基于罗丹明和BODIPY单元的FRET探针的合成及其对小分子α-酮戊二酸的检测

doi:10.7517/j.issn.1674-0475.2017.03.265

摘要/Abstract

摘要：

小分子α-酮戊二酸（α-KA）是人体新陈代谢的一种十分重要的产物，与多种疾病存在直接或者间接的关系，可以对临床诊断提供帮助。以BODIPY单元作为能量给体、罗丹明单元作为能量受体，利用二者存在重叠的光谱，本文设计了一种基于罗丹明和BODIPY单元的荧光共振能量转移（FRET）探针，通过将罗丹明与BODIPY的衍生物进行桥联，得到能够检测α-酮戊二酸的荧光比率型探针RDM-BO。研究表明，探针RDM-BO本身的荧光光谱与BODIPY单元类似，当探针与α-酮戊二酸发生反应时，RDM-BO中的罗丹明单元会发生开环反应，使分子的共轭发生变化，同时产生新的荧光峰，检测效果显著，检测限可达到3.14 μmol·L^-1。

关键词: 罗丹明, BODIPY, &alpha, -酮戊二酸, 荧光探针

Abstract:

Small molecule α-ketoglutaric acid (α-KA) is a very important product of metabolism in human body, which exhibits direct or indirect relation with many kinds of diseases. A fluorescence resonance energy transfer (FRET) ratiometric probe for α-KA containging rhodamine and BODIPY has been synthesized. As the energy donor, the fluorescence spectrum of BODIPY matches well with the absorption spectrum of rhodamine which is chosen as the energy acceptor. Research shows that the orginal fluorescence spectrum of probe RDM-BO is similar with BODIPY. While the α-KA induces ring-open reacntion with rhodamine unit of RDM-BO, the molecular conjugate is changed, resulting in new fluorescence peaks. The detection limit could reach 3.14 μmol·L^-1.

Key words: rhodamine, BODIPY, α-ketoglutaric acid, fluroescence probe

何业, 唐俊马, 郭志前, 靳鹏伟, 朱世琴, 朱为宏. 基于罗丹明和BODIPY单元的FRET探针的合成及其对小分子α-酮戊二酸的检测[J]. 影像科学与光化学, 2017, 35(3): 265-273.

HE Ye, TANG Junma, GUO Zhiqian, JIN Pengwei, ZHU Shiqin, ZHU Weihong. A FRET Fluorescent Probe for α-Ketoglutaric Acid Based on Rhodamine and BODIPY[J]. Imaging Science and Photochemistry, 2017, 35(3): 265-273.

参考文献

[1] Yang Y M, Zhao Q, Feng W, Li F Y. Luminescent chemodosimeters for bioimaging[J]. Chemical Reviews, 2013, 113(1): 192-270.
[2] Virginie D, Francis F, Anthony W C. A Long-wavelength fluorescent chemodosimeter selective for Cu(Ⅱ) ion in water[J]. Journal of the American Chemical Society, 1997, 119(31): 7386-7387.
[3] 韩文, 杨运旭, 许太林, 盛瑞隆, 李斌莲, 朱海波. 一种罗丹明-氨乙基邻苯二甲酰胺衍生物的设计合成及其对Cr³⁺离子的荧光-比色检测[J]. 影像科学与光化学, 2015, 33(2): 144-153. Han W, Yang Y X, Xu T L, Sheng R L, Li B L, Zhu H B. Rhodamine-pathalic diamide dereivative for colorimetric and fluorescent "turn-on" detection of Cr³⁺[J]. Imaging Science and Photochemistry, 2015, 33(2): 144-153.
[4] Chen X, Pradhan T, Wang F, Kim J S, Yoon J. Fluorescent chemosensors based on spiroring-opening of xanthenes and related derivatives[J]. Chemical Reviews, 2012, 112(3): 1910-1956.
[5] Zhang X L, Xiao Y, Qian X H. A ratiometric fluorescent probe based on fret for imaging Hg²⁺ ions in living cells[J]. Angewandte Chemie International Edition, 2008, 47(42): 8025-8029.
[6] Hippius C, Schlosser, Vysotsky M O, Böhmer V, Würthner F. Energy transfer in calixarene-based cofacial-positioned perylene bisimide arrays[J]. Journal of the American Chemical Society, 2006, 128(12): 3870-3871.
[7] 杨博, 吴文辉. 具有荧光增强性能的罗丹明B衍生物的合成及其对汞离子的检测[J]. 影像科学与光化学, 2013, 31(6): 421-429. Yang B, Wu W H. Synthesis of the rhodamine B derivatives with fluorescence "turn-on" effect and their application to the detection of Hg²⁺[J]. Imaging Science and Photoche-mistry, 2013, 31(6): 421-429.
[8] Nierth A, Kobitski A Y, Nienhaus G U, Jäschke A. Anthracene-BODIPY dyads as fluorescent sensors for biocatalytic diels-alder reactions[J]. Journal of the American Chemical Society, 2010, 132(8): 2646-2654.
[9] Zadran S, Remacle F, Levine R D. Reversible photochromic system based on rhodamine B salicylaldehyde hydrazone metal complex[J]. Journal of the American Chemical Society, 2014, 136(4): 1643-1649.
[10] Niu L Y, Guan Y S, Chen Y Z, Wu L Z, Tung C H, Yang Q Z. BODIPY-based ratiometric fluorescent sensor for highly selective detection of glutathione over cysteine and homocysteine[J]. Journal of the American Chemical Society, 2012, 134(46): 18928-18931.
[11] Dinardo C D, Propert K J, Loren A W, Paietta E, Sun Z X, Levine R L, Straley K S, Yen K, Patel J P, Agresta S, Abdel-Wahab O, Perl A E, Litzow M R, Rowe J M, Lazarus H M, Fernandez H F, Margolis D J, Tallman M S, Luger S M, Carroll M. Serum 2-hydroxyglutarate levels predict isocitrate dehydrogenase mutations and clinical outcome in acute myeloid leukemia[J]. Blood, 2012, 121(24): 4917-4924.
[12] Zadran S, Remacle F, Levine R D. miRNA and mRNA cancer signatures determined by analysis of expression levels in large cohorts of patients[J]. Proceedings of the National Academy of Sciences, 2013, 110(47): 19160-19165.
[13] Masato S, Christiane B K, Joshua C M, Evan F L, Dirk B, Anne B, Isaac S H, Roxanne H, Andrew W, Jillian H, Annick Y, Wanda Y L, Stefanie S, Shinsan M S, Carl V, Guido R, Pamela S O, Dwayne L. B, Maria E. F, Ari M, Juan-Carlos Z P, Tak W M. IDH1(R132H) mutation increases murine haematopoietic progenitors and alters epigenetics[J]. Nature, 2012, 488: 656-659.
[14] Gallego E R, Guirro M, Riera-Borrull M, Aguilera A H, Casadó R M, Arroyo S F, Debon R B, Sabench F, Hernandez M, Castillo D, Menendez J A, Camps J, Ras R, Arola L, Joven J. Mapping of the circulating metabolome reveals α-ketoglutarate as a predictor of morbid obesity-associated non-alcoholic fatty liver disease[J]. International Journal of Obesity, 2015, 39: 279-287.
[15] Langenbeck U, Möhring H U, Dieckmann K P. Gas chromatography of α-keto acids as their o-trimethylsilylquino-xalinol derivatives[J]. Journal of Chromatography A, 1975, 115(1): 65-70.
[16] Montenegro P, Valente I M, Goncalves L M, Rodrigues J A, Barros A A. Single determination of α-ketoglutaric acid and pyruvic acid in beer by HPLC with UV detection[J]. Analytical Methods, 2011, 3(5): 1207-1212.
[17] Jin P W, Jiao C H, Guo Z Q, He Y, Zhu S Q, Tian H, Zhu W H. Rational design of a turn-on fluorescent sensor for α-ketoglutaric acid in a microfluidic chip[J]. Chemical Science, 2014, 5(10): 4012-4016.
[18] Cai Q, Yu T, Zhu W P, Xu Y F, Qian X H. A turn-on fluorescent probe for tumor hypoxia imaging in living cells[J]. Chemical Communications, 2015, 51(79): 14739-14741.
[19] Sakabe M, Asanuma D, Kamiya M R, Iwatate J, Hanaoka K, Terai T, Nagano T, Urano Y. Rational design of highly sensitive fluorescence probes for protease and glycosidase based on precisely controlled spirocyclization[J]. Journal of the American Chemical Society, 2013, 135(1): 409-414.
[20] Karaku? E, Üçüncüa M, Emrullaho?lu M. A rhodamine/BODIPY-based fluorescent probe for the dierential detection of Hg(Ⅱ) and Au(Ⅲ)[J]. Chemical Communications, 2014, 50(9): 1119-1121.
[21] Zhou L Y, Zhang X B, Wang Q Q, Lv Y F, Mao G J, Luo A L, Wu Y X, Wu Y, Zhang J, Tan W H. Molecular engineering of a FRET-based two-photon fluorescent probe for ratiometric imaging of living cells and tissues[J]. Journal of the American Chemical Society, 2014, 136(28): 9838-9841.
[22] Sakabe M, Asanuma D, Kamiya M, Iwatate R J, Hanaoka K, Terai T, Nagano T, Urano Y. Rational design of highly sensitive fluorescence probes for protease and glycosidase based on precisely controlled spirocyclization[J]. Journal of the American Chemical Society, 2013, 135(1): 409-414.
[23] Sekiya M, Umezawa K, Sato A, Citterio D, Suzuki K. A novel luciferin-based bright chemiluminescent probe for the detection of reactive oxygen species[J]. Chemical Comm-unications, 2009, 21: 3047-3049.
[24] Yu H B, Xiao Y, Guo H Y. From spirolactam mixtures to regioisomerically pure 5-and 6-rhodamines: a chemodosi-meter inspired strategy[J]. Organic Letters, 2012, 14(8): 2014-2017.
[25] 焦长红,何业, 靳鹏伟,朱世琴,朱为宏. 基于罗丹明B的激活型α-酮戊二酸荧光探针[J]. 影像科学与光化学, 2015, 33(3): 195-202. Jiao C H, He Y, Jin P W, Zhu S Q, Zhu W H. A turn-on fluorescent probe for α-ketoglutaric acid based on rhodamine B[J]. Imaging Science and Photochemistry, 2015, 33(3): 195-202.
[26] Shortreed M, Kopelman R, Kuhn M, Hoyland B. Fluorescent Fiber-optic calcium sensor for physiological measurements[J]. Analytical Chemistry, 1996, 68(8): 1414-1418.