CN114558144B - 一种新型黑木耳多糖包裹碳纳米管负载阿霉素材料的制备及其应用 - Google Patents
一种新型黑木耳多糖包裹碳纳米管负载阿霉素材料的制备及其应用 Download PDFInfo
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Abstract
本发明公开了一种新型黑木耳多糖包裹碳纳米管负载阿霉素材料的制备及其应用，属于纳米材料领域。通过将黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液混合后搅拌反应得到本发明材料，黑木耳多糖包裹碳纳米管负载阿霉素显著提高了碳管和疏水性阿霉素的分散性和稳定性，该材料具有极好的光热和pH响应性缓释效果，极大的减轻了正常细胞的毒副作用提高了抗肿瘤疗效。本发明解决了传统化疗药物效果差、半衰期短、毒性大，以及用药量高的问题，对抗肿瘤新药的研发以及新的抗肿瘤结合手段的发展具有重要意义。
Description
技术领域
本发明属于纳米材料领域，具体涉及一种黑木耳多糖包裹碳纳米管负载阿霉素材料的制备及其抗肿瘤应用。
背景技术
纳米药物因其独特的特性，在肿瘤的药物传递、成像、诊断和治疗等方面日益受到人们的关注。纳米结构负载的药物被认为具有比化疗药物更好的治疗指标，增强了溶解度、稳定性、半稳态、药代动力学和靶部位的积累[1；2]。许多研究报道纳米技术可以促进一些生物分子的传递，包括DNA、mRNA和蛋白质到它们的作用位点[3；4]。而且，与传统治疗相比，它们可以达到“正确的靶点”和“正确的照射”肿瘤部位，最终减少不必要的毒性，提高治疗指标[5；6；7]。然而，传统的给药方式仍然面临许多挑战，如对三阴性乳腺癌的特异性差、药物进入转移部位的效率低、细胞和肿瘤水平上的耐药性、药物不理想的理化特性以及难以消除肿瘤干细胞。因此，需要设计一种新型纳米载体系统，满足以下条件：(1)靶向释放肿瘤部位；(2)对正常细胞的细胞毒性低；(3)提高药物的半衰期；(4)根除肿瘤干细胞的有效方法。
发明内容
针对现有技术存在的上述缺陷，本发明的目的在于提供一种新型黑木耳多糖(BFP)包裹碳纳米管负载阿霉素材料(BSD)的制备及其在制备抗肿瘤药物中的应用。本发明通过黑木耳多糖包裹碳纳米管负载阿霉素，显著提高了碳管和疏水性阿霉素的分散性和稳定性。
本发明的目的通过下述技术方案实现：
一种新型黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，包括以下步骤：
(1)以DMSO为溶剂，分别配制黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液。
(2)在避光条件下，黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液混合均匀后充分反应，得到黑木耳多糖包裹碳纳米管负载阿霉素材料。
步骤(1)中，所述的经酸氧化改性的碳纳米管溶液通过将经酸氧化改性的碳纳米管超声分散在DMSO中得到。所述的经酸氧化改性的碳纳米管优选通过包括以下步骤的方法得到：将经酸氧化改性的碳纳米管超声分散在DMSO中得到。所述的经酸氧化改性的碳纳米管优选通过包括以下步骤的方法得到：将碳纳米管加到由浓硫酸和浓HNO3组成的混酸中，并在冷凝回流管中冷凝回流2-6h。反应液过滤后用水洗涤，收集黑色固体后冷冻干燥得到氧化改性的碳纳米管。所述的混酸优选由浓硫酸(质量分数为98.3％)和浓HNO3(质量分数为68％) 按体积比体积比为3:1组成。
步骤(2)中，混合溶液中黑木耳多糖、经酸氧化改性的碳纳米管、阿霉素的质量比优选为5:1:10-20:1:10。
进一步地，步骤(2)为：在避光条件下，将黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液混合后超声10-30min使碳纳米管充分分散并且与其他溶液混合均匀，再搅拌12h-24h使其充分反应，得到黑木耳多糖包裹碳纳米管负载阿霉素材料。
进一步地，步骤(2)还包括透析、冻干的步骤：反应产物在透析袋中用水透析除去未反应的原料，最后冻干得到黑木耳多糖包裹碳纳米管负载阿霉素材料。所述的透析袋优选为分子量截留为3500Da的透析袋。
上述黑木耳多糖为提取自黑木耳的具有三螺旋结构的β-1,3-D-葡聚糖，其主链为带有三个葡萄糖残基，侧链为带有两个葡萄糖残基的重复单元结构多糖。
上述碳纳米管可为多壁碳纳米管，也可为单壁碳纳米管。
一种新型黑木耳多糖包裹碳纳米管负载阿霉素材料，通过上述制备方法得到。该材料在近红外激光区660nm波长下具有极好的近红外效应，同时抗肿瘤药物阿霉素的负载，赋予其具有协同光热治疗和化疗相结合的抗肿瘤功能。在体外模拟的正常生理条件(pH7.4)和肿瘤条件(pH 5.0)下，该材料具有在正常生理条件下释放缓慢，在肿瘤条件下释放速率增快，且累计释放量在肿瘤条件下几乎是正常条件下的3倍，具有pH响应释放特点，可以靶向肿瘤释放。
利用所述的新型黑木耳多糖包裹碳纳米管负载阿霉素材料进行化疗与光热联合治疗，可通过促进肿瘤细胞内氧化应激，扰乱代谢和能量途径，达到清除原发肿瘤和抑制肿瘤转移的目的。所述的新型黑木耳多糖包裹碳纳米管负载阿霉素材料在杀死转移性三阴性乳腺癌方面表现出卓越的性能。
所述的新型黑木耳多糖包裹碳纳米管负载阿霉素材料具有制备抗肿瘤药物的应用。
一种抗肿瘤药物，包含所述的新型黑木耳多糖包裹碳纳米管负载阿霉素材料。
本发明的优点和有益效果：
本发明利用天然黑木耳多糖来提高碳管的分散性和阿霉素的水溶性，同时结合了碳管具备的光热效果，以及阿霉素的pH响应释放性，制备出了一种结合光热-缓释响应性化疗一体化的肿瘤治疗药物。
本发明成品为棉絮状固体，易于储存和运输。
本发明材料效果明显，通过体外细胞实验表明，黑木耳多糖包裹碳纳米管负载阿霉素材料对乳腺癌细胞具有极好的杀伤效果，但对正常细胞的毒性大大降低，符合缓释药物标准。
本发明提供了一种创新的杀伤乳腺癌的纳米药物，该药物具有极好的光热和化疗结合特点，是一种具有应用前景的抗肿瘤纳米药物。
本发明解决了传统化疗药物效果差、半衰期短、毒性大，以及用药量高的问题，对抗肿瘤新药的研发以及新的抗肿瘤结合手段的发展具有重要意义。
附图说明
图1为本发明中黑木耳(a)、BFP(b，c)、BSD水溶液(d)的样品图，其中c为BFP 溶解于水溶液中；
图2为本发明中BSD的扫描电镜图(a)和透射电镜图(b)；
图3为本发明中不同浓度BSD在近红外激光660nm下的光热效果图和光热循环稳定性图；
图4为本发明中BSD在不同pH下的药物累积释放(a)和释放速率图(b)；
图5为本发明中BSD和纯DOX的体内摄取效果图；
图6为本发明中BSD对乳腺癌细胞(a)和正常细胞(b)的细胞活性图。
具体实施方式
下面结合实施例对本发明作详细说明，但本发明的保护范围不限于下述的实施例。
实施例1黑木耳多糖包裹碳纳米管负载阿霉素材料(BSD)的制备
(1)BFP-DMSO溶液的配制：黑木耳多糖通过水热提取法获得[8]，原材料黑木耳(图1a)产地为福建厦门。具体步骤为：将250g黑木耳搅碎后用丙酮和乙酸乙酯分别回流4h脱脂，接着用在室温下将其浸入70％乙醇/水溶液中24h以去除上清液，残留物用0.15M NaCl水溶液在80-100℃下萃取2h，然后将溶液搅拌过夜。这个过程重复了三遍。将所得溶液离心得到上清液，再在25℃下用70％乙醇/水沉淀进一步纯化透析干燥得到黑木耳多糖BFP。将一定量浓度多糖BFP(1-5mg/mL，图1b)完全溶解于DMSO中，并持续搅拌2-24h，得到澄清透亮的多糖溶液(图1c)。
(2)氧化改性的碳纳米管-DMSO溶液的配制：将10g单壁碳纳米管加入300mL由浓硫酸(质量分数为98.3％)和浓HNO3(质量分数为68％)按体积比1:3组成混酸中搅拌均匀，接着在冷凝回流管中于100℃冷凝回流3h。接着用超纯水过滤洗涤5次，收集黑色固体后冷冻干燥获得氧化改性的碳纳米管。称取2mg氧化改性的碳纳米管将其超声分散在2mL DMSO中。
(3)DOX-DMSO溶液的配制：将20mg盐酸阿霉素溶于1mL DMSO中，加入20μL三乙胺持续搅拌反应12h脱盐酸得到20mg/mL的阿霉素溶液。或者直接使用阿霉素溶于DMSO 中，保持浓度为20mg/mL。
(4)将20mL 1mg/mL BFP-DMSO溶液、2mL 1mg/mL氧化改性的碳纳米管-DMSO溶液和1mL 20mg/mL的DOX-DMSO溶液混合超声10min，再搅拌12h，过程一直避光处理。随后，将反应产物在分子量截留为3500Da的透析袋中用纯水透析3天，除掉未反应的小分子原料，最后冻干得到黑木耳多糖包裹碳纳米管负载阿霉素材料(BSD)。
图1d中显示出本实施例1中得到的BSD溶于水中得到的分散液具有极好的分散性和稳定性。
图2为本实施例1所得BSD的扫描电镜和透射电镜图，表明多糖缠绕在碳管上，说明碳管可能通过多糖的缠绕来大大提高其分散性和稳定性。
图3为本实施例1所得BSD在不同浓度水溶液下的近红外光热成像图，以及其经过多次激光开-关循环温度图，BSD水溶液的浓度从0-100μg/mL，其光热升高温度变化从5-45℃改变，表明BSD具有极好的光热效应，除此之外，经过7次的光热循环实验，发现其光热循环稳定性也极好。
实施例2BSD药物装载能力
DOX负载能力(DLC)定义为DOX在BSD中的重量百分比，DOX封装效率(DEE) 定义为装载的DOX在BSD中的重量百分比和初始DOX的重量百分比。首先称取0、5、10、 20μg/mL DOX水溶液在495nm下测得一元线性回归方程，接着将实施例1中反应混合的 BSD-DMSO溶液在100mL水溶液中进行透析，每天透析三次，连续透析3天后测得每次溶液中的DOX质量，最后用初始加入的DOX质量减去透析掉DOX质量，最终得到装载的DOX 质量。将最终获得的BSD冻干透析称重，即得BSD的总质量，最终测得BSD的DLC和DEE 分别为43.5％和89.3％。
将30mL 1mg/mL BSD装入透析袋(Mw＝3500)中，置于37℃的磷酸盐缓冲液(pH7.4) 和乙酸缓冲液(pH 5.0)摇瓶中连续振荡，模拟BSD体内药物缓释。在特定时间点收集5mL 透析液并补加相同体积的缓冲液，确保透析液总体积不变。通过紫外-可见分光光度计测定透析液吸光度。
图4为本实施例1所得BSD在不同pH下的体外模拟释放图，结果表明在正常生理条件下(pH 7.4)BSD中的DOX释放速度极慢；而在pH 5.0的缓冲溶液中达到两相释放：在最初的24h，BSD达到极快的释放，累积释放达约15％，随后DOX的释放变慢，最终在16天后累积释放达30％。说明BSD具有极好的pH响应释放性，能够靶向具有酸性pH的肿瘤细胞。
实施例3DOX和BSD中的DOX在细胞体内摄取
1×104乳腺癌MDA-MB-231细胞接种于24孔板培养皿中，置于0.5mL新鲜DMEM完全培养基中培养12h，处理组和对照组各3个重复。将含5μg/mL BSD(BSD中DOX质量分数为43.5％)和2μg/mL DOX的培养基加入进乳腺癌细胞中接着分别培养4h、10h和24h。用 2.5μL/孔的1mg/mL 4',6-二脒基-2-苯基吲哚(DAPI，一种细胞核染色剂，会将核染为蓝色，购自碧云天生物有限公司)染色细胞核20min，用荧光倒置显微镜记录BSD和游离DOX的细胞摄取图像。
图5为本实施例1所得BSD和纯DOX在乳腺癌细胞体内的摄取图，结果表明纯的DOX在4h就被细胞摄取进细胞核，而BSD在4h只是被摄取进细胞质，直到24h才完全进入细胞核，说明BSD具有极好的缓释效果。
实施例4细胞活性测试
2×104细胞/孔接种于96孔板培养24h，将不同浓度的BSD加入乳腺癌细胞MDA-MB-231 中，共培养24h。同时用不同浓度的BSD处理正常细胞L02 24h。采用MTT试剂盒检测经过处理24h后的细胞活力，采用SPARK 10M酶标仪测细胞处理后在490nm处吸光度。以处理组和对照组的吸光度比值来计算细胞活力，结果以百分数表示。
图6为本实施例1所得BSD对乳腺癌细胞MDA-MB-231和正常细胞L02的细胞活性测试，结果表明BSD对乳腺癌细胞具有BSD对乳腺癌细胞具有极好的杀伤效果，但对正常细胞的毒性大大降低，说明BSD是一种极有潜力的抗肿瘤药物。
以上所述仅为本发明的示例性实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
参考文献：
[1]Y.Jin,L.Cai,Q.Yang,Z.Luo,L.Liang,Y.Liang,B.Wu,L.Ding,D.Zhang,X.Xu,L.Zhang,and F.Zhou,Anti-leukemia activities of selenium nanoparticlesembedded in nanotube consisted of triple-helixβ-d-glucan.Carbohyd.Polym.240(2020)116329.
[2]M.Li,Z.Tang,S.Lv,W.Song,H.Hong,X.Jing,Y.Zhang,and X.Chen,Cisplatincrosslinked pH-sensitive nanoparticles for efficient delivery ofdoxorubicin.Biomaterials 35(2014) 3851-3864.
[3]W.Zhao,H.Qian,R.Zhang,X.Gao,and X.Gou,MicroRNA targetingmicrotubule cross-linked protein(MACF1)would suppress the invasion andmetastasis of malignant tumor.Med. Hypotheses 104(2017)25-29.
[4]X.Piao,H.Yin,S.Guo,H.Wang,and P.Guo,RNA Nanotechnology toSolubilize Hydrophobic Antitumor Drug for Targeted Delivery.Advanced Science6(2019)1900951.
[5]Z.Ping,T.Liu,H.Xu,Y.Meng,W.Li,X.Xu,and L.Zhang,Construction ofhighly stable selenium nanoparticles embedded in hollow nanofibers ofpolysaccharide and their antitumor activities.Nano Res.10(2017)3775-3789.
[6]M.Li,Z.Tang,D.Zhang,H.Sun,H.Liu,Y.Zhang,Y.Zhang,and X.Chen,Doxorubicin-loaded polysaccharide nanoparticles suppress the growth of murinecolorectal carcinoma and inhibit the metastasis of murine mammary carcinomain rodent models.Biomaterials51(2015)161-172.
[7]S.Ashton,Y.H.Song,J.Nolan,E.Cadogan,J.Murray,R.Odedra,J.Foster,P.A.Hall,S.Low,P.Taylor,R.Ellston,U.M.Polanska,J.Wilson,C.Howes,A.Smith,R.J.A.Goodwin,J.G.Swales, N.Strittmatter,Z.Takáts,A.Nilsson,P.Andren,D.Trueman,M.Walker,C.L.Reimer,G.Troiano,D.Parsons,D.De Witt,M.Ashford,J.Hrkach,S.Zale,P.J.Jewsbury,and S.T.Barry,Aurora kinase inhibitornanoparticles target tumors with favorable therapeutic index invivo.Sci.Transl.Med.8(2016)317r-325r.
[8]S.Xu,X.Xu,and L.Zhang,Branching Structure and Chain Conformationof Water-Soluble Glucan Extracted from Auricularia auricula-judae.J.Agr.FoodChem.60(2012)3498-3506。
Claims (10)
1.一种黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：包括以下步骤：
（1）以DMSO为溶剂，分别配制黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液；所述黑木耳多糖为提取自黑木耳的具有三螺旋结构的β-1,3-D-葡聚糖，其主链为带有三个葡萄糖残基，侧链为带有两个葡萄糖残基的重复单元结构多糖；
（2）在避光条件下，黑木耳多糖溶液、经酸氧化改性的碳纳米管溶液、阿霉素溶液混合均匀后充分反应，反应产物在透析袋中用水透析除去未反应的原料，冻干得到黑木耳多糖包裹碳纳米管负载阿霉素材料。
2.根据权利要求1所述的黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：步骤（1）中，所述的经酸氧化改性的碳纳米管溶液通过将经酸氧化改性的碳纳米管超声分散在DMSO中得到。
3.根据权利要求1所述的黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：步骤（1）中，所述的经酸氧化改性的碳纳米管通过包括以下步骤的方法得到：将碳纳米管加到由浓硫酸和浓HNO3组成的混酸中，并在冷凝回流管中冷凝回流2-6h；反应液过滤后用水洗涤，收集黑色固体后冷冻干燥得到氧化改性的碳纳米管。
4.根据权利要求3所述的黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：所述的混酸由浓硫酸和浓HNO3按体积比体积比为1:3组成。
5.根据权利要求1所述的黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：所述的碳纳米管为多壁碳纳米管或单壁碳纳米管。
6.根据权利要求1所述的黑木耳多糖包裹碳纳米管负载阿霉素材料的制备方法，其特征在于：步骤（2）中，混合溶液中黑木耳多糖、经酸氧化改性的碳纳米管、阿霉素的质量比为5:1:10-20:1:10。
7.一种黑木耳多糖包裹碳纳米管负载阿霉素材料，其特征在于：通过权利要求1-6任一项所述的制备方法得到。
8.权利要求7所述的黑木耳多糖包裹碳纳米管负载阿霉素材料在制备抗肿瘤药物中的应用。
9.一种抗肿瘤药物，其特征在于：包含权利要求7所述的黑木耳多糖包裹碳纳米管负载阿霉素材料。
10.根据权利要求8所述的应用或权利要求9所述的抗肿瘤药物，其特征在于：所述的肿瘤包括乳腺癌。
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