反应激活型疾病诊断荧光探针及其它相关探针的设计、合成及应用
发布时间：2018-12-13 01:45
【摘要】：本文设计、合成了三类基于反应激活的荧光探针,反应前后荧光团的电子云密度得到重新排列,使得反应前后可以产生两个不同波段的荧光发射峰,因此可以对检测对象实现比例检测,大大提高了检测的精度和可信度。 1.设计、合成了N-乙酰转移酶II特异性荧光探针Amonafide。临床代谢研究发现,化合物amonafide在体内主要被N-乙酰基转移酶II催化代谢生成Acetyl-Amonafide。基于此,设计了化合物Amonafide作为N-乙酰转移酶II特异性荧光探针。化合物Amonafide和Acetyl-Amonafide在350 nm有共同的最大吸收峰,而发射波长分别在460 nm和580 nm,更重要的是代谢产物Acetyl-Amonafide的荧光量子产率高达0.74。酶水平和细胞水平实验进一步证明了探针Amonafide对NAT2的高选择性和灵敏性(1 nM)。并且成功地将探针Amonafide应用于HepG2细胞株内的N-乙酰转移酶II荧光成像检测,这为芳胺类药物的个性化给药奠定了基础。同时完成了分别针对N-乙酰转移酶I和N-乙酰转移酶II的长波长荧光探针的设计和部分合成工作。 2.以4-氨基-1,8-萘酰亚胺为荧光团母体,通过异氰酸酯键与受体对硝基苄醇链接,设计了针对肿瘤乏氧的比率型荧光探针RHP。通过ICT原理,可以使反应后的最大荧光发射波长红移(475nm到550 nm)。详细研究了探针RHP对硝基还原酶(NTR)的荧光响应和动力学曲线,同时检测了内源性还原物质对探针的影响。实验结果进一步证明了探针RHP对硝基还原酶的高选择性。乏氧／有氧实验证明,当GrHy/GrAr和B1Hy/B1Ar的比值在45以上时,细胞即处于乏氧环境。探针RHP可以作为肿瘤乏氧诊断探针用于体内实体瘤的早期诊断。同时通过乏氧原理设计并合成了基于Amonafide的前药化合物。 3.以苯并噻唑类化合物(HBTBC)为荧光团母体,设计了一类基于ESIPT的荧光探针。反应前后两种化合物的斯托克斯位移高达120 nm以上,且光谱交叉的区域少,设计的探针检测结果的可信度和准确度更高。以Tsuji-Trost烯丙基的氧化反应为基础设计了针对Pd(0)的荧光探针OHBT。当OHBT与Pd(0)反应后,最大发射波长由410 nm红移到550 nm,设计的探针对Pd(0)有着高度的选择性。同时通过滤纸对Pd(0)实现了固体荧光检测,并成功对Suzuki偶联反应产物中残留的Pd(0)进行了定量检测。
[Abstract]:Three kinds of fluorescence probes based on reactive activation were designed and synthesized in this paper. The electron cloud density of the fluorescence clusters was rearranged before and after the reaction so that two fluorescence emission peaks in different bands could be generated before and after the reaction. Therefore, the scale detection can be realized, and the accuracy and credibility of detection can be greatly improved. 1. N-acetyltransferase II specific fluorescent probe Amonafide. was designed and synthesized. Clinical metabolic studies show that the compound amonafide is mainly metabolized by N-acetyltransferase II to produce Acetyl-Amonafide. in vivo. Based on this, the compound Amonafide was designed as a specific fluorescence probe for N-acetyltransferase (N-acetyltransferase) II. The compound Amonafide and Acetyl-Amonafide have the same maximum absorption peak at 350 nm, and the emission wavelengths are 460 nm and 580 nm, respectively. More importantly, the fluorescence quantum yield of the metabolite Acetyl-Amonafide is up to 0.74. Enzyme level and cell level experiments further demonstrated the high selectivity and sensitivity of probe Amonafide to NAT2 (1 nM). The probe Amonafide was successfully applied to the detection of N-acetyltransferase II in HepG2 cell line by fluorescence imaging, which laid a foundation for individualized administration of aromatic amines. At the same time, the design and partial synthesis of long wavelength fluorescence probe for N- acetyltransferase I and N- acetyltransferase II were completed. 2. The ratio fluorescence probe RHP. for tumor hypoxia was designed by linking isocyanate bond with the receptor p-nitrobenzyl alcohol using 4-amino-1-butadiene-8-naphthalimide as the fluorescence group parent. The maximum fluorescence emission wavelength (475nm to 550 nm).) can be shifted by the principle of ICT. The fluorescence response and kinetic curves of probe RHP to nitroreductase (NTR) were studied in detail. The results further demonstrated the high selectivity of probe RHP for nitroreductase. Hypoxia / aerobic experiments showed that when the ratio of GrHy/GrAr to B1Hy/B1Ar was above 45, the cells were in hypoxic environment. The probe RHP can be used as a diagnostic probe for tumor hypoxia in early diagnosis of solid tumors in vivo. At the same time, prodrug compounds based on Amonafide were designed and synthesized by hypoxia principle. 3. A kind of fluorescence probe based on ESIPT was designed by using benzothiazole compound (HBTBC) as the fluorescence group parent. The Stokes shift of the two compounds before and after the reaction is more than 120 nm, and the cross region of the spectra is less, so the detection results of the designed probe are more reliable and accurate. Based on the oxidation of Tsuji-Trost allyl, a fluorescent probe OHBT. for Pd (0) was designed. When OHBT reacts with Pd (0), the maximum emission wavelength shifts from 410 nm red to 550 nm, with high selectivity to Pd (0). At the same time, Pd (0) was detected by filter paper, and the residual Pd (0) in the coupling product of Suzuki was detected quantitatively.
【学位授予单位】：华东理工大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R346;TP212.2
本文编号：2375645
[Abstract]:Three kinds of fluorescence probes based on reactive activation were designed and synthesized in this paper. The electron cloud density of the fluorescence clusters was rearranged before and after the reaction so that two fluorescence emission peaks in different bands could be generated before and after the reaction. Therefore, the scale detection can be realized, and the accuracy and credibility of detection can be greatly improved. 1. N-acetyltransferase II specific fluorescent probe Amonafide. was designed and synthesized. Clinical metabolic studies show that the compound amonafide is mainly metabolized by N-acetyltransferase II to produce Acetyl-Amonafide. in vivo. Based on this, the compound Amonafide was designed as a specific fluorescence probe for N-acetyltransferase (N-acetyltransferase) II. The compound Amonafide and Acetyl-Amonafide have the same maximum absorption peak at 350 nm, and the emission wavelengths are 460 nm and 580 nm, respectively. More importantly, the fluorescence quantum yield of the metabolite Acetyl-Amonafide is up to 0.74. Enzyme level and cell level experiments further demonstrated the high selectivity and sensitivity of probe Amonafide to NAT2 (1 nM). The probe Amonafide was successfully applied to the detection of N-acetyltransferase II in HepG2 cell line by fluorescence imaging, which laid a foundation for individualized administration of aromatic amines. At the same time, the design and partial synthesis of long wavelength fluorescence probe for N- acetyltransferase I and N- acetyltransferase II were completed. 2. The ratio fluorescence probe RHP. for tumor hypoxia was designed by linking isocyanate bond with the receptor p-nitrobenzyl alcohol using 4-amino-1-butadiene-8-naphthalimide as the fluorescence group parent. The maximum fluorescence emission wavelength (475nm to 550 nm).) can be shifted by the principle of ICT. The fluorescence response and kinetic curves of probe RHP to nitroreductase (NTR) were studied in detail. The results further demonstrated the high selectivity of probe RHP for nitroreductase. Hypoxia / aerobic experiments showed that when the ratio of GrHy/GrAr to B1Hy/B1Ar was above 45, the cells were in hypoxic environment. The probe RHP can be used as a diagnostic probe for tumor hypoxia in early diagnosis of solid tumors in vivo. At the same time, prodrug compounds based on Amonafide were designed and synthesized by hypoxia principle. 3. A kind of fluorescence probe based on ESIPT was designed by using benzothiazole compound (HBTBC) as the fluorescence group parent. The Stokes shift of the two compounds before and after the reaction is more than 120 nm, and the cross region of the spectra is less, so the detection results of the designed probe are more reliable and accurate. Based on the oxidation of Tsuji-Trost allyl, a fluorescent probe OHBT. for Pd (0) was designed. When OHBT reacts with Pd (0), the maximum emission wavelength shifts from 410 nm red to 550 nm, with high selectivity to Pd (0). At the same time, Pd (0) was detected by filter paper, and the residual Pd (0) in the coupling product of Suzuki was detected quantitatively.
【学位授予单位】：华东理工大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R346;TP212.2
【参考文献】
相关期刊论文 前3条
1 郭阳雪;李向平;刘桂琴;郑加金;刘靖疆;张桂兰;汤国庆;陈文驹;;7-羟基喹啉在二甲基亚砜中的荧光光谱[J];光谱学与光谱分析;2005年12期
2 肖玉梅;吴燕华;刘吉平;李艳芳;李楠;覃兆海;;β-环糊精及其衍生物对杀菌剂醚菌酯的分子识别作用的研究[J];光谱学与光谱分析;2008年10期
3 吕凤婷,高莉宁,房喻;基于激发态分子内质子转移的新一代荧光探针[J];化学进展;2005年05期
相关博士学位论文 前1条
1 徐兆超;基于ICT萘酰亚胺阳离子比率荧光探针的研究[D];大连理工大学;2006年
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