CN116270525A - 一种用于治疗肺部细菌感染性肺炎的可吸入仿生纳米材料及其制备方法和应用 - Google Patents
一种用于治疗肺部细菌感染性肺炎的可吸入仿生纳米材料及其制备方法和应用 Download PDFInfo
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Abstract
本发明提供了一种用于治疗肺部细菌感染性肺炎的可吸入仿生纳米材料及其制备方法和应用,属于药物制备技术领域。以嵌段式聚醚和3,3',5,5'‑四甲基联苯胺为模板通过一锅合成法制备介孔聚多巴胺纳米颗粒A;将左氧氟沙星加入A中,搅拌离心,冷冻干燥,得到纳米颗粒B;将RAW264.7细胞,离心后重悬,匀浆后再次离心收集上清,上清离心沉淀即为制备的巨噬细胞膜C;将配置的B溶液和C混合,采用直接挤压法制备包裹巨噬细胞膜涂层的仿生纳米材料D。本发明制得可吸入仿生纳米材料,能够跨越复杂的肺屏障靶向炎症部位以应答释放药物,并能清除活性氧缓解肺部炎症,对肺部感染治疗意义重大且具有推广应用价值。
Description
技术领域
本发明涉及药物制备技术领域,具体涉及一种用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料及其制备方法和应用。
背景技术
下呼吸道感染(LRTIs)是近年来导致全球人类死亡的主要原因之一。其中,急性下呼吸道感染已成为儿童住院和死亡的主要原因,占全球儿童死亡人数的15%。肺炎链球菌、铜绿假单胞菌和金黄色葡萄球菌等细菌,是肺炎中重要的病原体。对于细菌性肺炎的治疗,吸入给药可以直接将抗生素输送到感染区域,具有高的依从性和安全性,因此近年来受到了越来越多的关注。但是由于下呼吸道特殊的生理和病理结构,仍存在许多问题阻碍吸入给药的应用。如黏液层作为抗生素的物理屏障,降低了吸入治疗的疗效;肺泡巨噬细胞易吞噬外来物质,限制了肺炎的治疗效率。微纳米颗粒(NPs)已被研究用于装载抗生素,通过吸入方式将其送入肺部,但其克服复杂的肺部屏障的能力仍有待优化。因此,具有仿生靶向性的药物传递系统对吸入式给药治疗肺部感染具有重要意义。
此外,持续的炎症反应产生过量的活性氧,其可以直接破坏核酸、蛋白质和脂质,对机体产生不可逆的破坏,如果错过最佳治疗时间,活性氧和炎症反应将启动一个相互促进的循环,并全身传播,从而导致全身炎症综合征。随着全身炎症综合征的发生,过量的活性氧会选择性地攻击主要器官,导致器官功能障碍,甚至死亡。因此,如何在保证有效清除肺炎链球菌的同时改善肺部炎症环境是临床待解决的重要问题。
目前,未见通过可吸入仿生纳米材料来克服复杂的肺屏障应答释放药物并有效清除活性氧缓解肺部炎症的制备方法的报道。
发明内容
针对现有技术中存在的上述问题,本发明目的之一提供一种可克服复杂的肺屏障并有效缓解肺部炎症的的仿生药物传递系统,本发明目的之二提供一种可克服复杂的肺屏障并有效缓解肺部炎症的的仿生药物传递系统的制备方法,本发明目的之三通过仿生靶向递送,以避免肺屏障对药物的清除,实现有效抗菌并同时清除ROS治愈细菌引起的局部炎症。
本发明提供以下技术方案:
一种用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,包括以下步骤:
将嵌段式聚醚和3,3',5,5'-四甲基联苯胺溶解在水和乙醇的混合物中,搅拌后加入120mg多巴胺盐酸盐,搅拌反应后离心并收集沉淀,产物采用溶剂萃取法去除模板,得到纳米颗粒A;将纳米颗粒A与左氧氟沙星溶解于PBS中,搅拌反应后离心收集沉淀物,制备含有左氧氟沙星的纳米颗粒B;取RAW264.7细胞,离心后的沉淀重悬匀浆后再次离心收集上清,上清离心沉淀后得到巨噬细胞膜C;将巨噬细胞膜C与纳米颗粒B混合,在冰浴中超声处理;采用直接挤压法制备得到含巨噬细胞膜涂层的仿生纳米材料。
进一步的,所述嵌段式聚醚和3,3',5,5'-四甲基联苯胺的质量比为(1:1)~(1:3);所述纳米颗粒A与左氧氟沙星的质量比为(2:1)~(1:2)。
本发明提供一种用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,包括以下步骤:
1)将嵌段式聚醚(F127,0.6-0.8g)和3,3',5,5'-四甲基联苯胺(TMB,1.0-1.2g)(质量比1:1~1:3)溶解在水和乙醇的混合物中,搅拌30min;然后,加入120mg多巴胺盐酸盐,在室温下搅拌反应一定时间,离心,收集沉淀,产物用乙醇和丙酮洗涤数次,并采用溶剂萃取法去除模板,制备纳米颗粒A;
2)将纳米颗粒A(30-50mg)与左氧氟沙星(30-50mg)(质量比2:1~1:2)溶解于PBS(pH 7.4)中,搅拌12h;随后,离心收集沉淀物,制备含有左氧氟沙星的纳米颗粒B;
3)取RAW264.7细胞(该细胞系由中国科学院细胞库提供),800g离心5min,沉淀用含1mM苯甲基磺酰氟(PMSF)的低渗缓冲液(NaHCO3和乙二胺四乙酸组成)重悬,用匀浆去核器上下20次,在4℃,3200g下离心收集上清,上清在15000g离心沉淀即为制备的巨噬细胞膜C;
4)将巨噬细胞膜C与纳米颗粒B按质量比1:1混合,在冰浴中超声处理一定时间;然后采用直接挤压法制备含巨噬细胞膜涂层的仿生纳米材料D。
优选的,所述F127和TMB的质量为1:1.5。
优选的,所述水和乙醇的体积比为13:12。
优选的,巨噬细胞膜C与纳米颗粒B在冰浴中超声处理的时间为1~3min。
优选的,所述巨噬细胞膜C与纳米颗粒B在冰浴中超声处理的时间为2min。
优选的,所述纳米颗粒A与左氧氟沙星质量比为1:1。
优选地,所述巨噬细胞膜C与纳米颗粒B混合后在冰浴中超声处理功率为80~120W,超声1~3s,停止4~6s。
优选的,所述巨噬细胞膜C与纳米颗粒B混合后在冰浴中超声处理功率为100W,超声2s,停止5s。
本发明还提供上述制备方法制得的可吸入仿生纳米材料。
优选的,所述可吸入仿生纳米材料的水合粒径为100~150nm。
本发明还提供上述制备方法制得的可吸入仿生纳米材料在药物递送方面的应用。
本发明还提供上述可吸入仿生纳米材料在制备用于肺部菌感染性肺炎治疗药物中的应用。
优选的,所述负载的药物包括普鲁卡因青霉素、哌拉西林、苄星青霉素、阿莫西林、美洛西林、阿莫西林克拉维酸钾、左氧氟沙星或头孢菌素类药物。
优选的,肺部菌感染性肺炎的菌株包括肺炎链球菌、金黄色葡萄球菌或铜绿假单胞菌。
本发明构建一个具有克服复杂肺屏障能力高效清除肺炎病原菌和抑炎性能的仿生可吸入纳米材料(LE@MPDA-MM)。本发明以肺炎链球菌感染的模型为例,肺炎链球菌是全球肺炎死亡的主要原因。2016年,它造成的死亡人数超过了所有其他肺炎原因造成的死亡人数之和,尽管人们广泛使用抗菌剂及抗生素用来治疗肺炎链球菌感染,但是肺部肺泡巨噬细胞吞噬严重限制了肺炎链球菌感染性肺炎的治疗效率。考虑到巨噬细胞的炎症趋向性和跨越屏障的能力,本发明通过仿生策略利用巨噬细胞膜表面的糖基基团及天然的炎症趋向性来驱动载体有效靶向并积累在肺部炎症部位以高效的清除肺炎链球菌;此外由肺部炎症引起的过量ROS通过直接破坏核酸、蛋白质和脂质,对机体产生不可逆的破坏,也是待解决的问题。因此,本发明设计了具有pH响应性及清除ROS能力的仿生纳米递送载体,既能有效的响应肺部感染部位微酸环境释放药物,又能清除活性氧,减少肺部感染,缓解炎症反应。与目前的肺部感染治疗手段不同,本发明设计简单,材料安全,既能跨越屏障有效递送抗生素杀灭细菌避免耐药性等问题,同时又能减少肺部感染,缓解炎症反应,对肺部感染治疗具有重要意义。
本发明制得的可吸入仿生纳米材料可用于肺部炎症部位药物的有效递送及响应释放,所述的肺炎感染菌株包括肺炎链球菌,金黄色葡萄球菌,铜绿假单胞菌。
有益效果
1)本发明的可吸入仿生纳米材料,合成材料来源丰富,方便获取,成本低,合成简单,时间周期短,便于临床转化。
2)本发明提供的可吸入仿生纳米材料,借助巨噬细胞固有的性质与炎症定向趋化能力避免被肺部肺泡巨噬细胞吞噬,有效地驱动载体在肺部炎症组织中的积累。
3)本发明提供的可吸入仿生纳米材料,包含的介孔聚多巴胺一方面具有高的载药率,可以有效地包封抗生素药物;另一方面该纳米颗粒含有较多的氨基基团,具有pH敏感性,在酸性条件下促进药物的释放;此外,纳米颗粒富含邻苯二酚官能团,能有效的消除活性氧,抑制炎症,减轻肺组织病理学损伤,具有推广应用价值。
附图说明
图1实施例1制得的可吸入仿生纳米材料的电位与粒径分析图(图A为电位,图B为粒径);
图2实施例1制得的可吸入仿生纳米材料的透射电镜图;
图3实施例1制得的可吸入仿生纳米材料载药效率图;
图4实施例1制得的可吸入仿生纳米材料的pH响应性释放动力学图;
图5实施例1制得的可吸入仿生纳米材料对ROS清除能力图;
图6实施例1制得的可吸入仿生纳米材料处理LPS刺激的Raw264.7细胞后炎症因子水平图(图A为TNF-α,图B为IL-6);
图7实施例1制得的可吸入仿生纳米材料对菌活力影响图(图A为菌液的OD600值与LE@MPDA-MM的浓度关系;图B为菌液的OD600值与时间的关系);
图8实施例1制得的可吸入仿生纳米材料体外抗吞噬能力图;
图9实施例1制得的可吸入仿生纳米材料跨越粘液屏障能力图;
图10实施例1制得的可吸入仿生纳米材料体内杀菌效果图(图A为平板照片,图B为细菌菌落总数计数结果);
图11实施例1制得的可吸入仿生纳米材料体内抗炎效果图(图A为IL-6,图B为TNF-α);
具体实施方式
为使本发明的技术方案、优点和目的更加清楚,下面将结合本发明实施例的附图,对本发明实施例的技术方案进行清楚、完整地描述。显然,所描述的实施例是本发明的一部分实施例,而不是全部的实施例。基于所描述的本发明的实施例,本领域普通技术人员在无需创造性劳动的前提下所获得的所有其他实施例,都属于本申请的保护范围。
下面结合附图和实施例对本发明作进一步说明。
实施例1
一种可吸入仿生纳米材料的制备方法,包括以下步骤:
1)将0.72g F127和1.08g TMB溶解在水(130mL)和乙醇(120mL)的混合物中,搅拌30min;然后,加入120mg多巴胺盐酸盐,在室温下搅拌24h,离心,产物用乙醇和丙酮洗涤,并在乙醇和丙酮混合溶液(2:1v/v)中超声处理三次(每次30min)去除模板,制备纳米颗粒A;
2)将纳米颗粒A溶液(40mg,8mL)与盐酸左氧氟沙星溶液(LE,40mg,8mL)混合,搅拌12h;随后,离心收集沉淀物,制备含有左氧氟沙星的纳米颗粒B;
3)取RAW264.7细胞,800g离心5min,沉淀用含1mM苯甲基磺酰氟(PMSF)的低渗缓冲液(NaHCO3和乙二胺四乙酸组成)重悬,用匀浆去核器上下20次,在4℃,3200g下离心收集上清,上清在15000g离心沉淀即为制备的巨噬细胞膜C;
4)将巨噬细胞膜C与纳米颗粒B混合,在冰浴中超声处理2min(100W,超声2s,停止5s);然后采用直接挤压法制备包裹巨噬细胞膜涂层的仿生纳米材料D;
实施例2
将实施例1中制得的仿生喷雾剂用于治疗肺炎链球菌感染性肺炎。
具体为:将肺炎链球菌感的菌落转移到10mL的THB培养基中,在37℃中培养过夜,获得处于指数生长期的细菌。
检测与分析
1)将实施例1制得的可吸入仿生纳米材料进行流体动力学分析电位水平及粒径尺寸分布情况,结果如图1所示。
从图1中分析可知,含有左氧氟沙星的聚多巴胺纳米材料(LE@MPDA)的平均尺寸约为90nm,巨噬细胞膜(MMs)涂层后,平均尺寸增加至约138nm,LE@MPDA的电位为-32.2mV,MM的电位为-23.4mV,在LE@MPDA表面涂覆MMs后,LE@MPDA-MM电位与巨噬细胞膜相似,为-22.2mV,这表明MMs成功覆盖在LE@MPDA的表面。
2)将实施例1制得的可吸入仿生纳米材料进行透射电子显微镜分析,比例尺为50nm,结果如图2所示。
从图2中观察分析可知,仿生纳米材料(LE@MPDA-MM)具有球形结构,平均直径约为127nm。
3)将实施例1制得的可吸入仿生纳米材料进行药物负载效率的考察
具体操作为:取实施例1制得的可吸入仿生纳米材料1mg,加入无水甲醇溶液5mL,高速涡旋10min进行颗粒破坏,随后离心并收集上清。通过LE标准曲线,计算上清中LE量,并测定载药率(LE),测定如下:
LE(%)=WA/(WA +WB)×100
WA为LE的质量,WB为总载体的质量,WC为总LE的质量。
由于聚多巴胺具有多空的结构特点,载药率高达12%左右,这为高效清除肺炎部位病原菌奠定了基础(图3)。
4)将实施例1制得的可吸入仿生纳米材料进行响应pH释药分析
具体操作为:为探讨可吸入仿生纳米材料的药物释放行为,取1mg可吸入仿生纳米材料(LE@MPDA-MM)分散在5mL不同溶剂中,将含有1mL LE@MPDA-MM溶液的透析袋(MWCO:8-14kDa)浸入10mL磷酸盐缓冲液(0.1M,pH 7.4,含0.5%tween-80;或者0.1M,pH 6.0,含0.5%tween-80)中,在不同的时间间隔内取2mL溶液,并用等量新鲜相同pH值的PBS代替,在293nm处用HPLC定量LE的累积释放量。
结果显示,LE@MPDA-MM在pH 7.4PBS介质中24h累计释放效率仅为22%左右,而在pH 6.0PBS中的24h累积释放73%左右,是pH 7.4PBS释放介质得3倍多,这是因为在低pH条件下,MPDA分子中氨基的质子化作用增强,MPDA和LE之间的π-π相互作用减弱,从而增加了药物的释放。以上结果表明,LE@MPDA-MM可在炎症弱酸性环境下能够快速释放药物,从而有助于在炎症部位消除细菌(图4)。
5)将实施例1制得的可吸入仿生纳米材料进行ROS清除能力的考察
具体操作为:将Mlg细胞(2×105)接种于6孔板中过夜,将含有200μM过氧化氢的无血清培养基替代旧培养基,孵育1.5h;然后,在六孔板中分别加入MPDA NPs、LE@MPDA NPs和LE@MPDA-MM NPs,继续孵育4h,以未经制剂处理组为阳性对照,以无过氧化氢处理的细胞为阴性对照;随后,加入DCFH-DA检测自由基水平,并用Hoechst染色;最后,用荧光显微镜观察细胞。
如图5所示,与阳性对照相比,经含有MPDA处理后的Mlg细胞的荧光强度明显降低,证实了MPDA清除自由基的能力。
6)将实施例1制得的可吸入仿生纳米材料进行抗炎性能的考察
具体操作为:为了检测仿生纳米材料的抗炎能力,将RAW 264.7细胞接种于6孔板中过夜,采用LPS(1μg/mL,24h)诱导炎症环境,实验组1:空白培养基(不含LPS),设置为阴性对照;实验2组:LPS,设置为阳性对照组;实验3组:LPS+MPDA NPs;实验4组:LPS+LE@MPDANPs;实验5组:LPS+LE@MPDA-MM NPs,按照分组分别处理细胞,并孵育4h;然后,用Elisa试剂盒检测上清液中的IL-6和TNF-α,结果如图6所示。
结果显示,含MPDA的制剂处理后,已被LPS刺激的RAW 264.7细胞IL-6和TNF-α分泌水平显著降低,说明含MPDA的仿生纳米材料可以有效地抑制炎症。
7)将实施例1制得的可吸入仿生纳米材料进行抗菌能力的考察
最低抑制浓度的考察:为了考察最低抑菌浓度MIC,将肺炎链球菌落转移到10mL的THB培养基中,在37℃的培养箱培养过夜,获得处于指数生长期的细菌,稀释菌悬液浓度至2×105CFU/mL,于96孔板中,将一定量的LE@MPDA-MM悬液与等细菌悬液进行混合,样品中LE的最终浓度范围为1~16μg/mL。所有96孔板在37℃下温和培养12h,通过酶标仪OD600(Bio-RoadModel680,UK)测定MIC。
考察仿生纳米材料的体外抗菌效果:将细菌悬浮按照1:100的比例加到THB培养基里面,以LE@MPDA-MM的MIC为考察浓度,37℃下温和培养,检测不同时间点OD600的变化情况,以评估仿生纳米材料的体外抗菌效果。
结果显示,当LE@MPDA-MM的浓度仅为8.5μg/mL时,就表现出明显的抑菌效果。在浓度为34μg/ml时,菌液的OD600值和空白组(纯培养基)的OD600值几乎接近,因此LE@MPDA-MM组肺炎链球菌的MIC值为34μg/mL(图7A)。此外,体外抗菌实验也再次表明了LE@MPDA-MM具有优异的抗菌效果,培养数十个小时后,OD600值始终接近空白组。
8)仿生纳米材料体外抗吞噬性能的评价
本研究以RAW264.7为模型来计算仿生纳米系统的体外抗吞噬性能,将RAW 264.7细胞以每孔105个细胞密度接种于12孔板中,培养24h;事先用脂质体包覆LE@MPDA形成LE@MPDA-lip,然后用DIO染料对LE@MPDA-lip和LE@MPDA-MM进行染色,最后与上述细胞共孵育4h,通过流式细胞术评价抗吞噬能力。
如图8显示,LE@MPDA-MM处理的巨噬细胞的荧光水平低于LE@MPDA组,这主要由于MM涂层可以有效地阻断巨噬细胞的非特异性吞噬。
9)考察仿生纳米材料跨粘液屏障的性能
人工黏液的制备:将500mg脱氧核糖核酸,250mg粘蛋白,0.295mg二乙烯三胺五乙酸,1mL RPMI 1640氨基酸溶液,250μL蛋黄乳液,250mg氯化钠和110mg氯化钾混合并溶解于50mL的无DNase水中,在25℃下平衡2h备用。
仿生纳米材料跨粘液屏障的性能的考察:实验分组,实验1组:LE@MPDA组;实验2组:LE@MPDA-MM。将人工黏液在硬化的明胶凝胶上;然后,将样品用考马斯亮蓝染色,并沉积在人工黏液层的表面。分别记录样品在0min、10min、30min、60min和120min时的图像。
图9结果显示,相比于LE@MPDA,LE@MPDA-MM在黏液中的具有高的扩散速度,表明MM涂层在克服黏液阻碍方面发挥了重要的作用。
10)考察仿生纳米材料体内抗菌及抑炎性能
鼠肺感染模型的构建:选择雌性BALB/c小鼠(8周龄,18-20g),在异氟醚麻醉下,采用滴鼻液法鼻内接种肺炎链球菌,2×108CFU/小鼠,接种后的小鼠保持直立5min以建立小鼠肺炎感染模型,实验1组:鼻内接种生理盐水的小鼠作为control组,即“健康对照”组;实验2组:PBS组,作为阳性对照;实验3组:LE;实验4组:LE@MPDA;实验5组:LE@MPDA-MM。
体内抗菌性能的考察:在细菌接种治疗后,处死小鼠,用400μL生理盐水冲洗肺2次,收集小鼠肺,将肺组织均质化。将匀浆右肺稀释104倍,涂布于TSA培养基上。样品在37℃下培养过夜,最后观察平板上细菌生长情况。
体内抑炎性能的考察:处理小鼠后,检测小鼠支气管肺泡灌洗液(BAL)中炎症因子的水平(TNF-α、IL-6和IL-1β)。
图10结果显示,在肺匀浆中平板中,相比于LE组,LE@MPDA-MM组基本上观察不到明显的菌落,与细菌菌落总数计数结果相同,这说明LE@MPDA-MM吸入给药可有效克服黏液渗透能力和应答药物释放特性,高效的消除肺炎链球菌。
图11结果显示,经吸入治疗后,LE@MPDA组和LE@MPDA-MM组的肺灌洗液中的TNF-α和IL-6显著低于其他组,说明含MPDA组的细菌性肺炎的炎症症状得到改善,表明含有MPDA制剂组经吸入治疗后,可有效减少肺部感染,缓解炎症反应。
本发明构建了一个具有克服黏液屏障能力和抑炎性能的仿生可吸入纳米材料(LE@MPDA-MM)。本发明以肺炎链球菌感染的模型为例,肺炎链球菌是全球肺炎死亡的主要原因。2016年,它造成的死亡人数超过了所有其他肺炎造成的死亡人数之和,尽管人们广泛使用抗菌剂及抗生素用来治疗肺炎链球菌感染,但是呼吸道的粘液及肺部肺泡巨噬细胞吞噬严重限制了肺炎链球菌感染性肺炎的治疗效率。考虑到巨噬细胞的炎症趋向性和跨越屏障的能力,本发明通过仿生策略利用巨噬细胞膜表面的糖基基团及天然的炎症趋向性来驱动载体有效靶向并积累在肺部炎症部位以高效的清除肺炎链球菌;由肺部炎症引起的过量ROS通过直接破坏核酸、蛋白质和脂质,对机体产生不可逆的破坏,因此,本发明设计具有pH响应性及清除ROS能力的聚多巴胺纳米颗粒装载抗生素,既能有效的响应肺部感染部位微酸环境释放药物,又能清除活性氧,减少肺部感染,缓解炎症反应。与目前的肺部感染治疗手段不同,本发明设计简单,材料安全,既能跨越屏障有效递送抗生素杀灭细菌,同时又能减少肺部感染,缓解炎症反应,对肺部感染治疗具有重要意义。
在实际制备和研究过程中,还发现,通过改变MPDA的合成投料比可以有效调节纳米材料的大小,为有效的肺部药物递送提供了强有力的技术支撑。
最后说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本技术方案的宗旨和范围,其均应涵盖在本发明的保护范围当中。
Claims (10)
1.一种用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,包括以下步骤:
将嵌段式聚醚和3,3',5,5'-四甲基联苯胺溶解在水和乙醇的混合物中,搅拌后加入多巴胺盐酸盐,搅拌反应后离心并收集沉淀,产物采用溶剂萃取法去除模板,得到纳米颗粒A;将纳米颗粒A与左氧氟沙星溶解于PBS中,搅拌反应后离心收集沉淀物,制备含有左氧氟沙星的纳米颗粒B;取RAW264.7细胞,离心后的沉淀重悬匀浆后再次离心收集上清,上清离心沉淀后得到巨噬细胞膜C;将巨噬细胞膜C与纳米颗粒B混合,在冰浴中超声处理;采用直接挤压法制备得到含巨噬细胞膜涂层的仿生纳米材料。
2.根据权利要求1所述用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,所述嵌段式聚醚和3,3',5,5'-四甲基联苯胺的质量比为(1:1)~(1:3);所述纳米颗粒A与左氧氟沙星的质量比为(2:1)~(1:2)。
3.根据权利要求1所述用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,水和乙醇的体积比为13:12。
4.根据权利要求1所述用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,巨噬细胞膜C与纳米颗粒B在冰浴中超声处理的时间为1~3min。
5.根据权利要求1所述用于治疗肺部菌感染性肺炎的可吸入仿生纳米材料的制备方法,其特征在于,所述巨噬细胞膜C与纳米颗粒B混合后在冰浴中超声处理功率为80~120W,超声1~3s,停止4~6s。
6.一种可吸入仿生纳米材料,其特征在于,由权利要求1~5任一所述方法制备而成。
7.根据权利要求6所述的可吸入仿生纳米材料,其特征在于,所述可吸入仿生纳米材料的粒径为100~150nm。
8.权利要求7所述的可吸入仿生纳米材料在制备用于肺部菌感染性肺炎治疗药物中的应用。
9.根据权利要求8所述可吸入仿生纳米材料在肺部菌感染性肺炎治疗中的应用,其特征在于,所述可吸入仿生纳米材料负载的药物包括普鲁卡因青霉素、哌拉西林、苄星青霉素、阿莫西林、美洛西林、阿莫西林克拉维酸钾、左氧氟沙星或头孢菌素类药物。
10.根据权利要求9所述可吸入仿生纳米材料在肺部菌感染性肺炎中的应用,其特征在于,肺部菌感染性肺炎的菌株包括肺炎链球菌、金黄色葡萄球菌或铜绿假单胞菌。
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