏感穿透肽(R9GPLGLAGE8,ACPP)与原卟啉结合形成胶束用于光动力学治疗。在成像方面，Yaping Wang等制备的高效低毒纳米探针可用于探测MMP2高表达的细胞和肿瘤。
然而，在肿瘤部位，细胞外基质更加致密、血管异常化、组织间隙液压增高，受这些因素影响，纳米粒子通常只能被动地聚集在疏漏的肿瘤血管周围，而不能渗透整个肿瘤组织。同时，绝大部分实体瘤都存在的低氧特性造成肿瘤细胞的转录因子HIF1alpha表达水平过高，这是导致肿瘤产生化学耐药的重要原因之一。
目前的传统纳米制剂并不能很好地解决上述这两个问题。因此，构建一个具有肿瘤靶向性并可穿透进入肿瘤内部克服肿瘤低氧耐药问题的药物递送系统十分重要。
发明内容
本发明的目的在于针对现有纳米制剂难以渗透进入肿瘤内部及克服肿瘤耐药问题的不足，提供了一种药物纳米制剂及其应用，该药物纳米制剂是一种可共载化疗药物和siRNA的纳米载体，其在肿瘤低氧微环境刺激下粒径减小进而促进穿透，进入肿瘤内部，提高对肿瘤细胞的毒性。
一种药物纳米制剂，包括纳米载体，所述纳米载体为由亲水外链和疏水内核组成的核壳结构，所述亲水外链为水溶性聚合物或水溶性多糖；所述疏水内核为聚酰胺-胺，内部包埋有抗肿瘤药物；所述亲水外链与疏水内核通过偶氮苯-4，4-二羧酸连接。
上述聚酰胺-胺是一种表面带大量正电的树状大分子物质，能够与低氧刺激响应性小分子偶氮苯-4，4-二羧酸通过酰胺键连接；其表面正电荷能够吸附带负电的核酸。
本发明制备的药物纳米制剂(简称：纳米制剂)中的纳米载体为亲水性外链包裹疏水性内核的壳核式结构，疏水性内核可大量包埋疏水性药物，而带正电性的疏水内核易吸附负电的核酸。纳米载体利用亲水性外壳在体内长循环并通过EPR效应聚集于肿瘤部位后，在肿瘤低氧还原的微环境下，偶氮苯-4，4-二羧酸的偶氮双键断裂，纳米粒子的亲水性外壳脱去，暴露出携载药物和核酸的表面阳离子的小粒径树状大分子，渗透进入肿瘤组织内部细胞，并在溶酶体内发生质子化效应，逃逸进入细胞浆，同时释放抗肿瘤药物和核酸。其中，功能性核酸可用于抑制耐药基因的表达，从而联合药物杀伤低氧条件下的耐药细胞，同时利用该纳米载体携载功能性探针可有效实时反馈药效。
所述的亲水外链可以为水溶性聚合物，如甲氧基聚乙二醇胺(mPEG-NH2)；或者水溶性多糖，如透明质酸(Hyaluronan,HA)，肝素，葡聚糖，海藻酸钠等。
作为优选，所述亲水外链的分子量为800Da～1000000Da。若选用的亲水外链的分子量较大，最终形成的纳米制剂的粒径会增加，因此要选用适合的分子量，以控制纳米制剂的粒径。
作为优选，所述亲水外链为聚乙二醇类衍生物。更优选，所述亲水外链为为甲氧基聚乙二醇胺。甲氧基聚乙二醇胺生物相容性好，能减小纳米制剂的毒性，同时可有效增加纳米制剂体内循环时间，提高纳米制剂的肿瘤靶向性。
进一步地，纳米载体中，所述偶氮苯-4，4-二羧酸和甲氧基聚乙二醇胺的摩尔比为1:1；聚酰胺-胺与偶氮苯-4，4-二羧酸的摩尔比为1：(10～50)。
作为优选，所述甲氧基聚乙二醇胺的分子量为800～8000Da，此时纳米粒的粒径在100～300nm，利于纳米粒蓄积在肿瘤部位。
所述的抗肿瘤药物为疏水性药物，选自盐酸阿霉素、博来霉素、佐柔比星、表柔比星、柔红霉素、喜树碱和紫杉醇中的一种。
聚酰胺-胺是高度枝化的单分散大分子，以乙二胺为核，与丙烯酸甲酯通过Michael加成和酰胺化反应得到，具有高度的几何对称性。低代数的树状大分子是开放的结构，但随着代数的增长，分子量与表面反应基团约增加为前一代的两倍。4代及4代以上聚酰胺-胺的整体结构逐渐呈现球形，内部存在空腔，可以包裹药物分子。
作为优选，所述的疏水内壳为5代聚乙二胺树枝状聚合物(polyamidoaminedendrimer,PAMAM)，抗肿瘤药物为盐酸阿霉素；抗肿瘤药物通过旋蒸法包埋于PAMAM的内部空腔中；所述纳米载体与盐酸阿霉素的质量比为1:(0.01～0.5)。
进一步地，所述疏水内壳表面吸附有siRNA，所述siRNA为低氧诱导因子-1αsiRNA。(HIF1 alpha siRNA)。绝大部分实体瘤都存在的低氧特性造成肿瘤细胞的转录因子HIF1alpha表达水平过高，这是导致肿瘤产生化学耐药的重要原因之一。通过HIF1 alpha siRNA可沉默HIF1 alpha的表达，克服低氧耐药的问题。
作为优选，所述纳米载体与低氧诱导因子-1αsiRNA的摩尔比为(1～500):1。通过凝胶电泳实验与纳米制剂表面电位测量实验发现，该摩尔比下siRNA可完全吸附于纳米制剂表面，且易于与药物共载。
进一步地，所述疏水内壳内部包埋有疏水性探针，所述疏水性探针为2',7'-二氯荧光黄双乙酸(DCFH-DA)；DCFH-DA已被证明是一种成熟的活性氧探针，易于标记于细胞内。
作为优选，所述纳米载体与2',7'-二氯荧光黄双乙酸的摩尔比为(1～2000):1；该比例下，纳米载体易于携载2',7'-二氯荧光黄双乙酸。
本发明还提供了所述纳米载体的制备方法，包括以下步骤：
1)偶氮苯-4，4-二羧酸(AZO)用吡啶溶解后，加入羧基活化剂，活化AZO的羧基，得到活化的AZO溶液；
2)甲氧基聚乙二醇胺(mPEG-NH2)溶于吡啶后，加入活化的AZO溶液中，滴加三乙胺，室温搅拌过夜；得到PEG-AZO溶液；
3)真空旋转蒸发除去PEG-AZO溶液中的吡啶溶液，纯水复溶，得PEG-AZO水溶液；纯化后，加入羧基活化剂，活化AZO的羧基，得到活化的PEG-AZO水溶液。
4)将活化的PEG-AZO水溶液缓慢加入PAMAM水溶液中，用1mol/L NaOH调溶液pH至8，室温搅拌过夜；透析纯化后，冷冻干燥得PEG-AZO-PAMAM(PAP)纳米载体。
上述纳米载体的合成路径如图1所示。
本发明还提供了所述药物纳米制剂的制备方法，包括以下步骤：
1)将DOX·HCl溶解于甲醇：丙酮(1:1，v/v)混合溶剂中，加入三乙胺，碱化；再向碱化后的DOX溶液中加入PEG-AZO-PAMAM甲醇：丙酮(1:1，v/v)混合溶液，避光搅拌72h；
2)50℃旋转蒸发，蒸干溶剂后，以PBS7.4水溶液复溶，置于透析袋(MWCO 3500)中，PBS7.4水溶液介质中避光透析4h，得PAP+DOX制剂，避光4℃保存；
3)将PAP+DOX制剂与siRNA混合于PBS7.4溶液中,37℃，100rpm恒温震荡30min，得PAP+DOX+siRNA药物纳米制剂。
上述药物纳米制剂的具体作用机理为：利用低氧响应性连接分子AZO连接PAMAM和PEG，合成壳核式纳米载体；同时共载DOX和HIF1 alpha siRNA；利用PEG体内长循环的特点和纳米粒的EPR效应聚集到肿瘤组织，在肿瘤低氧还原微环境下发生响应性解离，脱去表面的PEG，暴露粒径较小的PAMAM携载表面的siRNA和内部DOX，渗透进入肿瘤组织内部细胞，并在溶酶体内发生质子化效应，逃逸进入细胞浆，同时释放DOX和siRNA；其中HIF1 alphasiRNA可抑制HIF1的表达，从而联合DOX杀伤低氧条件下的耐药细胞；同时利用该纳米载体携载ROS探针如DCFH-DA可实时反馈药效。
所述药物纳米制剂在抗肿瘤细胞的应用中效果甚佳；本发明以A549肿瘤细胞或MCF-7肿瘤细胞为例很好地验证了此点。
与现有技术相比，本发明具有以下有益效果：
(1)本发明首次采用偶氮苯-4，4-二羧酸(AZO)连接甲氧基聚乙二醇胺(PEG)聚乙二醇与聚酰胺-胺(PAMAM)制备了一种低氧响应性的纳米载体，可共载化疗药物和siRNA，得到药物纳米制剂；该药物纳米制剂不仅具有在肿瘤低氧微环境刺激下粒径减小，增加穿透性的优点；而且制剂工艺简单，与传统抗肿瘤药物单体的溶液相比，该制剂对肿瘤细胞的毒性大大增加。
(2)本发明制备的药物纳米制剂可携载HIF1 alpha siRNA可有效克服肿瘤低氧耐药的问题。
(3)本发明制备的药物纳米制剂可进一步联合ROS探针实现实时反馈药效的功能；在诊疗一体化方面有很好的应用前景。
附图说明
图1为制备20％PEG2K-AZO-PAMAM纳米载体的合成路径示意图；
图2为制备的mPEG(分子量2000),PAMAM(5代)，mPEG-AZO,20％PEG2K-AZO-PAMAM纳米载体的核磁共振图；
图3为制备的20％PEG2K-AZO-PAMAM，10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM纳米载体的表征结果图；
图4为制备的20％PEG2K-AZO-PAMAM，10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM纳米载体的核磁共振图；
图5为制备的20％PEG2K-AZO-PAMAM纳米载体的粒径分布图和表面电位图；
图6为制备的20％PEG2K-AZO-PAMAM纳米载体在常氧及低氧处理后的透射电镜图；
图7为制备的20％PEG2K-AZO-PAMAM+DOX+siRNA药物纳米制剂的透射电镜图；
图8为制备的20％PEG2K-AZO-PAMAM+DOX+siRNA药物纳米制剂在低氧和常氧条件下作用于MCF-7乳腺癌细胞4h后的细胞摄取量流式检测图；
图9为20％PEG2K-AZO-PAMAM+DOX纳米制剂、20％PEG2K-AZO-PAMAM+DOX低氧处理纳米制剂和DOX水溶液对A549细胞球的穿透性实验激光共聚焦图片；
图10为20％PEG2K-AZO-PAMAM+DOX+siRNA纳米制剂低氧下对MCF-7细胞的毒性实验；
图11为20％PEG2K-AZO-PAMAM+DOX纳米制剂与DCFH-DA探针共同作用于A549肿瘤细胞球4小时后的激光共聚焦图片。
具体实施方式
下列实施例中采用的药物、试剂、细胞株和动物具体如下：
敏感性人乳腺癌细胞(MCF-7)；SD大鼠，由浙江大学实验动物中心提供。盐酸多柔比星(Doxorubicin，DOX·HCl，浙江海正药业股份有限公司)；聚酰胺-胺树状大分子(PAMAM，G5.0，Sigma-Aldrich)；偶氮苯-4，4-二羧酸(Azobenzene-4,4’-dicarboxylicAcid,梯希爱上海化工工业有限公司)；吡啶(Pyridine，阿拉丁试剂有限公司)；甲氧基聚乙二醇胺(Methoxypolyethylene glycd amine，H2NCH2CH2(OCH2CH2)n OCH3,MW＝2000，5000阿拉丁试剂有限公司)；连二亚硫酸钠(sodium hyposulfite，Na2S2O4，阿拉丁试剂有限公司)；4-2甲氨基吡啶(4-DMAP,阿拉丁试剂有限公司)；三乙胺(分析纯，国药集团化学试剂有限公司)；二甲基甲酰胺(DMF，Acros Organics)；1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDC·HCl，Sigma-Aldrich)；N-羟基琥珀酰亚胺(NHS，Sigma-Aldrich)；磷酸盐缓冲液(phosphate buffered saline,PBS)；胎牛血清、青-链霉素、不含EDTA的胰酶、0.25％EDTA-胰酶(GIBCO)；R1640培养液(吉诺生物医药技术有限公司)；3-(4，5-二甲基噻唑-2)-2，5-二苯基四氮唑溴盐(MTT，上海生工生物工程有限公司)；二甲基亚砜(DMSO，分析纯)(杭州双林化工试剂厂)；透析袋(截留量为8000-14,000Da)(Spectrum Laboratories,USA)；透析袋(截留量为3500Da)(Spectrum Laboratories,USA)；0.22μm滤膜微孔滤膜(上海新亚净化器件厂)。
实施例1
一、制备20％PEG2K-AZO-PAMAM(20％PEG接枝率，PEG分子量2000)纳米载体
1)取5mg AZO，超声溶于3ml吡啶中，加入4.37mg EDC·HCl，2.56mg NHS和0.23mg4-DMAP，用1mol/L HCl调溶液pH至5，室温搅拌2小时，使AZO的羧基活化。
2)取37mg mPEG-NH2(MW＝2000)，溶于1ml吡啶中；磁力搅拌下，将mPEG吡啶溶液缓慢加入AZO溶液中，滴加5μl三乙胺；室温搅拌过夜。
3)50℃，真空旋转蒸发除去吡啶溶液，得橙黄色薄膜产物。加入10ml纯水复溶，3000rpm，5min离心除去游离的AZO沉淀，得mPEG-AZO水溶液。加入4.37mg EDC·HCl，2.56mgNHS，用1mol/L HCl调溶液pH至5，室温搅拌2小时，活化AZO的羧基。
4)量取100μl 20.5％PAMAM(G5.0)甲醇溶液，置于60℃烘箱中烘干，加5ml去离子水溶解。磁力搅拌下，将mPEG-AZO水溶液缓慢加入PAMAM溶液中，用1mol/L NaOH调溶液pH至8。室温搅拌过夜。
5)取出溶液，置于透析袋(MWCO 8000～14,000)中，超纯水溶液介质中透析10h，透析过程中换水。
6)将溶液从透析袋中取出，即得20％PEG2K-AZO-PAMAM制剂。4℃保存备用。将部分溶液冻干，待用。
二、制备20％PEG2K-AZO-PAMAM+DOX纳米制剂
1)称取5mg DOX·HCl，溶解于2.5ml甲醇：丙酮(1:1，v/v)混合溶剂中，加入10μl三乙胺，碱化。按DOX与PAMAM质量比1:20缓慢将DOX溶液加入实施例1中20％PEG2K-AZO-PAMAM甲醇：丙酮(1:1，v/v)混合溶液避光搅拌72h。
2)50℃旋转蒸发，蒸干溶剂后以PBS7.4水溶液复溶，置于透析袋(MWCO 3500)中，PBS7.4水溶液中避光透析4h得20％PEG2K-AZO-PAMAM+DOX制剂，避光4℃保存。
三、制备20％PEG2K-AZO-PAMAM+DOX+siRNA纳米制剂
将20％PEG2K-AZO-PAMAM+DOX纳米制剂与siRNA按摩尔比为10:1混合于PBS7.4溶液中,37℃，100rpm恒温震荡30min，得20％PEG2K-AZO-PAMAM+DOX+siRNA纳米制剂。
实施例2
本实施例除mPEG-NH2的分子量为5000，投药量为92.5mg外，其余步骤与实施例1相同，制备得20％PEG5K-AZO-PAMAM(20％PEG接枝率，PEG分子量5000)。
实施例3
本实施例除20.5％PAMAM(G5.0)的投药量为200μl外，其余步骤与实施例1相同，制备得10％PEG2K-AZO-PAMAM(10％PEG接枝率，PEG分子量2000)。
实施例4
本实施例除20.5％PAMAM(G5.0)的投药量为200μl外，其余步骤与实施例2相同，制备得10％PEG5K-AZO-PAMAM(10％PEG接枝率，PEG分子量5000)。
实施例5表征与性能分析
对上述实施例1～4的多功能纳米制剂进行形态、核磁共振分析、对实施例1的纳米制剂进行低氧响应性、细胞毒性、渗透性进行分析：
1、对20％PEG2K-AZO-PAMAM，10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM纳米载体进行核磁共振分析
将中间产物PEG-AZO,空白20％PEG2K-AZO-PAMAM制剂用冷冻干燥仪冻干，取PAMAM甲醇溶液60℃烘干。取处理后的PEG-AZO，20％PEG2K-AZO-PAMAM制剂冻干粉及PAMAM，及粉末mPEG-NH2(MW＝2000)，均用D2O作为溶剂溶解，进行1H-NMR分析。参见图2。
结果表明：PAMAM的1H-NMR(D2O，δ)：2.28(br，j)，2.50(br，l)，2.58～2.8(br，k+h)，3.10～3.30(br，i)。PEG的H来源于其结构上的-CH2-CH2-O-的结构，其化学位移在3.4至3.8ppm之间。其中3.60ppm为PEG的亚甲基特征峰，3.42ppm为PEG的甲基特征峰。δ在2.2～3.3处为PAMAM上C-H的特征峰，AZO苯环上的H的化学位移在7.8～8.2。
由此可判断，中间产物PEG与AZO发生了结合，且最终与PAMAM生成了20％PEG2K-AZO-PAMAM。
1单位mPEG-NH2有176个H。1单位PAMAM表面有128个NH2,有2528个H，j处质子峰有632个H。根据b,c,d,e处质子峰与j处质子峰的曲线下面积，计算mPEG2000与PAMAM的摩尔比，得出13mPEG-2k-PAMAM中mPEG与PAMAM的摩尔比为11.5:1(接枝率为11.5％)，计算得分子量为54931。与投料比13相比，反应的产率为88.46％。考虑到1H NMR的积分存在一定误差，可以认为该聚合物按照投料量进行反应，偶联mPEG个数为13。同理，对10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM纳米载体进行计算分析。参见图3和4。
2、制剂粒径、表面电位测定
采用Malvern Zetasizer Nano ZS90系列激光粒径分析仪，测定1mol/L 20％PEG2K-AZO-PAMAM，10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM制剂的粒径和表面Zeta电位。10％PEG2K-AZO-PAMAM，20％PEG5K-AZO-PAMAM，10％PEG5K-AZO-PAMAM制剂结果记录于图3。20％PEG2K-AZO-PAMAM结果参见图5。
3、对20％PEG2K-AZO-PAMAM进行形态观察和低氧敏感性验证
PEG2000-AZO-PAMAM制剂中AZO在低氧环境中易被还原，氮氮双键断裂生成苯胺。PAMAM表面修饰的mPEG断裂，暴露出粒径约为10nm的PAMAM，制剂粒径变小。
Na2S2O4作为常用的硫酸盐类还原剂可迅速消耗水溶液中的氧气，被用于模拟低氧还原环境。
取2份20％PEG2K-AZO-PAMAM空白载体水溶液(5mg/ml)，一份加入现配Na2S2O4水溶液(Na2S2O4终浓度100μM)，涡旋混匀，37℃孵育30min；一份不处理作为空白对照。再将处理制剂与空白制剂置于透析袋(MWCO 1000)中，纯水溶液介质中透析2h，取出；分别加至专用铜网上，在透射电子显微镜下分别观察粒子的大小和形态。参见图6。
20％PEG2K-AZO-PAMAM+DOX+siRNA(5mg/ml)水溶液加至专用铜网上，在透射电子显微镜下分别观察粒子的大小和形态。参见图5。
结论：20％PEG2K-AZO-PAMAM电镜下形态为类球形，粒径均一为200nm左右；低氧处理后粒径减小为20nm左右。所得的纳米制剂在低氧刺激下粒径减小。20％PEG2K-AZO-PAMAM+DOX+siRNA电镜下形态为类球形，粒径较为均一。
4、20％PEG2K-AZO-PAMAM+DOX+siRNA纳米制剂低氧和常氧条件下作用于MCF-7乳腺癌细胞4h后的细胞摄取量
取对数生长期的MCF-7细胞以1ml/孔(5*105个细胞/孔)接种于48孔板中，在37℃，5％CO2条件下培养24小时后，弃去培养液，给予20％PEG2K-AZO-PAMAM+DOX+FAM-siRNA(DOX浓度5μg/ml，FAM-siRNA浓度100μmol/L)250μl，37℃孵箱培养4h，弃去培养液，用冰PBS迅速漂洗3次，加入200μl胰酶消化细胞。光镜下观察细胞变圆、细胞连接稀疏后，加入完全培养液1ml中和胰酶，用枪头轻轻吹打，使细胞脱壁、分散，形成细胞悬液，1000r/min离心5min，弃上清液。500μl细胞培养用PBS重悬细胞于流式管中，进行流式分析。
结论：如图8所述，合成的纳米制剂具有良好的入胞能力，尤其在低氧刺激下，纳米制剂表现出更好的入胞能力。
5、DOX水溶液、20％PEG2K-AZO-PAMAM+DOX纳米制剂和20％PEG2K-AZO-PAMAM+DOX低氧处理纳米制剂对A549细胞球的穿透性实验
12孔板中培养出约500μm的A549肿瘤细胞球。终浓度4μg/ml的DOX,和相同DOX浓度的PAP+DOX,低氧预处理4h的PAP+DOX，37℃在HBSS溶液中与肿瘤球共孵育4h，冰PBS溶液清洗肿瘤球，最后用甲醛固定肿瘤球。共聚焦荧光显微镜下每隔5μm拍摄Z轴荧光图像。参见图9。
结论：游离DOX溶液组DOX吸附在肿瘤球表面，而PAP+DOX制剂拥有较好的穿透能力，尤其在低氧预处理后。
6、20％PEG2K-AZO-PAMAM+DOX+siRNA纳米制剂对MCF-7细胞的毒性实验
MCF-7(1×104个/孔)接种于96孔培养板，常氧组过夜培养24小时后加入不同浓度的阿霉素溶液，常氧孵箱中培养48小时。低氧组过夜培养18小时后，转移至低氧孵箱(1％氧气浓度)中培养6小时。分别加入不同阿霉素浓度的本发明纳米制剂PAP+DOX、PAP+DOX+HIF-1αsiRNA和阿霉素水溶液作为实验组，空白对照组加入等体积的细胞培养液，低氧孵箱中继续培养48h之后，每孔加入100μl 4℃预冷的TCA溶液(10％，w/v)固定细胞。静置5min移入4℃冰箱中固定1h，取出用去离子水冲洗5遍，室温晾干。待96孔板室温下晾干后，每孔加入0.4％(w/v)的SRB染液(1％的乙酸配制)60μl，染色15min后倒掉染液，用1％(v/v)乙酸冲洗4次，去除未结合的染料，室温晾干。用100μl非缓冲Tris-base碱液(10mM，pH＝10.5)溶解与细胞蛋白结合的染料，水平摇床上振荡20min，采用酶标仪515nm处测定光吸收值A值。
按以下公式计算细胞毒性：
细胞毒性(％)＝(1-实验组A值/空白对照组A值)×100％
结果表明：低氧下阿霉素对肿瘤细胞的毒性减弱，而在相同浓度下，本发明的纳米制剂PAP+DOX对肿瘤细胞的毒性大于阿霉素水溶液，携载HIF-1αsiRNA后药效最好。参见图10。
7、20％PEG2K-AZO-PAMAM+DOX纳米制剂与DCFH-DA探针作用于A549肿瘤细胞球
12孔板中培养约500μm的A549肿瘤细胞球。加入20％PEG2K-AZO-PAMAM+DOX+DCFH-DA溶液，37℃在HBSS溶液中与肿瘤球共孵育4h。冰PBS溶液清洗肿瘤球，最后用甲醛固定肿瘤球。共聚焦荧光显微镜下每隔5μm拍摄Z轴荧光图像。参见图11。
结论：纳米制剂联合DCFH-DA探针较好反映出肿瘤细胞收药物刺激后的ROS水平情况。
Claims (10)
1.一种药物纳米制剂，包括纳米载体，所述纳米载体为由亲水外链和疏水内核组成的核壳结构，其特征在于，所述亲水外链为水溶性聚合物或水溶性多糖；所述疏水内核为聚酰胺-胺，内部包埋有抗肿瘤药物；所述亲水外链与疏水内核通过偶氮苯-4，4-二羧酸连接。
2.如权利要求1所述的药物纳米制剂，其特征在于，所述亲水外链为甲氧基聚乙二醇胺。
3.如权利要求2所述的药物纳米制剂，其特征在于，纳米载体中，所述偶氮苯-4，4-二羧酸和甲氧基聚乙二醇胺的摩尔比为1:1；聚酰胺-胺与偶氮苯-4，4-二羧酸的摩尔比为1：(10～50)。
4.如权利要求2所述的药物纳米制剂，其特征在于，所述甲氧基聚乙二醇胺的分子量为800～8000Da。
5.如权利要求2所述的药物纳米制剂，其特征在于，所述疏水内壳为5代聚乙二胺树枝状聚合物；所述抗肿瘤药物为盐酸阿霉素。
6.如权利要求2所述的药物纳米制剂，其特征在于，所述疏水内壳表面吸附有siRNA，所述siRNA为低氧诱导因子-1αsiRNA。
7.如权利要求6所述的药物纳米制剂，其特征在于，所述纳米载体与低氧诱导因子-1αsiRNA的摩尔比为(1～500):1。
8.如权利要求2所述的药物纳米制剂，其特征在于，所述疏水内壳内部包埋有疏水性探针，所述疏水性探针为2',7'-二氯荧光黄双乙酸。
9.如权利要求8所述的药物纳米制剂，其特征在于，所述纳米载体与2',7'-二氯荧光黄双乙酸的摩尔比为(1～2000):1。
10.一种如权利要求2～9任一所述药物纳米制剂的制备方法，其特征在于，所述纳米载体的制备，包括以下步骤：
(1)将羧基活化的偶氮苯-4，4-二羧酸溶液与甲氧基聚乙二醇胺、三乙胺混合后，搅拌过夜，除去溶剂、纯水复溶，得到甲氧基聚乙二醇胺-偶氮苯-4，4-二羧酸水溶液；
(2)向羧基活化的甲氧基聚乙二醇胺-偶氮苯-4，4-二羧酸水溶液中加入聚酰胺-胺水溶液，搅拌过夜、纯化、冷冻干燥后，得到所述纳米载体。
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| CN111743861A (zh) * | 2020-07-15 | 2020-10-09 | 南通大学 | 靶向三阴性乳腺癌的低氧响应手性药物胶束及其制备方法 |
| CN111773184A (zh) * | 2020-07-14 | 2020-10-16 | 山东大学 | 一种乏氧响应脂质体及在制备抗肿瘤药物中的应用 |
| CN111821419A (zh) * | 2020-07-31 | 2020-10-27 | 浙江大学 | 一种自组装多肽纳米载体及其制备方法和应用 |
| CN112402625A (zh) * | 2020-11-19 | 2021-02-26 | 健进制药有限公司 | 负载光敏剂的peg化肝素纳米胶束及其制备方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110954A (zh) * | 2013-01-31 | 2013-05-22 | 北京大学 | 胆固醇修饰的生物可降解聚阳离子载体及制备方法和用途 |
| CN105748439A (zh) * | 2016-01-29 | 2016-07-13 | 四川大学 | 一种基于短链烷烃修饰的树状分子的pH响应型纳米递药系统及其制备方法和应用 |
| CN106491561A (zh) * | 2016-11-10 | 2017-03-15 | 苏州大学 | 基于pamam的联合携载阿霉素及耐药逆转剂的纳米递送系统的制备方法 |
-
2017
- 2017-05-16 CN CN201710343616.2A patent/CN107281161B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110954A (zh) * | 2013-01-31 | 2013-05-22 | 北京大学 | 胆固醇修饰的生物可降解聚阳离子载体及制备方法和用途 |
| CN105748439A (zh) * | 2016-01-29 | 2016-07-13 | 四川大学 | 一种基于短链烷烃修饰的树状分子的pH响应型纳米递药系统及其制备方法和应用 |
| CN106491561A (zh) * | 2016-11-10 | 2017-03-15 | 苏州大学 | 基于pamam的联合携载阿霉素及耐药逆转剂的纳米递送系统的制备方法 |
Non-Patent Citations (1)
| Title |
|---|
| KULKARNI P.，ET AL.: ""Hypoxia-Responsive Polymersomes for Drug Delivery to Hypoxic Pancreatic Cancer Cells"", 《BIOMACROMOLECULES》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111773184A (zh) * | 2020-07-14 | 2020-10-16 | 山东大学 | 一种乏氧响应脂质体及在制备抗肿瘤药物中的应用 |
| CN111773184B (zh) * | 2020-07-14 | 2021-09-28 | 山东大学 | 一种乏氧响应脂质体及在制备抗肿瘤药物中的应用 |
| CN111743861A (zh) * | 2020-07-15 | 2020-10-09 | 南通大学 | 靶向三阴性乳腺癌的低氧响应手性药物胶束及其制备方法 |
| CN111743861B (zh) * | 2020-07-15 | 2022-01-28 | 南通大学 | 靶向三阴性乳腺癌的低氧响应手性药物胶束及其制备方法 |
| CN111821419A (zh) * | 2020-07-31 | 2020-10-27 | 浙江大学 | 一种自组装多肽纳米载体及其制备方法和应用 |
| CN112402625A (zh) * | 2020-11-19 | 2021-02-26 | 健进制药有限公司 | 负载光敏剂的peg化肝素纳米胶束及其制备方法 |
| CN112402625B (zh) * | 2020-11-19 | 2022-12-09 | 健进制药有限公司 | 负载光敏剂的peg化肝素纳米胶束及其制备方法 |
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