CN116617220B - 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 - Google Patents
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 Download PDFInfo
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Abstract
本发明公开了一种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,该纳米药物由绿原酸与小檗碱采用透析沉淀法自组装制得。本发明利用制备的绿原酸‑小檗碱无载体纳米药物对青霉素类耐药菌的erm、mecA及/或pbpB耐药基因以及生物被膜基因表达进行抑制,实现对多重耐药金黄色葡萄球菌的耐药抑制作用,并将绿原酸‑小檗碱无载体纳米药物与氨苄青霉素组合作为药物组合物,使二者呈现显著地协同抑菌作用,从而使氨苄青霉素重新表现出对多重耐药金黄色葡萄球菌的强大的抑菌效果,具有良好的缓释性和生物安全性,为耐药细菌感染治疗提供了新的选择。
Description
技术领域
本发明涉及药物化学技术领域,更具体地,本发明涉及一种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,应用于抗青霉素类耐药菌,尤其是抗多重耐药金黄色葡萄球菌。
背景技术
在过去的几十年里,细菌的抗生素耐药已经对人类健康和畜牧养殖业的发展造成了严重威胁。尤其耐多药金黄色葡萄球菌(multidrug-resistant Staphylococcusaureus,MRSA)对目前主流的大多数β-内酰胺类抗生素如青霉素、头孢菌素和四环素等已产生耐药,导致如青霉素、头孢菌素和四环素等大多数β-内酰胺类抗生素无法对多重耐药黄金色葡萄球菌造成的感染进行治疗。
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。因此,寻找一种由植物药效成分直接自组装,不借助任何载体递药的纳米药物递送系统显得尤为重要。
发明内容
本发明的目的在于提供了一种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。
根据本发明的一个方面,提供了抗青霉素类耐药菌的绿原酸-小檗碱纳米药物,纳米药物由绿原酸与小檗碱采用透析沉淀法自组装制得,其结构如式(Ⅰ)所示:
式(Ⅰ)。
根据本发明的再一个方面,制备抗青霉素类耐药菌的绿原酸-小檗碱纳米药物的方法,包括以下步骤:
(1)称取绿原酸与小檗碱分别溶解于甲醇中,使用氢氧化钠溶液调整绿原酸的甲醇溶液的pH值为7.0-7.5,将上述两种溶液搅拌混匀,缓慢加入至磷酸缓冲液中恒温搅拌,便于绿原酸与小檗碱进行自组装,得到混合溶液;
(2)将步骤(1)的混合溶液置于透析袋内,采用超纯水透析24h以除去非自组装的小檗碱、绿原酸以及有机溶剂,每4h换一次超纯水,得到绿原酸与小檗碱自组装溶液;
(3)将步骤(2)制得的绿原酸与小檗碱自组装溶液离心,收集沉淀,超纯水洗涤3次,真空冷冻干燥,得到绿原酸-小檗碱无载体纳米药物。
在一些实施方式中,步骤(1)中小檗碱与绿原酸的摩尔比为1:4、1:2、1:1、2:1或4:1中的一种,氢氧化钠溶液的浓度为5mg/mL,磷酸缓冲溶液为60℃的磷酸缓冲溶液。
在一些实施方式中,步骤(2)中透析袋的截留分子量为3.5 kDa,步骤(3)中离心转速为12000 rpm,离心时间为10-20 min。
根据本发明的第三个方面,提供了抗青霉素类耐药菌的药物组合物,包括青霉素类抗生素和抗青霉素类耐药菌的绿原酸-小檗碱纳米药物。
在一些实施方式中,青霉素类抗生素为氨苄青霉素,绿原酸-小檗碱纳米药物的自组装结构由静电引力、π-π堆叠作用和氢键作用维持。
在一些实施方式中,氨苄青霉素与绿原酸-小檗碱纳米药物复配的摩尔质量比为:190~210:2~4。
在一些实施方式中,氨苄青霉素与绿原酸-小檗碱纳米药物复配的摩尔比为:200:3,绿原酸-小檗碱纳米药物的浓度为该纳米药物对多重耐药金黄色葡萄球菌的最小抑菌浓度值的二分之一。
根据本发明的第四个方面,提供了绿原酸-小檗碱纳米药物及/或抗青霉素类耐药菌的药物组合物在制备抗青霉素类耐药菌杀菌剂中的应用,抗青霉素类耐药菌为携带青霉素类耐药基因erm、mecA及/或pbpB基因的葡萄球菌科细菌。
在一些实施方式中,抗青霉素类耐药菌为多重耐药金黄色葡萄球菌。
本发明的有益效果:本发明公开了一种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,利用制备的绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌(多重耐药金黄色葡萄球菌)的耐药基因erm、mecA及/或pbpB基因表达抑制,实现对多重耐药金黄色葡萄球菌的耐药抑制作用,绿原酸-小檗碱无载体纳米药物与氨苄青霉素联合使用呈现显著地协同增效抑菌作用,从而使氨苄青霉素重新表现出对多重耐药金黄色葡萄球菌的强大的抑菌效果,具有良好的缓释性和生物安全性,能够有效杀灭耐药菌,为耐药细菌感染治疗提供了新的选择。
附图说明
图1为本发明的绿原酸-小檗碱无载体纳米药物的透射电镜图。
图2为本发明的绿原酸-小檗碱无载体纳米药物的质谱图。
图3为本发明的绿原酸-小檗碱无载体纳米药物的细胞毒性分析结果图。
图4为本发明的不同浓度的绿原酸、小檗碱、绿原酸-小檗碱无载体纳米药物、苯唑西林钠及氨苄青霉素的抑菌结果图,其中A为不同浓度各种药物对金黄色葡萄球菌的抑制率,B为不同浓度各种药物对多重耐药金黄色葡萄球菌的抑制率。
图5为本发明的抗青霉素类耐药菌的药物组合物对多重耐药金黄色葡萄球菌的抑菌结果图。
图6为本发明的抗青霉素类耐药菌的药物组合物对青霉素类耐药菌的耐药基因表达影响结果图。
图7为本发明的绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的生物被膜相关基因的影响。
图8为本发明的绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的营养吸收及核苷酸代谢相关基因表达的影响。
具体实施方式
通过具体的实施案例对本发明作进一步详细的描述,应理解这些实施例仅用于说明本发明而不用于限制本发明的保护范围,在阅读了本发明之后,本领域技术人员对本发明的各种等价形式的修改均落于本申请所附权利要求所限定。若无特殊说明,本发明的所有原料和试剂均为常规市场可购买的原料、试剂。
实施例1 绿原酸-小檗碱无载体纳米药物的配比筛选
以小檗碱与绿原酸的摩尔比1:4、1:2、1:1、2:1、4:1,为五个处理,参照以下方法制备不同的绿原酸-小檗碱无载体纳米药物,并测定不同绿原酸-小檗碱无载体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)称取称取不同摩尔比的小檗碱与绿原酸分别溶解于甲醇中,用5mg/mL的氢氧化钠溶液调整绿原酸的甲醇溶液的pH 值为7.0-7.5,随后,将小檗碱的甲醇溶液与绿原酸的甲醇溶液搅拌混匀,缓慢加入至10 mL 60℃的磷酸缓冲液中恒温搅拌30min,便于绿原酸与小檗碱进行自组装,得到混合溶液;
(2)将步骤(1)的混合溶液置于透析袋(MWCO=3.5 kDa)内,采用500 mL超纯水透析24 h,以除去非自组装的小檗碱、绿原酸以及有机溶剂,每4h换一次超纯水,得到绿原酸与小檗碱自组装溶液;
(3)将步骤(2)制得的绿原酸与小檗碱自组装溶液以12000 rpm的转速离心10-20min,收集沉淀,超纯水洗涤3次,真空冷冻干燥,得到绿原酸-小檗碱无载体纳米药物。
表1不同摩尔比小檗碱与绿原酸合成纳米药物的平均粒径和PDI数值表
如表1所示,小檗碱(BBR)与绿原酸(CGA)的摩尔比影响了绿原酸-小檗碱无载体纳米药物的平均水合粒径和多分散性指数(polydispersity,PDI)。当小檗碱与绿原酸的摩尔比从1:4变化到4:1时,绿原酸-小檗碱无载体纳米药物的平均水合粒径及PDI分别在1936.7±123.9到358.6±2.4 nm、1.00±0.00到0.28±0.01范围内变化。与其它比例相比,摩尔比为1:1时制备的绿原酸-小檗碱无载体纳米药物的平均水合粒径最小,为358.6±2.4 nm,且粒径分布较窄,其PDI为0.28±0.01。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,小粒径则可保证绿原酸-小檗碱无载体纳米药物稳定存在于血液中。因此,选择小檗碱与绿原酸1:1的摩尔比作为制备绿原酸-小檗碱无载体纳米药物的最佳配比。
实施例2 抗青霉素类耐药菌的药物组合物的制备
2.1 绿原酸-小檗碱无载体纳米药物的制备
(1)称取摩尔比为1:1的小檗碱与绿原酸分别溶解于甲醇中,用5 mg/mL的氢氧化钠溶液调整绿原酸的甲醇溶液的pH 值为7.0-7.5,随后,将小檗碱的甲醇溶液与绿原酸的甲醇溶液搅拌混匀,缓慢加入至10 mL 60℃的磷酸缓冲液中恒温搅拌30 min,便于绿原酸与小檗碱进行自组装,得到混合溶液;
(2)将步骤(1)的混合溶液置于透析袋(MWCO=3.5 kDa)内,采用500 mL超纯水透析24 h,以除去非自组装的小檗碱、绿原酸以及有机溶剂,每4h换一次超纯水,得到绿原酸与小檗碱自组装溶液;
(3)将步骤(2)制得的绿原酸与小檗碱自组装溶液以12000 rpm的转速离心10-20min,收集沉淀,超纯水洗涤3次,真空冷冻干燥,得到如图1所示呈球形的绿原酸-小檗碱无载体纳米药物,其结构式如式Ⅰ所示。
采用质谱法鉴定制备得到的绿原酸-小檗碱无载体纳米药物,所得到的质谱图见图2。从质谱图可以看出,绿原酸-小檗碱无载体纳米药物的分子离子峰的质荷比m/z=688.2032,与绿原酸和小檗碱摩尔比1:1时自组装单元的质荷比相符(m/z=690.68),说明本方法制备得到了绿原酸-小檗碱无载体纳米药物。
2.2 抗青霉素类耐药菌的药物组合物的制备
取浓度为50 µM的氨苄青霉素溶液1 L,取0.75 µmol绿原酸-小檗碱无载体纳米药物加入氨苄青霉素溶液中,混合均匀,即制得抗青霉素类耐药菌的药物组合物。
实施例3 绿原酸-小檗碱无载体纳米药物的细胞毒性分析
利用实施例2制备得到的绿原酸-小檗碱无载体纳米药物进行细胞毒性分析,方法如下:
将小鼠乳腺上皮细胞按照每孔5000个细胞均匀传至96孔板,待细胞形态良好且培养至80%数量左右,进行细胞毒性试验。将绿原酸-小檗碱无载体纳米药物配置成终浓度为0、0.75、1.5、3、6和12 μM的工作液,取100 µL各浓度的工作液加入96孔板中,培养24 h后弃掉培养液,用无菌PBS洗涤3次。随后,每孔加入10 µL CCK-8,继续孵育1 h。最后,用酶标仪测定450 nm波长处每孔的吸光值。
细胞毒性评价:利用公式,细胞存活率(%)=[实验孔吸光值-空白孔吸光值]/[对照孔吸光值-空白孔吸光值]×100%,进行细胞毒性评价。
本实施例细胞毒性评价结果见图3。如图3所示,在0-12 μM浓度范围内,绿原酸-小檗碱无载体纳米药物对小鼠乳腺上皮细胞抑制率小于5%,具有高度的生物相容性,不会对生物体产生毒副作用。
实施例4 绿原酸-小檗碱无载体纳米药物的抑菌活性分析
在本实施例中对实施例2制备得到的绿原酸-小檗碱无载体纳米药物的抑菌活性进行测定,方法如下:
采用96孔板微量稀释法,用于评价药物的抗菌活性。首先,每孔加入100 μL OD600=0.1的金黄色葡萄球菌或多重耐药金黄色葡萄球菌菌液。随后,依次加入100 μL苯唑西林钠、氨苄西林钠、绿原酸、小檗碱及绿原酸-小檗碱无载体纳米药物样品液,使各药物的终浓度分别为0.5、0.8、1.0、1.5、2.0、2.5 µM,充分混匀后在恒温培养箱(37℃)中培养16 h。最后,采用酶标仪测定600 nm波长处每孔的吸光值,以含细菌的营养肉汤为空白细菌对照,营养肉汤为溶剂对照。
参照以下公式计算各处理的细菌抑制率,细菌抑制率(%)=[实验孔吸光值-营养肉汤溶剂孔吸光值]/[空白细菌孔吸光值-营养肉汤溶剂孔吸光值]×100%。
本实施例中各处理的抑菌活性结果见图4。图4结果表明,实施例2制备的绿原酸-小檗碱无载体纳米药物具备优良的抗金黄色葡萄球菌活性,且对多重耐药金黄色葡萄球菌的抑菌活性明显优于目前诸多一线抗菌药物,例如苯唑西林、氨苄西林,具有深入研究,进一步临床开发的价值。本实施例经试验分析得到,绿原酸-小檗碱无载体纳米药物对多重耐药金黄色葡萄球菌的最小抑菌浓度(MIC)为1.5 µM。
实施例5 抗青霉素类耐药菌的药物组合物对多重耐药金黄色葡萄球菌的抑菌活性分析
在本实施例中对实施例2制备得到的抗青霉素类耐药菌的药物组合物进行抑菌活性测定,方法如下:
以多重耐药金黄色葡萄球菌为受试菌,选择1/2最小抑菌浓度(minimuminhibitory concentration,MIC)即0.75 µM绿原酸-小檗碱无载体纳米药物,与50 µM氨苄青霉素联用,观察其对多重耐药金黄色葡萄球菌生长的影响。试验分为空白对照组(仅含菌液不加药液)、组合物组(0.75 µM绿原酸-小檗碱无载体纳米药物+50 µM氨苄青霉素)、绿原酸-小檗碱无载体纳米药物组(0.75 µM)、氨苄青霉素组(50 µM)。以不加菌液仅加同浓度药液的为阴性对照组,用于消除药物颜色干扰。在37℃、180 r/min条件下振荡培养24 h,每4h无菌取样一次,采用酶标仪测定600 nm波长处的吸光值(OD600),其中试验处理组的OD600为试验处理组与同浓度同时段阴性组的吸光度之差,具体见表2。以时间为横坐标,OD600为纵坐标,绘制细菌生长曲线,见图5,分析药物对细菌生长的影响。
表2 抗青霉素类耐药菌的药物组合物的抑菌率表
图5和表2的结果表明,绿原酸-小檗碱无载体纳米药物组(0.75 µM)与空白对照组(即无药物处理组)在2 h后均进入细菌生长对数期,在4-12 h后绿原酸-小檗碱无载体纳米药物组(0.75 µM)未见明显抑菌效果。50 µM氨苄青霉素药物处理组相对于对照组,在2-8 h间表现出显著的抑菌效果,细菌呈现出迟缓增殖的趋势,但在10-12 h后的OD值(即细菌数量)明显趋近于对照组,抑菌率仅有11.08%,说明50 µM氨苄青霉素对多重耐药金黄色葡萄球菌表现出轻微的抑制作用,但仍无法有效杀菌。当组合物组(0.75 µM绿原酸-小檗碱无载体纳米药物+50 µM氨苄青霉素)处理时,多重耐药金黄色葡萄球菌在2-12 h内未出现迅速增殖现象,细菌抑制率>50%,呈现出较强杀菌效果,12 h后的OD值显著小于对照组,仅有极少量的多重耐药金黄色葡萄球菌生长,说明本实施例的组合物即0.75 µM绿原酸-小檗碱无载体纳米药物+50 µM氨苄青霉素对多重耐药金黄色葡萄球菌具有较好的杀菌作用,说明本发明制备的绿原酸-小檗碱无载体纳米药物能够协同氨苄青霉素,对多重耐药金黄色葡萄球菌发挥较强的抗菌作用。
实施例6 绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的耐药基因调控测定
在本实施例中对实施例2制备得到的绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的耐药基因调控作用进行测定,方法如下:
将对数生长期的多重耐药金黄色葡萄球菌(MRSA)稀释成终浓度为OD600=0.1的菌悬液,分为两组进行试验。以不加药物的菌悬液为空白对照组,0.75 µM 绿原酸-小檗碱无载体纳米药物处理的菌悬液为试验组,空白对照组和试验组的菌悬液于37℃恒温振荡培养8 h后。菌悬液在4℃条件下以8000 rpm 的转速离心10 min,收集菌泥用细菌RNA提取试剂盒进行提取后,利用RT-PCR试剂盒进行核酸浓度测定。
结果如图6所示,本发明的实施例2制备的绿原酸-小檗碱无载体纳米药物可显著抑制多重耐药金黄色葡萄球菌中抗青霉素类耐药相关基因erm、mecA及pbpB的表达(P<0.05),进而使青霉素类抗生素如氨苄青霉素重新表现出强大的抗菌活性,使本发明的实施例2制备的抗青霉素类耐药菌的药物组合物具有较好的杀菌效果。
实施例7 绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的生物被膜相关基因的影响
利用实施例2制备得到的绿原酸-小檗碱无载体纳米药物在亚抑菌浓度(0.75µM)下对金黄色葡萄球菌(S. aureus)和青霉素类耐药菌(MRSA)进行处理,并测定处理前后金黄色葡萄球菌和青霉素类耐药菌的与生物被膜相关基因(例如:agrA、sarA、icaA、cidA)表达量的变化,具体方法参照实施例6,测定结果见图7。
结果表明,与空白对照相比,绿原酸-小檗碱无载体纳米药物处理组的S. aureus和MRSA生物被膜相关基因(agrA、sarA、icaA、cidA)表达量均显著降低,说明本发明制备的绿原酸-小檗碱无载体纳米药物通过下调S. aureus和MRSA的agrA、sarA、icaA、cidA基因的表达量,从而降低MRSA的耐药性,进而使组合药物中的氨苄青霉素恢复其强大的抗菌活性,使本发明的实施例2制备的抗青霉素类耐药菌的药物组合物具有较好的杀菌效果。
实施例8 绿原酸-小檗碱无载体纳米药物对青霉素类耐药菌的营养吸收及核苷酸代谢相关基因表达的影响
金黄色葡萄球菌和青霉素类耐药菌的hrtA和hrtB 基因被认为是细菌吸收营养物质的关键转运系统,是细菌质膜的组成部分;deoD基因参与核苷酸代谢过程;蛋白质转位酶亚基SecE可以影响蛋白质的生物合成;rpmG3 是一个编码50S核糖体蛋白L33的基因,在核糖体的结构成分中发挥了重要作用;hla基因用于编码α-溶血素,α-溶血素是一种能使宿主红细胞和血小板细胞等细胞膜形成孔道的毒力蛋白,导致细胞溶解并造成溶血。
利用实施例2制备得到的绿原酸-小檗碱无载体纳米药物在亚抑菌浓度(0.75µM)下对金黄色葡萄球菌(S. aureus)和青霉素类耐药菌(MRSA)进行处理,并测定处理前后金黄色葡萄球菌和青霉素类耐药菌的deoD、hrtA、hrtB、SecE、rpmG3及hla基因表达量的变化,具体方法参照实施例6,测定结果见图8。
如图8所示,绿原酸-小檗碱无载体纳米药物能显著下调deoD、hrtA、hrtB、SecE、 rpmG3及hla基因在S. aureus和MRSA的表达水平,从而通过抑制S. aureus和MRSA的核苷酸的代谢、蛋白质和核糖体的生物合成以及破坏细菌细胞的表面结构等发挥其抗菌性能,提高制备的药物组合物对多重耐药金黄色葡萄球菌的杀菌能力。
青霉素类抗生素如氨苄青霉素,是通过与细菌的青霉素结合蛋白(penicillinbinding protein,PBP)相结合,进而阻止细菌的肽聚糖的合成和细菌的生长并最终发挥杀菌作用。青霉素类耐药菌对氨苄青霉素的耐药性,主要是通过两个方面进行的:第一是MRSA在菌体外形成生物被膜,第二是MRSA通过表达mecA基因,降低PBP与青霉素类抗生素的结合亲和力,使得青霉素类抗生物无法与PBP蛋白结合,进而失去抗菌活性。
综上所述,本申请制备的绿原酸-小檗碱无载体纳米药物能够显著降低金黄色葡萄球菌和多重耐药金黄色葡萄球菌的生物被膜以及耐药相关基因的表达水平,进而降低了金黄色葡萄球菌和多重耐药金黄色葡萄球菌对氨苄青霉素的耐药性,使氨苄青霉素恢复对金黄色葡萄球菌及多重耐药金黄色葡萄球菌的杀菌活性。本发明制备的抗青霉素类耐药菌的药物组合物,在绿原酸-小檗碱无载体纳米药物与氨苄青霉素的协同增效作用下,对青霉素类耐药菌表现出强大的抗菌活性。
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 (2)
1.制备抗青霉素类耐药菌的绿原酸-小檗碱纳米药物的方法,其特征在于,所述方法包括以下步骤:
(1)称取绿原酸与小檗碱分别溶解于甲醇中,使用氢氧化钠溶液调整绿原酸的甲醇溶液的pH值为7.0-7.5,将上述两种溶液搅拌混匀,缓慢加入至磷酸缓冲液中恒温搅拌,便于绿原酸与小檗碱进行自组装,得到混合溶液,所述小檗碱与绿原酸的摩尔比为1:1,所述氢氧化钠溶液的浓度为5 mg/mL,所述磷酸缓冲溶液为60℃的磷酸缓冲溶液;
(2)将步骤(1)的混合溶液置于透析袋内,采用超纯水透析24h以除去非自组装的小檗碱、绿原酸以及有机溶剂,每4h换一次超纯水,得到绿原酸与小檗碱自组装溶液,所述透析袋的截留分子量为3.5 kDa;
(3)将步骤(2)制得的绿原酸与小檗碱自组装溶液离心,离心转速为12000 rpm,离心时间为10-20 min,收集沉淀,超纯水洗涤3次,真空冷冻干燥,得到绿原酸-小檗碱纳米药物。
2.抗青霉素类耐药菌的耐药性的药物组合物,其特征在于,所述药物组合物为氨苄青霉素和权利要求1所制备的抗青霉素类耐药菌的绿原酸-小檗碱纳米药物,所述氨苄青霉素与所述绿原酸-小檗碱纳米药物复配的摩尔比为:200:3,所述青霉素类耐药菌为携带青霉素类耐药基因erm、mecA及/或pbpB基因的多重耐药金黄色葡萄球菌。
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