CN116217515A - 一种化合物及其制备方法和在检测半胱氨酸中的应用 - Google Patents
一种化合物及其制备方法和在检测半胱氨酸中的应用 Download PDFInfo
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Abstract
Description
技术领域
本发明涉及一种化合物及其制备方法和在检测半胱氨酸中的应用。
背景技术
半胱氨酸是一种含硫的蛋白质源性氨基酸,是细胞结构和信号传输的关键组成部分,是从气体硫化氢分子到辅酶A等各种生物分子的必备前体。尽管是细胞中最不丰富的氨基酸,但其却承担着最大数量的蛋白质翻译后的修饰任务,而且作为抗氧化剂谷胱甘肽和其他潜在抗氧化剂的组成部分,它在氧化还原平衡体系中发挥着举足轻重的作用。一旦半胱氨酸在细胞内的水平出现异常,神经毒性、毛发脱色、水肿、肝损伤、皮肤病变、恶性肿瘤等多种疾病可能会接踵而至。
研究表明,细胞已经进化出多种机制来维持动态平衡的半胱氨酸供应以便满足多种生命活动的需求。虽然半胱氨酸有多种获取途径,但它也会被各种代谢途径消耗,例如蛋白质合成或生成体内的含硫分子,如谷胱甘肽、牛磺酸、硫化双丙酸、辅酶A和重要的气体信号分子硫化氢。
现有的研究数据还表明,半胱氨酸的稳态与无数生理过程息息相关,半胱氨酸在神经毒性、阿尔茨海默病、和心血管疾病等许多疾病中过度表达,特别是肿瘤部位,它甚至可以反映不同阶段的肿瘤组织。所以,实时无创检测体内半胱氨酸水平的变化,对于了解肿瘤的病理功能及监测癌症治疗至关重要。
目前,用于半胱氨酸检测的探针大多基于单一的荧光成像,但由于光散射太强、组织穿透深度不够等弊端限制了它在深层组织中半胱氨酸的检测应用。相比之下,光声探针结合了激光的选择性和声波的深穿透效应,从而实现了对组织的高空间分别率的成像。高对比度光声传感成像(High-Contrast Photoacoustic Sensing Imaging,PASI)在很大程度上取决于吸收剂的光学吸收变化,通常由可激活探针通过可控地将吸收的电磁能转换为超声波来实现。然而,由于它们在近红外(Near Infrared Region,NIR)区域的吸收光谱变化很小,目前大多数光声探针对特定物种的成像对比度仍然受限。
发明内容
本发明的目的在于提供一种高稳定性的化合物,其结构中的丙烯酸酯部分通过一个氨基甲酸酯键彻底淬灭染料亚甲基蓝的光声和荧光效应,半胱氨酸响应后,彻底恢复亚甲基蓝的光声和荧光特征,从而可以用于体外、细胞和活体内半胱氨酸的荧光和光声成像分析,具有灵敏度高的特点。
本发明的目的通过下述技术方案实现:
一种化合物,其结构如式I所示:
所述化合物的制备方法,包括如下步骤:
(1)将亚甲基蓝、碳酸氢钠和连二亚硫酸钠加入到混合溶剂中,在40~60℃下进行反应,待反应结束后冷却至室温,分液,取有机溶剂层,得到溶液A;
(2)在冰浴条件下,将三乙胺加入到步骤(1)中得到的溶液A中,再滴加PB-Cl(4-丙烯胺酸酯苄基酰氯,4-(((chlorocarbonyl)oxy)methyl)phenyl acrylate)溶液进行反应,待反应结束后萃取,洗涤、干燥,减压浓缩,得到式I所示的化合物;
上述过程发生以下反应:
步骤(1)中所述的亚甲基蓝、碳酸氢钠和连二亚硫酸钠的摩尔比为1:(3~4):(2~4);优选为1:3.48:2.85。
步骤(1)中所述的混合溶剂为有机溶剂与水混合得到的溶剂;优选为有机溶剂和水按体积比为4:1混合得到的溶剂。
所述的有机溶剂包括甲苯、苯等;优选为甲苯。
步骤(1)中所述的混合溶剂的用量优选为按每毫升(ml)混合溶剂配比7.5mg亚甲基蓝计算。
步骤(1)中所述的反应的时间优选为2~4h。
步骤(2)中所述的三乙胺、PB-Cl与所述亚甲基蓝的摩尔比为3.5:1.5:1。
步骤(2)中所述的PB-Cl通过如下方法制备得到:将4-(羟甲基)苯基丙烯酸酯、三光气和三乙胺溶于四氢呋喃中,冰浴条件下搅拌反应,待反应结束后萃取、干燥、浓缩,得到PB-Cl。
所述的4-(羟甲基)苯基丙烯酸酯、三光气和三乙胺的摩尔比为(1.2~2):1:(2~4);摩尔比优选为1.6:1:3.68。
所述的四氢呋喃的用量优选为按每毫升(ml)四氢呋喃配比35.6mg 4-(羟甲基)苯基丙烯酸酯计算。
所述的搅拌反应的时间为1~3h;优选为1小时。
所述的萃取为采用二氯甲烷与水混合得到的溶剂进行萃取;优选为采用二氯甲烷与水按比为1:1混合得到的溶剂进行萃取。
所述的干燥为采用无水硫酸钠进行干燥。
步骤(2)中所述的萃取为采用二氯甲烷与水混合得到的溶剂进行萃取;优选为采用二氯甲烷与水按比为1:1混合得到的溶剂进行萃取。
步骤(2)中所述的洗涤为用水洗涤;优选为用水洗3次以上。
步骤(2)中所述的干燥为采用无水硫酸镁进行干燥。
所述化合物的制备方法,还包括将步骤(2)中获得的化合物进行分离纯化的步骤。
所述的分离纯化为采用硅胶色谱柱进行分离纯化。
所述的化合物可以应用在体外、活细胞及活体原位半胱氨酸的成像检测中,基于半胱氨酸对丙烯酸酯基团的特异性识别,使丙烯酸酯基团水解为酚羟基,淬灭基团丙烯酸的离去使亚甲基蓝恢复,在近红外光的照射下,产生较强的光声/荧光信号,实现半胱氨酸的高灵敏光声/荧光检测,并实现对细胞及原位肿瘤内半胱氨酸的高对比度光声/荧光双模态成像分析。
所述的化合物可以应用在体外、细胞及活体肿瘤内半胱氨酸的光声/荧光双模态成像分析中。
所述的细胞包括在EMT6(鼠乳腺癌细胞)、PC12(源自大鼠肾上腺髓质的嗜铬细胞瘤)和U87(人神经胶质瘤细胞)等。
所述的活体肿瘤为EMT6小鼠肿瘤模型。
所述的化合物的浓度优选为10μM。
本发明相对于现有技术具有如下的优点及效果:
1、本发明基于亚甲基蓝的式I化合物的合成是通过将能够被半胱氨酸识别的丙烯酸酯部分缀合到临床使用的亚甲基蓝上构建而成,合成简单,易于后处理;同时,由于结构中丙烯酸酯部分与亚甲基蓝的非共轭结构形式存在分子内极性平衡,产物稳定性高,这是本专利所涉及式I化合物的首要特色之一。
2、本发明所涉及的分子探针(式I化合物)其响应原理是丙烯酸酯部分通过一个氨基甲酸酯键彻底淬灭染料亚甲基蓝的光声和荧光效应,半胱氨酸响应后,得到临床可用的亚甲基蓝,彻底恢复亚甲基蓝的光声和荧光特征,用于体外、细胞和活体内半胱氨酸的荧光和光声成像分析具有灵敏度高的特点。
3、本发明获得的式I化合物分子探针选择性好,可特异性响应半胱氨酸后得到临床可用的有机染料亚甲基蓝。
4、本发明基于亚甲基蓝的式I化合物分子探针克服了传统单一荧光成像的半胱氨酸检测灵敏度低以及在活体检测中成像深度浅、对比度低的问题。
5、本发明所涉及的式I化合物分子探针在近红外区域几乎没有光学吸收和荧光发射,但当该探针通过与丙烯酸酯的环化反应被半胱氨酸激活后,可实现近红外吸收和荧光由“关到开”转变,因此,可介导对半胱氨酸的高对比度光声/荧光成像分析,且由于亚甲基蓝具有光敏特性,可用于半胱氨酸高表达病灶组织的靶向光动力治疗。
附图说明
图1为本发明实施例1中式I化合物的液相色谱图。
图2为本发明实施例1中式I化合物的电感耦合等离子体质谱图。
图3为本发明实施例2中式I化合物与半胱氨酸反应后产物的电感耦合等离子体质谱图;其中,图A为反应后产物亚甲基蓝的质谱图,图B为反应后副产物的质谱图。
图4为本发明实施例3中式I化合物与不同浓度半胱氨酸反应前后的吸收光谱图。
图5为本发明实施例4中式I化合物与不同浓度半胱氨酸反应前后的荧光光谱图。
图6为本发明实施例5中式I化合物与不同浓度半胱氨酸(a)以及不同反应干扰物种(b)前后的光声成像图和光声信号强度图。
图7为本发明实施例6中式I化合物在不同细胞中不同浓度下的细胞活力图。
图8为本发明实施例7中式I化合物在光照EMT6细胞中不同浓度下的细胞活力图。
图9为本发明实施例8中式I化合物对EMT6细胞内源性半胱氨酸的光声检测结果图。
图10为本发明实施例9中式I化合物对EMT6细胞内源性半胱氨酸的荧光成像检测结果图。
图11为本发明实施例10中式I化合物的体内光声成像图以及光声信号强度图;其中,图A为光声成像图;图B为光声信号强度图。
图12为本发明实施例11中式I化合物的肿瘤靶向的体内荧光成像图和荧光信号强度图。
图13为本发明实施例11中式I化合物的生物分布荧光图及数据统计结果。
具体实施方式
下面结合实施例及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1:式I化合物的制备
式I化合物的制备方法,具体包括如下步骤:
将亚甲基蓝(373.9mg,1mmol),碳酸氢钠(292.4mg,3.48mmol)和连二硫酸钠(496.2mg,2.85mmol)加入到40ml甲苯和10ml水形成的混合溶液中,60℃反应2小时,冷至室温,分液,保留甲苯层;然后在冰浴条件下,往甲苯溶液中,加入三乙胺(354.2mg,3.5mmol),并将溶解在40ml甲苯中的PB-Cl(360mg,1.5mmol)的溶液缓慢滴加到上述溶液中。滴加完毕,室温过夜反应,减压浓缩,加入水(100ml)和二氯甲烷(100ml)萃取,有机相再用水洗3次。收集有机相,加入无水硫酸镁干燥,浓缩,用硅胶色谱柱进行分离纯化得到式I化合物。其中PB-Cl的制备如下:
将4-(羟甲基)苯基丙烯酸酯(768mg,3.2mmol)和三光气(593.5mg,2mmol)溶于20ml四氢呋喃中,在冰浴条件下加入三乙胺(744.8mg,7.36mmol),冰浴搅拌反应1h后,用二氯甲烷和水萃取(体积比为1:1),用无水硫酸钠干燥,浓缩有机相,得到PB-Cl(4-丙烯胺酸酯苄基酰氯,4-(((chlorocarbonyl)oxy)methyl)phenyl acrylate)。
式I化合物表征数据:
液相色谱:3.6min(甲醇/乙腈)(图1)
质谱图(图2):calcd.for[M+]490,found:490.
实施例2:式I化合物与半胱氨酸反应后产物的电感耦合等离子体质谱图
配制实施例1中获得的式I化合物以及半胱氨酸水溶液,然后将式I化合物与半胱氨酸水溶液混合反应二十分钟后测定其电感耦合等离子体质谱;其中,反应体系中式I化合物的浓度为10μM,半胱氨酸的浓度为100μM。质谱如图3所示,从图中可以看出式I化合物在与半胱氨酸反应后会产生亚甲基蓝和环化副产物。
实施例3:式I化合物与不同浓度半胱氨酸反应前后的吸收光谱
配制实施例1中获得的式I化合物溶液以及半胱氨酸水溶液,然后将式I化合物溶液与半胱氨酸水溶液混合反应三十分钟后测定其吸收;其中,反应体系中式I化合物的浓度为10μM,半胱氨酸的浓度为0~100μM。吸收光谱如图4所示,从图中可以看出式I化合物随着半胱氨酸浓度增加,吸收系数也逐渐增强。
实施例4:式I化合物与不同浓度半胱氨酸反应前后的荧光光谱
配制实施例1中获得的式I化合物溶液以及半胱氨酸水溶液,然后将式I化合物溶液与半胱氨酸水溶液混合反应三十分钟后测定其荧光;其中,反应体系中式I化合物的浓度为10μM,半胱氨酸的浓度为0~100μM。荧光光谱如图5所示,从图中可以看出式I化合物随着半胱氨酸浓度增加,荧光强度也逐渐增强。表明探针可以作为响应半胱氨酸的荧光探针。
实施例5:式I化合物与不同浓度半胱氨酸反应后光声成像和光声信号强度图
配制浓度为10μM式I化合物溶液共6组,以及浓度为0,20,40,60,80,100μM的半胱氨酸水溶液,然后将式I化合物溶液与半胱氨酸水溶液混合后进行反应(式I化合物溶液的用量为40μl,半胱氨酸水溶液的用量分别为1,2,3,4,5μl),每组各自反应20分钟后,用光声计算机断层扫描仪测定6组溶液的光声信号,以及光声二维图(图6A)。从图中可以看出光声信号随浓度的增加,665nm的光声信号强度逐渐增强。表明式I化合物可以作为响应半胱氨酸的光声探针。
另配制浓度为10μM式I化合物溶液共8组,然后将式I化合物溶液分别与100μM半胱氨酸或抗坏血酸(AA)、硫氢化钠(NaHS)、丙氨酸(Ala)、甘氨酸(Gly)、丝氨酸(Ser)、谷胱甘肽(GSH)、高半胱氨酸(Hcy)混合后进行反应(式I化合物溶液的用量为40μl,半胱氨酸及干扰物溶液用量分别为5μl),每组各自反应20分钟后,用光声计算机断层扫描仪测定6组溶液的光声信号,以及光声二维图(图6B),结果表明式I化合物可选择性响应半胱氨酸。
实施例6:式I化合物的细胞毒性分析
利用MTT试剂测定细胞的相对活力。分别将EMT6细胞和PC12细胞以2×103/孔的密度接种到96孔板中,恒温培养24h。然后设置6组实验,每组实验设置5组平行实验。(1)对照组:不用式I化合物处理;(2)探针组:用不同浓度的式I化合物溶液处理(10μΜ,20μΜ,30μΜ,40μΜ,50μΜ)。每组细胞在黑暗条件下于恒温培养箱中孵育24h,分别用PBS洗涤3次,加入新的培养液和10%的MTT(5mg/mL)溶液,黑暗条件下于恒温培养箱中孵育4h,再加入100μL DMSO/孔,黑暗条件下于恒温培养箱中孵育4h,用酶标仪测定每孔细胞在490nm处的吸光度值。细胞平均相对活力值按以下公式计算:细胞平均相对活力(%)=处理组细胞平均吸光度值/对照组细胞平均吸光度值×100%。结果表明如图7所示式I化合物具有较好的生物相容性。
实施例7:式I化合物的光敏活性分析
将癌细胞ETM6孵育在六孔板内,配制不同浓度的式I化合物溶液(0,2,4,6,8,10)加入上述细胞培养液中,其中0为空白对照组,继续孵育1h,再将细胞经633激光照射后进行MTT细胞活力测试。如图8所示,结果表明式I化合物I可以在癌细胞内半胱氨酸的作用下,释放光敏剂亚甲基蓝,进行光动力治疗。
实施例8:式I化合物在癌细胞EMT6内光声响应的统计情况
将癌细胞EMT6孵育在不同培养皿,配制浓度为10μM式I化合物溶液,分为8组,加入上述细胞培养液中,其中0为空白对照组,继续孵育1h;再在上述溶液中加入不同浓度的半胱氨酸溶液(0,50μM,100μM,150μM,250μM,300μM,350μM)其中,式I化合物溶液和半胱氨酸的用量均为40μl。将细胞洗涤消化后,用光声计算机断层扫描仪测定8组溶液的光声信号,以及光声二维图。结果如图9所示,从图中可以看出式I化合物可以对细胞内半胱氨酸有很好光声响应。
实施例9:式I化合物在癌细胞HepG2内荧光响应的统计情况
将癌细胞HepG2孵育在共聚焦皿内,配制浓度为10μM式I化合物溶液,分为5组,第一组不作处理(作为对照),第二组加入式I化合物溶液(probe),第三组至第五组分别加入式I化合物溶液以及不同浓度的半胱氨酸(50μM,100μM,150μM,);其中,式I所示的化合物溶液和半胱氨酸的用量均为40μl。继续孵育1h,用荧光共聚焦显微镜进行细胞荧光成像。结果如图10所示,从图中可以看出式I化合物可以对细胞内半胱氨酸有很好的荧光响应。
实施例10:式I化合物在活体内的光声成像。
将小鼠(3~4周BALB/c鼠,平均2体重20g,购买于南方医科大学)分为三组,第一组:提前6小时在瘤内注射NEM(1mM,30μL),后通过尾静脉注射100μL式I化合物溶液(10μM);第二组:提前6小时在瘤内注射半胱氨酸(Cys,150μM,30μL),后通过尾静脉注射100μL式I化合物溶液;第三组:直接通过尾静脉注射100μL式I化合物溶液;然后分别在注射前注射后0、3、6、9、12和24小时的时间点用光声断层扫描成像***进行光声成像,最后将光声成像所得数据与超声图片进行叠加后进行数据的统计分析。结果如图11所示,结果表明式I化合物在小鼠体内代谢到肿瘤部位后可对体内内源性半胱氨酸进行选择性光声成像。
实施例11:式I化合物在活体内的荧光成像
将小鼠(3~4周BALB/c裸鼠,平均2体重20g,购买于南方医科大学)先用5%水合氯醛麻醉。将式I化合物(100μL,4.5mg/kg)静脉注射到小鼠中,然后分别在注射后0、3、6、9、12和24小时的时间点用近红外成像***(Odyssey LI-COR,USA)进行荧光成像和数据统计分析。(激发波长为680nm,且在720nm的波长下采集信号)。结果如图12所示,结果表明式I化合物在小鼠体内代谢到肿瘤部位后可对体内内源性半胱氨酸进行选择性荧光成像。
另外,在小鼠被注射探针12小时后,对其主要器官(心、肝、脾、肺、肾)进行了收集和成像。鉴于肿瘤细胞内高表达的半胱氨酸,结果如图13显示,式I化合物具有很好的肿瘤靶向性。
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)
2.权1所述化合物的制备方法,其特征在于包括如下步骤:
(1)将亚甲基蓝、碳酸氢钠和连二亚硫酸钠加入到混合溶剂中,在40~60℃下进行反应,待反应结束后冷却至室温,分液,取有机溶剂层,得到溶液A;
(2)在冰浴条件下,将三乙胺加入到步骤(1)中得到的溶液A中,再滴加4-丙烯胺酸酯苄基酰氯溶液进行反应,待反应结束后萃取,洗涤、干燥,减压浓缩,得到式I所示的化合物。
3.根据权利要求2所述的制备方法,其特征在于:步骤(1)中所述的亚甲基蓝、碳酸氢钠和连二亚硫酸钠的摩尔比为1:(3~4):(2~4)。
4.根据权利要求2所述的制备方法,其特征在于:步骤(1)中所述的混合溶剂为有机溶剂与水混合得到的溶剂。
5.根据权利要求2所述的制备方法,其特征在于:步骤(2)中所述的三乙胺、PB-Cl与所述亚甲基蓝的摩尔比为3.5:1.5:1。
6.根据权利要求2所述的制备方法,其特征在于:步骤(2)中所述的4-丙烯胺酸酯苄基酰氯通过如下方法制备得到:将4-(羟甲基)苯基丙烯酸酯、三光气和三乙胺溶于四氢呋喃中,冰浴条件下搅拌反应,待反应结束后萃取、干燥、浓缩,得到4-丙烯胺酸酯苄基酰氯。
7.根据权利要求6所述的制备方法,其特征在于:所述的4-(羟甲基)苯基丙烯酸酯、三光气和三乙胺的摩尔比为(1.2~2):1:(2~4)。
8.根据权利要求2所述的制备方法,其特征在于:所述化合物的制备方法,还包括将步骤(2)中获得的化合物进行分离纯化的步骤。
9.权1所述的化合物在体外、活细胞及活体原位半胱氨酸的成像检测中的应用。
10.权1所述的化合物在体外、细胞及活体肿瘤内半胱氨酸的光声/荧光双模态成像分析中的应用。
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Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050214807A1 (en) * | 2003-11-19 | 2005-09-29 | Iain Johnson | Environmental sensitive fluorogenic compounds and their application for singlet oxygen and protein detection |
US20080295259A1 (en) * | 2007-02-22 | 2008-12-04 | Asahi Kasei Pharma Corporation | Method for stabilizing leuco-type colorant |
CN105431533A (zh) * | 2013-07-09 | 2016-03-23 | 协和梅迪克斯株式会社 | 糖化六肽氧化酶及其用途 |
CN108456197A (zh) * | 2018-03-23 | 2018-08-28 | 华南师范大学 | 用于活体检测硫化氢的光声比率纳米探针及其制备方法与应用 |
CN108884386A (zh) * | 2016-01-26 | 2018-11-23 | 特拉维夫大学拉玛特有限公司 | 用于诊断和体内成像的化学发光探针 |
CN109503515A (zh) * | 2018-11-16 | 2019-03-22 | 山西大学 | 一种亚甲基蓝衍生物及其合成方法和应用 |
CN110050069A (zh) * | 2016-10-12 | 2019-07-23 | 爱-森斯株式会社 | 血红蛋白a1c定量分析试剂盒 |
CN113354584A (zh) * | 2021-06-15 | 2021-09-07 | 郑州大学 | 一种区分Cys、Hcy和GSH的萘酰亚胺类荧光探针及其制备方法与应用 |
CN114736199A (zh) * | 2022-05-25 | 2022-07-12 | 山西大学 | 一种基于亚甲基蓝的近红外荧光探针及其合成方法和应用 |
CN114835698A (zh) * | 2022-05-23 | 2022-08-02 | 南京林业大学 | 一种用于检测半胱氨酸的(二苯氨基)苯基黄酮类荧光探针及其制备方法 |
CN114835658A (zh) * | 2022-04-02 | 2022-08-02 | 华南师范大学 | 一种用于检测硫化氢的荧光探针及其制备方法和应用 |
KR20220165891A (ko) * | 2021-06-08 | 2022-12-16 | 제이투에이치바이오텍 (주) | 3성분 프로드럭, 이의 약학적 조성물 및 의약 용도 |
-
2022
- 2022-12-20 CN CN202211643235.3A patent/CN116217515B/zh active Active
Patent Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050214807A1 (en) * | 2003-11-19 | 2005-09-29 | Iain Johnson | Environmental sensitive fluorogenic compounds and their application for singlet oxygen and protein detection |
US20080295259A1 (en) * | 2007-02-22 | 2008-12-04 | Asahi Kasei Pharma Corporation | Method for stabilizing leuco-type colorant |
CN105431533A (zh) * | 2013-07-09 | 2016-03-23 | 协和梅迪克斯株式会社 | 糖化六肽氧化酶及其用途 |
CN108884386A (zh) * | 2016-01-26 | 2018-11-23 | 特拉维夫大学拉玛特有限公司 | 用于诊断和体内成像的化学发光探针 |
CN110050069A (zh) * | 2016-10-12 | 2019-07-23 | 爱-森斯株式会社 | 血红蛋白a1c定量分析试剂盒 |
CN108456197A (zh) * | 2018-03-23 | 2018-08-28 | 华南师范大学 | 用于活体检测硫化氢的光声比率纳米探针及其制备方法与应用 |
CN109503515A (zh) * | 2018-11-16 | 2019-03-22 | 山西大学 | 一种亚甲基蓝衍生物及其合成方法和应用 |
KR20220165891A (ko) * | 2021-06-08 | 2022-12-16 | 제이투에이치바이오텍 (주) | 3성분 프로드럭, 이의 약학적 조성물 및 의약 용도 |
CN113354584A (zh) * | 2021-06-15 | 2021-09-07 | 郑州大学 | 一种区分Cys、Hcy和GSH的萘酰亚胺类荧光探针及其制备方法与应用 |
CN114835658A (zh) * | 2022-04-02 | 2022-08-02 | 华南师范大学 | 一种用于检测硫化氢的荧光探针及其制备方法和应用 |
CN114835698A (zh) * | 2022-05-23 | 2022-08-02 | 南京林业大学 | 一种用于检测半胱氨酸的(二苯氨基)苯基黄酮类荧光探针及其制备方法 |
CN114736199A (zh) * | 2022-05-25 | 2022-07-12 | 山西大学 | 一种基于亚甲基蓝的近红外荧光探针及其合成方法和应用 |
Non-Patent Citations (7)
Title |
---|
WEI, WENYU,等: "HDAC6-Activatable Multifunctional Near-Infrared Probe for Glioma Cell Detection and Elimination", ANALYTICAL CHEMISTRY, vol. 96, no. 06, 16 February 2024 (2024-02-16), pages 2406 - 2414 * |
XIE, S,等: "High-Contrast Photoacoustic Imaging of Localized Cysteine in Orthotopic Breast Cancer Enabled by A Totally-Caged Methylene Blue Probe", CHEMISTRY-A EUROPEAN JOURNAL, vol. 30, no. 11, 21 February 2024 (2024-02-21), pages 1 - 9 * |
姚书帆;尧雨斯;郑武斌;叶晨喆;应杰;吕光磊;李春霞;: "基于亚甲基蓝的近红外荧光探针用于HOCl的特异性检测", 发光学报, no. 07, 15 July 2020 (2020-07-15), pages 42 - 50 * |
孙志斌;郜梦娇;孟雅莉;尚亚靖;亢延飞;: "高选择性半胱氨酸荧光探针的合成及其在活细胞中的应用", 河北北方学院学报(自然科学版), no. 01, 28 January 2020 (2020-01-28), pages 23 - 31 * |
廖旭芳: "基于亚甲基蓝,萘酰亚胺荧光团检测HClO、H2O2、Cys的荧光探针设计合成及细胞成像研究", 中国优秀硕士学位论文全文数据库 电子期刊, no. 01, 15 January 2023 (2023-01-15), pages 006 - 101 * |
程威: "有机小分子荧光探针的设计及其在活性硫氧组分检测中的应用", 中国优秀硕士学位论文全文数据库 电子期刊, no. 01, 31 January 2021 (2021-01-31), pages 014 - 1482 * |
蒋凯;曹梁;郝志峰;陈美燕;程洁銮;李晓;肖萍;陈亮;汪朝阳;: "苯并噻唑基荧光探针的设计、合成与应用研究进展", 有机化学, no. 09, 30 September 2017 (2017-09-30), pages 60 - 75 * |
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