CN112972696B - 荧光多肽掺杂金属有机框架纳米材料及制备方法和应用 - Google Patents
荧光多肽掺杂金属有机框架纳米材料及制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种荧光多肽掺杂金属有机框架纳米材料及制备方法和应用。将冷冻的5‑FAM多肽溶液室温下解冻并溶于蒸馏水中得到溶液一，将Zn(NO3)2·6H2O溶于蒸馏水中，置于磁力搅拌器上进行搅拌得到溶液二；将溶液一缓慢加入溶液二中得到混合溶液；室温搅拌0.5‑1h，将2‑甲基咪唑溶液迅速加入到所述混合溶液中；所有操作均在室温下进行；锌离子与2‑甲基咪唑的摩尔比为1:30‑1:80；静置后离心；移除上层清液，得到的颗粒用水冲洗将沉积物在40‑60℃减压干燥12‑15h，得到荧光多肽掺杂金属有机框架纳米材料5‑FP/ZIF‑8。本研究以简单高效的策略将药物装载和检测环节整合到单一平台，为生物医学领域先进药物输送系统的建设开辟了一条新途径。
Description
技术领域
本发明涉及荧光标签修饰的多肽掺杂金属有机框架的制备方法，属于纳米功能材料领域，尤其涉及的是一种荧光多肽掺杂金属有机框架纳米材料及制备方法和应用。
背景技术
ZIF-8是由锌离子与2-甲基咪唑配位组装而成的多孔结晶材料。它具有比表面积大、气孔率高、合成方便、尺寸可控等优点，在功能材料的封装和输送方面具有突出优势。ZIF-8是研究最多的一个MOF材料方钠石(SOD)拓扑结构,表现出一个三维结构形成的锌离子和甲基咪唑链接器。使其具有直径11.6°A的大孔保持架，具有较高的热稳定性，耐水化学性能显著。
目前的大多数抗肿瘤药物存在选择性低、毒副作用强、易产生耐药性等缺点，迫切需要研究和开发新的抗肿瘤药物。人们通过实验发现许多实体瘤的细胞外pH值比正常组织要低。基于这样的机理，设计具有pH响应的肿瘤药物包覆材料，使之能区分正常组织和肿瘤。
目前大多数药物存在选择性低、毒副作用强、易产生耐药等缺点，极大地限制了其临床应用。因此，为了克服这些固有的局限性，进一步满足人们对有效治疗日益增长的需求，设计一种可控的给药系统用于癌症治疗具有重要意义。因此，我们需要一种有效的载药和给药手段来减少对体内正常细胞的损伤。脂质、聚合物纳米粒子、金属团簇和碳结构在过去已经被测试为药物存储材料。然而，这些材料有几个局限性，如载药量低(<5wt％)，药物释放快，毒性大。
发明内容
本发明针对现有技术的不足提供一种荧光多肽掺杂金属有机框架纳米材料及制备方法和应用。通过在ZIF-8上添加荧光标记的组氨酸多肽，我们得到了荧光标记的多肽掺杂的ZIF-8(称为5-FP/ZIF-8)。体外研究表明，5-FP/ZIF-8具有很好的智能pH响应性，组氨酸多肽的添加增强了细胞的通透性。同时，荧光标签在药物的载入和释放时可以产生荧光反应。本研究以简单高效的策略将药物装载和检测环节整合到单一平台，为生物医学领域先进药物输送系统的建设开辟了一条新途径。
本发明的技术方案为：
一种荧光多肽掺杂金属有机框架纳米材料的制备方法，包括以下步骤：
(1)制备荧光标签修饰的多肽掺杂的ZIF-8：将冷冻的5-FAM多肽溶液室温下解冻并溶于蒸馏水中得到溶液一，另外将Zn(NO3)2·6H2O溶于蒸馏水中，置于磁力搅拌器上进行搅拌得到溶液二；将溶液一缓慢加入溶液二中得到混合溶液；室温搅拌0.5-1h，将2-甲基咪唑溶液迅速加入到所述混合溶液中；所有操作均在室温下进行；锌离子与2-甲基咪唑的摩尔比为1:30-1:80；
(2)静置24-36h,6000-8000rpm离心；移除上层清液，得到的颗粒用水冲洗将沉积物在40-60℃减压干燥12-15h，得到荧光多肽掺杂金属有机框架纳米材料5-FP/ZIF-8。
所述的制备方法，步骤(1)中：5-FAM肽与锌离子的摩尔比为1：250-500。
所述的制备方法，步骤(1)所加入的荧光标签5-FAM替换为6-FAM、FITC或cy5。
所述的制备方法，步骤(1)，加入500uL 0.25mol/L的Zn(NO3)2·6H2O溶液，5mL2mol/L的2-甲基咪唑溶液。
所述的制备方法，步骤(2)所述的将沉淀溶液离心中，为中低速离心，离心机转速为6000-8000转/分，时间为20-30min。
所述的制备方法，步骤(2)中沉淀洗涤三次，直至呈微碱性，为充分洗涤每次超声5-10min。
所述的制备方法，步骤(2)中减压干燥时温度不超过60℃，直至彻底干燥。
根据任一所述的制备方法获得的荧光多肽掺杂金属有机框架纳米材料。
所述的荧光多肽掺杂金属有机框架纳米材料在制备抗癌药物中的应用。
所述的应用，所述抗癌药物为阿霉素、柔红霉素或环磷酰胺。
本发明的有益效果是：5-FP/ZIF-8纳米复合材料可以作为一种高效的pH响应型药物传递系统，在pH为6.3时能够缓慢释放所载药物。5-FAM荧光标记可有效检测药物的装载和释放，5-FAM的荧光强烈依赖于药品载入的量，随药物载入量的增加荧光减弱。当药物释放后荧光恢复。由于组氨酸多肽的存在，细胞膜的穿透和破裂增加了其通透性，这更有利于药物特异性识别和进入癌细胞，降低活性，加速癌细胞的死亡。可为开发一种用于癌症治疗的智能pH敏感给药系统提供新的机会。
附图说明
图1为带有荧光标签修饰的多肽掺杂金属有机框架的制备方法与应用的流程图。
图2为5-FAM多肽掺杂ZIF-8的SEM图像。
图3为TEM图像。
图4为5-FAM多肽掺杂ZIF-8的荧光激发发射光谱。
图5为5-FAM多肽掺杂ZIF-8载入阿霉素的荧光淬灭光谱。
图6为5-FAM多肽掺杂ZIF-8浸泡于pH为6.3的PBS溶液的荧光恢复光谱。
图7为5-FAM多肽掺杂ZIF-8浸泡于pH为6.8的PBS溶液的荧光恢复光谱。
图8为负载阿霉素的5-FAM多肽掺杂ZIF-8浸泡于pH为6.3的PBS溶液的透射图。
图9为细胞实验的共聚焦显微镜图。
具体实施方式
以下结合具体实施例，对本发明进行详细说明。
实施例1
制备荧光标签修饰的多肽掺杂的ZIF-8：将冷冻的5-FAM多肽溶液室温下解冻，另外将其溶于蒸馏水中，置于磁力搅拌器上进行搅拌。将Zn(NO3)2·6H2O配成0.25mol/L的水溶液。另外，将2-甲基咪唑配成1mol/L的水溶液。取5uM的5-FAM多肽100uL加入到500uLZn(NO3)2·6H2O溶液中搅拌10min，将5mL甲基咪唑溶液迅速加入到上述溶液中，室温搅拌0.5h，所有操作均在室温下进行，锌离子与甲基咪唑的摩尔比为1:40。混合各组分溶液后，混合物立即变得混浊。静置24-36h,6000-8000rpm离心10min。移除上层清液。得到的颗粒用水冲洗三次；最后，将沉积物在40-60℃减压干燥12-15h。
称取2g 5-FAM标记的多肽掺杂的ZIF-8固体并加入2ml水，超声5-10min，然后加入0～100μM不同浓度的阿霉素标准溶液，5分钟后离心，离心机转速为6000转/分，移走上清液，真空干燥，温度不超过40℃，直至彻底干燥。
图2为5-FAM标记的多肽掺杂的ZIF-8(5-FP/ZIF-8)固体的扫描图，图3为5-FAM标记的多肽掺杂的ZIF-8固体的透射图，我们可以发现5-FP/ZIF-8为200纳米左右的固体颗粒，与ZIF-8固体相似，没有明显的形态变化，说明ZIF-8中的肽包覆并不影响ZIF-8的基本结构。图4为5-FAM标记的多肽掺杂的ZIF-8固体制备过程中上清液与形成的固体颗粒的荧光激发和发射谱图。我们发现离心后的5-FP/ZIF-8上清液只有微弱的荧光，下部沉淀的固体颗粒与5-FAM荧光标签的荧光发射峰相同，说明5-FAM标记的多肽成功掺杂进ZIF-8固体中。图5为不同浓度阿霉素溶液浸泡下的荧光发射谱图。将ZIF-8的固体颗粒分散在水溶液中，并浸入不同浓度的阿霉素中进行进一步的发光研究。ZIF-8的发光强度强烈地依赖于阿霉素浓度。发光强度随阿霉素浓度的增加而成比例下降，说明5-FAM标记的多肽掺杂的ZIF-8固体可用来负载及传感药物的负载情况。
实施例2
制备荧光标签修饰的多肽掺杂的ZIF-8：将冷冻的5-FAM多肽溶液室温下解冻，另外将溶于蒸馏水中，置于磁力搅拌器上进行搅拌。将Zn(NO3)2·6H2O配成0.25mol/L的水溶液。另外，将2-甲基咪唑配成2mol/L的水溶液。取5uM的5-FAM多肽100uL加入到500uLZn(NO3)2·6H2O溶液中搅拌10min，将5mL甲基咪唑溶液迅速加入到上述溶液中。室温搅拌0.5h。所有操作均在室温下进行。锌离子与甲基咪唑的摩尔比为1:80。混合各组分溶液后，混合物立即变得混浊。静置24-36h,6000-8000rpm离心10min。移除上层清液。得到的颗粒用水冲洗三次；；最后，将沉积物在40-60℃减压干燥12-15h。
称取2g 5-FAM多肽ZIF-8固体并加入2ml水。超声5-10min，然后加入不同浓度的阿霉素标准溶液(0～100μM)。室温孵育30s后，加入石英荧光试管。在494nm激发下，在500～700nm范围内测量荧光光谱。标签的荧光强烈依赖于药品载入的量，随药物载入量的增加荧光减弱。5分钟后离心。称取负载药物的ZIF-8固体分散于pH为6.3和6.8磷酸盐缓冲液(PBS)中。每隔30分钟使用荧光光谱在pH为6.3的PBS缓冲液中每隔30分钟用494nm激发，在500～700nm范围内测量荧光光谱。
图6和图7分别为负载了阿霉素的5-FP/ZIF-8固体在pH为6.3和6.8的PBS缓冲液中浸泡不同时间的荧光发射图，图8为pH为6.3透射图。随着固体颗粒逐渐破裂阿霉素的释放荧光逐渐增强，pH为6.3时0-12小时内荧光强度明显强于pH为6.8。说明5-FP/ZIF-8固体具有pH敏感性，在pH为6.3时可以有效释放药物，具有pH敏感性且可以有效传感药物的释放。
实施例3
制备荧光标签修饰的多肽掺杂的ZIF-8：将冷冻的5-FAM多肽溶液室温下解冻，另外将溶于蒸馏水中，置于磁力搅拌器上进行搅拌。将Zn(NO3)2·6H2O配成0.25mol/L的水溶液。另外，将2-甲基咪唑配成2mol/L的水溶液。取5uM的5-FAM多肽100uL加入到500uLZn(NO3)2·6H2O溶液中搅拌10min，将5mL甲基咪唑溶液迅速加入到上述溶液中。室温搅拌0.5h。所有操作均在室温下进行。锌离子与甲基咪唑的摩尔比为1:60。混合各组分溶液后，混合物立即变得混浊。静置24-36h,6000-8000rpm离心10min。移除上层清液。得到的颗粒用水冲洗三次；最后，将沉积物在40-60℃减压干燥12-15h。称取5-FAM标记的多肽掺杂的ZIF-8固体并加入2ml水。超声5-10min，然后加入不同浓度的阿霉素标准溶液(0～100μM)。5分钟后离心。移走上清液。
分别用掺杂5-FAM标记的多肽ZIF-8颗粒和只掺杂5-FAM标签的ZIF-8颗粒分别培养的Hep G2细胞。取对数生长期的Hep G2细胞，进行细胞计数，调整细胞浓度，分别称取0.1mg样品，溶解于1mL完全培养基中，超声15min，放置于30mm培养皿中紫外照射30min灭菌，得到浓度为0.1mg/mL的工作液，4℃保存备用。按照上述分组进行处理。5％CO 2，37℃恒温培养箱中分别培养2h；1mL/皿PBS清洗3次。按照1mL/皿加入PBS，上机拍摄。
图9为细胞实验的共聚焦显微镜的图像，图(b)为用掺杂5-FAM标记的多肽ZIF-8颗粒培养的细胞图像，图(c)为用只掺杂5-FAM标签的ZIF-8颗粒培养的细胞图像。我们发现，图(b)的荧光明显多于图(c),说明多肽的加入增加了细胞的通透性。
应当理解的是，对本领域普通技术人员来说，可以根据上述说明加以改进或变换，而所有这些改进和变换都应属于本发明所附权利要求的保护范围。
Claims (8)
1.一种荧光多肽掺杂金属有机框架纳米材料的制备方法，其特征在于，包括以下步骤：
（1）制备荧光标签修饰的多肽掺杂的ZIF-8：将冷冻的5-FAM修饰的组氨酸多肽溶液室温下解冻并溶于蒸馏水中得到溶液一，将Zn(NO3)2溶于蒸馏水中，置于磁力搅拌器上进行搅拌得到溶液二；将溶液一缓慢加入溶液二中得到混合溶液；室温搅拌0.5-1h，将2-甲基咪唑溶液迅速加入到所述混合溶液中；所有操作均在室温下进行；锌离子与2-甲基咪唑的摩尔比为1:30-1:80；所述组氨酸多肽能够增强细胞的通透性；
（2）静置后离心；移除上层清液，得到的颗粒用水冲洗将沉积物在40-60℃减压干燥，得到荧光多肽掺杂金属有机框架纳米材料5-FP/ZIF-8；
（3）在5-FP/ZIF-8中装载药物阿霉素。
2.根据权利要求1所述的制备方法，其特征在于，步骤（1）中：5-FAM肽与锌离子的摩尔比为1：250-500。
3.根据权利要求1所述的制备方法，其特征在于，步骤（1）所述加入500uL 0.25 mol/L的Zn(NO3)2·6H2O溶液，5mL 2mol/L的2-甲基咪唑溶液。
4.根据权利要求1所述的制备方法，其特征在于，步骤（2）所述的将沉淀溶液离心中，为中低速离心，离心机转速为6000-8000转/分，时间为20-30min。
5.根据权利要求1所述的制备方法，其特征在于，步骤（2）中沉淀洗涤三次，直至呈微碱性，为充分洗涤，每次超声5-10min。
6.根据权利要求1所述的制备方法，其特征在于，步骤（2）中减压干燥时温度不超过60℃，直至彻底干燥。
7.根据权利要求1-6任一所述的制备方法获得的荧光多肽掺杂金属有机框架纳米材料。
8.根据权利要求7所述的荧光多肽掺杂金属有机框架纳米材料在制备抗癌药物中的应用。
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Non-Patent Citations (3)
| Title |
|---|
| Jiaming Xuan et al..Peptide-Mediated Synthesis of Zeolitic Imidazolate Framework-8 with Controllable Morphology and Size.《Langmuir》.2020,第36卷(第46期), * |
| Pengcheng Zhang et al..Cellular Uptake and Cytotoxicity of Drug−Peptide Conjugates.《Bioconjugate Chemistry》.2013,第24卷(第4期), * |
| Yixuan Lin et al..Ligand-Modified Erythrocyte Membrane-Cloaked Metal−Organic Framework Nanoparticles for Targeted Antitumor Therapy.《Molecular Pharmaceutics》.2020,第17卷(第9期), * |
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