Spin-Labeled Diclofenac: Synthesis and Interaction with Lipid Membranes
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis
2.2. EPR Spectra: Interaction with the POPC Membrane
2.3. Pulsed EPR: Location in the POPC Membrane
3. Materials and Methods
3.1. Chemical Analysis
3.2. Synthesis and Characterization
- {2-[(2,6-Dichlorophenyl)amino]-5-iodophenyl}acetic acid (2). KI (1.8 g, 10.8 mmol) was added in several portions to a stirred mixture of diclofenac (3.2 g, 10.8 mmol), NaCl (1.26 g, 21.5 mmol), NaIO4 (2.3 g, 10.8 mmol) and H2O (10 mL) in AcOH (60 mL) at room temperature for 24 h. After the disappearance of the iodine color, the precipitate was filtered, washed with H2O and dried in air. The crude product was purified with flash chromatography on silica gel (eluent: ethyl acetate). Yield 4.0 g (88%), white solid, mp 191–193 °C (toluene). 1H-NMR (500 MHz, DMSO-D6) δ: 7.53 (m, 3H), 7.35 (d, J = 8.4 Hz, 1H), 7.31 (s, 1H), 7.23 (m, 1H), 6.04 (d, J = 8.4 Hz, 1H), 3.68 (s, 2H) (see 1H-NMR in [43,44]). IR (film) cm−1: 3352 (NH), 3182, 3028, 2814 (OH), 1693 (C=O).
- Methyl {2-[(2,6-dichlorophenyl)amino]-5-iodophenyl}acetate (3). A mixture of acid 2 (2.0 g, 4.75 mmol) and concentrated H2SO4 (1 mL) in methanol (45 mL) was stirred at reflux for 0.3 h. Then, the reaction mixture was cooled to room temperature; the precipitate was isolated using filtration, washed with methanol and dried in air. Yield 1.96 g (95%), white solid, mp 119–120 °C (methanol). 1H-NMR (500 MHz, CDCl3) δ: 7.53 (d, J = 1.5 Hz, 1H), 7.39 (dd, J = 8.5, 1.8 Hz, 1H), 7.35 (d, J = 8.0 Hz, 2H), 7.01 (t, J = 8.0 Hz, 1H), 6.94 (br.s, 1H), 6.28 (d, J = 8.5 Hz, 1H), 3.76 (s, 3H), 3.75 (s, 2H). 13C-NMR (126 MHz, CDCl3) δ: 172.24, 142.83, 139.44, 137.19, 136.95, 129.96, 129.08, 126.29, 124.78, 119.97, 84.13, 52.73, 38.16. IR (film) cm−1: 3321 (NH), 2949 (Me), 1714 (C=O). HRMS (ESI) m/z: [M]+ Calcd for C15H12Cl2INO2 434.9284. Found 434.9285.
- Methyl {2-[(2,6-dichlorophenyl)amino]-5-[(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)ethynyl]phenyl}acetate (4). Under an argon atmosphere, a mixture of iodide 3 (500 mg, 1.15 mmol), PdCl2(PPh3)2 (20 mg, 0.03 mmol) and CuI (10 mg, 0.05 mmol) in toluene (20 mL) was stirred and heated to 55 °C. Then, Et3N (8 mL) and 4-ethynyl-2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridine (270 mg, 1.65 mmol) were added, and the reaction mixture was stirred at 75 °C for 16 h. After cooling, toluene (100 mL) was added and the mixture was filtered. The extract was washed with 6 M of aqueous ammonia (2 × 200 mL) and dried over MgSO4. The crude product was purified with column chromatography on silica gel (eluent: toluene/ethyl acetate, 4:1). Yield 470 mg (87%), white solid, mp 150–151 °C (ethyl acetate). 1H-NMR (500 MHz, CDCl3) δ: 7.34 (m, 3H, HAr), 7.19 (m, 1H), 7.11 (s, 1H), 7.01 (t, J = 8.0 Hz, 1H), 6.45 (d, J = 8.2 Hz, 1H), 6.05 (s, 1H), 3.76 (s, 2H), 3.75 (s, 3H), 2.87 (br.s, 1H), 2.07 (s, 2H), 1.25 (s, 6H), 1.20 (s, 6H). 13C-NMR (126 MHz, CDCl3) δ: 172.52, 142.71, 140.23, 137.06, 134.25, 131.34, 129.90, 129.04, 124.68, 123.59, 117.68, 116.46, 116.33, 89.61, 87.68, 52.70, 51.91, 49.71, 41.03, 38.46, 30.94, 29.75. IR (film) cm−1: 3348 (NH), 2961 (Me), 1745 (C=O). HRMS (ESI) m/z: [M]+ Calcd for C26H28Cl2N2O2 470.1522. Found 470.1521.
- [4-{[3-(Carboxymethyl)-4-((2,6-dichlorophenyl)amino)phenyl]ethynyl}-2,2,6,6-tetramethyl-3,6-dihydropyridin-1(2H)-yl]oxidanyl (1). A mixture of compound 4 (120 mg, 0.25 mmol), Na2WO4·2H2O (13 mg, 0.04 mmol), EDTA disodium salt (13 mg, 0.04 mmol) and 30% H2O2 (0.3 mL) in 1,4-dioxane (3 mL) was stirred at the ambient temperature for 14 days. The suspension was filtered, and the solvent was evaporated under a vacuum condition. Then, 2 M of NaOH (3 mL) was added, and the reaction mixture was stirred at 80 °C for 0.5 h. Next, the resulting mixture was diluted with ethanol (3 mL) and stirred at reflux for 0.25 h. After cooling, H2O (5 mL) and CH2Cl2 (15 mL) were added. At 0 °C, 1 M of HCl was added dropwise to the stirred mixture until the pH was neutral. The organic layer was separated and dried over MgSO4. The solvent was evaporated under a vacuum condition. The crude product was purified with column chromatography on silica gel (eluent: toluene/ethyl acetate, 5:1). Yield 40 mg (34%), white solid, mp 190–192 °C (toluene). 1H-NMR (500 MHz, CDCl3) δ: 7.51 (m, 4H), 7.10 (s, 1H), 6.95 (s, 1H), 6.46 (s, 1H), 3.98 (s, 2H). 13C-NMR (126 MHz, CDCl3) δ: 176.065, 140.80, 135.85, 130.19, 128.73, 128.02, 127.33, 123.77, 122.90, 119.32, 117.51, 38.66. IR (film) cm−1: 3300 (NH), 3042 (OH), 2928, 2851 (Me), 2203 (C≡C), 1695 (C=O). HRMS (ESI) m/z: [M]+ Calcd for C25H25Cl2N2O3 471.1237. Found 471.1235.
3.3. Sample Preparations for EPR Investigation
3.4. EPR Measurements
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Baranov, D.S.; Kashnik, A.S.; Atnyukova, A.N.; Dzuba, S.A. Spin-Labeled Diclofenac: Synthesis and Interaction with Lipid Membranes. Molecules 2023, 28, 5991. https://doi.org/10.3390/molecules28165991
Baranov DS, Kashnik AS, Atnyukova AN, Dzuba SA. Spin-Labeled Diclofenac: Synthesis and Interaction with Lipid Membranes. Molecules. 2023; 28(16):5991. https://doi.org/10.3390/molecules28165991
Chicago/Turabian StyleBaranov, Denis S., Anna S. Kashnik, Anastasiya N. Atnyukova, and Sergei A. Dzuba. 2023. "Spin-Labeled Diclofenac: Synthesis and Interaction with Lipid Membranes" Molecules 28, no. 16: 5991. https://doi.org/10.3390/molecules28165991