A New Diketopyrrolopyrrole-based Ratiometric Fluorescent Probe for

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A New Diketopyrrolopyrrole-based Ratiometric Fluorescent Probe for Intracellular Esterase Detection and Discrimination of Live and Dead Cells in Different Fluorescence Channel Jian Wang, Weibo Xu, Zhicheng Yang, Yongchao Yan, Xiaoxu Xie, Ning Qu, Yu Wang, Chengyun Wang, and Jianli Hua ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.8b11365 • Publication Date (Web): 21 Aug 2018 Downloaded from http://pubs.acs.org on August 22, 2018

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ACS Applied Materials & Interfaces

A New Diketopyrrolopyrrole-based Ratiometric Fluorescent Probe for Intracellular Esterase Detection and Discrimination of Live and Dead Cells in Different Fluorescence Channel Jian Wanga,§, Weibo Xub,c,§, Zhicheng Yanga, Yongchao Yana, Xiaoxu Xiea, Ning Qub,c, Yu Wangb,c*, Chengyun Wanga, Jianli Huaa* a

Key Laboratory for Advanced Materials & Institute of Fine Chemicals, College of Chemistry and Molecular Engineering, East China University of Science and Technology, 130 Meilong Rd., Shanghai 200237, China b

Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China.

c

Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, China.

Abstract: A new diketopyrrolopyrrole-based fluorescent probe (DPP-AM) was designed and synthesized for ratiometric detection of esterase and imaging of live and dead cells in different modes. DPP-AM showed red fluorescence because of the intramolecular charge transfer (ICT) progress from DPP moiety to pyridinium cation, thus gave remarkable ratio changes (about 70 folds) with fluorescence changed from red to yellow after treating with esterase due to the broken ICT progress. Besides, the detection limit of DPP-AM towards esterase in vitro was 9.51×10-5 U/mL. After pretreated by H2O2 and UV light radiation, the health status of TPC1 cells has been successfully imaged. More importantly, DPP-AM showed yellow fluorescence in live cells and red fluorescent signal in dead cells, indicating that DPP-AM has great potential applications for assessing esterase activity as well as discriminating live and dead cells. KEYWORDS: diketopyrrolopyrrole, fluorescent probe, ratiometric, esterase, live and dead cells fluorescent probes have been reported to sensing and imaging of esterase activity in live cells based on “turn-on” fluorescence sensing systems.19-22 Due to higher detecting accuracy and larger change of ratio signal, ratiometric fluorescent probes often show higher sensitivity.23-25 Therefore, few ratiometric fluorescent probes based on esterase activities have been reported for live cell imaging.26,27 Recently, an unprecedented fluorescent probe was reported for discrimination of live and dead cells by utilizing the presence and absence of esterase activity,28 which further demonstrate the potential significant applications of ratiometric fluorescent probes toward esterase in biosystems. Diketopyrrolopyrrole (DPP) and their derivatives, generally show excellent photothermal stability, good photophysical properties, long absorption and emission wavelength, and high fluorescence quantum yield, thus has a very widely applications in different fields, such as organic photovoltaics,29-31 two-photon absorption materials,32,33 semiconductors34,35 and photodynamic and photothermal therapy materials.36,37 Without any modification, DPP cores showed a fluorescence emission exceeded 500 nm with strong yellow fluorescence, and easily red shift to red fluorescence via appropriate modification as a long-wavelength fluorophore to design fluorescent probes, which make it potential for sensing and bioimaging.38-44

Introduction Esterases, are a group of enzymes which were widely found in various organisms tissue cells that mediate enzyme-catalyzed hydrolysis of various esters,1,2 and involved in various metabolic functions such as ester metabolism,3,4 protein metabolism,5,6 detoxification,7,8 and signal transmission.9,10 For example, insufficiency of lysosomal esterase would bring about wolman disease accompanied by various symptoms including abdominal swelling, diarrhea, hepatomegaly and failure to gain weight.11-13 Furthermore, the design, delivery and release of some prodrugs are also associated with intracellular esterase activity.14-16 More importantly, as one of the major determining factor of the metabolism in cells, esterase play a significant role in cell viability and cytotoxicity assays.17,18 Consequently, developing a reliable assay method is favorable for the quantitation of esterase activity as well as the analysis of cellular status. Fluorescent probes, are those molecules possess fluorescence characteristics, such as fluorescence lifetime, wavelength, and emission intensity, change as a result of specific reactions with target molecules, provided an essential real tool for real-time detection and visualization of some biological species due to its high temporal, non-invasiveness, high sensitivity and spatial resolution, thus become more and more attractive. Up to now, various small molecule-based

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well confirmed by 1H NMR, (Supporting Information).

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C NMR, and HRMS

Spectroscopic Response of DPP-AM to Esterase. The photoluminescence and UV-vis properties of probe DPP-AM toward esterase were studied in DMSO-PBS solution (2:8, v/v, pH = 7.4). Firstly, time-dependent fluorescence response toward 0.1 U/mL esterase from 0 to 24 min was conducted (Figure S1), which demonstrated that the reaction could almost completed within 20 min. As shown in Figure 1, probe DPP-AM itself showed a maximum absorption peak at 520 nm. Upon addition of 0.1 U/mL esterase, the maximum absorption peak of DPP-AM was slightly blue-shifted to 500 nm, which can be observed by naked eye. Meanwhile, the fluorescence spectra of DPP-AM featured an obvious red fluorescence with maximum emission at 655 nm (Φf = 0.41) in the absence of esterase. However, the fluorescence intensity at 655 nm decreased gradually and a new distinct blue-shifted fluorescent peak at 551 nm (Φf = 0.67) emerged with addition of esterase from 0 to 0.1 U/mL, affording a significant ratiometric fluorescent signal (I551/I655). This phenomenon implied that esterase can trigger cleavage of acetoxyl of DPPAM to remove pyridinium cation, thus broken the ICT process from the DPP moiety to pyridine. Moreover, the fluorescence ratio I551/I655 increased by 70-fold upon reaction with 0.1 U/mL esterase, implying that DPP-AM was favorable for the ratiometric fluorescence determination of esterase.

Scheme 1. Chemical structure of DPP-AM and proposed reaction mechanisms of DPP-AM toward esterase. In this work, a new ratiometric fluorescent probe was designed and synthesized (DPP-AM, Scheme 1). The probe was constructed by introducing 4-bromomethyl-phenyl acetate to pyridine-cotained DPP core. The formation of pyridinium cation made DPP-AM show strong red fluorescence with maximum emission peak at about 655 nm by intramolecular charge transfer (ICT) process. Upon reaction with esterase, however, the cation disappeared because acetyl ester bond is cleaved by the enzymatic hydrolysis, thus broken the ICT process.45,46 As a result, the emission peak was blue-shifted to 551 nm with yellow fluorescence emerged, which made DPPAM as a suitable ratiometric fluorescent probe for imaging of esterase activity. In addition, DPP-AM was successfully applied to evaluate the healthy status of cells and discriminate live and dead cells, which based on the high esterase activity in live cells 47,48 and the insufficient esterase activity in injured or dead cells.49,50

Results and Discussion

Figure 1. (A) UV-vis absorption spectra of DPP-AM (10 µM) before and after addition of 0.1 U/mL esterase in DMSO-PBS solution (DMSO : PBS = 2:8, v/v, pH = 7.4) at 37 oC for 20 min. Insert: photograph showing the visual color of DPP-AM when treated without (left) or with (right) esterase under visible light. (B) Fluorescence emission spectra of DPP-AM (10 µM) with 0-0.1 U/mL esterase at 37 oC for 20 min. Insert: photograph showing the visual fluorescence color of DPP-AM when treated without (left) or with (right) easterase under a 365 nm UV lamp. λex = 490 nm.

Synthesis of DPP-AM The synthetic route of target compound (DPP-AM) was shown in Scheme 2. Firstly, compound 1 and 2 were synthesized according to our previous work,51 respectively. In the next step, compound 3 was prepared through cyclization of compound 2 and 4cyanopyridine in the presence of sodium tert-pentoxide. Then, the reaction of compound 3 with 1-bromohexane provided important intermediate 4 in the catalysis of sodium hydride. Finally, the desired product DPP-AM was obtained by a nucleophilic substitution reaction between compound 4 and 4bromomethylphenyl acetate. All the new compounds were

Considering of the analytical conditions, the effects of pH and reaction temperature on the fluorescence of DPP-AM were examined. As shown in Figure S2 (Supporting Information), the fluorescence ratio of DPP-AM was almost impervious to the change of pH from 6.5 to 9.0 in DMSO-PBS solution and temperature from 30 to 47 °C, and it reached its maximum value at about pH 7.4 and 37 °C after reaction with 0.1 U/mL esterase. Consequently, the reaction of DPP-AM with esterase can performed best under physiological conditions (pH = 7.4, 37 °C).

Scheme 2. The synthesis of the target DPP-AM.


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ACS Applied Materials & Interfaces Study of esterase activities in solution. The ratio fluorescence kinetic curves of DPP-AM upon incubation with esterase at varied concentrations were shown in Figure 2. As shown in Figure 2A, faster cleavage and stronger fluorescence ratio increasement as higher concentration of esterase added (0-0.08 U/mL). In contrast, the fluorescence of DPP-AM almost has no changed without treated with esterase during the same period of time (Figure 2A, black line), which showed that DPP-AM has good stability in the detection system.

including inorganic salts (KCl, Na2SO4, Na2CO3), reactive oxygen species (ClO-, H2O2), three common amino acids (Cys, Hcy, GSH), ATP, ADP and proteins (HSA, BSA, AFP, LAP, APN). As shown in Figure 3B, the esterase can also induce about 43-53 fold ratiometric change though different interference substances exist, which demonstrates that DPPAM has outstanding selectivity towards esterase over other common biological species.

According to the determined condition aboved, the fluorescence response of DPP-AM exhibited a good linearity with easterase (Figure 3A) in the concentration range of 00.08 U/mL with an equation of R = 48.63 × C (U/mL) + 0.19 (R = 0.991), where R is the fluorescence ratio (I551/I655) of DPP-AM after addition of easterase with diffirent concentrations. The detection limit (3δ/k) is determined to be 9.51×10-5 U/mL easterase.

Figure 3. (A) The linear fitting curve of R towards the concentration of esterase between 0-0.08 U/mL. λex = 490 nm. (B) Fluorescence responses of DPP-AM (10 µM) to various species: Na2CO3 (100 µM), Na2SO4 (100 µM), KCl (100 µM), NaClO (100 µM), H2O2 (100 µM), Hcy (50 µM), Cys (50 µM), GSH (50 µM), ATP (50 µM), ADP (50 µM), HSA (50 µM), BSA (50 µM), AFP (10.0 ng/mL), LAP (10.0 ng/mL), APN (10.0 ng/mL) and esterase (0.1 U/mL) in DMSO-PBS buffer solution (DMSO : PBS = 2 : 8, v:v, pH = 7.4, 37 oC) with λex = 490 nm.

Figure 2. (A) Plot of fluorescence emission ratio (I551/I655) of DPP-AM (10 µM) vs. the reaction time at different esterase concentrations (from bottom to top): 0 (control), 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07 and 0.08 U/mL. (B) The fluorescence ratio (I551/I655) of DPP-AM (10 µM) upon incubation with esterase (0.08 U/mL) as a function of time in the presence of various concentrations of AEBSF (from top to bottom): 0, 0.20, 0.40, 0.60, 0.80 and 1.0 mM.

Enzyme-Catalytic Activation Mechanism. As mentioned above, the ratiometric fluorescence changes were attributed to the cleavage of acetoxyl by the enzymetriggered reaction, thus produced the weaken electro-withdrawing pyridine group accompanying with the generation of compound 4. To undersatnd the interaction mechanism, high performance liquid chromatography (HPLC) measurements was used for compound 4 as well as DPP-AM before and after reation with esterase. As illustrated in Figure 4, compound 4 and DPPAM exhibited a chromatographic peak with retention time at 23.7 and 12.2 min, respectively. While addition of the reaction mixture of DPP-AM and esterase, a chromatographic peak at 23.7 min which is corresponding to compound 4 could be clearly observed, a weak peak was also observed at 12.2 min, corresponding to probe DPP-AM. Moreover, the product of the mixture of DPP-AM and esterase was further investigated by HRMS. DPP-AM showed a peak at m/z 606.3329 [M]+, After reaction with esterase, a peak at m/z 458.2802 [M + H]+, which is corresponding to compound 4 could be found (Figure 4D), indicating the formation of compound 4 upon the enzymereactive cleavage reaction. These data demonstrated that DPP-AM could react with esterase to liberate compound 4, which achieved the fluorescence transformation from red to yellow.

To further validate that the ratio (I551/I655) change was caused by esterase, 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), a inhibitor of esterase, was investigated the inhibition effect on esterase activity. As depicted in Figure 2B, as AEBSF concentration increased, the fluorescence ratio (I551/I655) of DPP-AM decreased gradually. For example, when 0.2 mM AEBSF was added, 23.8% decrease of the original fluorescence ratio was detected. When more AEBSF such as 0.6 and 1.0 mM were added, a larger decrease (60.74% and 87.75%) of the original fluorescence ratio was occurred, respectively (Figure S3). All these results further demonstrate that the ratiometric fluorescence response of DPP-AM toward esterase indeed arises from the enzymatic hydrolysis. The selectivity of DPP-AM towards esterase was then investigated by measuring its emission intensity ratio (I551/I655) in the presence of different interference substances (Figure 3),



Figure 5. Fluorescent imaging of TPC1 cells stained with 10 µM of DPP-AM with different incubation times. Incubation time: (A, F, K) 5 min; (B, G, L) 15 min; (C, H, M) 30 min; (D, I, N) 45 min; (C, J, O) 60 min. (A-E) yellow channel (525 nm-585 nm); (F-J) red channel (625 nm-685 nm); (K-O) merge image. λex = 490 nm. Scale bar = 10 µm. yellow fluorescence in the range of 525 nm-585 nm, rather than almost invisible red emission in 625 nm-685 nm. Meanwhile, as shown in Figure 6D-F, yellow channel signal was blocked and a remarkable red fluorescence was observed after pretreated cells with AEBSF (0.8 mM) for 60 min, these signal changes demonstrated that DPP-AM had specific response towards esterase in living cells. Figure 4. HPLC analysis of (A) DPP-AM, (B) the reaction product of DPP-AM by esterase and (C) compound 4. (D) HRMS of DPP-AM and the reaction product of DPP-AM by esterase. Cell Imaging. With the photophysical properties of DPPAM in evaluating the esterase activity in vitro, the applications of the probe in living samples was further examined. Firstly, the cytotoxicity of DPP-AM was evaluated by CCK-8 assay. As shown in Figure S4, DPP-AM showed low cytotoxicity towards TPC1 cell with over 80% cell viabilities after 24hours incubation at a concentration of 10 µM, which indicated that DPP-AM was suitable for the applications in biosystems. Then, in order to understand the optimum incubation time of DPP-AM in live cells, cell images of DPP-AM (10 µM) incubation with cells in different times were collected. As depicted in Figure 5, slight fluorescence in yellow channel became occurred after incubation 15 min, and fluorescence decreased in red channel was observed. As the incubation time up to 30 min, a significant fluorescence enhancement in yellow channel was observed, and reached the plateau point after about 45 min incubation. Furthermore, DPP-AM incubated with normal cell (293T) for different times was also investigated, which gave similar changes in yellow and red channels (Figure S5). Herein, the fluorescence changes of DPP-AM can trigger by intracellular esterase, thus indicated DPP-AM was suitable for esterase activity monitoring in living cells.

Figure 6. Fluorescent images of live (A-C) and dead (G-I) TPC1 cells stained with DPP-AM (10 µM) for 45 min and (DF) pretreated TPC1 cells with AEBSF (0.8 mM) for 60 min and then further incubated with DPP-AM (10 µM) for 45 min. Yellow channel (525 nm-585 nm), red channel (625 nm-685 nm). λex = 490 nm. Scale bar = 10 µm. Based on these, dead cells, which obtained by treated live cells with paraformaldehyde, gave obvious red fluorescence in 625 nm-685 nm, instead of weak yellow emission in the range of 525 nm-585 nm because of lacking active eserase. All these results revealed that DPP-AM could be used for

Subsequently, imaging of DPP-AM in live and dead cells were conducted. As displayed in Figure 6, after incubated with DPP-AM (10 µM) for 45 min, live cells showed strong


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ACS Applied Materials & Interfaces discrimination of live and dead cells in yellow and red two fluorescent channels. Strong yellow channel signal implies cells are rigorous, while once cells are unhealthy or death, red fluorescence could be detected.

unhealthy even death (Figure S4C). To verify if DPP-AM has the same ability to detect cell unhealthy status pretreated by UV radiation vs H2O2, TPC1 cells were pretreated by different exposure time under 365 nm UV light and then incubation with DPP-AM for 45 min. As shown in Figure 8A, TPC1 cells without UV radiation showed strong emission signal in yellow channel, while almost no signal could be detected in red channel. With the increase of UV radiation time, the yellow fluorescence decreased gradually (Figure 8B1-E1), and the fluorescence in red channel turned stronger (Figure 8B2-E2), demonstrating that DPP-AM has the capability to analyze the degree of damage caused by UV radiation as well as unhealthy status of TPC1 cells.

Analysis of Healthy Status of Cells Pretreated with H2O2. Hydrogen peroxide (H2O2) is a byproduct produced through cellular respiration and metabolism in the human body. It is believed that H2O2 belongs to reactive oxygen species (ROS), which has significant destructive effect on cells and tissues, thus make cells injured even death (Fifure S4B). As shown in Figure S6, the maximum absorbance of DPP-AM and compound 4 showed no more than 13% and 10% reduction respectively after addition of 20 mM H2O2 and performed excellent antioxidant capacity, indicating that H2O2 has ignorable influence towards these two compounds. Therefore, TPC1 cells were pretreated with different amount of H2O2 before incubation with DPP-AM (10 µM), in order to test if probe DPP-AM could reveal the health status of cells. As shown in Figure 7A, cells without pretreated with H2O2 showed significant yellow emission signal in yellow channel, while almost no signal could be detected in red channel, indicating that highly active esterase in cells, thus demonstrated almost no damages to the status of cells. As shown in Figure 7, with the pretreatment concentration of H2O2 increased, the yellow fluorescence turns weaker (Figure 7B1-E1), while the emission signals in red channel increased gradually (Figure 7B2-E2), demonstrating the restricted esterase activity because of cell activity decreased gradually after H2O2 stimulated. Therefore, DPP-AM can be used to evaluate the different status of cells.

Figure 8. Fluorescent images of TPC1 cells pretreated with UV exposure for different time and then incubated with 10 µM of DPP-AM for 45 min. Exposure time: (A) 0 min; (B) 10 min; (C) 25 min; (D) 40 min; (E) 60 min. (A1-E1) yellow channel (525 nm-585 nm); (A2-E2) red channel (625 nm-685 nm); (A3-E3) merge image; (A4-E4) bright field. λex = 490 nm. Scale bar = 10 µm.

Conclusions In summary, a new ratiometric fluorescent probe DPP-AM for esterase detection and imaging has been developed. DPP-AM showed specific recognition towards esterase in the presence of other analytes. To our delight, DPP-AM was successfully applied to monitor esterase activity both in cancer and normal living cells. Furthermore, DPP-AM can be converted to compound 4 under the catalysis of active esterase, thus showed fluorescence signal in 525 nm-585 nm in living cells. While DPP-AM exhibited red emission in 625 nm-685 nm in dead cells due to the absence of esterase. Accordingly, DPPAM can discriminate live and dead cells based on yellow and red fluorescence signal and show more accurate definition compared to those probes only stain live or dead cells in single channel. Finally, the probe was uesd to analyze the cell living status after pretreated with H2O2 and UV radiation. The results provide a strategy to design more ratiometric fluorescent probes for esterase activity monitor, as well as a potential powerful tool for biological applications.

Figure 7. Fluorescent images of TPC1 cells incubated with 10 µM of DPP-AM for 45 min after pretreated with different amounts of H2O2 for 2 h. H2O2 concentration: (A) 0 mM; (B) 0.5 mM; (C) 1.0 mM; (D) 4.0 mM; (E) 8.0 mM. (A1-E1) yellow channel (525 nm-585 nm); (A2-E2) red channel (625 nm-685 nm); (A3-E3) merge image; (A4-E4) bright field. λex = 490 nm. Scale bar = 10 µm. Analysis of Cell Unhealthy Status Induced by UV Radiation. Ultraviolet (UV) light, which wavelength is under 400 nm, may cause DNA mutation in cells, thus make cells



8.

Supporting Information Available: Supporting Figures (Figure S1-S6) as mentioned in the main text in the manuscript are available online. This material is available free of charge via Internet at http://pubs.acs.org.

9.

Figures showing procedure, instruments and methods on the cell imaging experiments, effects of the system pH and temperature, time-dependent emission spectra of DPP-AM, stability of DPP-AM to H2O2 and light irradiation, cytotoxicity of DPP-AM, fluorescence images of 293T cells treated with DPP-AM, and original spectral copy of new compounds.

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AUTHOR INFORMATION 12.

Corresponding Author * E-mail: [email protected]. Tel: (+86) 21 64250940 * E-mail: [email protected].

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Author Contributions §

These authors contributed equally.

14.

Notes The authors declare no competing interests.

Acknowledgment

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This work was supported by the National Natural Science Foundation of China (21772040, 21421004, 21372082 and 21572062), the Fundamental Research Funds for the Central Universities (222201717003) and the Programme of Introducing Talents of Discipline to Universities (B16017).

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References 1.

2.

3.

4.

5.

6.

7.

17.

Redinbo, M. R.; Potter, P. M. Mammalian Carboxylesterases: From Drug Targets to Protein Therapeutics. Drug Discov Today 2005, 10, 313−325. Rautio, J.; Kumpulainene, H.; Heimbach, T.; Oliyai, R.; Oh, D.; Jarvinen, T.; Savolainen, J. Nat. Rev. Drug Discov 2008, 7, 255−270. Mutch, E.; Nave, R.; McCracken, N.; Zech, K.; Williams, F. M. The Role of Esterases in the Metabolism of Ciclesonide to Desisobutyryl-Ciclesonide in Human Tissue. Biochem Pharmacol 2007, 73, 1657−1664. Yoon, K. J. P.; Morton, C. L.; Potter, P. M.; Danksa, M. K.; Lee, R. E. Synthesis and Evaluation of Esters and Carbamates to Identify Critical Functional Groups for Esterase-specific Metabolism. Bioorg. Med. Chem. 2003, 11, 3237−3244. Yang, F.; Bian, C. B.; Zhu, L. L.; Zhao, G. X.; Huang, Z. X.; Huang, M. D. Effect of Human Serum Albumin on Drug Metabolism: Structural Evidence of Esterase Activity of Human Serum Albumin. J. Struct Biol. 2007, 157, 348−355. Guénin, S.; Hardouin, J.; Paynel, F.; Müller, K.; Mongelard, G.; Driouich, A.; Lerouge, P.; Kermode, A. R.; Lehner, A.; Mollet, J. C.; Pelloux, J.; Gutierrez, L.; Mareck,A. AtPME3, A Ubiquitous Cell Wall Pectin Methylesterase of Arabidopsis Thaliana, Alters the Metabolism of Cruciferin Seed Storage Proteins During Post-germinative Growth of Seedlings. J. Exp Bot. 2017, 68, 1083−1095. Chen, S. W.; Hsu, J. T.; Chou, Y. A.; Wang, H. T. The Application of Digestive Tract Lactic Acid Bacteria with High Esterase Activity for Zearalenone Detoxification. J. Sci Food Agric. 2018, 98, 3870−3879.

18.

19.

20.

21.

22. 23.

24.

25.

Sanchez-Hernandez, J. C.; Notario del Pino, J.; Domínguez, J. Earthworm-induced Carboxylesterase Activity in Soil: Assessing the Potential for Detoxification and Monitoring Organophosphorus Pesticides. Ecotox. Environ. Safe. 2015, 122, 303−312. Munger J. S.; Shi G. P.; Mark E. A. A Serine Esterase Released by Human Alveolar Macrophages Is Closely Related to Liver Microsomal Carboxylesterases. J Bio. Chem. 1991, 266, 18832−18838. Maeintyre S.; Smols D.; Diley P. Two Carboxylesterases Bind C-reactive Protein within the Endoplasmic Reticulum and Regulate Its Secretion During the Acute Phase Response. J Bio. Chem. 1994, 269, 24496−24503. Anderson, R. A.; Byrum, R. S.; Coates, P. M.; Sando. G. N. Mutations at the Lysosomal Acid Cholesteryl Ester Hydrolase Gene Locus in Wolman Disease. PNAS 1994, 91, 2718−2722. Fasanoa, T.; Pisciotta, L.; Bocchia, L.; Guardamagna, O.; Assandro, P.; Rabacchi, C.; Zanoni, P.; Filocamo, M.; Bertolini, S.; Calandra. S. Lysosomal Lipase Deficiency: Molecular Characterization of Eleven Patients with Wolman or Cholesteryl Ester Storage Disease. Mol. Genet Metab. 2012, 105, 450−456. Richard A. A.; Greta M. B.; John S. P. Lysosomal Acid Lipase Mutations That Determine Phenotype in Wolman and Cholesterol Ester Storage Disease. Mol. Genet Metab. 1999, 68, 333−345. Fernando, I. R.; Ferris, D. P.; Frasconi, M.; Malin, D.; Strekalova, E.; Yilmaz, M. D.; Ambrogio, M. W.; Algaradah, M. M.; Hong, M. P.; Chen, X. Q.; Nassar, M. S.; Botros, Y. Y.; Cryns, V. L.; Stoddart. J. F. Esterase- and pH-responsive Poly(βaminoester)-capped Mesoporous Silica Nanoparticlesfor Drug Delivery. Nanoscale 2015, 7, 7178−7183. John, S.; Thangapandian, S.; Lazar, P.; Son, M.; Park, C.; Lee, K. W. New Insights in the Activation of Human Cholesterol Esterase to Design Potent Anti-Cholesterol Drugs. Mol Divers 2014, 18, 119−131. Ji, C.; Miller, M. J. Chemical Syntheses and in Vitro Antibacterial Activity of Two Desferrioxamine B-ciprofloxacin Conjugates with Potential Esterase and Phosphatase Triggered Drug Release Linkers. Bioorgan Med Chem. 2012, 20, 3828−3836. Dorsey, J.; Yentsch, C. M.; Mayo, S.; McKenna, C. Rapid Analytical Technique for the Assessment of Cell Metabolic Activity in Marine Microalgae. Cytometry 1989, 10, 622−628. Johnson, I.; Spence, M. T. Z. The Molecular Probes Handbook: A Guide to Fluorescent Probes and Labeling Technologies, Molecular Probes by Life Technologies, 2010. Gao, M.; Hu, Q. L.; Feng, G. X.; Tang, B. Z.; Liu, B. A fluorescent Light-up Probe with “AIE + ESIPT” Characteristics for Specific Detection of Lysosomal Esterase. J. Mater. Chem. B. 2014, 2, 3438−3442. Peng, L.; Xu, S. D.; Zheng, X. K.; Cheng,X. M.; Zhang, R. Y.; Liu, J.; Liu, B.; Tong, A. J. Rational Design of a Red-Emissive Fluorophore with AIE and ESIPT Characteristics and Its Application in Light-Up Sensing of Esterase. Anal. Chem. 2017, 89, 3162−3168. Levine, S. R.; Beatty. K. E. Synthesis of a Far-Red Fluorophore and Its Use as an Esterase Probe in Living Cells. Chem. Commun. 2016, 52, 1835−1838. Tallman, K. R.; Beatty. K. E. Far-Red Fluorogenic Probes for Esterase and Lipase Detection. ChemBioChem 2015, 16, 70−75. Pramanik, S.; Bhalla, V.; Kumar, M. HexaphenylbenzeneBased Fluorescent Aggregates for Ratiometric Detection of Cyanide Ions at Nanomolar Level: Set−Reset Memorized Sequential Logic Device. ACS Appl. Mater. Interfaces 2014, 6, 5930−5939. Kim, H. J.; Heo, C. H.; Kim, H. M. Benzimidazole-Based Ratiometric Two-Photon Fluorescent Probes for Acidic pH in Live Cells and Tissues. J. Am. Chem. Soc. 2013, 135, 17969−17977. Liu, J. W.; Luo, Y.; Wang, Y. M.; Duan, L. Y.; Jiang, J. H.; Yu, R. Q. Graphitic Carbon Nitride Nanosheets-Based Ratiometric


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38.

39.

Fluorescent Probe for Highly Sensitive Detection of H2O2 and Glucose. ACS Appl. Mater. Interfaces 2016, 8, 33439−33445. Kim, S. W.; Kim, H. J.; Choi, Y. D.; Kim. Y. M. A New Strategy for Fluorogenic Esterase Probes Displaying Low Levels of Non-specific Hydrolysis. Chem. Eur. J. 2015, 21, 9645−9649. Komatsu, T.; Urano, Y.; Fujikawa, Y.; Kobayashi, T.; Kojima, H.; Terai, T.; Hanaoka. K.; Nagano, T. Development of 2,6Carboxy-Substituted Boron Dipyrromethene (BODIPY) as a Novel Scaffold of Ratiometric Fluorescent Probes for Live Cell Imaging. Chem. Commun. 2009, 45, 7015−7017. Tian, M. G.; Sun, J.; Tang, Y. H.; Dong, B. L.; Lin. W. Y. Discriminating Live and Dead Cells in Dual-Color Mode with a Two Photon Fluorescent Probe Based on ESIPT Mechanism. Anal. Chem. 2018, 90, 998−1005. Qu, S. Y.; Tian, H. Diketopyrrolopyrrole (DPP)-Based Materials for Organic Photovoltaics. Chem. Commun. 2012, 48, 3039−3051. Jung, J. W.; Liu, F.; Russell, T. P.; Jo, W. H. A High Mobility Conjugated Polymer Based on Dithienothiophene and Diketopyrrolopyrrole for Organic Photovoltaics. Energy Environ. Sci. 2012, 5, 6857−6861. Wang, Q.; Franeker, J. J.; Bruijnaers, B. J.; Wienk, M. M.; Janssen, R. A. J. Structure-Property Relationships for BisDiketopyrrolopyrrole Molecules in Organic Photovoltaics. J. Mater. Chem. A. 2016, 4, 10532−10541. Gao, Y. T.; Feng, G.; Jiang, T.; Goh, C.; Ng, L.; Liu, B.; Li, B.; Yang, L.; Hua, J. L.; Tian, H. Biocompatible Nanoparticles Based on Diketo-Pyrrolo-Pyrrole (DPP) with AggregationInduced Red/NIR Emission for in Vivo Two-Photon Fluorescence Imaging. Adv. Func. Mater. 2015, 25, 2857−2866. Ftouni, H.; Bolze, F.; Rocquigny, H.; Nicoud, J. F. Functionalized Two-Photon Absorbing Diketopyrrolopyrrole-Based Fluorophores for Living Cells Fluorescent Microscopy. Bioconjugate Chem. 2013, 24, 942−950. Back, J. Y.; Yu, H.; Song, I.; Kang, I.; Ahn, H.; Shin, T. J.; Kwon, S. K.; Oh, J. H.; Kim, Y. H. Investigation of StructureProperty Relationships in Diketopyrrolopyrrole-Based Polymer Semiconductors via Side-Chain Engineering. Chem. Mater. 2015, 27, 1732−1739. Yao, J. J.; Yu, C. M.; Liu, Z. T.; Luo, H. W.; Yang, Y.; Zhang, G. X.; Zhang, D. Q. Significant Improvement of Semiconducting Performance of The Diketopyrrolopyrrole-Quaterthiophene Conjugated Polymer Through Side-Chain Engineering via Hydrogen-Honding. J. Am. Chem. Soc. 2016, 138, 173−185. Yu, H.; Kim, H. N.; Song, I.; Ha, Y. H.; Ahn, H.; Oh, J. H.; Kim, Y. H. Effect of Alkyl Chain Spacer on Charge Transport in N-type Dominant Polymer Semiconductors with a Diketopyrrolopyrrole-Thiophene-Bithiazole Acceptor-Donor-Acceptor Unit. J. Mater. Chem. C. 2017, 5, 3616−3622. Cai, Y.; Liang, P. P.; Tang, Q. Y.; Yang, X. Y.; Si, W. L.; Huang, W.; Zhang, Q.; Dong, X. C. DiketopyrrolopyrroleTriphenylamine Organic Nanoparticles as Multifunctional Reagents for Photoacoustic Imaging-Guided Photodynamic/Photothermal Synergistic Tumor Therapy. ACS Nano 2017, 11, 1054−1063. Wang, L. Y.; Zhuo, S. C.; Tang, H.; Cao, D. R. An Efficient Fluorescent Probe for Rapid Sensing of Different Concentration Ranges of Cysteine with Two-stage Ratiometric Signals. Dyes and Pigments. 2018, 157, 284−289. Hang, Y. D.; He, X. P.; Yang, L.; Hua, J. L. Probing SugarLectin Recognitions in the Near-Infrared Region Using Glyco-

40.

41.

42.

43.

44.

45.

46.

47.

48.

49.

50.

51.

Diketopyrrolopyrrole with Aggregation-Induced-Emission. Biosens. Bioelectron. 2015, 65, 420−426. Hang, Y. D.; Wang, J.; Jiang, T.; Lu, N. N.; Hua, J. L. Diketopyrrolopyrrole-Based Ratiometric/Turn-on Fluorescent Chemosensors for Citrate Detection in the Near-Infrared Region by an Aggregation-Induced Emission Mechanism. Anal. Chem. 2016, 88, 1696−1703. Zhang, X.; Hang, Y. D.; Qu, W. S.; Yan, Y. C.; Zhao, P.; Hua, J. L. Diketopyrrolopyrrole-Based Ratiometric Fluorescent Probe for The Sensitive and Selective Detection of Cysteine Over Homocysteine and Glutathione in Living Cells. RSC Adv. 2016, 6, 20014−20020. Wang, L. Y.; Chen, X. G.; Cao, D. R. A Novel Fluorescence Turn-on Probe Based on Diketopyrrolopyrrole-nitroolefin Conjugate for Highly Selective Detection of Glutathione over Cysteine and Homocysteine. Sensor. Actuat. B-Chem. 2017, 244, 531−540. Wang, L. Y.; Yang, L. L.; Huang, L. B.; Cao, D. R. Diketopyrrolopyrrole-derived Schiff Base as Colorimetric and Fluoromertic Probe for Sequential Detection of HSO4- and Fe3+ with “off-on-off” Response. Sensor. Actuat. B-Chem. 2015, 209, 536−544. Wang, L. Y.; Wu, S. M.; Tang, H.; Meier, H.; Cao, D. R. An Efficient Probe for Sensing Different Concentration Ranges of Glutathione Based on AIE-active Schiff Base Nanoaggregates with Distinct Reaction Mechanism. Sensor. Actuat. B-Chem. 2018, 273, 1085−1090. Shen, Y. M.; Zhang, X. Y.; Zhang, Y. Y.; Wu, Y. Y.; Zhang, C. X.; Chen, Y. D.; Jin, J. L.; Li, H. T. A Mitochondria-Targeted Colorimetric and Ratiometric Fluorescent Probe for Hydrogen Peroxide with A Large Emission Shift and Bio-imaging in Living Cells. Sensor. Actuat. B-Chem. 2018, 255, 42−48. Ren, M. G.; Deng, B. B.; Zhou, K.; Kong, X. Q.; Wang, J. Y.; Lin, W. Y. Single Fluorescent Probe for Dual-Imaging Viscosity and H2O2 in Mitochondria with Different Fluorescence Signals in Living Cells. Anal. Chem. 2017, 89, 552−555. Halabi, E. A.; Thiel, Z.; Trapp, N.; Pinotsi, D.; Rivera-Fuentes, P. A Photoactivatable Probe for Super-Resolution Imaging of Enzymatic Activity in Live Cells. J. Am. Chem. Soc. 2017, 139, 13200−13207. Abney, K. K.; Ramos-Hunter, S.; Romaine, I. M.; Goodwin, J. S.; Sulikowski, G. A.; Weaver, C. D. Selective Activation of N,N’-Diacyl Rhodamine Pro-fluorophores Paired with Releasing Enzyme, Porcine Liver Esterase (PLE). Chem. Eur. J. 2018, 24, 8985−8988. Kaouther, B. A.; Pieter, B.; Patrick, V.; Frank, M. R.; Antoon, D. L. A.; Willem, M. D. V.; Tjakko, A. Multiparametric Flow Cytometry and Cell Sorting for the Assessment of Viable, Injured, and Dead Bifidobacterium Cells during Bile Salt Stress. Appl. Environ. Microbiol. 2002, 68, 5209−5216. Phillips, C. B.; Iline, I. I.; Novoselov, M.; Richards, N. K.; Potential to Exploit Postmortem Enzyme Degradation for Evaluating Arthropod Viability. Appl. Entomol. Zool 2014, 49, 421−428. Wang, J.; Hang, Y. D.; Tan, H. Q.; Jiang T.; Qu, X.; Hua, J. L. Two New Colorimetric and Ratiometric Fluorescent Probes Based on Diketopyrrolopyrrole (DPP) for Detecting and Imaging of Mitochondrial SO2 Derivatives in Cancer Cells. J. Photoch Photobio A 2017, 346, 265−272.



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