Table of contents MS-Experiments... 3 Synthesis of intermediates and precursors... 4 Metabolic stability determination in vitro References...

Transcription

1 Supporting Information Trisubstituted Pyridinylimidazoles as Potent Inhibitors of the Clinically Resistant L858R/T790M/C797S EGFR Mutant: Targeting of Both Hydrophobic Regions and the Phosphate Binding Site Marcel Günther, 1, Jonas Lategahn, 2, Michael Juchum, 1 Eva Döring, 1 Marina Keul, 2 Julian Engel, 2 Hannah L. Tumbrink, 2 Daniel Rauh, 2 and Stefan Laufer 1,* 1 Institute of Pharmaceutical Sciences, Pharmaceutical and Medicinal Chemistry, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, Tübingen, Germany 2 Faculty of Chemistry and Chemical Biology, TU Dortmund University, Otto-Hahn-Straße 4a, Dortmund, Germany Table of contents MS-Experiments... 3 Synthesis of intermediates and precursors... 4 Metabolic stability determination in vitro References

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3 MS-Experiments Figure S1: 3

4 Synthesis of intermediates and precursors General Procedure A: A solution of sodium bis(trimethylsilyl)amide 2 M (2 eq) in THF was diluted with dry THF to a final concentration of 0.5 mol/l and cooled to 0 C. To the cooled solution 2-fluoro-4- methylpyridine (1 eq) was added via cannula under vigorous stirring. The resulting brownish solution was stirred for 45 minutes at 0 C after which the appropriate substituted benzoic acid ester (1.2 eq) was rapidly added via cannula under ice bath cooling. The mixture was allowed to warm up to room temperature over 45 minutes and subsequently poured into 2 M aq. HCl (15 eq) carefully. The resulting biphasic mixture was transferred into a separation funnel and the organic layer was separated, dried over Na 2 SO 4 and concentrated under reduced pressure to yield intermediates 32a-c as white solids. General Procedure B: Substituted phenylethan-1-ones obtained from general procedure A (1 eq.) were dissolved in glacial acetic acid under vigorous stirring. The reaction mixture was carefully cooled to 10 C. To the cooled solution sodium nitrite (3 eq.) dissolved in water (3.5 mol/l) was added dropwise over 30 minutes via cannula. The reaction mixture was allowed to slowly warm up to room temperature and stirred for additional 90 minutes. After complete consumption of the starting materials (TLC) the reaction was quenched by the addition of ice cold water. The resulting suspension was stirred for an additional hour under cooling and the product subsequently isolated by filtration. The analytically pure product was dried in vacuum overnight. General Procedure C: 4

5 Oxime (1 eq.) obtained from general procedure B was suspended in glacial acetic acid and vigorously sonicated to yield a clear solution. The mixture was carefully cooled to 0 C under rapid stirring. Zinc dust (1.5 eq.) was added in one portion. The reaction mixture was stirred until complete consumption of the starting material (HPLC monitoring). The resulting suspension was filtered and the solid was thoroughly washed with isopropyl alcohol. The filtrate was concentrated in vacuum to give an oily residue. The crude product was dissolved in ethyl acetate, cooled to -10 C and the pure amine was precipitated as the corresponding hydrochloric salts by passing HCl gas through this solution. The product was isolated by filtration and washed with cooled ethyl acetate. General Procedure D: α-aminoketone (1 eq.) obtained from general procedure C was dissolved in dry DMF. Potassium thiocyanate (2.5 eq.) was added to this solution and the mixture heated to reflux until HPLC indicated complete consumption of the starting material (usually 40 minutes). The reaction mixture was cooled to room temperature and quenched by the addition of ice/water. Vigorous stirring under cooling led to precipitation of the product that was isolated by filtration and thoroughly washed with water. The crude product was purified via silica flash chromatography. General Procedure E: The appropriate thione (1 eq.) obtained from general procedure D was dissolved in N- methylpyrrolidine. Under vigorous stirring, concentrated aq. HCl (4 eq.) was added at room temperature. The reaction mixture was refluxed for 30 minutes. Aniline (10 eq.) was added to the refluxing mixture dropwise. After complete addition, the reaction mixture is heated under reflux for further 90 minutes after which the reaction mixture is cooled to room temperature. The 5

6 resulting dark solution is added dropwise to a cooled sat. NH 4 Cl solution. The crude product precipitated and was isolated by filtration and washed with sat. NH 4 Cl solution. Purification was performed via recrystallization or silica flash chromatography General Procedure F: The appropriate alkyl halide (1.2 eq.), appropriate thione (1.0 eq.) and K 2 CO 3 (1.2eq.) were suspended in dry THF. The mixture was stirred vigorously and heated under reflux conditions until HPLC indicated complete consumption of the starting material. After cooling to room temperature, the solvent was evaporated. The residue was suspended in water, neutralized with NH4Cl and the crude product extracted three times with DCM. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed under reduced pressure. The crude material was purified via silica flash chromatography. General Procedure G: The appropriate phthalimide (1 eq.) obtained from general procedure F was dissolved in Ethanol. 80% Hydrazine hydrate (5 eq.) was added with stirring. The reaction mixture was stirred under reflux conditions until HPLC indicated complete consumption of the starting material. The solvents were removed under reduced pressure and the white residue was taken up in water. The product was extracted with EtOAc three times. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The crude product was purified via basic silica flash chromatography. General Procedure H: The Boc protected intermediate was dissolved in dry DCM. TFA was added under vigorous stirring at room temperature to reach a final acid concentration of 20%. The reaction mixture was stirred until TLC indicated complete consumption of the starting material (usually 60 minutes). All volatiles were removed via rotary evaporation. The residue was taken up in water, neutralized with NaHCO 3 and extracted three 6

7 times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The crude product was purified via basic silica flash chromatography. General Procedure I: The appropriate primary or secondary amine (1 eq.) obtained from general procedure G or H was suspended in dry THF. To the stirred solution was added the appropriate carboxylic acid (1 eq.), DIPEA (2 eq.) followed by TBTU (1.1 eq.). The reaction mixture was stirred until HPLC indicated complete consumption of the starting material. The reaction was quenched by the addition of sat. NH 4 Cl solution and extracted three times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The crude product was purified via silica flash chromatography General Procedure J: The appropriate commercial availlable non-racemic acid 38a-b (1 eq) was dissolved in dry THF (3 ml / mmol) and cooled to 0 C. BH 3 *THF(2.5 eq.) was added dropwise to the vigorously stirring solution at 0 C. The ice bath was removed after complete addition and the reaction mixture was stirred overnight. The reaction was quenched by the slow addition of Rochelle s salt (saturated aqueous solution). The aqueous phase was extracted thrice with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents evaporated to yield the title compound that was used in the next step without further purification. General Procedure K: Triphenylphosphine (1.5 eq.) dissolved in dry DCM (5 ml / mmol) was added dropwise to a cooled solution of the appropriate alcohol (1.0 eq.) and CBr 4 (1.5 eq.) in dry DCM (5 ml / mmol). After complete addition, the ice-bath was removed and the reaction mixture was stirred overnight. The resulting 7

8 suspension was filtered over celite and the filtrate was concentrated by rotary evaporation. The crude product was purified by silica flash chromatography. General Procedure L : Method A: aliphatic ketones The appropriate ketone (1 eq.) was dissolved in dry MeOH (2 ml / mmol). The solution was cooled under stirring to 0 C. Bromine (1 eq.) was added fast in one portion under vigorous stirring and the resulting dark solution was stirred overnight allowing to reach room temperature slowly. The reaction was quenched by the addition of water and sodium sulfite. The colorless solution was extracted three times with diethyl ether. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The crude product was used directly in the next step without further purification. Method B: aromatic ketones The appropriate ketone (1 eq.) was dissolved in dry DCM. Bromine (1 eq.) was added dropwise at room temperature under vigorous stirring. After complete addition, the mixture was stirred for additional 30 minutes at room temperature until decolorization. After addition of water and sodium sulfite the mixture was exracted three times with diethyl ether. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The crude product was used directly in the next step without further purification General Procedure M: S-Methyl isothiourea hemisulfate (1 eq.) was suspended in a mixture of THF/water (4:1). Under vigorous stirring NaHCO 3 (2.0 eq.) was added at room temperature. The reaction mixture was 8

9 heated under reflux for 15 minutes. The appropriate α-haloketone (1.0 eq.) dissolved in THF was added dropwise to the refluxing suspension. The mixture was heated for additional 30 minutes while the color changes from pale yellow to dark red. After cooling to room temperature the reaction mixture was filtered and the filtrate concentrated to dryness under reduced pressure. The dark residue was suspended in water and the ph adjusted to 8. The mixture was extracted five times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The product was purified via gradient silica flash chromatography General procedure N: The appropriate imidazole (1 eq.) obtained from general procedure M was dissolved in dry THF and cooled to -15 C using an ice-acetone bath. 60% Sodium hydride (1.1 eq) was added portion wise. The resulting suspension was allowed to warm up to room temperature. After stirring for 30 minutes at room temperature, the suspension was again cooled to -15 C and SEM-Cl (1.0 eq.) was added dropwise. After HPLC indicated complete consumption of the starting material (usually 30 minutes) the reaction was quenched by the addition of brine and EtOAc. The organic layer was washed with sat. NH 4 Cl solution followed by brine. The organic layer was dried over Na 2 SO 4, filtered and the volatiles removed under reduced pressure yielding a regioisomeric mixture of product. This crude mixture was dissolved in dry MeCN (0.3M). A catalytic amount (5 mol%)of SEM-Cl was added to the solution, which was stirred at 70 C overnight. After cooling to room temperature, the volatiles were removed by rotary evaporation. The residue was taken up on celite and subjected to purification via silica flash chromatography. General Procedure O: SEM protected imidazole (1.0 eq.) obtained from general Procedure N was dissolved in dry MeCN (0.1M). The resulting solution or suspension was cooled to -30 C. NBS (1.05 eq) 9

10 dissolved in MeCN (0.5 M) was added dropwise to the vigorous stirring cooled solution. The reaction mixture was stirred at -30 C until HPLC indicated complete consumption of the starting material (usually 15 minutes). The reaction was quenched at -30 C by addition of saturated sodium sulfite solution and brine. The mixture was extracted three times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvent removed by rotary evaporation. The product was purified via gradient silica flash chromatography. General Procedure P: Method A: 5-bromoimidazole (1 eq.) obtained from general procedure O was suspended in degassed 1-4- dioxane (3 ml / mmol). A previously degassed aqueous solution of K 3 PO 4 (1 M, 4 eq.) was added under vigorous stirring. The reaction mixture was carefully degassed under vacuum and backfilled with argon. Pd(OAc) 2 (2.5 mol %) and XPhos (5 mol %) were added under an argon flow and the mixture was heated to reflux. The appropriate pyridine boronic acid ester (2.2 eq.) was added to the refluxing reaction mixture in small protions over 30 minutes. After complete addition, the biphasic dark solution was heated under reflux for additional 60 minutes. The resulting suspension was cooled to room temperature, quenched by the addition of a saturated NH 4 Cl solution and extracted three times with DCM. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. Method B: 5-bromoimidazole (1 eq.) obtained from general procedure O and the appropriate pyridine boronic acid ester (1.3 eq.) and K 3 PO 4 (3 eq.) were suspended in degassed 1-4-dioxane (3 ml / 10

11 mmol). Pd 2 (dba) 3 (5 mol%) and (t-bu) 3 P*HBF 4 (5 mol%) were added and the mixture was stirred under argon for 5 minutes. Previously degassed water (1.5 ml / mmol) was added under vigorous stirring. The reaction mixture was carefully degassed under vacuum and backfilled with argon. After complete addition, the biphasic solution was stirred at 40 C until total consumption of the starting material (usually overnight). The resulting suspension was cooled to room temperature, quenched by the addition of a saturated NH 4 Cl solution and extracted three times with DCM. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. General Procedure Q: The appropriate acetamide (1.0 eq.) obtained from general procedure N was dissolved in methanol (20 ml / mmol). A solution of 5 M aqueous NaOH (25 eq.) was added to the vigorously stirred solution via canula at room temperature. The reaction was stirred at room temperature until TLC indicated complete consumption of the starting material (usually 5-7 hours). The mixture was neutralized with saturated aqueous NH 4 Cl. Methanol was removed via rotary evaporation and the residue suspended in water. The aqueous phase was extracted three times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. General Procedure R: The appropriate dimethylpyrrole protected 2-Aminopyridine (1 eq.) obtained from general procedure P was dissolved in a mixture of ethanol/water (2:1; 5 ml / mmol). To the vigorously stirred solution, hydroxylamine hydrochloride (6 eq.) followed by DIPEA (6 eq.) was added at 11

12 room temperature. The reaction vessel was sealed with a rubber septum and carefully heated to 65 C until TLC indicated complete conversion of the starting material (usually 48 hours). The reaction mixture was cooled to room temperature and the solvents removed under reduced pressure. The resulting residue was taken up in saturated aqueous NaHCO 3 solution and extracted three times with ethyl acetate. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. General Procedure S: The appropriate Boc protected 2-Aminopyridine (1 eq.) obtained from general Procedure P was dissolved in 5% TFA in dry DCM (2 ml / mmol). The resulting solution was stirred at room temperature until total consumption of the starting material (usually overnight). The reaction was quenched by the addition of a saturated aqueous NaHCO 3 solution and extracted three times with ethyl acetate. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. General Procedure T: The appropriate 2-Aminopyridine (1.0 eq.) obtained from general procedure Q, R or S was dissolved in a mixture of dry 1,4-dioxane and tert butyl alcohol (4:1) to give a final concentration of 0.1 mol/l based on the starting material. The appropriate substituted bromo(hetero)arene (1.2 eq.) was added to the reaction mixture under vigorous stirring followed by cesium carbonate (5.0 eq.). The resulting suspension was carefully degassed under vacuum and backfilling with argon. Brettphos Pd G1 (2.5 mol%) was added under a flow of argon to the reaction mixture. The 12

13 suspension was heated under reflux until TLC indicated complete consumption of the starting material (usually minutes). After cooling to room temperature, the solvents were removed by rotary evaporation. The residue was either taken up on celite and purified by gradient silica flash chromatography or directly subjected to deprotection. General Procedure U: The appropriate SEM-protected precursor (1.0 eq.) was dissolved or suspended in 20% TFA in dry DCM (3 ml / mmol). The mixture was stirred overnight or until HPLC indicated complete consumption of the starting material. All volatiles were removed via rotary evaporation and the residue was dissolved in EtOAc. This solution was poured into a saturated aqueous NaHCO 3 solution and extracted three times with EtOAc. The combined organic layers were dried over Na 2 SO 4, filtered and the solvents removed by rotary evaporation. The product was purified via gradient silica flash chromatography. 2-(2-fluoropyridin-4-yl)-1-phenylethan-1-one (36b): The title compound was prepared from ethyl benzoate according to general procedure A and obtained as a white solid in 65 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ 8.19 (d, J = 5.1, 0.7 Hz, 1H), (m, 2H), (m, 3H), (m, 1H), 7.12 (s, 1H), 4.60 (s, 2H); 13 C NMR (50 MHz, DMSO-d 6 ) δ 196.3, (d, J=234 Hz), 151.2, 151.1, 147.5, 147.2, 136.5, 133.9, 129.1, 128.6, 124.2, 124.1, 111.5, 110.8, 44.1, (4-fluoro-3-(trifluoromethyl)phenyl)-2-(2-fluoropyridin-4-yl)ethan-1-one (36c): 13

14 The title compound was prepared from ethyl (4-fluoro-3-(trifluoromethyl)benzoate according to general procedure A and obtained as a white solid in 79 % yield. 1 H NMR (200 MHz, Chloroform-d) δ (m, 3H), (m, 1H), (m, 1H), 6.88 (s, 1H), 4.39 (s, 2H). LRMS (FAB-MS): [M+H] + 2-(2-fluoropyridin-4-yl)-2-(hydroxyimino)-1-phenylethan-1-one (37b) The title compound was prepared from 36b according to general procedure B and obtained as a white solid in 97% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 8.29 (d, J = 5.3, 1H), (m, 2H), (m, 1H), (m, 2H), (m, 1H), (m, 1H); 13 C NMR (50 MHz, DMSO-d 6 ) δ 194.0, (d, J =235 Hz), 152.6, 152.5, 149.3, 149.0, 144.9, 144.7, 135.4, 134.3, 129.8, 129.3, 118.6, 118.5, 106.1, (4-fluoro-3-(trifluoromethyl)phenyl)-2-(2-fluoropyridin-4-yl)-2-(hydroxyimino)ethan-1- one(37c) The title compound was prepared from 36c according to general procedure B and obtained as a yellow solid in 86% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), (m, 3H), (m, 1H), (m, 1H), 7.26 (s, 1H). 1-(2-fluoropyridin-4-yl)-2-oxo-2-phenylethan-1-aminium chloride (38b): The title compound was prepared from 37b according to general procedure C and obtained as a white solid in 85 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ 8.97 (bs, 3H), (m, 1H), (m, 2H), (m, 1H), (m, 4H), 6.48 (bs, 1H). 2-(4-fluoro-3-(trifluoromethyl)phenyl)-1-(2-fluoropyridin-4-yl)-2-oxoethan-1-aminium chloride (38c): 14

15 The title compound was prepared from 37c according to general procedure C and obtained as a white solid in 78 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ 9.06 (s, 3H), (m, 1H), (m, 2H), (m, 1H), (m, 1H), (m, 1H), 5.08 (s, 7H). LRMS (FAB-MS): [M-Cl - ] + 4-(2-fluoropyridin-4-yl)-5-phenyl-1,3-dihydro-2H-imidazole-2-thione (39b): The title compound was prepared from 38b according to general procedure D and obtained as a yellow solid after gradient silica flash chromatography (DCM DMC/MeOH 15%) in 84 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 2H), 8.08 (d, J = 5.3 Hz, 1H), (m, 5H), (m, 2H). 13 C NMR (50 MHz, DMSO) δ (d, J = 234 Hz), 162.9, (d, J = 16 Hz), (d, J = 9 Hz), 129.7, 129.5, 129.4, 129.1, 128.2, (d, J = 4 Hz), (d, J = 4 Hz), (d, J = 40 Hz). LRMS (FAB-MS): [M+H] + 4-(2-fluoropyridin-4-yl)-5-(-(4-fluoro-3-(trifluoromethyl))-1,3-dihydro-2H-imidazole-2-thione (39c): The title compound was prepared from 38c according to general procedure D and obtained as a brown solid after gradient silica flash chromatography (DCM DMC/MeOH 10%) in 73 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 2H), 8.13 (d, J = 5.3 Hz, 1H), (m, 1H), (m, 1H), (m, 1H), (m, 2H). LRMS (FAB-MS): [M+H] + 4-phenyl-5-(2-(phenylamino)pyridin-4-yl)-1,3-dihydro-2H-imidazole-2-thione (40b): The title compound was prepared from 39b according to general procedure E and obtained as a yellow solid after recrystallization from DMF/H 2 O in 63 % yield. 15

16 thione (40c): The title compound was prepared from 39c according to general procedure E and obtained as a yellow solid after silica flash chromatography (Hexane/EtOAc 2:1) in 66 % yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 2H), 9.09 (s, 1H), 8.11 (d, J = 5.4 Hz, 1H), (m, 1H), 7.72 (s, 1H), 7.51 (d, J = 8.2 Hz, 3H), 7.21 (t, J = 7.7 Hz, 2H), (m, 3H). yl)thio)ethyl)isoindoline-1,3-dione (41a): The title compound was synthesized from 40a and 2-(3-bromoethyl)isoindoline-1,3-dione according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (DCM DCM/iPrOH 2.5%) in 62% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), (m, 1H), (m, 1H), (m, 6H), (m, 6H), (m, 2H), (m, 1H), 3.97 (t, J = 6.0 Hz, 2H), 3.47 (t, J = 6.1 Hz, 2H). LRMS (FAB-MS): [M+H] + 4-(2-(phenylamino)pyridin-4-yl)-5-(4-fluoro-3-(trifluoromethyl))-1,3-dihydro-2H-imidazole-2-2-(2-((4-(4-fluorophenyl)-5-(2-(phenylamino)pyridin-4-yl)-1H-imidazol-2-2-(3-((4-(4-fluorophenyl)-5-(2-(phenylamino)pyridin-4-yl)-1H-imidazol-2- yl)thio)propyl)isoindoline-1,3-dione (41b): The title compound was synthesized from 40a and 2-(2-bromopropyll)isoindoline-1,3-dione according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (DCM DCM/iPrOH 2.5%) in 57% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), (m, 1H), (m, 1H), (m, 4H), (m, 4H), 16

17 (m, 4H), 7.01 (s, 1H), (m, 1H), (m, 1H), (m, 2H), 3.15 (t, J = 7.2 Hz, 2H), 2.02 (p, J = 6.8 Hz, 2H). LRMS (FAB-MS): [M+H] + yl)thio)ethyl)isoindoline-1,3-dione (41c): The title compound was synthesized from 40c and 2-(3-bromoethyl)isoindoline-1,3-dione according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (DCM DCM/iPrOH 2%) in 80% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 9.00 (s, 1H), (m, 1H), (m, 8H), 7.21 (t, J = 7.7 Hz, 2H), (m, 3H), (m, 2H), (m, 2H). LRMS (FAB-MS): [M+H] + dione (41d): The title compound was synthesized from 40b and 2-(3-bromoethyl)isoindoline-1,3-dione according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (DCM DCM/iPrOH 2.5%) in 61% yield. 1 H NMR (200 MHz, Chloroform-d) δ (bs, 1H), 7.98 (d, J = 5.6 Hz, 1H), 7.81 (dd, J = 5.5, 3.0 Hz, 2H), 7.66 (dd, J = 5.5, 3.1 Hz, 2H), (m, 5H), (m, 7H), 3.98 (t, J = 5.7 Hz, 2H), (bs, 1H) 3.48 (q, J = 7.0 Hz, 1H), 3.26 (t, J = 5.7 Hz, 2H). LRMS (FAB-MS): [M+H] + 2-(2-((5-(2-(phenylamino)pyridin-4-yl)-4-(4-fluoro-3-(trifluoromethyl))-1H-imidazol-2-2-(2-((4-phenyl-5-(2-(phenylamino)pyridin-4-yl)-1H-imidazol-2-yl)thio)ethyl)isoindoline-1,3- tert-butyl-(r)-2-(((4-(4-fluorophenyl)-5-(2-(phenylamino)pyridin-4-yl)-1h-imidazol-2- yl)thio)methyl)pyrrolidine-1-carboxylate (46a): 17

18 The title compound was synthesized from 40a and 45a according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (Hexane Hexane/EtOAc 5:1) in 80% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 8.98 (s, 1H), (m, 1H), (m, 4H), (m, 4H), (m, 1H), 6.85 (t, J = 7.3 Hz, 1H), (m, 1H), 3.95 (s, 1H), (m, 4H), (m, 4H), 1.24 (s, 9H). LRMS (FAB-MS): [M+H] + tert-butyl (S)-2-(((4-(4-fluorophenyl)-5-(2-(phenylamino)pyridin-4-yl)-1H-imidazol-2-yl)thio) methyl)pyrrolidine-1-carboxylate (46b): The title compound was synthesized from 40a and 45b according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (Hexane Hexane/EtOAc 5:1) in 76% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 9.04 (s, 1H), (m, 1H), (m, 4H), (m, 5H), (m, 2H), (m, 1H), (m, 5H), (m, 3H), 1.28 (s, 9H). LRMS (FAB-MS): [M+H] + tert-butyl 3-((4-(4-fluorophenyl)-5-(2-(phenylamino)pyridin-4-yl)-1H-imidazol-2-yl)thio)pyrrol idine-1-carboxylate (50): The title compound was synthesized from 40a and 49 according to general procedure F and obtained as an off-white solid after gradient silica flash chromatography (DCM DCM/iPrOH 5%) in 84% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), (m, 1H), (m, 1H), (m, 4H), (m, 5H), (m, 2H), (m, 1H), (m, 4H), (m, 1H), (m, 1H), 1.37 (s, 9H). 13 C NMR (50 MHz, 18

19 DMSO-d 6 ) δ 162.6, (d, J = 246 Hz), 156.6, 153.7, 147.6, 143.1, 142.1, 139.1, 135.8, (d, J = 8.1 Hz), 130.4, 128.8, (d, J = 3.3 Hz), 118.6, 118.4, (d, J = 22 Hz, 112.5, 107.6, 78.9, 62.4, 52.1, 36.1, 31.1, LRMS (FAB-MS): [M+H] + 4-(2-((2-aminoethyl)thio)-4-(phenyl)-1H-imidazol-5-yl)-N-phenylpyridin-2-amine (42f): The title compound was synthesized from 41d according to general procedure G and obtained as a white solid after silica flash chromatography (EtOAc/MeOH/NH 3 90:8:2) in 65% yield. 1 H NMR (200 MHz, MeOH-d 4 ) δ 7.92 (d, J = 5.5 Hz, 1H), (m, 6H), (m, 5H), 6.85 (d, J = 6.0 Hz, 2H), 6.74 (d, J = 5.6 Hz, 1H), 3.06 (t, J = 6.1 Hz, 2H), 2.85 (t, J = 6.1 Hz, 2H). 4-(2-((2-aminoethyl)thio)-4-(4-fluoro-3-(trifluoromethyl)phenyl)-1H-imidazol-5-yl)-N-phenyl pyridin-2-amine (42e): The title compound was synthesized from 41c according to general procedure G and obtained as a white solid after silica flash chromatography (EtOac/MeOH/NH 3 90:8:2) in 58% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ 8.99 (bs, 1H), (m, 1H), (m, 2H), (m, 3H), (m, 2H), (m, 3H), 5.84 (bs, 3H), (m, 2H), (m, 2H). (R)-4-(4-(4-fluorophenyl)-2-((pyrrolidin-2-ylmethyl)thio)-1H-imidazol-5-yl)-N-phenylpyridin-2- amine (47a): The title compound was synthesized from 46a according to general procedure H and obtained as a white solid after basic silica flash chromatography (Hexane/MeOH/NH 3 90:8:2) in 88% yield. 19

20 1 H NMR (200 MHz, Chloroform-d) δ 9.47 (s, 2H), 7.96 (s, 1H), (m, 12H), (m, 3H), (m, 5H). 13 C NMR (50 MHz, Chloroform-d) δ (d, J = 248 Hz), 156.6, 147.9, 142.5, 141.4, 140.4, 134.0, 131.9, (d, J = 8.1 Hz), 128.9, (d, J = 3.3 Hz), 122.2, 119.9, (d, J = 22 Hz), 112.2, 105.8, 59.2, 45.0, 37.3, 30.0, LRMS (FAB- MS): [M+H] + (S)-4-(4-(4-fluorophenyl)-2-((pyrrolidin-2-ylmethyl)thio)-1H-imidazol-5-yl)-N-phenylpyridin-2- amine (47b): The title compound was synthesized from 46b according to general procedure H and obtained as a white solid after basic silica flash chromatography (EtOAc/MeOH/NH 3 90:8:2) in 86% yield. 1 H NMR (200 MHz, Chloroform-d) δ 9.48 (s, 2H), 7.97 (s, 1H), (m, 12H), (m, 3H), (m, 5H). 13 C NMR (50 MHz, Chloroform-d) δ (d, J = 248 Hz), 156.8, 148.1, 142.7, 141.6, 140.6, 134.2, 132.1, (d, J = 8.1 Hz), 129.1, (d, J = 3.3 Hz), 122.4, 120.1, (d, J = 22 Hz), 112.4, 106.0, 59.4, 45.2, 37.5, 30.2, LRMS (FAB- MS): [M+H] + 4-(4-(4-fluorophenyl)-2-(pyrrolidin-3-ylthio)-1H-imidazol-5-yl)-N-phenylpyridin-2-amine (51): The title compound was synthesized from 50 according to general procedure H and obtained as a white solid after basic silica flash chromatography (EtOAc/MeOH/NH 3 90:8:2) in 81% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), (m, 3H), (m, 1H), (m, 3H), (m, 3H), (m, 3H), (m, 1H), (m, 1H), (m, 1H), (m, 2H), (m, 3H). 13 C NMR (50 MHz, DMSO-d 6 ) δ (d, J = 246 Hz), 159.3, 158.7, 155.9, 146.6, 141.3, 139.1, (d, J = 8.6 Hz), 129.0, 20

21 121.5, 120.4, 119.2, (d, J = 21.6 Hz), 114.5, 112.6, 108.5, 107.6, 50.6, 44.6, 43.3, LRMS (FAB-MS): [M+H] + tert-butyl (S)-2-(bromomethyl)pyrrolidine-1-carboxylate (45b): The title compound was prepared from 44b according to general Procedure K and obtained as a white solid after gradient silica flash chromatography (hexane hexane/etoac 6:1) in 71 % yield. 1 H NMR (200 MHz, Chloroform-d) δ (m, 1H), (m, 1H), (m, 3H), (m, 2H), (m, 2H), 1.40 (s, 9H). tert-butyl 3-bromopyrrolidine-1-carboxylate (49): The title compound was prepared from 48 according to general Procedure K and obtained as a yellow solid after gradient silica flash chromatography (hexane hexane/etoac 5:1) in 62 % yield. 1 H NMR (200 MHz, Chloroform-d) δ (m, 1H), (m, 4H), (m, 2H), 1.40 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) major rotamere: δ 154.0, 79.4, 55.6, 47.1, 43.7, 35.6, bromo-1-phenylethan-1-one (52b): The title compound was prepared from acetophenone according to general Procedure L (method B) in 98% yield and was used in the next step without further purification. 1 H NMR (200 MHz, DMSO-d 6 ) δ (m, 2H), (m, 3H), 4.93 (s, 2H). 13 C NMR (50 MHz, DMSOd 6 ) δ 191.6, 133.9, 133.8, 128.8, 128.7, LRMS: not detected 2-bromo-1-(naphthalen-2-yl)ethan-1-one (52c): 21

22 The title compound was prepared from 1-(naphthalen-1-yl)ethan-1-one according to general Procedure L (method B) in 95% yield and was used in the next step without further purification. 1 H NMR (200 MHz, DMSO-d 6 ) δ (m, 1H), (m, 4H), (m, 2H), 5.07 (d, J = 0.9 Hz, 2H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 192.0, 135.6, 132.4, 131.7, 131.4, 130.0, 129.4, 128.9, 128.1, 127.5, 124.3, LRMS: not detected 2-bromo-1-(thiophen-2-yl)ethan-1-one (82d): The title compound was prepared from 1-(thiophen-2-yl)ethan-1-one according to general Procedure L (method B) in 82% yield and was used in the next step without further purification. 1 H NMR (200 MHz, DMSO-d 6 ) δ (m, 1H), (m, 1H), (m, 1H), 4.34 (s, 2H). LRMS (ESI-MS): 204.9/206.9 [M+H] + 2-bromo-1-cyclobutylethan-1-one (52h): The title compound was prepared from 1-cyclobutylethan-1-one according to general Procedure L (method A) in 64% yield and was used in the next step without further purification. 1 H NMR (200 MHz, Chloroform-d) δ (m, 2H), (m, 1H), (m, 6H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 202.0, 51.0, 43.3, 25.4, LRMS: not detected. 2-bromo-1-cyclopentylethan-1-one (52i): The title compound was prepared from 1-cyclopentylethan-1-one according to general Procedure L (method A) in 89% yield and was used in the next step without further purification. 1 H NMR (200 MHz, Chloroform-d) δ 3.97 (m, 2H), (m, 1H), (m, 8H). 13 C NMR (50 MHz, Chloroform-d) δ 204.4, 48.7, 34.3, 29.6, LRMS: not detected 22

23 1-bromo-3,3-dimethylbutan-2-one (52j): The title compound was prepared from 1-cyclopentylethan-1-one according to general Procedure L (method A) in 93% yield and was used in the next step without further purification. 1 H NMR (200 MHz, Chloroform-d) δ 4.16 (s, 2H), 1.21 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 206.1, 44.3, 31.8, LRMS: not detected. 2-(methylthio)-4-phenyl-1H-imidazole (43b): The title compound was prepared from 52b according to general Procedure M and obtained as a white solid after gradient silica flash chromatography (hexane hexane/etoac 1:1) in 43% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (m, 1H), (m, 2H), (m, 3H), (m, 3H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 143.4, 134.3, 129.3, 129.3, 127.3, 125.7, 117.1, LRMS (FAB-MS): [M+H] + 2-(methylthio)-4-(naphthalen-2-yl)-1H-imidazole (53c): The title compound was prepared from 52c according to general Procedure M and obtained as a white solid after gradient silica flash chromatography (hexane hexane/etoac 2:1) in 61% yield. The crude product was used in the next step without further purification. 1 H NMR (200 MHz, Chloroform-d) δ (s, 1H), (m, 3H), (m, 5H), 2.60 (s, 3H). LRMS (ESI-MS): [M+H] + 2-(methylthio)-4-(thiophen-2-yl)-1H-imidazole (53d): The title compound was prepared from 52d according to general Procedure M and obtained as a pale yellow solid after gradient silica flash chromatography (hexane hexane/etoac 2:1) in 31% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 7.44 (s, 1H), (m, 1H),

24 7.19 (m, 1H), (m, 1H), 2.54 (s, 3H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 141.5, 131.2, 127.9, 123.5, 121.6, LRMS (ESI-MS): [M+H] + 2-(methylthio)-1H-imidazole (53e): The title compound was prepared from commercially available 1,3-dihydro-2H-imidazole-2-thione by methylation as it is described in the literature. 1 4-methyl-2-(methylthio)-1H-imidazole (53f): The title compound was prepared from chloroacetone according to general Procedure M and obtained as a brown semi solid in 44% yield. The crude product was used in the next step without further purification. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 7.14 (t, J = 1.1 Hz, 1H), 2.68 (s, 3H), 2.18 (s, 3H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 140.4, 131.4, 118.2, 16.3, LRMS (ESI-MS): [M+H] + 4-cyclopropyl-2-(methylthio)-1H-imidazole (53g): The title compound was prepared from 52g according to general Procedure M and obtained as a pale yellow solid after gradient silica flash chromatography (hexane hexane/etoac 2:3) in 43% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (s, 1H), 6.72 (s, 1H), 2.43 (s, 3H), (m, 1H), (m, 2H), (m, 2H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 139.1, 16.2, 7.1. LRMS (ESI-MS): [M+H] + 4-cyclobutyl-2-(methylthio)-1H-imidazole (53h): The title compound was prepared from 52h according to general Procedure M and obtained as a grey solid after gradient silica flash chromatography (hexane hexane/etoac 2:1) in 15% yield. 24

25 1 H NMR (200 MHz, Chloroform-d) δ (s, 1H), 6.84 (s, 1H), (m, 1H), (m, 3H), (m, 6H). 13 C NMR (50 MHz, Chloroform-d) δ 142.3, 140.3, 119.0, 32.5, 29.1, 18.4, LRMS (ESI-MS): [M-H] - 4-cyclopentyl-2-(methylthio)-1H-imidazole (53i): The title compound was prepared from 52i according to general Procedure M and obtained as an off-white solid after gradient silica flash chromatography (hexane hexane/etoac 1:1) in 40% yield. 1 H NMR (200 MHz, Chloroform-d) δ (s, 1H), 6.78 (s, 1H), (m, 1H), 2.50 (d, J = 1.1 Hz, 3H), (m, 2H), (m, 6H). 13 C NMR (50 MHz, Chloroform-d) δ 141.9, 140.0, 119.1, 37.4, 32.7, 24.9, LRMS (ESI-MS): [M+H] + 4-(tert-butyl)-2-(methylthio)-1H-imidazole (53j): The title compound was prepared from 52j according to general Procedure M and obtained as an off-white solid after gradient silica flash chromatography (hexane hexane/etoac 2:1) in 21% yield. 1 H NMR (200 MHz, Chloroform-d) δ (s, 1H), 6.77 (s, 1H), 2.49 (s, 3H), 1.29 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 146.9, 140.1, 118.5, 31.0, 30.0, LRMS (ESI-MS): [M-H] - 2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54b): The title compound was prepared from 53b according to general Procedure N and obtained as a colorless oil after gradient silica flash chromatography (hexane hexane/etoac 1:1) in 86% yield. 1 H NMR (200 MHz, Chloroform-d) δ 7.04 (d, J = 5.9 Hz, 2H), 5.24 (s, 2H), (m, 2H), 2.59 (s, 2H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 143.5, 129.4, 120.9, 74.7, 66.2, 17.6, 16.1, LRMS (ESI-MS): [M+H] + 25

26 2-(methylthio)-4-(naphthalen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54c): The title compound was prepared from 53c according to general Procedure N and obtained as a waxy solid after gradient silica flash chromatography (hexane hexane/etoac 4:1) in 76% yield. 1 H NMR (200 MHz, Chloroform-d) δ 8.38 (s, 1H), (m, 4H), (m, 3H), 5.26 (s, 2H), (m, 2H), 2.72 (s, 3H), (m, 2H), 0.04 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 144.0, 142.1, 133.7, 132.6, 131.1, 128.1, 128.0, 127.6, 126.0, 125.3, 123.4, 122.9, 117.1, 75.0, 66.3, 17.7, 16.8, LRMS (ESI-MS): [M+H] + 2-(methylthio)-4-(thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54d): The title compound was prepared from 53d according to general Procedure N and obtained as a brownishoil after gradient silica flash chromatography (hexane hexane/etoac 3:1) in 68% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ 7.69 (s, 1H), (m, 1H), (m, 1H), (m, 1H), 5.26 (s, 2H), 3.52 (t, J = 8.0 Hz, 2H), 2.56 (s, 3H), 0.85 (t, J = 8.0 Hz, 2H), (m, 10H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 143.3, 137.9, 136.4, 128.0, 123.9, 121.9, 117.8, 74.9, 65.9, 17.5, 16.3, LRMS (ESI-MS): [M+H] + 2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54e): The title compound was prepared from 53e according to general Procedure N and obtained as a colorless waxy solid after gradient silica flash chromatography (hexane hexane/etoac 4:1) in 87% yield. 1 H NMR (200 MHz, Chloroform-d) δ (m, 2H), 5.24 (d, J = 0.7 Hz, 2H), (m, 2H), 2.59 (s, 3H), (m, 2H), (s, 9H). LRMS (ESI-MS): [M+H] + 26

27 4-methyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54f): The title compound was prepared from 53f according to general Procedure N and obtained as a yellow gum after gradient silica flash chromatography (hexane hexane/etoac 4:1) in 50% yield. 1 H NMR (200 MHz, Chloroform-d) δ 6.75 (s, 1H), 5.18 (s, 2H), 3.46 (t, J = 8.1 Hz, 2H), 2.51 (s, 3H), 2.19 (s, 3H), 0.85 (t, J = 8.1 Hz, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 142.6, 129.5, 127.1, 72.4, 65.8, 17.6, 16.5, 9.6, LRMS (ESI-MS): [M+H] + 4-cyclopropyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54g): The title compound was prepared from 53g according to general Procedure N and obtained as a yellow gum after gradient silica flash chromatography (hexane hexane/etoac 3:1) in 58% yield. 1 H NMR (200 MHz, Chloroform-d) δ 6.70 (s, 1H), 5.16 (s, 2H), (m, 2H), 2.52 (s, 3H), (m, 1H), (m, 6H), (s, 9H). 4-cyclobutyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54h): The title compound was prepared from 53h according to general Procedure N and obtained as a waxy solid after gradient silica flash chromatography (hexane hexane/etoac 5:1) in 83% yield. 1 H NMR (200 MHz, Chloroform-d) δ 6.83 (s, 1H), 5.11 (s, 2H), (m, 3H), 2.53 (s, 3H), (m, 6H), (m, 2H), (s, 9H). 4-cyclopentyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54i): The title compound was prepared from 53i according to general Procedure N and obtained as a colorless oil after gradient silica flash chromatography (hexane hexane/etoac 3:1) in 69% yield. 1 H NMR (200 MHz, Chloroform-d) δ 6.75 (s, 1H), 5.20 (s, 2H), 3.46 (t, J = 8.4 Hz, 2H), 27

28 (m, 1H), 2.50 (s, 3H), (m, 2H), (m, 6H), 0.85 (t, J = 8.1 Hz, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 142.8, 138.7, 124.8, 72.5, 65.7, 35.1, 32.6, 24.9, 17.7, 16.4, LRMS (ESI-MS): [M+H] + 4-(tert-butyl)-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (54j): The title compound was prepared from 53j according to general Procedure N and obtained as a colorless oil after gradient silica flash chromatography (hexane hexane/etoac 5:1) in 93% yield. 1 H NMR (200 MHz, Chloroform-d) δ 6.75 (s, 1H), 5.24 (s, 2H), (m, 2H), 2.53 (s, 3H), 1.26 (s, 9H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 152.8, 141.5, 114.7, 74.8, 66.0, 31.7, 29.8, 17.7, 17.6, LRMS (ESI-MS): [M+H] + 5-bromo-2-(methylthio)-4-(naphthalen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55c): The title compound was prepared from 54c according to general Procedure O and obtained as a colorless oil after gradient silica flash chromatography (hexane hexane/etoac 9:1) in 75% yield. 1 H NMR (200 MHz, Chloroform-d) δ 8.50 (s, 1H), 8.19 (dd, J = 8.7, 1.8 Hz, 1H), (m, 3H), (m, 2H), 5.34 (d, J = 2.0 Hz, 2H), 3.66 (t, J = 8.2 Hz, 2H), 2.70 (s, 3H), 0.98 (t, J = 8.3 Hz, 2H), 0.03 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 177.9, 145.1, 139.1, 133.3, 132.6, 130.2, 128.2, 127.7, 127.5, 126.0, 125.8, 125.5, 124.8, 100.7, 73.7, 66.5, 29.4, 17.8, 16.3, LRMS (ESI-MS): 449.1/501.1 [M+H] + 5-bromo-2-(methylthio)-4-(thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55d): 28

29 The title compound was prepared from 54d according to general Procedure O and obtained as a brown oil after gradient silica flash chromatography (hexane hexane/etoac 8:1) in 79% yield. 1 H NMR (200 MHz, DMSO-d 6 ) δ (m, 2H), 7.10 (dd, J = 5.1, 3.6 Hz, 1H), 5.29 (s, 2H), 3.57 (t, J = 7.7 Hz, 2H), 2.59 (s, 3H), 0.86 (t, J = 7.7 Hz, 2H), (s, 10H). 13 C NMR (50 MHz, DMSO-d 6 ) δ 145.2, 135.8, 134.8, 127.9, 125.3, 123.6, 100.2, 74.0, 66.3, 17.6, 15.9, LRMS (ESI-MS): 405.0/407.0 [M+H] + 4-bromo-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55e): The title compound was prepared from 54e according to general Procedure O and obtained as a yellow oil after gradient silica flash chromatography (hexane hexane/etoac 2:1) in 21% yield. 1 H NMR (200 MHz, Chloroform-d) δ 7.01 (s, 1H), 5.20 (s, 2H), (m, 2H), 2.61 (s, 3H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 145.0, 129.8, 119.9, 73.3, 66.5, 17.7, 16.0, LRMS: not detected. 5-bromo-4-methyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55f): The title compound was prepared from 54f according to general Procedure O and obtained as a yellow gum after gradient silica flash chromatography (hexane hexane/etoac 6:1) in 90% yield. 1 H NMR (200 MHz, Chloroform-d) δ 5.27 (s, 2H), (m, 2H), 2.59 (s, 3H), 2.21 (s, 3H), (m, 2H), 0.01 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 143.6, 137.2, 101.3, 73.7, 66.4, 17.7, 16.6, 12.8, LRMS (ESI-MS): 337.0/339.0 [M+H] + 5-bromo-4-cyclopropyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55g): The title compound was prepared from 54g according to general Procedure O and obtained as colorless sticky oil after gradient silica flash chromatography (hexane hexane/etoac 8:1) in 29

30 72% yield. 1 H NMR (200 MHz, Chloroform-d) δ 5.30 (s, 2H), 3.53 (t, J = 7.9 Hz, 2H), 2.55 (s, 3H), (m, 1H), (m, 6H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 142.8, 130.7, 114.4, 72.7, 66.2, 17.7, 16.1, 5.6, 4.3, LRMS (ESI-MS): 363.0/365.0 [M+H] + 5-bromo-4-cyclobutyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55h): The title compound was prepared from 54h according to general Procedure O and obtained as colorless sticky oil after gradient silica flash chromatography (hexane hexane/etoac 9:1) in 94% yield. 1 H NMR (200 MHz, Chloroform-d) δ 5.24 (s, 2H), (m, 3H), 2.52 (s, 3H), (m, 6H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 143.9, 143.1, 99.7, 73.4, 66.2, 32.8, 27.9, 18.5, 17.7, 17.1, LRMS (ESI-MS): 377.0/379.0 [M+H] + 5-bromo-4-cyclopentyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55i): The title compound was prepared from 54i according to general Procedure O and obtained as colorless sticky oil after gradient silica flash chromatography (hexane hexane/etoac 6:1) in 85% yield. 1 H NMR (200 MHz, Chloroform-d) δ 5.30 (d, J = 1.7 Hz, 2H), (m, 2H), (m, 1H), 2.54 (s, 3H), (m, 8H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 144.5, 142.9, 100.2, 73.5, 66.2, 37.8, 32.0, 25.5, 17.7, 17.2, LRMS (ESI-MS): 391.1/393.1 [M+H] + 5-bromo-4-(tert-butyl)-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (55j): The title compound was prepared from 54j according to general Procedure O and obtained as brown sticky oil in 80% yield, which was used without further purification. 1 H NMR (200 MHz, 30

31 Chloroform-d) δ 5.31 (s, 2H), (m, 2H), 2.56 (s, 3H), 1.38 (s, 9H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 147.7, 141.6, 98.2, 73.4, 66.2, 32.6, 29.5, 17.7, 17.1, LRMS (ESI-MS): 379.1/381.1 [M+H] + 2-(2,5-dimethyl-1H-pyrrol-1-yl)-4-(2-(methylthio)-4-(naphthalen-2-yl)-1-((2- (trimethylsilyl)ethoxy)methyl) -1H-imidazol-5-yl)pyridine (56c): The title compound was prepared from 55c and 2-(2,5-dimethyl-1H-pyrrol-1-yl)-4-(4,4,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine according to general Procedure P (method A) and obtained as colorless sticky oil after gradient silica flash chromatography(hexane hexane/etoac 6:1) in 70% yield. 1 H NMR (200 MHz, Chloroform-d) δ 8.66 (d, J = 5.1 Hz, 1H), (m, 1H), (m, 3H), (m, 4H), (m, 1H), 5.83 (s, 2H), 5.26 (s, 2H), (m, 2H), 2.82 (s, 3H), 2.01 (s, 6H), 1.00 (d, J = 8.4 Hz, 2H), 0.04 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 152.9, 150.0, 146.7, 141.3, 140.7, 133.5, 132.8, 131.0, 128.7, 128.2, 128.1, 127.7, 127.6, 126.8, 126.2, 126.0, 125.8, 123.5, 123.3, 107.0, 73.2, 66.9, 18.1, 16.4, 13.2, LRMS (ESI-MS): [M+H] + N-(4-(2-(methylthio)-4-(thiophen-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)pyridin-2-yl)acetamide (56d): The title compound was prepared from 55d and N-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2- yl)pyridin-2-yl)acetamide according to general Procedure P (method A) and obtained as yellow sticky oil after gradient silica flash chromatography(hexane hexane/etoac 3:1) in 76% yield. 1 H NMR (200 MHz, Chloroform-d) δ 9.38 (s, 1H), (m, 2H), (m, 4H), 5.14 (s, 2H), (m, 2H), 2.67 (s, 3H), 2.15 (s, 3H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 169.0, 152.1, 147.6, 145.8, 145.7, 140.4, 136.4, 135.1, 127.0, 31

32 126.6, 124.4, 123.9, 121.3, 120.2, 115.5, 74.9, 72.9, 66.3, 24.7, LRMS (ESI-MS): [M+H] + 2-(2,5-dimethyl-1H-pyrrol-1-yl)-4-(2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1Himidazol-5-yl)pyridine (56e): 1H), (m, 4H), (m, 2H), 2.70 (s, 6H), 2.20 (s, 3H), (m, 2H), (m, 9H). LRMS (ESI-MS): [M+H] + tert-butyl The title compound was prepared from 55e and N-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2- yl)pyridin-2-yl)acetamide according to general Procedure P (method A) and obtained a brownish sticky oil after gradient silica flash chromatography(hexane hexane/etoac 2:1) in 54% yield. 1 H NMR (200 MHz, Chloroform-d) δ (m, 1H), (m, 2H), (m, (4-(4-methyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)pyridin-2-yl)carbamate (56f): The title compound was prepared from 55f and tert-butyl (4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)pyridin-2-yl)carbamate according to general Procedure P (method B) and obtained as a yellow semi-solid after gradient silica flash chromatography(dcm DCM/iPrOH 5%) in 81% yield. 1 H NMR (200 MHz, Chloroform-d) δ 9.54 (s, 1H), 8.36 (d, J = 5.4 Hz, 1H), 8.28 (s, 1H), 7.42 (d, J = 5.2 Hz, 1H), 5.33 (s, 2H), (m, 2H), 2.68 (s, 3H), 2.57 (s, 3H), 1.58 (s, 9H), (m, 2H), 0.03 (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 171.0, 152.7, 147.6, 144.5, 143.0, 136.2, 128.6, 116.1, 109.4, 80.4, 72.8, 66.1, 28.3, 24.8, 17.7, 10.5, LRMS (ESI-MS): [M+H] + 32

33 N-(4-(4-cyclopropyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)pyridin-2-yl)acetamide (56g): The title compound was prepared from 55g and N-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2- yl)pyridin-2-yl)acetamide according to general Procedure P (method B) and obtained as a white solid after gradient silica flash chromatography(hexane hexane/etoac 2:1) in 84% yield. 1 H NMR (200 MHz, Chloroform-d) δ 9.74 (s, 1H), (m, 2H), 7.20 (s, 1H), 5.13 (s, 2H), 3.46 (t, J = 8.5 Hz, 2H), 2.49 (s, 3H), 2.12 (s, 3H), (m, 1H), (m, 6H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 169.2, 152.1, 147.3, 145.2, 143.7, 140.5, 128.0, 119.5, 114.2, 74.9, 73.0, 66.1, 24.8, 17.7, 16.5, 8.5, 7.4, LRMS (ESI-MS): [M+H] + N-(4-(4-cyclobutyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)pyridin-2-yl)acetamide (56h): The title compound was prepared from 55h and N-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2- yl)pyridin-2-yl)acetamide according to general Procedure P (method A) and obtained as a white solid after gradient silica flash chromatography(hexane hexane/etoac 1:1) in 81% yield. 1 H NMR (200 MHz, Chloroform-d) δ 9.33 (s, 1H), 8.28 (d, J = 5.3 Hz, 1H), 8.22 (s, 1H), (m, 1H), 5.22 (s, 2H), (m, 3H), 2.67 (s, 3H), (m, 2H), (m, 5H), (m, 2H), (m, 2H), (s, 9H). 13 C NMR (50 MHz, Chloroform-d) δ 169.0, 152.1, 147.5, 145.7, 145.3, 140.6, 127.2, 119.9, 114.4, 73.2, 66.4, 33.1, 28.8, 24.8, 18.6, 17.9, 17.1, LRMS (ESI-MS): [M+H] + tert-butyl (4-(4-cyclopentyl-2-(methylthio)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)pyridin-2-yl)carbamate (56i): 33