Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 Electronic Supporting Information Novel 1-(1-benzyl-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodides are competitive antagonists of the human 4 2 and 7 nicotinic acetylcholine receptors Edwin G. Pérez, Cristian Ocampo, Dominik Feuerbach, Jhon J. López, Ricardo Tapia, and Hugo R. Arias General Experimental . . . . . . . . ………………………………………………………….. 1 General methods………………………………………………………………. . . . . . . . . 1 Calcium influx and radioligand binding experiments………………………….. . . . . . . . 1 Homology Modeling ………………………………………………………………………..2 Molecular Docking ………………………………………………………………………..2 References………………………………………………………………………………….2 Synthetic Procedures and NMR description………………………………………… . . . . 4 1 H and 13C NMR Spectra of Compounds . . . …………………………………………. . ..36 General Experimental General methods Calcium influx and radioligand binding experiments. The concentration–response data for the Ca2+ influx and radioligand binding experiments were curve-fitted, and the IC50 and Hill coefficient (nH) values were calculated by nonlinear 1 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 least squares analysis using the Prism software (GraphPad Software, San Diego, CA), based on the following equation: = 1 / [l + ([L] / IC50)nH] (1) where is the fractional amount of the radioligand bound in the presence of inhibitor at a concentration [L] compared to the amount of the radioligand bound in the absence of inhibitor (total binding). The observed IC50 values from the competition experiments described above were transformed into inhibition constant (Ki) values using the Cheng– Prusoff relationship: Ki = IC50 / {1 + ([[3H]ligand] / Kdligand)} (2) where [[3H]ligand] is the initial concentration of [3H]MLA, [3H]cytisine, or [3H]imipramine, respectively, and Kdligand is the dissociation constant for [3H]MLA at the h 7 nAChR (1.86 nM)2, for [3H]cytisine at the h 4 2 nAChR (0.3 nM)3, and for [3H]imipramine at the h 4 2 (0.83 µM)4 and h 7 (1 µM)5 nAChRs, respectively. Homology Modeling To construct the extracellular domain of the h 7 and h 4 2 nAChRs, the structure of the acetylcholine binding protein from Lymnaea stagnalis (Ls-AChBP) in complex with nicotine at 2.2 Å resolution (PDB 1UW6)6 was used as a template for homology modeling. The target protein and template were aligned using the Multalin server.7 The models were built using the MODELLER9v6.8 In this study, 100 runs were carried out using standard parameters and the outcomes were ranked on the basis of the internal scoring function of the program. The best model was chosen as the target model. Molecular Docking In order to obtain information about the most important nAChR-ligand interactions, molecular docking experiments were performed using AutoDock 4.0.9 For the docking to the orthosteric binding sites, the grid maps were calculated using the autogrid option and centered on the binding sites. The volumes chosen for the grid maps were made up of 60 × 60 × 60 points, with a grid-point spacing of 0.375 Å. The autotors option was used to define the rotating bonds in the ligand. In the Lamarckian genetic algorithm dockings, the number of individuals in a population of 1,500, a maximum number of 2.5 × 106 energy evaluations, a maximum number of 27,000 generations, a mutation rate of 0.02, and a cross-over rate of 0.80, were employed. The ligands 4b, and 4c docked to the respective nAChR were built using the lowest docked-energy binding positions using Gaussian03,10 and the partial charges were corrected using electrostatic potential methodology.11, 12 The energy of binding of the best docked compounds was calculated by AutoDock 4.0, AutoDock Vina, and by molecular mechanics (i.e., difference between the energy of the ligand-receptor complex and the sum of energies of the isolated receptor and ligand). References 1. Cheng, Y.-C.; Prusoff, W. H. Biochem. Pharmacol. 1973, 22, 3099 2. Davies, A. R.; Hardick, D. J.; Blagbrough, I. S.; Potter, B.V.; Wolstenholme, A. J.; Wonnacott, S. Neuropharmacology 1999, 38, 679. 3. Zhang, J; Steinbach, J. H. Brain Res. 2003, 959, 98. 2 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 4. Arias, H. R.; Rosenberg, A.; Targowska-Duda, K. M.; Feuerbach, D.; Jozwiak, K.; Moaddel, R.; Weiner, I. W. Int. J. Biochem. Cell Biol. 2010, 42, 1007. 5. Arias, H. R.; Feuerbach, D.; Targowska-Duda K, M.; Russell, M. M.; Jozwiak, K. Biochemistry 2010, 49, 5734. 6. Celie PHN, Van Rossum-Fikkert SE, Van Dijk WJ, Brejc K, Smit AB, Sixma TK. Nicotine and carbamylcholine binding to nicotinic acetylcholine receptors as studied in AChBP crystal structures. Neuron 2004;41:907-14. 7. Corpet F. Multiple sequence alignment with hierarchical clustering. Nucl Acids Res 1988;16:10881-90. 8. Šali A, Blundell TL. Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 1993;234:779- 815. 9. Morris GM, Goodsell DS, Halliday RS, Huey R, Hart WE, Belew RK, Olson AJ. Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 1998;19:1639-62. 10. Frisch M J, Trucks G W, Schlegel H B, Scuseria, G E, Robb M A, Cheeseman J R, Montgomery Jr J.A, Vreven T, Kudin K N, Burant J C, Millam J M, Iyengar S S, Tomasi J, Barone V, Mennucci B, Cossi M, Scalmani G, Rega N, Petersson G A, Nakatsuji H, Hada M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao O, Nakai H, Klene M, Li X, Knox J E, Hratchian H P, Cross J B, Bakken V, Adamo C, Jaramillo J, Gomperts R, Stratmann R E, Yazyev O, Austin A J, Cammi R, Pomelli C, Ochterski J W, Ayala P Y, Morokuma K, Voth G A, Salvador P, Dannenberg J J, Zakrzewski VG, Dapprich S, Daniels A D, Strain M C, Farkas O, Malick D K, Rabuck A D, Raghavachari K, Foresman J B, Ortiz J V, Cui Q, Baboul A G, Clifford S, Cioslowski J, Stefanov B B, Liu G, Liashenko A, Piskorz P, Komaromi I, Martin R L, Fox D J, Keith T, Al-Laham M A, Peng C Y, Nanayakkara A, Challacombe M, Gill P M W, Johnson B, Chen W, Wong M W, Gonzalez C, and Pople J A. Gaussian, Inc., Wallingford CT, 2004. 11. Chipot C, Maigret B, Rivail J-L, Sheraga HA. Modeling amino-acid side-chains. 1. Determination of net atomic charges from abinitio self-consistent-field molecular electrostatic properties. J Phys Chem 1992;96:10276-84. 12. Ángyán JG. Choosing between alternative MP2 algorithms in the self-consistent reaction field theory of solvent effects. Chem Phys Lett 1995;241:51-6. 3 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 Synthetic Procedures Microwave reactions were carried out in a CEM Discover reactor. Melting points were determined on a Reichert Galen III hot plate microscope apparatus and are uncorrected. 1H NMR spectra were recorded on Bruker AMX 200, Bruker Avance 400, instruments at 200 or 400 MHz, respectively. 13 C NMR were recorded on the same instruments at 50, or 100 MHz. Chemical shifts are reported in values (part per million, ppm) relative to an internal standard of tetramethylsilane in CDCl3 or DMSO-d6 and coupling constants (J) are given in Hertz. The elemental analyses were performed in a Fison SA, model EA-1108 apparatus. Precoated silica gel 60 plates (Merck 60 F254 0.2 mm) were used for TLC. TLC spots were visualized by spraying with Dragendorff's reagent, by exposing to iodine vapor or UV light. General Procedure for microwave reaction: A 50 mL microwave reaction tube was charged with aldehyde (5 mmol, 1 equiv.), indoline (6.0 mmol, 1.2 equiv.), toluene (10 mL), and benzoic acid (1.0 mmol, 0.2 equiv.). The reaction tube was sealed and heated in the microwave reactor at 200 ºC, (200 W, 50–170 psi) for the appropriate time. After cooling with compressed air flow, the crude reaction mixture was diluted with EtOAc (25 mL) and washed with saturated aqueous NaHCO3 (3 x 25 mL). The aqueous layers were extracted with EtOAc (3 x 25 mL) and the combined organic layers dried over anhydrous Na2SO4. The solvent was removed in vacuo and the product was purified by silica gel column chromatography (hexane:EtOAc). 4 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1-Bencyl-1H-indole (3a) Following the general procedure, compound 3a was obtained from indoline and benzaldehyde (2a) as colorless liquid in 27% yield (column chromatography Hexanes: EtOAc 9:1). 1 H NMR (200 MHz, CDCl3): 7.82 – 7.65 (m, 1H), 7.46 – 7.29 (m, 4H), 7.28 – 7.22 (m, 1H), 7.22 – 7.10 (m, 4H), 6.64 (dd, J = 3.2, 0.8 Hz, 1H), 5.35 (s, 2H). 13C NMR (50 MHz, CDCl3): 137.5 (C-9), 128.7 (C-4´), 128.7 (C-6´), 128.2 (C-2), 127.6 (C-5´), 127.6 (C-4), 126.7 (C-7´), 126.7 (C-3´), 121.7 (C-5), 121.0 (C-6), 119.5 (C-7), 109.7 (C-8), 101.6 (C-3), 50.0 (C-1´). 5 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 3-((1H-Indol-1-yl)methyl)phenol (3b). Following the general procedure, compound 3b was obtained from indoline and 3hydroxybenzaldehyde (2b) as colorless liquid in 21% yield (column chromatography Hexanes: EtOAc 7:3). 1 H NMR (200 MHz, (CD3)2C=0 ): 7.67 (d, J = 7.3 Hz, 1H), 7.52 – 7.38 (m, 2H), 7.17 (ddd, J = 18.9, 11.4, 5.6 Hz, 3H), 6.79 (t, J = 7.0 Hz, 2H), 6.68 – 6.55 (m, 2H), 5.43 (s, 2H), 3.16 (d, J = 11.4 Hz, 1H). 13C NMR (50 MHz, (CD3)C=0): 158.5 (C-4´), 140.8 (C- 2´), 137.1 (C-9), 130.4 (C-6´), 129.7 (C-2), 129.5 (C-4), 128.8 (C-5), 121.4 (C-6), 119.9 (C-7´), 118.7 (C-7), 115.1 (C-3´), 114.3 (C-5´), 110.7 (C-8), 101.9 (C-3), 50.1 (C-1´´). 6 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 4-((1-H-indol-1-yl)methyl)phenol (3c). Following the general procedure, compound 3c was obtained from indoline and 4hydroxybenzaldehyde (2c) as colorless solid in 17% yield (column chromatography Hexanes: EtOAc 7:3). Mp. 58,0-60,0 °C. 1 H NMR (200 MHz, CDCl3): 7.64 (d, J = 7.1 Hz, 1H), 7.27 (d, J = 7.7 Hz, 1H), 7.10 (dd, J = 14.7, 5.2 Hz, 3H), 6.94 (d, J = 8.3 Hz, 2H), 6.64 (d, J = 8.4 Hz, 2H), 6.51 (d, J = 3.0 Hz, 1H), 5.17 (s, 2H). 13C NMR (50 MHz, CDCl3): 155.0 (C-5´), 136.2 (C-9), 129.6 (C- 4), 128.7 (C-2´), 128.4 (C-7´), 128.4 (C-3´), 128.2 (C-2), 121.6 (C-5), 121.0 (C-6), 119.5 (C-7), 115.6 (C-6´), 115.6 (C-4´), 109.8 (C-8), 101.5 (C-3), 49.6 (C-1´). 7 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1-(4-Methoxybencyl)-1H-indole (3d). Following the general procedure, compound 3d was obtained from indoline and 4methoxybenzaldehyde (2d) as colorless liquid in 21% yield (column chromatography Hexanes: EtOAc 8:2). 1 H NMR (200 MHz, CDCl3): 7.73 – 7.50 (m, 1H), 7.27 (d, J = 2.6 Hz, 1H), 7.21 – 7.09 (m, 2H), 7.08 – 7.03 (m, 2H), 7.01 (d, J = 2.8 Hz, 1H), 6.92 – 6.69 (m, 2H), 6.52 (dd, J = 3.2, 0.8 Hz, 1H), 5.21 (s, 2H), 3.73 (s, 3H). 13 C NMR (50 MHz, CDCl3): 159.1 (C-5´), 136.2 (C-9), 129.5 (C-2), 128.7 (C-4), 128.2 (C-7´), 128.2 (C-3´), 128.1 (C-2´),128.1 (C-5), 121.6 (C-5), 121.0 (C-6), 119.5 (C-7), 114.1 (C-6´), 114.1 (C-4´), 109.7 (C-8), 101.5 (C-3), 55.3 (C-8´), 49.6 (C-1´). 8 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 5-((1H-Indol-1-yl)methyl)-2-methoxyphenol (3e). Following the general procedure, compound 3e was obtained from indoline and 3-hydroxy4-methoxybenzaldehyde (2e) as colorless liquid in 35% yield (column chromatography dichloromethane). 1 H NMR (200 MHz, CDCl3): 7.67 – 7.56 (m, 1H), 7.32 – 7.22 (m, 1H), 7.21 – 7.11 (m, 1H), 7.12 – 7.04 (m, 2H), 6.76 – 6.66 (m, 2H), 6.62 – 6.53 (m, 1H), 6.50 (dd, J = 3.2, 0.9 Hz, 1H), 5.58 (s, 1H), 5.17 (s, 2H), 3.78 (s, 3H). 13C NMR (50 MHz, CDCl3): 145.8 (C- 5´), 137.5 (C-4´), 130.8 (C-9), 128.7 (C-2), 128.2 (C-2´), 121.6 (C-4), 120.9 (C-5), 119.4 (C-6), 118.5 (C-7), 113.3 (C-7´), 110.7 (C-3´), 109.7 (C-6´), 101.5 (C-8), 56.0 (C-1´), 49.7 (C-8´). 9 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 4-((1H-Indol-1-yl)methyl)-2-methoxyphenol (3f). Following the general procedure, compound 3f was obtained from indoline and 4methoxybenzaldehyde (2f) as colorless liquid in 25% yield (column chromatography dichloromethane). 1 H NMR (200 MHz, CDCl3 ): 7.71 – 7.57 (m, 1H), 7.29 (d, J = 8.2 Hz, 1H), 7.24 – 7.14 (m, 1H), 7.14 – 7.04 (m, 2H), 6.83 (d, J = 8.1 Hz, 1H), 6.69 – 6.62 (m, 1H), 6.63 – 6.46 (m, 2H), 5.20 (s, 2H), 3.73 (s, 3H). 13C NMR (50 MHz, CDCl3): 146.8 (C-4´), 145.1 (C-5´), 136.3 (C-9), 129.3 (C-2), 128.7 (C-2´), 128.0 (C-4), 121.6 (C-7´), 120.9 (C-5), 120.1 (C-6), 119.5 (C-7), 114.4 (C-6´), 109.7 (C-3´), 109.5 (C-8), 101.5 (C-3), 55.9 (C-1´), 50.0 (C-8´). 10 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1-(3,4-dimethoxybencyl)-1H-indole (3g). Following the general procedure, compound 3g was obtained from indoline and 3,4dimethoxybenzaldehyde (2g) as pink solid in 52% yield (column chromatography Hexanes: EtOAc 8:2); m.p: 67.9-69.8 °C. 1 H NMR (200 MHz, CDCl3): 7.67 (m, 1H), 7.34 (m, 1H), 7.22 (d, J = 5.5 Hz, 1H), 7.16 (d, J = 1.8 Hz, 1H), 7.13 – 7.12 (m, 1H), 6.82 – 6.76 (m, 1H), 6.71 – 6.66 (m, 2H), 6.56 (dd, J = 3.2, 0.8 Hz, 1H), 5.26 (s, 2H), 3.85 (s, 3H), 3.79 (s, 3H). CDCl3): 13 C NMR (50 MHz, 149.2 (C-4´), 148.5 (C-5´), 136.3 (C-9), 129.9 (C-2´), 128.7 (C-2), 128.1 (C-4), 121.6 (C-7´), 120.9 (C-7), 119.5 (C-5), 119.3 (C-6), 111.2 (C-8), 110.1 (C-6´), 109.7 (C3´), 101.6 (C-3), 55.9 (C-1´), 49.9 (C-8´), 49.1 (C-9´). 11 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1-(Benzo[d][1,3]dioxol-4-ylmethyl)-1H-indole (3h). Following the general procedure, compound 3h was obtained from indoline and 3,4dimethoxybenzaldehyde (2h) as yellow solid in 41% yield (column chromatography Hexanes: EtOAc 8:2); m.p: 78.8-80.1 °C. 1 H NMR (200 MHz, CDCl3): 7.68 (d, J = 7.3 Hz, 1H), 7.32 (d, J = 7.8 Hz, 1H ), 7.28 – 7.19 (m, 1H), 7.18 – 7.08 (m, 2H), 6.76 (d, J = 7.8 Hz, 1H), 6.68 (s, 1H), 6.63 – 6.52 (m, 2H), 5.93 (s, 2H), 5.24 (s, 2H). 13C NMR (50 MHz, CDCl3): 148.1 (C-3´), 147.1 (C-4´), 136.2 (C-9), 131.3 (C-2), 128.7 (C-4), 128.1 (C-2´), 121.7 (C-6´), 121.0 (C-7), 120.2 (C7´), 119.5 (C-6), 109.6 (C-5), 108.3 (C-8), 107.4 (C-8´), 49.9 (C-1´). 12 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 4-((1H-Indol-1-yl)methyl)-N,N-dimethylaniline (3i). Following the general procedure, compound 3i was obtained from indoline and 4dimethylaminebenzaldehyde (2i) as colorless solid in 21% yield (column chromatography Hexanes: EtOAc 8:2). M.p: 107,2-108,8 °C. 1 H NMR (200 MHz, CDCl3): 7.61 (ddd, J = 7.3, 1.6, 0.8 Hz, 1H), 7.41 – 7.23 (m, 1H), 7.21 – 7.12 (m, 1H), 7.12 – 7.04 (m, 2H), 7.02 – 6.96 (m, 2H), 6.71 – 6.56 (m, 2H), 6.48 (dd, J = 3.2, 0.9 Hz, 1H), 5.18 (s, 2H), 2.88 (s, 6H). 13C NMR (50 MHz, CDCl3): 150.1 (C-5´), 136.3 (C-9), 128.7 (C-2), 128.1 (C-7´), 128.19 (C-3´), 128.11 (C-4), 125.3 (C-2´), 13 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 121.4 (C-5), 120.9 (C-6), 119.3 (C-7), 112.7 (C-6´), 112.7 (C-4´), 109.8 (C-8), 101.2 (C-3), 49.7 (C-1´), 40.6 (C-8´), 40.6 (C-9´). 1-(pyridin-3-ylmethyl)-1H-indole (3j). Following the general procedure, compound 3j was obtained from indoline and 3pyridinecarboxaldehyde (2j) as colorless solid in 59% yield (column chromatography Hexanes: EtOAc 8:2); m.p: 58,6-60,5 °C. 1 H NMR (200 MHz, CDCl3): 8.52 (s, 2H), 7.66 (d, J = 7.3 Hz, 1H), 7.35 – 7.22 (m, 2H), 7.14 (dd, J = 10.3, 3.6 Hz, 4H), 6.57 (d, J = 3.0 Hz, 1H), 5.32 (s, 2H). 13C NMR (50 MHz, CDCl3): 149.2 (C-3´), 148.3 (C-5´), 136.0 (C-9), 134.4 (C-7´), 133.1 (C-2´), 128.8 (C-4), 127.9 (C-2), 123.7 (C-6´), 122.0 (C-5), 121.2 (C-6), 119.8 (C-7), 109.4 (C-8), 102.3 (C-3), 47.6 (C-1´). 14 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 General procedure for the Mannich reaction. To a solution of N1-bencylindole derivative (3, 1 mmol) in acetic acid (3.35 ml) at room temperature was added 40% aqueous dimethylamine (6,85 mmol), formaldehyde (2,76 mmol) and was stirred for 24 h under a nitrogen atmosphere. The reaction was quenched with 10% KOH (pH = 10) and extracted with EtoAc (3 × 50 mL), the organic layer was dried over MgSO4 and filtered. The compounds were characterized and used in the next reaction without further purification. 1-(1-benzyl-1H-indol-3-yl)-N,N-dimethylmethanamine (3a-1). 15 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 Following the general procedure, compound 3a-1 was obtained from 3a as pink solid. Yield: 92%. M.p: 57,3-59,1 °C. 1 H NMR (200 MHz, CDCl3): 7.70 (d, J = 7.5 Hz 1H), 7.25 (s, 4H), 7.10 (d, J = 4.5 Hz, 5H), 5.29 (s, 2H), 3.66 (s, 2H), 2.29 (s, 6H). 13 C NMR (50 MHz, CDCl3): 137.5 (C-9), 136.5 (C-2´), 128.7 (C-6´), 128.7 (C-4´), 128.6 (C-4), 127.9 (C-5´), 127.6 (C-2), 126.8 (C7´), 126.8 (C-3’), 121.7 (C-5), 119.4 (C-6), 119.4 (C-7), 111.9 ( C-8), 109.7 (C-3), 54.2 (C1´´), 49.9 (C-1´), 45.1 (-NCH3)2. 3-((3-((dimethylamino)methyl)-1H-indol-1-yl)methyl)phenol (3b-1). Following the general procedure, compound 3b-1 was obtained from 3b as colorless liquid. Yield: 80%. 1 H NMR (200 MHz, (CD3)2C=0): 7.57 – 7.40 (m, 1H), 7.22 (s, 2H), 7.12 (s, 3H), 6.76 (dd, J = 9.8, 4.2 Hz, 2H), 6.70 – 6.45 (m, 2H), 5.41 (s, 2H), 4.00 (s, 2H), 3.51 (s, 1H), 2.50 16 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 (s, 6H). 13C NMR (50 MHz, (CD3)C=0): 158.9 (C-4´), 140.5 (C-2´), 130.5 (C-9), 130.2 (C-6´), 129.4 (C-2), 126.5 (C-4), 122.3 (C-5), 120.0 (C-6), 119.5 (C-7´), 118.2 (C-7), 115.3 (C-3´), 114.1 (C-5´), 110.9 (C-8), 107.2 (C-3), 53.8 (C-1´´), 50.1 (C-1´), 43.9 (-NCH3)2. 4-((3-((dimethylamino)methyl)-1H-indol-1-yl)methyl)phenol (3c-1). Following the general procedure, compound 3c-1 was obtained from 3c as white solid. Yield: 97%.m.p: 111,2-112,9 °C. 1 H NMR (200 MHz, (CD3)2C=O/CD3OD): 7.65 – 7.55 (m, 1H), 7.28 (dd, J = 6.7, 1.9 Hz, 1H), 7.23 (s, 1H), 7.16 – 7.02 (m, 2H), 6.91 (d, J = 8.6 Hz, 2H), 6.71 (d, J = 8.6 Hz, 2H), 5.11 (s, 2H), 3.93 (s, 2H), 2.43 (s, 6H). 13C NMR (50 MHz, (CD3)2C=O/CD3OD ): 157.8 (C-5´), 137.0 (C-9), 130.3 (C-4), 129.3 (C-2´), 129.1 (C-7´), 129.1 (C-3´), 128.3 (C- 17 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 2), 122.5 (C-5), 120.4 (C-6), 119.5 (C-7), 116.3 (C-6´), 116.3 (C-4´), 110.8 (C-8), 108.0 (C-3), 53.2 (C-1´), 50.3 (C-1´´), 43.8 (-NCH3)2. 1-(1-(4-methoxybenzyl)-1H-indol-3-yl)-N,N-dimethylmethanamine (3d-1). Following the general procedure, compound 3d-1 was obtained from 3d as colorless liquid. Yield: 98%. 1 H NMR (200 MHz, CDCl3): 7.75 – 7.66 (m, 1H), 7.31 – 7.23 (m, 1H), 7.23 – 7.15 (m, 1H), 7.14 – 7.10 (m, 1H), 7.09 – 7.06 (m, 1H), 7.05 (d, J = 0.6 Hz, 2H), 6.88 – 6.76 (m, 2H), 5.22 (s, 2H), 3.76 (s, 3H), 3.62 (s, 2H), 2.28 (s, 6H). 13C NMR (50 MHz, CDCl3): 159.0 (C-5´), 136.5 (C-9), 129.5 (C-2), 128.6 (C-4), 128.2 (C-7´), 128.2 (C-3´), 127.5 (C18 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 2´), 121.6 (C-5), 119.4 (C-6), 119.2 (C-7), 114.1 (C-6´), 114.1 (C-4´), 112.3 (C-8), 109.6 (C-3), 55.3 (C-8´), 54.4 (C-1´), 49.4 (C-1´´), 45.4 (-NCH3)2. 5-((3-((dimethylamino)methyl)-1H-indol-1-yl)methyl)-2-methoxyphenol (3e-1). Following the general procedure, compound 3e-1 was obtained from 3e as colorless liquid. Yield: 94%. 1 H NMR (200 MHz, CDCl3): 7.60 (dd, J = 6.3, 2.2 Hz, 1H), 7.29 – 7.19 (m, 1H), 7.17 – 7.07 (m, 2H), 7.02 (s, 1H), 6.74 (d, J = 8.2 Hz, 1H), 6.60 (dd, J = 8.2, 2.1 Hz, 1H), 6.46 (s, 1H), 5.13 (s, 2H), 3.81 (s, 3H), 3.50 (s, 2H), 2.21 (s, 6H). 13C NMR (50 MHz, CDCl3): 19 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 146.7 (C-5´), 146.6 (C-4´), 136.3 (C-9), 130.4 (C-2), 128.6 (C-2´), 128.2 (C-4), 121.7 (C5), 119.4 (C-6), 119.0 (C-7), 117.8 (C-7´), 113.4 (C-3´), 111.0 (C-6´), 110.4 (C-8), 109.8 (C-3), 55.9 (C-1´), 53.4 (C-1´´), 49.4 (C-8´), 44.4 (-NCH3)2. 4-((3-((dimethylamino)methyl)-1H-indol-1-yl)methyl)-2-methoxyphenol (3f-1). Following the general procedure, compound 3f-1 was obtained from 3f as colorless liquid. Yield: 97%. 1 H NMR (200 MHz, CDCl3 ): 7.58 (dd, J = 6.5, 2.6 Hz, 1H), 7.31 (s, 2H), 7.16 (dd, J = 7.2, 4.6 Hz, 2H), 6.80 (d, J = 8.5 Hz, 1H), 6.62 (s, 2H), 5.18 (s, 2H), 4.08 (s, 2H), 3.74 (s, 3H), 2.52 (s, 6H). 13C NMR (50 MHz, CDCl3): 20 147.2 (C-4´), 145.7 (C-5´), 136.3 (C-9), Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 130.3 (C-4), 128.5 (C-2´), 128.3 (C-2), 122.2 (C-7), 120.25 (C-7´), 120.21 (C-6), 118.4 (C5), 114.7 (C-6´), 110.2 (C-3´), 109.8 (C-8), 105.3 (C-3), 55.8 (C-1´), 51.8 (C-1´´), 50.2 (C8´), 42.3 (-NCH3)2. 1-(1-(3,4-dimethoxybenzyl)-1H-indol-3-yl)-N,N-dimethylmethanamine (3g-1). Following the general procedure, compound 3g-1 was obtained from 3g as colorless liquid. Yield: 92%. 1 H NMR (200 MHz, CDCl3): 7.70 (dd, J = 6.4, 1.9 Hz, 1H), 7.31 (dd, J = 7.1, 2.2 Hz, 1H), 7.24 – 7.16 (m, 1H), 7.13 (d, J = 1.5 Hz, 1H), 6.79 (d, J = 8.5 Hz, 1H), 6.74 – 6.63 (m, 2H), 5.24 (s, 2H), 3.88 (s, 3H), 3.78 (s, 3H), 3.73 (s, 2H), 2.33 (s, 6H). 13C NMR (50 MHz, CDCl3): 149.2 (C-4´), 148.5 (C-5´), 136.5 (C-9), 129.8 (C-2´), 128.6 (C-4), 128.0 (C-2), 21 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 121.7 (C-7´), 119.4 (C-7), 119.37 (C-6), 119.3 (C-5), 111.2 (C-6´), 111.0 (C-3), 110.1 (C8), 109.7 (C-3´), 55.9 (C-1´), 55.8 (C-1´´), 49.8 (-NCH3)2, 44.6 (C-8´), 44.6 (C-9´). 1-(1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-indol-3-yl)-N,N-dimethylmethanamine (3h-1). Following the general procedure, compound 3h-1 was obtained from 3h as colorless liquid. Yield: 95%. 1 H NMR (200 MHz, CDCl3): 7.77 – 7.67 (m, 1H), 7.29 (dd, J = 6.8, 1.8 Hz, 1H), 7.24 – 7.11 (m, 2H), 7.09 (d, J = 2.4 Hz, 1H), 6.71 (dd, J = 14.8, 4.4 Hz, 1H), 5.92 (s, 2H), 5.20 (s, 2H), 3.67 (s, 2H), 2.31 (s, 6H). 13C NMR (50 MHz, CDCl3): 22 148.0 (C-6´), 147.1 (C-5´), Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 136.4 (C-9), 131.3 (C-2´), 128.6 (C-4), 127.6 (C-2), 121.7 (C-3´), 120.2 (C-7), 119.4 (C-5), 119.4 (C-6), 112.0 (C-3), 109.6 (C-4´), 108.3 (C-7´), 107.5 (C-8), 101.1 (C-8´), 54.2 (C1´´), 49.8 (C-1´), 45.1 (-NCH3)2. 4-((3-((dimethylamino)methyl)-1H-indol-1-yl)methyl)-N,N-dimethylaniline (3i-1). Following the general procedure, compound 3i-1 was obtained from 3i as white solid. Yield: 98%. M.p: 98-100 °C 1 H NMR (200 MHz, CDCl3): 7.68 (dd, J = 6.7, 1.7 Hz, 1H), 7.31 (dd, J = 7.0, 1.5 Hz, 1H), 7.14 (ddd, J = 14.5, 7.0, 1.4 Hz, 2H), 7.07 – 7.00 (m, 3H), 6.70 – 6.58 (m, 2H), 5.17 (s, 2H), 3.60 (s, 2H), 2.90 (s, 6H), 2.26 (s, 6H). 13C NMR (50 MHz, CDCl3): 23 150.1 (C- Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 5´), 136.5 (C-9), 128.6 (C-4), 128.2 (C-7´), 128.2 (C-3´), 127.6 (C-2), 125.0 (C-2´), 121.5 (C-5), 119.3 (C-6), 119.1 (C-7), 112.6 (C-6´), 112.6 (C-4´), 111.9 (C-8), 109.7 (C-3), 54.5 (C-1´´), 49.5 (C-1´), 45.3 (-NCH3)2, 40.6 (C-8´), 40.6 (C-9´). N,N-dimethyl-1-(1-(pyridin-3-ylmethyl)-1H-indol-3-yl)methanamine (3j-1). Following the general procedure, compound 3j-1 was obtained from 3j as colorless liquid. Yield: 97%. 1 H NMR (200 MHz, CDCl3): 8.49 (d, J = 2.9 Hz, 2H), 7.68 (ddd, J = 6.4, 3.1, 2.4 Hz, 1H), 7.29 (ddd, J = 7.9, 5.1, 1.9 Hz, 1H), 7.22 – 7.18 (m, 1H), 7.17 – 7.13 (m, 2H), 7.10 24 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 (dd, J = 5.4, 3.1 Hz, 2H), 5.29 (s, 2H), 3.67 (s, 2H), 2.30 (s, 6H). CDCl3): 13 C NMR (50 MHz, 149.2 (C-3´), 148.3 (C-5´), 136.3 (C-9), 134.4 (C-7´), 133.0 (C-2´), 128.7 (C-4), 127.7 (C-2), 123.7 (C-6´), 122.1 (C-5), 119.7 (C-6), 119.5 (C-7), 112.2 (C-8), 109.4 (C-3), 53.9 (C-1´´), 47.5 (C-1´), 44.9 (-NCH3)2. General procedure for synthesis of N,N,N-trimethylammonium derivarives. To a solution of gramine (3, 0.5 mmol) in acetone (5 mL), iodomethane (8.0 mmol) was added and the reaction mixture was stirred for 16 h. Then, Et2O (20 mL) was added and the solid collected by filtration to afford the desired trimethylammonium compound. 4a-j. 1-(1-benzyl-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4a). 25 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 Following the general procedure, compound 4a was obtained from 3a-1 as white solid. Yield: 96%. M. p: 167,7-172,1 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.90 (d, J = 3.7 Hz, 2H), 7.54 (d, J= 7.3 Hz, 1H), 7.38 – 7.29 (m, 2H), 7.26 (t, J = 6.6 Hz, 3H), 7.18 (dd, J = 13.1, 6.7 Hz, 2H), 5.53 (s, 2H), 4.75 (s, 2H), 3.08 (s, 9H). 13C NMR (100 MHz, DMSO-d6): 137.9 (C-9), 136.3 (C-2´), 133.9 (C- 4), 129.0 (C-6´), 129.0 (C-4´), 128.7 (C-2), 127.9 (C-5´), 127.4 (C-7´), 127.4 (C-3´), 122.5 (C-7), 120.8 (C-6), 119.3 (C-5), 111.3 (C-8), 102.0 (C-3), 60.6 (C-1´´), 51.6 (-NCH3)3, 49.7 (C-1´). Anal. calculated for: C, 56.17; H, 5.71; N, 6.89, Found: C,56.05; H,5.67; N,6.95 1-(1-(3-hydroxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4b). 26 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 Following the general procedure, compound 4b was obtained from 3b-1 as white solid. Yield: 98%. M. p: 151,1-156,8 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.94 – 7.82 (m, 2H), 7.51 (d, J = 7.8 Hz, 1H), 7.16 (ddd, J = 21.3, 14.8, 7.2 Hz, 2H), 6.67 (d, J = 8.9 Hz, 1H), 6.53 (s, 2H), 5.44 (s, 1H), 4.75 (s, 2H), 3.13 (d, J= 10.7 Hz, 1H), 3.08 (s, 9H). 13C NMR (100 MHz, DMSO-d6): 157.9 (C- 4´), 139.4 (C-2´), 136.3 (C-9), 134.0 (C-6´), 130.0 (C-2), 128.6 (C-4), 122.4 (C-5), 120.8 (C-6), 119.3 (C-7´), 117.9 (C-7), 114.8 (C-3´), 114.0 (C-5´), 111.3 (C-8), 101.9 (C-3), 60.6 (C-1´), 51.6 (NCH3)3 , 49.6 (C-1´´). Anal. calculated for: C, 54.04; H, 5.49; N, 6.63,. Found: C, 53.99; H, 5.63; N, 6.65. 1-(1-(4-hydroxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4c). Following the general procedure, compound 4c was obtained from 3c-1 as white solid. Yield: 98%. M. p: 153,5-158,4 °C decomposition. 27 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1 H NMR (400 MHz, DMSO-d6): 7.96 – 7.71 (m, 2H), 7.56 (d, J= 6.5 Hz, 1H), 7.31 – 7.00 (m, 4H), 6.71 (d, J = 6.5 Hz, 2H), 5.36 (s, 2H), 4.71 (s, 2H), 3.06 (s, 9H). (100 MHz, DMSO-d6): 13 C NMR 157.2 (C-5´), 136.2 (C-9), 133.6 (C-4), 129.1 (C-7´), 129.1 (C- 3´), 128.7 (C-2´), 127.9 (C-2), 122.3 (C-5), 120.7 (C-6), 119.2 (C-7), 115.7 (C-6´), 115.7 (C-4´), 111.3 (C-8), 101.7 (C-3), 60.6 (C-1´), 51.6 (C-1´´), 49.3 (-NCH3)3. Anal. calculated for: C, 54.04; H, 5.49; N, 6.63,. Found: C, 53.92; H, 5.61; N, 6.67. 1-(1-(4-methoxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4d). Following the general procedure, compound 4d was obtained from 3d-1 as white solid. Yield: 98%. M. p: 167,2-171,2 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.88 (d, J = 9.0 Hz, 2H), 7.57 (d, J= 7.7 Hz, 1H), 7.31 – 7.07 (m, 4H), 6.89 (d, J = 8.6 Hz, 2H), 5.43 (s, 2H), 4.73 (s, 2H), 3.70 (s, 3H), 3.07 (s, 9H). 28 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 13 C NMR (100 MHz, DMSO-d6): 159.0 (C-5´), 136.2 (C-9), 133.7 (C-2), 129.7 (C-4), 129.0 (C-7´), 129.0 (C-3´), 128.7 (C-2´), 122.4 (C-5), 120.7 (C-6), 119.3 (C-7), 114.3 (C6´), 114.3 (C-4´), 111.3 (C-8), 101.8 (C-3), 60.6 (C-1´´), 55.4 (C-8´), 51.6 (-NCH3)3, 49.2 (C-1´). Anal. calculated for: C, 55.05; H, 5.78; N, 6.42. Found: C, 54.92; H, 5.89; N, 6.44. 1-(1-(3-hydroxy-4-methoxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide( 4e). Following the general procedure, compound 4e was obtained from 4e-1 as white solid. Yield: 89%. M. p: 160,3-164,1 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.96 – 7.77 (m, 2H), 7.54 (d, J= 7.8 Hz, 1H), 7.24 – 7.13 (m, 2H), 6.86 (d, J = 8.3 Hz, 1H), 6.71 (dd, J = 8.2, 1.7 Hz, 1H), 6.63 (d, J = 1.8 Hz, 1H), 5.36 (s, 2H), 4.74 (s, 2H), 3.71 (s, 3H), 3.08 (s, 9H). 29 13 C NMR (100 MHz, DMSO- Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 d6): 147.4 (C-5´), 146.9 (C-4´), 136.3 (C-9), 133.8 (C-4), 130.3 (C-2), 128.6 (C-2´), 122.4 (C-7), 120.7 (C-6), 119.3 (C-7´), 118.5 (C-5), 114.8 (C-3´), 112.5 (C-6´), 111.3 (C-8), 101.7 (C-3), 60.6 (C-1´´), 56.0 (C-8´), 51.6 (-NCH3)3, 49.4 (C-1´). Anal. calculated for: C, 53.11; H, 5.57; N, 6.19. Found: C,53.00; H,5.63; N,6.25. 1-(1-(4-hydroxy-3-methoxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4f). Following the general procedure, compound 4f was obtained from 4f-1 as white solid. Yield: 85%. M. p: 158,5-162,9 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.87 (d, J = 5.5 Hz, 2H), 7.61 (d, J = 8.0 Hz, 1H), 7.18 (dt, J = 14.7, 7.0 Hz, 2H), 7.00 (s, 1H), 6.69 (q, J = 8.0 Hz, 2H), 5.36 (s, 2H), 4.73 (s, 2H), 3.73 (s, 3H), 3.07 (s, 9H). 13C NMR (400 MHz, DMSO-d6): 147.9 (C-4´), 146.4 (C-5´), 136.2 (C-9), 133.7 (C-4), 128.7 (C-2), 128.5 (C-2´), 122.3 (C-7), 120.7 (C-7´), 120.4 (C-6), 30 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 119.2 (C-5), 115.8 (C-6´), 112.3 (C-3´), 111.4 (C-8), 101.7 (C-3), 60.6 (C-1´´), 56.0 (C-8´), 51.5 (-NCH3)3, 49.6 (C-1´). Anal. calculated for: C, 53.11; H, 5.57; N, 6.19. Found: C,53.05; H,5.60; N,6.23. 1-(1-(3,4-dimethoxybenzyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4g). Following the general procedure, compound 4g was obtained from 3g-1 as white solid. Yield: 97%. M. p: 159.2-163.6 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.91 (d, J = 7.4 Hz, 1H), 7.81 (s, 1H), 7.30 (dd, J = 13.7, 4.3 Hz, 1H), 7.23 – 7.14 (m, 1H), 6.75 (d, J = 8.4 Hz, 1H), 6.62 (d, J = 8.2 Hz, 1H), 5.24 (s, 2H), 5.11 (s, 2H), 3.80 (s, 6H), 3.35 (s, 9H). 13C NMR (100 MHz, DMSO-d6): 149.2 (C- 3), 148.7 (C-5´), 136.4 (C-9), 133.5 (C-2´), 128.2 (C-4), 128.2 (C-2), 122.9 (C-7´), 121.5 31 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 (C-7), 119.4 (C-6), 111.4 (C-5), 110.5 (C-3), 110.4 (C-6´), 101.2 (C-8), 110.2 (C-3´), 62.0 (C-1´´), 56.2 (-NCH3), 56.0 (-N(CH3), 55.8 (-N(CH3), 52.5 (C-1´), 50.3 (C-8´), 50.3 (C-9´). Anal. calculated for: C, 54.08; H, 5.84; N, 6.01, Found: C,53.86; H,6.01; N,6,11. 1-(1-(benzo[d][1,3]dioxol-5-ylmethyl)-1H-indol-3-yl)-N,N,N-trimethylmethanaminium iodide (4h). Following the general procedure, compound 4h was obtained from 3h-1 as white solid. Yield: 98%. M. p: 173.6-180.2 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 8.53 (d, J = 6.5 Hz, 2H), 7.79 – 7.69 (m, 1H), 7.49 (t, J = 8.4 Hz, 2H), 7.28 – 7.18 (m, 3H), 5.71 (s, 2H), 4.61 (s, 2H), 4.20 (s, 2H), 3.03 (s, 9H). 13 C NMR (100 MHz, DMSO-d6): 145.1 (C-6´), 145.1 (C-5´), 135.9 (C-9), 133.7 (C-2´), 128.2 (C-4), 125.3 (C-2), 123.3 (C-3´), 122.6 (C-7), 121.3 (C-6), 121.3 (C-5), 119.1 (C-3), 119.1 (C-7´), 110.7 (C-4´), 110.7 (C-8), 102.6 (C-8´), 61.2 (C-1´´), 52.0 (-NCH3)3, 48.6 (C1´). Anal. calculated for: C, 53.34; H, 5.15; N, 6.22,. Found: C, 53.22; H, 5.21; N, 6.30. 32 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 N,N,N-trimethyl-4-((3-((trimethylammonio)methyl)-1H-indol-1yl)methyl)benzenaminium diiodide (4i). Following the general procedure, compound 4i was obtained from 3i-1 as white solid. Yield: 96%. M. p: 160,2-163,6 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 7.95 (dd, J = 21.8, 12.7 Hz, 4H), 7.56 (d, J = 7.7 Hz, 1H), 7.48 (d, J = 8.9 Hz, 2H), 7.29 – 7.11 (m, 2H), 5.65 (s, 2H), 4.78 (s, 2H), 3.59 (s, 9H), 3.11 (s, 9H). 13C NMR (100 MHz, DMSO-d6): 146.7 (C-5´), 140.1 (C-9), 136.1 (C-2´), 133.9 (C-4), 128.8 (C-7´), 128.8 (C-3´), 128.7 (C-2), 122.6 (C-5), 121.2 (C-6´), 121.2 (C4´), 121.0 (C-6), 119.5 (C-7), 111.2 (C-8), 102.3 (C-3), 60.5 (C-1´´), 56.8 (C-8´), 56.8 (C- 33 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 9´), 56.8 (C-10´), 51.6 (-NCH3)3, 48.6 (C-1´). Anal. calculated for: C, 56.90; H, 6.73; N, 9.05. Found: C, 56.85; H, 6.77; N, 9.11. 1-methyl-3-((3-((trimethylammonio)methyl)-1H-indol-1-yl)methyl)pyridin-1-ium iodide (4j). Following the general procedure, compound 4j was obtained from 3j-1 as white solid. Yield: 97%. M. p: 167,2-173,3 °C, decomposition. 1 H NMR (400 MHz, DMSO-d6): 9.15 (s, 1H), 8.95 (d, J = 5.9 Hz, 1H), 8.38 – 8.22 (m, 1H), 8.18 – 8.03 (m, 1H), 8.02 – 7.87 (m, 2H), 7.59 (t, J = 7.8 Hz, 1H), 7.33 – 7.09 (m, 2H), 5.79 (s, 2H), 4.79 (s, 2H), 4.37 (s, 3H), 3.11 (d, J = 13.2 Hz, 9H). MHz, DMSO-d6): 13 C NMR (100 144.7 (C-3´), 144.1 (C-5´), 143.3 (C-9), 137.7 (C-7´), 135.6 (C-2´), 133.5 (C-4), 128.5 (C-2), 127.6 (C-6´), 122.5 (C-5), 120.9 (C-6), 119.2 (C-7) , 110.7 (C-8), 34 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 102.5 (C-3), 59.9 (C-1´´), 51.3 (-NCH3)3, 48.2 (C-1´), 45.9 (C-8´). Anal. calculated for: C, 41.55; H, 4.59; N, 7.65. Found: C, 41.26; H, 4.60; N, 7.72. 35 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 1 H and 13C NMR spectrums 36 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 37 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 38 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 39 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 40 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 41 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 42 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 43 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 44 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 45 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 46 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 47 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 48 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 49 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 50 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 51 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 52 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 53 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 54 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 55 Electronic Supplementary Material (ESI) for Medicinal Chemistry Communications This journal is © The Royal Society of Chemistry 2013 56