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, Anastasia Parchina +
Hannu Myllykallio ,
,
, Prof. E. Lescrinier Medicinal Chemistry, vol.49
, Prof. H. F. Becker Laboratory of Optics and Biosciences, INSERM U 696-CNRS UMR 7645, 91128.
, Becker FacultØ des Sciences et IngØnierie, Sorbonne UniversitØ, 4 place Jussieu 75005
Jonghe Present affiliation: Laboratory of Virology and Chemotherapy, Herestraat, vol.49 ,
Vanhoutte Present affiliation: Laboratory of Chemical Biology, KU Leuven, O&N I, Herestraat 49, PO Box, vol.802 ,
, Supporting information and the ORCID identification number(s) for the author(s) of this article can be found under
,
, 6.93 (t, J = 7.2 Hz, H, CH), 7.33 (t, J = 7.55 Hz, 2 H, 2CH), 7.45 (d
, (3H)-yl)propanoyl)-N-phenylpiperazine-1-carboxamide (17 a): A mixture of 6-chloro-2H-benzo
, Cl 2 (100 mL) and washed with H 2 O (40 mL), dried over Na 2 SO 4 , concentrated and purified by flash column chromatography to afford the title compound (318 mg, 72 %). Purity (Method A): 95.86 %. 1 H NMR (300 MHz
CDCl 3 ): d = 30, vol.69 ,
,
58 (s, 2 H, O-CH 2 -CO), 4.23 (t, 2 H, 300 MHz, CDCl 3 ): d = 7.1-6.95 (m, 4 H), vol.4 ,
,
,
ppm (t, 2 H, J = 7.5 Hz, N-CH 2 -CH 2 ), vol.13 ,
,
, Oxo-3-(piperazin-1-yl)propyl)-2H-benzo
, MeOH/NH 3(aq) in a ratio of 98:2:0.3), to yield the title compound as a light-yellow oil (0.4 g, 31 %). 1 H NMR (300 MHz
,
, CDCl 3 ): d = 7.40 (s, 1 H), 7.30-6.93 (m, 8 H), 4.57 (s, 2 H, O-CH 2 -CO, Purity (Method A): 99.83 %. 1 H NMR (300 MHz
, , vol.8, p.30
, HRMS (ESI)
, Oxo-2,3-dihydro-4H-benzo, vol.27
92:8:0.3). DIPEA (42an off-white solid (28 mg, 32 %), Purity (Method A, vol.3 ,
, CDCl 3 ): d = 8.38-8.32 (m, 2 H), vol.7
, 57 (s, 2 H, O-CH 2 -CO), 4.24 (t, 2 H, J = 7.6 Hz, N-CH 2 -CH 2 -CO, vol.4
, Compound 28 was synthesized according to the procedure for the preparation of compound 27. Exact experimental and spectral data can be found in the Supporting Information. 2-(4-(3-(3-Oxo-2H-benzo[b][1,4]oxazin-4(3H)-yl)propanoyl)piperazin-1-yl)-N-(pyridin-3-yl)acetamide (29 a): To a solution of 2-(piperazin-1-yl)-N-(pyridin-3-yl)acetamide (110 mg, 0.5 mmol) in DMSO (10 mL) was added compound 20 (110 mg 0.50 mmol), HCTU (206 mg 0.5 mmol) and DIPEA (50 mL). The reaction mixture was stirred overnight at room temperature. Then, the mixture was diluted with CH 2 Cl 2 (100 mL) and washed with H 2 O (40 mL). The combined organic layers were dried over Na 2 SO 4 and concentrated in vacuo, HRMS (ESI): m/z [M + H] + calculated for C 21 H 24 N 5 O 4 410.18226, found 410.1817
19 (s, 2 H, CH 2 ), 3.56-3.58 (m, 2 H, (m, 4 H, 4CH), 7.27-7.31(m, 1 H, 1CH), 8.19-8.23 (m, 1 H, 1CH), 8.32-8.37 (m, 1 H, 1CH), 8.59 (d, J = 2.4 Hz, H, CH), 9.05 ppm, vol.3 ,
, 00 g, 33.52 mmol) and K 2 CO 3 (13.90 g, 100.57 mmol) were dissolved in DMF (80 mL) and stirred at room temperature for 15 min. Then, ethyl bromoacetate (11.20 g, 67 mmol) was added and the reaction mixture was stirred overnight at 90 8C. The solvents were evaporated in vacuo and the residue was purified by silica gel column chromatography (using a mixture of cyclohexane and EtOAc in a ratio of 7:3 as mobile phase) affording the title compound as a colorless oil (7.70 g, 98 %). 1 H NMR (300 MHz, CDCl, vol.18, issue.5
, 1 H NMR (300 MHz, DMSO): d = 13.09 (br s, 1 H), 7.10-6.95 (m
, ChemMedChem, vol.14, pp.1-19, 2019.
Weinheim 4-(2-Oxo-2-(piperazin-1-yl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (30''): 2-(3-Oxo-2,3-dihydro-4H-benzo, 2019. ,
, 300 MHz, DMSO): d = 7.07-6.98 (m, 3 H), vol.6
, m, 2 H), 3.42-3.36 (m, 2 H), 2.84-2.77 (m, 2 H), 2.73-2.66 ppm (m, 2 H
, HRMS (ESI)
, Compound 32 was synthesized according to the procedure described for the synthesis of compound 22 a, affording the title compound as a white powder (89 mg, 89 %). Purity (Method A): 98.76 %. 1 H NMR (300 MHz, CDCl 3 ): d = 7.40-7.25 (m, 3 H), 7.10-6.95 (m, 4 H)
, HRMS (ESI)
, 3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)propyl)-Nphenylpiperazine-1-carboxamide (39): A mixture of 2H-benzo
, Purity (Method A): 98.85 %. 1 H NMR (300 MHz, vol.101, pp.33-40
, CDCl 3 ): d = 24, vol.41
, 4-Diazepan-1-yl)-3-oxopropyl)-2H-benzo
, Cl 2 /MeOH/ NH 3 (aq) 92:8:0.3), to yield the title compound as a light-yellow oil (0.506 g, 46 %). 1 H NMR (300 MHz, reaction mixture was stirred overnight at room temperature. The precipitate was filtered off and washed with CH 2 Cl 2 . The filtrate was evaporated and the residue was purified by silica gel flash chromatography (the mobile phase being CH 2, vol.6, pp.98-104
, This compound was synthesized from compound 40, according to the procedure for the synthesis of compound 22 a. The crude residue was purified by silica gel flash chromatography (using EtOAc as mobile phase), to yield the title compound as a white powder (93 % yield), Purity (Method A, issue.3
, (m, 2 H), 2.75-2.65 (m, 1.98 ppm
, mmol) was dissolved in THF (10 mL) and a solution of phenylisocyanate (60 mg 0.57 mmol) in THF (5 mL) was slowly added at room temperature. A light-yellow solid was formed upon reaction completion. Then, trifluoroacetic acid (10 mL) was added dropwise while stirring at 0 8C. When TLC showed completion of the deprotection, the mixture was diluted with H 2 O (50 mL). Then, the mixture was washed with CH 2 Cl 2 (30 mL) and the aqueous phase was adjusted to pH 9 by the addition of 1 n NaOH. The mixture was extracted with CH 2 Cl 2 (50 mL). The combined organic layers were dried over Na 2 SO 4 . The solvents were evaporated in vacuo, affording crude 44 a. This solid was used in the next reaction without further purification. The solid was re-dissolved in DMSO (10 mL), Purity (Method A
ppm (s, H, NH); 13 C NMR (75 MHz ,
, ]oxazin-4-yl)propanoyl)-N-phenylpiperazine-1-carboxamide (48): A mixture of 2H-pyrido, vol.2
, ChemMedChem, vol.14, pp.1-19, 2019.
, Weinheim (158 mg, 1.0 mmol), DMF (3 mL, p.3, 2019.
, CDCl 3 ): d = 2.76-2.81 (m, 2 H
, , vol.7
ppm (dd, J = 4.9, 1.5 Hz, 1 H, CH Ar ), vol.13 ,
, Oxoquinoxalin-1(2H)-yl)propanoyl)-N-phenylpiperazine-1-carboxamide (62): A mixture of tert-butyl 3-oxo-3,4-dihydroquinoxaline-1(2H)-carboxylate 60 (248 mg, 1 mmol, vol.2
, DCC (206 mg, 1 mmol) purified by flash column chromatography (EtOAc/MeOH 10:1) to yield the title compound (102 mg, 25 %). Purity (Method A): 98.86 %. 1 H NMR (300 MHz, The acid was dissolved in DMF (10 mL), and HOBt (153 mg, 1 mmol)
HRMS (ESI): m/z [M + H] + calculated for C 22 H 24 N 5 O 3 406.18737, found 406.1864. 4-(3-(2-Oxobenzo[d]oxazol-3(2H)-yl)propanoyl)-N-phenylpiperazine-1-carboxamide (64): 3-(2-Oxobenzo[d]oxazol-3(2H)-yl)propanoic acid 63 (104 mg, 0.5 mmol) was dissolved in DMF (5 mL) and HOBt (76 mg, 0.5 mmol) and DCC (103 mg, 0.5 mmol) were added at 0 8C. After stirring for 2 h at 0 8C, N-phenylpiperazine-1-carboxamide 16 (102 mg, 0.5 mmol) was added. The mixture was stirred overnight at room temperature. The resulting mixture was diluted with CH 2 Cl 2 (20 mL) and washed with H 2 O (2 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and concentrated in vacuo. The crude residue was purified by silica gel flash column chromatography, p.99 ,
, 4 mmol) and ethyl 3-bromopropionate (380 mg, 2.4 mmol) in DMF (3 mL) was stirred at 80 8C for 48 h. When the reaction was finished, the mixture was diluted with H 2 O (15 mL) and extracted with CH 2 Cl 2 (3 10 mL). The organic phases were combined and washed with H 2 O (15 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and evaporated in vacuo. The crude residue was dissolved in THF (5 mL) and a solution of LiOH (42 mg, 1 mmol) in H 2 O (5 mL) was added. The mixture was stirred for 18 h at 60 8C. Then, the solution was adjusted to pH 3 with 2 n HCl and the compound was extracted with EtOAc (3 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After solvent evaporation, the pure acid compound was obtained. The acid was re-dissolved in DMF (5 mL), and HOBt (76 mg, 0.5 mmol) and DCC (103 mg, 0.5 mmol) were added at 0 8C. After stirring for 2 h, N-phenylpiperazine-1-carboxamide 16 (102 mg, 0.5 mmol) was added. The mixture was stirred overnight at room temperature. The resulting mixture was diluted with CH 2 Cl 2 (20 mL) and washed with H 2 O (2 20 mL). The organic phase was dried over Na 2 SO 4 and concentrated in vacuo. The crude residue was purified by silica gel flash column chromatography, vol.14, pp.1-19, 2019.
, Weinheim Biological assays, 2019.
, The synthesized compounds were screened using a NADPH oxidation assay for M. tuberculosis ThyX activity in 96-well plates. [27] To determine IC 50 values of compounds 22 g, 27, 28, 29 d, 45 c, 48 and 52, the reaction mixture (100 mL) contained 50 mm HEPES pH7.5, 1 mm MgCl 2 , 12 mm FAD, 0.5 % glycerol, 0.05 % Triton X-100, 0.5 mg mL À1 BSA, Mycobacterial ThyX NADPH oxidase assay: M. tuberculosis ThyX was produced and purified as described
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