CN114948784A - 一种仿生纳米颗粒及其制备方法和用途 - Google Patents
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Abstract
本发明属于生物医药技术领域，公开了一种仿生纳米颗粒及其制备方法和用途。该制备方法包括以下操作步骤：将含有二硫键和/或巯基的蛋白、多肽或多糖在35～60℃温度条件下，采用二硫键还原剂进行处理，经过诱导组装，分离，得到仿生复合纳米颗粒。实验表明，跟单体(蛋白或多肽或多糖)比较，本发明仿生纳米颗粒具有制备简单、稳定性好、活性保存好和安全性更高等特点，为仿生纳米颗粒制备提供了新方案，可很好的用于制备抗皮肤衰老、创面修复或药物递送产品。
Description
技术领域
本发明属于生物医药技术领域，特别涉及一种仿生纳米颗粒及其制备方法和用途。
背景技术
近年来，随着纳米技术的不断发展和产业成熟，如脂质体抗癌药物、紫杉醇白蛋白结合纳米药物、四氧化三铁纳米药物等的上市推广，大量研究发现纳米尺度对物质颗粒赋予了一些新的理化和生物功能作用。其中，四氧化三铁纳米颗粒等还具备了内在的生物酶活性，且开发成本低、易于化学改造、稳定性好等优点。目前，随着纳米颗粒等在肿瘤动力学治疗上引起高度关注之外，其在医药、化工、食品、农业和环境等领域亦具有广大的应用前景。因此，通过绿色制备手段，构建新的仿生纳米颗粒，在科学基础和应用方面均具有非常重大的意义。
近年来，关于蛋白纳米颗粒制备的工艺有不少，如2012年，高会乐等(CN102626393B)将白蛋白和难溶性替尼类药物结合，用磷脂分散和稳定，构建了可溶性注射用白蛋白纳米颗粒；2012年，高缘等(CN201415999A)将戊二醛固化的白蛋白纳米粒吸附灯盏花素制备纳米药物；2013年，张晓宏等(CN103169968B)通过白蛋白物理吸附疏水性二氢卟吩光敏剂获得纳米光敏剂；闫学海等(CN103520734B)通过带正电荷的大分子和白蛋白分子静电吸附及还原剂作用下构建了白蛋白纳米粒子；邓意辉等(CN1919339B)用50～99.9％(W/W)的白蛋白及其他辅料，在高压均质仪作用下，制备了葫芦素-白蛋白固体制剂；张文芳等(CN103908430A)采用表面稳定剂和白蛋白混合，高压均化法制备得到可稳定存放12小时以上的紫杉醇纳米粒；陈道桢等(CN103768024B)采用反溶剂和戊二醛交联方法制备了人参皂苷Rh2-白蛋白纳米粒；王文坦等(CN103212083B)通过用醇类等反溶剂沉淀白蛋白，得到分子间二硫键重组的白蛋白纳米颗粒，其可在稀释条件下稳定及还原环境中响应释放；2014年，蔡林涛等(CN104189916B、CN104162164B)通过将白蛋白二硫键还原后，在反溶剂(乙醇)作用下得到分子之间含巯基和/或二硫键的多聚体白蛋白纳米球；2015年，李芳等(CN104490847A)通过加入香草醛类似物在加热条件将白蛋白分子内游离巯基形成分子内二硫键，蛋白上的氨基和香草醛的醛基形成席夫碱等化学键，得到水溶液中稳定的纳米颗粒；姜虎林等(CN105288647A)通过将白蛋白修饰点击反应功能化，在金属离子和药物的配位作用下构建了白蛋白纳米颗粒；2016年，张强等(CN105816885A)采用亲和素和白蛋白之间的静电相互作用，构建了亲和素白蛋白纳米粒子，可靶向经生物素偶联抗体富集的肿瘤；钟延强等(CN105796502A)通过类似方法制备了蓝萼甲素白蛋白纳米粒；姜虎林等(CN105879045)通过药物和载体白蛋白间的静电吸附和配位作用，及反溶剂作用下将载体自身氨基酸残基的交联实现了抗肿瘤药物阿霉素和造影剂MnO2胶体纳米粒的共载白蛋白纳米粒；黄永焯等(CN107157950A)将白蛋白溶解于脲，通过NaBH4变性蛋白后，加入反溶剂(乙醇)沉淀得到白蛋白纳米颗粒；黄海等(CN106540270A)通过二硫键的还原-氧化反应获得了紫杉醇和全反式维甲酸共载纳米颗粒；2017年，陈立江等(CN107126564A)通过类似Abraxane工艺的高速高压均质法制备得白蛋白结合型索拉菲尼(Abraxane工艺被证明并非二硫键结合稳定(Biomacromolecules,2012；13:23-28.))；2019年，杨鹏等(CN110563979A)在4-羟乙基哌嗪乙磺酸溶液中基于巯基与二硫键交换反应制备得到蛋白质纳米薄膜；2020年，蔡宇等通过乙醇溶剂、超声和滴加戊二醛溶液进行交联固化白蛋白，公开了一种包载黄芩苷的叶酸偶联白蛋白纳米粒的制备方法(CN111249254B)。上述这些制备方法以载药技术问题为核心，但绝大部分需要分步引入大量的反溶剂(乙醇、甲醇等)和交联剂进行固定化反应(戊二醛等)，对制备技术带来了分离难度和高成本。同时，尚未见到有同时制备大分子自身重构的工艺及其功能评价。
鉴于此，申请人前期通过对含巯基或二硫键的多糖、多肽或蛋白质的重构工艺进行研究，发现其构建的仿生纳米颗粒具有清除自由基或超氧根及促进皮肤修复等生物活性性质，在皮肤衰老、创面修复和药物递送方面具有良好的应用前景。
发明内容
为了解决现有技术存在的缺点和不足，本发明的首要目的在于提供一种仿生纳米颗粒的制备方法，该方法操作简单，将含有二硫键和/或巯基的蛋白、多肽或多糖在35～60℃温度条件下，采用二硫键还原剂进行处理，经过诱导组装，分离，即可得到尺寸均一，分散性好，可室温保存及在体液中稳定的新型仿生纳米颗粒。
本发明的另一目的在于提供一种上述制备方法制备得到的仿生纳米颗粒。
本发明的再一目的在于提供一种上述仿生纳米颗粒的用途。
本发明的目的通过下述技术方案来实现：
一种仿生纳米颗粒的制备方法，包括以下操作步骤：将含有二硫键和/或巯基的蛋白、多肽或多糖在35～60℃温度条件下与二硫键还原剂进行共同处理，经过诱导组装，分离，得到仿生纳米颗粒。
所述含有二硫键和/或巯基的蛋白、多肽或多糖包括角蛋白、牛奶蛋白、酪蛋白、鱼皮蛋白、玉米蛋白、乳清蛋白、乳蛋白、蚕丝胶蛋白、乳球蛋白、豆浆蛋白、小麦蛋白、大豆蛋白、燕麦蛋白、乳铁蛋白、胶原蛋白、血清蛋白、麦醇溶蛋白、弹性蛋白、人纤维蛋白(原)、胎盘蛋白、透明质酸、伸展蛋白、肌动蛋白、酵母蛋白、马铃薯蛋白、大米蛋白、米糠蛋白、木瓜蛋白酶、网硬蛋白、小麦谷蛋白、羽扇豆蛋白、芝麻蛋白、榛子蛋白、可溶性蛋白多糖或者其水解产物及修饰产物中的一种以上。
所述二硫键还原剂为半胱氨酸、二硫苏糖醇、还原型谷胱甘肽、β-巯基乙醇或三(2-羧乙基)膦。
上述的一种仿生纳米颗粒的制备方法，具体包括如下步骤：将含有二硫键和/或巯基的蛋白、多肽或多糖配制成质量分数0.1％～20％的溶液，加热升温至35～60℃，再加入浓度为0.1～200mM的二硫键还原剂水溶液，pH值调节至5.0～9.0，搅拌2～24小时，制备得到纳米颗粒溶液；再经过透析或离心，冻干，得到仿生纳米颗粒。
所述透析是将纳米颗粒溶液放入透析袋，并于0～30℃的等渗缓冲溶液或纯水中透析除去多余的二硫键还原剂及其副产物；透析分子截留不低于1000；所述离心的转速为8000～15000rpm，10分钟/次，离心的次数为1～3次。
一种由上述的制备方法制备得到的仿生纳米颗粒，该仿生纳米颗粒由分子间巯基和/或二硫键重新装配成球状纳米结构。
所述仿生纳米颗粒在常温情况下，在水溶液中和各种体液中均可稳定存在。
上述的仿生纳米颗粒在制备抗皮肤衰老产品、创面修复产品或药物递送产品中的用途。
上述仿生纳米颗粒在使用的时候可以制成、口服制剂和非口服制剂。
本发明的原理：本发明将含有二硫键和/或巯基的蛋白、多肽或多糖，于35～60℃环境下，在二硫键还原剂作用下，打开其内部结构，舒展开巯基等活性基团，重新让蛋白质区域互相吸引，诱发分子间的巯基/二硫键等的重构，获得稳定性优异的新型仿生纳米颗粒。
本发明相对于现有技术，具有如下的优点及有益效果：
本发明将含有二硫键和/或巯基的蛋白或多肽或多糖，在二硫键还原剂作用下诱发分子间的巯基/二硫键等重构，即改性后分子间相互作用和重构大分子组成，成球状纳米结构，获得稳定性优异的新型仿生纳米颗粒，在机体组织穿透、融合和修复及稳定性保存方面比原型具有更优的能力；功能研究表明，该仿生纳米颗粒在皮肤衰老、创面修复和药物递送方面具有优异功能。
附图说明
图1为AC NPs的DLS水合粒径图。
图2为AC NPs的TEM电镜图。
图3为AC NPs的仿生酶活性评价图。
图4为AC NPs对正常皮肤细胞毒性影响图。
图5为AC NPs对辐射胶原细胞HaCaT的生存率影响图。
图6为AC NPs对辐射胶原细胞HaCaT的增殖率影响图。
图7为AC NPs对辐射小鼠皮肤胶原组织影响图。
图8为AC NPs对辐射小鼠皮肤的HE影响图。
图9为AC NPs对辐射小鼠皮肤胶原蛋白代谢因子Col1a1、Col3a3和MMP1的mRNA影响图。
图10为AC NPs对辐射小鼠皮肤炎症因子TNF-α和IL-1β的mRNA影响图。
图11为AC NPs对辐射小鼠皮肤氧化应激信号Nrf2和Gpx1的mRNA影响图。
图12为AC NPs对辐射小鼠皮肤衰老标志物P16和P21的mRNA影响图。
图13为AC NPs对辐射小鼠皮肤抗氧化蛋白SOD和MDA表达影响图。
具体实施方式
下面结合实施例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。
实施例1仿生纳米颗粒(AC NPs)的制备：
将牛奶蛋白溶于pH 7.0的生理盐水溶液中配制成质量分数5％的溶液，于35℃水浴中，再加半胱氨酸的水溶液(100mM)，pH值调节至7.0，搅拌6小时，制备得到纳米颗粒溶液；再经过透析(透析袋截留的分子质量，MW＝8000～14000，透析48小时，6小时/次)或离心(10，000rpm，15分钟*3次)，冻干，得到纳米尺寸和分布均匀的仿生纳米颗粒(AC NPs)，平均粒径约20nm(如图1和图2所示)。
实施例2～8仿生复合纳米颗粒(AC NPs)的制备：
参照实施例1制备流程，在实施例2～8中，我们进行了制备条件的调整，其中还原剂在实施例4，5和7用还原型谷胱甘肽，实施例2，3和6用二硫苏糖醇，实施例8用半胱氨酸；其余条件与实施例1一致。
表1：实施例2～8中各制备条件
实施例9游离巯基测定表征仿生纳米颗粒(AC NPs)的重建：
为了证明AC NPs在含巯基分子重建前后的二硫键结合形态，申请人通过测定前后的自由巯基的改变来确证(原型分子(P)，反应的原型分子(RP)和重建后的AC NPs)，采用Ellment的分光光度法检测(AnalBioanal Chem.2002；373(4-5):266-76.)。(1)Ellman试剂制备(10mM DTNB(5,5′-dithio-bis(2-nitrobenzoic acid；5，5'-二硫代-2-硝基苯甲酸)，±10mM Cystamine(胱氨酸，用于建立标准曲线))，即取77.1mg的DTNB直接或添加56.3mg胱氨酸二盐酸盐(Cystamine dihydrochloride)溶于pH7.0的缓冲液中，当所有DTNB溶解后，重调pH至7.0，将其冻存于-20度，解冻后试剂应该当天用完。(2)将0.42mLHCl(37％,4.5mmol)和10mL水溶液加入181.1mg的半胱氨酸(1.5mmol)粉末中，搅拌均匀，然后用水稀释至500ml至3mM终浓度(应1-2小时内用完)。取3mL的PBS和0、10、20、30或40μL的3mM半胱氨酸标准溶液混合，加入50μL Ellman缓冲液(10mM，7.0)，室温反应5min后，在412nm波长进行酶标板测定，建立标准曲线。(3)用PBS将测定巯基样品的理论浓度低于40μM，吸出1mL，加入200μL的强缓冲液(pH 8.2，100mM硼酸和0.2mM EDTA，NaOH调节pH至8.2)调节其pH值≥8.0；然后，加入20μL的Ellman/cystamine试剂，立即涡旋，室温反应5min，在412nm波长下面检测(1cm检测池)A412s；同时，1mL的PBS混合200μL的强缓冲液(pH 8.2)及20μL的Ellman/cystamine试剂进行平行溶剂对照测定A412rp；1mL的测定样品溶液混合200μL的强缓冲液(pH8.2)及20μL的Ellman/cystamine试剂进行平行样品对照的测定A412p。具体计算按：Δε412＝14,150M–1cm–1；SH(mol)＝0.00122L×(A412s-A412rp-A412p)/(Δε412×1cm)，然后根据样品的摩尔量，计算其相对改变值，如表2所示。研究结果表明，实施例1所制备的AC NPs是从原型分子(牛奶蛋白P)经历二硫键的切开(RP)和分子间重组后得到的蛋白质纳米颗粒。
表2：实施例1制备AC NPs的自由巯基改变
实施例10仿生纳米颗粒(AC NPs)的圆二色谱分析：
为进一步证明AC NPs在重建前后的组装形态，对实施例1制备的样品，申请人通过圆二色谱仪测定其分子多维改变来确证(原型分子(P)，和重建后的AC NPs)，研究结果发现，在198-250nm的二级结构特征峰区间，分子间重建后的蛋白质颗粒AC NPs的圆二色谱与原型分子P基本一致，我们参照文献(ACSNano.2014；8(12):12310-22.的附录Figure S2)比对，提示在分子间二硫键重构后，并没显著影响其二级结构。
实施例11仿生纳米颗粒(AC NPs)的表征和鉴定：
采用动态光散射仪DLS，对前述实施例1获得的AC NPs分别进行测定其水合粒径，从结果来看，水合粒径～100nm，PDI<0.40，提示其在水溶液中的分散性良好(如图1)。同时，对其进行了透射电子显微镜TEM观察，其脱水后的干燥粒径约20nm左右(如图2)，显示为球状纳米结构。
实施例12仿生纳米颗粒(AC NPs)的仿生酶活性评价：
为了评价AC NPs(实施例1所得)的仿生酶活性，将200μL的AC NPs(400μg/mL)或牛奶蛋白(Mix)或去离子水对照加入到10mL的离心管中(3.7mL的醋酸钠去离子水溶液(0.2mol/L，pH 4.0)，混合均匀，再加入100μL的3,3',5,5'-四甲基联苯胺(TMB，0.1mM)，涡旋震荡充分；将反应管于室温下反应60分钟，每隔10分钟在652nm处检测反应溶液的吸光值。测定结果表明，AC NPs具有仿生酶特性，而水解胶原蛋白肽或壳多糖几乎没有仿生酶效应(如图3)。
实施例13仿生纳米颗粒(AC NPs)的保存稳定性考察：
采用DLS测定仪对实施例3所得AC NPs纳米颗粒的水合粒径尺寸和分散性进行测定，结果表明其可以较长时间保持稳定(表3和表4)。
表3不同保存时间下AC NPs颗粒分散性观察
表4不同保存时间下AC NPs粒径变化(平均值±SD)
实施例14仿生纳米颗粒(AC NPs)在模拟体液中的稳定性情况：
按着国家药典标准，配制模拟胃液、肠液和临床上常用的替代血浆组分样品。取0.5mL实施例4制备得到的AC NPs颗粒，用模拟液稀释到5mL体积，移液器将其加入微透析袋中，在1000mL体积的模拟体液中于37℃恒温搅拌不同时间，移液器取样测定AC NPs水合粒径，结果表明AC NPs可以在模拟体液中比较稳定保持(表5)。
表5AC NPs在模拟体液中的稳定性测定(平均值±SD)
实施例15 AC NPs修复皮肤衰老的疗效评价：
细胞水平：先将实施例2制备的AC NPs及其原型乳蛋白(Mix组分)对正常皮肤细胞L929(中国科学院上海细胞库)进行安全性剂量考察；然后，选择10～1000μg/mL范围内的ACNPs及Mix组分与HaCaT细胞(中国科学院上海细胞库)于96孔板分别共培养12个小时后，采用紫外灯照射细胞层面(紫外灯选择311nm波长，灯与96孔板液面16cm距离，去96孔板盖子，辐照30s，按80mJ/cm2的能量辐射)，然后继续孵育24和48小时，采用CCK8等分别评价HaCaT细胞的生存率、细胞增殖率和胶原细胞的生存活力。
动物水平：将已适应生存的8周龄SPF级ICR小鼠(购买于斯莱克动物有限公司)，每组8只，先用戊巴比妥麻醉，然后刮毛器脱毛，将0.3mL无菌的的AC NPs(500μg/mL)、Mix组分(同等物质浓度乳蛋白的无菌生理盐水溶液)和生理盐水在圈定皮下进行注射(～2cm2)，在紫外灯(340nm波长)，距离皮肤20cm远，按8W的功率，每次辐射50s，每周3次，接连10个周，诱导出小鼠皮肤光老化模型和评价干预的疗效。将小鼠麻醉，牺牲，用皮肤取样器将Mark笔圈定的皮肤组织进行取样、高分辨拍照，做基因、免疫组化分析与Masson染色，评价胶原纤维蛋白和相关蛋白表达水平(mRNA:Col1a1、Col3a3和MMP1(胶原蛋白代谢因子)；TNF-α和IL-1β(炎症因子)；Nrf2和Gpx1(氧化应激途径)；P16和P21(衰老标志蛋白)；Actin(蛋白对照)；SOD和MDA(抗氧化蛋白))。研究结果表明：细胞水平来说，AC NPs对正常皮肤细胞无任何毒性；与Mix组比较，对紫外辐射HaCaT胶原细胞的生存率、生长增殖率和生存活力均显著增强；在动物水平上，AC NPs组与对照组ICR小鼠皮肤水平一致(光滑度、光泽性、嫩度)，Mix组也具有较好的皮肤防护作用；在mRNA基因调控与蛋白分子水平，AC NPs组可以抵抗紫外灯辐射产生的光老化皮肤损伤，其具体机理与AC NPs可以更好地修复胶原蛋白、减少炎症、降低氧化应激和降低衰老蛋白表达有直接关系；且AC NPs组比Mix组的修复效果更好，这与ACNPs具有更好仿生酶活性有关。上述结果证明AC NPs具有优异的抗皮肤老化和修复效应(如图4～13)。
实施例16 AC NPs卡波姆凝胶对糖尿病大鼠背部创面修复作用
糖尿病大鼠创面模型：每组10只SD大鼠(雌/雄性各半，200g左右)，尾静脉注射链脲佐霉素(65mg/kg剂量)，先造糖尿病大鼠进行造模。于5，10，15天分别从大鼠的眼眶用肝素抗凝毛细管进行取血，测定血糖，以>16.7mmol/L血糖浓度为模型标准。将糖尿病模型成功的大鼠，戊巴比妥腹腔注射麻醉，电动刮毛刀脱毛；待刮毛处颜色和表层恢复正常后，用皮肤取样器从大鼠背部剪下2x1 cm的皮肤，即为糖尿病大鼠创面模型。分别给予生理盐水、AC NPs卡波姆凝胶(将实施例2制备好的AC NPs(300μg/mL)和卡波姆P934(1％质量比)搅拌，滴加三乙醇胺，充分搅拌即可制备获得)和乳蛋白Mix组分卡波姆凝胶(将同等质量的乳蛋白，按AC NPs卡波姆凝胶工艺获得)，按每天1次涂抹，连续给药2周；然后，通过表观拍照，计算创面愈合率(将愈合创面/原始创面*100％)进行计算，结果证明AC NPs具有更好促进创面愈合的作用(表6)。
表6 AC NPs对糖尿病大鼠创面修复的作用(n＝10)
注：与对照组比较：**p<0.01。
实施例17 AC NPs对放射性大鼠模型皮损的修复评价：
将9周龄的雄性SD大鼠(200g左右，每组10只)，腹腔注射苯戊巴比妥麻醉后，放射专用标记Mark笔圈定部位(4cm x 5cm)，采用瓦里安23EX直线加速器照射皮肤(6MEV电子线)，按50Gy的总计剂量进行放疗；每天用无菌棉签涂抹AC NPs卡波姆凝胶(实施例8，ACNPs(300μg/mL)和1％卡波姆P934进行室温搅拌，滴加三乙醇胺后，充分搅拌制备获得。)或水解燕麦蛋白Mix组分卡波姆凝胶组(将同等质量的水解燕麦蛋白，按AC NPs卡波姆凝胶工艺获得)于辐射损伤皮肤，连续5周，拍照，收集采样图进行计算。结果证明：AC NPs组SD大鼠辐射皮肤完整愈合率为100％，肤质红润，光泽发亮，与正常组大鼠的皮肤一致，且毛囊毛发正常生长；而水解燕麦蛋白Mix组辐射皮肤部位的完整修复率为10％，仍然伴有紫红色炎症斑点，其剩余9只大鼠辐射部位皮肤的康复率均不超过60％，其余疤痕部位均有结块，下肉红色；这些结果充分证明，AC NPs具有更好地修复放射性皮肤损伤。
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 (7)
1.一种仿生纳米颗粒的制备方法，其特征在于包括以下操作步骤：将含有二硫键和/或巯基的蛋白、多肽或多糖在35～60℃温度条件下与二硫键还原剂进行共同处理，经过诱导组装，分离，得到仿生纳米颗粒。
2.根据权利要求1所述的一种仿生纳米颗粒的制备方法，其特征在于：
所述含有二硫键和/或巯基的蛋白、多肽或多糖包括角蛋白、牛奶蛋白、酪蛋白、鱼皮蛋白、玉米蛋白、乳清蛋白、乳蛋白、蚕丝胶蛋白、乳球蛋白、豆浆蛋白、小麦蛋白、大豆蛋白、燕麦蛋白、胶原蛋白、乳铁蛋白、弹性蛋白、血清蛋白、麦醇溶蛋白、人纤维蛋白、胎盘蛋白、透明质酸、伸展蛋白、肌动蛋白、酵母蛋白、马铃薯蛋白、大米蛋白、米糠蛋白、木瓜蛋白酶、网硬蛋白、小麦谷蛋白、羽扇豆蛋白、芝麻蛋白、榛子蛋白、可溶性蛋白多糖或者其水解产物及修饰产物中的一种以上。
3.根据权利要求1所述的一种仿生纳米颗粒的制备方法，其特征在于：所述二硫键还原剂为半胱氨酸、二硫苏糖醇、还原型谷胱甘肽、β-巯基乙醇或三(2-羧乙基)膦。
4.根据权利要求1所述的一种仿生纳米颗粒的制备方法，其特征在于具体包括如下步骤：将含有二硫键和/或巯基的蛋白、多肽或多糖配制成质量分数0.1％～20％的溶液，加热升温至35～60℃，再加入浓度为0.1～200mM的二硫键还原剂水溶液，pH值调节至5.0～9.0，搅拌2～24小时，制备得到纳米颗粒溶液；再经过透析或离心，冻干，得到仿生纳米颗粒。
5.根据权利要求4所述的制备方法，其特征在于：所述透析是将纳米颗粒溶液放入透析袋，并于0～30℃的等渗缓冲溶液或纯水中透析除去多余的二硫键还原剂及其副产物；透析分子截留不低于1000；所述离心的转速为8000～15000rpm，10分钟/次，离心的次数为1～3次。
6.一种由权利要求1～5任一项所述的制备方法制备得到的仿生纳米颗粒，其特征在于：该仿生纳米颗粒由分子间巯基和/或二硫键重新装配成球状纳米结构。
7.根据权利要求6所述的仿生纳米颗粒在制备抗皮肤衰老产品、创面修复产品或药物递送产品中的用途。
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