In a NMR tube with a J Young valve were added a solution of 2-azidoaniline (16 .... Radiochemical purity (RCP) was calculated as the ratio of the area under the ...
Supporting Information
Late-Stage Isotopic Carbon Labeling of Pharmaceutically Relevant Cyclic Ureas Directly from CO2 Antonio Del Vecchio, Fabien Caill, Arnaud Chevalier, Olivier Loreau, Kaisa Horkka, Christer Halldin, Magnus Schou, Nathalie Camus, Pascal Kessler, Bertrand Kuhnast, Frdric Taran, and Davide Audisio* anie_201804838_sm_miscellaneous_information.pdf
Supporting Information Table of Contents I. General Information........................................................................................................................................................... S2 II. Synthetic protocols ........................................................................................................................................................... S3 II. 2. Preparation of aliphatic and aromatic secondary amines through reductive amination ......................................... S8 II. 3. Preparation of new analogues of aminoazides ....................................................................................................... S13 II. 4. Synthesis of drug precursors. .................................................................................................................................. S17 II. 5. Preparation of [13C]benzo[d]imidazol-2(3H)-one and [13C]tetrahydropyrimidin-2(1H)-one derivatives................ S25 II. A. Optimization of the Staudinger-Aza-Wittig reaction (SAW) ............................................................................ S25 II. 6. Synthesis of [13C]Drug derivatives ........................................................................................................................... S37 II. 7. Experimental procedure for Carbon-14 radiolabeling ............................................................................................ S40 II. 8. Experimental procedure for Carbon-11 radiolabeling. ........................................................................................... S44 II. 9. Experimental procedure for Carbon-11 radiolabeling (Karolinska Institutet, Stockholm, Sweden) ...................... S52 III. 1H and 13C NMR Data...................................................................................................................................................... S56 IV. Radio-HPLC Analysis for 11C-Labeled Compounds V. Radio-TLC Analysis for 14C-Labeled Compounds
S104 S138
S1
I. General Information Organic solvents (Aldrich) were used without further purification. Purifications of reactions products were carried out by flash chromatography using Merck silica gel (40-63 µm). FT-ATR-IR spectra were recorded on a Perkin-Elmer UAR Two Spectrum spectrometer and are reported as wavelength numbers (cm-1). 1H NMR (400 MHz),13C NMR (100 MHz) were measured on a Brucker Avance 400 MHz spectrometer. Chemical shifts are reported in parts per million (ppm, δ) downfield from residual solvents peaks and coupling constants are reported as Hertz (Hz). Splitting patterns are designated as singlet (s), doublet (d), triplet (t). Splitting patterns that could not be interpreted or easily visualized are designated as multiplet (m). Electrospray mass spectra were obtained using an ESI-Quadripole autopurify, Waters (pump 2545, mass: ZQ2000) mass Spectrometer. Unless otherwise noted, all other commercially available reagents and solvents were used without further purification. Carbonylations for 13C experiments were performed using carbon-13 CO2 (99.2% enrichment) provided by Isotec.inc
S2
II. Synthetic protocols II. 1. Preparation of the aromatic azido derivatives via direct N3-insertion General procedures for the synthesis of azido-derivatives according to the procedures of Jiao1 (A) and Hao2 (B)
Procedure A1 To a stirred solution of aniline derivatives in CH3CN is added CuBr (0.1 equiv.) followed by TMSN3 (2.0 equiv.) and TBHP 6 M in decane (2.0 equiv.). The mixtures were stirred under argon at RT for a time varying from 2 hours to overnight then were filtered over Celite. The crude was purified by Combiflash (40 g column) affording the corresponding azides. Procedure B2 To a stirred solution of 1-azido-1,2-benziodoxol-3-(1H)-one (1.00 mmol) in THF (5.0 mL) were sequentially added Cu(OAc)2 (0.22 mmol) and a solution of the corresponding aniline (2.00 mmol) in THF (1.50 mL). The mixtures were stirred overnight at RT then were diluted with EtOAc and washed with saturated aqueous NaHCO3. The organic phases were dried over MgSO4, filtered, concentered under reduced pressure and purified by Flash Chromatography affording the corresponding azides.
2-azidoaniline (1a) C6H6N4 MW: 134.14 g.mol-1 Yield: 40% Brown solid 2-azidoaniline 1a was prepared accordingly to the procedure A1 using aniline (0.98 mL, 10.8 mmol), CuBr (0.154 mg, 1.08 mmol), TMSN3 (3.01 mL, 21.6 mmol) and TBHP 6 M in decane (3.6 mL) in CH3CN (20 mL). The mixture was stirred under argon at 30 °C for 2 hours and 30 min then was filtered over Celite. The crude product was purified by Combiflash (from cHex to 10% EtOAc) affording 2-azidoaniline 1a as a brown solid (580 mg, 40%). The spectral data (1H-NMR) was consistent with reported one1. 1
H NMR (400 MHz, CDCl3): 7.06 (dd, 1H), 6.99 (td, 1H), 6.82 (td, 1H), 6.72 (dd, 1H). IR (cm-1) 3385, 3186, 2119, 1588, 1495, 1289, 1258, 1153, 740, 702, 648, 552. LCMS (ESI) m/z C6H6N4 [(M-N2)+H]+ 106.9.
1
J. Am. Chem. Soc., 2012, 134 (46), 18924–18927
2
Chem. Commun., 2014, 50, 5733-5736 S3
2-Azido-4-Chloroaniline (S1) C6H5ClN4 MW: 168.58 g.mol-1 Yield: 38% Brown solid 2-azido-4-chloroaniline S1 was prepared accordingly to the procedure A1 using 4-chloroaniline (0.50 g, 3.9 mmol), CuBr (0.39 mmol, 56.0 mg), TMSN3 (7.80 mmol, 1.04 mL) and TBHP 6 M in decane (1.0 mL) in CH3CN (8.0 mL). The mixture was stirred under argon at 30 °C for 2 hours and 30 minutes then was filtered over Celite. The crude product was purified by Combiflash (from cHex to 10% EtOAc) affording the 2-azido-chloroanaline S1 as a brown solid (250 mg, 38%). The spectral data (1H-NMR) was consistent with reported one1. 1
H NMR (400 MHz, CDCl3) δ 7.11 (d, J = 8.2 Hz, 2H), 7.78 (s, 1H), 7.77–7.67 (m, 3H). IR (cm-1) 3424, 3316, 2100, 1584, 1489, 1274, 1253, 1152, 886, 846, 803, 694, 646, 559, 520. LCMS (ESI) m/z C6H5ClN2 [M+H]+ 169.2. 2-azido-4-fluoroaniline (S2) C6H5FN4 MW: 152.13 g.mol-1 Yield: 53% Brown solid 2-azido-4-fluoroaniline S2 was prepared according to the procedure B2 using a solution of 2,4-dimethylaniline (190 µL, 2.0 mmol) in THF (1.50 mL), Cu(OAc)2 (38.3 mg, 0.22 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (293 mg, 1.0 mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 9:1) afforded the 2-azido-4-fluoroaniline S2 as a brown solid (80 mg, 53%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, DMSO-d6) δ 6.9 (dd, J = 9.3, 2.6 Hz, 1H), 6.72 (m, 2H), 4.11 (br, s, 2H). IR (cm-1) 3491.6, 3309.1, 3192.4, 2108.2, 1600.1, 1503.8, 1432.2, 1310.7, 1274.6, 1253, 1200, 950, 840, 797, 650, 575. LCMS (ESI) m/z C6H5FN4 [(M-N2)+H]+ 127. 2-azido-4-iodoaniline (S3) C6H5IN4 MW: 260.04 g.mol-1 Yield: 25% Brown oil 2-azido-4-iodoaniline S3 was prepared according to the procedure B2 using a solution of 4-iodoaniline (448 mg, 2.0 mmol) in THF (1.50 mL), Cu(OAc)2 (38.3 mg, 0.22 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (289 mg, 1.0 mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 9:1) afforded the 2-azido-4-iodoaniline S3 as a brown oil (66 mg, 25%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, CDCl3) δ 7.25 (t, J= 14.1 Hz, 2H), 6.73 (d, J = 8.3 Hz, 1H), 4.12 (br, s, 2H). IR (cm-1) 3469, 3376, 2112, 1612, 1577, 1491, 1406, 1326, 1298, 1262, 1156, 862, 806, 653, 554. LCMS (ESI) m/z C6H5IN4 [M+H]+ 262.2. S4
2-azido-4-methoxyaniline (S4)
C7H8N4O MW: 164.17 g.mol-1 Yield: 31% Brown solid 2-azido-4-methoxyaniline S4 was prepared according to the procedure B2 using a solution of 4-methoxylaniline (496 mg, 4.0 mmol) in THF (3.0 mL), Cu(OAc)2 (75.1 mg, 0.4 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (581 mg, 2.0 mmol) in THF (10.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 8:2) afforded the 2-azido-4methoxyaniline S4 as a brown oil (100 mg, 31%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, CDCl3) δ 6.67 (d, J = 8.6 Hz, 1H), 6.62 (d, J = 2.6 Hz, 1H), 6.56 (dd, J = 8.6, 2.6 Hz, 1H), 3.77 (s, 3H). IR (cm-1) 3435, 3359, 2110, 1591, 1509, 1431, 1299, 1284, 1258, 1232, 1117, 1041, 825, 655. LCMS (ESI) m/z C7H8N4O [M+H]+ 165. tert-butyl (4-amino-3-azidophenyl)carbamate (S5) C11H15N5O2 MW: 249.27 g.mol-1 Yield: 18% Red Solid Tert-butyl (4-amino-3-azidophenyl)carbamate S5 was prepared according to the procedure B2 using a solution of tertbutyl (4-aminophenyl)carbamate (410 mg, 2.0 mmol) in THF (3.0 mL), Cu(OAc)2 (83 mg, 0.46 mmol) and 1-azido-1,2benziodoxol-3-(1H)-one (581 mg, 2.0 mmol) in THF (10.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 7:3) afforded tert-butyl (4-amino-3-azidophenyl)carbamate S5 as a red solid (87 mg, 18%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, DMSO-d6) δ 9.07 (br s, 1H), 7.30 (br s, 1H), 6.90 (br d, J = 7.1 Hz, 1H), 6.58 (d, J = 8.6 Hz, 1H), 4.69 (br s, 2H), 1.45 (s, 9H). IR (cm-1) 3335, 2977, 2113, 1697, 1602, 1518, 1367, 1293, 1244, 1161, 1054. LCMS (ESI) m/z C11H15N5O2 [M+H]+ 250. Methyl 4-amino-3-azidobenzoate (S6) C8H8N4O2 MW: 192.18 g.mol-1 Yield: 16% Brown solid Methyl-4-amino-3-azidobenzoate S6 was prepared according to the procedure B2 using a solution of methyl-4aminobenzoate (306 mg, 2.0 mmol) in THF (1.50 mL), Cu(OAc)2 (37.3 mg, 0.2 mmol) and 1-azido-1,2-benziodoxol-3-(1H)one (293 mg, 1.0 mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 8:2) afforded the Methyl 4-amino-3-azidobenzoate S6 as a brown solid (31 mg, 16%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400MHz, CDCl3) δ 7.73 (d, J = 1.7 Hz, 1H), 7.67 (dd, J = 8.3, 1.7 Hz, 1H), 6.68 (d, J = 8.3 Hz, 1H), 3.88 (s, 3H). S5
IR (cm-1) 3476, 3369, 2116, 1701, 1619, 1518, 1439, 1327, 1296, 1256, 1193, 1137, 1099, 991, 763, 657. LCMS (ESI) m/z C8H8N4O2 [(M-N2)+H]+ 165. 2-azido-4-(trifluoromethyl)aniline (S7) C7H5F3N4 MW: 202.14 g.mol-1 Yield: 8% Brown solid 2 2-azido-4-(trifluoromethyl)aniline S7 was prepared according to the procedure B using a solution of naphtalen-1-amine (287 mg, 2.0 mmol) in THF (1.50), Cu(OAc)2 (39.6 mg, 0.22 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (293 mg, 1.0 mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 9:1) afforded the 2-azido-4(trifluoromethyl)aniline S7 as a brown solid (17 mg, 8%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, CD3CN) δ 7.3 (d, J = 1.8 Hz, 1H), 7.23 (dd, J = 8.4 Hz, 2.0 Hz, 1H), 6.51 (d, J = 8.4 Hz, 1H), 4.29 (br s, 2H). IR (cm-1) 3490, 3396, 2116, 1619, 1526, 1432, 1326, 1275, 1151, 1108, 1076, 900, 873, 820, 746, 640. LCMS (ESI) m/z C7H5F3N4 [(M-N2)+H]+ 175. 2-azidonaphthalen-1-amine (S8) C10H8N4 MW: 184.07 g.mol-1 Yield: 16% Brown solid 2-azidonaphthalen-1-amine S8 was prepared according to the procedure B2 using a solution of naphtalen-1-amine (287 mg, 2.0 mmol) in THF (1.50 mL), Cu(OAc)2 (37.3 mg, 0.2 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (293 mg, 1.0 mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 8:2) afforded the 2azidonaphthalen-1-amine S8 as a brown solid (29 mg, 16%). The spectral data (1H-NMR) was consistent with reported one2. 1
H NMR (400 MHz, CDCl3) δ 7.76 (dd, J = 13.2, 8.3 Hz, 2H), 7.45 (m, 2H), 7.37 (d, J = 8.7 Hz, 1H), 7.26 (d, J = 8.8 Hz, 1H), 4.30 (br s, 2H). IR (cm-1) 3435, 3350, 2128, 2113, 1623, 1406, 1384, 1300, 799, 788, 745, 629. LCMS (ESI) m/z C10H8N4 [(M-N2)+H]+ 157. 2-azido-4,6-dimethylaniline (S9) C8H10N4 MW: 162.20 g.mol-1 Yield: 52% Brown solid 2-azido-4,6-dimethylaniline S9 was prepared according to the procedure B1 using a solution of 2,4-dimethylaniline (250 µL, 2.0 mmol) in THF (1.50 mL), Cu(OAc)2 (38.3 mg, 0.22 mmol) and 1-azido-1,2-benziodoxol-3-(1H)-one (293 mg, 1.0 S6
mmol) in THF (5.0 mL). Purification by Flash Chromatography on SiO2 gel (Hept/EtOAc 9:1) afforded the 2-azido-4,6dimethylaniline S9 as a brown solid (85 mg, 52%). The spectral data (1H-NMR) was consistent with reported one1. 1
H NMR (400 MHz, CDCl3) δ 6.74 (s, 1H), 6.69 (s, 1H), 3.63 (br. s, 2H), 2.25 (s, 3H), 2.13 (s, 3H). IR (cm-1) 3468, 3378, 2105, 1627, 1597, 1498, 1319, 1269, 1243, 839. LCMS (ESI) m/z C8H10N4 [M+H]+ 163. 3-azido-[1-1’-biphenyl]-2-amine (S10) C12H10N4 MW: 210.24 g.mol-1 Yield: 17% Brown solid
3-azido-[1-1’-biphenyl]-2-amine S10 was prepared accordingly to the procedure A1 using 2-aminobiphenil (0.50 g, 3.00 mmol), CuBr (0.30 mmol, 42.3 mg), TMSN3 (6.00 mmol, 0.78 mL) and TBHP 6 M in decane (1.0 mL) in CH3CN (8.0 mL). The mixture was stirred overnight under argon at 30 °C then was filtered over Celite. The crude product was purified by Combiflash (from cHex to 10% EtOAc) affording the 3-azido-[1-1’-biphenyl]-2-amine S10 as a brown solid (108 mg, 17%). The spectral data (1H-NMR) was consistent with reported one1. 1
H NMR (400 MHz, DMSO-d6) δ 7.46 (m, 2H), 7.42 – 7.34 (m, 3H), 7.10 (dd, J = 7.8, 1.1 Hz, 1H), 6.88 (dd, J = 7.6, 1.1 Hz, 1H), 6.77 (t, J = 7.7 Hz, 1H), 4.53 (br. s, 2H). IR (cm-1) 3448, 3363, 2101, 1589, 1465, 1434, 1301, 1258, 1235, 1071, 758, 732, 701, 641, 586. LCMS (ESI) m/z C12H10N4 [(M-N2)+H]+ 183.3.
S7
II. 2. Preparation of aliphatic and aromatic secondary amines through reductive amination 2-azido-N-benzylaniline (S11) C13H12N4 MW: 224.27 g.mol-1 Yield: 78% Brown oil
To a stirred solution of benzaldehyde (0.76 mmol, 75.7 µL) in 6.5 mL of MeOH at 0 °C, were sequentially added AcOH (2.98 mmol, 170 µL), AcONa (1.50 mmol, 122 mg) and 2-azidoaniline (0.76 mmol, 100 mg). After 10 minute NaBH3CN (0.76 mmol, 49.0 mg) was gradually added keeping the temperature at 0 °C, then the reaction was stirred at RT for 3 hours. The mixture was diluted with CH2Cl2 then vigorously stirred in presence of 2N NaOH. The organic phase was extracted from water, dried over anhydrous MgSO4 and concentrated under vacuum. Purification by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1) afforded the 2-azido-N-benzylaniline S11 as a dark brown oil (130 mg, 78%). 1
H NMR (400 MHz, CDCl3) δ 7.38 (m, 4H), 7.35 – 7.29 (m, 1H), 7.07 (dd, J = 7.8, 1.4 Hz, 1H), 7.02 (ddd, J = 8.0, 7.5, 1.4 Hz, 1H), 6.77 (td, J = 7.6, 1.3 Hz, 1H), 6.62 (dd, J = 8.1, 1.2 Hz, 1H), 4.51 (br s, 1H), 4.37 (s, 2H). 13 C NMR (100 MHz, CDCl3) δ 139.8, 139.1, 128.8, 127.4, 127.4, 125.9, 124.7, 117.9, 117.5, 111.2, 47.9. IR (cm-1) 3417, 3063, 3029, 2119, 1600, 1516, 1451, 1438, 1291, 1148, 735, 697. LCMS (ESI) m/z C13H12N4 [M+H]+ 225.5, 197.5. 2-azido-N-(naphthalen-2-ylmethyl)aniline (S12) C17H14N4 MW: 274.33 g.mol-1 Yield: 60% Brown oil
To a stirred solution of 2-Naphthaldehyde (0.56 mmol, 87.0 mg) in 5.0 mL of MeOH at 0 °C, were sequentially added AcOH (2.24 mmol, 122 µL), AcONa (1.12 mmol, 92.0 mg) and 2-azidoaniline (0.56 mmol, 75.0 mg). After 30 minutes, NaBH3CN (0.56 mmol, 37.0 mg) was gradually added keeping the temperature at 0 °C then the reaction was stirred overnight at RT. The mixture was diluted with CH2Cl2 then vigorously stirred in presence of 2N NaOH. The organic phase was extracted from water, dried over anhydrous MgSO4 and concentrated under vacuum. Purification by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1) afforded the 2-azido-N-(naphthalen-2-ylmethyl)aniline S12 as a brown oil (90 mg, 60%). 1
H NMR (400 MHz, MeOD) δ 7.81 – 7.74 (m, 4H), 7.45 (dd, J = 8.4, 1.5 Hz, 1H), 7.43 – 7.38 (m, 2H), 7.03 (dd, J = 7.8, 1.4 Hz, 1H), 6.91 – 6.84 (m, 1H), 6.66 (td, J = 7.6, 1.3 Hz, 1H), 6.55 (dd, J = 8.1, 1.3 Hz, 1H), 4.49 (s, 2H). 13 C NMR (100 MHz, MeOD) δ 141.2, 138.6, 135.0, 134.2, 129.2, 128.7, 128.6, 127.1, 126.8, 126.5, 126.4 (2C) 125.9, 118.8, 118.2, 112.6, 48.3. IR (cm-1) 3406, 2113, 2080, 1596, 1500, 1471, 1429, 1321, 1282, 1147, 819, 753, 742, 733, 486, 475, 469. LCMS (ESI) m/z C17H14N4 [(M-N2)+H]+ 248.5. S8
2-azido-N-(benzo[d][1,3]dioxol-5-ylmethyl)aniline (S13) C14H12N4O2 MW: 268.27 g.mol-1 Yield: 53% Dark yellow oil
To a stirred solution of 2,3-(Methylenedioxy)benzaldehyde (0.56 mmol, 87.0 mg) in 5.0 mL of MeOH at 0 °C, were sequentially added AcOH (2.24 mmol, 122 µL), AcONa (1.12 mmol, 92.0 mg) and 2-azidoaniline (0.56 mmol, 75.0 mg). After 30 minutes, NaBH3CN (0.56 mmol, 37.0 mg) was gradually added keeping the temperature at 0 °C then the reaction was stirred overnight at RT. The mixture was diluted with CH2Cl2 then vigorously stirred in presence of 2N NaOH. The organic phase was extracted from water, dried over anhydrous MgSO4 and concentrated under vacuum. Purification by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1) afforded the 2-azido-N-(benzo [d][1,3]dioxol5-ylmethyl)aniline 1n as a dark yellow oil (80 mg, 53%). 1
H NMR (400 MHz, CDCl3) δ 7.05-7.00 (m, 2H), 6.85-6.73 (m, 4H), 6.67-6.65 (m, 1H), 6.00 (s, 2H), 4.5 (s, 1H), 4.33 (s, 2H).
13
C NMR (100 MHz, CDCl3) δ 147.3, 145.3, 139.5, 125.8, 124.8, 121.7, 121.2, 120.4, 117.8, 117.5, 111.2, 107.74, 101, 42.1
IR (cm-1) 3421, 2110, 1601, 1517, 1427, 1457, 1268, 1249, 1048, 924, 763, 730, 722. LCMS (ESI) m/z C14H12N4O2 [M+H]+ 269.3
3-azidopropan-1-amine (S14) C3H8N4 MW: 100.12 g.mol-1 Yield: 53% Colorless oil To a stirred solution of 3-Bromopropypamine hydrobromide (3.30 g, 15.1 mmol) in 10.0 mL of H2O at 4 °C was added an aqueous solution of NaN3 (3.30 g, 50.0 mol). The reaction was stirred to reflux for 16 hours then was cooled to 0 °C and diluted with Et2O (10.0 mL). KOH pellets (4.0 g) were slowly added, the organic layer was separated and the aqueous phase was extracted with Et2O (2 x 30 mL). The combined organics phases were dried over anhydrous MgSO4, filtered and concentrated to give 1 g of 3-azidopropan-1-amine S14 as a colorless oil which can be used for the next step without any further purification. The spectral data (1H-NMR) was consistent with reported one3. 1
H NMR (400 MHz, CDCl3) δ 3.4-3.37 (m, 2H), 2.83-2.8 (m, 2H), 1.77-1.71 (m, 2H), 1.45 (s, 2H).
3
Chem. Commun. 2006, 19, 2012-2014. S9
3-azido-N-benzylpropan-1-amine (S15) C10H14N4 MW: 190.18 g.mol-1 Yield: 70% Colorless oil A solution of 3-azidopropan-1-amine (0.10 mmol, 100.0 mg) and Benzaldehyde (0.10 mmol, 100.0 μL) in MeOH (0.3 mL) was stirred for 1 hour at rt then NaBH4 (1.50 mmol, 57.0 mg) was added to the reaction mixture in small portions at 0 °C. The solution was stirred for other 2 hours at and then was diluted in diethyl ether. A 2M aqueous solution of NaOH was added dropwise then organic layer was extracted, dried over anhydrous MgSO4, filtered and concentrated to afford the crude product. Purification by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:1) afforded the 3-azido-Nbenzylpropan-1-amine S15 (13.3 mg, 70%). The spectral analysis were conformed with the reported ones4 1
H NMR (400 MHz, CDCl3) δ 7.38 – 7.30 (m, 4H), 7.27 (m, 1H), 3.79 (s, 2H), 3.39 (t, J = 6.7 Hz, 2H), 2.73 (t, J = 6.9 Hz, 2), 1.79 (p, J = 6.8 Hz, 2H), 1.37 – 1.23 (br. s, 1H).
Methyl 4-amino-3-azidobenzoate (S16) C8H12N4S MW: 196.27 g.mol-1 Yield: 66% Yellow oil A solution of 3-azidopropan-1-amine (100 mg, 1.0 mmol) and thiophene-2-carboxaldehyde (77.0 µL, 0.83 mmol) in MeOH (0.50 mL) was stirred for 2h at rt. NaBH4 (57.0 mg, 1.50 mmol) was added to the reaction mixture in small portions at 4 °C. The solution was stirred for 2 hours at RT and then diluted in Et2O. A 2M aqueous solution of NaOH was slowly added then the organic layer was washed with water and dried with MgSO4 anhydrous, filtered and concentrated. Purification by Flash Chromatography on SiO2 gel (cHex/EtOAc 1:1) afforded the methyl 4-amino-3-azidobenzoate S16 as a yellow oil (130 mg, 66%); 1
H NMR (400 MHz, CD3CN) δ 7.29-7.28 (d, J = 8.0 Hz, 1H), 6.98-6.96 (m, 2H), 3.95 (s, 2H), 3.41-3.38 (m, 2H), 2.69-2.66 (m, 2H), 1.77-1.7 (m, 2H). 13 C NMR (100 MHz, CD3CN) δ 143.8, 126.6, 125.0, 124.5, 49.5, 48.3, 46, 29.2. IR (cm-1) 2927, 2822; 2090, 1454, 1256, 696. LCMS (ESI) m/z C8H12N4S [M+H]+197.1.
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Angew. Chem., Int. Ed., 50, 10834-10838. S10
3-azido-N-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)propan-1-amine (S17) C12H15N5S2 MW: 293.41 g.mol-1 Yield: 70% Yellow oil
A solution of 3-azidopropan-1-amine (0.80 mmol, 80.0 mg) and 5-(2-Methyl-1,3-thiazol-4-yl)thiophene-2carboxaldehyde (0.66 mmol, 139.0 mg) in MeOH (0.30 mL) was stirred for 1 hour at RT then NaBH4 (1.20 mmol, 45.0 mg) was added to the reaction mixture in small portions at 0 °C. The solution was stirred for other 2 hours at RT and then was diluted in ET2O. A 2M aqueous solution of NaOH was added dropwise then organic layer was extracted, dried over anhydrous MgSO4, filtered and concentrated to afford the crude product. Purification by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:1) afforded the 3-azido-N-benzylpropan-1-amine S17 as a yellow oil (136 mg, 70 %). 1
H NMR (400 MHz, MeOD) δ 7.37 (d, J = 2.4 Hz, 1H), 7.29 (d, J = 3.6 Hz, 1H), 6.92 (d, J = 3.6 Hz, 1H), 3.92 (s, 2H), 3.35 (t, J = 6.7 Hz, 2H), 2.69 – 2.65 (m, 5H), 1.80 – 1.71 (m, 2H). 13 C NMR (100 MHz, MeOD) δ 168.3, 150.6, 143.7, 138.6, 127.8, 125.0, 112.6, 50.6, 48.7, 46.8, 29.7, 18.8 IR (cm-1) 2923, 2821, 2090, 1439, 1258, 1164, 802, 731, 677. LCMS (ESI) m/z C12H15N5S2 [M+H]+ 294.3.
2-(azidomethyl)-1H-benzo[d]imidazole (S18) C8H7N5 MW: 173.18 g.mol-1 Yield: 62% White solid To a stirring solution of 1H-Benzimidazole-2-methanol (148 mg, 1.0 mmol) in toluene (5 mL), diphenylphosphoryl azide (260 µL, 1.2 mmol) and DBU (200 µL, 1.3 mmol) were added. The reaction was stirred overnight at room temperature, diluted with water (10 mL) and extracted with AcOEt (3 × 10 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous sodium sulphate and the solvent evaporated. The residue was purified by flash column chromatography (20% AcOEt: n-hexane) to afford 2-(azidomethyl)-1H-benzo[d]imidazole S18 (108 mg, 62%). Analytical data were consistent with the reported literature.5 1
H NMR (400 MHz, CDCl3) δ 710.49 (br.s, 1H), 7.63 (dd, J = 6.1 Hz, 3.2 Hz, 2H), 7.28 (dd, J = 6.1 Hz, 3.2 Hz, 2H), 4.74 (s, 2H). IR (cm-1) 2100, 1425, 1312, 1272, 1224, 1185, 1025, 740. LCMS (ESI) m/z C8H7N5 [M+H]+ 174.2.
5
Org. Biomol. Chem., 2015,13, 11315-11330 S11
1-(5-((3-azidopropyl)amino)hexyl)-3,7-dimethyl-1H-purine-2,6(3H,7H)-dione (S19) C16H26N8O2 MW: 362.43 g.mol-1 Yield: 60% Colorless oil
A solution of 3-azidopropan-1-amine S19 (0.60 mmol, 60.0 mg), Pentoxyfilline (0.60 mmol, 172.0 mg) and AcOH (0.60 mmol, 34.0 μL) in MeOH (0.30 mL) was stirred for 1 hour at RT. After colling down to 0 °C, NaBH(OAc)3 (1.20 mmol, 45.0 mg) was added to the reaction mixture in small portions. The resulting solution was stirred for other 2 hours at RT and then was diluted in Et2O. A 2M aqueous solution of NaOH was added dropwise then organic layer was extracted, dried over anhydrous MgSO4, filtered and concentrated to afford the crude product. Purification by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:1) afforded the 1-(5-((3-azidopropyl)amino)hexyl)-3,7-dimethyl-1H-purine-2,6(3H,7H)dione S19 as a colorless oil (134 mg, 60%). 1
H NMR (400 MHz, CDCl3) δ 7.47 (s, 1H), 3.97-3.94 (m, 5H), 3.52 (s, 3H), 3.35-3.31 (t, J = 16.0 Hz, 2H), 2.72-2.58 (m, 3H), 2.20 (s, 1H), 1.76-1.70 (m, 2H), 1.65-1.57 (m, 2H), 1.51-1.45 (m, 1H), 1.39-1.29 (m, 3H), 1.02-1.01 (d, J = 4.0 Hz, 3H). 13 C NMR (100 MHz, CDCl3) δ 155.2, 151.4, 148.7, 141.4, 107.6, 53.1, 49.6, 44.1, 41.2, 36.3, 33.5, 29.7, 26.4, 28.1, 23.3, 20.0. IR (cm-1) 3406, 3056, 2851, 2114, 1597, 1501, 1471, 1430, 1283, 1241, 1146, 1102, 818, 732, 662, 473. LCMS (ESI) m/z C16H26N8O2 [M+H]+ 363.1.
S12
II. 3. Preparation of new analogues of aminoazides
tert-butyl (R)-(2-hydroxy-1-phenylethyl)carbamate (S20) C13H19NO3 MW: 237.30 g.mol-1 Yield: 91% White solid To a stirred solution of (R)-2-Amino-2-phenylethanol (3.64 mmol, 500.0 mg) in 8.0 mL of dry THF at 0 °C, were sequentially added NEt3 (4.37 mmol, 609.0 µL) and Boc2O (3.65 mmol, 796.0 mg). The resulting reacting mixture was stirred under argon at 0 °C for 3 hours. After the reaction was completed, the mixture was diluted in 50.0 mL of EtOAc and washed twice with 30.0 mL of a NH4Cl saturated solution. The organic phase was dried over anhydrous MgSO4 and concentrated under vacuum. The tert-butyl (R)-(2-hydroxy-1-phenylethyl)carbamate S20 was obtained as a white solid (781 mg, 91%). 1
H NMR (400 MHz, CDCl3) δ 7.36 (m, 2H), 7.28 (m, 3H), 5.29 (s, 1H); 4.77 (s, 1H), 3.82 (s, 2H), 2.46 (brs, 1H), 1.43 (s, 9H). C NMR (100 MHz, CDCl3) δ 156.3, 139.5, 128.9, 127.8, 126.7, 80.1, 67.1, 57.0, 28.5(3C). IR (cm-1) 3319, 2976, 1672, 1547, 1494, 1366, 1252, 1169, 1055, 701. TLC Rf: (Heptane/EtOAc, 1:1) = 0.45. LCMS (ESI) m/z C13H19NO3 [M+H]+ 238.4. 13
tert-butyl (R)-(2-azido-1-phenylethyl)carbamate (S21) C13H18N4O2 MW: 262.14 g.mol-1 Yield: 54% White solid To a stirred solution of tert-butyl (R)-(2-hydroxy-1-phenylethyl)carbamate S20 (1.26 mmol, 300.0 mg) in 12.0 mL of dry CH2Cl2 at 0 °C, were sequentially added NEt3 (1.39 mmol, 194.0 µL) and MsCl (1.32 mmol, 102.0 µL). The resulting reacting mixture was stirred under argon at RT for 2 hours. After the reaction was completed, the mixture was diluted by addition 20.0 mL of CH2Cl2 and washed with 30.0 mL of a NH4Cl saturated solution. The organic phase was dried over anhydrous MgSO4 and concentrated under vacuum. The obtained crude mesylated compound was then solubilized in 10.0 mL of DMF and Sodium azide (2.50 mmol, 162.5 mg) was added. The resulting reacting mixture was stirred at 80 °C overnight. After the reaction was completed, the mixture was diluted in 50.0 mL of EtOAc and washed trice with 30.0 mL of a NaCl saturated solution. The organic phase was dried over anhydrous MgSO4 and concentrated under vacuum. Purification occurred by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1) afforded the tert-butyl (R)-(2-azido1-phenylethyl)carbamate S21 as a low temperature melting white solid (179 mg, 54%). S13
1
H NMR (400 MHz, CDCl3) δ 7.37 (m, 2H), 7.31 (m, 3H), 5.16 (d, J = 6.4 Hz, 1H); 4.88 (s, 1H), 3.62 (s, 2H), 1.44 (s, 9H). C NMR (100 MHz, CDCl3) δ 155.2, 139.3, 128.9 (2C), 128.1, 126.6 (2C), 80.2, 55.7, 54.2, 28.4 (3C). IR (cm-1) 3380, 2982, 2092, 1680, 1511, 1363, 1292, 1249, 1163, 1043, 1023, 853, 754, 700, 617, 514. LCMS (ESI) m/z C13H18N4O2 [M+H]+ 263.2. 13
(R)-2-azido-1-phenylethan-1-amine (S22) C8H10N4 MW: 162.09 g.mol-1 Yield: 84% yellowish oil To a stirred solution of tert-butyl (R)-(2-azido-1-phenylethyl)carbamate S21 (0.382 mmol, 100.0 mg) in 2.0 mL of dry CH2Cl2 at 0 °C was added 200.0 µL of TFA. The resulting reacting mixture was stirred under argon at room temperature for 2 hours. After the reaction was completed, the mixture was diluted by addition 20.0 mL of CH2Cl2 and washed with 20.0 mL of a NaHCO3 saturated solution. The aqueous phase was extracted twice with 20.0 mL of CH2Cl2, The organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. The (R)-2-azido-1-phenylethan-1amine S22 was obtained as a yellowish oil (52 mg, 84%). 1
H NMR (400 MHz, CDCl3) δ 7.44 – 7.26 (m, 5H), 4.15 (dd, J = 8.3, 4.5 Hz, 1H), 3.54 (dd, J = 12.0, 4.5 Hz, 1H), 3.40 (dd, J = 12.0, 8.3 Hz, 1H). 13 C NMR (100 MHz, CDCl3) δ 142.2, 128.7 (2C), 127.9, 126.5 (2C), 59.1, 55.5. IR (cm-1) 3374, 3029, 2921, 2860, 2094, 1720, 1601, 1492, 1452, 1270, 842, 758, 698, 607, 523. TLC Rf: (DCM/MeOH, 95:5) = 0.30. LCMS (ESI) m/z C8H11N4 [M+H]+ 163.2.
tert-butyl (2-(hydroxymethyl)phenyl)carbamate (S23) C12H17NO3 MW: 223.12 g.mol-1 Yield: 78% Yellowish oil To a stirred solution of (2-aminophenyl)methanol (4.06 mmol, 500.0 mg) in 10.0 mL of dry CH2Cl2 at 0 °C, were sequentially added NEt3 (4.88 mmol, 680.0 µL) and Boc2O (4.26 mmol, 930.0 mg). The resulting reacting mixture was stirred under argon at 0 °C for 72 hours. After the reaction was completed, the mixture was diluted in 50.0 mL of CH2Cl2 and washed twice with 30.0 mL of a NH4Cl saturated solution. The organic phase was dried over anhydrous MgSO4 and concentrated under vacuum. Purification occurred by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1 to 8:2) afforded the tert-butyl (2-(hydroxymethyl)phenyl)carbamate S23 as a yellowish oil (706 mg, 78%). S14
1
H NMR (400 MHz, CDCl3) δ 7.84 (d, J = 8.0 Hz, 1H), 7.76 (s, 1H), 7.33 – 7.27 (td, J = 7.4, 0.8 Hz, 1H), 7.13 (dd, J = 7.4, 1.2 Hz, 1H), 7.02 (td, J = 7.4, 0.8 Hz, 1H), 4.59 (s, 2H), 3.19 (s, 1H), 1.53 (s, 9H). 13 C NMR (100 MHz, CDCl3) δ 153.6, 137.8, 129.3, 128.9, 128.9, 123.2, 121.1, 80.5, 63.9, 28.4 (3C). IR (cm-1) 3355, 2978, 2932, 2877, 1728, 1703, 1591, 1523, 1450, 1367, 1241, 1157, 1052, 1023, 752 TLC Rf: (Heptane/EtOAc, 2:8) = 0.20. LCMS (ESI) m/z C12H18NO3 [M+H]+ 224.3.
tert-butyl (2-(azidomethyl)phenyl)carbamate (S24) C12H16N4O2 MW: 248.13 g.mol-1 Yield: 72% Colorless oil To a stirred solution of tert-butyl (2-(hydroxymethyl)phenyl)carbamate S23 (2.24 mmol, 500 mg) in 20.0 mL of dry CH2Cl2 at 0 °C, were sequentially added NEt3 (2.68 mmol, 374 µL) and MsCl (2.35 mmol, 182 µL). The resulting reacting mixture was stirred under argon at RT for 3 hours. After the reaction was completed, the mixture was diluted by addition 30.0 mL of CH2Cl2 and washed with 30.0 mL of a NH4Cl saturated solution. The aqueous phase was extracted twice with 20.0 mL of dry CH2Cl2. The organic phases were combined, washed with 30.0 mL of a saturated solution of NaCl, dried over anhydrous MgSO4 and concentrated under vacuum. The obtained crude mesylated compound was then solubilized in 15.0 mL of DMF and NaN3 (4.48 mmol, 291 mg) was added. The resulting reacting mixture was stirred at 80 °C overnight. After the reaction was completed, the mixture was diluted in 60.0 mL of EtOAc and washed trice with 30.0 mL of a NaCl saturated solution. The organic phase was dried over anhydrous MgSO4 and concentrated under vacuum. Purification by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 95:5) afforded the tert-butyl (2-(azidomethyl)phenyl)carbamate S24 as a colorless oil (396 mg, 72%). 1
H NMR (400 MHz, CDCl3) δ 7.88 (d, J = 7.9 Hz, 1H), 7.35 (t, J = 7.8 Hz, 1H), 7.22 (d, J = 6.9 Hz, 1H), 7.07 (t, J = 7.4 Hz, 1H), 6.85 (s, 1H), 4.32 (s, 2H), 1.53 (s, 9H). 13 C NMR (100 MHz, CDCl3) δ 153.1, 137.1, 129.9, 129.7, 125.2, 123.8, 122.3, 80.8, 52.8, 28.4 (3C). IR (cm-1) 3404, 2979, 2932, 2098, 1728, 1590, 1519, 1450, 1367, 1297, 1234, 1156, 1049, 753 TLC Rf: (Heptane/EtOAc, 2:8) = 0.45. LCMS (ESI) m/z C12H17N4O2 [M+H]+ 249.3.
2-(azidomethyl)aniline (S25) C7H8N4 MW: 148.07 g.mol-1 Yield: 87% yellowish oil To a stirred solution of tert-butyl (2-(azidomethyl)phenyl)carbamate S24 (0.92 mmol, 230 mg) in 5.0 mL of dry CH2Cl2 at 0 °C was added 500 µL of TFA. The resulting reacting mixture was stirred under argon at RT for 2 hours. After the S15
reaction was completed, the mixture was diluted by addition 30.0 mL of CH2Cl2 and washed with 30.0 mL of a NaHCO3 saturated solution. The aqueous phase was extracted twice with 30.0 mL of CH2Cl2, The organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1) afforded the 2-(azidomethyl)aniline S25 as a yellow oil (119 mg, 87%). 1
H NMR (400 MHz, CDCl3) δ 7.19 (t, J = 7.7 Hz, 1H), 7.12 (d, J = 7.0 Hz, 1H), 6.79 (m, 2H), 4.31 (s, 2H), 3.90 (bs, 2H). C NMR (100 MHz, CDCl3) δ 145.5, 130.5, 130.1, 119.7, 118.5, 116.2, 53.0. IR (cm-1) 3453, 3371, 2096, 1624, 1584, 1495, 1460, 1244, 879, 751. TLC Rf: (Heptane/EtOAc, 2:8) = 0.30. LCMS (ESI) m/z C7H9N4 [M+H]+ 149.2. 13
S16
II. 4. Synthesis of drug precursors.
Synthesis of the 4c precursor
5-(4-(4-chlorophenyl)piperazin-1-yl)pentan-1-ol (S26) C15H23N2OCl MW: 282.81 g.mol-1 Yield: 80% White solid
To a stirred solution of 1-(4-chlorophenyl)piperazine (2.54 mmol, 500 mg) and K2CO3(5.60 mmol, 774 mg) in 10.0 mL of dry CH3CN was added (2.80 mmol, 583 mg) of 5-bromopentan-1-ol (80%) in solution in 5.0 mL of dry CH3CN. The resulting reacting mixture was stirred under argon at 45 °C for 48 hours. After the reaction was completed, the mixture was diluted by addition 30.0 mL of CH2Cl2 and the solid was removed by filtration. The filtrate was dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent CH2Cl2/MeOH 98:2) afforded the 5-(4-(4-chlorophenyl)piperazin-1-yl)pentan-1-ol S26 as a white solid (576 mg, 80%). 1
H NMR (400 MHz, CDCl3) δ 7.20 (d, J = 8.9 Hz, 2H), 6.83 (d, J = 9.0 Hz, 2H), 3.65 (t, J = 6.4 Hz, 2H), 3.23 – 3.18 (m, 4H), 2.69 – 2.63 (m, 4H), 2.50 – 2.43 (m, 2H), 2.28 (brs, 1H), 1.65 – 1.57 (m, 4H), 1.44 (qt, J = 6.7 Hz, 2H). 13 C NMR (100 MHz, CDCl3) δ 149.8, 129.1 (2C), 124.8, 117.1 (2C), 62.8, 58.4, 53.1 (2C), 49.0 (2C), 32.4, 26.2, 23.6. IR (cm-1) 3399, 2932, 2825, 1596, 1496, 1448, 1238, 1144, 1129, 815, 676. TLC Rf: (DCM/MeOH, 9:1) = 0.45. LCMS (ESI) m/z C15H24N2O35Cl [M+H]+ 283.5, C15H24N2O37Cl [M+H]+ 285.3.
S17
5-(4-(4-chlorophenyl)piperazin-1-yl)pentanal (S27) C15H21N2OCl MW: 280.80 g.mol-1 Yield: 43% Yellowish oil
Dry DMSO (288 µL, 4.07 mmol) was added at -78 °C to a solution of oxalyl chloride (145 µL, 1.70 mmol) in 6.0 mL of CH2Cl2 and the resulting mixture was stirred at -78 °C for 30 minutes. Thereafter, the alcohol S26 (200 mg, 0.71 mmol, 1.0 equiv.) in 500 µL of dry CH2Cl2 was added and the reaction mixture was stirred at -78 °C for 1 hour after what Et3N (640 µL, 4.60 mmol) was finally added. The mixture was stirred for 15 minutes at -78 °C then warmed at 0 °C. After addition of 20.0 mL of a saturated aqueous solution of NaHCO3 the immiscible layers were separated by decantation. The aq. phase was extracted twice with CH2Cl2 (20.0 mL). The combined organic layers were washed with brine (20.0 mL) and dried over anhydrous MgSO4. The solvent was removed under reduced pressure and the resulting crude product was purified by flash-chromatography over a silica gel column (eluent CH2Cl2/MeOH 98:2 to 95:5) affording the 5-(4-(4chlorophenyl)piperazin-1-yl)pentanal S27 as a yellowish gummy solid (85 mg, 43%). 1
H NMR (400 MHz, CDCl3) δ 9.74 (s, 1H), 7.19 (d, J = 8.8 Hz, 2H), 6.82 (d, J = 9.0 Hz, 2H), 3.16 (m, 4H), 2.59 (m, 4H), 2.47 (td, J = 7.1, 1.3 Hz, 2H), 2.2 (t, J = 7.0, 2H), 1.68 (m, 2H), 1.56 (m, 2H). 13 C NMR (100 MHz, CDCl3) 202.4, 149.9, 129.0 (2C), 124.6, 117.3 (2C), 58.0, 53.1 (2C), 49.1 (2C), 43.6, 26.2, 20.0. TLC Rf: (DCM/MeOH, 9:1) = 0.40. LCMS (ESI) m/z C16H22N2O35Cl [M+H]+ 281.5; C16H22N2O37Cl [M+H]+ 283.3.
2-azido-N-(5-(4-(4-chlorophenyl)piperazin-1-yl)pentyl)aniline (S28) C21H27N6Cl MW: 398.94 g.mol-1 Yield: 74 % Beige solid
To a solution of aldehyde S27 (80 mg, 0.285 mmol) and 2-azido-aniline (40.2 mg, 0.30 mmol) in freshly dried MeOH at 0 °C was added (65.3 µL, 1.14 mmol) of acetic acid and (46.7 mg, 0.57 mmol) of NaOAc. After 20 minutes, NaBH3CN (18.7 mg, 0.285 mmol) was added and the resulting reaction mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was diluted in 30.0 mL of CH2Cl2 and quenched by addition of a 2M aqueous solution of NaOH. The immiscible layers were separated by decantation. The aq. phase was extracted twice with CH2Cl2 (20.0 mL). The combined organic layers were washed with brine (50.0 mL) and dried over anhydrous MgSO4. The solvent was removed under reduced pressure and the resulting crude product was purified by flash-chromatography over a silica gel column(eluent Hept/EtOAc 9:1 to 8:2) affording the 2-azido-N-(5-(4-(4-chlorophenyl)piperazin-1-yl)pentyl)aniline S28 as a beige powder (84 mg, 74%).
S18
1
H NMR (400 MHz, CDCl3) δ 7.20 (d, J = 9.1 Hz, 2H), 7.06 – 6.99 (m, 2H), 6.85 (d, J = 9.1 Hz, 2H), 6.71 (td, J = 7.8, 1.3 Hz, 1H), 6.60 (m, 1H), 4.00 (brs, 1H), 3.17 (m, 4H), 3.12 (t, J = 6.9 Hz, 2H), 2.60 (m, 4H), 2.41 (m, 2H), 1.67 (qt, J = 6.9 Hz, 2H), 1.58 (m, 2H), 1.45 (m, 2H). 13 C NMR (100 MHz, CDCl3) δ 150.1, 140.1, 129.1 (2C), 126.0, 124.6, 124.5, 117.9, 117.3 (2C), 117.0, 110.9, 58.6, 53.3 (2C), 49.3 (2C), 43.5, 29.4, 26.8, 25.2. IR (cm-1) 3415, 2936, 1818, 2120, 1600, 1517, 1496, 1446, 1288, 1235, 1148, 816, 738. TLC Rf: (Heptane/EtOAc, 1:1) = 0.40. HRMS (ESI) m/z calcd for C21H28N635Cl [M+H]+ : 399.2058; found: 399.2058. Synthesis of the Oxatomide precursor
3-((2-azidophenyl)amino)propan-1-ol (S29) C9H12N4O MW: 192.22 g.mol-1 Yield: 69% brown oil To a stirred solution of 2-azido-aniline 1a (2.23 mmol, 300 mg) in 2.0 mL of dry CH3CN were sequentially added trimethylene oxide (2.46 mmol, 160 µL) and LBF4(2.46 mmol, 230 mg). The resulting reacting mixture was stirred under argon at RT for 30 hours. After the reaction was completed, the mixture was diluted by addition 50.0 mL of CH2Cl2 and washed with 50.0 mL of a NaHCO3 saturated solution. The aqueous phase was extracted with 30.0 mL of CH2Cl2 and the organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1 to 8:2) afforded the 3-((2-azidophenyl)amino)propan-1-ol S29 as a brown oil (295 mg, 69%). 1
H NMR (400 MHz, CDCl3) δ 7.10 – 6.96 (m, 2H), 6.74 (t, J = 7.4 Hz, 1H), 6.65 (d, J = 8.0 Hz, 1H), 3.78 (t, J = 5.9 Hz, 2H), 3.26 (t, J = 6.6 Hz, 2H), 1.88 (qt, J = 6.3 Hz, 2H). 13 C NMR (100 MHz, CDCl3) δ 139.9, 125.9, 124.8, 117.8, 117.3, 111.1, 61.1, 41.2, 31.8. IR (cm-1) 3402, 2939, 2876, 2119, 1601, 1516, 1442, 1288, 1118, 738. TLC Rf: (Heptane/EtOAc, 1:1) = 0.50 LCMS (ESI) m/z C9H13N4O [M+H]+ 193.2; S19
2-azido-N-(3-(4-benzhydrylpiperazin-1-yl)propyl)aniline (S30) C26H30N6 MW: 426.57 g.mol-1 Yield: 40% Yellow oil
To a stirred solution of 3-((2-azidophenyl)amino)propan-1-ol S29 (0.681 mmol, 131 mg) in 7.0 mL of dry CH2Cl2 at 0 °C was added dry Et3N (0.817 mmol, 114 µL) followed by the addition of of methansulfonyl chloride (0.716 mmol, 55.4 µL). The resulting reacting mixture was then stirred at RT for 1 hour. After the reaction was completed, the mixture was diluted by addition 25.0 mL of CH2Cl2 and washed with 25.0 mL of a NH4Cl saturated solution. The aqueous phase was extracted with 20.0 mL of CH2Cl2 and the organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. The crude mesylated compound was then added to a solution of 1-benzhydrylpiperazine (0.817 mmol, 206 mg) and dry Et3N (2.04 mmol, 285 µL) in 3.50 mL of dry DMF. The resulting reaction mixture was stirred under argon at 40°C for 15 hours. After the reaction was completed, the mixture was diluted by addition 50.0 mL of EtOAc and washed trice with 30.0 mL of a NaCl saturated solution. The organic phases was dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent CH2Cl2/EtOAc 9:1) afforded the 2-azido-N-(3-(4-benzhydrylpiperazin-1-yl)propyl)aniline S30 as a yellow oil (115 mg, 40%). 1
H NMR (400 MHz, CDCl3) δ 7.46 (d, J = 7.4 Hz, 4H), 7.30 (t, J = 7.6 Hz, 4H), 7.20 (t, J = 7.3 Hz, 2H), 7.04 (t, J = 7.5 Hz, 2H), 6.72 (m, 1H), 6.60 (d, J = 7.8 Hz, 1H), 5.20 (s, 1H), 4.23 (s, 1H), 3.19 (t, J = 6.2 Hz, 2H), 2.53 (m, 10H), 1.84 (qt, J = 6.2 Hz, 2H). 13 C NMR (100 MHz, CDCl3) δ 143.0 (2C), 140.5, 128.6 (4C), 128.0 (4C), 127.0 (2C), 126.0, 124.5, 118.1, 116.8, 110.7, 76.8, 57.4, 53.7 (2C), 52.1 (2C), 43.3, 25.6. IR (cm-1) 3025, 2943, 2809, 2773, 2117, 1676, 1650, 1600, 1518, 1449, 1292, 1150, 995, 741, 705. TLC Rf: (Heptane/EtOAc, 1:1) = 0.55. LCMS (ESI) m/z C26H31N6 [M+H]+ 427.5. HRMS (ESI) m/z calcd for C26H31N6 [M+H]+ : 427.2605; found: 427.2604.
S20
Synthesis of the Flibanserin precursor
3-((2-azidophenyl)amino)ethan-1-ol (S31) C8H10N4O MW: 178.09 g.mol-1 Yield: 72% Brown oil To a stirred solution of 2-azido-aniline 1a (2.23 mmol, 300 mg) in 2.5 mL of dry CH3CN was added 800 µL of an ethylene oxide solution in THF (2.50 to 3.0 M) and LiBF4 (2.30 mmol, 215 mg). (NB: The volume of ethylene oxide solution might change depending of the actual concentration in ethylene oxide). The resulting reacting mixture was stirred under argon at room temperature for 72 hours. After the reaction was completed, the mixture was diluted by addition 50.0 mL of CH2Cl2 and washed with 50.0 mL of a NaHCO3 saturated solution. The aqueous phase was extracted with 30.0 mL of CH2Cl2 and the organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent Hept/EtOAc 9:1 to 8:2) afforded the 3-((2azidophenyl)amino)ethan-1-ol S31 as a brown oil (289 mg, 57%). 1
H NMR (400 MHz, CDCl3) δ 7.03 (m, 2H), 6.76 (t, J = 7.3 Hz, 1H), 6.65 (d, J = 8.0 Hz, 1H), 3.82 (t, J = 5.2 Hz, 2H), 3.30 (t, J = 5.2 Hz, 2H). 13 C NMR (100 MHz, CDCl3) δ 139.8, 125.9, 125.2, 118.0, 117.8, 111.3, 61.2, 45.8. IR (cm-1) 3400, 2940, 2879, 2123, 1601, 1516, 1443, 1291, 1150, 1044, 739 TLC Rf: (Heptane/AeOAc, 1:1) = 0.50. LCMS (ESI) m/z C8H11N4O [M+H]+ 179.2. 2-azido-N-(2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)aniline (S32) C19H21F3N6 MW: 390.18 g.mol-1 Yield: 40% yellow oil
S21
To a stirred solution of 3-((2-azidophenyl)amino)ethan-1-ol S31 (0.736 mmol, 131 mg) in 7.50 mL of dry CH2Cl2 at 0 °C was added dry Et3N (0.90 mmol, 127 µL) followed by the addition of MsCl (0.750 mmol, 58.0 µL). The resulting reacting mixture was then stirred at 0 °C for 20 minutes. After the reaction was completed, the mixture was diluted by addition 25.0 mL of CH2Cl2 and washed with 25.0 mL of a NH4Cl saturated solution. The aqueous phase was extracted with 20.0 mL of CH2Cl2 and the organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. The crude mesylated compound was then added to a solution of 1-(4-(trifluoromethyl)benzyl)piperazine hydrochloride (0.736 mmol, 200 mg) and dry Et3N (1.54 mmol, 216 µL) in 5.0 mL of dry DMF. The resulting reaction mixture was stirred under argon at 45 °C for 15 hours. After the reaction was completed, the mixture was diluted by addition 50.0 mL of EtOAc and washed trice with 30.0 mL of a NaCl saturated solution. The organic phases was dried over anhydrous MgSO4 and concentrated under vacuum. Purification performed by Flash Chromatography on SiO2 gel (eluent Heptane/EtOAc 9:1 to 8:2) afforded the 2-azido-N-(2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)aniline S32 as a brownish oil (54 mg, 19%). 1
H NMR (400 MHz, CDCl3) δ 7.36 (t, J = 8.0 Hz, 1H), 7.15 (s, 1H), 7.11 – 7.03 (m, 4H), 6.76 (td, J = 7.7, 1.3 Hz, 1H), 6.64 (dd, J = 8.0, 1.1 Hz, 1H), 4.69 (s, 1H), 3.31 – 3.28 (m, 4H), 3.23 (t, J = 6.0 Hz, 2H), 2.74 (t, J = 6.0 Hz, 2H), 2.69 – 2.65 (m, 4H). 13 C NMR (100 MHz, CDCl3) δ 151.5, 140.1, 131.5 (q, J = 32 Hz), 129.7, 126.0, 125.0, 124.4 (q, J = 271 Hz), 118.8, 118.0, 117.3, 115.9 (q, J = 3.8 Hz), 112.3 (q, J = 3.9 Hz), 111.2, 56.5, 52.7 (2C), 48.8 (2C), 40.1 ppm. 19 F NMR (376 MHz, CDCl3) δ 62.62. TLC Rf: (Heptane/EtOAc, 8:2) = 0.20. IR (cm-1) 2944, 2881, 2823, 2120, 1601, 1506, 1448, 1315, 1288, 962, 739. LCMS (ESI) m/z C19H22F3N6 [M+H]+ 391.5. HRMS (ESI) m/z calcd for C19H22F3N6 [M+H]+ : 363.1791; found: 363.1786.
Synthesis of the Domperidone precursor
S22
1-(1-(3-((2-azidophenyl)amino)propyl)piperidin-4-yl)-5-chloro-1,3-dihydro-2H-benzo[d]imidazol-2-one (S33) C21H24ClN7O MW: 425.92 g.mol-1 Yield: 42% Brown solid
To a stirred solution of 3-((2-azidophenyl)amino)ethan-1-ol (0. 624 mmol, 120 mg) in 6.2 mL of dry CH2Cl2 at 0 °C was added dry Et3N (0.75 mmol, 104 µL) followed by the addition of of MsCl (0.624 mmol, 48.0 µL). The resulting reacting mixture was then stirred at 0 °C for 20 minutes. After the reaction was completed, the mixture was diluted by addition 20.0 mL of CH2Cl2 and washed with 25.0 mL of a NH4Cl saturated solution. The aqueous phase was extracted with 20.0 mL of CH2Cl2 and the organic phases were combined, dried over anhydrous MgSO4 and concentrated under vacuum. The crude mesylated compound S29 was then added to a stirred solution of 5-chloro-1-(piperidin-4-yl)-1,3-dihydro-2Hbenzo[d]imidazol-2-one (0.624 mmol, 157 mg) in 6.2 mL of dry DMF. The resulting mixture was heated under argon at 40 °C for 3 days then DMF was evaporated under vacuum. The crude mixture was extracted with 30.0 mL of EtOAc and the organic phases were combined, dried over anhydrous MgSO4 and concentrated. Purification performed by Flash Chromatography on SiO2 gel (cHex from 30% to 100% EtOAc) afforded the 1-(1-(3-((2azidophenyl)amino)propyl)piperidin-4-yl)-5-chloro-1,3-dihydro-2H-benzo[d]imidazol-2-one S33 as brown solid (110 mg, 42%). 1
H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.07 (dd, J = 7.7, 1.3 Hz, 1H), 7.03 (dd, J = 8.4, 2.0 Hz, 2H), 7.00 (d, J = 2.0 Hz, 1H), 6.65 (m, 2H), 5.42 (t, J = 5.3 Hz, 1H), 4.14 (ddd, J = 12.3, 8.3, 4.2 Hz, 1H), 3.13 (m, 2H), 3.05 (br. d, J = 11.4 Hz, 2H), 2.43 (t, J = 6.3 Hz, 2H), 2.40 – 2.32 (m, 2H), 2.02 (t, J = 11.7 Hz, 2H), 1.76 – 1.72 (m, 2H), 1.68 (br. d, J = 9.8 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 153.6, 140.2, 129.5, 128.2, 126.0, 124.7, 123.5, 120.0, 118.2, 116.1, 110.4, 109.5, 108.6, 56.3, 52.9 (2C), 50.5, 42.0, 28.5 (2C), 25.4. TLC Rf: (cHex/EtOAc, 1:9) = 0.20. IR (cm-1) 2929, 2120, 1697, 1600, 1486, 1371, 1289, 1151, 741. LCMS (ESI) m/z C21H24ClN7O [M+H]+ 428.3 [37Cl], 426.4 [35Cl]. HRMS (ESI) m/z calcd for C21H24ClN7O 6 [M+H]+ 426.1802; found: 426.1803.
S23
SDWSTEK (S34) C39H57N13O17 MW: 963.96 g.mol-1 Yield: 21%
The peptide SDWSTEK S34 was synthesized by Fmoc-SPPS on a Prelude synthesizer from Protein technologies (6 x 25 mmol). It was produced on a Fmoc-Lys(Boc)-Wang resin with N-methyl pyrolidone (NMP) for solvent and 20% piperidine in NMP for Fmoc-deprotection. 2-(6-Chloro-1-H-benzotriazole-1-yl)-1,1,3,3-tetramethylammonium hexafluorophosphate (HCTU) was used as condensation reagent in presence of N-methyl morpholine. After each coupling the growing peptide was capped with acetic anhydride. At the end of the synthesis, the resin was assembled and coupled in a syringe with Boc-3-azido-Ala-OH (0.75 mmol, 137 mg) in 4 ml of DMF, in presence of N,N-dimethylmethaneiminium hexafluorophosphate (HATU) (0,75 mmol, 285 mg) and DIEA (1.5 mmol, 260 ml), and was shaken for 3 hours. Three washings with NMP and two washings with CH2Cl2 were carried out. The peptide was deprotected in a mixture of TFA/TIS/H2O/phenol (8.8/0.5/0.5/0.2) for 2 hours, precipitated in ethyl ether and checked by reverse phase HPLC (see below; the chromatogram was taken at 214 nm on a column Vydac C18, 10x250 mm, 10 mM A: H2O, 0.1% TFA, B: CH3CN; 5 to 20% B in 20 min). The peptide S34 was used without further purification.
S24
II. 5. Preparation of [13C]benzo[d]imidazol-2(3H)-one and [13C]tetrahydropyrimidin-2(1H)-one derivatives General procedure for the synthesis of benzo[d]imidazol-2(3H)-one from the corresponding aromatic/aliphatic N3derivatives:
In to 1.0 mL vial, PPhMe2 (1.00 equiv.) was added to a solution of amino-azide derivative (1.00 equiv.) in the appropriate solvent (0.50 mL). The solution was transferred into a Wilmad® low pressure/vacuum NMR tube that was further freezed in to N2 bath, then 1.0 to 1.50 equiv. of gaseous 13CO2 was added using Tritec® (figure 1). The mixture was maintained RT for 5 to 10 minutes then the unreacted 13CO2 was removed by opening the NMR tube and the solvent was evaporated. The crude products were purified by Flash Chromatography on SiO2 gel or Preparative TLC, affording the corresponding [13C]benzo[d]imidazol-2(3H)-one.
II. A. Optimization of the Staudinger-Aza-Wittig reaction (SAW) Entry
Solvent
Phosphine
Temperature
Time
Yield
1
TDF
PPh3
65°C
2
79
2
MeOD
PPh3
65°C
2
92
3
CD3CN
PPh3
65°C
2
>90
4
CD3CN
PPh3
RT
2
80
5
CD3CN
PPh2Me
65°C
2
>95
6
CD3CN
PPh2Me
65°C
1h
>95
7
CD3CN
PPh2Me
65°C
30 min
70
8
CD3CN
PPh2Me
RT
2h
>95
9
CD3CN
PPh2Me
RT
1h
84
11
CD3CN
PPhMe2
RT
1h
>95
12
CD3CN
PPhMe2
RT
30 min
>95
S25
13
CD3CN
PPhMe2
RT
15 min
>95
14
CD3CN
PPhMe2
RT
5 min
>95
Solvent screening
Entry
Solvent
Time
Temperature
Yield
1
CD3CN
5 min
RT
>95
2
TDF
5 min
RT
35
3
MeOH-d4
5 min
RT
85
4
DMF-d7
5 min
RT
>95
5
DMSO-d6
5 min
RT
87
6
Toluene-d8
5 min
RT
75
figure 1: Tritec®
S26
[13C] 1H-benzo[d]imidazol-2(3H)-one (13C-2a) C613CH6N2O MW: 135.13 g.mol-1 Yield: 90% White solid The [13C] 1H-benzo[d]imidazol-2(3H)-one 13C-2a was prepared accordingly to the general procedure, using PPhMe2 (0.108 mmol, 16.4 µL), 2-azidoaniline (0.108 mmol, 14.6 mg) and 13CO2 (0.120 mmol). The crude product was purified by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 1H-benzo[d]imidazol-2(3H)-one 13C-2a as a white solid (14.5 mg, 90%). The spectral data (1H-NMR) was consistent with reported one.6 1
H NMR (400 MHz, DMSO-d6) δ 6.97 (m, 4H). C NMR (100 MHz, MeOD) δ 158.1, 130.8 (d, J = 4.9 Hz, 2C), 122.5 (2C), 110.3 (d, J = 4.2 Hz, 2C). IR (cm-1) 3106, 3012, 2894, 2806, 1685, 1482, 1190, 729, 699, 592. LCMS (ESI) m/z C613CH6N2O [M+H]+ 136.0. 13
[13C] 5-chloro-1H-benzo[d]imidazol-2(3H)-one (13C-2b) C613CH5ClN2O MW: 169.57 g.mol-1 Yield: 71% White solid The [13C] 5-chloro-1H-benzo[d]imidazol-2(3H)-one 13C-2b was prepared accordingly to the general procedure, using PPhMe2 (0.098 mmol, 16.4 µL), 2-azido-4-chloroaniline (0.09 mmol; 16.6 mg) and 13CO2 (0.120 mmol). The crude product was purified with FC on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 5-chloro-1H-benzo[d]imidazol-2(3H)-one 13C2b as a white solid (12 mg, 71%). The spectral data (1H-NMR) was consistent with reported one.7 1
H NMR (400 MHz, DMSO-d6) δ 10.77 (s, 2H), (m, 2H), 6.97-6.89 (m, 3H). C NMR (100 MHz, DMSO-d6) δ 155.2, 130.8 (d, J = 4.9 Hz), 128.6 (d, J = 4.9 Hz), 124.4, 120.1, 109.5 (d, J = 4.2 Hz), 108.3 (d, J = 4.3 Hz). IR (cm-1) 3407, 3154, 2921, 1716, 1604, 1655, 1487, 1191, 795, 717, 597. LCMS (ESI) m/z C613CH5ClN2O [M+H]+ 170.1 13
[13C] 5-fluoro-1H-benzo[d]imidazol-2(3H)-one (13C-2c) C613CH5FN2O MW: 153.12 g.mol-1 Yield: 65% White solid The [13C] 5-fluoro-1H-benzo[d]imidazol-2(3H)-one 13C-2c was prepared accordingly to the general procedure, using 2azido-4-fluoroaniline (18.2 mg, 0.120 mmol), PPhMe2 (17.0 μL, 0.120 mmol) and 13CO2 (0.120 mmol). After 5 minute, the crude product was purified by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:1 then DCM/MeOH 95:5)
6
Dai, L.-L.; Zhang, H.-Z.; Nagarajan, S.; Rasheed, S.; Zhou, C.-H. Med. Chem. Commun. 2015, 6, 147.
S27
affording the [13C] 5-fluoro-1H-benzo[d]imidazol-2(3H)-one 13C-2c as a white solid (12 mg, 65%). The spectral data (1HNMR) was consistent with reported one. 1
H NMR (400 MHz, DMSO-d6) δ 10.75 (s, 1H), 10.63 (s, 1H), 6.89-6.86 (m, 1H), 6.78–6.71 (m, 2H). C NMR (100 MHz, CDCl3) δ 156.7, 156.1, 130.9, 130.7, 109.1, 107.0, 106.8. IR (cm-1) 3089, 2959, 1634, 1615, 1500, 1476, 1140, 780, 709, 597, 575. LCMS (ESI) m/z C613CH5FN2O[M+H]+ 154.2. 13
[13C] 5-iodo-1H-benzo[d]imidazol-2(3H)-one (13C-2d) C613CH5IN2O MW: 261.03 g.mol-1 Yield: 37% White solid The [13C]5-iodo-1H-benzo[d]imidazol-2(3H)-one 13C-2d was prepared accordingly to the general procedure, using 2azido-4-iodoaniline (26.0 mg, 0.10 mmol), PPhMe2 (13.5 μL, 0.10 mmol) and 13CO2 (0.10 mmol). The crude product was purified with FC on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C]5-iodo-1H-benzo[d]imidazol-2(3H)-one 13C-2d as a white solid (14.5 mg, 37%). 1
H NMR (400 MHz, DMSO-d6) δ 10.74 (dd, J = 13.7, 3.7 Hz, 2H), 7.24 (dd, J = 8.1, 1.6 Hz, 1H), 7.19 (d, J = 1.4 Hz, 1H), 6.76 (d, J = 8.1 Hz, 1H). 13 C (100 MHz, DMSO-d6) δ 154.7, 131.5 (d, J = 4.9 Hz), 129.4 (d, J = 4.9 Hz), 128.7, 116.4 (d, J = 4.3 Hz), 110.7 (d, J = 4.2 Hz), 82.7 (s). IR (cm-1): 3139, 3047, 2976, 2819, 2862, 169, 1650, 1622, 1597, 1473, 1342, 1189, 1022, 1004, 789, 764, 708, 591, 580, 532. LCMS (ESI) m/z C9H10N2 [M+H]+ 262.2. [13C] 5-methoxy-1H-benzo[d]imidazol-2(3H)-one (13C-2e) C713CH8N2O2 MW: 165.16 g.mol-1 Yield: 78% White solid The [13C] 5-methoxy-1H-benzo[d]imidazol-2(3H)-one 13C-2e was prepared accordingly to the general procedure, using 2azido-4-methoxyaniline (18.3 mg, 0.112 mmol), PPhMe2 (16 μL, 0.112 mmol) and 13CO2 (0.112 mmol). The crude product was purified by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 5-methoxy-1Hbenzo[d]imidazol-2(3H)-one 13C-2e as a white solid (14.5 mg, 78%). 1
H NMR (400 MHz, DMSO- d6) δ 10.52 (s, 1H), 10.39 (s, 1H), 6.80 (d, J = 8.3 Hz, 1H), 6.50 (d, J = 10.3 Hz, 2H), 3.69 (s, 3H). C NMR (100 MHz, DMSO- d6) δ 156.1, 154.8, 130.9 (d, J = 4.7 Hz), 124 (d, J = 4.6 Hz), 109.1 (d, J = 0.4 Hz), 106.5, 95.7 (d, J = 3.8 Hz), 55.9. IR (cm-1) 3074, 2999, 2596, 2891, 2798, 1707, 1659, 1504, 1487, 1462, 1344, 1193, 1154, 1023, 169, 707, 603, 578. LCMS (ESI) m/z C713CH8N2O2 [M+H+] 166.1. 13
S28
[13C] tert-butyl (2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate (13C-2f) C1113CH15N3O3 MW: 250.26 g.mol-1 Yield: 73 % Red Solid The [13C]tert-butyl (2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate 13C-2f was prepared accordingly to the general procedure, using tert-butyl (4-amino-3-azidophenyl)carbamate (21.2 mg, 0.085 mmol), PPhMe2 (12.1 μL, 0.085 mmol) and 13CO2 (0.090 mmol). The crude was purified with Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9 then DCM/MeOH 95:5) affording the [13C]tert-butyl (2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate 13C-2f as a red solid (15.3 mg, 73%). 1
H NMR (400 MHz, DMSO-d6) δ 10.48 (d, J = 3.7 Hz, 1H), 10.40 (d, J = 3.4 Hz, 1H), 9.15 (s, 1H), 7.22 (s, 1H), 6.92 (d, J = 7.3 Hz, 1H), 6.77 (d, J = 8.4 Hz, 1H), 1.46 (s, 9H). 13 C NMR (100 MHz, DMSO-d6) δ 156.2, 154.6 (m), 153.4, 151.5, 133.6, 130.2 (d, J = 5.1 Hz), 125.2 (d, J = 4.7 Hz), 111.4 (d, J = 5.7 Hz), 108.6 (d, J = 3.9 Hz), 100.2 (d, J = 6.0 Hz), 79.1, 28.6. IR (cm-1) 3345, 3148, 2983, 2929, 2800, 1691, 1656, 1527, 1512, 1236, 1164, 1053, 1023, 797, 773, 761, 711, 640, 601. LCMS (ESI) m/z C1113C H15N3O3 [M+H]+ 251.2.
[13C] methyl 2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-carboxylate (13C-2g) C813CH8N2O3 MW: 193.16 g.mol-1 Yield: 40% White solid The [13C] 5-methoxy-1H-benzo[d]imidazol-2(3H)-one 13C-2g was prepared accordingly to the general procedure, using methyl 4-amino-3-azidobenzoate (25.6 mg, 0.133 mmol), PPhMe2 (19.0 μL, 0.133 mmol) and 13CO2 (0.133 mmol). The crude was purified by Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 5-methoxy-1Hbenzo[d]imidazol-2(3H)-one 13C-2g as a white solid (10.2 mg, 40%). 1
H NMR (400 MHz, DMSO-d6) δ 11.05 (br. s, 1H), 10.89 (d, J = 3.3 Hz, 1H), 7.63 (dd, J = 8.2, 1.3 Hz, 1H), 7.47 (s, 1H), 7.02 (d, J = 8.2 Hz, 1H), 3.81 (s, 3H). 13 C NMR (100 MHz, DMSO-d6) δ 166.8, 155.8 (m), 134.4 (d, J = 4.8 Hz), 130.1 (d, J = 5.0 Hz), 123.3, 122.2, 109.4 (d, J = 4.5 Hz), 108.6 (d, J = 4.2 Hz), 52.3. IR (cm-1): 3137, 3077, 3961, 3922, 3851, 1723, 1632, 1434, 1298, 1270, 1223, 1081, 755, 738, 710, 600, 551. LCMS (ESI) m/z C813CH8N2O3[M+H]+ 194.1.
S29
[13C] 5-(trifluoromethyl)-1H-benzo[d]imidazol-2(3H)-one (13C-2h) C713CH5F3N2O MW: 203.13 g.mol-1 Yield: 27% White solid The [13C] 5-(trifluoromethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2h was prepared accordingly to the general procedure, using 2-azido-4-(trifluoromethyl)aniline (11 mg, 0.054 mmol), PPhMe2 (7.7 μL, 0.054 mmol) and 13CO2 (0.055 mmol). Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 1/9 to DCM : MeOH 95/5) afforded the [13C] 5(trifluoromethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2h as a white solid (3 mg, 27%). 1
H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 10.96 (s, 1H), 7.29 (d, J = 8.2 Hz, 1H), 7.16 (s, 1H), 7.09 (d, J = 8.1 Hz, 1H). C NMR (100 MHz, DMSO-d6) δ 155.7, 129.1, 128.9, 126.2, 125.4, 123.6, 121.1, 120.9. IR (cm-1) 3078, 2956, 2921, 2851, 2747, 1698, 1633, 1651, 1611, 1313, 1239, 1168, 1112, 1050, 1022, 818, 711, 659, 601, 568, 541. LCMS (ESI) m/z C713CH5F3N2O [M+H]+ 205.6. 13
[13C] 1H-naphtho[1,2-d]imidazol-2(3H)-one (13C-2i) C1013CH8N2O3 MW: 185.19 g.mol-1 Yield: 92% Beige solid The [13C] 1H-naphtho[1,2-d]imidazol-2(3H)-one 13C-2i was prepared accordingly to the general procedure, using 2azidonaphthalen-1-amine (21.5 mg, 0.139 mmol), PPhMe2 (19.8 μL, 0.139 mmol) and 13CO2 (0.140 mmol). Purification by Flash Chromatography on SiO2 gel (eluent cHex : EtOAc 1/9) afforded the [13C] 1H-naphtho[1,2-d]imidazol-2(3H)-one 13C2i as a Beige solid (19.8 mg, 92%). 1
H NMR (400 MHz, DMSO-d6) δ 11.47 (s, 1H), 10.88 (d, J = 2.9 Hz, 1H), 8.03 (d, J = 8.3 Hz, 1H), 7.87 (d, J = 8.2 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.48 (t, J = 7.3 Hz, 1H), 7.34 (t, J = 7.2 Hz, 1H), 7.27 (d, J = 8.5 Hz, 1H). 13 C NMR (100 MHz, DMSO-d6) δ 156.0 – 155.5 (m), 129.1, 128.9, 126.2, 125.4 (d, J = 4.9 Hz), 123.6, 123.5 (d, J = 5.1 Hz), 121.1, 120.9, 119.5 (d, J = 3.9 Hz), 111.10 (d, J = 4.2 Hz). IR (cm-1) 3136, 3106, 3034, 2957, 2852, 2668, 1681, 1644, 1620, 1587, 1386, 1183, 1012, 796, 773, 656, 605, 588, 556. LCMS (ESI) m/z C1013C H8N2O3 [M+H]+ 186.1.
S30
[13C] 4,6-dimethyl-1H-benzo[d]imidazol-2(3H)-one (13C-2j) C813CH10N2O MW: 163.18 g.mol-1 Yield: 79% White solid The [13C] 4,6-dimethyl-1H-benzo[d]imidazol-2(3H)-one 13C-2j was prepared accordingly to the general procedure, using 2-azido-4,6-dimethylaniline (18.3 mg, 0.112 mmol), PPhMe2 (16.0 μL, 0.112 mmol) and 13CO2 (0.112 mmol). The crude product was purified with Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 4,6-dimethyl1H-benzo[d]imidazol-2(3H)-one 13C-2j as a White solid (14.5 mg, 79%). 1
H NMR (400 MHz, DMSO-d6) δ 10.53 (s, 1H), 10.44 (s, 1H), 6.55 (s, 1H), 6.54 (s, 1H), 2.23 (s, 3H), 2.20 (s, 3H). C NMR (100 MHz, DMSO-d6) δ 156, 129.8 (d, J = 5.0 Hz), 129.7, 126.8 (d, J = 5.2 Hz), 122.7, 118.2 (d, J = 3.6 Hz), 107 (d, J = 3.7 Hz), 21.4, 16.5. IR (cm-1) 3136, 3003, 2796, 2915, 2851, 1685, 1641, 1617, 1510, 1468, 1301, 1217, 1012, 829, 716, 617, 593, 541. LCMS (ESI) m/z C613CH10N2O+ [M-H+] 164.3. 13
[13C] 4-phenyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (13C-2k) C1213CH10N2O MW: 211.08 g.mol-1 Yield: 85% Beige solid
The [13C] 4-phenyl-1,3-dihydro-2H-benzo[d]imidazol-2-one 13C-2k was prepared accordingly to the general procedure, using PPhMe2 (0.10 mmol, 14.4 µL), 3-azido-[1-1’-biphenyl]-2-amine (0.10 mmol, 21.6 mg) and 13CO2 (0.120 mmol). The crude product was purified with Flash Chromatography on SiO2 gel (eluent cHex/EtOAc 1:9) affording the [13C] 4-phenyl1,3-dihydro-2H-benzo[d]imidazol-2-one 13C-2k as a beige solid (17.3 mg, 85%). The spectral data (1H-NMR) was consistent with reported one.8 1
H NMR (400 MHz, MeOD) δ 7.55 (m, 2H), 7.51-7.47 (m, 2H), 7.15-7.11 (m, 1H), 7.08-7.04 (m, 2H). C NMR (100 MHz, MeOD) δ 158.33, 139.1, 131.3, 130 (3C), 129.2 (3C), 128.6, 125.6, 122.9, 109.4. IR (cm-1) 3111.3, 2915.5, 1647.9, 1614.2, 1432.2, 748.4, 737.4, 693.1, 650.5, 600. LCMS (ESI) m/z C613CH5ClN2O [M+H]+ 212.3. 13
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[13C] 1-benzyl-1H-benzo[d]imidazol-2(3H)-one (13C-2l) C1313CH12N2O MW: 225.25 g.mol-1 Yield: 53 % Dark brown solid
[13C] 1-benzyl-1H-benzo[d]imidazol-2(3H)-one 13C-2l was prepared accordingly to the general procedure, using 2-azidoN-benzylaniline (29.9 mg, 0.133 mmol), PPhMe2 (17.8 μL, 0.133 mmol) and 13CO2 (0.133 mmol). Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 1/9 to DCM : MeOH 95/5) afforded the [13C] 1-benzyl-1Hbenzo[d]imidazol-2(3H)-one 13C-2l as a dark brown solid (16 mg, 53%). 1
H NMR (400 MHz, DMSO-d6) δ 10.97 (d, J = 3.5 Hz, 1H), 7.38 – 7.10 (m, 5H), 7.07 – 6.88 (m, 4H), 5.00 (d, J = 3.7 Hz, 2H).
13
C NMR (100 MHz, DMSO-d6) δ 154.3, 137.2, 129.9 (d, J = 6.6 Hz), 128.5 (2C), 128.2 (d, J = 4.0 Hz), 127.3, 127.2 (2C), 121.0, 120.5, 108.8, 108.1, 43.1. IR (cm-1) 3154, 3024, 2922, 1638, 1261, 1482, 1381, 1348, 726, 746, 696, 681, 626, 557. LCMS (ESI) m/z C13H12N4 [M+H]+ 226.2.
[13C] 1-(naphthalen-2-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (13C-2m) C1713CH14N2O MW: 275.31 g.mol-1 Yield: 65% White solid
The [13C]1-(naphthalen-2-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2m was prepared accordingly to the general procedure, using 2-azido-N-(naphthalen-2-ylmethyl)aniline (27.4 mg, 0.10 mmol), PPhMe2 (14.2 μL, 0.10 mmol) and 13 CO2 (0.10 mmol). Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 1/9) afforded the [13C]1(naphthalen-2-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2m as a white solid (17.9 mg, 65%). 1
H NMR (400 MHz, DMSO-d6) δ 11.00 (d, J = 4.6 Hz, 1H), 7.88-7.84 (m, 4H), 7.51-7.44 (m, 3H), 7.06-6.9 (m, 4H), 5.17 (d, J= 3.7 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 154.8, 135.2, 133.2, 132.7, 130.5, 130.4, 128.8, 128.7, 128.1, 128.0, 126.8, 126.4, 125.9, 121.4, 121.0, 109.3, 108.5, 43.9. IR (cm-1) 3150, 3046, 2931, 1638, 1481, 7445, 689, 723, 486, 471. LCMS (ESI) m/z C1713CH14N2O [M+H]+ 276.4
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[13C] 1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (13C-2n) C1413CH12N2O3 MW: 269.26 g.mol-1 Yield: 78% White solid
The [13C] 1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2n was prepared accordingly to the general procedure, using 2-azido-N-(benzo[d][1,3]dioxol-5-ylmethyl)aniline (26.8 mg, 0.10 mmol), PPhMe2 (14.2 μL, 0.10 mmol) and 13CO2 (0.10 mmol). Purification by Flash Chromatography on SiO2 gel (eluent cHex : EtOAc 1/9) afforded the 21 mg of the [13C] 1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one 13C-2n as a white solid (20.5 mg, 78%). 1
H NMR (400 MHz, DMSO-d6) δ 10.93 (d, J = 4.4 Hz, 1H), 6.99-6.65 (m, 4H), 6.83 (d, J = 7 Hz, 1H), 6.77 (t, J = 7.8 Hz, 1H), 6.64 (d, J = 7.7 Hz, 1H), 6.05 (s, 2H), 4.94 (d, J = 3.7 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 154.8– 154.5 (m), 154.4, 147.50, 145.36, 130.4 (d, J = 6.3 Hz), 128.7 (d, J = 4.5 Hz), 122.2, 121.4 (d, J = 11.0 Hz), 121.1, 118.7, 109.3 (d, J = 4.4 Hz), 108.3, 108.2, 101.5, 38.2. IR (cm-1) 3146, 3059, 2801, 2773, 2680, 2631, 1650, 1634, 1644, 1491, 1461, 1066, 926, 689, 532, 589. LCMS (ESI) m/z C1413CH12N2O3 [M+H]+ 270.2
[13C] 1-benzyltetrahydropyrimidin-2(1H)-one (13C-3a) C1013CH14N2O MW: 192.23 g.mol-1 Yield: 70% White solid The [13C] 1-benzyltetrahydropyrimidin-2(1H)-one 13C-3a was prepared accordingly to the general procedure, using 3azido-N-benzylpropan-1-amine (25.0 mg, 0.130 mmol), PPhMe2 (18.7 μL, 0.130 mmol) and 13CO2 (0.131 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 1/9 to DCM : MeOH 95/5) afforded the [13C] 1-benzyltetrahydropyrimidin-2(1H)-one 13C-3a as a white solid (17.5 mg, 70%). 1
H NMR (400 MHz, DMSO-d6) δ 7.32 (t, J = 7.3 Hz, 2H), 7.23 (t, J = 8.0 Hz, 3H), 6.29 (s, 1H), 4.41-4.4 (d, J = 3.6 Hz, 2H), 3.13-3.08 (m, 4H), 1.79-1.74 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 155.9, 139.3, 128.7 (2C), 127.7 (2C), 127.2, 50.1, 45.1 (2C), 22.5 IR (cm-1) 3250, 3176, 3024, 2931, 2850, 1612, 1503, 1495, 1442, 1287, 1209, 1173, 1013, 730, 721, 695, 676, 485. LCMS (ESI) m/z C1013CH14N2O [M+H]+ 193.5.
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[13C] 1-(thiophen-2-ylmethyl)tetrahydropyrimidin-2(1H)-one (13C-3b) C813CH12N2OS MW: 197.07 g.mol-1 Yield: 68% Beige solid The [13C] 1-(thiophen-2-ylmethyl)tetrahydropyrimidin-2(1H)-one 13C-3b was prepared accordingly to the general procedure, using 3-azido-N-(thiophen-2-ylmethyl)propan-1-amine (21.9 mg, 0.110 mmol), PPhMe2 (15.8 μL, 0.110 mmol) and 13CO2 (0.110 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 (from cHex : EtOAc 1/9 to DCM : MeOH 95/5) afforded the [13C] 1-(thiophen-2-ylmethyl)tetrahydropyrimidin-2(1H)-one 13C-3b as a beige solid (15 mg, 68%). 1
H NMR (400 MHz, DMSO-d6) δ 7.40 (dd, J = 4.9 Hz, 1.3, 1H), 6.95 (m, 2H), 6.33 (s, 1H), 4.55 (d, J = 3.5 Hz, 2H), 3.15 (m, 2H), 3.08 (ddd, J = 8.3, 5.5, 2.6 Hz, 2H), 1.81-1.70 (m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 155.5, 142.1, 126.9, 126.4, 125.8, 114.9, 45.2, 44.8, 22.4. IR (cm-1) 3252, 3180, 3093, 3023, 2961, 2936, 2860, 1613, 1498, 1444, 1276, 1205, 729, 747, 509, 490. LCMS (ESI) m/z C813CH12N2OS [M+H]+ 198.3.
[13C] 1-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)tetrahydropyrimidin-2(1H)-one (13C-3c) C1213C H15N3OS2 MW: 294.4 g.mol-1 Yield: 76% White powder The [13C] 1-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)tetrahydropyrimidin-2(1H)-one 13C-3c was prepared accordingly to the general procedure, using 3-azido-N-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)propan-1-amine (29.3 mg, 0.10 mmol), PPhMe2 (14.2 μL, 0.10 mmol) and 13CO2 (0.10 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (DCM : MeOH 95/5) afforded the [13C] 1-((5-(2-methylthiazol-4-yl)thiophen-2yl)methyl)tetrahydropyrimidin-2(1H)-one 13C-3c as a white solid (22.3 mg, 76%). 1
H NMR (400 MHz, CD3OD) δ 7.43 (s, 1H), 7.31 (d, J = 3.6 Hz, 1H), 6.95 (d, J = 3.6 Hz, 1H), 4.65 (d, J = 3.2 Hz, 2H), 3.34 – 3.32 (m, 2H), 3.26 (td, J = 5.9, 3.1 Hz, 2H), 2.71 (s, 3H), 1.90 (dd, J = 11.7, 5.8 Hz, 2H). 13 C NMR (100 MHz, CD3OD) δ 168.4, 158.3, 150.5, 142.2, 139, 128.0, 124.8, 112.7, 46.7 (d, J= 1.01 Hz) 46.0, 41.1, 23.1 (d, J= 3.03 Hz), 18.7. IR (cm-1) 3265, 2925, 2854, 1611, 1495, 1444, 1292, 1205, 1167, 813, 730, 681. LCMS (ESI) m/z C1213CH15N3OS2 [M+H]+ 295.3.
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[13C] 3,4-dihydroquinazolin-2(1H)-one (13C-3d) 13
CC7H9N2O MW: 148.17 g.mol-1 Yield: 91% White solid
The [13C] 3,4-dihydroquinazolin-2(1H)-one 13C-3d was prepared accordingly to the general procedure, using the 2(azidomethyl)aniline (14.8 mg, 0.10 mmol), PPhMe2 (17.0 μL, 0.10 mmol) and 13CO2 (0.15 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from CH2Cl2 100% to CH2Cl2/MeOH 95/5) afforded the [13C] 3,4dihydroquinazolin-2(1H)-one 13C-3d as a white solid (13.3 mg, 91%). 1
H NMR (400 MHz, CD3OD) δ 7.14 (t, J = 7.7 Hz, 1H), 7.06 (dd, J = 7.6, 0.8 Hz, 1H), 6.93 (td, J = 7.5, 1.1 Hz, 1H), 6.78 (dd, J = 8.0, 0.7 Hz, 1H), 4.45 (d, J = 3.6 Hz, 2H). 13 C NMR (100 MHz, CD3OD) δ 157.9, 138.4, 129.0, 126.8, 123.3, 119.1, 115.1 (d, J = 3.3 Hz), 44.1. IR (cm-1) 3198, 3032, 1676, 1609, 1482, 1437, 1300, 1263, 1203, 736, 720, 618. HRMS (ESI) m/z calcd for 13CC7H9N2O [M+H]+ : 150.0743; found: 150.0741.
[13C] 3,7-dimethyl-1-(5-(2-oxotetrahydropyrimidin-1(2H)-yl)hexyl)-1H-purine-2,6(3H,7H)-dione (13C-3e) C1613CH26N6O3 MW: 363.42 g.mol-1 Yield: 81% Colorless oil The [13C]3,7-dimethyl-1-(5-(2-oxotetrahydropyrimidin-1(2H)-yl)hexyl)-1H-purine-2,6(3H,7H)-dione 13C-3e was prepared accordingly to the general procedure, using 1-(5-((3-azidopropyl)amino)hexyl)-3,7-dimethyl-1H-purine-2,6(3H,7H)-dione (31.2 mg, 0.086 mmol), PPhMe2 (12.2μL, 0.086 mmol) and 13CO2 (0.086 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (DCM : MeOH 95/5) afforded the [13C]3,7-dimethyl-1-(5-(2-oxotetrahydropyrimidin-1(2H)yl)hexyl)-1H-purine-2,6(3H,7H)-dione 13C-3e as a colorless oil (25.3 mg, 81%). 1
H NMR (400 MHz, CD3OD) δ 7.86 (d, J = 0.5 Hz, 1H), 4.43 (ddd, J = 9.5, 6.4, 3.0 Hz, 1H), 4.00 – 3.94 (m, 5H), 3.51 (s, 3H), 3.24 – 3.10 (m, 4H), 1.87 (p, J = 5.9 Hz, 2H), 1.68-1.53 (m, 3H), 1.51 – 1.39 (m, 1H), 1.35 – 1.24 (m, 3H), 1.09 (d, J = 6.8 Hz, 3H). 13 C NMR (100 MHz, CD3OD) δ 158.60, 156.5, 153.0, 149.9, 144.0, 108.8, 42.0, 40.8, 39.3, 34.2, 34.0, 30.1, 28.8, 24.9, 23.1, 23.0, 18.3. IR (cm-1) 2935, 1703, 1658, 1550, 1492, 1441, 1359, 1310, 1233, 763. LCMS (ESI) m/z C16H14N4O2 [M+H]+ 264.5.
[13C] (R)-4-phenylimidazolidin-2-one (13C-3f) S35
C813CH10N2O MW: 162.19 g.mol-1 Yield: 54% White solid
The [13C] (R)-4-phenylimidazolidin-2-one 13C-3f was prepared accordingly to the general procedure, using (R)-2-azido-1phenylethan-1-amine (16.2 mg, 0.10 mmol), PPhMe2 (17 μL, 0.10 mmol) and 13CO2 (0.150 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from CH2Cl2 100% to CH2Cl2/MeOH 95/5) afforded the [13C] (R)-4phenylimidazolidin-2-one 13C-3f as a white solid (8.7 mg, 54%). 1
H NMR (400 MHz, CD3OD) δ 7.41 – 7.35 (m, 4H), 7.32 – 7.27 (m, 1H), 4.89 (m, 1H, under water signal), 3.87 (td, J = 9.1, 3.7 Hz, 1H), 3.26 (ddd, J = 9.0, 7.3, 2.6 Hz, 1H). 13 C NMR (100 MHz, CD3OD) δ 166.7, 143.7, 129.8 (2C), 128.9, 127.1 (2C), 57.80 (d, J = 3.9 Hz), 50.67 (d, J = 3.2 Hz). IR (cm-1) 3173, 3030, 2864, 1651, 1487, 1451, 1254, 754, 698, 602, 495. TLC Rf: (DCM/MeOH, 95:5) = 0.18. HRMS (ESI) m/z calcd for 13CC8H11N2O [M+H]+ : 164.08994; found: 164.0898. [13C] 2,3-dihydro-1H-benzo[d]imidazo[1,5-a]imidazol-1-one (13C-3g) C813CH7N3O MW: 174.16 g.mol-1 Yield: 24% White solid The [13C] 2,3-dihydro-1H-benzo[d]imidazo[1,5-a]imidazol-1-one 13C-3g was prepared accordingly to the general procedure, using 2-(azidomethyl)-1H-benzo[d]imidazole (25.3 mg, 0.15 mmol), PPhMe2 (20.0 μL, 0.150 mmol) and 13CO2 (0.150 mmol) for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from cHex 50% to 100% EtOAc) afforded the [13C] 2,3-dihydro-1H-benzo[d]imidazo[1,5-a]imidazol-1-one 13C-3g as a white solid (3 mg, 24%). 1
H NMR (400 MHz, MeOD) δ 7.79 – 7.75 (m, 1H), 7.62-7.59 (m, 1H), 7.29 (dd, J = 6.1, 3.2 Hz, 2H), 4.54 (d, J = 2.9 Hz, 2H). C NMR (100 MHz, MeOD) δ 159.7, 153.3, 125.4, 125.1, 120.5 (2C), 113.0 (2C), 42.4. IR (cm-1) 3232, 2922, 1702, 1620, 1561, 1482, 1444, 14031325, 1272, 1176, 759, 744, 722, 668, 582. LCMS (ESI) m/z C813CH7N3O [M+H]+ 175.2. 13
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II. 6. Synthesis of [13C]Drug derivatives [13C] Oxatomide (13C-4a) C2613CH30N4O MW: 427.55 g.mol-1 Yield: 82% Beige solid
The [13C] Oxatomide 13C-4a was prepared accordingly to the general procedure, using 2-azido-N-(3-(4benzhydrylpiperazin-1-yl)propyl)aniline (42.6 mg, 0.10 mmol), PPhMe2 (14.2 μL, 0.110 mmol) and 13CO2 (0.110 mmol) in 0.50 mL of DMF for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 1/9 to DCM : MeOH 97/3) afforded the [13C] Oxatomide 13C-4a as a beige solid (35 mg, 82%). 1
H NMR (400 MHz, DMSO-d6) δ 10.81 (d, J = 4.7 Hz, 1H), 7.41 (d, J = 7.2 Hz, 4H), 7.28 (t, J = 7.6 Hz, 4H), 7.17 (t, J = 7.3 Hz, 2H), 7.13 – 7.08 (m, 1H), 6.98-6.94 (m, 3H), 4.25 (s, 1H), 3.78 (m, 2H), 2.32 (br. m, 10H), 1.77 (br. m, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 154.5, 154.1, 152.3, 142.8, 130.3 (d, J = 6.4 Hz), 128.5 (4C), 128.2 (d, J = 4.2 Hz), 127.5 (4C), 126.8, 120.6, 120.4, 108.6 (d, J = 3.8 Hz), 107.6 (d, J = 3.4 Hz), 87.3, 75.5 (2C), 54.4 (2C), 52.6, 51.2, 37.9 . IR (cm-1) 2926, 2811, 1650, 1489, 1154, 740, 705. TLC Rf: (CH2Cl2/EtOAc, 97:3) = 0.45. HRMS (ESI) m/z calcd for C2613CH30N4O [M+H]+: 428.2525, found 428.2626. LCMS (ESI) m/z C2613CH30N4O [M+H]+ 428.7. [13C] Domperidone (13C-4b) C2113CH24ClN5O2 MW: 426.9 g.mol-1 Yield: 75% Beige solid
The [13C] Domperidone 13C-4b was prepared accordingly to the general procedure, using 1-(1-(3-((2azidophenyl)amino)propyl)piperidin-4-yl)-5-chloro-1,3-dihydro-2H-benzo[d]imidazol-2-one (16.0 mg, 0.030 mmol), PPhMe2 (5.30 μL, 0.030 mmol) and 13CO2 (0.110 mmol) in 0.45 mL of DMF-d7 for 10 minutes. Purification by Flash Chromatography on SiO2 gel (DCM : MeOH 97/3) afforded the [13C]Domperidone 13C-4b as a beige solid (12 mg, 75%). S37
1
H NMR (400 MHz, CD3OD) δ 7.37 – 7.20 (m, 3H), 7.13 – 7.05 (m, 4H), 4.26 (ddt, J = 15.8, 12.0, 3.8 Hz, 1H), 4.00 (td, J = 6.7, 4.1 Hz, 2H), 3.08 (br. d, J = 11.8 Hz, 2H), 2.53 – 2.49 (m, 2H), 2.40 (qd, J = 12.6, 3.8 Hz, 2H), 2.20 – 2.09 (m, 2H), 2.05 – 1.98 (m, 2H), 1.74 – 1.71 (br. m, 2H), 1.29 (br. s, 2H). 13 C NMR (100 MHz, CD3OD) δ 156.8, 156.2, 131.7, 131.6, 130.7, 129.8, 129.1, 128.3, 127.9, 122.7, 122.5, 122.0, 111.6, 110.6, 110.5, 109.3 (d), 56.5, 54.2, 51.9, 39.9, 29.5, 26.6. IR (cm-1) 3178, 2928, 1693, 1649, 1487, 1373, 742, 708, 593. TLC Rf: (CH2Cl2/MeOH, 97:3) = 0.55. LCMS (ESI) m/z C2113CH24ClN5O2 [M+H]+ 427.5. HRMS (ESI) m/z calcd for C2113CH24ClN5O2 [M+H]+ 428.252672; found: 428.252593.
[13C] NR2-receptor antagonist (13C-4c) C2113CH27ClN4O MW: 357.14 g.mol-1 Yield: 86% White solid
The [13C] NR2-receptor antagonist 13C-4c was prepared accordingly to the general procedure, using 2-azido-N-(5-(4-(4chlorophenyl)piperazin-1-yl)pentyl)aniline (35.2 mg, 0.10 mmol), PPhMe2 (14.2 μL, 0.110 mmol) and 13CO2 (0.110 mmol) in 0.50 mL of DMF for 10 minutes. Crystallisation from DCM/Pentane afforded the [13C] NR2-receptor antagonist 13C-4c as a white solid (30.7 mg, 86%). 1
H NMR (400 MHz, CDCl3) δ 9.42 (d, J = 3.8 Hz, 1H), 7.19 (d, J = 9.0 Hz, 2H), 7.12 – 7.04 (m, 3H), 7.00 (d, J = 7.6 Hz, 1H), 6.81 (d, J = 9.1 Hz, 2H), 3.90 (td, J = 7.2, 4.0 Hz, 2H), 3.18 (m, 4H), 2.62 (m, 4H), 2.42 (s, 2H), 1.82 (qt, J = 7.4 Hz, 2H), 1.62 (s, 2H), 1.43 (qt, J = 7.4 Hz, 2H). 13 C NMR (100 MHz, CDCl3) δ 155.8, 155.4, 149.9, 130.5 (d, J = 6.5 Hz), 129.1 (2C), 127.9 (d, J = 4.4 Hz), 124.8, 121.5 (d, J = 10.2 Hz), 117.4 (2C), 109.6 (d, J = 3.9 Hz), 108.0 (d, J = 3.5 Hz), 58.4, 53.1 (2C), 49.0 (2C), 40.8, 28.3, 26.3, 24.8. IR (cm-1) 3415, 2936, 1818, 2120, 1600, 1517, 1496, 1446, 1288, 1235, 1148, 816, 738 TLC Rf: (CH2Cl2/EtOAc 97:3) = 0.40. HRMS (ESI) m/z calcd for 13CC21H28ClN4O [M+H]+ : 400.1979; found: 400.1978.
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[13C] Flibanserin (13C-4d) C1913CH21F3N4O MW: 391.39 g.mol-1 Yield: 84% Colorless oil
The [13C] Flibanserin 13C-4d was prepared accordingly to the general procedure, using 2-azido-N-(2-(4-(3(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)aniline (18.5 mg, 0.048 mmol), PPhMe2 (6.80 μL, 0.048 mmol) and 13CO2 (0.050 mmol) in 0.50 mL of DMF-d7 for 10 minutes. Purification by Flash Chromatography on SiO2 gel (from cHex : EtOAc 3/7 RF: 0.25) afforded the [13C] Flibanserin 13C-4d as a colorless oil (15.6 mg, 84%). 1
H NMR (400 MHz, CD3OD) δ 7.27 (t, J = 7.9 Hz, 1H), 7.08 – 6.94 (m, 7H), 3.97 (ddd, J = 10.6, 7.1, 4.0 Hz, 2H), 3.12 – 3.10 (m, 4H), 2.67 (t, J = 6.7 Hz, 2H), 2.65-2.61 (m, 4H), 1.18 (br. m, 1H). 13 C NMR (100 MHz, CD3OD) δ 156.8, 153.0, 132.5, 132.2, 131.6, 131.5, 130.8, 129.6, 127.2, 124.5, 122.7, 122.4, 120.2, 116.6, 112.8, 110.5, 109.3, 56.7, 54.1, 39.0. IR (cm-1) 2944, 2826, 1651, 1490, 1449, 1318, 1162, 1120, 742. TLC Rf: (cHex/EtOAc, 3:7) = 0.25. LCMS (ESI) m/z C1913CH21F3N4O [M+H]+: 392.6. HRMS (ESI) m/z calcd for C1913CH21F3N4O 6 [M+H]+ 392.1773; found: 392.1771.
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II. 7. Experimental procedure for Carbon-14 radiolabeling General procedure for Carbon-14 labeling 14
CO2 (2.17 GBq mmol-1) was generated using a 14CO2 manifold system (RC Tritec AG). Mass spectra (ESI) for the calculation of specific activities were obtained using a Waters Micromass ZQ spectrometer. Radiochemical purities were determined by Thin Layer Chromatography on TLC silica gel 60F254 glass plates (Merck) using a RITA scanner (Raytest) for the radioactive detection
[14C] 1H-benzo[d]imidazol-2(3H)-one (14C-2a)
C7H6N2O (unlabeled compound) Radioactive yield: 95% white solid
In a NMR tube with a J Young valve were added a solution of 2-azidoaniline (16 mg, 0.119 mmol) in CD3CN (0.58 mL), diphenylmethane (19.9 µL, 0.119 mmol) and PPhMe2 (17 µL, 0.119 mmol). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (1.85 GBq mmol-1, 203.50 MBq, 0.110 mmol) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold system and kept at room temperature for 5 minutes. During this time, a precipitate formed. After concentration, the crude residue was purified on silica gel using gradient-flash chromatography (50% ethyl acetate in heptane to 100% ethyl acetate). [2a-14C]-1H-Benzo[d]imidazol-2(3H)-one was obtained as a white solid (193.51 MBq, 0.105 mmol, radioactive yield: 95%) 1
H NMR (400MHz, CD3OD) δ 7.1-7.0 (m, 4H). In agreement with an authentic sample. MS (ESI) m/z C7H7N2O [M+H]+ 135 (12C isomer), 137 (14C isomer). Specific activity: 1.85 GBq mmol-1. GC/MS (EI) m/z C7H6N2O [M]+ 134 (24%, 12C isomer), 136 (100%, 14C isomer). TLC (silicagel 60F254, CH2Cl2/MeOH (100/5)) Rf=0.31. Radioactive purity: 99.5%. Co-elution with an authentic sample.
[14C] Oxatomide (14C-4a) C2614CH30N4O2 MW: 428.56 g.mol-1 Yield: 66% Beige solid
In a NMR tube with a J Young valve were added a solution of azido precursor (42.6 mg, 0.1 mmol, 1 eq.) in CH3CN (0.5 mL) and PPhMe2 (14.2 µL, 0.1 mmol, 1 eq.). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (2.17 GBq mmol-1, 303.8 MBq, 0.14 mmol, 1.4 eq.) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold
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system and kept at RT for 10 minutes. After concentration, purification using Flash Chromatography on SiO2 (CH2Cl2/MeOH (100/5)) gave [14C]-oxatomide 14C-4a (2.13 GBq mmol-1, 141.34 MBq, 0.066 mmol, yield: 66%). Specific activity (MS (ESI)): 2.13 GBq mmol-1. TLC (silicagel 60F254, CH2Cl2/MeOH (100/5)) Rf=0.41. Radiochemical purity: ≥99%. [14C] Domperidone (14C-4b) C2114CH24ClN5O2 MW: 427.91 g.mol-1 Yield: 57% Beige solid
In a NMR tube with a J Young valve were added a solution of azido precursor (16 mg, 0.037 mmol, 1 eq.) in DMF (0.5 mL) and PPhMe2 (5.3 µL, 0.037 mmol, 1 eq.). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (2.17 GBq mmol-1, 171.43 MBq, 0.079 mmol, 2.1 eq.) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold system and kept at RT for 10 minutes. After concentration, purification using Flash Chromatography on SiO2 (CH2Cl2/MeOH (100/10)) gave [14C]-domperidone 14C-4b (2.08 GBq mmol-1, 41.81 MBq, 0.02 mmol, yield: 54%) Specific activity (MS (ESI)): 2.08 GBq mmol-1. TLC (silicagel 60F254, CH2Cl2/MeOH (100/10)) Rf=0.31. Radiochemical purity: ≥98%. [14C] NR2-receptor antagonist (14C-4c) C2114CH27ClN4O MW: 400.93 g.mol-1 Yield: 89% White solid
In a NMR tube with a J Young valve were added a solution of azido precursor (18.7 mg, 0.047 mmol, 1 eq.) in CH 3CN (0.5 mL) and PPhMe2 (6.6 µL, 0.05 mmol, 1 eq.). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (2.17 GBq mmol-1, 195.47 MBq, 0.09 mmol) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold system and kept at RT for 10 minutes. After concentration, purification using Flash Chromatography on SiO2 (80% ethyl acetate in heptane to 100% ethyl acetate) gave [14C]-5HT7 14C-4c (2.09 GBq mmol-1, 86.95 MBq, 0.042 mmol, yield: 89%). Specific activity (MS (ESI)): 2.09 GBq mmol-1. TLC (silicagel 60F254, CH2Cl2/MeOH (100/5)) Rf=0.29. Radiochemical purity: ≥99%.
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[14C] Flibanserin (14C-4d) C1914CH21F3N4O MW: 392.40 g.mol-1 Yield: 95%
In a NMR tube with a J Young valve were added a solution of azido precursor (9 mg, 0.023 mmol, 1 eq.) in DMF-d7 (0.4 mL) and PPhMe2 (3.3 µL, 0.023 mmol, 1 eq.). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (2.17 GBq mmol-1, 84.63 MBq, 0.039 mmol, 1.7 eq.) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold system and kept at RT for 10 minutes. After concentration, purification using Flash Chromatography on SiO2 (heptane /AcOEt (20/80)) gave [14C]-flibanserin 14C-4d (2.04 GBq mmol-1, 44.03 MBq, 0.022 mmol, yield: 95%). Specific activity (MS (ESI)): 2.04 GBq mmol-1. TLC (silicagel 60F254, Heptane/AcOEt (20/80)) Rf=0.33. Radiochemical purity: 99%. [14C]-SDWSTEK (14C-5) C3914CH57N11O17 MW: 965.95 g.mol-1 Yield: 43%
In a NMR tube with a J Young valve were added a solution of azido precursor S34 (6.8 mg, 0.007 mmol, 1 eq.) in DMF-d7 (0.5 mL), trimethylamine (5 µL, 0.035 mmol, 5 eq.) and PPhMe 2 (1 µL, 0.007 mmol, 1 eq.). The NMR tube was attached to a 14C manifold system and the reaction mixture was frozen with a liquid nitrogen bath. The NMR tube was vacuumized and 14CO2 (2.17 GBq mmol-1, 203.98 MBq, 0.094 mmol, 13.4 eq.) was condensed using the freezing bath. The NMR tube was closed, detached from the manifold system and kept at room temperature for 10 minutes. After concentration, purification by preparative HPLC gave [14C]-SDWSTEK –(14C-5) (2.09 GBq mmol-1, 5.81 MBq, 0.003 mmol, yield: 43%) . Preparative HPLC conditions: Column: HyPURITY C18 5µ (250 x 10 mm) from Thermo Flow: 5 mL/min. Mobile phase: CH3CN/H2O/HCOOH (5/95/0.1) to CH3CN/H2O/HCOOH (15/85/0.1) - 24 min S42
CH3CN/H2O/HCOOH (15/85/0.1) – 60 min Specific activity (MS (ESI)): 2.09 GBq mmol-1.
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II. 8. Experimental procedure for Carbon-11 radiolabeling. General procedure performed at Service Hospitalier Frédéric Joliot (Orsay, France): Automated radiosynthesis with carbon-11 was performed using a MeIplus research synthesizer (Synthra GmbH, Germany) with modifications to undergo direct bubbling of [11C]CO2 into the reaction vessel (figure 2, see supporting information). No carrier-added [11C]CO2 (3.5-18 GBq) was produced via the 14N(p, α)11C nuclear reaction by irradiation of a [14N]N2 target containing 0.15-0.5% of O2 on a cyclone 18/9 cyclotron (18 MeV, IBA, Belgium) and trapped at -180 °C. [11C]CO2 was then released at 50 °C under a stream of helium (8 mL/min) to bubble for 10 s into the reaction vessel containing a solution of the precursor (1 mg) and dimethylphenyl phosphine (15 µL) in DMF (300 µL) at -50 °C. The mixture was heated at 20 °C for 5 min and hydrolyzed with glacial acetic acid (100 µL) followed by a mixture of CH 3CN/H2O (1 mL, 50/50 v/v). Quality control is performed by HPLC using a 717plus Autosampler system equipped with a 1525 binary pump and a 2996 photodiode array detector (Waters, USA) and a Flowstar LB 513 (Berthold, France) gamma detector. The system was monitored with the Empower 3 (Waters, USA) software. HPLC were realized on a reverse phase analytical Symmetry C18 (150 x 3.9 mm, 5 μm, Waters, USA) column using a mixture of H2O/CH3CN/PicB7® (proportions depending on the compound, 2 mL/min) as eluent. UV detection was performed at the maximum absorbance of the compound. Identification of the peak was assessed by comparing the retention time of carbon-11 labeled compound with the retention time of the non-radioactive reference (tRref). For acceptance, the retention time must be within the tRref ± 10% range. Radiochemical purity (RCP) was calculated as the ratio of the area under the curve (AUC) of the peak over the sum of the AUCs of all other peaks on gamma chromatograms. Radiochemical purity is the mean values of three consecutive runs. The radiochemical yield (RCY) of the labeling reaction was calculated as the ratio of the decaycorrected activity and the end of the synthesis (AEOS), measured in an ionization chamber (Capintec®, Berthold, France) over the starting activity of [11C]CO2 (ACO2) measured by the calibrated detector of the synthesizer. This ratio was corrected for the radiochemical purity following the equation: RCY = (AEOS / ACO2) x RCP. Molar activity was calculated as the ratio of the activity of the collected peak of the radioactive product measured in an ionization chamber (Capintec®, Berthold, France) over the molar quantity of the compound determined using calibration curves. Molar activity was calculated as the mean value of three consecutive runs.
figure 2. Modified MeIplus Research module for direct CO2 labeling.
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[11C] 1H-benzo[d]imidazol-2(3H)-one (11C-2a) C611CH6N2O RCYield: 79%
Compound 11C-2a (2.4 GBq) was synthesized from compound 1a according to the general procedure within 15 minutes in 79% RCY and 100% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 90/10/0.2 v/v/v, 2 mL/min, λ = 279 nm). [13C] 5-chloro-1H-benzo[d]imidazol-2(3H)-one (11C-2b) C611CH5ClN2O RCYield: 45%
Compound 11C-2b (1.4 GBq) was synthesized from compound S1 according to the general procedure within 15 minutes in 45% RCY and 98% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 80/20/0.2 v/v/v, 2 mL/min, λ = 287 nm). [11C] 5-fluoro-1H-benzo[d]imidazol-2(3H)-one (11C-2c) C611CH5FN2O RCYield: 43%
Compound 11C-2c (1.9 GBq) was synthesized from compound S2 according to the general procedure within 15 minutes in 43% RCY and 73% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 85/15/0.2 v/v/v, 2 mL/min, λ = 284 nm). [11C] 5-methoxy-1H-benzo[d]imidazol-2(3H)-one (11C-2e) C711CH8N2O2 RCYield: 70%
Compound 11C-2e (2.6 GBq) was synthesized from compound S4 according to the general procedure within 15 minutes in 70% RCY and 99% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 90/10/0.2 v/v/v, 2 mL/min, λ = 290 nm).
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[11C] tert-butyl (2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate (11C-2f) C1111CH15N3O3 RCYield: 65 %
Compound 11C-2f (2.1 GBq) was synthesized from compound S5 according to the general procedure within 15 minutes in 65% RCY and 99% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 80/20/0.2 v/v/v, 2 mL/min, λ = 290 nm).
[11C] methyl 2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-carboxylate (11C-2g) C811CH8N2O3 RCYield: 58%
Compound 11C-2g (2.3 GBq) was synthesized from compound S6 according to the general procedure within 15 minutes in 58% RCY and 97% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 90/10/0.2 v/v/v, 2 mL/min, λ = 266 nm). [11C] 5-(trifluoromethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2h) C711CH5F3N2O RCYield: 28%
Compound 11C-2h (0.8 GBq) was synthesized from compound S7 according to the general procedure within 15 minutes in 28% RCY and 94% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 75/25/0.2 v/v/v, 2 mL/min, λ = 282 nm). [11C] 1H-naphtho[1,2-d]imidazol-2(3H)-one (11C-2i) C1011CH8N2O3 RCYield: 76%
Compound 11C-2i (2.7 GBq) was synthesized from compound S8 according to the general procedure within 15 minutes in 76% RCY and 99% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 80/20/0.2 v/v/v, 2 mL/min, λ = 241 nm).
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[11C] 4,6-dimethyl-1H-benzo[d]imidazol-2(3H)-one (11C-2j) C611CH10N2O RCYield: 79%
Compound 11C-2j (2.7 GBq) was synthesized from compound S9 according to the general procedure within 15 minutes in 76% RCY and 99% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 80/20/0.2 v/v/v, 2 mL/min, λ = 286 nm).
[13C] 4-phenyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (11C-2k) C1211CH10N2O RCYield: 66%
Compound 11C-2k (2.0 GBq) was synthesized from compound S10 according to the general procedure within 15 minutes in 66% RCY and 96% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 70/30/0.2 v/v/v, 2 mL/min, λ = 256 nm).
[11C] 1-benzyl-1H-benzo[d]imidazol-2(3H)-one (11C-2l) C1311CH12N2O RCYield: 55 %
Compound 11C-2l (2.1 GBq) was synthesized from compound S11 according to the general procedure within 15 minutes in 55% RCY and 100% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 70/30/0.2 v/v/v, 2 mL/min, λ = 240 nm).
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[11C] 1-(naphthalen-2-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2m) C1711CH14N2O RCYield: 56%
Compound 11C-2m (1.9 GBq) was synthesized from compound S12 according to the general procedure within 15 minutes in 56% RCY and 96% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 60/40/0.2 v/v/v, 2 mL/min, λ = 277 nm).
[11C] 1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2n) C1411CH12N2O3 RCYield: 62%
Compound 11C-2n (2.4 GBq) was synthesized from compound S13 according to the general procedure within 15 minutes in 62% RCY and 96% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 65/35/0.2 v/v/v, 2 mL/min, λ = 280 nm).
[11C] 1-benzyltetrahydropyrimidin-2(1H)-one (11C-3a) C1011CH14N2O RCYield: 68%
Compound 11C-3a (2.1 GBq) was synthesized from compound X according to the general procedure within 15 minutes in 68% RCY and 100% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 65/35/0.2 v/v/v, 2 mL/min, λ = 220 nm).
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[11C] 7-methyl-1-(5-(2-oxotetrahydropyrimidin-1(2H)-yl)hexyl)-3,4,5,7-tetrahydro-1H-purine-2,6-dione (11C-3e ) C1611CH26N6O3 RCYield: 90%
Compound 11C-3e (2.7 GBq) was synthesized from compound S19 according to the general procedure within 15 minutes in 90% RCY and 100% RCP as calculated after analysis by HPLC (H2O/CH3CN/PicB7® 85/15/0.2 v/v/v, 2 mL/min, λ = 280 nm). [11C] Oxatomide (11C-4a) C2611CH30N4O RCYield: 45%
The crude product was synthesized from compound S30 following the general procedure. Purification by semipreparative HPLC was performed on a reverse phase Symmetry C18 column (250 x 4.6 mm, 5 μm, Agilent, USA) using a mixture of H2O/CH3CN/TFA (55/45/0.1 v/v/v, 5 mL/min) as eluent with gamma and UV (λ = 280 nm) detection. The collected peak (tR = 4.8-5.5 min) of 11C-oxatomide was diluted with water (20.0 mL) and loaded on a C18 cartridge (SepPak C18, Waters, USA). The cartridge was rinsed with water (10.0 mL) and the product was eluted with ethanol (2.0 mL) and further diluted with aq. 0.9 % NaCl (18.0 mL). Ready-to-inject 11C-oxatomide 11C-4a (3.6 ± 0.2 GBq) was obtained within 30 minutes from end of beam in 45 ± 2% RCY and 70 ± 5 GBq/µmol molar activity (n = 2). Quality control was performed following the general procedure (H2O/CH3CN/PicB7® 65/35/0.2 v/v/v, 2 mL/min, λ = 280 nm).
[11C] Domperidone (11C-4b) C2111CH24ClN5O2 RCYield: 43%
The crude product was synthesized from compound S33 following the general procedure. Purification by semipreparative HPLC was performed on a reverse phase Symmetry C18 column (250 x 4.6 mm, 5 μm, Agilent, USA) using a S49
mixture of H2O/CH3CN/TFA (65/35/0.1 v/v/v, 5 mL/min) as eluent with gamma and UV (λ = 280 nm) detection. The collected peak (tR = 5.5-6.3 minutes) of 11C-domperidone was diluted with water (20.0 mL) and loaded on a C18 cartridge (Sep-Pak C18, Waters, USA). The cartridge was rinsed with water (10.0 mL) and the product was eluted with ethanol (2.0 mL) and further diluted with aq. 0.9 % NaCl (18.0 mL). Ready-to-inject 11C-domperidone 11C-4b (5.2 ± 0.4 GBq) was obtained within 30 minutes from end of beam in 43 ± 5% RCY and 74 ± 20 GBq/µmol molar activity (n = 3). Quality control was performed following the general procedure (H2O/CH3CN/PicB7® 70/30/0.2 v/v/v, 2 mL/min, λ = 285 nm).
[11C] 5HT7-receptor antagonist (11C-4c) C2111CH27ClN4O RCYield: 39%
The crude product was synthesized from compound S28 following the general procedure. Purification by semipreparative HPLC was performed on a reverse phase Symmetry C18 column (250 x 4.6 mm, 5 μm, Agilent, USA) using a mixture of H2O/CH3CN/TFA (65/35/0.1 v/v/v, 5 mL/min) as eluent with gamma and UV (λ = 280 nm) detection. The collected peak (tR = 9.0-10.0 min) of 11C-4c was diluted with water (20.0 mL) and loaded on a C18 cartridge (Sep-Pak C18, Waters, USA). The cartridge was rinsed with water (10.0 mL) and the product was eluted with ethanol (2.0 mL) and further diluted with aq. 0.9 % NaCl (18.0 mL). Ready-to-inject 11C-4c (4.2 ± 0.3 GBq) was obtained within 30 min from end of beam in 39 ± 3% RCY and 70 ± 7 GBq/µmol molar activity (n = 2). Quality control was performed following the general procedure (H2O/CH3CN/PicB7® 65/35/0.2 v/v/v, 2 mL/min, λ = 252 nm).
[11C] Flibanserin (11C-4d) C2111CH27ClN4O RCYield: 39%
The crude product was synthesized from compound S32 following the general procedure. Purification by semipreparative HPLC was performed on a reverse phase Symmetry C18 column (250 x 4.6 mm, 5 μm, Agilent, USA) using a mixture of H2O/CH3CN/TFA (55/45/0.1 v/v/v, 5 mL/min) as eluent with gamma and UV (λ = 280 nm) detection. The collected peak (tR = 2.8-3.5 min) of 11C-flibanserin 11C-4d was diluted with water (20 mL) and loaded on a C18 cartridge (Sep-Pak C18, Waters, USA). The cartridge was rinsed with water (10 mL) and the product was eluted with ethanol (2 mL) and further diluted with aq. 0.9 % NaCl (18 mL). Ready-to-inject 11C-flibanserin 11C-4d (3.7 ± 0.4 GBq) was obtained within 30 min from end of beam in 48 ± 2% RCY and 75 ± 7 GBq/µmol molar activity (n = 2). Quality control was performed following the general procedure (H2O/CH3CN/PicB7® 55/45/0.2 v/v/v, 2 mL/min, λ = 280 nm). S50
[14C]-SDWSTEK (11C-5) C3911CH57N11O17 RCYield: 23%
Compound [11C]-5 (0.9 ± 0.2 GBq) was synthesized from compound S34 and triethylamine (10.0 µL) according to the general procedure within 15 minutes in 23% RCY and 100% RCP as calculated after analysis by HPLC (H2O/CH3CN/TFA gradient, 2 mL/min, λ = 282 nm). Note: In order to verify that [11C]CO2 was labeled on the azide-modified amino-acid of the peptide and not on the lysine moiety, the same protocol was carried out in the absence of the phosphine. In those conditions, no labeling was observed and the totality on the initial amount of [11C]CO2 was recovered on an ascarite® trap after hydrolysis with acetic acid. This experiment demonstrates that the presence of the phosphine is mandatory for the labeling of the peptide with [11C]CO2
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II. 9. Experimental procedure for Carbon-11 radiolabeling (Karolinska Institutet, Stockholm, Sweden) Radiochemistry All reactions were carried out in oven-dried, septum-equipped glass vials under inert gas. No-carrier-added [11C]CO2 was produced using 16.5 MeV protons in the 14N(p,α)11C nuclear reaction on a mixture of nitrogen and oxygen gas (1% oxygen). After 1 minute irradiation at 10 µA, typically 750 MBq of [11C]CO2 was obtained. A [11C]CO-synthesizer prototype from GEMS PET Systems AB (GE Healthcare, Uppsala, Sweden) was used as a synthetic platform for this study. At the end of bombardment (EOB), the target content was delivered to the [11C]CO-synthesizer prototype, where the [11C]CO2 was trapped on a molecular sieve column (0.60 g packed in a ¼” SS tube, 4 Å, mesh 80/100, GRACE) at room temperature (RT). The accumulated [11C]CO2 was released into a controlled stream (10 ml/min, Mass flow controller, Bronckhorst) of He (helium 6.0) while heating the molecular sieve column to 360°C. The purified [11C]CO2 was further concentrated on a silica gel (10 mg, 60 Å, 60-100 mesh) trap immersed in liquid nitrogen. General procedure for radiosynthesis The appropriate azide (1mg) was charged into a V-shaped reaction vessel (4.50 mL, Supelco) which was subsequently purged with N2 for 10-20 min. Next, DMF (0.45 ml) was added and the mixture was purged for an additional 5 minutes. Right before the end of bombardment, phosphine (15 μL) was added and the vial was submerged into a dry ice/EtOH cooling bath (approx. -50 °C). [11C]CO2 (handled as described above) was bubbled into the reaction vessel in a stream of He (at 5 mL/min) until maximum radioactivity was observed in the vessel. The outlet of the vial was equipped with an Ascarite trap to measure untrapped [11C]CO2. Following completed entrapment, the needles were removed and the vial was heated at 90 °C for 3 minutes and allowed to cool at room temperature for 2 minutes. The mixture was finally quenched by addition of conc. acetic acid and purged with N2 to trap the unreactive [11C]CO2 in the Ascarite. Analysis and radiochemical yield Decay-corrected radiochemical yields (RCYs) were determined by radio-HPLC of the crude reaction mixture using an Agilent 1220 Infinity LC system with a built-in photodiode array (PDA) UV-detector (254 nm was used), in series with a Eckert & Ziegler β+-flow detector. An Agilent Eclipse XDB-C18 reverse-phase column (4.6 x 150 mm, 5 µm) was used with a flow rate of 2.5 mL /min, and a gradient of acetonitrile and ammonium formate (MeCN / HCOONH 4 (0.1 M) 10:90 to 90:10 in 5 minutes) was used as mobile phase. Corresponding references were used to identify the 11C-labelled compounds. Semi-preparative-HPLC purification Semi-preparative high performance liquid chromatographic (HPLC) was performed using a TRACERlab FX C Pro chemistry synthesizer (GE, Uppsala, Sweden) with a built-in β+-flow detector in series with a UV absorbance detector. An Atlantis Prep T3 reverse phase column (10 x 150 mm, 5 µm) was eluted with a mixture of acetonitrile and ammonium formate (MeCN / HCOONH4 (0.1 M), 18:82) at 5 mL/min. The fraction containing the target compound was collected and measured in a dose calibrator for calculation of the isolated radiochemical yield. Calculation of the molar activity Molar activity (GBq/µmol) was calculated by dividing the radioactivity (GBq) with the chemical amount of the tracer molecule (µmol). Analytical HPLC was used for determination of the concentration of carrier in the product solution. S52
Chromatographic separation was achieved on an Atlantis Prep T3 column eluted with MeCN- HCOONH4 (0.1 M) 14: 86 v/v at 4 mL / min.
[13C] 5-iodo-1H-benzo[d]imidazol-2(3H)-one (11C-2d) C611CH5IN2O RCYield: 85%
Compound 11C-2d (345 MBq) was synthesized from compound S4 according to the general procedure and keeping the vial for 2 minutes at 90 °C. Within 17 minutes the reaction yelded 85% RCY and 100% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
[11C] 1-(thiophen-2-ylmethyl)tetrahydropyrimidin-2(1H)-2-one (11C-3b) C811CH12N2OS RCYield: 95% Compound 11C-3b (250 MBq) was synthesized from compound S16 according to the general procedure and keeping the vial for 5 minutes at RT. Within 16 minutes the reaction yelded 95% RCY and 100% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
[11C]- [13C] 1-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)tetrahydropyrimidin-2(1H)-one (11C-3c) C1211CH15N3OS2 RCYield: 95 %
Compound 11C-3c (250 MBq) was synthesized from compound S17 according to the general procedure and keeping the vial for 5 minutes at RT °C. Within 17 minutes the reaction yelded 95% RCY and 100% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
S53
[11C]- 2,3-dihydro-1H-benzo[d]imidazo[1,5-a]imidazol-1-one (11C-3g) C1211CH15N3OS2 RCYield: 95 %
Compound 11C-3g (477 MBq) was synthesized from compound S18 according to the general procedure and keeping the vial for 5 minutes at RT °C. Within 17 minutes the reaction yelded 95% RCY and 100% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
[11C]-3,4-dihydroquinazolin-2(1H)-one (11C-3d) C711CH8N2O RCYield: 99 %
Compound 11C-3d (503 MBq) was synthesized from compound S25 according to the general procedure and keeping the vial for 5 minutes at RT. Within 23 minutes the reaction yielded 99% RCY and 99% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
[11C] (R)-4-phenylimidazolidin-2-one (13C-3f) C811CH10N2O RCYield: 99 %
Compound 11C-3f (648 MBq) was synthesized from compound S22 according to the general procedure and keeping the vial for 5 minutes at RT. Within 20 minutes the reaction yielded 99% RCY and 98% RCP as calculated after analysis by HPLC (MeCN / HCOONH4 (0.1 M) 10:90 to 90:10 in 5 min, λ = 254 nm).
S54
[11C] (S)-CGP-12177 (11C-4e) C1311CH21N3O3 RCYield: 34 %
The crude product was synthesized from compound 11C-4e following the general procedure. 11C-(S)-[11C]CGP-12177 was diluted with water and loaded on Atlantis Prep T3 reverse phase column (10 x 150 mm, 5 µm) with a mixture of acetonitrile and ammonium formate (MeCN / HCOONH4 (0.1 M), 18:82) at 5 mL/min) as eluent. The collected peak of 11 C-(S)-[11C]CGP-12177 was diluted with water and loaded on an Atlantis Prep T3 column eluted with MeCN- HCOONH4 (0.1 M) 14: 86 v/v at 4 mL / min. Ready-to-inject 11C-(S)-[11C]CGP-12177 11C-4e (34 ± 17 GBq) was obtained within 30 minutes from end of beam and 32.5 ± 16 GBq/µmol molar activity (n = 2).
S55
III. 1H and 13C NMR Data
S56
S57
S58
S59
S60
S61
S62
S63
S64
S65
S66
S67
S68
S69
S70
S71
S72
S73
S74
S75
S76
S77
S78
S79
S80
S81
S82
S83
S84
S85
S86
S87
S88
S89
S90
S91
S92
S93
S94
S95
S96
S97
S98
S99
S100
S101
S102
S103
S104
IV. Radio-HPLC Analysis for 11C-Labeled Compounds
S104
[11C]-3,4-dihydroquinazolin-2(1H)-one (11C-3d) Area % Report 12-37-52
5.023
Data File: C:\EZChrom Elite\Enterprise\Projects\Kaisa\Antonio\ADV-01-019-KH_2_11-15-2017 PM.dat Method: C:\EZChrom Elite\Enterprise\Projects\Default\Method\Kaisa\2017_02_14.met Acquired: 11/15/2017 12:39:05 PM Printed: 11/23/2017 12:32:05
0
1
2
3
3.637
3.170
2.200
3.033
Volts
2000
4
Volts
3000
Retention Time
1000
5
6
7
Minutes
Retention Time
20
20
2.267
2.700
mV
40
mV
40
0 0
1
2
3
4
5
6
7
Minutes
VWD: Signal A, 254 nm Results Retention Time
Area
Area %
Height
Height %
2.200 3.033 3.170 3.637 5.023
159359620 6683198 1627008 7380831 131127577
52.05 2.18 0.53 2.41 42.83
50838910 1233146 465768 2140375 41448289
52.89 1.28 0.48 2.23 43.12
S105
Totals
Agilent 35900E Interface Radio Results Retention Time 2.267
2.700
Area 23562869
129923
Area % 99.45
0.55
Height 4860555
25176
Height % 99.48
0.52
Totals 23692792
S106
100.00
4885731
100.00
[13C] (R)-4-phenylimidazolidin-2-one (13C-3f) Area % Report
Data File: Method: Acquired: Printed:
C:\EZChrom Elite\Enterprise\Projects\Kaisa\Antonio\ADV-01-020-KH_11-15-2017 C:\EZChrom Elite\Enterprise\Projects\Default\Method\Kaisa\2017_02_14.met 11/15/2017 4:01:48 PM 11/23/2017 12:34:20 PM
3-53-59 PM.dat
4000
0
1
2
3
4
5.620
5.030
3.540
3.020
2.270
2000
Volts
Volts
Retention Time
5
6
7
Minut
50
3.533
50
0
1
2
0 3
4
5
6
7
Minutes
VWD: Signal A, 254 nm Results Retention Time
Area
Area %
Height
Height %
2.270 3.020 3.540 5.030 5.620
8614357 6797063 6049453 249324223 2855974
3.15 2.48 2.21 91.11 1.04
2919332 1632450 1721879 64447478 939310
4.07 2.28 2.40 89.93 1.31
S107
mV
100
Retention Time
2.333
mV
100
Totals
Agilent 35900E Interface Radio Results Retention Time 2.333
Area 53038005
Area % 98.69
3.533
70340
1.31
Height 10408860
164322
Height %98.45
1.55
Totals 53741405
100.00
S108
10573182
100.00
[11C]- 2,3-dihydro-1H-benzo[d]imidazo[1,5-a]imidazol-1-one (11C-3g) Area % Report
Data File: PM.dat Method: Acquired: Printed:
C:\EZChrom Elite\Enterprise\Projects\Kaisa\Antonio\ADV-01-007-KH_211-1-2017
3-35-53
C:\EZChrom Elite\Enterprise\Projects\Default\Method\Kaisa\2017_02_14.met 11/1/2017 3:37:21 PM 11/23/2017 12:20:36 PM
4000
0
1
5.017
2000
Volts
2.147
2.010
1.833
Volts
1.460
Retention Time
2
3
4
5
6
7
Minutes
50
0
1
2
2.633
50
0 3
4
5
6
7
Minutes
VWD: Signal A, 254 nm Results Retention Time
Area
Area %
Height
Height %
1.460 1.833 2.010 2.147 5.017
72400266 9279185 9257115 62909053 242833254
18.25 2.34 2.33 15.86 61.22
18100057 2889935 3254113 16507524 64120951
17.26 2.76 3.10 15.74 61.14
S109
mV
100
Retention Time
2.300
mV
100
Totals
Agilent 35900E Interface Radio Results Retention Time 2.300 2.633
Area 35897820 856759
Area % 97.67 2.33
Height 10121351 213881
Height % 97.93 2.07
Totals 36754579
100.00
S110
10335232
100.00
[11C]-1-((5-(2-methylthiazol-4-yl)thiophen-2-yl)methyl)tetrahydropyrimidin-2(1H)-one (11C-3c)
Area % Report
Data File: PM.dat Method: Acquired: Printed:
C:\EZChrom Elite\Enterprise\Projects\Kaisa\Antonio\ADV-01-004-KH10-27-2017
6-42-24
C:\EZChrom Elite\Enterprise\Projects\Default\Method\Kaisa\2017_02_14.met 10/27/2017 6:43:43 PM 11/23/2017 12:14:24 PM 4000
5.017
Volts
2000
3.443
2.210
Volts
3.060
Retention Time
0
1
2
3
4
5
6
7
Minutes
Retention Time
25
25
3.133
0.617
mV
50
mV
50
0 0
1
2
3
4
5
6
7
Minutes
VWD: Signal A, 254 nm Results Retention Time
Area
Area %
Height
Height %
2.210 3.060 3.443 5.017
29926977 95922426 36962816 293892327
6.55 21.00 8.09 64.35
5904414 32599216 9493374 67175799
5.13 28.30 8.24 58.33
S111
Totals
Agilent 35900E Interface Radio Results Retention Time 0.617 3.133
Area 317335 21034778
Area % 1.49 98.51
Height 78249 5821749
Height % 1.33 98.67
Totals 21352113
100.00
S112
5899998
100.00
[11C] 1-(thiophen-2-ylmethyl)tetrahydropyrimidin-2(1H)-2-one (11C-3b) Area % Report Data File: C:\EZChrom Elite\Enterprise\Projects\Kaisa\Antonio\ADV-01-005-KH_210-312017 4-16-43 PM.dat Method: C:\EZChrom Elite\Enterprise\Projects\Default\Method\Kaisa\2017_02_14.met Acquired: 10/31/2017 4:18:07 PM Printed: 11/23/2017 12:18:07 PM 4000
5.017
2.603
2000
Volts
Volts
Retention Time
0
1
2
3
4
5
6
7
Minutes
Retention Time
0
10
2.667
10
0.883
20
1
2
3
4
5
6
7
Minutes
VWD: Signal A, 254 nm Results Retention Time
Area
Area %
Height
Height %
2.603 5.017
23223648 244916125
8.66 91.34
7780423 63985571
10.84 89.16
Totals
S113
mV
30
20
0.633
mV
30
Agilent 35900E Interface Radio Results Retention Time 0.633 0.883 2.667
Area 575322 2146319 12477499
Area % 3.79 14.12 82.09
Height 127168 90522 3029581
Height % 3.92 2.79 93.30
Totals 15199140
100.00
S114
3247271
100.00
[11C] 5-iodo-1H-benzo[d]imidazol-2(3H)-one (11C-2d)
S115
116
[11C]-CGP-12177 (11C-4e)
117
1.862 1.898
1.606
0.800
[11C] 1H-benzo[d]imidazol-2(3H)-one (11C-2a)
118
[11C] 5-chloro-1H-benzo[d]imidazol-2(3H)-one (11C-2b)
119
[11C] 5-fluoro-1H-benzo[d]imidazol-2(3H)-one (11C-2c)
120
0.965
2.653
3.580
2.404
1.356
[11C] 5-methoxy-1H-benzo[d]imidazol-2(3H)-one (11C-2e)
121
3.000
0.867
2.143
1.648
1.440
0.612
2.066
[11C] tert-butyl (2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)carbamate (11C-2f)
122
[11C] methyl 2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-carboxylate (11C-2g)
123
1.029
2.258
0.603
2.225
2.821
[11C] 5-(trifluoromethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2h)
124
[11C] 1H-naphtho[1,2-d]imidazol-2(3H)-one (11C-2i)
125
[11C] 4,6-dimethyl-1H-benzo[d]imidazol-2(3H)-one (11C-2j)
126
0.933
2.625
0.759
2.467
[11C] 4-phenyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (11C-2k)
127
0.923
1.713
3.223
0.736
[11C] 1-benzyl-1H-benzo[d]imidazol-2(3H)-one (11C-2l)
128
0.800
2.737
0.630
4.083
3.295
[11C] 1-(naphthalen-2-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2m)
129
0.967
2.027
0.689
1.992
2.961
[11C] 1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-benzo[d]imidazol-2(3H)-one (11C-2n)
130
[11C] 1-benzyltetrahydropyrimidin-2(1H)-one (11C-3a)
131
[11C] 7-methyl-1-(5-(2-oxotetrahydropyrimidin-1(2H)-yl)hexyl)-3,4,5,7-tetrahydro-1H-purine-2,6dione (11C-3e )
132
[11C] Oxatomide (11C-4a)
133
[11C] Domperidone (11C-4b)
134
[11C] 5HT7-receptor antagonist (11C-4c)
135
2.942
1.891
0.590
1.813
[11C] Flibanserin (11C-4d)
136
137
1.681
9.413
0.930
25.684
1.551 3.061
V. Radio-TLC Analysis for 14C-Labeled Compounds
138
[14C]-NR2-receptor antagonist (14C-4c)
139
[14C] Domperidone (14C-4b)
140
[14C] Flibanserin (14C-4d)
141
[14C] Oxatomide (14C-4a)
142
