CN117298131A - 一种用于糖尿病足的甘草酸-药根碱纳米药物、制备方法及其应用 - Google Patents
一种用于糖尿病足的甘草酸-药根碱纳米药物、制备方法及其应用 Download PDFInfo
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- CN117298131A CN117298131A CN202311608491.3A CN202311608491A CN117298131A CN 117298131 A CN117298131 A CN 117298131A CN 202311608491 A CN202311608491 A CN 202311608491A CN 117298131 A CN117298131 A CN 117298131A
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- Prior art keywords
- jateorhizine
- glycyrrhizic acid
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- solution
- drug
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Classifications
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- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
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- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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- C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
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- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
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Abstract
本发明涉及药物化学、药物制剂技术领域,特别涉及一种用于糖尿病足的甘草酸‑药根碱纳米药物、制备方法及其应用。本发明纳米药物由甘草酸与药根碱反应制备得到,同时利用制备的甘草酸‑药根碱纳米药物与氧化石墨烯‑银纳米复合材料相结合制备水凝胶敷料,实现对多重耐药金黄色葡萄球菌的耐药抑制作用,甘草酸‑药根碱纳米药物与氧化石墨烯‑银纳米复合材料联合使用呈现显著地协同增效抑菌作用,对多重耐药金黄色葡萄球菌的强大的抑菌效果,为糖尿病足的抗菌康复治疗提供了新的选择。
Description
技术领域
本发明涉及药物化学、药物制剂技术领域,特别涉及一种用于糖尿病足的甘草酸-药根碱纳米药物、制备方法及其应用。
背景技术
糖尿病患者感染风险大,皮肤、软组织和骨是糖尿病性感染的最常见部位,细菌感染以链球菌、金黄色葡萄球菌、革兰阴性杆菌及厌氧杆菌等最为多见;病毒感染则主要是带状疱疹和单纯疱疹;真菌感染以白色念珠菌为主。糖尿病足伤口的细菌常混合厌氧及嗜氧性的多种细菌,但常以厌氧性的细菌为主。感染时需耗掉更多的氧气去杀菌,因此会使组织缺氧的情形更为恶化;甚至一些接近正常边缘的血液循环(marginal blood flow)亦会受到严重之影响而死亡。细菌是一类个体微小、结构简单、生命力顽强的原核生物。它与人类的生活息息相关,其中有的是益生菌,大多数则是能够侵染人体诱发疾病的致病菌。根据革兰氏染色方法,可以将细菌划分为两类:革兰氏阳性菌和革兰氏阴性菌,革兰氏阳性菌的细胞壁厚20-80nm,主要由15-50层的肽聚糖和20-40%的磷壁酸组成。革兰氏阴性菌的细胞壁厚约10nm,由2-3层肽聚糖、脂多糖、单层磷脂外膜和脂多糖组成。革兰氏阴性菌和阳性菌的典型代表分别为大肠杆菌(Escherichiacoli, E.coli)和金黄色葡萄球菌(Staphylococcus aureus, S. aureus)。随着抗生素的滥用,越来越多的细菌获得了耐药性,甚至有的细菌获得了多重耐药性。耐药菌已经导致临床上出现用药困难的世界性难题,因此发展新型的抗菌剂已经迫在眉睫。
生物被膜的形成有助于保护MRSA免受宿主的免疫反应和抗生素制剂的影响,是其持续生存、氧化/环境压力和抗生素耐药性产生的关键因素之一。据统计,目前超过75%的细菌感染是由生物被膜介导的,这给全球医学界带来了治疗困难。鉴于这些挑战,开发具有高抗菌性能和生物被膜消除效果的抗菌剂及制定针对MRSA的多层面治疗策略势在必行。
纳米药物是指药剂学中的纳米粒或称纳米载体与纳米药物,其尺寸界定于1-1000nm之间。纳米载体是指溶解或分散有药物的各种纳米粒。纳米药物则是指直接将原料药加工成纳米粒。纳米给药系统的应用以及纳米材料作为抗菌剂的研究表明,纳米技术在细菌诱发的传染病治疗中具有巨大的潜力。纳米材料因其大的比表面积、良好的靶向性和良好的抑菌活性而受到广泛关注。例如,基于脂质的纳米结构被开发为有前途的纳米药物载体;银纳米颗粒优异的抗菌性能也受到广泛关注。然而,纳米载体载药量低以及金属纳米粒子对人体的潜在毒性等问题尚未得到解决。
发明内容
本发明为了弥补现有技术的不足,提供了一种用于糖尿病足的甘草酸-药根碱纳米药物、制备方法及其应用,主要用于糖尿病足创面护理,尤其是抗多重耐药菌金黄色葡萄球菌。
第一方面,本发明提供了一种用于糖尿病足的甘草酸-药根碱纳米药物,甘草酸-药根碱纳米药物由甘草酸与药根碱反应,采用分子截留制得,其结构如式(Ⅰ)所示:
式(Ⅰ)。
第二方面,本发明提供了甘草酸-药根碱纳米药物的制备方法,包括以下步骤:
(1)称取甘草酸溶解于热水中,加入氢氧化钾溶液调整溶液pH值为7.6-7.8,控制温度80℃,搅拌60min,得游离甘草酸溶液;
(2)称取药根碱溶解于热水中,加入磷酸溶液调整pH值为6.5-7.0,控制温度80℃,搅拌60min得药根碱溶液;
(3)将步骤(1)的游离甘草酸溶液缓慢加入步骤(2)的药根碱溶液中,控制温度90℃,搅拌120min;
(4)将步骤(3)的混合溶液置于透析袋内,采用超纯水透析18h以除去非反应的甘草酸、药根碱以及氢氧化钾、磷酸,每2h换一次超纯水,得到甘草酸-药根碱溶液;
(5)将步骤(4)制得的甘草酸-药根碱溶液离心,离心后,收集沉淀,超纯水洗涤,真空冷冻干燥,得到甘草酸-药根碱纳米药物。
进一步的,步骤(1)中氢氧化钾溶液的浓度为1mg/mL、3mg/mL、5mg/ml或10mg/mL中的一种。
进一步的,步骤(3)中甘草酸与药根碱的摩尔比为5:1、3:1、1:1、1:3或1:5中的一种。
进一步的,步骤(4)中透析袋的截留分子量为1.0 kDa、2.0 kDa或3.0 kDa中的一种。
进一步的,步骤(5)中离心转速为15000 rpm、20000 rpm或30000 rpm,离心时间为9min、10min、15 min或20min。
第三方面,本发明提供了甘草酸-药根碱纳米药物在制备水凝胶敷料中的应用。
第四方面,本发明提供了一种甘草酸-药根碱纳米药物在制备水凝胶敷料中的应用方法,包括以下步骤:
(1)采用Hummers方法制备氧化石墨烯,取氧化石墨烯水溶液和硝酸银溶液,依次置于水溶液中,加热煮沸,再加入柠檬酸三钠,煮沸1小时后冷却至室温,用超纯水离心洗涤多次,加入水溶液得到氧化石墨烯-银纳米溶液;
(2)取甘草酸-药根碱纳米药物,氧化石墨烯-银纳米溶液,加入水凝胶敷料中制备成水凝胶敷料。
进一步的,步骤(1)中,石墨烯水溶液为4mg/mL、2mL,硝酸银溶液为40mg/ml、4毫升,柠檬酸三钠160mg,水溶液为400mL;步骤(2)中,甘草酸-药根碱纳米药物为10mg,氧化石墨烯-银纳米溶液为10ml。
进一步的,步骤(2)中,所述水凝胶敷料为含有水溶性高分子颗粒如羧甲基纤维素、果胶、海藻酸钠与橡胶粘性物混合加工而成的敷料。
附图说明
下面结合附图对本发明作进一步的说明。
图1为本发明的甘草酸-药根碱纳米药物的扫描电镜图;
图2为本发明的甘草酸-药根碱纳米药物的抗菌活力结果图,其中a.未处理-金黄色葡萄球菌表面皿;b. GO-Ag纳米复合材料处理-金黄色葡萄球菌表面皿;c. 甘草酸-药根碱纳米药物+GO-Ag纳米复合材料处理-金黄色葡萄球菌表面皿;
图3为本发明的甘草酸-药根碱纳米药物对金黄色葡萄球菌细胞壁作用图;a. GO-Ag纳米复合材料处理-金黄色葡萄球菌细胞形态,b.甘草酸-药根碱纳米药物+GO-Ag纳米复合材料处理-金黄色葡萄球菌细胞形态。
具体实施方式
下面结合具体实施方式对本发明作进一步详细的说明,以帮助本领域的技术人员对本发明的发明构思、技术方案有更完整、准确和深入的理解,本发明的保护范围包括但不限于以下实施例。
实施例1 甘草酸-药根碱纳米药物的配比筛选
选取摩尔比5:1、3:1、1.5:1、1:1或1:3的甘草酸与药根碱,分五组参照以下方法制备不同的甘草酸-药根碱纳米药物,并测定不同甘草酸-药根碱纳米药物的水合粒径和多分散性指数作为评估该纳米药物均匀性和稳定性的指标,见表1。
制备方法:
(1)称取甘草酸溶解于热水中,加入浓度为10mg/mL氢氧化钾溶液调整pH值为7.6-7.8,控制温度80℃,搅拌60min,得游离甘草酸溶液;
(2)称取药根碱溶解于热水中,加入磷酸溶液调整pH值为6.5-7.0,控制温度80℃,搅拌60min得药根碱溶液;
(3)按照甘草酸与药根碱的不同摩尔比例(5:1、3:1、1.5:1、1:1或1:3),将步骤(1)的游离甘草酸溶液缓慢加入步骤(2)的药根碱溶液中,控制温度90℃,搅拌120min;
(4)将步骤(3)的混合溶液置于透析袋(MWCO=3.0 kDa)内,采用超纯水1000ml透析18h以除去非反应的甘草酸、药根碱以及氢氧化钾、磷酸,每2h换一次超纯水,得到甘草酸-药根碱溶液,所述透析袋的截留分子量为3.0 k Da;
(5)将步骤(4)制得的甘草酸-药根碱溶液离心,离心转速为30000 rpm,离心时间9min,收集沉淀,超纯水洗涤2次,真空冷冻干燥,得到甘草酸-药根碱纳米药物。
表1不同摩尔比甘草酸与药根碱合成纳米药物的平均粒径和PDI数值表
表1结果表明,甘草酸与药根碱的摩尔比影响了绿原酸-药根碱纳米药物的平均水合粒径和多分散性指数(polydispersity,PDI)。当甘草酸与药根碱比从5:1变化到1:3时,甘草酸与药根碱纳米药物的平均水合粒径及PDI分别在2256±118.4nm到286.8±3.5nm、0.58±0.26到0.25±0.03范围内变化。与其它比例相比,摩尔比为1.5:1时制备的甘草酸与药根碱纳米药物的平均水合粒径最小,为286.8±3.5nm,且粒径分布较窄,其PDI为0.25±0.03。PDI作为评估纳米药物均匀性和稳定性的指标之一,PDI等于或小于0.3表示纳米药物均匀分布,小粒径则可保证甘草酸与药根碱纳米药物稳定存在于血液中。因此,选择甘草酸与药根碱1.5:1的摩尔比作为制备甘草酸与药根碱纳米药物的最佳配比。
实施例2 甘草酸-药根碱纳米药物制备
甘草酸-药根碱纳米药物制备:
(1)称取甘草酸溶解于热水中,加入浓度为10mg/mL氢氧化钾溶液调整pH值为7.6-7.8,控制温度80℃,搅拌60min,得游离甘草酸溶液;
(2)称取药根碱溶解于热水中,加入磷酸溶液调整pH值为6.5-7.0,控制温度80℃,搅拌60min得药根碱溶液;
(3)按照甘草酸与药根碱的摩尔比1.5:1的比例,将步骤(1)的游离甘草酸溶液缓慢加入步骤(2)的药根碱溶液中,控制温度90℃,搅拌120min;
(4)将步骤(3)的混合溶液置于透析袋(MWCO=3.0 kDa)内,采用超纯水1000ml透析18h以除去非反应的甘草酸、药根碱以及氢氧化钾、磷酸,每2h换一次超纯水,得到甘草酸-药根碱溶液,所述透析袋的截留分子量为3.0 k Da;
(5)将步骤(4)制得的甘草酸-药根碱溶液离心,离心转速为30000 rpm,离心时间9min,收集沉淀,超纯水洗涤2次,真空冷冻干燥,得得到如图1所示呈球形的甘草酸-药根碱纳米药物,其结构式如式Ⅰ所示。
采用质谱法鉴定制备得到的甘草酸-药根碱纳米药物,质谱数据如下:
质荷比(m/z)主要有 1159.478、821.3975、647.4456、471.3423、453.3417、435.3569、338.4381、323.4531、294.4569 等分子离子峰,经解析1159.478为甘草酸-药根碱的分子离子峰,821.3975为甘草酸分子离子峰,338.0805为药根碱分子离子峰;323.4531为药根碱脱甲基碎片离子峰、294.4569为药根碱脱二甲基及一亚甲基碎片离子峰;647.4456、471.3423、453.3417、435.3569为甘草酸碎片离子峰。其中,甘草酸-药根碱纳米药物的分子离子峰的质荷比m/z= 1159.478(M-2峰),与甘草酸-药根碱复合物理论质荷比相符(m/z=1161.309),说明本方法制备得到了甘草酸-药根碱纳米药物。
实施例3 用于糖尿病足的甘草酸-药根碱纳米药物制备
甘草酸-药根碱纳米药物与氧化石墨烯-银纳米复合材料制备水凝胶敷料。
按照实施例2的制备方法,制备甘草酸-药根碱纳米药物,加入氧化石墨烯-银纳米复合材料中制备水凝胶敷料,具体如下:
(1)采用改进后的Hummers方法制备氧化石墨烯,取浓度为4mg/mL氧化石墨烯水溶液2mL和40mg/ml的硝酸银溶液4毫升,依次置于400mL水溶液中,加热煮沸,再加入柠檬酸三钠160mg,煮沸1小时后冷却至室温,用超纯水离心洗涤多次,加入水溶液得到氧化石墨烯-银纳米溶液;
(2)取甘草酸-药根碱纳米药物10mg,氧化石墨烯-银纳米溶液10ml,加入常规水凝胶敷料中制备成水凝胶敷料,所述常规水凝胶敷料为含有水溶性高分子颗粒如羧甲基纤维素、果胶、海藻酸钠与橡胶粘性物混合加工而成的敷料。
实验例4:甘草酸-药根碱纳米药物抗菌试验
实验过程:
(1)细菌的培养:本实验采用的金黄色葡萄球菌作为研究对象。首先配制液体培养基:称取10g的蛋白胨,10g的NaCl和5g的酵母粉,溶于1L纯化水中,高温灭菌,密封备用。然后在超净台中用洁净的接种环挑取一个单克隆菌落置于1mL的Luria-Bertani(LB)培养基中,在37℃恒温摇床220 rpm 震荡过夜。次日按1:100(V:V)将菌液重新接种到新鲜的LB培养基中,继续培养2-2.5小时,直到菌液在600nm 波长处的吸光度(OD600)达到0.5,此时的细菌处于生长旺盛的对数期。后面实验均采用对数期的菌液(OD600=0.5)进行研究。
(2)细菌活力检测:MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide)法是一种细菌或者细胞的活力检测方法18.19。此方法能够准确的测定出的细菌或者细胞活力。将对数期(OD600=0.5)的菌液按1:10(V:V)重新接种到含10ug/mL GO-Ag纳米复合材料和含10ug/mL甘草酸-药根碱纳米药物+10ug/mL GO-Ag纳米复合材料的LB培养基中,置于37℃恒温摇床,220 rpm震荡培养,2.5小时后,将菌液重新稀释一定倍数,每孔100μL依次加入到96孔板中,再每孔加入20μL MTT,37℃恒温培养箱避光培养4-6小时,待甲臜充分形成后,去上清每孔再加入120μL的DMSO置于摇床上150 rpm 震荡15分钟溶解,用多功能酶标仪检测570nm波长下的吸光值。所有实验均重复3次。
(3)细菌平板计数:细菌平板计数法是细菌研究中常用的检测细菌活菌数的一种有效方法。首先配制好固体培养基,准备好LB平板:称取10g的蛋白胨,10g的NaCl,5g的酵母粉和15g的琼脂糖,溶于1L纯化水中,高温灭菌,倾倒于无菌的培养皿中第冷却凝固,密封,置于4℃备用。取对数期(OD600=0.5)的金黄色葡萄球菌菌液按1:10(V:V) 接种到含10ug/mL GO-Ag纳米复合材料和含10ug/mL甘草酸-药根碱纳米药物+10ug/mL GO-Ag纳米复合材料的LB培养基中,置于37℃恒温摇床,220 rpm震荡培养2.5小时,收集菌液,混匀。取100μL菌液用1xPBS溶液(pH 7.2)进行梯度稀释。然后取100μL稀释菌液均匀涂布在LB平板上。在无菌操作台放置半小时后,倒置于37℃恒温培养箱中培养12-16小时。取出平板,进行平板菌落计数并且拍照记录。平板上菌落数乘以稀释倍数即得到每毫升菌液中的总菌落数。
(4)扫描电镜法观察细菌的形貌:葡萄球菌菌液按1:10(V:V)接种于浓度为30ug/mL GO-Ag的新鲜LB培养基中以及浓度为30ug/mL甘草酸-药根碱纳米药物+30ug/mL GO-Ag纳米复合材料的新鲜LB培养基中,37℃恒温摇床220 rpm震荡培养2.5小时。然后用1xPBS溶液(pH 7.2),3500r/min离心洗涤数次。4℃下,用2.5%戊二醛固定4小时,再用1xPBS溶液(pH7.2)3500r/min离心洗涤三次。接着进行乙醇梯度脱水(分别为30%、50%、70%、85%、90%乙醇各一次,100%的乙醇2次,每次15分钟)。取适量的菌液滴于洁净的硅片上,置于带盖的洁净小皿中,自然风干。再用离子溅射仪喷上一层金纳米颗粒,最后进行扫描电镜(Scanningelectron microscope,SEM)镜检。
(5)细菌生长曲线:细菌生长曲线是指以时间为横坐标,细菌增长数目的对数为纵坐标,绘制的一条曲线。通常用细菌在600nm波长处的吸光值(OD600)表示细菌生长曲线的纵坐标。微生物研究中常用细菌生长曲线来观察和分析细菌的生长情况。取对数期的金黄色葡萄细菌按1:10(V:V)重新接种到含30ug/mL GO-Ag纳米复合材料的液体LB培养基中孵育,同事接种到含30ug/mL甘草酸-药根碱纳米药物+30ug/mL GO-Ag纳米复合材料的新鲜LB培养基中孵育。间隔一定时间后取适量菌液,测定菌液的OD600值,连续测定11个小时。绘制一条细菌的生长曲线。
(6)实时观测细菌的分裂:用共聚焦显微镜观察细菌的分裂。
实验结果
(1)抗金黄色葡萄球菌的抗菌活性结果
通过MTT法对不同浓度(2.5、5、10 ug/mL) 甘草酸-药根碱纳米药物+10ug/mL GO-Ag纳米复合材料的抗菌效果进行了测试,结果表明,GO-Ag纳米复合材料有一定的抑菌作用,加入甘草酸-药根碱纳米药物后抑菌作用显著增强,随着甘草酸-药根碱纳米药物的浓度增加,细菌的活力在递减,当甘草酸-药根碱纳米药物的浓度达到10 ug/mL时,细菌的活力降低为0。为进一步验证甘草酸-药根碱纳米药物的抗菌效果,将细菌与甘草酸-药根碱纳米药物及GO-Ag纳米复合材料共同孵育2.5小时后,收集菌液,稀释一定倍数进行平板计数。如图2所示,经10ug/mL GO-Ag 纳米复合材料处理2.5小时后仅有少量的细菌存活,经10ug/mL甘草酸-药根碱纳米药物+10ug/mL GO-Ag 纳米复合材料处理2.5小时后无细菌存活。表明本发明制备的甘草酸-药根碱纳米药物具有很好的抗菌效果。
(2)对金黄色葡萄球菌生长分裂的影响结果:
金黄色葡萄球菌在含30ug/mL 甘草酸-药根碱纳米药物+GO-Ag纳米复合材料的LB培养基中孵育11小时,细菌OD600值未见明显增加,而对照组细菌的OD600值显著增加,表明经30ug/mL 甘草酸-药根碱纳米药物处理后,金黄色葡萄球菌的生长可能受到了抑制或者细菌已经死亡。随后,本发明通过共聚焦显微镜对金黄色葡萄球菌的生长分裂情况进行分析。通过共聚焦显微镜实时观测细菌在LB平板上的生长情况。本发明发现对照组金黄色葡萄球菌在0.5小时后即可观察到细菌增殖,1.5小时后细菌已经分裂了数代,由原来的十几个变成了几十个细菌;而实验组中,金黄色葡萄球菌在含30ug/mL GO-Ag纳米复合材料的LB平板上培养0.5小时后,未见细菌分裂,1.5小时后偶见个别细菌分裂;金黄色葡萄球菌在含30 ug/mL甘草酸-药根碱纳米药物的LB平板上培养1.5小时未见细菌分裂。
为验证金黄色葡萄球菌经甘草酸-药根碱纳米药物+GO-Ag 纳米复合材料处理后,细菌是否还保持活性,本发明将金黄色葡萄球菌先与含30ug/mL甘草酸-药根碱纳米药物30ug/mL GO-Ag纳米复合材料的LB培养基孵育2.5小时,再将细菌重新按1:100(V:V)接种到新鲜的LB液体培养基中,继续培养,获得金黄色葡萄球菌的生长曲线。结果,GO-Ag纳米复合材料处理后的金黄色葡萄球菌重新接种到新鲜LB培养基后,前4.5小时,几乎未见细菌的OD600值增长;在4.5小时到8小时,可观察到金黄色葡萄球菌的OD600值有轻微的增加,生长曲线呈上升的趋势。在8小时后细菌恢复旺盛增殖,细菌数目呈现指数增长。因此本发明推测金黄色葡萄球菌在与GO-Ag纳米复合材料孵育时,细菌虽然停止分裂,但GO-Ag纳米复合材料并未杀死细菌,其依旧保持一定的活力,而在GO-Ag纳米复合材料被去除后,通过一段时间的调整,金黄色葡萄球菌可以恢复正常的分裂。甘草酸-药根碱纳米药物+GO-Ag 纳米复合材料处理后的金黄色葡萄球菌重新接种到新鲜LB培养基后,前8小时,几乎未见细菌的OD600值增长,在8小时后继续观察8小时仍是几乎未见细菌的OD600值增长。表明金黄色葡萄球菌在与甘草酸-药根碱纳米药物孵育时,被灭活杀死。
(3)对金黄色葡萄球菌细胞壁/膜完整性的影响结果
金黄色葡萄球菌是典型的革兰氏阳性菌,相较于革兰氏阴性菌的细胞壁,其细胞壁较厚,主要成分是致密的肽聚糖和磷壁酸。细胞壁作为细菌的第一道防线,对抗菌机制研究具有重要价值。
为研究甘草酸-药根碱纳米药物是否会破坏金黄色葡萄球菌的细胞壁或膜,首先将金黄色葡萄球菌经过30 ug/mL甘草酸-药根碱纳米药物+GO-Ag纳米复合材料处理2.5小时后,然后通过酒精梯度脱水喷金,制备好实验组样品,通过 SEM观察细菌的形貌。同法也制备了对照组金黄色葡萄球菌样品进行 SEM分析其形貌。从扫描电镜观察结果显示,GO-Ag纳米复合材料处理的金黄色葡萄球菌的细胞壁或膜未见明显破损,细菌的形态仍保持着球形或近似球形,但可以清晰地发现金黄色葡萄球菌的胞外有一层物质(细胞分泌物)覆盖在细菌的表面。甘草酸-药根碱纳米药物+GO-Ag 纳米复合材料处理的金黄色葡萄球菌的细胞壁可见明显破损。
以上所述的仅是本发明的一些实施方式,对于本领域的普通技术人员来说,在不脱离本发明创造构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。
Claims (10)
1.一种用于糖尿病足的甘草酸-药根碱纳米药物,其特征在于:甘草酸-药根碱纳米药物由甘草酸与药根碱反应,采用分子截留制得,其结构如式(Ⅰ)所示:
式(Ⅰ)。
2.一种权利要求1所述的甘草酸-药根碱纳米药物的制备方法,其特征在于,包括以下步骤:
(1)称取甘草酸溶解于热水中,加入氢氧化钾溶液调整溶液pH值为7.6-7.8,控制温度80℃,搅拌60min,得游离甘草酸溶液;
(2)称取药根碱溶解于热水中,加入磷酸溶液调整pH值为6.5-7.0,控制温度80℃,搅拌60min得药根碱溶液;
(3)将步骤(1)的游离甘草酸溶液缓慢加入步骤(2)的药根碱溶液中,控制温度90℃,搅拌120min;
(4)将步骤(3)的混合溶液置于透析袋内,采用超纯水透析18h以除去非反应的甘草酸、药根碱以及氢氧化钾、磷酸,每2h换一次超纯水,得到甘草酸-药根碱溶液;
(5)将步骤(4)制得的甘草酸-药根碱溶液离心,离心后,收集沉淀,超纯水洗涤,真空冷冻干燥,得到甘草酸-药根碱纳米药物。
3.根据权利要求2所述的制备方法,其特征在于,步骤(1)中氢氧化钾溶液的浓度为1mg/mL、3mg/mL、5mg/ml或10mg/mL中的一种。
4.根据权利要求2所述的制备方法,其特征在于,步骤(3)中甘草酸与药根碱的摩尔比为5:1、3:1、1:1、1:3或1:5中的一种。
5.根据权利要求2所述的制备方法,其特征在于,步骤(4)中透析袋的截留分子量为1.0kDa、2.0 kDa或3.0 kDa中的一种。
6.根据权利要求2所述的制备方法,其特征在于,步骤(5)中离心转速为15000 rpm、20000 rpm或30000 rpm,离心时间为9min、10min、15 min或20min。
7.一种权利要求1所述的甘草酸-药根碱纳米药物在制备水凝胶敷料中的应用。
8.一种根据权利要求7所述的应用,其特征在于,应用方法包括以下步骤:
(1)采用Hummers方法制备氧化石墨烯,取氧化石墨烯水溶液和硝酸银溶液,依次置于水溶液中,加热煮沸,再加入柠檬酸三钠,煮沸1小时后冷却至室温,用超纯水离心洗涤多次,加入水溶液得到氧化石墨烯-银纳米溶液;
(2)取甘草酸-药根碱纳米药物,氧化石墨烯-银纳米溶液,加入水凝胶敷料中制备成水凝胶敷料。
9.根据权利要求8所述的应用,其特征在于,步骤(1)中,石墨烯水溶液为4mg/mL、2mL,硝酸银溶液为40mg/ml、4毫升,柠檬酸三钠160mg,水溶液为400mL;步骤(2)中,甘草酸-药根碱纳米药物为10mg,氧化石墨烯-银纳米溶液为10ml。
10.根据权利要求8所述的应用,其特征在于,步骤(2)中,所述水凝胶敷料为含有水溶性高分子颗粒如羧甲基纤维素、果胶、海藻酸钠与橡胶粘性物混合加工而成的敷料。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010051683A1 (zh) * | 2008-11-07 | 2010-05-14 | 北京宏泰康达医药科技有限公司 | 一种治疗急性肠炎和痢疾的组合物 |
CN102107006A (zh) * | 2009-12-25 | 2011-06-29 | 奇复康药物研发(苏州)有限公司 | 用于治疗糖尿病的偶联药物及其医药用途 |
CN102357079A (zh) * | 2011-10-28 | 2012-02-22 | 复旦大学 | 一种甘草酸修饰羧甲基壳聚糖纳米粒及其制备方法和应用 |
CN103250739A (zh) * | 2012-08-20 | 2013-08-21 | 苏州大学 | 氧化石墨烯/银颗粒纳米复合物的制备方法及应用 |
CN114796272A (zh) * | 2022-04-13 | 2022-07-29 | 中国科学院苏州生物医学工程技术研究所 | 银纳米@碳点复合协同抗菌材料、其应用及抗菌药物 |
CN115252560A (zh) * | 2022-07-28 | 2022-11-01 | 中国药科大学 | 一种基于天然产物的自组装纳米粒及其制备方法和应用 |
CN116617220A (zh) * | 2023-07-26 | 2023-08-22 | 南京农业大学 | 抗青霉素类耐药菌的绿原酸-小檗碱纳米药物、药物组合物及其制备方法 |
-
2023
- 2023-11-29 CN CN202311608491.3A patent/CN117298131B/zh active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010051683A1 (zh) * | 2008-11-07 | 2010-05-14 | 北京宏泰康达医药科技有限公司 | 一种治疗急性肠炎和痢疾的组合物 |
CN102107006A (zh) * | 2009-12-25 | 2011-06-29 | 奇复康药物研发(苏州)有限公司 | 用于治疗糖尿病的偶联药物及其医药用途 |
CN102357079A (zh) * | 2011-10-28 | 2012-02-22 | 复旦大学 | 一种甘草酸修饰羧甲基壳聚糖纳米粒及其制备方法和应用 |
CN103250739A (zh) * | 2012-08-20 | 2013-08-21 | 苏州大学 | 氧化石墨烯/银颗粒纳米复合物的制备方法及应用 |
CN114796272A (zh) * | 2022-04-13 | 2022-07-29 | 中国科学院苏州生物医学工程技术研究所 | 银纳米@碳点复合协同抗菌材料、其应用及抗菌药物 |
CN115252560A (zh) * | 2022-07-28 | 2022-11-01 | 中国药科大学 | 一种基于天然产物的自组装纳米粒及其制备方法和应用 |
CN116617220A (zh) * | 2023-07-26 | 2023-08-22 | 南京农业大学 | 抗青霉素类耐药菌的绿原酸-小檗碱纳米药物、药物组合物及其制备方法 |
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