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Benzyl bromide

Benzyl bromide is a versatile organic compound used in a variety of chemical reactions and applications.
It is a colorless liquid with a pungent odor, and is commonly used as a precursor in the synthesis of other benzyl compounds.
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Publication 2011
1H NMR 2-(dimethylamino)ethyl methacrylate Anabolism Atmosphere Bromides Chloroform Ethanol MLL protein, human Powder Pulse Rate Radionuclide Imaging Recycling Solvents Spectroscopy, Fourier Transform Infrared Vacuum Viscosity

Material and reagents: Testosterone (17β‐hydroxy‐4‐androsten‐3‐one), yeast extract, ABTS and naphthalene were purchased from AppliChem (Arheilgen, Germany); H2O2 (30 % w/v), soybean peptone, agar, α‐d‐glucose, benzyl alcohol, phenol, and sodium azide were from Carl Roth; malt extract, 2‐chlorophenol, and 4‐chlorophenol were obtained from Merck; glucose oxidase from Aspergillus niger was purchased from Sigma–Aldrich (specific activity 215 U mg−1). All other chemicals were purchased from Sigma–Aldrich at the highest purity available.
Peroxygenases of A. aegerita (AaeUPO) and M. rotula (MroUPO) were produced and purified as described previously;14, 16 recombinant UPOs from C. cinerea (rCciUPO) and H. insolens (rHinUPO=rnovo) were gifts from Novozymes A/S (Copenhagen, Denmark).25, 26, 27, 28The specific activities of AaeUPO and MroUPO were 63.5 U mg−1 and 48.1 U mg−1, respectively (1 U is the oxidation of 1 μmol veratryl alcohol to veratraldehyde per 1 min at 23 °C).14Cultivation ofC. globosum: C. globosum (strain DSM 62110) was purchased from the German Collection of Microorganisms and Cell cultures (Braunschweig, Germany) and was routinely grown on malt extract agar medium (malt extract (20 g L−1) and agar (15 g L−1)) at 24 °C. For enzyme production, the fungus was cultured in 500 mL Erlenmeyer flasks containing carbon‐ and nitrogen‐rich basic liquid medium (200 mL; glucose (42 g L−1), peptone (18 g L−1), yeast extract (4.5 g L−1) in deionized water) on a rotary shaker (120 rpm) at 24 °C for four weeks. Liquid cultures were inoculated with a mycelial suspension (5 % v/v) obtained by homogenization of the content of two agar plates fully covered with fungal mycelium in sterile sodium chloride (100 mL, 0.9 % w/v).
Enzyme assays: UPO activities were measured photometrically by monitoring the oxidation of veratryl alcohol (5 mm) into veratraldehyde at 310 nm (ϵ310=9300 m−1 cm−1) in McIlvaine buffer at pH 7.14 Reaction was started by the addition of hydrogen peroxide (2 mm). Laccase activity during cultivation was determined by the oxidation of ABTS to the corresponding ABTS cation radical at 420 nm (ϵ420=36 000 m−1 cm−1) in McIlvaine buffer at pH 4.5 in the absence of H2O2.29 The specific ring‐hydroxylating activity of CglUPO was monitored by the oxygenation of naphthalene (1 mm) to naphthalene oxide and 1‐naphthol at 303 nm (ϵ303=2030 m−1 cm−1) in McIlvaine buffer at pH 6.0; the reaction was started by adding hydrogen peroxide (2 mm).30Purification and characterization ofCglUPO: All purification steps were carried out at room temperature. Enzyme fractions were assayed for UPO activity, and protein content was determined with a Pierce BCA protein assay kit (Thermo Fisher) with bovine serum albumin as standard. Protein purification was carried out by using ammonium sulfate precipitation and fast protein liquid chromatography (FPLC) on Q‐Sepharose FF (IEC), Superdex75 (SEC), and Mono Q columns (IEC), successively. All chromatographic steps were accomplished with an ÄKTA purifier FPLC system (GE Healthcare).
The molecular mass of purified CglUPO was analyzed by SDS‐PAGE by using a 10 % Bolt Bis‐Tris Gel (Thermo Fisher Scientific). The separated protein bands were visualized with a Colloidal Blue Staining Kit (Generon Ltd, Berkshire, UK, order code GEN‐QC‐Stain‐1L); a protein marker (#26616, Thermo Fisher Scientific) was used as standard.
Proteomic enzyme identification was performed at the Helmholtz‐Centre for Environmental Research—UFZ, Department of Molecular Systems Biology (Leipzig, Germany). For detailed information (peptide mapping), see the Supporting Information.
Kinetic constants (Km, kcat) of CglUPO and pH optima were determined for veratryl alcohol, benzyl alcohol, DMP, ABTS, NBD (pH 7),31 and naphthalene (pH 6; Supporting Information). Halogenating activity was tested by incubating CglUPO (0.2 U mL−1, 0.46 μm) in potassium phosphate buffer (100 mm, pH 3 and pH 7) in the presence of phenol (0.1 mm), potassium bromide or chloride (10 mm) and H2O2 (2 mm).32 After 10 min, the reaction mixture was analyzed by HPLC for the formation of bromo‐ and chlorophenols against authentic standards.
Enzymatic conversion of testosterone: The reaction mixture (total volume 0.5 mL) contained purified AaeUPO (2 U mL−1, 0.7 μm), MroUPO (2 U mL−1, 1.3 μm) or CglUPO (0.2 U mL−1, 0.46 μm) in potassium phosphate buffer (20 mm, pH 7) with testosterone (5 mm), α‐d‐glucose (2 %, w/v), and acetone (5 %, v/v). Reactions were started by addition of glucose oxidase (GOx, 0.02 U mL−1) and stirred at room temperature for 24 h (after this time, no residual activity of CglUPO was detectable). Kinetic data were determination for CglUPO (2 U mL−1, 4.8 μm) with testosterone (5 mm) in potassium phosphate buffer (20 mm, pH 7). Reactions were initiated by the addition of hydrogen peroxide (2 mm) and stopped after 2 min by adding sodium azide (1 mm). Higher concentrations of hydrogen peroxide were not applied, in order to prevent enzyme inactivation from heme bleaching and the disproportionate increase in the UPO intrinsic catalase activity (both have been reported for other UPOs and heme peroxidases).33, 34, 35 Products were recovered with reversed‐phase SPE cartridges (Strata‐X 33u, Phenomenex), with elution in methanol, and analyzed by HPLC.
At preparative scale, a 500 mL flask was filled with testosterone (100 mg, 0.35 mmol), acetone (10 mL), water (140 mL), potassium phosphate buffer (40 mL 0.1 m, pH 7), and CglUPO stock solution (20 mL 400 U in potassium phosphate buffer (0.1 m, pH 7)). The reaction mixture was stirred at room temperature while hydrogen peroxide (100 mm, 4 mL h−1) was continuously supplied by a syringe pump. Hydrogen peroxide was used instead of glucose/GOx, in order to ensure constant peroxide dosage and to avoid impurities in the reaction mixture (glucose and gluconolactone); the syringe pump system was as effective as the GOx‐based H2O2 generation system. Samples (50 μL) were taken from the reaction mixture every 30 min, and the reaction (in the samples) was stopped by adding acetonitrile (50 μL) and sodium azide (10 μL, 10 mm). The samples were centrifuged, and the supernatants were analyzed by HPLC (below). After 7 h, thin layer chromatography (in ethyl acetate/n‐hexane, 9:1) indicated complete conversion of testosterone. The reaction mixture was extracted three times with ethyl acetate (50 mL), then the combined organic fractions were dried with Na2SO4 and evaporated to dryness to give 91 mg of crude products 1 a (Rf 0.67) and 1 b (Rf 0.11). The compounds were purified by chromatography on silica gel with ethyl acetate/n‐hexane (9:1) as the eluent to obtain 65 mg (61.1 %) of 1 a (96.3 % purity) and 7 mg (6.6 %) of 1 b (98.7 % purity).
Analytical methods: The HPLC‐MS system (Waters) comprised a 2690 separation module, a 2996 photo diode array detector, and a Micromass ZMD 2000 single quadrupole mass spectrometer. Separation was on a LiChrospher C18 column (125×4 mm, 5 μm, Phenomenex) with mobile phases A (formic acid (0.1 %)) and B (acetonitrile) and at stepwise gradient (20 % B (3 min), increase to 55 % B (20 min), increase to 90 % B (3 min)). The final level was maintained until all analytes had been eluted from the column (flow‐rate 1 mL min−1, column temperature 30 °C). Reaction products were identified by comparison to authentic standards based on retention time, UV absorption spectrum and mass spectra [M+H]+ or [M−H] ions, and quantified by total peak area by using response factors of the same or similar compounds. Data from replicates were averaged. Standard deviations were below 5 % of the mean in all cases.
1H (400 MHz) and 13C (100 MHz) NMR spectra of testosterone and its enzymatic conversion products were obtained on Bruker spectrometer (Bruker Avance II 400 MHz) in the solvent indicated.
Publication 2017

Chemistry All chemical compounds were purchased from commercial suppliers of Merck and Aldrich companies. The purity of the prepared compounds was proved by thin layer chromatography (TLC) using various solvents of different polarities. Merck silica gel 60 F254 plates were applied for analytical TLC. Column chromatography was performed on Merck silica gel (70-230 mesh) for purification obtained compounds. 1H-NMR spectra were recorded using a Brucker 200 MHz spectrometer, and chemical shifts were expressed as δ (ppm) with tetramethylsilane (TMS) as internal standard. The IR spectra were obtained on a Shimadzu 470 spectrophotometer (potassium bromide disks). Melting points were determined using electrothermal melting point analyzer apparatus and were uncorrected. The mass spectra were run on a Finigan TSQ-70 spectrometer (Finigan, USA) at 70 eV. All cell lines were purchased from the Pasteur Institute of Iran.
According to the Figure 3, 5-amino- 1,3,4-thiadiazole-2-thiol (1 (link)) was treated with 4-trifluoromethylphenylacetic acid for direct coupling of acid with amine. The reaction was carried out in the presence of EDC and hydroxybenzotriazole (HOBt) in acetonitrile as solvent. The termination of reaction was proved by thin layer chromatography (TLC). After completion, the solvent was evaporated using rotary evaporator apparatus and ethyl acetate and water were added. The aqueous phase was removed and the organic phase was washed two times by sodium bicarbonate 5%, diluted sulfuric acid and brine (16 (link)-19 ). Anhydrous sodium sulfate was added for drying and filtration was done. Ethyl acetate was removed under reduced pressure and a yellow powder was obtained. The obtained product was used after crystallization from ethanol for the next step. Various benzyl chloride derivatives were reacted with compound 2 for obtaining the final appropriate products (3a-3l). The 1H NMR, IR and MS spectra were used to confirm the synthesized compounds.
Synthesis of N-(5-Mercapto-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (2)In a flask, equimolar amounts of 4-trifluoromethylphenylacetic acid, EDC and HOBtin acetonitrile solvent were stirred for 30 min and then equimolar quantity of 5-amino-1,3,4-thiadiazole-2-thiol was added. The stirring condition was continued for 24 h. The end point of the reaction was determined by thin layer chromatography(TLC). Acetonitrile was removed under reduced pressure and ethyl acetate/water was added. The aqueous layer was removed and organic layer was washed two times by sodium bicarbonate 5%, diluted sulfuric acid and brine. Anhydrous sodium sulfate was added for drying and then filtered. The ethyl acetate was evaporated using rotary evaporator apparatus. The obtained yellowish solid was washed by dry ether and used for the next step.
mp. 171°C, Yield: 65%,C11H8F3N3OS2, MW: 319 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 3.76 (s, 2H, -CH2CO-), 3.95 (s, 1H, -SH), 7.58 (m, 2H, J = 8Hz), 8.19 (m, 2H, J = 8Hz, 4-trifluoromethylphenyl), 12.76 (brs, 1H, NH).IR (KBr, cm-1) ῡ: 3265, 1697, 1580, 1519, 1321, 1155, 1103, 1064, 821, 705.MS(m/z, %): M++2: 322(10), M+: 320(10), 279(45), 276(35), 167(80), 159(95), 149(100), 133(15), 109(15), 71(15), 57(20).
General procedure for synthesis of compounds 3a-3lEquimolar quantities of appropriate benzyl chloride derivative was treated with 2-(4-fluorophenyl)-N-(5-mercapto-1,3,4-thiadiazol-2-yl)acetamide (2 (link)). Equimolar amount of potassium hydroxide in absolute ethanol was added to convert the thiol moiety to the thiolate anion. Then, the related benzyl chloride derivative was added to the reaction medium and reflux condition was performed for 24 h. Crushed ice was added and the precipitate filtered, washed by cool water and purified by appropriate procedures such as crystallization or column chromatography (EtOAC/Petroleum ether: 3/2).
N-(5-(2-Fluorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3a)mp. 201 °C, Yield: 42%,C18H13F4N3OS2, MW: 427 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 4.00 (s, 2H, -CH2CO-), 4.53 (s, 2H, -CH2S-), 7.16-7.54 (m, 2-fluorobenzyl), 7.59 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.76 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 13.00 (s, NH). IR(KBr, cm-1) ῡ: 3158, 3050, 2898, 1690, 1560, 1493, 1456, 1421, 1402, 1337, 1301, 1236, 1168, 1117, 1071, 1019, 972, 844, 757, 693, 655.MS(m/z, %): M+: 427(95), 377(60), 325(35), 297(30), 236(25), 236(15), 159(75), 109(100).
N-(5-(3-Fluorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3b)mp. 190 °C, Yield: 58%,C18H13F4N3OS2, MW: 427 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 3.93 (s, 2H, -CH2CO-), 4.53 (s, 2H, -CH2S-), 7.01-7.47 (m, 3-fluorobenzyl), 7.58 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.98 (s, NH).IR(KBr, cm-1) ῡ: 3448, 3156, 2916, 1699, 1562, 1488, 1358, 1326, 1172, 1110, 1068, 837.MS(m/z, %): M+: 427(85), 377(80), 325(40), 297(25), 236(20), 159(70), 109(100).
N-(5-(4-Fluorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3c)mp. 166 °C, Yield: 40%,C18H13F4N3OS2, MW: 427 g/mol,1H NMR (DMSO-d6 , 200 MHz) δ: 3.99 (s, 2H, -CH2CO-), 4.51 (s, 2H, -S-CH2-), 7.24 (t, 2H, 4-fluorobenzyl), 7.48 (t, 2H, 4-fluorobenzyl), 7.58 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.97 (s, NH).IR(KBr, cm-1) ῡ: 3370, 3030, 2920, 2829, 1701, 1558, 1508, 1323, 1219, 1160, 1109, 1060, 835.MS(m/z, %): M+: 427(90), 377(75), 325(40), 297(30), 236(25), 159(75), 109(100).
N-(5-(2-Chlorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3d)mp. 208 °C, Yield: 54%,C18H13ClF3N3OS2, MW: 443 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 4.00 (s, 2H, -CH2CO-), 4.59 (s, 2H, -CH2S-), 7.35 (m, 2-chlorophenyl), 7.48 (d, 2H, J = 12 Hz, 4-trifluoromethylphenyl), 7.55 (m, 2-chlorophenyl), 7.97(d, 2H, J = 12 Hz, 4-trifluoromethylphenyl), 13.00 (s, NH). IR(KBr, cm-1) ῡ: 3447, 3165, 2922, 1700, 1570, 1445, 1326, 1171, 1109, 1067, 868, 759.MS(m/z, %): M+:443(10), 410(55), 409(85), 159(40), 127(50), 125(100), 109(10), 89(10).
N-(5-(3-Chlorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3e)mp. 198 °C, Yield: 57%,C18H13ClF3N3OS2, MW: 443 g/mol,1H NMR (DMSO-d6 , 200 MHz) δ: 4.00 (s, 2H, -CH2CO-), 4.59 (s, 2H, -S-CH2-), 7.33-7.40 (m, 3-chlorobenzyl), 7.43 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.56 (m, 3-chlorobenzyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 13.00 (s, NH).IR(KBr, cm-1) ῡ: 3427, 3165, 2914, 2730, 1700, 1569, 1446, 1356, 1326, 1301, 1170, 1112, 1066, 868, 839, 759.MS(m/z, %): M+:443(15), 410(60), 409(65), 282(10), 159(55), 127(35), 125(100), 89(10).
N-(5-(4-Chlorobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3f)mp. 198 °C, Yield: 34%,C18H13ClF3N3OS2, MW: 443 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 3.99 (s, 2H, -CH2CO-), 4.52 (s, 2H, -S-CH2-), 7.38-7.48 (m, 4-chlorobenzyl), 7.58 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.78 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.97 (s, NH).IR(KBr, cm-1) ῡ: 3440, 3130, 3040, 2877, 1690, 1556, 1334, 1163, 1118, 1068, 1020, 846, 745, 702.MS(m/z, %): M++1: 444(10), M+:443(10), 410(60), 409(80), 282(12), 178(12), 159(40), 127(50), 125(100), 109(10), 89(10).
N-(5-(3-Methoxybenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3g)mp. 160 °C, Yield: 32%,C19H16F3N3O2S2, MW: 439 g/mol,1H NMR (DMSO-d6 , 200 MHz) δ:1H NMR (DMSO-d6 , 200 MHz) δ: 3.80 (s, 3H, -OCH3), 3.99 (s, 2H, -CH2CO-), 4.48 (s, 2H, -S-CH2-),7.06(m, 3H, 3-methoxybenzyl), 7.32(m, 1H, 3-methoxybenzyl), 7.58(d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.97 (s, NH).IR(KBr, cm-1) ῡ:3167, 3038,2946, 1735, 1702, 1607, 1577, 1488, 1438, 1358, 1325, 1302, 1271, 1158, 1113, 1068, 839, 777, 736.MS(m/z, %): M+: 439(40), 159(75), 122(45), 121(100), 109(35).
N-(5-(4-Methoxybenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3h)mp. 219 °C, Yield: 36%,C19H16F3N3O2S2, MW: 439 g/mol,1H NMR (DMSO-d6 , 200 MHz) δ: 3.76 (s, 3H, -OCH3), 3.99 (s, 2H, -CH2CO-), 4.46 (s, 2H, -S-CH2-), 6.92 (d, 2H, J = 8 Hz, 4-methoxybenzyl), 7.35 (d, 2H, J = 8 Hz, 4-methoxybenzyl), 7.58 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.96 (s, NH).IR(KBr, cm-1) ῡ: 3265, 3045, 2870, 1691, 1554, 1510, 1400, 1332, 1298, 1170, 1122, 1107, 1066, 827. MS(m/z, %): M++2: 441(15), M++1: 440(20), M+: 439(25), 159(60), 122(45), 121(100), 109(30).
N-(5-(2-Nitrobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3i)mp. 146 °C, Yield: 35%,C18H13F3N4O3S2, MW: 454 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 4.00 (s, 2H, -CH2CO-), 4.79 (s, 2H, -CH2S-), 7.49 (m, aromatic), 8.11 (m, aromatic), 8.21 (m, aromatic), 13.00 (s, NH). IR(KBr, cm-1) ῡ: 3447, 3165, 2923, 1698, 1612, 1573, 1527, 1443, 1335, 1168, 1106, 1066, 830, 827, 702.MS(m/z,%): M+: 454(15), 270(35), 242(65), 225(15), 195(100), 179(65), 165(85), 136(70), 106(45), 90(50), 78(35).
N-(5-(3-Nitrobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3j)mp. 132 °C, Yield: 45%,C18H13F3N4O3S2, MW: 454 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 3.84 (s, 2H, -CH2CO-), 4.51 (s, 2H, -CH2S-), 7.71(m, 5H, aromatic), 8.16(m, 3H, aromatic), 13(brs, NH).IR(KBr, cm-1) ῡ: 3318, 3154, 2850, 1692, 1629, 1562, 1527, 1508, 1487, 1348, 1329, 1163, 1117, 1073, 810, 747.MS(m/z,%): M++1: 455(7), M+: 454(15), 270(30), 242(75), 195(100), 179(70), 165(90), 136(60), 106(35), 90(45), 78(60).
N-(5-(4-Nitrobenzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3k)mp. 198 °C, Yield: 54%,C18H13F3N4O3S2, MW: 454 g/mol,1H NMR (DMSO-d6, 200 MHz) δ: 3.99 (s, 2H, -CH2CO-), 4.66 (s, 2H, -S-CH2-),7.55(m, 2H, aromatic), 7.77(m, 4H, aromatic), 8.25(m, 2H, aromatic), 12.99(brs, NH).IR(KBr, cm-1) ῡ:3265, 3153, 3040, 2912, 2852, 1697, 1554, 1517, 1342, 1323, 1155, 1103, 1064, 1020, 960, 830.MS(m/z,%): M++1: 455(10), M+: 454(10), 270(40), 242(75), 225(40), 195(100), 179(95), 165(85), 136(60), 106(50), 90(50), 78(60).
N-(5-(Benzylthio)-1,3,4-thiadiazol-2-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (3l)mp. 203 °C, Yield: 47%,C18H14F3N3OS2, MW: 409 g/mol,1H NMR (DMSO-d6 , 200 MHz) δ: 3.99 (s, 2H, -CH2CO-), 4.52 (s, 2H, -S-CH2-), 7.30-7.49 (m, 5H, benzyl), 7.58 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 7.75 (d, 2H, J = 8 Hz, 4-trifluoromethylphenyl), 12.97 (s, NH).IR(KBr, cm-1) ῡ: 3375, 3040, 1701, 1556, 1508, 1323, 1294, 1220, 1159, 1107, 1066, 1020, 827, 700.MS(m/z, %): M+: 410(100), 409(95), 408(95), 159(75), 148(60), 91(75).
MTT assayDiverse derivatives of 1,3,4-thiadiazole (compounds 3a-3l) were tested for cytotoxic activity at 0.1-250 μg/mL concentration in three human cancer cell lines of PC3 cell (prostate cancer), U87 (gliobalstoma) and MDA (breast cancer). Cells from different cell lines were seeded in 96-well plates at the density of 8000–10,000 viable cells per well and incubated for 48 h to allow cell attachment.The cells were then incubated for another 48-96 h (depends to cell cycle of each cell line) with various concentrations of compounds 3a-3l. Cells were then washed inPBS, and 20 μL of MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide solution (5 mg/mL) were added to each well. An additional 4 h of incubation at 37°C were done, and then the medium was discarded. Dimethyl sulfoxide (60 μL) was added to each well, and the solution was vigorously mixed to dissolve the purple tetrazolium crystals. The absorbance of each well was measured by plate reader (Anthous 2020; Austria) at a test wavelength of 550 nm against a standard reference solution at 690 nm. The amount of produced purple formazan is proportional to the number of viable cells (16 (link)).
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Acetic Acid Anions benzyl bromide Disulfides Ethers Mesylates phthalimide poly(ethylene glycol)-block-poly(propylene sulfide) Polymerization Polymers propylene sulfide Sulfhydryl Compounds thioacetic acid
All reagents and solvents
were obtained from commercial sources and used without further purification.
All reactions, unless otherwise stated, were performed under a nitrogen
atmosphere. Reactions were monitored using either aluminum or glass-backed
silica TLC plates impregnated with a fluorescent indicator, absorbing
at 254 nm. Silica gel column chromatography was performed on a CombiFlash
Rf Teledyne ISCO system using hexane, ethyl acetate, methylene chloride,
or methanol as eluent. Reverse phase column chromatography (C18 column)
was performed on the same instrument using 0.1% formic acid in methanol,
acetonitrile, or water as eluent. Separations were monitored by mass
spectrometry via a Teledyne ISCO RF+ PurIon ESI-MS or APCI-MS
detector with 1 Da resolution. The purity of all compounds used in
assays was determined to be ≥95% by 1H NMR spectroscopy
and confirmed by high-resolution mass spectrometry (HRMS) experiments
using an Agilent 6230 Accurate-Mass LC-TOFMS located in the University
of California, San Diego, Molecular Mass Spectrometry Facility (MMSF).
Standard resolution MS was performed at either the aforementioned
MMSF or Teledyne ISCO RF+ PurIon MS. Microwave reactions
were performed using a CEM Discover series S-class microwave reactor
in pressure-sealed vessels. Docking simulations were performed using
MOE, version 2014.0901. Flexible receptor modeling (induced fit) was
employed in these simulations, and metal-binding atoms were fixed.
Synthesis of 5-hydroxy-2-methyl-4-oxo-4H-pyran-3-carboxamide
derivatives is outlined in Scheme 1. Bromopyruvic acid was transformed to bromodiethoxypropanoic
acid by treatment with triethyl orthoformate in the presence of catalytic
sulfuric acid. The acetal-protected propanoic acid was then activated
as the p-nitrophenyl ester 74. Pyrone
ring formation was achieved over two steps by the slow addition of 74 to a solution of ethyl or tert-butyl acetoacetate
that was previously deprotonated by sodium hydride. After the initial
nucleophilic attack of the activated ester by the acetoacetate, ring
closing was accomplished via nucleophilic addition by heating the
reaction mixture to reflux for 4–6 h. The pyrone-acetal 75a was deesterified by stirring briefly with trifluoroacetic
acid (TFA) in CH2Cl2. The tert-butyl ester was removed selectively by controlling the time of the
reaction, as the ester is more labile in the absence of water than
the acetal. Key intermediate 76 was used to prepare various
amides using analogous conditions; specifically, 76 was
activated with HATU and triethylamine in DMF. Addition of the amine
was followed by heating and stirring for ∼18 h at 60 °C.
After isolation of the formed amide the acetal was deprotected in
water and acid to reveal the 3-keto intermediate, which rapidly tautomerizes
to form the desired 3-hydroxide species.
To acquire SAR at the 6-position, a library of amine derivatives
was prepared from commercially available kojic acid (Scheme 2). The phenolic oxygen of 8 was selectively protected as a benzyl ether by treatment
with benzyl bromide in the presence of potassium carbonate in DMF
at 80 °C. Compound 78 was prepared quantitatively
by reacting 77 with thionyl chloride. Nucleophilic addition
of various primary and secondary amines followed by selective hydrolysis
of the benzyl ether in a mixture of TFA, concentrated HCl, and glacial
acetic acid afforded aminomethylpyrones in good yields. Amide
and sulfonamide derivatives were generated by the nucleophilic addition
of sodium azide to 78, followed by reduction of 79 with triphenylphosphine to afford 80 as a
key intermediate. Compound 80 was reacted with various
acid chlorides and sulfonyl chlorides to generate 2831, after deprotection of the phenol by boron trichloride.
Preparation
of pyridinone derivatives of allomaltol (2) possessing N-aryl and -alkyl substituents is described
generally in Scheme 3. Kojic chloride (81) was derived from 8 and reduced to 2 using metallic zinc and strong acid.
While dehydration of the hydroxypyrone ring to afford hydroxypyridinone
derivatives was shown to be possible, benzyl protection of the hydroxyl
group prior to dehydration greatly improved yields and suppressed
formation of side products. Key intermediate 82 was irradiated
in a microwave reactor in the presence of excess amine and acetic
acid to produce a wide variety of pyridinone derivatives. Microwave
heating under increased pressure greatly accelerated reaction rates;
dehydration using conventional heating could be accomplished but required
refluxing reactants in ethanol for a period of days. Removal of the
benzyl ether after dehydration was shown to be accomplished quickly
and efficiently employing boron trichloride as a dealkylating agent
or less efficiently using a 5:5:1 mixture of concentrated hydrochloric
acid, glacial acetic acid, and TFA, as a milder alternative to boron
trihalides (Supporting Information).
Fragment merging
was accomplished as shown in Scheme 4. Merging of N-arylpyridinones with
5-position carboxylates began with advanced
intermediate 76a. Hydrolysis of the acetal followed by
tautomerization yielded 83. After benzyl protection of
the phenol, dehydration was accomplished with microwave heating using
dry ethanol as a solvent. Hydrolysis of only the benzyl ether using
strong acid afforded compounds 6769 in moderate yields; hydrolysis of both ester and ether was not observed.
Further base catalyzed hydrolysis of the ethyl ester proved difficult
and ultimately resulted in the decomposition of the ring system. As
an alternative route, compound 86 was hydrolyzed to the
free carboxylic acid by stirring in 4% KOH and methanol for several
hours. This reaction proceeded almost quantitatively with no apparent
decomposition of the starting material. Compound 86 was
found to be stable to strongly acidic conditions and was hydrolyzed
to 70 using a mixture of strong acids with good yields.
Synthesis of compound 71 began with chloride compound 78. Nucleophilic substitution with 4-chloro-N-methylaniline afforded pyrone 87. Dehydration, as previously
described, yielded 88 in moderate to low yields. Hydrolysis
in the presence of strong acid afforded 71.
Publication 2016

Most recents protocols related to «Benzyl bromide»

Hombikat UV100 (UV100, Venator, Wynyard, UK), sodium hydroxide (NaOH, Fisher Scientific, Hampton, NH, USA), hydrochloric acid (HCl, 37%, Fisher Scientific) and deionized water (DI) were used to synthesize pristine hydrogen titanate. Bismuth bromide (BiBr3, Sigma-Aldrich, St. Louis, MO, USA), cesium bromide (CsBr, Sigma-Aldrich), dimethyl sulfoxide (DMSO, Sigma-Aldrich) and isopropanol (IPA, Sigma-Aldrich) was used to synthesize pristine Cs3Bi2Br9 and Cs3Bi2Br9/Hydrogen Titanate heterostructures. benzyl alcohol (BnOH, Sigma-Aldrich), Acetonitrile (MeCN, Sigma-Aldrich) and 1,2-dichlorobenzene (DCB, Sigma-Aldrich) were used for benzyl alcohol oxidation experiments. Potassium iodide (KI, Sigma-Aldrich), silver nitrate (AgNO3, Sigma-Aldrich), p-benzoquinone (p-BQ, Sigma-Aldrich) and tert-butyl alcohol (TBA, Sigma-Aldrich) were used for scavenger tests to identify the main reactive oxygen species (ROSs) responsible for BnOH conversion. All chemical reagents were used in purities >99% as received without any further purification.
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Publication 2024
PP (thickness of 1 mm) was purchased from
AS ONE (Osaka, Japan). Benzophenone, dimethylformamide, benzyl bromide,
and 4-vinylpyridine were purchased from Tokyo Kasei Kogyo (Tokyo,
Japan). All other reagents were purchased from Fujifilm Wako Pure
Chemical (Osaka, Japan).
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Publication 2024
A piece of PP (10 mm ×
10 mm) precleaned
with acetone. The thus-precleaned sample was drop-coated with a solution
of benzophenone (0.2 g, 1.1 mmol) in 4-vinylpyridine (1.0 g, 9.5 mmol),
covered with a borosilicate cover glass and irradiated with UV light
at λ = 365 nm using a light-emitting-diode device (CCS, Kyoto,
Japan), and placed in a stainless-steel vat. The vat was covered with
a glass plate and purged with nitrogen for 5 min. The specimen was
then irradiated with intensities of 30–170 mW cm–2 for 1–20 min. After irradiation, the sample was sequentially
washed with methanol, acetone, chloroform, and ethyl acetate and then
sonicated in methanol for 10 min. Further, the sample was exposed
to a solution of benzyl bromide (4.4 g, 25.7 mmol) in dimethylformamide
(100 mL) and heated at 90 °C for 0.5–5 h under nitrogen
upon slow stirring to form QACs on the grafted polymer chains. Finally,
the sample was sequentially washed with methanol and acetone, sonicated
in methanol for 10 min, vacuum-dried at 60 °C for 1 h, dipped
in sterilized ultrapure water for 30 min, and air-dried at approximately
25 °C for 1 h on a clean bench. All ultrasonic cleaning was performed
with a bath-type sonicator (Branson M2800-J, Emerson Electric, Missouri)
operating at 110 W and an emission frequency of 40 kHz. Specimens
prepared using a constant irradiation intensity of 170 mW cm–2 (measured under the glass plate covering the vat) and irradiation
durations of 1, 3, and 10 min were denoted as T-1, T-3, and T-10,
respectively. Specimens prepared at a constant irradiation time of
10 min and irradiation intensities of 30, 50, and 100 mW cm–2 were denoted as I-30, I-50, and I-100, respectively. All T- and
I-series specimens were prepared by using a benzylation time of 3
h.
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Publication 2024
Cobalt(II) tetraphenylporphyrin (CoIITPP) (TCI America, >80.0%) was recrystallized from methylene chloride (Fisher Chemical, HPLC) before use: ~1 g of as-received CoIITPP was dissolved in ~500 ml of methylene chloride via sonication for ~2 h; the solution was filtered with 0.45 μm syringe filters (VWR), transferred in a crystallization dish (Pyrex), and evaporated under ambient atmosphere overnight; the recrystallized solid was dried at 60 °C and stored in an Argon-filled glovebox. Tetrabutylammonium hexafluorophosphate (NBu4PF6) (TCI America, >98.0%) was dried at 90 °C under vacuum and stored in the glovebox. Dimethylformamide (DMF) (Sigma-Aldrich, anhydrous, 99.8%) was stored in the glovebox and used as received.
A diverse scope of organic electrophiles was studied in this work. Purchased from Sigma-Aldrich were 1-bromobutane (n-BuBr) (99%), 1-iodobutane (n-BuI) (99%, stabilized with copper), 1-bromo-2-methylpropane (i-BuBr) (99%), dichloromethane (CH2Cl2) (anhydrous, ≥99.8%, stabilized with amylene), chloroacetonitrile (ClCH2CN) (99%), acetonitrile (CH3CN) (anhydrous, 99.8%), benzyl bromide-α,α-d2 (PhCD2Br) (98 atom% D), 4-methylbenzyl bromide (p-Me-PhCH2Br) (97%), 4-(trifluoromethoxy)benzyl bromide (p-F3CO-PhCH2Br) (97%), methyl 4-(bromomethyl)benzoate (p-MeO2C-PhCH2Br) (98%), and 4-(bromomethyl)benzophenone (p-PhOC-PhCH2Br) (96%). Purchased from TCI America were 1-bromohexane (n-HexBr) (>98.0%), 1-bromooctane (n-OctBr) (>98.0%), 2-bromobutane (2-BuBr) (>98.0%), 1-chlorobutane (n-BuCl) (>99.0%), 3-chloropropionitrile (Cl(CH2)2CN) (>98.0%), 4-chlorobutyronitrile (Cl(CH2)3CN) (>97.0%), 5-chlorovaleronitrile (Cl(CH2)4CN) (>97.0%), 4-bromobutyronitrile (Br(CH2)3CN) (>97.0%), and benzyl bromide (PhCH2Br) (>98.0%, stabilized with propylene oxide). Purchased from Oakwood Chemical were neopentyl iodide (Me3CCH2I) (98%), 4-iodobutyronitrile (I(CH2)3CN) (97%), and 1-(bromomethyl)-4-methoxybenzene (p-MeO-PhCH2Br) (stabilized with K2CO3). Purchased from Synquest Laboratories were 4-fluorobenzyl bromide (p-F-PhCH2Br) (98%), 4-(trifluoromethyl)benzyl bromide (p-F3C-PhCH2Br) (98%), and 4-cyanobenzyl bromide (p-NC-PhCH2Br) (98%). Purchased from Cambridge Isotope Laboratories were n-BuBr-d9 (98%) and n-BuI-d9 (98%, stabilized with copper). Purchased from AK Scientific was 1-(bromomethyl)-4-phenoxybenzene (p-PhO-PhCH2Br) (95%). Purchased from Thermo Scientific Chemicals was (1-bromoethyl)benzene (PhCH(CH3)Br) (97%). Among all the organic electrophiles mentioned above, solid chemicals were stored in the glovebox and used as received, whereas liquid chemicals were transferred in a Schlenk flask, evacuated under vacuum on a Schlenk line, brought into the glovebox, and dried over 3 Å molecular sieves (Sigma-Aldrich) before use.
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Publication 2024
A mixture of FA and benzyl bromide in ACN was stirred under reflux for almost 12 h, after the completion of reaction (monitored by TLC), the FA derivative (14) was purified by the help of column chromatography.
Publication 2024

Top products related to «Benzyl bromide»

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Benzyl bromide is a colorless, volatile liquid chemical compound commonly used as a laboratory reagent. It has the chemical formula C6H5CH2Br. Benzyl bromide is primarily utilized in various organic synthesis reactions as an alkylating agent.
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Benzyl alcohol is a colorless organic liquid that is commonly used as a solvent and preservative in various laboratory applications. It has a mild, aromatic odor and is miscible with water, alcohol, and many organic solvents. Benzyl alcohol is a versatile chemical that serves as a key component in numerous laboratory procedures and experiments.
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Ethanol is a clear, colorless liquid chemical compound commonly used in laboratory settings. It is a key component in various scientific applications, serving as a solvent, disinfectant, and fuel source. Ethanol has a molecular formula of C2H6O and a range of industrial and research uses.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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N,N-dimethylformamide is a clear, colorless liquid organic compound with the chemical formula (CH3)2NC(O)H. It is a common laboratory solvent used in various chemical reactions and processes.
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Oleic acid is a long-chain monounsaturated fatty acid commonly used in various laboratory applications. It is a colorless to light-yellow liquid with a characteristic odor. Oleic acid is widely utilized as a component in various laboratory reagents and formulations, often serving as a surfactant or emulsifier.
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Triethylamine is a clear, colorless liquid used as a laboratory reagent. It is a tertiary amine with the chemical formula (CH3CH2)3N. Triethylamine serves as a base and is commonly employed in organic synthesis reactions.
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Chloroform is a colorless, volatile liquid with a characteristic sweet odor. It is a commonly used solvent in a variety of laboratory applications, including extraction, purification, and sample preparation processes. Chloroform has a high density and is immiscible with water, making it a useful solvent for a range of organic compounds.
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Toluene is a colorless, flammable liquid with a distinctive aromatic odor. It is a common organic solvent used in various industrial and laboratory applications. Toluene has a chemical formula of C6H5CH3 and is derived from the distillation of petroleum.

More about "Benzyl bromide"

Benzyl bromide, a versatile organic compound, is widely used in various chemical reactions and applications.
This colorless liquid with a pungent odor is commonly employed as a precursor in the synthesis of other benzyl-based compounds.
Closely related to benzyl bromide are several other important chemicals, such as DMSO (dimethyl sulfoxide), which is a popular solvent, and benzyl alcohol, a common preservative.
Ethanol, FBS (fetal bovine serum), and N,N-dimethylformamide are also frequently encountered in chemical and biological research involving benzyl bromide.
Other relevant compounds include oleic acid, a fatty acid used in emulsions, triethylamine, a common base, as well as chloroform and toluene, which are solvents often used in conjunction with benzyl bromide.
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