CN112870374B - 一种糖响应的功能化纳米复合胶束的制备及在胰岛素递送中的应用 - Google Patents
一种糖响应的功能化纳米复合胶束的制备及在胰岛素递送中的应用 Download PDFInfo
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Abstract
一种糖响应的功能化纳米复合胶束的制备及在胰岛素递送中的应用。三种嵌段共聚物PCL‑b‑PEG、PCL‑b‑P(Asp‑co‑AspPBA)和PCL‑b‑P(Asp‑co‑AspNTA)自组装形成以PCL为核PEG/PAPBA/PANTA为混合壳层的复合胶束。修饰NTA的链段PANTA通过螯合作用结合Zn2+,再利用Zn2+与胰岛素B链上组氨酸的配位作用捕捉胰岛素。具有PBA的链段PAPBA在中性或无糖条件下形成疏水域。被捕捉的胰岛素孵育一段时间,便可绑定到疏水域上,通过配位和疏水的协同作用将胰岛素负载到复合胶束上。在高糖条件下,疏水域由疏水变亲水,复合胶束与胰岛素作用力减弱,胰岛素释放出来进而发挥作用。
Description
技术领域
本发明属于纳米生物医药材料领域,通过复合胶束表面具有的配位域和疏水域负载胰岛素,在高糖环境下发生亲疏水的转变,糖响应性释放胰岛素。
背景技术
糖尿病是一种以糖代谢失调为特征的慢性疾病,由于胰岛素分泌功能障碍或者无法有效利用胰岛素而导致体内血糖升高。近年来,全世界的糖尿病病例迅速增加,并且伴有低龄化的趋势,是危害人类健康的三大杀手之一,因此糖尿病的治疗是一个亟待解决的问题。目前胰岛素作为调节人体葡萄糖代谢的关键激素,是临床上用于治疗1型糖尿病的蛋白类药物,具有起效快的优点。然而患者需要频繁注射外源性胰岛素来控制血糖处于正常水平。这种方式无法模拟出正常人体内内源性胰岛素的释放模式,且会让患者感到不适,易造成注射部位出现炎症和感染等风险。同时,由于难以精准给药,可能会导致患者出现低血糖的症状,甚至导致休克致命。另外,在生产、纯化、储存、运输和递送过程中,胰岛素不能一直保持稳定的结构,它会被外界环境的影响,发生聚集和失活。且临床上也发现,在糖尿病患者反复注射的部位,胰岛素会形成蛋白原纤维,既导致蛋白的生物利用度降低,又严重影响患者体内的血糖控制。目前,已经开发了一系列具有刺激响应性的胰岛素递送体系用于改善胰岛素的治疗效果,如水凝胶,囊泡等。不过,胰岛素存在的易聚集以及稳定性差的问题并未得到很好的解决。故设计出新型的胰岛素递释系统可控释放和保护胰岛素的结构是十分必要的。
发明内容
本发明目的是针对现有技术存在的问题,提供一种通过配位作用和疏水作用将胰岛素捕获并负载在表面的复合胶束的制备方法,通过协同作用既可以提高负载量,又可以保护胰岛素,抑制胰岛素的聚集。同时,在高血糖条件下,疏水域发生从疏水到亲水的转变,负载的胰岛素被释放出来,发挥作用。
本发明的技术方案
一种糖响应的功能化纳米复合胶束,以PCL为核,包括三种嵌段共聚物PCL-b-PEG,PCL-b-P(Asp-co-AspPBA)和PCL-b-P(Asp-co-AspNTA)构成,PEG/PAPBA/PANTA为混合壳层,其PANTA链段捕捉胰岛素,同时糖响应性链段PAPBA在pH 7.4且无糖的条件下形成的疏水域能够通过疏水作用绑定胰岛素,在高血糖下时能够缓慢释放胰岛素。
一种糖响应的功能化纳米复合胶束的制备方法,具体方法如下:
1)PCL-b-PEG的合成
聚乙二醇(PEG-OH)与ε-己内酯(ε-CL)以1:100的摩尔比溶解在适量重蒸甲苯中,先经过液氮冷冻-抽真空-解冻过程,重复三次;在108-112℃条件下反应12h,通过冰乙醚沉淀和真空干燥得到PCL-b-PEG白色粉末;
2)PCL-NHBoc的合成;
小分子引发剂N-(叔丁氧羰基)乙醇胺和ε-CL以1:100的摩尔比溶解在适量重蒸甲苯中,随后加入一滴Sn(Oct)2。冻抽3次之后,放入108-112℃油浴中反应12h。经过冰乙醚沉淀和真空干燥后得到白色固体PCL-NHBoc。
3)PCL-NH2的合成;
将PCL-NHBoc溶于适量三氟乙酸中,室温下搅拌2h。随后通过冰乙醚沉淀和真空干燥得白色固体。再将白色固体溶解在三乙胺和二氯甲烷体积比为1:1的混合溶剂中室温搅拌12h。最后经过冰乙醚沉淀和真空干燥得到大分子引发剂PCL-NH2。
4)BLA-NCA的合成;
称取L-天冬氨酸苄酯在重蒸的四氢呋喃中溶解分散,缓慢升温至60℃。称取三光气加入反应瓶中,其中摩尔体积比L-天冬氨酸苄酯:三光气:四氢呋喃=3mmol:1mmol:12ml,观察到反应液由浑浊变为澄清亮黄色,充氮气除去瓶内过量的光气和氯化氢气体。旋蒸浓缩反应液,正己烷中沉淀抽滤,将抽滤得到的粗产物在乙酸乙酯/正己烷体积比为1:1的混合液中重结晶,抽滤干燥得到BLA-NCA。
5)PCL-b-PAsp(OBzl)的合成;
将PCL-NH2和BLA-NCA以1:50的摩尔比用适量重蒸二氯甲烷溶解分散。冻抽3次后在氮气保护下,于30℃的油浴中反应。经过冰乙醚沉淀和真空干燥后得到白色固体PCL-b-PAsp(OBzl)。
6)PCL-b-PAsp的合成;
PCL-b-PAsp(OBzl)溶解于三氟乙酸中,依次加入苯甲醚和三氟甲磺酸,其中摩尔体积比PCL-b-PAsp(OBzl):三氟乙酸:苯甲醚:三氟甲磺酸=0.01mmol:10ml:1ml:1ml,在冰浴下搅拌1h。反应结束,通过冰乙醚沉淀和真空干燥得到嵌段共聚物PCL-b-PAsp。
7)PCL-b-P(Asp-co-AspPBA)的合成;
PCL-b-PAsp溶于减蒸的DMF中,先加入NHS在室温下搅拌1h。随后依次加入EDC和溶于DMF的氨基苯硼酸PBA,其中摩尔比PCL-b-PAsp:NHS:EDC:PBA=0.01mmol:2.5mmol:1.5mmol:2.5mmol,室温搅拌12h。反应液转移到透析袋中透析3天(MWCO 3500)。冷冻干燥溶液得到黄色固体PCL-b-P(Asp-co-Asp PBA)。
8)PCL-b-P(Asp-co-AspNTA)的合成;
PCL-b-PAsp溶于减蒸的DMF中,先加入NHS在室温下搅拌1h。随后依次加入EDC,和溶于DMF的氨三乙酸要缓慢滴加,其中摩尔比PCL-b-PAsp:NHS:EDC:NTA=0.01mmol:2.5mmol:1.5mmol:2.5mmol,室温搅拌12h。反应液转移到透析袋中3天(MWCO 3500)。冷冻干燥溶液得到白色固体PCL-b-P(Asp-co-AspNTA)。
9)复合胶束的制备;
根据PCL-b-P(Asp-co-AspPBA)在胶束中所占质量比的不同(10%、25%、50%)制备得到不同组成的复合胶束。将成功合成的嵌段共聚物PCL-b-PEG,PCL-b-P(Asp-co-AspPBA)和PCL-b-P(Asp-co-AspNTA)溶解在DMSO中。在剧烈搅拌下快速加入碱性水(pH11.0)到聚合物溶液中。搅拌过夜后混合溶液转移到透析袋(MWCO 3500)中,并在pH 7.4的缓冲液透析1天。然后胶束中再添加醋酸锌溶液,室温搅拌4h,转入透析袋(MWCO 3500)中再透析3天,得到了螯合了锌离子的复合胶束。最后通过称量将溶液定容到0.6mg/mL。
10)胰岛素的负载;
先配制新鲜胰岛素溶液(0.4mg/mL)。然后将胰岛素溶液滴入步骤9)制备的复合胶束溶液(1mL)中,室温搅拌24h。然后在PBS7.4中透析3天去除游离胰岛素,得到负载胰岛素的纳米复合胶束。应用于胰岛素的递送体系中。
本发明提供的糖响应的功能化纳米复合胶束可应用到胰岛素的递送体系中进而治疗I型糖尿病。
本发明的优点和有益效果:
本发明构建了一种糖响应的功能化纳米复合胶束用于胰岛素的投递和保护。该胶束通过配位和疏水的协同作用更稳定地负载胰岛素,保护了胰岛素的结构,抑制其聚集,并实现葡萄糖刺激响应释放。
附图说明
图1为本发明中复合胶束负载和释放胰岛素的过程。
图2为复合胶束CMs的粒径分布。
图3为复合胶束CMs和胰岛素的FRET效应的荧光光谱,用于研究CMs与胰岛素的相互作用。
图4为负载上胰岛素的CMs的荧光光谱,用于计算胰岛素负载量。
图5为负载胰岛素的CMs在不同糖浓度下的释放曲线。
图6为37℃下CMs抑制胰岛素聚集的ThT实验。
图7为复合胶束在体外的细胞毒实验。
具体实施方式
实施例1:
一种糖响应的功能化纳米复合胶束的制备方法,该方法步骤及相关评估如下:
1)PCL-b-PEG的合成
把提前真空干燥过的0.1mmol PEG-OH和减压蒸馏过的10mmolε-CL加到50mL干燥的Schlenk瓶中混合,用7mL重蒸过的无水甲苯溶解,随后加入一滴催化剂辛酸亚锡(Sn(Oct)2)。再通过液氮冷冻—抽真空—解冻,重复三次后,在氮气保护下于108-112℃的油浴中反应12h。反应完毕,加入适量二氯甲烷稀释,然后在十倍体积的冰乙醚中沉淀。放到冰箱静置一夜,再抽滤,洗涤,真空干燥后得到的白色絮状固体即为PCL-b-PEG。
2)PCL-NHBoc的合成;
干燥的小分子引发剂0.1mmol N-(叔丁氧羰基)乙醇胺和减压蒸馏过的10mmolε-CL加到50mL干燥的Schlenk瓶中混合,用7mL重蒸过的无水甲苯溶解,随后加入一滴催化剂Sn(Oct)2。再通过液氮冷冻—抽真空—解冻,重复三次后,在氮气保护下于108-112℃的油浴中反应12h。反应完毕,加入适量二氯甲烷稀释,然后在十倍体积的冰乙醚中沉淀。放到冰箱静置过夜,再抽滤,洗涤,真空干燥后得到的白色絮状固体PCL-NHBoc。
3)PCL-NH2的合成;
将0.1mmol PCL-NHBoc放到干燥的50mL烧瓶中,加入5ml三氟乙酸溶解,室温下搅拌2h。随后在十倍体积的冰乙醚中沉淀,待沉淀完全,再抽滤,洗涤真空干燥得白色固体。再将0.1mmol白色固体放到干燥的50mL烧瓶,用10mL三乙胺和二氯甲烷体积比为1:1的混合溶剂溶解,室温搅拌12h。最后经过冰乙醚沉淀和真空干燥得到大分子引发剂PCL-NH2。
4)BLA-NCA的合成;
称取30mmol L-天冬氨酸苄酯置于具有尾气吸收装置的干燥的500ml反应瓶中,加入120mL重蒸的四氢呋喃溶解分散,缓慢升温至60℃。称取10mmol三光气加入反应瓶中,观察到反应液由浑浊变为澄清亮黄色,充氮气除去瓶内过量的光气和氯化氢气体。旋蒸浓缩反应液,在10倍体积的正己烷中沉淀,沉淀完全再抽滤。将抽滤得到的粗产物先用100mL乙酸乙酯在500mL烧杯中溶解,随后加入100mL正己烷进行重结晶,静置过夜,抽滤干燥得到BLA-NCA。
5)PCL-b-PAsp(OBzl)的合成;
将0.1mmol PCL-NH2和5mmol BLA-NCA放到50mL干燥的Schlenk瓶中混合,用15ml重蒸二氯甲烷溶解分散,冻抽3次后在氮气保护下于30℃的油浴中反应。当反应液由浑浊变透明说明反应完全,在10倍体积的冰乙醚中沉淀,放到冰箱静置过夜,再抽滤,洗涤,真空干燥后得到白色固体PCL-b-PAsp(OBzl)。
6)PCL-b-PAsp的合成;
将0.01mmol PCL-b-PAsp(OBzl)放到50mL干燥的圆底烧瓶中,用10mL三氟乙酸溶解,随后依次加入1ml苯甲醚和1ml三氟甲磺酸,在0℃冰浴下搅拌1h。反应结束,在10倍体积的冰乙醚中沉淀,放到冰箱静置过夜,再抽滤,洗涤,真空干燥后得到白色固体PCL-b-PAsp。
7)PCL-b-P(Asp-co-AspPBA)的合成;
将0.01mmol PCL-b-PAsp放到50mL干燥的圆底烧瓶中,用10mL减蒸过的DMF溶解。先加入2.5mmol NHS在室温下搅拌1h,随后先加入1.5mmol EDC,充分溶解后再加入溶于DMF的2.5mmol氨基苯硼酸PBA,室温搅拌12h。反应结束后,反应液转移到透析袋(MWCO 3500)中透析3天。然后,将反应液放到-80℃的冰箱冷冻4h,再放到冷冻干燥机干燥,最后得到黄色固体PCL-b-P(Asp-co-Asp PBA)。
8)PCL-b-P(Asp-co-AspNTA)的合成;
将0.01mmol PCL-b-PAsp放到50mL干燥的圆底烧瓶中,用10mL减蒸过的DMF溶解。先加入2.5mmol NHS在室温下搅拌1h,随后先加入1.5mmol EDC,充分溶解后,再缓慢滴加溶于DMF的2.5mmol氨三乙酸(加一滴三乙胺助溶),室温搅拌12h。反应结束后,反应液转移到透析袋(MWCO 3500)中透析3天。然后,将反应液放到-80℃的冰箱冷冻4h,再放到冷冻干燥机干燥,最后得到白色固体PCL-b-P(Asp-co-AspNTA)。
9)不同比例的复合胶束的制备;
a)质量含量为10%的复合胶束的制备
称取2.7mg PCL-b-PEG,0.6mg PCL-b-P(Asp-co-AspPBA)和2.7mg PCL-b-P(Asp-co-AspNTA)溶解在1mL DMSO中。在剧烈搅拌下快速加入8mL碱性水(pH 11.0)到聚合物溶液中。搅拌过夜后混合溶液转移到透析袋(MWCO 3500)中,并在pH 7.4的缓冲液透析1天。然后向胶束中再添加10μL醋酸锌溶液(1mg/mL),室温搅拌4h,转移到透析袋(MWCO 3500)中再透析3天,得到PCL-b-P(Asp-co-AspPBA)质量含量为10%的复合胶束。最后通过称量将溶液定容到0.6mg/mL。
b)质量含量为25%的复合胶束的制备
称取2.25mg PCL-b-PEG,1.5mg PCL-b-P(Asp-co-AspPBA)和2.25mg PCL-b-P(Asp-co-AspNTA)溶解在1mL DMSO中。在剧烈搅拌下快速加入8mL碱性水(pH 11.0)到聚合物溶液中。搅拌过夜后混合溶液转移到透析袋(MWCO 3500)中,并在pH 7.4的缓冲液透析1天。然后向胶束中再添加25μL醋酸锌溶液(1mg/mL),室温搅拌4h,转移到透析袋(MWCO3500)中再透析3天,得到PCL-b-P(Asp-co-AspPBA)含量为25%的复合胶束。最后通过称量将溶液定容到0.6mg/mL。
c)质量含量为50%的复合胶束的制备
称取1.5mg PCL-b-PEG,3mg PCL-b-P(Asp-co-AspPBA)和1.5mg PCL-b-P(Asp-co-AspNTA)溶解在1mL DMSO中。在剧烈搅拌下快速加入8mL碱性水(pH 11.0)到聚合物溶液中。搅拌过夜后混合溶液转移到透析袋(MWCO 3500)中,并在pH 7.4的缓冲液透析1天。然后向胶束中再添加50μL醋酸锌溶液(1mg/mL),室温搅拌4h,转移到透析袋(MWCO 3500)中再透析3天,得到PCL-b-P(Asp-co-AspPBA)含量为50%的复合胶束。最后通过称量将溶液定容到0.6mg/mL,并将该胶束应用于其他实例中。如图2所示,通过动态光散射(DLS)表征复合胶束的粒径,平均粒径为115nm。
10)复合胶束负载和释放胰岛素的过程
如图1所示,聚合物自组装形成复合胶束,PEG和PANTA链段伸展,有PBA基团的链段PAPBA在pH7.4条件下形成疏水域。该复合胶束先通过螯合作用结合Zn2+,再利用Zn2+与胰岛素B链上组氨酸残基的配位作用捕捉胰岛素。被捕捉的胰岛素经过一段时间的孵育,便可绑定到疏水域,这样复合胶束便成功负载上了胰岛素。在高糖条件下,疏水域由疏水变亲水,复合胶束与胰岛素作用力减弱,胰岛素释放出来,发挥作用。
实施例2:糖响应性功能化聚合物胶束的应用。
一种糖响应的功能化纳米复合胶束的应用,该方法步骤及相关评估如下:
1)复合胶束与胰岛素的相互作用的研究;
通过FRET效应研究合胶束与胰岛素的相互作用。其原理是当供体和受体两者距离足够近(一般小于10nm),激发荧光供体后,供体激发态能量会非放射性转移到受体,诱导激发荧光受体,发生能量的转移。在本研究中,将RhB标记到复合胶束的疏水域上,并取1mL该胶束与0.2mL FITC标记的胰岛素在37℃下混合2h,记录荧光光谱(FITC是供体,RhB是受体,激发波长450nm)。如图3所示,RhB-CMs和FITC-Insulin孵育后,出现两个特征峰(525nm,585nm),说明胰岛素成功负载到胶束上。
2)胰岛素的负载;
先配制新鲜FITC标记的胰岛素溶液(0.4mg/mL),避光保存好。然后向有复合胶束溶液(1mL)的小瓶中,添加0.2mL胰岛素溶液,室温搅拌24h。然后在PBS7.4中透析3天去除游离胰岛素,最后得到负载胰岛素的纳米复合胶束。配置了7个不同浓度的FITC标记的胰岛素溶液(0.01、0.02、0.05、0.1、0.2、0.5、1.0mg/mL),用荧光光度计得到其荧光光谱,记录在525nm处的荧光强度,得到FITC-Insulin的标曲。如图4所示,在520nm处的荧光发射峰既说明FITC-INS已经负载到复合胶束上,又可以通过荧光强度值计算负载量。
3)胰岛素的释放
将1mL载有FITC-胰岛素复合胶束溶液注射到分子量截止值为12kDa的透析袋中,并浸泡在13mL葡萄糖浓度分别为0、1、和4g/L的PBS 7.4的溶液中。每隔一段时间,取样1ml透析液以测量FITC-insulin的浓度,并加入1m新鲜缓冲液。如图5所示,在4g/L的浓度下释放得快;0g/L和1g/L浓度下释放得慢,说明释放行为具有葡萄糖响应性。
4)复合胶束抑制胰岛素聚集;
通过将胰岛素溶液(0.4g/L)分别与PBS 7.4和复合胶束混合得到ThT样品,最终样品中胰岛素含量0.2mg/ml,复合胶束含量0.6mg/ml。在37℃下持续搅拌(800rpm)孵育12h,每隔一段时间取出50μL样品,并与450μLThT缓冲溶液混合作为测试样品。记录在440nm激发波长下485nm处的荧光发射强度。如图6所示,胰岛素溶液(Ins/PBS)的荧光强度明显增加,而Ins/CMs的荧光强度很低,说明复合胶束有抑制胰岛素聚集的作用。
5)复合胶束的安全性评估;
复合胶束的细胞毒性选用NIH 3T3小鼠成纤维细胞,且通过CCK8法进行评估。设置5个不同浓度的复合胶束溶液(1、10、100、500、1000μg/mL)。如图7所示,各个浓度下,细胞粗活率都在90%以上,证明复合胶束无毒,有较好的生物相容性。
Claims (4)
1.一种糖响应的功能化纳米复合胶束的制备方法,其特征在于步骤如下:
1)PCL-b-PEG的合成;
聚乙二醇PEG-OH与ε-己内酯ε-CL以1:100的摩尔比溶解在重蒸甲苯中,先经过液氮冷冻-抽真空-解冻过程,重复;在108-112℃条件下反应12h,通过冰乙醚沉淀和真空干燥得到PCL-b-PEG白色粉末;
2)PCL-NHBoc的合成;
N-(叔丁氧羰基)乙醇胺和ε-CL以1:100的摩尔比溶解在重蒸甲苯中,随后加入一滴Sn(Oct)2,冻抽3次之后,放入108-112℃油浴中反应12h,冰乙醚沉淀和真空干燥后得到白色固体PCL-NHBoc;
3)PCL-NH2的合成;
将PCL-NHBoc溶于三氟乙酸中,室温下搅拌2h,冰乙醚沉淀和真空干燥得白色固体,将白色固体溶解在体积比为1:1的三乙胺和二氯甲烷的混合溶剂中室温搅拌12h,冰乙醚沉淀和真空干燥得到PCL-NH2;
4)BLA-NCA的合成;
称取L-天冬氨酸苄酯在重蒸的四氢呋喃中溶解分散,缓慢升温至60℃,称取三光气加入反应瓶中,其中摩尔体积比L-天冬氨酸苄酯:三光气:四氢呋喃=3mmol:1mmol:12ml,观察到反应液由浑浊变为澄清亮黄色,充氮气除去瓶内过量的光气和氯化氢气体,旋蒸浓缩反应液,正己烷中沉淀抽滤,将抽滤得到的粗产物在乙酸乙酯/正己烷体积比为1:1的混合液中重结晶,抽滤干燥得到BLA-NCA;
5)PCL-b-PAsp(OBzl)的合成;
将PCL-NH2和BLA-NCA以1:50的摩尔比用重蒸二氯甲烷溶解,冻抽3次,在氮气保护下,于30℃的油浴中反应,冰乙醚沉淀和真空干燥后得到白色固体PCL-b-PAsp(OBzl);
6)PCL-b-PAsp的合成;
PCL-b-PAsp(OBzl)溶解于三氟乙酸中,依次加入苯甲醚和三氟甲磺酸,其中摩尔体积比PCL-b-PAsp(OBzl):三氟乙酸:苯甲醚:三氟甲磺酸=0.01mmol:10ml:1ml:1ml,在冰浴下搅拌1h,冰乙醚沉淀和真空干燥得到嵌段共聚物PCL-b-PAsp;
7)PCL-b-P(Asp-co-AspPBA)的合成;
PCL-b-PAsp溶于减蒸DMF中,先加入NHS在室温下搅拌1h,依次加入EDC,溶于DMF的氨基苯硼酸PBA,其中摩尔比PCL-b-PAsp:NHS:EDC:PBA=0.01mmol:2.5mmol:1.5mmol:2.5mmol,室温搅拌12h,放入透析袋透析3天,冷冻干燥溶液得到黄色固体PCL-b-P(Asp-co-AspPBA);
8)PCL-b-P(Asp-co-AspNTA)的合成;
PCL-b-PAsp溶于减蒸DMF中,先加入NHS在室温下搅拌1h,依次加入EDC,溶于DMF的氨三乙酸要缓慢滴加,其中摩尔比PCL-b-PAsp:NHS:EDC:NTA=0.01mmol:2.5mmol:1.5mmol:2.5mmol,室温搅拌12h,放入透析袋透析3天,冷冻干燥溶液得到白色固体PCL-b-P(Asp-co-AspNTA);
9)复合胶束的制备;
根据PCL-b-P(Asp-co-AspPBA)在胶束中所占质量比分别按照10%、25%和50%制备得到不同组成的复合胶束,将步骤1)得到的PCL-b-PEG,步骤7)得到的PCL-b-P(Asp-co-AspPBA)和步骤8)得到的PCL-b-P(Asp-co-AspNTA)溶解在DMSO中,在剧烈搅拌下快速加入pH为11.0的碱性水到聚合物溶液中,搅拌过夜,转入透析袋中在pH 7.4的缓冲液中透析1天,然后胶束中再添加醋酸锌溶液,室温搅拌4h,转入透析袋中再透析3天,得到螯合了锌离子的复合胶束,最后通过称量将溶液定容。
2.权利要求1所述方法制备的糖响应的功能化纳米复合胶束。
3.权利要求1所述方法制备的糖响应的功能化纳米复合胶束在制备胰岛素的递送体系中的应用。
4.根据权利要求3的应用,其特征在于,包括胰岛素的负载;
先配制新鲜的0.4mg/mL胰岛素溶液,然后将胰岛素溶液滴入权利要求1所述的复合胶束溶液中,搅拌24h,然后在PBS7.4中透析3天去除游离胰岛素,得到胰岛素的递送体系即负载胰岛素的纳米复合胶束。
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