CN115555576A - 一种等离子共振表面多层自组装小分子结构、组装方法及其应用 - Google Patents
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Abstract
本发明属于生物医药的技术领域,具体涉及一种等离子共振表面多层自组装小分子结构、组装方法及其应用,通过分析具有自组装性质的小分子进行结构分析,可以实现反向的结构设计:根据所需要的功能选择小分子类型;然后对小分子顶端功能团用氨基或胺基修饰;侧链端基用氨基、胺基、醚基、羧酸基、酯基或醛基等富O、N、P、S进行修饰;修饰过后的小分子配置成0.1‑5.0μmol/L的水溶液;即可加入新和成的银纳米颗粒进行自组装,制备得到多层小分子自组装的银纳米颗粒结构。所述的多层自组装小分子结构可用于分子检测以及杀菌。
Description
技术领域
本发明属于生物医药的技术领域,具体涉及一种等离子共振表面多层自组装小分子结构、组装方法及其应用。
背景技术
活性金属纳米颗粒表面等离子共振特性,表面十分活跃,这种性质是一柄双刃剑,一方面形成了有很多可以利用的独特性质,比如表面增强拉曼光谱,银光猝灭等光学性质用于传感和检测;另一方面,活跃的表面极不稳定,容易聚集,也容易吸附和影响周围的分子结构。以银纳米颗粒为例,在杀菌、光学等方面有很大应用前景,但是极不稳定,必须有表面涂层作为稳定剂。因此,制备结构均一稳定、性质可控的银纳米颗粒一直是科研的重点,也是应用的瓶颈。
发明内容
为了克服现有技术中的不足,本发明提供了一种等离子共振表面多层自组装小分子结构、组装方法及其应用的解决方案,具体如下:
一种等离子共振表面多层自组装小分子的组装方法,包括如下步骤:
S1.小分子主结构选择:根据实际需要的小分子功能确定小分子框架结构;
S2.小分子修饰:小分子顶端功能团用氨基或胺基修饰;侧链端基用氨基、胺基、醚基、羧酸基、酯基或醛基富O、N、P、S进行修饰;在化学修饰之前,设计的结构可以用高斯软件进行模拟,确定分子之间氢键相互作用力以及自组装排列可行性,然后进行实验室合成,节省了工作时间和成本。
S3.将合成的小分子配置成0.1-5.0μmol/L的水溶液,备用;
S4.合成银纳米颗粒:用柠檬酸钠在加热条件下还原硝酸银,制备银纳米颗粒,经过两次离心洗涤,配置成100μmol/L的的水溶液,备用;
S5.自组装结构形成:将银纳米溶液稀释至浓度为0.3-25μmol/L的水溶液,将小分子和银纳米混合搅拌10分钟,即可得到多层小分子自组装包裹的银纳米结构溶液,该溶液可通过离心-再分散的过程进行浓缩,增加自组装银纳米结构浓度。
作为一种等离子共振表面多层自组装小分子的组装方法的优选技术方案,用于多层自组装的小分子框架通常含有一条或多条支链,且支链长度不长于主链的1/2,支链太长的话,影响分子间氢键的形成,会使分子的有序排列受到限制,分子在同一层的自组装受阻,或者层与层之间的排列受阻,使得“多层自组装”无法实现。
作为一种等离子共振表面多层自组装小分子的组装方法的优选技术方案,所述小分子包括罗丹明6G和四环素。
作为一种等离子共振表面多层自组装小分子的组装方法的优选技术方案,小分子:银纳米的体积比例为1:1-1:5。
作为一种等离子共振表面多层自组装小分子的组装方法的优选技术方案,其特征在于,自组装银纳米结构浓度不超过60μmol/L,浓度超过60umol/L时,自组装结构会出现融合,随着浓度继续升高,回出现团聚,进而自组装结构破坏。
一种多层自组装小分子结构,由上述的组装方法制备而成,多层自组装小分子结构是一种连续多层的自组装结构,且其高达36层。
自组装结构基于两种或三种化学键(电荷+范德华力或氢键):第一层小分子与银表面的阴阳离子电荷相互作用,第二层以上的多层小分子之间或者同一层的小分子之间以范德华力或氢键相互作用,其中氢键形成的自组装结构更为稳定。同一层的小分子之间和不同层的小分子之间都可以是范德华力或者氢键。通常以氢键为主,更稳定。
制备而成的多层自组装小分子结构可用于分子检测以及杀菌。
本发明的有益效果为:
1.自组装结构基于两种或三种化学键(电荷+范德华力或氢键):第一层小分子与银表面的阴阳离子电荷相互作用,第二层以上的多层小分子之间或者同一层的小分子之间以范德华力或氢键相互作用,其中氢键形成的自组装结构更为稳定;
2.首先,根据荧光猝灭的量,可以用于检测溶液中存在的罗丹明分子浓度;同时,荧光猝灭现象本身可以用于荧光分子浓度的检测;银光猝灭之后的自组装银纳米结构,在与其他物质相互作用发生反应,结构被打破,罗丹明分子释放出来,会恢复荧光。根据这些荧光恢复的量,可以用于判定溶液中可能存在的强氧化性物质以及其浓度。
3.四环素自组装结构相对纯的四环素分子和纯的银纳米颗粒,具有协同的杀菌效果。
附图说明
图1是罗丹明6G化学分子结构图。
图2是实施例一中荧光猝灭图谱;
图3是罗丹明分子围绕一个银纳米颗粒自组装形成的动态结构图;
图4是四环素分子化学结构图;
图5是不同浓度的四环素自组装结构对耐药性沙门氏细菌DT104的抑制效果;
图6是四环素自组装结构杀菌机理过程图;
具体实施方式
下面将结合附图和实施例对本发明做进一步地说明。
实施例一
罗丹明6G是一种荧光燃料,其化学分子结构如图1所示,根据其理化性质和其结构,制备出罗丹明6G在银纳米颗粒表面自组装结构,用于光学检测,为实现这一目的,其制备步骤包括如下:
在化学修饰之前,设计的结构可以用高斯软件进行模拟,确定分子之间氢键相互作用力以及自组装排列可行性,然后进行实验室合成;将合成的罗丹明6G小分子配置成1μmol/L的水溶液,备用;用柠檬酸钠在加热条件下还原硝酸银,制备银纳米颗粒,经过两次离心洗涤,配置成100μmol/L的的水溶液,备用;将700μL浓度为100μmol/L的银纳米颗粒逐渐加入1000μL浓度为1μmol/L的罗丹明6G溶液中,混合搅拌10分钟,自组装结构逐渐形成,即可得到多层小分子自组装包裹的银纳米结构溶液,银纳米颗粒按照荧光淬灭滴定法逐渐加入到罗丹明6G溶液中,通过荧光光谱仪可以获得荧光猝灭图谱,如图2所示。
1μmol/L的罗丹明6G溶液在逐渐加入银纳米颗粒时,发生自组装,结果就会导致荧光强度降低,造成了荧光的猝灭。当荧光强度趋于稳定时,罗丹明自组装的银纳米结构浓度达到饱和。其中横坐标是波长,纳米;纵坐标为荧光强度;从上到下是逐渐加入银纳米颗粒时罗丹明6G溶液的荧光强度变化。首先,根据荧光猝灭的量,可以用于检测溶液中存在的罗丹明分子浓度;同时,荧光猝灭现象本身可以用于荧光分子浓度的检测;银光猝灭之后的自组装银纳米结构,在与其他物质相互作用发生反应,结构被打破,罗丹明分子释放出来,会恢复荧光。根据这些荧光恢复的量,可以用于判定溶液中可能存在的强氧化性物质以及其浓度。
在溶液中形成的动态结构,如图3所示。
该结构表示了多层(最高可达36层)罗丹明分子围绕一个银纳米颗粒自组装形成的动态结构图。Cl-表示氯离子,R6G表示罗丹明分子。该实验中采用了吕罗丹明氯盐。
实施例二
四环素是广谱抗生素,对多数革兰阳性与阴性菌有抑制作用,高浓度有杀菌作用,并能抑制立克次体、沙眼病毒等,对革兰阴性杆菌作用较好。其作用机制主要是阻止氨酰基与核糖核蛋白体的结合,阻止肽链的增长和蛋白质的合成,从而抑制细菌的生长,高浓度时也有杀菌作用,其分子化学结构如图4所示。通过自组装结构,利用四环素和银纳米颗粒的协同杀菌作用,制备针对耐药性细菌的高效广谱抗生素,四环素分子在银纳米表面自组装结构制备步骤:在化学修饰之前,设计的结构可以用高斯软件进行模拟,确定分子之间氢键相互作用力以及自组装排列可行性,然后进行实验室合成;将合成的四环素小分子配置成5μmol/L的水溶液,备用;用柠檬酸钠在加热条件下还原硝酸银,制备银纳米颗粒,经过两次离心洗涤,配置100μmol/L的水溶液,备用;将银纳米溶液稀释至浓度为20μmol/L的水溶液,将四环素小分子和银纳米按照体积比1:5混合搅拌10分钟,即可得到多层小分子自组装包裹的银纳米结构溶液,该溶液可通过离心-再分散的过程进行浓缩,增加自组装银纳米结构浓度,测试不同浓度(0.5μmol/L、2.0μmol/L、8.0μmol/L以及16.0μmol/L)的自组装银纳米结构对耐药性沙门氏菌DT104的杀菌效果。
实施例三
测试不同浓度(0.5μmol/L、2.0μmol/L、8.0μmol/L以及16.0μmol/L)的四环素分子对耐药性沙门氏菌DT104的杀菌效果。
对照例一
测试银离子对耐药性沙门氏菌DT104的杀菌效果。
对照例二
测试银纳米颗粒对耐药性沙门氏菌DT104的杀菌效果。
四环素自组装结构相对纯的四环素分子和纯的银纳米颗粒,具有协同的杀菌效果。不同浓度的自组装结构对耐药性沙门氏细菌DT104的抑制效果如图5所示,浓度根据自组装结构中抗生素分子结构计算:
横坐标为对照组及实验组的浓度,纵坐标为沙门氏菌被抑制的百分比。纯的银纳米颗粒对沙门氏菌抑制率约在15%,0.5–16μM的四环素对沙门氏菌抑制率约在25-30%,而同等浓度形成的自组装结构对沙门氏菌的抑制均超过了98%。证明了自组装结构实现并加强了四环素和银纳米颗粒的协同杀耐药性细菌效果。
杀菌机理过程图,如图6所示,在整个协同杀菌过程中,四环素首先与因奶相互作用,在银纳米颗粒表面自组装,然后自组装结构以一个整体结构与细菌细胞相互识别,识别过程中相互作用,银纳米胶束结构释放银离子和抗生素小分子,形成局部过量的抗生素小分子以及银离子,银离子可直接游离于细胞内部,与染色体、核糖体等细胞器或细胞结构非特异性结合。这种结合可以造成两种效果,一是导致细菌细胞正常功能丧失,直接导致细胞衰亡,二是导致细菌细胞对传统抗生素分子耐药机理的信号通路中断,使得细胞对于传统抗生素分子的进攻无法做出耐受性的反应。这种耐药性通路的丧失配合局部高浓度的传统抗生素分子,可以快速高效的彻底瓦解耐药性细菌。
以上以用实施例说明的方式对本发明作了描述,本领域的技术人员应当理解,本公开不限于以上描述的实施例,在不偏离本发明的范围的情况下,可以做出各种变化、改变和替换。
Claims (9)
1.一种等离子共振表面多层自组装小分子的组装方法,其特征在于,包括如下步骤:
S1.小分子主结构选择:根据实际需要的小分子功能确定小分子框架结构;
S2.小分子修饰:小分子顶端功能团用氨基或胺基修饰;侧链端基用氨基、胺基、醚基、羧酸基、酯基或醛基富O、N、P、S进行修饰;
S3.将合成的小分子配置成0.1-5.0μmol/L的水溶液,备用;
S4.合成银纳米颗粒:用柠檬酸钠在加热条件下还原硝酸银,制备银纳米颗粒,经过两次离心洗涤,配置成100μmol/L的的水溶液,备用;
S5.自组装结构形成:将银纳米溶液稀释至浓度为0.3-25μmol/L的水溶液,将小分子和银纳米混合搅拌10分钟,即可得到多层小分子自组装包裹的银纳米结构溶液,该溶液可通过离心-再分散的过程进行浓缩,增加自组装银纳米结构浓度。
2.根据权利要求1所述的等离子共振表面多层自组装小分子的组装方法,其特征在于,用于多层自组装的小分子框架通常含有一条或多条支链,且支链长度不长于主链的1/2。
3.根据权利要求1所述的等离子共振表面多层自组装小分子的组装方法,其特征在于,所述小分子包括罗丹明6G和四环素。
4.根据权利要求1所述的等离子共振表面多层自组装小分子的组装方法,其特征在于,小分子:银纳米的体积比例为1:1-1:5。
5.根据权利要求1所述的等离子共振表面多层自组装小分子的组装方法,其特征在于,自组装银纳米结构浓度不超过60μmol/L。
6.一种多层自组装小分子结构,其特征在于,基于权利要求1至5任一所述的组装方法制备而成。
7.根据权利要求6所述的多层自组装小分子结构,其特征在于,多层自组装小分子结构是一种连续多层的自组装结构,且其高达36层。
8.根据权利要求7所述的多层自组装小分子结构,其特征在于,第一层小分子与银表面的阴阳离子电荷相互作用,第二层以上的多层小分子之间或者同一层的小分子之间以范德华力或氢键相互作用。
9.基于权利要求6至8任一所述的多层自组装小分子结构的应用,其特征在于,所述的多层自组装小分子结构可用于分子检测以及杀菌。
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