Trimethyl-Substituted Carbamate as a Versatile Self-Immolative Linker for Fluorescence Detection of Enzyme Reactions
Abstract
:1. Introduction
2. Results
2.1. Development of N-Methyl Dimethyl Methyl Carbamate Linker
2.2. Fluorescence Detection of Enzyme Reactions with the Trimethyl Carbamate Linker
3. Materials and Methods
3.1. Fluorescence Measurement and Reaction Kinetics Analysis of the Coumarin Probes
3.2. Fluorescence Detection of Esterase Activity Using Probe 17
3.3. Fluorescence Detection of Ketoreductase Activity Using Probe 16
3.4. Fluorescence Detection of Transaminase Activity Using Probe 16
3.5. Cell Culture
3.6. Evaluation of Cell Viability after Treatment of A549 Cells with Probe 18
3.7. Evaluation of Reaction of 18 and Glutathione
3.8. Fluorescence Imaging of Intracellular Esterases in Living A549 Cells
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
1R | |||||
2 | 3 | 4 | 5 | 6 | |
k (h−1) | n.d. b | 0.023 | 1.2 | 120 | n.d. c |
t1/2 (h) | n.d. b | 30 | 0.58 (35 min) | 0.0058 (21 s) | > 100 h |
1R | |||||
7 | 8 | 9 | 10 | 11 | |
k (h−1) | n.d. c | 1.3 | 36 | 0.060 | n.d. c |
t1/2 (h) | > 100 h | 0.53 (32 min) | 0.019 (1.2 min) | 11.6 | > 100 h |
1R | |||
5 | 12 | 13 | |
k (h−1) | 120 | 92 | n.d. b |
t1/2 (h) | 0.0058 (21 s) | 0.021 (75 s) | > 100 h |
1R | |||
14 | 15 | 16 | |
k (h−1) | 0.64 | 0.75 | n.d.b |
t1/2 (h) | 1.1 h | 0.92 h | > 100 h |
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Nakamura, N.; Uchinomiya, S.; Inoue, K.; Ojida, A. Trimethyl-Substituted Carbamate as a Versatile Self-Immolative Linker for Fluorescence Detection of Enzyme Reactions. Molecules 2020, 25, 2153. https://doi.org/10.3390/molecules25092153
Nakamura N, Uchinomiya S, Inoue K, Ojida A. Trimethyl-Substituted Carbamate as a Versatile Self-Immolative Linker for Fluorescence Detection of Enzyme Reactions. Molecules. 2020; 25(9):2153. https://doi.org/10.3390/molecules25092153
Chicago/Turabian StyleNakamura, Noriaki, Shohei Uchinomiya, Kazuya Inoue, and Akio Ojida. 2020. "Trimethyl-Substituted Carbamate as a Versatile Self-Immolative Linker for Fluorescence Detection of Enzyme Reactions" Molecules 25, no. 9: 2153. https://doi.org/10.3390/molecules25092153