反應激活型疾病診斷熒光探針及其它相關探針的設計、合成及應用
發(fā)布時間：2018-12-13 01:45
【摘要】：本文設計、合成了三類基于反應激活的熒光探針,反應前后熒光團的電子云密度得到重新排列,使得反應前后可以產生兩個不同波段的熒光發(fā)射峰,因此可以對檢測對象實現比例檢測,大大提高了檢測的精度和可信度。 1.設計、合成了N-乙酰轉移酶II特異性熒光探針Amonafide。臨床代謝研究發(fā)現,化合物amonafide在體內主要被N-乙�；D移酶II催化代謝生成Acetyl-Amonafide�；诖�,設計了化合物Amonafide作為N-乙酰轉移酶II特異性熒光探針�；衔顰monafide和Acetyl-Amonafide在350 nm有共同的最大吸收峰,而發(fā)射波長分別在460 nm和580 nm,更重要的是代謝產物Acetyl-Amonafide的熒光量子產率高達0.74。酶水平和細胞水平實驗進一步證明了探針Amonafide對NAT2的高選擇性和靈敏性(1 nM)。并且成功地將探針Amonafide應用于HepG2細胞株內的N-乙酰轉移酶II熒光成像檢測,這為芳胺類藥物的個性化給藥奠定了基礎。同時完成了分別針對N-乙酰轉移酶I和N-乙酰轉移酶II的長波長熒光探針的設計和部分合成工作。 2.以4-氨基-1,8-萘酰亞胺為熒光團母體,通過異氰酸酯鍵與受體對硝基芐醇鏈接,設計了針對腫瘤乏氧的比率型熒光探針RHP。通過ICT原理,可以使反應后的最大熒光發(fā)射波長紅移(475nm到550 nm)。詳細研究了探針RHP對硝基還原酶(NTR)的熒光響應和動力學曲線,同時檢測了內源性還原物質對探針的影響。實驗結果進一步證明了探針RHP對硝基還原酶的高選擇性。乏氧／有氧實驗證明,當GrHy/GrAr和B1Hy/B1Ar的比值在45以上時,細胞即處于乏氧環(huán)境。探針RHP可以作為腫瘤乏氧診斷探針用于體內實體瘤的早期診斷。同時通過乏氧原理設計并合成了基于Amonafide的前藥化合物。 3.以苯并噻唑類化合物(HBTBC)為熒光團母體,設計了一類基于ESIPT的熒光探針。反應前后兩種化合物的斯托克斯位移高達120 nm以上,且光譜交叉的區(qū)域少,設計的探針檢測結果的可信度和準確度更高。以Tsuji-Trost烯丙基的氧化反應為基礎設計了針對Pd(0)的熒光探針OHBT。當OHBT與Pd(0)反應后,最大發(fā)射波長由410 nm紅移到550 nm,設計的探針對Pd(0)有著高度的選擇性。同時通過濾紙對Pd(0)實現了固體熒光檢測,并成功對Suzuki偶聯反應產物中殘留的Pd(0)進行了定量檢測。
[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.
【學位授予單位】：華東理工大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R346;TP212.2
本文編號：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.
【學位授予單位】：華東理工大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R346;TP212.2
【參考文獻】
相關期刊論文 前3條
1 郭陽雪;李向平;劉桂琴;鄭加金;劉靖疆;張桂蘭;湯國慶;陳文駒;;7-羥基喹啉在二甲基亞砜中的熒光光譜[J];光譜學與光譜分析;2005年12期
2 肖玉梅;吳燕華;劉吉平;李艷芳;李楠;覃兆海;;β-環(huán)糊精及其衍生物對殺菌劑醚菌酯的分子識別作用的研究[J];光譜學與光譜分析;2008年10期
3 呂鳳婷,高莉寧,房喻;基于激發(fā)態(tài)分子內質子轉移的新一代熒光探針[J];化學進展;2005年05期
相關博士學位論文 前1條
1 徐兆超;基于ICT萘酰亞胺陽離子比率熒光探針的研究[D];大連理工大學;2006年
，本文編號：2375645
本文鏈接：http://sikaile.net/xiyixuelunwen/2375645.html
最近更新
教材專著
