CN111687428A - 两亲性聚合物介导金纳米粒子可控组装体及其制备与应用 - Google Patents
两亲性聚合物介导金纳米粒子可控组装体及其制备与应用 Download PDFInfo
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- CN111687428A CN111687428A CN202010402495.6A CN202010402495A CN111687428A CN 111687428 A CN111687428 A CN 111687428A CN 202010402495 A CN202010402495 A CN 202010402495A CN 111687428 A CN111687428 A CN 111687428A
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- amphiphilic polymer
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- mediated gold
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Abstract
本发明属于光学纳米材料的技术领域,公开了两亲性聚合物介导金纳米粒子可控组装体及其制备与应用。所述方法:1)两亲性聚合物和烷基硫醇在水中形成胶束;加入四氯金酸,混合均匀,获得Au(I)硫醇复合物;所述两亲性聚合物为泊洛沙姆F127;2)调节Au(I)硫醇复合物的pH至碱性,加入还原剂进行还原反应,后续处理,获得两亲性聚合物介导的金纳米组装体。本发明的方法简单,成本低,易于工业化生产;制备金纳米组装体生物相容性好,毒性低,可长时间在体内血液循环,并具有近红外发光特点,稳定性好,激发波长范围广。本发明的金纳米组装体用于医药领域、生物传感器、生物传感检测领域、荧光成像领域以及肿瘤成像的成像剂。
Description
技术领域
本发明属于功能光学纳米材料领域,具体涉及两亲性聚合物介导金纳米组装体及其制备与应用。
背景技术
两亲性嵌段聚合物因其特殊尺寸和形貌,常被用来作为临床疏水药物(阿霉素和紫杉醇等)的载体。负载在胶束核内的疏水药物因外层亲水基团的保护,可避免被肝和脾捕获之后快速排出体外,达到长时间体内长循环的作用,从而实现高肿瘤靶向的目标。这类具有核壳结构载体的形貌有球形、长条形和纤维状,而形貌对于小分子药物的运送产生极大的影响。相对于小分子药物,被嵌段聚合物负载的无机纳米材料在活体内的相关研究如生物分布、细胞内化和肿瘤靶向等仍处于初级阶段。
发光金纳米粒子具有超小的尺寸、表面易修饰及良好的生物相容性等优点,在传感检测、生物成像及疾病诊断等领域极具应用潜力。特别是,动物的组织和细胞对近红外区域的光源有较少的吸收,使得在近红外区发光的超小金纳米粒子在生物医药和临床治疗领域具备巨大的应用价值。由于发光金纳米粒子的尺寸小于6nm,可通过肾脏迅速排出体外,可以防止其因长时间体内滞留造成机体伤害;然而,血液循环时间短会造成纳米材料的靶向效率低。
近年来,金纳米粒子作为基元构建的新型组装体复合材料,因所展现的独特光学性质,备受科学工作者的广泛关注。不同形貌和功能的金纳米粒子组装体已被成功制得。相比于单个金纳米粒子,金纳米粒子组装体在近红外波段发光更强,使其在生物、医药等领域有了更为重要的应用。另外,金纳米粒子与嵌段聚合物组装成的纳米复合结构不仅可以集成各个结构基元的功能,而且可以具备新的性质与功能,为构建多功能的纳米复合材料提供了有效可行的途径。然而,要获得一种发光波长可调、形貌和装载量可控的发光金纳米组装体是现阶段的一大挑战。另外,组装体的有关性质(如形貌和装载量)与它在活体内的递送关系仍然处于未知状态。因此,发展一种光学、形貌和装载量可控的纳米组装结构的有效制备方法对生物、医药领域具有重要的意义。
发明内容
为了克服现有技术的缺点与不足,本发明的首要目的在于提供一种两亲性聚合物介导金纳米组装体的制备方法。
本发明的另一目的在于提供由上述制备方法得到的两亲性聚合物介导的金纳米组装体。所述的两亲性聚合物介导的金纳米组装体具有近红外发光性质,便于可视化研究,且自组装体具有发光波长可调,组装体形貌和金纳米粒子装载量可调,稳定性好、生物毒性低。
本发明的再一目的在于提供上述两亲性聚合物介导的金纳米组装体的应用。所述两亲性聚合物介导的金纳米组装体用于载药领域,用作药物的载体;还可用于生物传感器和以非疾病治疗和非疾病诊断为目的荧光成像领域以及肿瘤成像的成像剂。本发明的两亲性聚合物介导的金纳米在活体内可实现长时间血液循环,可将组装体应用于载药,实现诊疗一体化;所述纳米材料由于近红外发光和高肿瘤靶向特点,可原位观测肿瘤在活体内的分布情况(例如通过活体成像仪等)。
本发明的目的通过下述技术方案实现:
一种两亲性聚合物介导的金纳米组装体的制备方法,包括如下步骤:
1)两亲性聚合物和烷基硫醇在水中形成胶束;加入四氯金酸,混合均匀,获得Au(I)硫醇复合物;所述两亲性聚合物为泊洛沙姆F127;所述烷基硫醇为己硫醇、十二烷基硫醇或十八烷基硫醇中一种以上;
2)调节Au(I)硫醇复合物的pH至碱性,加入还原剂进行还原反应,后续处理,获得两亲性聚合物介导的金纳米组装体;所述还原剂为四羟甲基氯化磷(THPC)、二甲基胺硼烷(DMAB)、硼氢化钠(NaBH4)中一种以上。
所述pH为8~13。
所述氯金酸与配体烷基硫醇的摩尔比为1:(1~5),优选为1:(2~4);氯金酸与还原剂的摩尔比为(25~220):1。
还原剂为四羟甲基氯化磷(THPC)时,氯金酸与还原剂的摩尔比为(130~160):1
还原剂为二甲基胺硼烷(DMAB)时,氯金酸与还原剂的摩尔比为(180~220):1;
还原剂为硼氢化钠(NaBH4)时,氯金酸与还原剂的摩尔比为(25~50):1。
所述还原反应的时间为3~24h。
步骤1)中所述两亲性聚合物与水的质量比为1:(40~60)。
所述烷基硫醇与两亲性聚合物的用量关系为(0.3~1.5)mmol:1g。即每1g的两亲性聚合物加入(0.3~1.5)mmol的烷基硫醇。
所述后续处理是指离心,透析纯化。所述透析是指在水中透析,透析的时间为1~5天,所述透析的透析袋的截留分子量为1~50kDa。
透析后进行超滤浓缩,超滤浓缩使用超滤管的膜孔径为3~50kDa。
氯金酸,还原剂,烷基硫醇都可以溶液的形式加入。
两亲性聚合物和烷基硫醇在水中形成胶束是指将两亲性聚合物和烷基硫醇与水混合,搅拌形成胶束;或者将两亲性聚合物加入水中,然后加入烷基硫醇。
所述两亲性聚合物介导的金纳米组装体在室温下保存。
本发明的两亲性聚合物介导的金纳米组装体的形貌为组装体结构,所述的两亲性聚合物和金纳米粒子自组装形成长条状和球状结构,金纳米粒子以颗粒状被包裹于所述的纳米组装体结构中,所述的金纳米单颗粒子粒径为1~3nm,所述的金纳米组装体的长度为10~50nm,宽度为10~25nm。
所述两亲性聚合物介导的金纳米组装体,通过上述制备方法制得。
所述的金纳米组装体的长度为10~50nm,宽度为10~25nm;组装体中金纳米单颗粒子粒径为1~3nm。
所述两亲性聚合物介导的金纳米组装体用于医药领域,特别是载药领域,用作药物的载体;所述药物优选为抗肿瘤的药物。所述两亲性聚合物介导的金纳米组装体用于制备高肿瘤靶向药物制剂。
所述两亲性聚合物介导的金纳米组装体还可用于生物传感器、生物传感检测领域,以非疾病治疗和非疾病诊断为目的。
所述两亲性聚合物介导的金纳米组装体用于荧光成像领域以及肿瘤成像的成像剂,以非疾病治疗和非疾病诊断为目的。所述肿瘤成像的成像剂是指高肿瘤靶向的成像剂。
所述的两亲性聚合物介导的金纳米组装体在药代动力学、生物分布和活体成像方面的应用。
本发明以两亲性聚合物为模板合成金纳米发光材料,在金纳米粒子具有生物相容性好,毒性小,光学性能优良的基础上,通过本发明成功实现了嵌段聚合物对金纳米粒子负载量的调控。本发明用不同的简单易得而且绿色无毒的还原剂即可得到不同激发波长和发射波长的两亲性聚合物介导的金纳米组装体。
本发明合成的两亲性聚合物介导的金纳米组装体生物相容性好,毒性低,在药代动力学方面,具有良好的效果;而此两亲性聚合物介导的金纳米组装体在室温、DMEM培养基等条件下荧光稳定性好,量子效率高,用超敏多功能成像仪可以明显的观察到材料在体内分布的情况,故此两亲性聚合物介导的金纳米组装体在载药、肿瘤成像等领域具有较大的应用前景。
本发明实现了嵌段聚合物载体的可视化研究,用于生物传感及荧光成像等领域研究,具有灵敏度高、重复性好和操作简便等特点;同时,由于引入了具有超小的尺寸和独特的光学性质纳米金材料,也可通过电感耦合等离子体质谱仪来定量,在肿瘤靶向及肿瘤成像等领域具有良好的应用前景。
本发明的两亲性聚合物介导的金纳米组装体发射波长范围为400~900nm,激发波长范围为300~650nm。
本发明相对于现有技术具有如下的优点及效果:
(1)本发明的方法简单,成本低,易于工业化生产。
(2)本发明合成的两亲性聚合物介导的金纳米组装体生物相容性好,毒性低,可长时间在体内血液循环,可用于载药。
(3)本发明合成的两亲性聚合物介导的金纳米组装体具有近红外发光特点,稳定性好,激发波长范围广,可以很好的应用于活体成像,具有可视化示踪功能(例如,通过超敏多功能成像仪观察),用于研究肿瘤分布,操作简单。
附图说明
图1为本发明的两亲性聚合物介导的金纳米组装体的制备流程示意图;
图2为实施例1中合成的产物的荧光发射光谱以及紫外吸收光谱图;
图3为实施例1中合成的产物的红外光谱图;
图4为实施例1中合成的两亲性聚合物介导的金纳米组装体的X射线光电子能谱图;
图5为实施例1中合成的两亲性聚合物介导的金纳米组装体的透射电子显微镜图;
图6为实施例1中合成的两亲性聚合物介导的金纳米组装体的单颗金纳米粒子的粒径统计图;
图7为实施例1中合成的两亲性聚合物介导的金纳米组装体的扫描电子显微镜图;
图8为实施例1中合成的两亲性聚合物介导的金纳米组装体的胶束的尺寸统计图;
图9为实施例1中合成的两亲性聚合物介导的金纳米组装体在室温条件下荧光稳定性图;
图10为实施例1中合成的两亲性聚合物介导的金纳米组装体在DMEM培养基荧光稳定性图;
图11为实施例2中两亲性聚合物介导的金纳米组装体和牛血清蛋白孵化后的水合粒径图;
图12为实施例3中两亲性聚合物介导的金纳米组装体在不同浓度下的毒性实验结果分析图;
图13为实施例3中两亲性聚合物介导的金纳米组装体在不同时间内被癌细胞摄取的量的柱状图;
图14为实施例4中为两亲性聚合物介导的金纳米组装体在小鼠体内的药代动力学趋势图;
图15为实施例4中两亲性聚合物介导的金纳米组装体在小鼠体内的药代动力学结果分析图;
图16为实施例5中两亲性聚合物介导的金纳米组装体在小鼠体内的生物分布图;
图17为实施例6中两亲性聚合物介导的金纳米组装体在小鼠体内的活体肿瘤成像图;
图18为对比例1中合成的产物的荧光发射光谱以及紫外吸收光谱图;左图为十二烷基硫醇为配体时合成的产物的光谱图,右图为十八烷基硫醇为配体时合成的产物的光谱图;
图19为对比例2中合成的产物的荧光发射光谱以及紫外吸收光谱图;
图20为对比例3中四氯金酸与己硫醇的摩尔比为1:1.5时合成的产物的透射电子显微镜图
图21为对比例3中四氯金酸与己硫醇的摩尔比为1:5时合成的产物的透射电子显微镜图。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述,但本发明的实施方式不限于此。
在以下具体实施例中,所涉及到的MDA-MB-231细胞可购自ATCC,小鼠可购于广东省实验动物中心,DMEM培养基(gibico,货号:LOT8118192),胎牛血清(FBS,LONSA SCIENCESRL,CAS号:S711-001S)。观察两亲性聚合物介导的金纳米组装体发光性质,形貌及其在小鼠药代动力学与肿瘤成像分析等方面的仪器主要包含日本日立公司扫描电子显微镜(SU8220)、日本电子株式会社JEM-2100F透射电子显微镜、美国PerkinElmer荧光/磷光/发光光度计(LS-55)、超敏多功能成像仪系统以及德国Thermo Scientific电感耦合等离子体质谱仪(iCAP RQ)等。
实施例1
两亲性聚合物介导的金纳米组装体材料通过以下方法制备:
在室温条件下,将18.75μL己硫醇(8M)(溶剂为水)、0.2g F127和10mL水加入25mL三口烧瓶中,充分搅拌(搅拌0.5h)使其形成胶束,而具有疏水性的配体和F127的疏水嵌段通过疏水相互作用行进自主装;随后加入1mL四氯金酸(50mM,溶剂为水),室温剧烈搅拌(搅拌的转速为1000rpm)至无色形成金一价硫醇复合物后,先使用0.1M的氢氧化钠将溶液的pH调至11,加入还原剂反应(加入6.8μL四羟甲基氯化磷(50mM)室温下磁力搅拌反应8h形成发光位置在520nm的两亲性聚合物介导的金纳米组装体(AuNAs-520);或者加入10μL二甲基胺硼烷(25mM)室温下磁力搅拌反应12h形成发光位置在610nm的两亲性聚合物介导的金纳米组装体(AuNAs-610);或者加入30μL硼氢化钠(50mM)在室温下磁力搅拌分别反应5h形成发光位置在810nm的两亲性聚合物介导的金纳米组装体(AuNAs-810)),磁力搅拌转速为1000rpm,溶液由无色分别到绿色、浅黄色、棕黄色,停止反应,先用21000g高速离心机离心15min,除去反应生成的大颗粒,然后用透析袋在pH为中性的水中透析2天去除未反应的配体、游离的F127和多余的小分子和离子,得到发光分别为520nm、610nm和810nm的目标产物(即AuNAs-520,AuNAs-610,AuNAs-810),在室温下储存备用。
结构表征及荧光稳定性测试:
图1为两亲性聚合物介导的金纳米组装体的制备流程示意图。图1依次展示了本发明的组装体本身的结构,自组装原因,所述嵌段聚合物自组装前形成的形貌和加入不同还原剂之后的组装形貌。
图2为本实施例所制得的两亲性聚合物介导的金纳米组装体的紫外吸收光谱和荧光发射光谱(图中520nm、610nm和810nm分别对应AuNAs-520,AuNAs-610,AuNAs-810)。从荧光发射光谱图可知,三种产物分别在520nm、610nm和810nm处有最大发射峰。
图3为本实施例合成的两亲性聚合物介导的金纳米组装体的红外光谱图。从图3中可知,己硫醇(C6-SH)的特征峰有在2570cm-1位置的S-H的对称伸缩振动,泊洛沙姆F127的特征峰有在2880cm-1位置的C-H的对称伸缩振动和在1105cm-1的C-O-C的对称伸缩振动。合成完成之后的AuNAs-520、AuNAs-610和AuNAs-810消失的己硫醇特征峰表明己硫醇与金粒子成功以金硫键的方式修饰在AuNPs表面;均具有泊洛沙姆F127的特征峰则表明泊洛沙姆F127成功与金纳米粒子成功完成了自组装。
图4为本实施例合成的两亲性聚合物介导的金纳米组装体的X射线光电子能谱图,Au(0)含量比Au(I)要少,AuNAs-520、AuNAs-610和AuNAs-810对应的Au(I)所占的比例分别为40.81%,36.53%和34.35%,这个结果表明随着AuNPs(金纳米粒子)表面连接的配体增加Au(I)含量也在增加从而导致了发射光的蓝移。
图5为本实施例合成的两亲性聚合物介导的金纳米组装体的透射电子显微镜图。如图5所示,所述的两亲性聚合物介导的金纳米组装体,其组装形貌为分别长条形(AuNAs-520)、长条形和球形(AuNAs-610)、球形(AuNAs-810),而不同形貌上的每个黑色颗粒表示为在组装体上生长的金纳米颗粒。
通过粒径分析软件(如Nano Measurer 1.2.5)对合成的两亲性聚合物介导的金纳米组装体的单颗金纳米粒子进行统计,结果如图6所示,其粒径都约为1.7nm。
图7为本实施例合成的两亲性聚合物介导的金纳米组装体的扫描电子显微镜图。从图7中,可清晰观察到它们的整体形貌,其组装形貌为分别长条形(AuNAs-520)、长条形和球形(AuNAs-610)、球形(AuNAs-810)。
通过粒径分析软件(如Nano Measurer 1.2.5)对合成的两亲性聚合物介导的金纳米组装体的胶束进行统计,见图8。图8为实施例1中合成的两亲性聚合物介导的金纳米组装体的胶束的尺寸统计图。从数据中可知AuNAs-520的长为33.5±10.0nm,宽为16.4±3.4nm、AuNAs-610的长为25.1±5.5nm,宽为16.6±3.8nm、AuNAs-810的粒径为21.9±5.2nm。
考察本实施例制得的两亲性聚合物介导的金纳米组装体材料的荧光在不同溶剂中的稳定性:
(1)在室温条件下的荧光稳定性研究
取100μL所述的合成的两亲性聚合物介导的金纳米组装体,分别稀释至1mL,用荧光发光光度计检测在不同时间点的荧光。
(2)在DPBS条件下的荧光稳定性研究
取100μL所述的合成的两亲性聚合物介导的金纳米组装体,分别加入含10%FBS(v/v)的DPBS溶液稀释至1mL,用荧光发光光度计检测在不同时间点的荧光。
图9为实施例1中合成的两亲性聚合物介导的金纳米组装体在室温条件下荧光稳定性图;图10为实施例1中合成的两亲性聚合物介导的金纳米组装体在DMEM培养基荧光稳定性。
图9可知,合成的两亲性聚合物介导的金纳米组装体在室温条件下荧光稳定性好。图10可知,合成的两亲性聚合物介导的金纳米组装体在DPBS溶液条件下荧光稳定性好。
实施例2
取100μL所述的合成的两亲性聚合物介导的金纳米组装体(溶剂为水),分别加入100μL的FBS 10%(v/v)并稀释至1mL,对照组直接将材料稀释(采用水稀释)至1mL,在37℃的孵化箱中孵化30分钟,之后用粒度仪(Malvern Mastersizer)测它们的水合粒径。从而辨别它们的FBS的吸附能力。
图11为实施例2中两亲性聚合物介导的金纳米组装体和牛血清蛋白孵化后的水合粒径图。图11为两亲性聚合物介导的金纳米组装体本身的水合粒径,还有和FBS在37℃条件下孵化30min后的水合粒径,图中AuNAs-520的水合粒径在与FBS培养之后从30.8±5.5nm增长到50.0±10.9nm,AuNAs-610的水合粒径在与FBS培养之后从29.6±3.4nm增长到30.8±6.0nm,AuNAs-810的水合粒径在与FBS培养之后从20.6±3.5nm增长到21.7±4.1nm。从以上结果可知两亲性聚合物介导的金纳米组装体本身具有的抗蛋白吸附能力顺序为:AuNAs-520<AuNAs-610<AuNAs-810。
实施例3
本发明制备两亲性聚合物介导的金纳米组装体(实施例1制备)用于探测MDA-MB-231乳腺癌细胞对其的毒性和摄取能力:将10μL不同浓度的实施例1制备的纳米材料和90μLDMEM培养基混合,和细胞培养24h,通过MTT检测试剂盒对两亲性聚合物介导的金纳米组装体进行在不同浓度下的毒性实验研究。
将10μL实施例1制备的纳米材料(5mg/L)和490μL DMEM培养基混合,分别与细胞培养1、3、6、12、24h后,吸走培养液,用DPBS洗涤3次,加入新制的王水溶解细胞里的金纳米组装体,之后用电感耦合等离子体测其中的金元素含量。
图12为两亲性聚合物介导的金纳米组装体在不同浓度下的细胞毒性实验分析结果图。如图12所示材料基本无毒。
图13为MDA-MB-231细胞在不同时间下对两亲性聚合物介导的金纳米组装体的摄取量柱状图,其中,AuNAs-520被摄取能力大于AuNAs-610和AuNAs-810。以上结果表明AuNAs的表面亲疏水性是细胞对长条形AuNAs-520摄取的一个重要因素,因为更疏水的表面会吸附更多的蛋白质,而细胞正好可以识别蛋白质,吸附了蛋白质的材料被细胞识别从而被摄取。容易被细胞摄取的长条形AuNAs-520会因此无法在血液中实现长时间循环。
实施例4
本发明制备两亲性聚合物介导的金纳米组装体用于探测其在小鼠体内的药代动力学:
通过尾静脉往小鼠注入实施例1制备的纳米组装体材料,然后在不同时间点从小鼠的眼眶静脉取出大约20μL的血,置于玻璃瓶中,称量并标记好,用新制王水对样品进行硝解,接着用电感耦合等离子体测出金含量,计算得到各个时间点实施例1制备的纳米材料在血液中的含量。
图14为两亲性聚合物介导的金纳米组装体在小鼠体内的药代动力学趋势图。如图14所示,材料具有长时间循环的优势。
图15为两亲性聚合物介导的金纳米组装体在小鼠体内药代动力学的曲线下面积统计(药代动力学结果分析图),它代表材料的长时间循环能力。
实施例5
本发明制备两亲性聚合物介导的金纳米组装体用于探测其在小鼠体内的生物分布:
为了得到具体的实施例1制备的纳米材料在活体内的分布量,在注入两亲性聚合物介导的金纳米组装体之后不同时间点处死小鼠,取出主要器官(如心、肝、脾、肺、肾)和肿瘤,用新制王水硝解,接着用电感耦合等离子体测出金含量,计算得到实施例1制备的纳米材料在各个器官和肿瘤的分布含量。
图16为两亲性聚合物介导的金纳米组装体在小鼠体内的生物分布图。从图中可知材料具有较高的肿瘤靶向能力(金纳米组装体可以从肝脾等器官转移出来然后通过粪便里排出体外,图16所获得的器官是在注射72h后处死小鼠得到的)。
实施例6
本发明制备两亲性聚合物介导的金纳米组装体用于探测其在小鼠肿瘤成像:
为了得到具体的实施例1制备的在近红外发光的纳米材料肿瘤成像的效果,在往小鼠身上注射材料之前先给带有肿瘤的小鼠成像作为对照组,在对荷瘤小鼠尾静脉注射AuNAs-810后的不同时间对小鼠进行荧光成像。
图17为两亲性聚合物介导的金纳米组装体在小鼠肿瘤成像的效果图。从图中可知材料具有较高的肿瘤靶向能力且可长时间肿瘤靶向。
从图17中可以看到,在扣除背景之后,可看到在注射材料之前,小鼠的身上没有明显的荧光;在注射10min后,材料迅速遍布小鼠全身,小鼠全身开始出现微弱荧光;在注射1h后,小鼠的膀胱出现荧光说明一部分金纳米材料通过肾脏排出体外;注射3h后,也可以看到小部分金纳米粒子通过肾脏排出体外;注射3h后,可以观察到肿瘤部位出现比身体其他地方更亮的荧光;注射24~96h后,可以明显观察到小鼠的肿瘤部位的荧光,说明材料可以实现长时间肿瘤成像。
对比例1(采用其他硫醇)
采用十二烷基硫醇替换实施例1中配体己硫醇,其他条件同实施例1,分别得到AuNAs-550、AuNAs-640和AuNAs-810;
采用十八烷基硫醇分别替换实施例1中配体己硫醇,其他条件同实施例1,分别得到AuNAs-625、AuNAs-650和AuNAs-810。
图18为对比例1中合成的产物的荧光发射光谱以及紫外吸收光谱图;左图为十二烷基硫醇为配体时合成的产物的光谱图,右图为十八烷基硫醇为配体时合成的产物的光谱图。
在使用十二烷基硫醇和十八烷基硫醇作为金纳米粒子的配体时,我们发现它们均无法和己硫醇一样得到峰宽较窄的荧光峰,说明这两个配体合出来的材料混合物较多,不够纯。
对比例2(采用其他还原剂)
采用其他还原剂(硼烷、水合肼、维生素A、柠檬酸钠、甲苯、甲醛)替换实施例1中的还原剂(1.5*10-3mmol),其他条件同实施例1。
图19为对比例2中合成的产物的荧光发射光谱以及紫外吸收光谱图。
对比例3(用量不同)
(1)四氯金酸与己硫醇的摩尔比为1:1.5,其他条件同实施例1,分别得到AuNAs-500、AuNAs-600和AuNAs-800。
对比例3中四氯金酸与己硫醇的摩尔比为1:1.5时合成的产物的透射电子显微镜图如图20所示。
(2)四氯金酸与己硫醇的摩尔比为1:5,其他条件同实施例1,分别得到AuNAs-500、AuNAs-600和AuNAs-800。
对比例3中四氯金酸与己硫醇的摩尔比为1:5时合成的产物的透射电子显微镜图如图21所示。
本发明的两亲性聚合物介导的金纳米组装体在药代动力学方面的应用,具体方法为:通过尾静脉往小鼠体内注入实施例1制备的纳米材料,然后在不同时间点从小鼠的眼眶静脉取血,置于提前准备好的玻璃瓶,称量并标注好,用新制王水硝解样品,并用电感耦合等离子体质谱仪测出样品的金含量,计算得到各个时间点实施例1制备的纳米材料在血液中的含量。
所述两亲性聚合物介导的金纳米组装体使用的浓度为1~20mg/mL,取出的血液体积大约20μL,实验时间0~96h。王水硝解时间为1~24h。所述待测小鼠为balb/c小鼠,而不仅限于balb/c小鼠。
所述的两亲性聚合物介导的金纳米组装体在生物分布方面的应用,具体方法为:通过尾静脉往带有皮下瘤的免疫缺陷裸鼠体内注入实施例1制备的纳米材料,然后在不同时间处死小鼠并取出器官和肿瘤,用新制王水硝解,并用电感耦合等离子体质谱仪测出金含量,计算得到各个时间点实施例1制备的纳米材料在各个器官和肿瘤的含量。
所述两亲性聚合物介导的金纳米组装体使用的浓度为1~20mg/mL,取出的器官有心、肝、脾、肺、肾,以及种植的肿瘤,实验时间点为1~96h。王水硝解时间为1~24h。所述的小鼠模型涉及癌细胞为乳腺癌细胞(MDA-MB-231),而不仅限于乳腺癌模型。
所述的两亲性聚合物介导的金纳米组装体在活体肿瘤成像方面的应用,具体方法为:通过尾静脉往带有皮下瘤的免疫缺陷裸鼠体内注入实施例1制备的纳米材料,然后在不同时间点观察纳米材料在小鼠体内的分布情况。所述两亲性聚合物介导的金纳米组装体使用的浓度为1~20mg/mL,实验时间点为1~144h。所述两亲性聚合物介导的金纳米组装体发射波长范围为400~900nm,激发波长范围为300~650nm,活体肿瘤成像过程观察使用超敏多功能成像仪系统。所述的小鼠模型涉及癌细胞为乳腺癌细胞(MDA-MB-231),而不仅限于乳腺癌模型。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (10)
1.一种两亲性聚合物介导的金纳米组装体的制备方法,其特征在于:包括如下步骤:
1)两亲性聚合物和烷基硫醇在水中形成胶束;加入四氯金酸,混合均匀,获得Au(I)硫醇复合物;所述两亲性聚合物为泊洛沙姆F127;所述烷基硫醇为己硫醇、十二烷基硫醇或十八烷基硫醇中一种以上;
2)调节Au(I)硫醇复合物的pH至碱性,加入还原剂进行还原反应,后续处理,获得两亲性聚合物介导的金纳米组装体;所述还原剂为四羟甲基氯化磷、二甲基胺硼烷、硼氢化钠中一种以上。
2.根据权利要求1所述两亲性聚合物介导的金纳米组装体的制备方法,其特征在于:
所述氯金酸与配体烷基硫醇的摩尔比为1:(1~5);氯金酸与还原剂的摩尔比为(25~220):1。
3.根据权利要求2所述两亲性聚合物介导的金纳米组装体的制备方法,其特征在于:所述氯金酸与配体烷基硫醇的摩尔比为1:(2~4)。
4.根据权利要求1所述两亲性聚合物介导的金纳米组装体的制备方法,其特征在于:所述pH为8~13;
所述烷基硫醇与两亲性聚合物的用量关系为(0.3~1.5)mmol:1g。
5.根据权利要求1所述两亲性聚合物介导的金纳米组装体的制备方法,其特征在于:所述还原反应的时间为3~24h;
步骤1)中所述两亲性聚合物与水的质量比为1:(40~60)。
6.一种由权利要求1~5任一项所述制备方法得到的两亲性聚合物介导的金纳米组装体。
7.根据权利要求6所述两亲性聚合物介导的金纳米组装体,其特征在于:所述的金纳米组装体的长度为10~50nm,宽度为10~25nm;组装体中金纳米单颗粒子粒径为1~3nm。
8.根据权利要求6所述两亲性聚合物介导的金纳米组装体的应用,其特征在于:所述两亲性聚合物介导的金纳米组装体用于医药领域、生物传感器、生物传感检测领域、荧光成像领域以及肿瘤成像的成像剂,以非疾病治疗和非疾病诊断为目的。
9.根据权利要求8所述的应用,其特征在于:所述医药领域为载药领域,用作药物的载体;
所述肿瘤成像的成像剂是指高肿瘤靶向的成像剂。
10.根据权利要求9所述的应用,其特征在于:所述药物为抗肿瘤的药物。
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