CN111269367A - 一种酸响应电荷屏蔽纳米粒子及其制备方法和应用 - Google Patents
一种酸响应电荷屏蔽纳米粒子及其制备方法和应用 Download PDFInfo
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- CN111269367A CN111269367A CN202010070244.2A CN202010070244A CN111269367A CN 111269367 A CN111269367 A CN 111269367A CN 202010070244 A CN202010070244 A CN 202010070244A CN 111269367 A CN111269367 A CN 111269367A
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Abstract
本发明属于药物化学和生物可降解医用高分子领域,公开了一种酸响应电荷屏蔽聚合物及其制备和应用。本发明利用二甲基马来酸酐修饰胍基,使其在正常环境下被屏蔽。且该聚合物自组装形成纳米粒子的同时可以装载不同的小分子抗生素,光热剂,光敏剂等药物形成载药纳米粒子,当载药纳米粒子达到细菌生物膜感染部位时,由于感染部位呈现酸性环境,保护基团二甲基马来酸酐在感染部位脱保护,暴露胍基基团,展现出纳米粒子穿透细菌生物膜的能力。载药纳米粒子结合光热治疗,光动力治疗或化学药物治疗,能大幅度提高纳米药物的治疗效果且不会使细菌产生耐药性。
Description
技术领域
本发明属于药物化学和生物可降解医用高分子领域,特别涉及一种酸响应电荷屏蔽聚合物及其制备和应用。
背景技术
胍基是一种易溶于水,呈现碱性的化学基团,其化学式为-CN3H4。现如今胍基基团已经广泛应用于深穿透抗肿瘤治疗领域,其作用机制可以简单描述为:首先,胍基基团带有大量的正电荷,可以与带有负电荷的细胞膜紧密结合,进而通过氢键的作用力穿透并进入细胞内部;大量的文献证明,胍基可以与蛋白质上的某些化学键产生相互作用,使得胍基具有结合蛋白质的功能。目前,随着多药耐药细菌的出现,以及人们对抗菌肽的研究逐渐深入,发现大部分抗菌肽中均含有大量的精氨酸残基,这些氨基酸残基含有大量的胍基基团,这一发现也进一步验证了胍基是具有很好抗菌功能的。
目前,由于多药耐药菌的出现,使得细菌感染所带来的疾病更加难以治疗,并且细菌在感染部位极易形成细菌生物膜,这使得传统药物更加难以渗透进入细菌深层感染部位,因此使得细菌生物膜感染难以治愈且容易复发。
在众多的研究中发现胍基基团由于其本身所具有的易穿透细胞膜的性质,使得胍基基团本身具有很强的生物毒性,裸露的胍基在血液循环中十分容易造成血细胞的破碎,引起溶血现象的发生,导致极强的副作用。这使得胍基本身的应用受到极大的限制。
所以如何在血液循环中屏蔽胍基,提高药物的生物安全性,且可以在感染部位迅速释放胍基,提高药物渗透进入细菌生物膜并杀死细菌,甚至不会使得细菌产生耐药性显得十分有意义。
发明内容
为了克服上述现有技术的缺点与不足,本发明的首要目的在于提供一种具有屏蔽胍基的可以运载药物深层渗透进入细菌生物膜的酸响应电荷屏蔽聚合物。
本发明另一目的在于提供上述酸响应电荷屏蔽聚合物的制备方法。
本发明再一目的在于提供上述酸响应电荷屏蔽聚合物在制备载药纳米粒子中的应用。
本发明的目的通过下述方案实现:
一种酸响应电荷屏蔽聚合物,其由以下方法制备得到:
(1)将亲水单体和小分子链转移剂在引发剂存在条件下进行聚合,得到大分子链转移剂;
(2)将大分子链转移剂和疏水单体在引发剂存在条件下进行聚合,得到聚合物1;
(3)在催化剂存在条件下,将聚合物1和1H-吡唑-1-甲脒盐酸盐反应,得到富含胍基的聚合物2;
(4)在催化剂存在条件下,将富含胍基的聚合物2与马来酸酐反应,得到胍基被屏蔽的聚合物,即所述的酸响应电荷屏蔽聚合物。
步骤(1)中所述的亲水单体为氨基封端的亲水单体、N,N-二甲基丙烯酸胺(DMA)、乙二醇(PEG)、甲基丙烯酸二甲氨基乙酯(DMAEMA)等中的至少一种;其中氨基封端的亲水单体为2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl)、N-(3-氨基丙基)甲基丙烯酰胺盐酸盐、赖氨酸等中的至少一种;
优选的,步骤(1)中所述的亲水单体为2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl)和N,N-二甲基丙烯酸胺(DMA)的混合物。
步骤(1)中所述的引发剂为偶氮二异丁腈(AIBN)、偶氮二异庚腈(ABVN)、偶氮二异丁二甲酯(AIBME)中的一种;步骤(1)中聚合反应所用的溶剂为二甲基亚砜、二氯甲烷、1,4-二氧六环、无水乙醇和二甲基甲酰胺中的至少一种;
步骤(1)中所述的2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl)、N,N-二甲基丙烯酸胺(DMA)、小分子链转移剂以及引发剂的摩尔比为2~1200:2~1200:1~100:0.1~3;优选为181:272:4.5:1。
步骤(1)中所述的聚合是指在真空密封条件下60~80℃反应12~75h,优选为在70℃反应15h;
步骤(2)中所述的疏水单体为甲基丙烯酸酯(BMA)、苯乙烯,乙烯基三甲基硅烷等中的至少一种,优选为甲基丙烯酸酯(BMA);步骤(2)中所述的引发剂为偶氮二异丁腈(AIBN)、偶氮二异庚腈(ABVN)、偶氮二异丁二甲酯(AIBME)中的一种。
步骤(2)中所述的大分子链转移剂、疏水单体和引发剂的摩尔比为1~100:2~1200:0.2~3;优选为1.2:72:0.24。
步骤(2)中所述的聚合是指在真空条件下65~85℃反应12~24h,优选为在真空条件下70℃反应15h;聚合反应的溶剂为二甲基亚砜,二氯甲烷、1,4二氧六环中的至少一种;
步骤(3)中所述的催化剂为N,N-二异丙基乙胺(DIPEA)、三乙胺(TEA)中的至少一种;
步骤(3)中所述的催化剂、聚合物1和1H-吡唑-1-甲脒盐酸盐的摩尔比为6~800:0.1~10:3~500;
步骤(3)中所述的反应是指在溶剂存在和真空的条件下20℃~75℃搅拌反应12~72h,优选为在55℃反应12h;反应的溶剂为二甲基亚砜;
步骤(4)中所述的催化剂为N,N-二异丙基乙胺(DIPEA);
步骤(4)中所述的催化剂、富含胍基的聚合物2和马来酸酐的摩尔比为6~800:0.1~1:3~500;
步骤(4)中所述的反应是指在溶剂存在条件下常温反应12~72h,其中溶剂为二甲基亚砜、二氯甲烷中的至少一种;步骤(4)反应结束后还包括一个将所得反应液在弱碱性(pH7.4~9)的去离子水中透析(透析分子量为1000~5000kDa)的步骤。
上述的酸响应电荷屏蔽聚合物作为载体在制备载药纳米粒子中的应用。
一种载药纳米粒子,由以下方法制备得到:将具有抗菌活性的物质,如光敏剂、光热剂或抗生素等与酸响应电荷屏蔽的聚合物共同溶解在有机溶剂中,然后加入到正在搅拌(搅拌速度为500~2000rpm)的水中,继续搅拌0.5~12h,再通过透析或者超滤法除去有机溶剂,最终得到高效载药纳米粒子;或者,将具有抗菌活性的物质,如光敏剂、光热剂或抗生素等与酸响应电荷屏蔽的聚合物共同溶解在有机溶剂中,在搅拌的条件下(搅拌速度为500~2000rpm)向其中缓慢注入水,注水速度为每小时0.09~9mL,注水完毕后再通过透析或者超滤法去除有机溶剂,最终得到高效载药纳米粒子;
所述的光敏剂为吲哚菁绿、曙红Y、盐酸氨酮戊酸等中的至少一种,所述的光热试剂为长沙红、纳米金棒、聚吡咯等中的至少一种,所述的抗生素为阿莫西林、头孢哌酮、红霉素、庆大霉素等中的至少一种;
所述的有机溶剂为1,4-二氧六环、二甲基亚砜、二甲基甲酰胺中的至少一种;
所述的具有抗菌活性的物质与酸响应电荷屏蔽聚合物的质量比为1:1~100;所述的有机溶剂与水的体积比为1:9;所述的酸响应电荷屏蔽聚合物与水的质量体积比为0.1~100mg/L。
所述的透析是利用分子量为1kDa以上的透析膜在去离子水中进行透析;所述的超滤是指用使用分子量10kDa以上的超滤管,不断离心去除溶剂,再补充水溶液,重复10次以上即可。
本发明的机理为:
本发明以含有氨基基团的2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl),亲水性的N,N-二甲基丙烯酰胺(DMA)和疏水性的甲基丙烯酸丁酯(BMA)为原料,通过可逆加成断裂链转移聚合方法(RAFT)聚合得到聚合物,并对聚合物中的氨基进行修饰得到富含胍基的聚合物。进一步利用二甲基马来酸酐修饰胍基,使其在正常环境下被屏蔽。当纳米药物达到细菌生物膜感染部位时,在细菌生物膜感染及炎症反应的作用下,由于感染部位呈现酸性环境,保护基团二甲基马来酸酐(DA)在感染部位脱保护,暴露胍基基团,展现出纳米粒子穿透细菌生物膜的能力。该嵌段聚合物自组装形成纳米粒子的同时可以装载不同的小分子抗生素,光热剂,光敏剂等药物,其本身具有药物运输和特异靶向深穿透细菌生物膜的功能。纳米药物结合光热治疗,光动力治疗或化学药物治疗,能大幅度提高纳米药物的治疗效果且不会使细菌产生耐药性。
本发明相对于现有技术,具有如下的优点及有益效果:
本发明制备得到的酸响应电荷屏蔽纳米粒子在血液循环过程时,其本身不带任何电荷,不会损伤血细胞,具有良好的血液循环稳定性。而当纳米粒子到达细菌感染部位时,由于感染部位呈现弱酸性环境,导致纳米粒子保护基团脱保护暴露出胍基基团,进而使得纳米粒子携带有较强的正电荷,可以很好的与细菌生物膜结合并深层渗透进入细菌生物膜内部。纳米药物本身可以很好的与细菌粘附,进而穿刺细菌细胞膜,到达一定的物理杀菌效果。然后纳米粒子所携带的药物可以发挥相应的疗效,更好消灭细菌生物膜内部的细菌且不产生耐药性。
本发明主要解决了传统药物难以渗透进入细菌生物膜内部和含有胍基的纳米粒子其血液循环稳定性差的问题。此外,在保证了纳米粒子的稳定性后,提高了纳米粒子对细菌生物膜的渗透能力和对细菌的杀伤能力,且不会使得细菌产生耐药性。
附图说明
图1为实施例5制备得到的载药纳米粒子紫外吸收可见光谱图;
图2为实施例5制备得到的载药纳米粒子的粒径分布图;
图3为实施例4制备得到的酸响应电荷屏蔽聚合物和荧光胺的混合液在pH7.4的环境下荧光胺的荧光强度随时间的变化图;
图4为实施例4制备得到的酸响应电荷屏蔽聚合物和荧光胺的混合液在pH5.5的环境下荧光胺的荧光强度随时间的变化图;
图5为实施例5制备得到的载药纳米粒子对金黄色葡萄球菌生物膜(S.aureusbiofilm)中细菌的杀伤能力图。
具体实施方式
下面结合实施例和附图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。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
实施例中所用原料说明如下:N,N-二甲基丙烯酸胺(DMA),甲基丙烯酸酯(BMA),购自Sigma-Aldrich公司,2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl),1H-吡唑-1-甲脒盐酸盐,2,3-二甲基马来酸酐(DA)和N,N-二异丙基乙胺(DIPEA)购自安耐吉公司。偶氮二异丁腈(AIBN)购自Acros并用95%乙醇重结晶纯化。二氯甲烷,二甲基亚砜等试剂均购自国药集团化学试剂有限公司。采用由Milli-QSP超纯水制备系统(电阻率为18.4MΩ·cm)制备的超纯水。MH,LB的细菌培养基,琼脂购自广东环凯生物科技有限公司。
实施例1:聚合物亲水段P(DMA-co-AEMA)的制备
将小分子链转移剂4-氰基-4-(苯基硫代甲酰硫基)戊酸(CPADB)(12.7mg,0.045mmol)、二甲基丙烯酸胺(DMA,270mg,2.72mmol)、2-氨基乙基甲基丙烯酸酯盐酸盐(AEMA·HCl,300mg,1.81mmol)和偶氮二异丁腈(AIBN,1.5mg,0.01mmol)在安瓿瓶中溶于0.85mL二甲基亚砜。利用液氮迅速冷冻安瓿瓶中液体,后用油泵抽气,采取反复冻融三次的方法,使安瓿瓶中的反应环境没有空气和水分,达到真空的状态。然后在油泵抽气的状态下,利用酒精喷灯迅速将安瓿瓶密封。将密封好的安瓿瓶置于70℃油浴锅中搅拌反应15h,后用液氮终止聚合反应,打开安瓿瓶。利用乙醚对反应混合物进行沉降后离心,离心所得沉淀物再次溶解到少许二氯甲烷中并再次利用乙醚沉降,如此反复三次。最终得到红色固体,置于真空干燥箱中过夜干燥,最终得到产物。
具体反应路线如下:
实施例2:聚合物P(DMA-co-AEMA)-b-PBMA制备
P(DMA-co-AEMA)(128mg,0.012mmol),BMA(102.4mg,072mmol)和AIBN(0.4mg,0.0024mmol),溶解于1.6mL二甲基亚砜中,转移至安瓿瓶中。采取实施例1中的反复冻融三次的方法,移除混合物中的水和空气,真空条件下密封,70℃油浴锅中搅拌反应15h,后用液氮终止聚合反应,打开安瓿瓶,将反应后混合物转移至透析袋中,采用1000kDa的透析袋,常温透析过夜,冷冻干燥,得到白色粉末。
具体反应路线如下:
实施例3:聚合物P(DMA-co-GEMA)-b-PBMA制备
聚合物P(DMA-co-AEMA)-b-PBMA(134.5mg,0.009mmol),1H-吡唑-1-甲脒盐酸盐(75.2mg,0.513mmol)和DIPEA(104mg,1.026mmol)溶解于3.5mL的二甲基亚砜中,采取实施例1中的反复冻融三次的方法,移除混合物中的水和空气,真空条件下密封,55℃油浴锅中搅拌反应12h,后用液氮终止反应,打开安瓿瓶,将反应后混合物转移至1000kDa透析袋中,常温透析过夜,冷冻干燥,得到白色粉末。
具体反应路线如下:
实施例4:隐蔽胍基的聚合物P(DMA-co-GEMADA)-b-PBMA制备
聚合物P(DMA-co-GEMA)-b-PBMA(65mg,0.0042mmol),二甲基马来酸酐(DA,48.6mg,0.474mmol)和DIPEA(81.7mg,0.632mmol)溶解于0.7mL二氯甲烷和0.7mL二甲基亚砜中,常温搅拌反应24h后,将反应后混合物转移至透析袋1000KDa中,在弱碱性无菌水(pH8)中常温透析过夜,冷冻干燥,得到白色粉末。
具体反应路线如下:
实施例5:载药纳米粒子的构建及药物装载
将4mg P(DMA-co-GEMADA)-b-PBMA和4mg光热剂(长沙红)溶于二甲基亚砜。快速加入到9mL剧烈搅拌的去离子水,并继续搅拌30min。完成组装后的溶液转移至透析袋(3500KDa)中除去有机溶剂和未被装载的光热剂。每2h换一次水,透析12h。
对组装好的载药纳米粒子进行紫外吸收测定,结果如图1所示,可以看出纳米粒子中成功装载了光热试剂。对组装好的纳米粒子的尺寸进行相应的表征,结果如图2所示,可以看出纳米粒子的直径约为68nm,该纳米粒子的尺寸大小有利于渗透进入细菌生物膜。
实施例6:荧光胺检测胍基释放
取1mL聚合物P(DMA-co-GEMADA)-b-PBMA(170ug mL-1)水溶液并加入0.1mL荧光胺水溶液(1.5mg mL-1)。将混合物分别置于pH7.4和pH6的环境下孵育不同时间,利用MalvernZetasizer Nano ZS90 instrument(Malvern,UK)检测不同时间下荧光胺的荧光强度。结果分别如图3和图4所示,从图3中可以看出,混合物在pH7.4的环境下孵育,荧光胺的荧光强度不会随时间的变化而变强,证明该聚合物在pH7.4的环境下可以很好的屏蔽胍基。从图4中可以看出,混合物在pH5.5的环境下孵育,荧光胺的荧光强度会随时间的延长而变强,证明该聚合物在pH5.5的环境下,二甲基马来酸酐会逐渐脱落,暴露出胍基。
实施例7:梯度稀释涂布平板法测试纳米粒子对金黄色葡萄球菌生物膜(S.aureusbiofilm)中细菌的杀伤能力
该实验选择了金黄色葡萄球菌(ATCC 6538)作为革兰氏阳性菌的典型代表。首先,将保存在-20℃的S.aureus菌种接种到10mL的LB肉汤培养基(购买自广东环凯微生物科技有限公司)中,在37℃细菌培养摇床中培养过夜,后利用LB培养基将S.aureus悬浊液稀释至OD600为0.02。取200μL细菌悬浮液加入到96孔板中,培养24h后置换新鲜的LB培养基,共培养72h更换2次新鲜培养基。培养完成后,利用PBS清洗三次,洗去悬浮的金黄色葡萄球菌,取得含有成熟金黄色葡萄球菌生物膜的96孔板。
将实施例5中组装好的200μL载药纳米粒子水溶液(命名为响应型)(150μg/mL)加入到含有成熟金黄色葡萄球菌的孔中,与金黄色葡萄球菌生物膜共同孵育2h后,照光组采用730nm激光器照射5min。完成光照后立刻吸出多余药物,并用无菌水清洗掉多余药物。将整块96孔板密封好,放入超声仪中超声震荡10min,使得细菌生物膜中的细菌重新悬浮至溶液中。后梯度稀释,并进行平板涂布,统计每块平板的菌落数并进行分析。该试验组命名为响应型+光;
同时设置相应的对照组,如将200μL载药纳米粒子水溶液替换为等量的PBS缓冲液,且同样采用730nm激光器照射5min,将该对照组命名为:PBS+光;如将200μL载药纳米粒子替换为等量的PBS缓冲液,但不采用激光器照射,命名为PBS;如将200μL载药纳米粒子水溶液(150μg/mL)替换为等体积等浓度的非响应型载药纳米粒子水溶液,且同样采用730nm激光器照射5min,将该对照组命名为:非响应型+光;如将200μL载药纳米粒子水溶液(150μg/mL)替换为等体积等浓度的非响应型载药纳米粒子水溶液,但不采用激光器照射,命名为非响应型。其中非响应型载药纳米粒子水溶液由以下方法制备得到:将实施例4中的原料二甲基马来酸酐替换为等摩尔量的丁二酸酐,其余的操作均不变,得到不可以酸响应暴露胍基的聚合物,然后将该聚合物替换实施例5中的P(DMA-co-GEMADA)-b-PBMA,构建得到的浓度为纳米粒子浓度为150μg/mL的非响应型载药纳米粒子水溶液;
结果如图5所示,经过对平板上的菌落进行统计后得出结论,可以明显看出响应性的聚合物结合光热治疗能够更好的杀伤细菌生物膜内部的细菌。
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 (10)
1.一种酸响应电荷屏蔽聚合物,其特征在于由以下方法制备得到:
(1)将亲水单体和小分子链转移剂在引发剂存在条件下进行聚合,得到大分子链转移剂;;
(2)将大分子链转移剂和疏水单体在引发剂存在条件下进行聚合,得到聚合物1;
(3)在催化剂存在条件下,将聚合物1和1H-吡唑-1-甲脒盐酸盐反应,得到富含胍基的聚合物2;
(4)在催化剂存在条件下,将富含胍基的聚合物2与马来酸酐反应,得到胍基被屏蔽的聚合物,即酸响应电荷屏蔽聚合物。
2.根据权利要求1所述的酸响应电荷屏蔽聚合物,其特征在于:
步骤(1)中所述的亲水单体为氨基封端的亲水单体、N,N-二甲基丙烯酸胺、乙二醇、甲基丙烯酸二甲氨基乙酯中的至少一种;其中氨基封端的亲水单体为2-氨基乙基甲基丙烯酸酯盐酸盐、N-(3-氨基丙基)甲基丙烯酰胺盐酸盐、赖氨酸中的至少一种;
步骤(1)中所述的小分子链转移剂为4-氰基-4-(苯基硫代甲酰硫基)戊酸、二硫代氨基甲酸酯、三硫代碳酸酯中的至少一种。
3.根据权利要求1所述的酸响应电荷屏蔽聚合物,其特征在于:
步骤(1)中所述的亲水单体为2-氨基乙基甲基丙烯酸酯盐酸盐和N,N-二甲基丙烯酸胺的混合物;
步骤(1)中所述的小分子链转移剂为4-氰基-4-(苯基硫代甲酰硫基)戊酸;
步骤(1)中所述的引发剂为偶氮二异丁腈、偶氮二异庚腈、偶氮二异丁二甲酯中的一种;步骤(1)中聚合反应所用的溶剂为二甲基亚砜、二氯甲烷、1,4-二氧六环、无水乙醇和二甲基甲酰胺中的至少一种;
步骤(1)中所述的2-氨基乙基甲基丙烯酸酯盐酸盐、N,N-二甲基丙烯酸胺、小分子链转移剂以及引发剂的摩尔比为2~1200:2~1200:1~100:0.1~3;
步骤(1)中所述的聚合是指在真空密封条件下60~80℃反应12~75h。
4.根据权利要求1所述的酸响应电荷屏蔽聚合物,其特征在于:
步骤(2)中所述的疏水单体为甲基丙烯酸酯、苯乙烯,乙烯基三甲基硅烷等中的至少一种;步骤(2)中所述的引发剂为偶氮二异丁腈、偶氮二异庚腈、偶氮二异丁二甲酯中的一种。
步骤(2)中所述的大分子链转移剂、疏水单体和引发剂的摩尔比为1~100:2~1200:0.2~3;
步骤(2)中所述的聚合是指在真空条件下65~85℃反应12~24h,聚合反应的溶剂为二甲基亚砜,二氯甲烷、1,4二氧六环中的至少一种。
5.根据权利要求1所述的酸响应电荷屏蔽聚合物,其特征在于:
步骤(3)中所述的催化剂为N,N-二异丙基乙胺、三乙胺中的至少一种;
步骤(3)中所述的催化剂、聚合物1和1H-吡唑-1-甲脒盐酸盐的摩尔比为6~800:0.1~10:3~500;
步骤(3)中所述的反应是指在溶剂存在和真空的条件下20℃~75℃搅拌反应12~72h;反应的溶剂为二甲基亚砜。
6.根据权利要求1所述的酸响应电荷屏蔽聚合物,其特征在于:
步骤(4)中所述的催化剂为N,N-二异丙基乙胺;
步骤(4)中所述的催化剂、富含胍基的聚合物2和马来酸酐的摩尔比为6~800:0.1~1:3~500;
步骤(4)中所述的反应是指在溶剂存在条件下常温反应12~72h,其中溶剂为二甲基亚砜、二氯甲烷中的至少一种。
7.根据权利要求1~6任一项所述的酸响应电荷屏蔽聚合物作为载体在制备载药纳米粒子中的应用。
8.一种载药纳米粒子,其特征在于由以下方法制备得到:
将具有抗菌活性的物质与权利要求1~6任一项所述的酸响应电荷屏蔽的聚合物共同溶解在有机溶剂中,然后加入到正在搅拌的水中,继续搅拌0.5~12h,再通过透析或者超滤法除去有机溶剂,最终得到载药纳米粒子;
或者,将具有抗菌活性的物质与权利要求1~6任一项所述的酸响应电荷屏蔽的聚合物共同溶解在有机溶剂中,在搅拌的条件下向其中缓慢注入水,注水速度为每小时0.09~9mL,注水完毕后再通过透析或者超滤法去除有机溶剂,最终得到载药纳米粒子。
9.根据权利要求8所述的载药纳米粒子,其特征在于:
所述的光敏剂为吲哚菁绿、曙红Y、盐酸氨酮戊酸中的至少一种,所述的光热试剂为长沙红、纳米金棒、聚吡咯中的至少一种,所述的抗生素为阿莫西林、头孢哌酮、红霉素、庆大霉素中的至少一种;
所述的有机溶剂为1,4-二氧六环、二甲基亚砜、二甲基甲酰胺中的至少一种。
10.根据权利要求8所述的载药纳米粒子,其特征在于:
所述的具有抗菌活性的物质与酸响应电荷屏蔽聚合物的质量比为1:1~100;所述的有机溶剂与水的体积比为1:9;所述的酸响应电荷屏蔽聚合物与水的质量体积比为0.1~100mg/L;
所述的透析是利用分子量为1kDa以上的透析膜在去离子水中进行透析;所述的超滤法是指用使用分子量10kDa以上的超滤管,通过离心去除有机溶剂。
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