The resultant tetrazolium salt was dissolved in 100 L dimethylsulfoxide and read at 570 nm using a microplate reader (Varioskan Flash; Thermo Fisher Scientific, Waltham, MA, USA)

The resultant tetrazolium salt was dissolved in 100 L dimethylsulfoxide and read at 570 nm using a microplate reader (Varioskan Flash; Thermo Fisher Scientific, Waltham, MA, USA). Compounds were tested in triplicate in at least three independent assays. important two pharmacophores of the tubulin polymerization inhibitors. The 4′-nitro benzyl moiety interacted with the region P3 as expected. The 6,7-di-methoxy group formed two hydrogen bonds with Ser178 of tubulin. It is also interesting to see an oxygen atom of the 4′-nitro group formed a hydrogen bond with the amino group at the flexible sidechain of Lys352, which exactly explain why the introduction of 4′-nitro benzyl to the C-4 position of the isoquinoline ring of the lead compounds is favorable to activity. Open in a separate window Physique 4 The hypothetical binding mode of the isomer of compound 32 to tubulin protein. P1 and P2 are the two hydrophobic pockets, and P3 is usually a polar region in the interface between /-tubulin. The physique was generated using PyMol (http://pymol.souceforge.net/). 3. Experimental Section 3.1. Chemistry The melting point was determined on a XT4A microscope melting-point apparatus (Keyi Electron Optical Instrument Factory, Beijing, China) without correction.1H NMR and 13C NMR spectra were recorded on BRUKER AVANCE 300 and 600 spectrometers (Bruker Company, Rheinstetten, Germany), with TMS as an internal standard and CDCl3 as the solvent. ESI mass spectra were performed on an API-3000 LC-MS spectrometer (Applied Biosystems, Toronto, ON, Canada). Flash column chromatography was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical, Qingdao, China). All solvents and reagents were purchased from commercial suppliers and, when necessary, were purified and dried by standard protocols. Organic solutions were dried over anhydrous sodium sulfate. The purity of the final compounds was assessed with an Agilent 1200 HPLC (Agilent Technologies, Santa Clara, CA, USA), and the results were greater than 95%. (= 1.5, 4.8 Hz), 8.75 (dd, 2H, = 1.5, 4.5 Hz). The synthetic methods for the intermediates 4bCd were similar to the synthesis of intermadiate 4a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propanenitrile (5a) A mixture of 4a (0.62 g, 2.1 mmol), NaBH4 (0.32 g, 8.5 mmol) and 20 mL MeOH was heated under 50 C for 0.5 h. The mixture was evaporated and the residue diluted with 25 mL EtOAc. The organic layer was dried and filtered and the solvent removed by evaporation. After the solution was cooled and stayed overnight to give the compound 5a (2.18 g, 86.21%) as a white crystals. mp 129C130 C. 1H NMR (300 MHz, CDCl3): 3.10C3.24 (m, 2H), 3.82 (s, 6H), 3.85 (s, 3H), 3.99 (t, 1H), 6.40 (s, 2H), 7.09 (d, 2H, = 4.5 Hz), 8.56 (d, 2H, = 4.5 Hz). The synthetic methods for the intermediates 5bCd were similar to the synthesis of intermadiate 5a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propan-1-amine (6a) BF3O(C2H5)2 (7.5 mmol) was slowly added to a stirred solution of 5a (0.76 g, 2.5 mmol) and NaBH4 (10 mmol) in THF (10 mL) at 0 C. The solution was refluxed for 1 h, poured into water, and extracted with EtOAc (15 mL 3). The combined extracts were dried over anhydrous Na2SO4 and filtered. The solvents were removed by evaporation to afford 6a (0.72 g, 96.61%) as a yellow oil. 1H NMR (300 MHz, CDCl3): 2.96C2.79 (m, 5H), 3.81 (s, 6H), 3.83 (s, 3H), 6.32 (s, 2H), 6.98 (dd, 2H, = 1.5, 4.5 Hz), 8.43 (dd, 2H, = 1.5, 4.5 Hz). The synthetic methods for the intermediates 6bCd were similar to the synthesis of intermadiate 6a. 3,4-Dimethoxy-= 1.5, 4.2 Hz). The synthetic methods for the intermediates 8C19 were similar to the synthesis of intermadiate 7. 1-(3,4-Dimethoxyphenyl)-6,7,8-trimethoxy-4-(pyridin-4-ylmethyl)-3,4-dihydroisoquinoline (20) A mixture of 7 (0.70 g, 1.5 mmol), POCl3 (0.82 mL, 9 mmol) and CH3CN (15 mL) was stirred and heated under reflux for 4 h, then the solvents CD48 were removed by evaporation, and the residue was dissolved in EtOAc (30 mL). Then the solution was neutralied to pH = 7 with saturated aqueous Na2CO3 and washed by water (30 mL 3). The organic layer was dried over anhydrous MgSO4 and filtered. Then the filtrate was concentrated under reduced pressure, The residue after evaporation was purified by flash chromatography on silica gel (eluent: CH2Cl2/MeOH = 100:1 = 4.8, 14.7 Hz), 3.76 (s, 3H), 3.80 (s, 3H), 3.93 (s, 3H), 3.94 (s, 3H), 4.04 (dd, 1H, = 3.0, 14.4 Hz), 6.20 (s, 1H), 6.88 (d, 1H, = 8.4 Hz), 7.02 (dd, 1H, = 1.8, 5.1 Hz), 7.07 (dd, 2H, = 1.5, 4.5 Hz), 7.17 (d, 1H, = 1.8 Hz), 8.51 (dd, 2H, = 1.5, 4.5 Hz). ESI-MS (=.Flash column chromatography was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical, Qingdao, China). group formed two hydrogen bonds with Ser178 of tubulin. It is also interesting to see an oxygen atom of the 4′-nitro group formed a hydrogen bond with the amino group at the flexible sidechain of Lys352, which exactly explain why the introduction of 4′-nitro benzyl to the C-4 position of the isoquinoline ring of the lead compounds is favorable to activity. Open in a separate window Physique 4 The hypothetical binding mode of the isomer of compound 32 to tubulin protein. P1 and P2 are the two hydrophobic pockets, and P3 is usually a polar region in the interface between /-tubulin. The physique was generated using PyMol (http://pymol.souceforge.net/). 3. Experimental Section 3.1. Chemistry The melting point was determined on a XT4A microscope melting-point apparatus (Keyi Electron Optical Instrument Factory, Beijing, China) without correction.1H NMR and 13C NMR spectra were recorded on BRUKER AVANCE 300 and 600 spectrometers (Bruker Company, Rheinstetten, Germany), with TMS as an internal standard and CDCl3 as the solvent. ESI mass spectra were performed on an API-3000 LC-MS spectrometer (Applied Biosystems, Toronto, ON, Canada). Flash column chromatography was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical, Qingdao, China). All solvents and reagents were purchased from commercial suppliers and, when necessary, were purified and dried by standard protocols. Organic solutions were dried over anhydrous sodium sulfate. The purity of the final compounds was assessed with an Agilent 1200 HPLC (Agilent Technologies, Santa Clara, CA, USA), and the results were greater than 95%. (= 1.5, 4.8 Hz), 8.75 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 4bCompact disc had been like the synthesis of intermadiate 4a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propanenitrile (5a) An assortment of 4a (0.62 g, 2.1 mmol), NaBH4 (0.32 g, 8.5 mmol) and 20 mL MeOH was heated under 50 C for 0.5 h. The blend was evaporated as well as the residue diluted with 25 mL EtOAc. The organic coating was dried out and filtered as well as the solvent eliminated by evaporation. Following the remedy was cooled and remained overnight to provide the substance 5a (2.18 g, 86.21%) like a white crystals. mp 129C130 C. 1H NMR (300 MHz, CDCl3): 3.10C3.24 (m, 2H), 3.82 (s, 6H), 3.85 (s, 3H), 3.99 (t, 1H), 6.40 (s, 2H), 7.09 (d, 2H, = 4.5 Hz), 8.56 (d, 2H, = 4.5 Hz). The artificial options for the intermediates 5bCompact disc had been like the synthesis of intermadiate 5a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propan-1-amine (6a) BF3O(C2H5)2 (7.5 mmol) was slowly put into a stirred solution of 5a (0.76 g, 2.5 mmol) and NaBH4 (10 mmol) in THF (10 mL) at 0 C. The perfect solution is was refluxed for 1 h, poured into drinking water, and extracted with EtOAc (15 mL 3). The mixed extracts had been dried out over anhydrous Na2SO4 and filtered. The solvents had been eliminated by evaporation to cover 6a (0.72 g, 96.61%) like a yellow essential oil. 1H NMR (300 MHz, CDCl3): 2.96C2.79 (m, 5H), 3.81 (s, 6H), 3.83 (s, 3H), 6.32 (s, 2H), 6.98 (dd, 2H, = 1.5, 4.5 Hz), 8.43 (dd, 2H, = 1.5, 4.5 Hz). SR 11302 The artificial options for the intermediates 6bCompact disc had been like the synthesis of intermadiate 6a. 3,4-Dimethoxy-= 1.5, 4.2 Hz). The artificial options for the intermediates 8C19 had been like the synthesis of intermadiate 7. 1-(3,4-Dimethoxyphenyl)-6,7,8-trimethoxy-4-(pyridin-4-ylmethyl)-3,4-dihydroisoquinoline (20) An assortment of 7 (0.70 g, 1.5 mmol), POCl3 (0.82 mL, 9 mmol) and CH3CN (15 mL) was stirred and heated under reflux for 4 h, then your solvents were removed by evaporation, as well as the residue was dissolved in EtOAc (30 mL). Then your remedy was neutralied to pH = 7 with saturated aqueous Na2CO3 and cleaned by drinking water (30 mL 3). The organic coating was dried out over anhydrous MgSO4 and filtered. Then your filtrate was focused under decreased pressure, The residue after evaporation was purified by adobe flash chromatography on silica gel (eluent: CH2Cl2/MeOH = 100:1 = 4.8, 14.7 Hz), 3.76 (s, 3H), 3.80 (s, 3H), 3.93 (s, 3H), 3.94 (s, 3H), 4.04 (dd, 1H, = 3.0, 14.4 Hz), 6.20 (s, 1H), 6.88 (d, 1H, = 8.4 Hz), 7.02 (dd, 1H, = 1.8, 5.1 Hz), 7.07 (dd, 2H, = 1.5, 4.5 Hz), 7.17 (d, 1H, = 1.8 Hz), 8.51 (dd, 2H, = 1.5, 4.5.21172260, 30901859, 21102174), Shanghai Organic Technology Foundation (Give No. substances is beneficial to activity. Open up in another window Shape 4 The hypothetical binding setting from the isomer of substance 32 to tubulin proteins. P1 and P2 will be the two hydrophobic wallets, and P3 can be a polar area in the user interface between /-tubulin. The shape was generated using PyMol (http://pymol.souceforge.net/). 3. Experimental Section 3.1. Chemistry The melting stage was determined on the XT4A microscope melting-point equipment (Keyi Electron Optical Device Manufacturer, Beijing, China) without modification.1H NMR and 13C NMR spectra were documented on BRUKER AVANCE 300 and 600 spectrometers (Bruker Business, Rheinstetten, Germany), with TMS as an interior standard and CDCl3 as the solvent. ESI mass spectra had been performed with an API-3000 LC-MS spectrometer (Applied Biosystems, Toronto, ON, Canada). Adobe flash column chromatography was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical substance, Qingdao, China). All solvents and reagents had been purchased from industrial suppliers and, when required, had been purified and dried out by regular protocols. Organic solutions had been dried out over anhydrous sodium sulfate. The purity of the ultimate substances was evaluated with an Agilent 1200 HPLC (Agilent Systems, Santa Clara, CA, USA), as well as the outcomes had been higher than 95%. (= 1.5, 4.8 Hz), 8.75 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 4bCompact disc had been like the synthesis of intermadiate 4a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propanenitrile (5a) An assortment of 4a (0.62 g, 2.1 mmol), NaBH4 (0.32 g, 8.5 mmol) and 20 mL MeOH was heated under 50 C for 0.5 h. The blend was evaporated as well as the residue diluted with 25 mL EtOAc. The organic coating was dried out and filtered as well as the solvent eliminated by evaporation. Following the remedy was cooled and remained overnight to provide the substance 5a (2.18 g, 86.21%) like a white crystals. mp 129C130 C. 1H NMR (300 MHz, CDCl3): 3.10C3.24 (m, 2H), 3.82 (s, 6H), 3.85 (s, 3H), 3.99 (t, 1H), 6.40 (s, 2H), 7.09 (d, 2H, = 4.5 Hz), 8.56 (d, 2H, = 4.5 Hz). The artificial options for the intermediates 5bCompact disc had been like the synthesis of intermadiate 5a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propan-1-amine (6a) BF3O(C2H5)2 (7.5 mmol) was slowly put into a stirred solution of 5a (0.76 g, 2.5 mmol) and NaBH4 (10 mmol) in THF (10 mL) at 0 C. The perfect solution is was refluxed for 1 h, poured into drinking water, and extracted with EtOAc (15 mL 3). The mixed extracts had been dried out over anhydrous Na2SO4 and filtered. The solvents had been eliminated by evaporation to cover 6a (0.72 g, 96.61%) like a yellow essential oil. 1H NMR (300 MHz, CDCl3): 2.96C2.79 (m, 5H), 3.81 (s, 6H), 3.83 (s, 3H), 6.32 (s, 2H), 6.98 (dd, 2H, = 1.5, 4.5 Hz), 8.43 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 6bCompact disc had been like the synthesis of intermadiate 6a. 3,4-Dimethoxy-= 1.5, 4.2 Hz). The artificial options for the intermediates 8C19 had been like the synthesis of intermadiate 7. 1-(3,4-Dimethoxyphenyl)-6,7,8-trimethoxy-4-(pyridin-4-ylmethyl)-3,4-dihydroisoquinoline (20) An assortment of 7 (0.70 g, 1.5 mmol), POCl3 (0.82 mL, 9 mmol) and CH3CN (15 mL) was stirred and heated under reflux for 4 h, then your solvents were removed by evaporation, as well as the residue was dissolved in EtOAc (30 mL). Then your remedy was neutralied to pH = 7 with saturated aqueous Na2CO3 and cleaned by drinking water (30 mL 3). The organic coating was dried out over anhydrous MgSO4 and filtered. Then your filtrate was focused under decreased pressure, The residue after evaporation was purified by adobe flash chromatography on silica gel (eluent: CH2Cl2/MeOH = 100:1 = 4.8, 14.7 Hz), 3.76 (s, 3H), 3.80 (s, 3H), 3.93 (s, 3H), 3.94 (s, 3H), 4.04 (dd, 1H, = 3.0, 14.4 Hz), 6.20 (s, 1H), 6.88 (d, 1H, = 8.4 Hz), 7.02 (dd, 1H, = 1.8, 5.1 Hz), 7.07 (dd, 2H, = 1.5, 4.5 Hz), 7.17 (d, 1H, = 1.8 Hz), 8.51 (dd, 2H, = 1.5, 4.5 Hz). ESI-MS (= 2.4, 14.7 Hz), 6.18 (s, 1H), 6.79 (d, 1H, =.The strongest compound 32 was confirmed to have the ability to inhibit tubulin polymerization and its own hypothetical binding mode to tubulin was proposed by molecular docking. amino group in the versatile sidechain of Lys352, which precisely clarify why the intro of 4′-nitro benzyl towards the C-4 placement from the isoquinoline band from the business lead substances is beneficial to activity. Open up in another window Shape 4 The hypothetical binding setting from the isomer of substance 32 to tubulin proteins. P1 and P2 will be the two hydrophobic wallets, and P3 can be a polar area in the user interface between /-tubulin. The shape was generated using PyMol (http://pymol.souceforge.net/). 3. Experimental Section 3.1. Chemistry The melting stage was determined on the XT4A microscope melting-point equipment (Keyi Electron Optical Device Manufacturer, Beijing, China) without modification.1H NMR and 13C NMR spectra were documented on BRUKER AVANCE 300 and 600 spectrometers (Bruker Business, Rheinstetten, Germany), with TMS as an interior standard and CDCl3 as the solvent. ESI mass spectra had been performed with an API-3000 LC-MS spectrometer (Applied Biosystems, Toronto, ON, Canada). Adobe flash column chromatography was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical substance, Qingdao, China). All solvents and reagents had been purchased from industrial suppliers and, when required, had been purified and dried out by regular protocols. Organic solutions had been dried out over anhydrous sodium sulfate. The purity of the ultimate substances was evaluated with an Agilent 1200 HPLC (Agilent Technology, SR 11302 Santa Clara, CA, USA), as well as the outcomes had been higher than 95%. (= 1.5, 4.8 Hz), 8.75 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 4bCompact disc had been like the synthesis of intermadiate 4a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propanenitrile (5a) An assortment of 4a (0.62 g, 2.1 mmol), NaBH4 (0.32 g, 8.5 mmol) and 20 mL MeOH was heated under 50 C for 0.5 h. The mix was evaporated as well as the residue diluted with 25 mL EtOAc. The organic level was dried out and filtered as well as the solvent taken out by evaporation. Following the alternative was cooled and remained overnight to provide the substance 5a (2.18 g, 86.21%) being a white crystals. mp 129C130 C. SR 11302 1H NMR (300 MHz, CDCl3): 3.10C3.24 (m, 2H), 3.82 (s, 6H), 3.85 (s, 3H), 3.99 (t, 1H), 6.40 (s, 2H), 7.09 (d, 2H, = 4.5 Hz), 8.56 (d, 2H, = 4.5 Hz). The artificial options for the intermediates 5bCompact disc had been like the synthesis of intermadiate 5a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propan-1-amine (6a) BF3O(C2H5)2 (7.5 mmol) was slowly put into a stirred solution of 5a (0.76 g, 2.5 mmol) and NaBH4 (10 mmol) in THF (10 mL) at 0 C. The answer was refluxed for 1 h, poured into drinking water, and extracted with EtOAc (15 mL 3). The mixed extracts had been dried out over anhydrous Na2SO4 and filtered. The solvents had been taken out by evaporation to cover 6a (0.72 g, 96.61%) being a yellow essential oil. 1H NMR (300 MHz, CDCl3): 2.96C2.79 (m, 5H), 3.81 (s, 6H), 3.83 (s, 3H), 6.32 (s, 2H), 6.98 (dd, 2H, = 1.5, 4.5 Hz), 8.43 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 6bCompact disc had been like the synthesis of intermadiate 6a. 3,4-Dimethoxy-= 1.5, 4.2 Hz). The artificial options for the intermediates 8C19 had been like the synthesis of intermadiate 7. 1-(3,4-Dimethoxyphenyl)-6,7,8-trimethoxy-4-(pyridin-4-ylmethyl)-3,4-dihydroisoquinoline (20) An assortment of 7 (0.70 g, 1.5 mmol), POCl3 (0.82 mL, 9 mmol) and CH3CN (15 mL) was stirred and heated under reflux for 4 h, then your solvents were removed by evaporation, as well as the residue was dissolved in EtOAc (30 mL). Then your alternative was neutralied to pH = 7 with saturated aqueous Na2CO3 and cleaned by drinking water (30 mL 3). The organic level was dried out over anhydrous MgSO4 and filtered. Then your filtrate was focused under decreased pressure, The residue after evaporation was purified by display chromatography on silica gel (eluent: CH2Cl2/MeOH = 100:1 = 4.8, 14.7 Hz), 3.76 (s, 3H), 3.80 (s, 3H), 3.93 (s, 3H), 3.94 (s, 3H), 4.04 (dd, 1H, = 3.0, 14.4 Hz), 6.20 (s, 1H), 6.88 (d, 1H, = 8.4 Hz), 7.02 (dd, 1H, = 1.8, 5.1 Hz), 7.07 (dd, 2H, = 1.5, 4.5 Hz), 7.17 (d, 1H, =.mp 129C130 C. isomer of substance 32 to tubulin proteins. P1 and P2 will be the two hydrophobic storage compartments, and P3 is normally a polar area in the user interface between /-tubulin. The amount was generated using PyMol (http://pymol.souceforge.net/). 3. Experimental Section 3.1. Chemistry The melting stage was determined on the XT4A microscope melting-point equipment (Keyi Electron Optical Device Stock, Beijing, China) without modification.1H NMR and 13C NMR spectra were documented on BRUKER AVANCE 300 and 600 spectrometers (Bruker Firm, Rheinstetten, Germany), with TMS as an interior standard and CDCl3 as the solvent. ESI mass spectra had been performed with an API-3000 LC-MS spectrometer (Applied Biosystems, Toronto, ON, Canada). Display column chromatography SR 11302 was performed with silica gel 300C400 mesh (Qingdao Haiyang Chemical substance, Qingdao, China). All solvents and reagents had been purchased from industrial suppliers and, when required, had been purified and dried out by regular protocols. Organic solutions had been dried out over anhydrous sodium sulfate. The purity of the ultimate substances was evaluated with an Agilent 1200 HPLC (Agilent Technology, Santa Clara, CA, USA), as well as the outcomes had been higher than 95%. (= 1.5, 4.8 Hz), 8.75 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 4bCompact disc had been like the synthesis of intermadiate 4a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propanenitrile (5a) An assortment of 4a (0.62 g, 2.1 mmol), NaBH4 (0.32 g, 8.5 mmol) and 20 mL MeOH was heated under 50 C for 0.5 h. The mix was evaporated as well as the residue diluted with 25 mL EtOAc. The organic level was dried out and filtered as well as the solvent taken out by evaporation. Following the alternative was cooled and remained overnight to provide the substance 5a (2.18 g, 86.21%) being a white crystals. mp 129C130 C. 1H NMR (300 MHz, CDCl3): 3.10C3.24 (m, 2H), 3.82 (s, 6H), 3.85 (s, 3H), 3.99 (t, 1H), 6.40 (s, 2H), 7.09 (d, 2H, = 4.5 Hz), 8.56 (d, 2H, = 4.5 Hz). The artificial options for the intermediates 5bCompact disc had been like the synthesis of intermadiate 5a. 3-(Pyridin-4-yl)-2-(3,4,5-trimethoxyphenyl)propan-1-amine (6a) BF3O(C2H5)2 (7.5 mmol) was slowly put into a stirred solution of 5a (0.76 g, 2.5 mmol) and NaBH4 (10 mmol) in THF (10 mL) at 0 C. The answer was refluxed for 1 h, poured into drinking water, and extracted with EtOAc (15 mL 3). The mixed extracts had been dried out over anhydrous Na2SO4 and filtered. The solvents had been taken out by evaporation to cover 6a (0.72 g, 96.61%) being a yellow essential oil. 1H NMR (300 MHz, CDCl3): 2.96C2.79 (m, 5H), 3.81 (s, 6H), 3.83 (s, 3H), 6.32 (s, 2H), 6.98 (dd, 2H, = 1.5, 4.5 Hz), 8.43 (dd, 2H, = 1.5, 4.5 Hz). The artificial options for the intermediates 6bCompact disc had been like the synthesis of intermadiate 6a. 3,4-Dimethoxy-= 1.5, 4.2 Hz). The artificial options for the intermediates 8C19 had been like the synthesis of intermadiate 7. 1-(3,4-Dimethoxyphenyl)-6,7,8-trimethoxy-4-(pyridin-4-ylmethyl)-3,4-dihydroisoquinoline (20) An assortment of 7 (0.70 g, 1.5 mmol), POCl3 (0.82 mL, 9 mmol) and CH3CN (15 mL) was stirred and heated under reflux for 4 h, then your solvents were removed by evaporation, as well as the residue was dissolved in EtOAc (30 mL). Then your option was neutralied to pH = 7 with saturated aqueous Na2CO3 and cleaned by drinking water (30 mL 3). The organic level was dried out over anhydrous MgSO4 and filtered. Then your filtrate was focused under decreased pressure, The residue after evaporation was purified by display chromatography on silica gel (eluent: CH2Cl2/MeOH = 100:1 = 4.8, 14.7 Hz), 3.76 (s, 3H), 3.80 (s, 3H), 3.93 (s, 3H), 3.94 (s, 3H), 4.04 (dd, 1H, = 3.0, 14.4 Hz), 6.20 (s, 1H), 6.88 (d, 1H, = 8.4 Hz), 7.02 (dd, 1H, =.