CN114539299A - 用于检测过氧亚硝酸阴离子的化合物和包括其的纳米颗粒及其应用 - Google Patents
用于检测过氧亚硝酸阴离子的化合物和包括其的纳米颗粒及其应用 Download PDFInfo
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- CN114539299A CN114539299A CN202111349938.0A CN202111349938A CN114539299A CN 114539299 A CN114539299 A CN 114539299A CN 202111349938 A CN202111349938 A CN 202111349938A CN 114539299 A CN114539299 A CN 114539299A
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Abstract
本发明属于生化检测领域,公开了一种用于检测过氧亚硝酸阴离子的化合物和包括其的纳米颗粒及其应用。本发明的化合物具有式(I)的结构。本发明的化合物可在细胞中和在生物体水平上进行选择性高灵敏度检测,可以用于制备炎症部位可视化的聚集诱导发光材料。
Description
技术领域
本发明属于生化检测领域,更具体而言,本发明涉及用于检测过氧亚硝酸阴离子的化合物和包括其的纳米颗粒及其应用。
背景技术
活性氧氮类物质(RONS)是一类重要的化学反应介质,包括活性氧和活性氮。RONS与多种生物过程密切相关,如氧化应激、细胞信号、心血管疾病等。其中,过氧亚硝酸阴离子(ONOO-)是一种重要的生物氧化剂,有助于调节氧化还原的稳态、激活信号转导和诱导正常的免疫反应。然而,过度的ONOO-常伴有炎症和神经退行性疾病、类风湿性关节炎等疾病的发生。
基于化学发光、荧光和电化学发光的RONS检测方法已有很多报道。在这些方法中,荧光探针以其高灵敏度、无创性、实时检测等优点受到越来越多的关注。尽管如此,现在也只有一小部分的荧光检测策略可以实现对ONOO-的特异性检测,而且能同时实现在生物体内和体外检测的更少。另外,传统的荧光探针如罗丹明、荧光素、甲酚紫等在稀溶液中通常会发出明亮的光,但在聚集状态会产生聚集诱导猝灭(Aggregation-caused quenching,ACQ)的效应。而且,这些探针的光稳定性差、斯托克斯位移短、信噪比低,这些都限制了它们的应用。因此,本领域中需要对RONS,特别是过氧亚硝酸阴离子(ONOO-)进行检测的方法。
发明内容
本发明的目的至少在于提供一种对ONOO-具有选择性高灵敏度检测效果的化合物。
因此,在本发明的第一方面中,本发明提供了一种化合物,所述化合物具有式(I)的结构:
其中,抗衡阴离子X-为具有一个或多个电荷的阴离子,如选自:F-、Cl-、Br-、I-、At-、Ts-、PF6 -、BF4 -、SbF6 -、SbF5 -、CH3COO-、CF3COO-、CO3 2-、SO4 2-、SO3 2-、CF3SO2 -、TsO-、ClO4 -、(F3CSO2)N-和PO4 3-;
每个R、R1、R2、R3和R4可独立地选自:氢、氟、氯、溴、碘、烷基、烷氧基、羟烷基、氨基、烷氨基、烷基、不饱和烷基、杂烷基、环烷基、杂环烷基、芳基和杂芳基中的一种或几种。
在一个实施方案中,所述化合物具有式(II)的结构:
在本发明的第二方面中,本发明提供了包括本发明第一方面的化合物的纳米颗粒。
在一个实施方案中,所述纳米颗粒以二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)为包封基质。
在一个实施方案中,所述纳米颗粒的流体动力学直径约为150纳米。
在本发明第三方面中,本发明提供了本发明第一方面的化合物和本发明第二方面的纳米颗粒用于检测过氧亚硝酸阴离子的应用。
在一个实施方案中,所述化合物与非其他活性氮氧类物质无反应,所述其他活性氮氧类物质优选包括ClO-、H2O2、·OH、tBuOOH、·OBu、O2 -、1O2、NO2 -和NO3 -。
在一实施例中,所述检测过氧亚硝酸阴离子包括检测所述化合物与过氧亚硝酸阴离子反应后的产物发射的荧光。
在一实施例中,所述检测过氧亚硝酸阴离子包括在细胞水平检测活性氮氧类物质。
在一实施例中,所述检测过氧亚硝酸阴离子包括在pH大于6.8,优选pH6.8-pH7.4,更优选pH7.4,下检测过氧亚硝酸阴离子。
在一实施例中,所述应用包括对炎症相关的吞噬细胞进行染色。
在一实施例中,所述应用包括对炎症部位进行可视化鉴定。
本发明的化合物可在细胞中和生物体水平上进行选择性高灵敏度检测,可以用于制备炎症部位可视化的聚集诱导发光材料。
附图说明
图1示出了本发明的AIE荧光探针TPE-DMAB的合成路线(a);在ONOO-存在下TPE-DMAB向TPE-DMA转化的可能机理(b)。
图2示出了本发明的AIE荧光探针TPE-DMAB及其衍生物的结构示意图。
图3示出了TPE-DMAB的1H核磁谱(CD3OD,400MHz,298K)。
图4示出了TPE-DMAB的13C核磁谱(CD3OD,400MHz,298K)。
图5示出了TPE-DMAB的质谱。
图6示出了TPE-DMA(10-5M)在四氢呋喃(THF)中的吸收光谱。
图7示出了不同含水率(fW)的THF/水混合物中TPE-DMA(10-5M)的PL光谱,激发波长:360nm。
图8示出了THF/水混合物中I/I0与水分数的关系图,I0和I分别是TPE-DMA(10-5M)在THF和THF/水混合物中的PL强度。
图9示出了TPE-DMAB(10-5M)在DMSO中的吸收光谱。
图10示出了不同ONOO-浓度孵育的TPE-DMAB的荧光光谱(a);I/I0与ONOO-的浓度对比图(b),I0和I是在530纳米处TPE-DMAB在加入ONOO-前后的荧光强度,插图示出ONOO-检测的线性范围;
TPE-DMAB荧光活化动力学研究(c);在PBS缓冲液中I/I0与多种活性氧氮类物质(RONS)的关系曲线(d),I0和I是在530纳米处TPE-DMAB在加入RONS前后的荧光强度。
图11示出了pH对TPE-DMAB的影响。
图12示出了TPE-DMAB纳米粒子对ONOO-的荧光响应示意图(a);TPE-DMAB纳米粒子的DLS粒径图,插图示出TPE-DMAB纳米粒子的TEM图像(b);不同浓度ONOO-对TPE-DMAB纳米粒子的荧光响应(c),插图是在PBS缓冲液中加入200微摩尔TPE-DMAB纳米粒子前后的荧光图像。
图13示出了TPE-DMAB在PBS缓冲液(pH 7.4)中有无ONOO-孵育的吸收光谱。
图14示出了磷酸盐缓冲生理盐水(PBS)缓冲液(pH=7.4)中I/I0与各种活性氧氮类物质(RONS)的柱状图,I和I0分别是TPE-DMAB纳米粒子在存在和不存在RONS时的PL强度。
图15示出了酸碱度(pH)在ONOO-存在下对TPE-DMAB纳米粒子的影响。
图16示出了用TPE-DMAB纳米粒子(3μg·mL-1)孵育3小时的活小鼠巨噬细胞(RAW264.7)的CLSM图像:对照组为未经处理的细胞(a);LPS(2h)和PMA(0.5h)处理的细胞(b);细胞在用LPS(2h)和PMA(0.5h)处理前用NAC处理的细胞(c)。比例尺:20微米。
图17示出了细胞毒性实验:MTT法检测TPE-DMAB纳米粒子对4T1细胞的杀伤作用。
图18示出了原位注射TPE-DMAB纳米粒子30分钟前(a)、后(b),以及尿酸预处理(c)的裸鼠体内LPS诱导炎症的荧光图像,蓝色圆圈表示小鼠左背部的炎症区域。
图19示出了注射TPE-DMAB纳米粒子的小鼠感染皮肤中的定量平均荧光(n=3)。
具体实施方式
下面详细描述本发明的实施方案。下面描述的实施方案是示例性的,仅用于解释本发明,而不能理解为对本发明的限制。实施方案中未注明具体技术或条件的,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
定义和一般术语:
现在详细描述本发明的某些实施方案,其实例由随附的结构式和化学式说明。本发明意图涵盖所有的替代、修改和等同技术方案,它们均包括在如权利要求定义的本发明范围内。本领域技术人员应认识到,许多与本文所述类似或等同的方法和材料能够用于实践本发明。本发明绝不限于本文所述的方法和材料。在所结合的文献、专利和类似材料的一篇或多篇与本申请不同或相矛盾的情况下(包括但不限于所定义的术语、术语应用、所描述的技术,等等),以本申请为准。
应进一步认识到,本发明的某些特征,为清楚可见,在多个独立的实施方案中进行了描述,但也可以在单个实施例中以组合形式提供。反之,本发明的各种特征,为简洁起见,在单个实施方案中进行了描述,但也可以单独或以任意适合的子组合提供。
除非另外说明,本发明所使用的所有科技术语具有与本发明所属领域技术人员的通常理解相同的含义。本发明涉及的所有专利和公开出版物通过引用方式整体并入本发明。
除非另外说明,应当应用本文所使用的下列定义。出于本发明的目的,化学元素与元素周期表CAS版,和《化学和物理手册》,第75版,1994一致。此外,有机化学一般原理可参考“Organic Chemistry”,Thomas Sorrell,University Science Books,Sausalito:1999,和“March's Advanced Organic Chemistry”by Michael B.Smith and Jerry March,JohnWiley&Sons,New York:2007中的描述,其全部内容通过引用并入本文。
除非另有说明或者上下文中有明显的冲突,本文所使用的冠词“一”、“一个(种)”和“所述”旨在包括“至少一个”或“一个或多个”。因此,本文所使用的这些冠词是指一个或多于一个(即至少一个)宾语的冠词。例如,“一组分”指一个或多个组分,即可能有多于一个的组分被考虑在所述实施方案的实施方式中采用或使用。
术语“包含”为开放式表达,即包括本发明所指明的内容,但并不排除其他方面的内容。
另外,需要说明的是,除非以其他方式明确指出,在本发明中所采用的描述方式“各…独立地为”与“…各自独立地为”和“…独立地为”可以互换,均应做广义理解,其既可以是指在不同基团中,相同符号之间所表达的具体选项之间互相不影响,也可以表示在相同的基团中,相同符号之间所表达的具体选项之间互相不影响。
在本说明书的各部分,本发明公开化合物的取代基按照基团种类或范围公开。特别指出,本发明包括这些基团种类和范围的各个成员的每一个独立的次级组合。例如,术语“C1-18烷基”包括甲基、乙基、C3烷基、C4烷基、C5烷基和C6烷基。
在本发明的各部分,描述了诸如L等连接取代基。当该结构清楚地需要连接基团时,针对该基团所列举的马库什变量应理解为连接基团。例如,如果该结构需要连接基团并且针对该变量的马库什基团定义列举了“烷基”或“芳香族基团”,则应该理解,该“烷基”或“芳基”分别代表连接的亚烷基基团或亚芳香族基团。
本发明使用的术语“烃基”包括芳香族烃基和脂肪族烃基。脂肪族烃基包括“烷基”或“烷基基团”,烯基和炔基,它们可以是饱和或不饱和的直链或支链二价烃基基团。所述烃基可以任选地被一个或多个本发明描述的取代基所取代。在本发明的一个实施方案中,烷基基团含有1-18个碳原子。在另一实施方案中,烷基基团含有1-12个碳原子;在又一实施方案中,烷基基团含有1-6个碳原子;再一实施方案中,烷基基团含有1-4个碳原子;还在一实施方案中,烷基基团含有1-3个碳原子。
烷基基团的实例包含,但并不限于,C1-12烷基,如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、2-戊基、3-戊基、2-甲基-2-丁基、3-甲基-2-丁基、3-甲基-1-丁基、2-甲基-1-丁基、正己基、2-己基、3-己基、2-甲基-2-戊基、3-甲基-2-戊基、4-甲基-2-戊基、3-甲基-3-戊基、2-甲基-3-戊基、2,3-二甲基-2-丁基、3,3-二甲基-2-丁基、正庚基、正辛基等等。
术语“烯基”表示碳原子的直链或支链一价烃基,其中至少有一个不饱和位点,即有一个碳-碳sp2双键,其中,所述烯基基团任选地被一个或多个本发明所描述的取代基所取代,其包括“cis”和“tans”的定位,或者"E"和"Z"的定位。在一实施方案中,烯基基团包含2-8个碳原子;在另一实施方案中,烯基基团包含2-6个碳原子;在又一实施方案中,烯基基团包含2-4个碳原子。烯基基团的实例包括,但并不限于,乙烯基、烯丙基等等。
术语“炔基”表示碳原子的直链或支链一价烃基,其中至少有一个不饱和位点,即有一个碳-碳sp三键,其中,所述炔基基团任选地被一个或多个本发明所描述的取代基所取代。在一实施方案中,炔基基团包含2-8个碳原子;在另一实施方案中,炔基基团包含2-6个碳原子;在又一实施方案中,炔基基团包含2-4个碳原子。炔基基团的实例包括,但并不限于,乙炔基、炔丙基、1-丙炔基等等。
术语“羧基”,无论是单独使用还是和其他术语连用,如“羧烷基”,表示-CO2H;术语“羰基”,无论是单独使用还是和其他术语连用,如“氨基羰基”或“酰氧基”,表示-(C=O)-。
术语“卤素”和“卤代”是指氟(F)、氯(Cl)、溴(Br)或碘(I)。
术语“芳香族基团”包括从芳香环中去掉两个氢原子从而与其他基团直接连接的基团。优选地,芳香族基团在成环原子中具有至少一个杂原子,例如N、O或S。
术语“芳香族环烃基”包括单环、双环和三环的芳基,其中,至少一个环体系是芳香族的,其中每一个环体系包含6-18个原子组成的环。芳基基团通常,但不必须,通过芳基基团的芳香性环与母体分子连接。术语“芳基”可以和术语“芳香环”或“芳环”交换使用。芳基基团的实例可以包括苯基、联苯基、萘基和蒽。所述芳基基团任选地被一个或多个本文所描述的取代基所取代。
在本发明中,取代基可以选自卤原子、羟基、醛基、羧基、氨基、任选被一个或多个C6-C18芳香族环烃基或成环碳原子5-18的芳香族杂环基取代的C2-C18烯基、任选被一个或多个C6-C18芳香族环烃基或成环碳原子5-18的芳香族杂环基取代的C2-C18炔基、任选被一个或多个C6-C18芳香族环烃基或成环碳原子5-18的芳香族杂环基取代的的C1-C18烷基、成环碳原子6-18的芳香族环烃基、成环碳原子5-18的芳香族杂环基、巯基、氰基和硝基中的至少一种。
芳香族环烃基和芳香族杂环基的例子包括(例如)苯基、萘基、苯基、萘基、蒽基、菲基、并四苯基、芘基、苯并[c]菲基、苯并菲基、芴基、苯并芴基、二苯并芴基、联苯基、三联苯基、四联苯基、荧蒽基、吡咯基、吡嗪基、吡啶基、嘧啶基、三嗪基、吲哚基、异吲哚基、咪唑基、呋喃基、苯并呋喃基、异苯并呋喃基、二苯并呋喃基、二苯并噻吩基、喹啉基、异喹啉基、喹喔啉基、咔唑基、菲啶基、吖啶基、菲咯啉基、吩嗪基、吩噻嗪基、吩噁嗪基、噁唑基、噁二唑基、呋咱基、噻吩基、苯并噻吩基、二氢吖啶基、氮杂咔唑基、喹唑啉基等。
取代基的例子包括以下结构中的一种或多种:
在本发明中,发明人开发了一种荧光开启型生物探针TPE-DMAB(4,4'-(1,2-二苯基乙烯-1,2-二苯基)双(N-(4-硼苯基)-N,N-二甲基苯胺),用于ONOO-的特异性检测。TPE-DMAB可以作为一种新型离子型活性荧光生物探针,是一种由(4-(溴甲基)苯基)硼酸修饰的具有聚集诱导发光(Aggregation-induced emission,AIE)活性的探针。TPE-DMAB具有一定的亲水性,在水中表现出微弱的荧光发射,OONO-的存在会使探针氧化裂解,并最终产生疏水性的AIE分子,从而在水中形成聚集体并展现出明亮的荧光。该荧光探针具有聚集诱导发光(AIE)单元如四苯基乙烯(TPE),水溶性单元如季铵盐和硼酸,以及特异性识别过氧亚硝酸阴离子的基元如苯硼酸。
发明人发现,如上所述的AIE荧光探针在与过氧亚硝酸阴离子反应后的产物在生理环境会有明亮的荧光发射,从而实现所述化合物对过氧亚硝酸阴离子的可视化鉴定。其中所述荧光探针在生理(水)环境中无或只能发出微弱的荧光,但在过氧亚硝酸阴离子存在的环境中会发出530纳米左右的强烈荧光。发明人发现,该AIE探针显示出良好的生物相容性、光稳定性和制备简单等优点。TPE-DMAB可快速并高度选择性的对ONOO-进行反应,TPE-DMAB对ONOO-具有良好的灵敏度(检测限为54nM)。因此,TPE-DMAB(及其衍生物)可实现对过氧亚硝酸阴离子的特异性检测,且受其他RONS干扰小。进一步证实本探针可作为荧光成像工具监测细胞内过氧亚硝酸阴离子的水平和实现体内炎症的可视化。
TPE-DMAB可以被进一步制备成纳米粒子,所获得的纳米探针对ONOO-也表现出很高的选择性和特异性,制备的TPE-DMAB纳米粒子在病理水平上表现出极好的对ONOO-的选择性检测,这使得该生物探针能够实现ONOO-的体外检测(例如检测细胞中ONOO-),以及实现生物体内炎症部位的可视化。
实施例。
在实施例中,本发明的TPE-DMAB通过简便的合成路线被制备(图1)。本发明的AIE荧光探针TPE-DMAB及其衍生物如图2所示。在氮气保护下向烧瓶中添加100毫克(0.24mmol)TPE-DMA和120毫克(0.56mmol)4-(溴甲基)苯基硼酸;然后加入20毫升乙腈;反应在室温下搅拌30小时;然后通过旋转蒸发除去溶剂,再用过量的乙酸乙酯洗涤后收集产物并干燥为白色固体,产率为23%。1H NMR(400MHz,CD3OD)δ(ppm):8.19(s,4H),7.71(d,J=8.1Hz,4H),7.57(d,J=8.2Hz,4H),7.26(t,J=7.3Hz,4H),7.20(d,J=7.1Hz,2H),7.12(d,J=8.9Hz,4H),7.02(d,J=7.0Hz,4H),6.78(d,J=8.2Hz,4H),4.90(s,4H),3.49(s,12H).13C NMR(400MHz,CD3OD)δ(ppm):145.1,142.8,142.4,140.5,136.9,134.6,132.2,131.7,131.0,129.9,128.7,127.9,122.0,72.6,53.1.HRMS:m/z计算值为[M-H]+C44H45B2N2O4 +687.3560;实测值687.3579。核磁及质谱的表征如图3-图5所示。
本发明人对TPE-DMA的吸收光谱和荧光光谱进行了测试。TPE-DMA在四氢呋喃(THF)中在365nm处显示出强吸收峰(图6),并且在具有不同水比例的THF/水混合物中研究了其AIE特性(图7、图8)。当水含量小于70%时,TPE-DMA在溶液中几乎没有荧光,当水含量增加到90%时,TPE-DMA的荧光强度急剧增加,在530nm处出现明显的发射峰,这表明TPE-DMA有典型的AIE特性。同时,在二甲基亚砜(DMSO)中测量了TPE-DMAB的吸收光谱,发现吸收在280-440纳米范围内,最大值在315纳米处(图9)。
图10中a显示了TPE-DMAB与不同浓度OONO-反应前后的PL光谱。在DMSO/磷酸盐缓冲生理盐水(Phosphate buffered saline,PBS)(1:99,体积比)介质中,TPE-DMAB在没有OONO-的情况下几乎不发光;然而,在加入OONO-后,光谱在530纳米处出现明显的发射峰,随着OONO-的增加,荧光强度逐渐增强,如图10中b所示。当OONO-浓度达到20微摩尔每升时,发光强度提高了约100倍。此外,如图10中b的插图所示,相对发光强度(I/I0)与OONO-浓度在浓度范围3-12微摩尔每升内呈良好的线性关系。由此计算得知,OONO-的检出限约是54nM。
如图10中c所示,通过对TPE-DMAB动力学行为的研究发现,随着在TPE-DMAB溶液中加入OONO-,随着反应时间延长,荧光强度急剧增加,并在8分钟内达到最大值。而且,高浓度的OONO-导致的荧光强度的增强更快且能达到更大的增强倍数。而未加入OONO-的TPE-DMAB溶液在同一时间内保持不变,说明所述探针具有良好的稳定性。
酸碱度(pH)对检测系统的影响(图11)结果表明,该探针在pH7.4时荧光增强最大,表明该探针在生理环境中的性能最好。酸性条件不利于苯硼酸酯的裂解,导致荧光在酸性条件下不能被激活。
TPE-DMAB对不同RONS的反应结果表明。只有OONO-可显著触发荧光增强,而其他RONS对所述探针显示可忽略的响应(如图10中d所示),这表明TPE-DMAB对OONO-具有高选择性。
以二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)为包封基质,将TPE-DMAB制备成纳米粒子。图12中a示出了TPE-DMAB纳米粒子对ONOO-的荧光响应。如图12中b所示,通过动态光散射(DLS)发现纳米粒子的流体动力学直径约为150纳米。透射电子显微镜(TEM)观察表明,纳米粒子呈现均匀的球形,其大小与DLS结果统一。
随后,发明人测定了TPE-DMAB纳米粒子的紫外吸收和其对OONO-和的荧光响应。TPE-DMAB纳米粒子显示出与TPE-DMAB分子相似的紫外吸收(图13)。在pH7.4时进行检测,TPE-DMAB纳米粒子对OONO-的选择性也很高(图14)。TPE-DMAB纳米粒子也可以在6.8到7.4的pH范围内检测OONO-(图15)。如图12中c所示,荧光强度随着OONO-含量的增加而逐渐增强。值得注意的是,OONO-在炎症区域的产生速率可达50-100微摩尔/分钟。因此,TPE-DMAB纳米粒子也非常适合用于炎症区域的可视化。
随后TPE-DMAB纳米粒子被用于检测细胞内源性产生的OONO-。经细菌细胞壁脂多糖(LPS)和巴豆醇-12-十四烷酸酯-13-乙酸酯(PMA)刺激后的小鼠巨噬细胞系RAW264.7可诱导产生OONO-。与TPE-DMAB纳米粒子在37℃孵育3小时后,从原始RAW264.7细胞中几乎不容易观察到荧光(图16中a)。而LPS/PMA处理的细胞表现出强烈的荧光(图16中b),表明在LPS/PMA处理的RAW264.7细胞会实现探针的开启检测。进一步证实荧光增强是由内源性生成的OONO-引起的。乙酰半胱氨酸(NAC)是一种天然的RONS清除剂,在LPS/PMA处理的巨噬细胞中。与NAC预孵育2h后,巨噬细胞仅可见微弱荧光(图16中c)。这些结果表明内源性产生的OONO-能激发生物探针的荧光,说明TPE-DMAB纳米粒子适用于应激细胞OONO-的检测。此外,通过细胞存活率分析(MTT)实验(图17)评估所呈现的探针的细胞毒性结果表明生物探针具有良好的生物相容性。
在验证TPE-DMAB纳米粒子对应激巨噬细胞内源性OONO-的检测能力后,将其应用于体内炎症部位的特异性成像。在炎症过程中,免疫细胞如中性粒细胞和巨噬细胞会聚集到炎症部位并被刺激释放大量的OONO-。为了验证TPE-DMAB纳米粒子能否在体内用于OONO-成像,选择LPS诱导的炎症小鼠并用TPE-DMAB纳米粒子孵育。如图18中a所示,在注射生物探针之前可以检测到微弱的荧光信号,而在注射TPE-DMAB纳米粒子30分钟后捕获到强烈的荧光信号(图18中b)。此外,为了证实TPE-DMAB纳米粒子的RONS特异性活化,在注射纳米粒子前3小时注射了天然RONS清除剂尿酸,如图18中c所示,注射RONS清除剂的小鼠几乎没有可以检测到的荧光信号。图19示出了注射TPE-DMAB纳米粒子的小鼠感染皮肤中的定量平均荧光。这些结果证明,TPE-DMAB纳米粒子是非常理想的炎症部位的荧光生物探针。
由此描述了本发明主题,显然可以以许多方式对本发明主题进行修改或改变。这些修改和改变不应被认为是脱离本发明主题的精神和范围,并且旨在将所有这些修改和改变包括在所附权利要求的范围内。
Claims (10)
3.一种纳米颗粒,所述纳米颗粒包括根据权利要求1或2所述的化合物。
4.根据权利要求3所述的纳米颗粒,所述纳米颗粒以二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)为包封基质。
5.根据权利要求1或2所述的化合物或根据权利要求3或4所述的纳米颗粒用于检测过氧亚硝酸阴离子的应用。
6.根据权利要求5所述的应用,其特征在于,所述化合物对其他活性氮氧类物质无反应,所述其他活性氮氧类物质优选包括ClO-、H2O2、·OH、tBuOOH、·OBu、O2 -、1O2、NO2 -和NO3 -。
7.根据权利要求5或6所述的应用,其特征在于,所述检测过氧亚硝酸阴离子包括检测所述化合物与过氧亚硝酸阴离子反应后的产物发射的荧光。
8.根据权利要求5或6所述的应用,其特征在于,所述检测过氧亚硝酸阴离子包括在细胞水平检测活性氮氧类物质。
9.根据权利要求5-8任一项所述的应用,其特征在于,所述检测过氧亚硝酸阴离子包括在pH大于6.8(优选pH6.8-pH7.4)下检测过氧亚硝酸阴离子。
10.权利要求5-9任一项所述的应用,其特征在于,所述应用包括对炎症相关的吞噬细胞进行染色,或对炎症部位进行显示。
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