Supplementary Material. Synthesis, Biological Evaluation, and Structure-Activity Relationships of Potent Noncovalent

Transcription

1 Supplementary Material Synthesis, Biological Evaluation, and Structure-Activity Relationships of Potent Noncovalent and Nonpeptidic Cruzain Inhibitors as Anti-Trypanosoma cruzi Agents Rafaela S. Ferreira, ǁ, Marco A. Dessoy, ǁ, Ivani Pauli, Mariana L. Souza, Renata Krogh, Ana I. L. Sales, Glaucius Oliva, Luiz C. Dias,*, Adriano D. Andricopulo*, Table of contents SYNTHETIC PROCEDURES AND COMPOUND CHARACTERIZATION REFERENCES S2 S32 1

2 SYNTHETIC PROCEDURES AND COMPOUND CHARACTERIZATION General Procedures for Preparation of Compounds of Type 8 Method A: The matching 2-phenoxyacetic acid (6, 15 mmol) was dissolved in absolute DCM (50 ml) containing dry Et 3 N (22.5 mmol) at 0 C under an Ar atmosphere. Neat PivCl (15 mmol) was added drop wise within 15 min, and a white precipitate formed and was separated. The slurry was stirred for 20 min, the ice-bath was removed, and then stirring was continued for 1 h. The mixture containing the crude mixed anhydride was stored at 0 C. In a separate reaction flask, commercial 2-(2-aminoethyl)benzimidazole dihydrochloride (7, 0.20 g, 0.85 mmol) was suspended in absolute THF under an Ar atmosphere. Dry Et 3 N (3.06 mmol) was added, and the mixture stirred until complete dissolution of 2-dihydrochloride occurred (eventually, gentle warming was required). The THF solution was cooled down to -78 C, and the cold DCM solution containing the crude mixed anhydride (3 ml) was added portionwise within 15 min. Stirring was continued for 2 h at -78 C. The reaction mixture was allowed to warm up to room temperature, and the solvent blend was evaporated under reduced pressure. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (three times); diluted NaHCO 3 solution (two times) and brine; dried over Na 2 SO 4 ; and concentrated to dryness in vacuum. The crude solid product was suspended in diethyl ether, and the resulting slurry was stirred for 1 h at room temperature. The precipitate was filtered off, rinsed with diethyl ether, collected, and dried in vacuum. 2

3 Method B: The matching 2-phenoxyacetic acid (1.5 mmol) was suspended in absolute DCM (15 ml) at room temperature. A catalytic amount of N,N-dimethylformamide (DMF, 0.10 ml) was added, and then neat oxalyl chloride (0.63 g, 5 mmol) was combined portionwise within 10 min. Stirring was continued until gas evolution ceased. The clear solution that resulted was cooled down to 0 C and combined with solid N-hydroxysuccinimide (0.69 g, 6 mmol), and then neat dry Et 3 N (1.52 g, 15 mmol) was added portionwise within 10 min. After 15 min, the mixture was removed from the ice bath, and stirring was continued for 2 h. The solvent was evaporated in vacuum, and the solid evaporation residue was partitioned between ethyl acetate and ice-cold water. The organic layer was washed with cold diluted NaHCO 3 solution and brine, dried over Na 2 SO 4, and concentrated in vacuum. The crude N-hydroxysuccinimide ester thus obtained was suspended in diethyl ether (3 ml), and the mixture was stirred at room temperature (15 min). Hexane (3 ml) was added, and the mixture was kept in a freezer overnight. The solid was filtered off, rinsed with cold diethyl ether, collected, dried in vacuum, and stored. In a separate reaction flask, commercial 2-(2-aminoethyl)benzimidazole dihydrochloride (7, 0.25 g, 1.07 mmol) was suspended in ordinary DCM (30 ml). Neat, ordinary Et 3 N (0.36 g, 3.52 mmol) was added, and the matching solid N-hydroxysuccinimide ester was added to the clear solution that resulted. The mixture was stirred at room temperature for 15 min. The solvent was evaporated in vacuum, and the evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with diluted NaOH solution and brine, dried over Na 2 SO 4, and concentrated to dryness in vacuum. The crude solid product was suspended in diethyl ether, and the resulting slurry was stirred for 1 h at room temperature. The precipitate was filtered off, rinsed with diethyl ether, collected, and dried in vacuum. Unless otherwise stated, no further purification was required. 3

4 N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-phenoxyacetamide (8a, method B): 91% yield, pale, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.36 (t, 1H, J ~ 5.3 Hz), (m, 2H), (m, 2H), (m, 2H), (m, 3H), 4.47 (s, 2H), 3.61 (q, 2H, J ~ 6.5 Hz), 3.43 (t, 2H, J ~ 7.0 Hz); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , 66.94, 36.83, 28.61; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 18 O 2 N , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-fluorophenoxy)acetamide (8b, method B): 92% yield, white, amorphous solid; 1 H NMR (500 MHz, DMSO-d 6 ) (bs, 1H), 8.34 (t, 1H,, J = 5.7 Hz), (m, 2H), (m, 1H), (m, 2H), (m, 2H), (m, 1H), 4.55 (s, 2H), 3.61 (q, 2H, J ~ 7.0 Hz), 3.00 (t, 2H, J ~ 7.0 Hz); ); 13 C NMR (125 MHz, DMSO-d 6 ) , , (d, J C,F = 244 Hz), (d, J C,F = 10 Hz), , , (d, J C,F = 7 Hz), , (d, J C,F = 18 Hz), , 67.75, 36.82, 28.54; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 N 3 O 2 F , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-chlorophenoxy)acetamide (8c, method B): 94% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.20 (t, 1H,, J = 5.5 Hz), (m, 1H), (m, 2H), (m, 3H), (m, 2H), 4.58 (s, 2H), 3.65 (q, 2H, J ~ 6.6 Hz), 3.01 (t, 2H, J ~ 6.6 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , 67.57, 36.60, 28.52; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 N 3 O 2 Cl , found

5 N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-bromophenoxy)acetamide (8d, method A): 27% yield, colorless, microcrystalline solid (after re-crystallization from hot methanol); 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.10 (bt, 1H, J ~ 5.8 Hz), 7.53 (dd, 1H, J = 1.6, 7.9 Hz), (m, 2H), 7.24 (ddd, 1H, J = 1.6, 7.3, 8.2 Hz), (m, 2H), 6.96 (dd, 1H, J = 1.3, 8.2 Hz), 6.87 (dt, 1H, J = 1.4, 7.6 Hz), 4.57 (s, 2H), 3.66 (q, 1H, J = 6.4 Hz), 3.01 (t, 1H, J = 6.4 Hz); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , 67.67, 36.52, 28.56; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 BrO 2 N , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-iodophenoxy)acetamide (8e, method B): 90% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 7.99 (t, 1H, J ~ 5.3 Hz), 7.72 (d, 1H, J = 7.6 Hz), (m, 2H), 7.25 (t, 1H, J = 7.7 Hz), (m, 2H), 6.87 (d, 1H, J = 8.2 Hz), 6.73 (t, 1H, J = 7.5 Hz), 4.54 (s, 2H), 3.69 (q, 2H, J ~ 6.3 Hz), 3.02 (t, 2H, J ~ 6.5 Hz); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , , , , 86.30, 67.87, 36.56, 28.63; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 IN 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-cyanophenoxy)acetamide (8f, method B): 95% yield, white powder; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.33 (t, 1H, J ~ 5.4 Hz), 7.70 (dd, 1H, J = 1.6, 7.6 Hz), (m, 3H), (m, 4H), 4.70 (s, 2H), 3.59 (q, 2H, J ~ 6.5 Hz), 2.98 (t, 2H, J ~ 6.9 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , 67.28, 36.95, 28.58; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 17 N 4 O , found

6 N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-nitrophenoxy)acetamide (8g, method B): 79% yield, white, amorphous solid; 1 H NMR (500 MHz, DMSO-d 6 ) (bs, 1H), 8.82 (t, 1H, J = 5.7 Hz), 7.87 (dd, 1H, J = 1.7, 8.1 Hz), 7.54 (ddd, 1H, J = 1.7, 7.3, 8.5 Hz), (m, 2H), 7.18 (dd, 1H, J = 1.2, 8.5 Hz), (m, 3H), 4.71 (s, 2H), 3.61 (q, 2H, J ~ 7.1 Hz), 2.99 (t, 2H, J ~ 7.1 Hz); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , 67.71, 36.87, 28.58; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 O 4 N , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-aminophenoxy)acetamide (8h): Compound 8g (0.48 g, 1.4 mmol) was dissolved in hot ethanol (30 ml), and the solution was cooled down to room temperature, and then acidified with 1 M aq. HCl solution. 5% palladium on charcoal (50 mg) was added and the mixture was vigorously stirred at room temperature for 3 h. The catalyst was filtered off (celite pad), and the solution was concentrated in vacuum. The evaporation residue was diluted with water (10 ml), alkalinized with Na 2 CO 3, and extracted with ethyl acetate (3 times). The combined organic phases were washed with brine. Na 2 SO 4 and silica gel (ca. 1g) were added and the slurry was stirred for 10 min. The solid material was filtered off, and the clear solution concentrated to dryness in vacuum. The crude product was suspended in diethyl ether (5 ml), and the slurry was stirred at room temperature for 1 h. The finely divided solid was filtered off, rinsed with diethyl ether, and dried in vacuum g (54% yield), pale powder; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.48 (t, 1H, J = 5.6 Hz), (m, 2H), (m, 2H), (m, 3H), 6.46 (m, 1H), 5.08 (s, 2H), 4.40 (s, 2H), 3.64 (q, 2H, J ~ 6.7 Hz), 3.04 (t, 2H, J = 7.1 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , 114,13, , , 6

7 67.21, 36.93, 28.71; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 19 N 4 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(2-acetamidophenoxy)acetamide (8i): Compound 8h (0.25 g, 0.80 mmol) was suspended in a mixture of absolute THF (20 ml) and dry pyridine (0.26 ml, 3.2 mmol) at 0 C. Neat acetic anhydride (0.18 ml, 1.76 mmol) was added, the icebath was then removed after 15 min, and the mixture was stirred at room temperature overnight. Ordinary methanol was added until dissolution of the resulting precipitated, and the mixture was stirred at 40 o C for 24 h. The solvent blend was evaporated in vacuum and the evaporation residue portioned between water and ethyl acetate. The organic layer was washed with water (once), diluted NaHCO 3 solution (three times), brine, dried over MgSO 4, and concentrated to dryness in vacuum. The crude product was suspended in a 1:2 v/v blend of ethyl acetate and diethyl ether and stirred at room temperature for 1 h. The slurry was kept in a freezer overnight. The solid was filtered off, rinsed with cold ethyl acetate, and dried in vacuum. 183 mg (64% yield), white powder; 1 H NMR (500 MHz, DMSO-d 6 ) (bs, 1H), 9.38 (s, 1H), 8.49 (t, 1H, J ~ 6 Hz), 7.84 (d, 1H, J = 7.6 Hz), (m, 2H), (m, 2H), (m, 2H), 6.92 (m, 1H), 4.53 (s, 2H), 3.62 (dd, 2H, J = 7.1 Hz, J ~ 6 Hz), 3.02 (t, 2H, J = 7.3 Hz), 2.02 (s, 3H); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , , 67.75, 37.06, 28.72, 23.75; HRMS (ES+) m/z ([M + H] + ) calculated for C 19 H 21 N 4 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(o-tolyloxy)acetamide (8j, method A): 30% yield, colorless, microcrystalline solid (after re-crystallization from hot methanol); 1 H NMR (250 MHz, 7

8 DMSO-d 6 ) (bs, 1H), 8.21 (bt, 1H, J ~ 5.3 Hz), (m, 2H), (m, 4H), (m, 2H), 4.47 (s, 2H), 3,64 (q, 2H, J = 6.5 Hz), 3.00 (t, 2H, J = 6.5 Hz), 2.20 (s, 3H); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , 67.04, 36.55, 28.49, 16.13; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 20 O 2 N , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(3-bromophenoxy)acetamide (8k, method B): 85% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.38 (t, 1H, J ~ 5.5 Hz), (m, 2H), (m, 5H), (m, 1H), 4.51 (s, 2H), 3.60 (q, 2H, J ~ 6.6 Hz), 3.00 (t, 2H, J ~ 7.1 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , 67.06, 36.86, 28.60; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 BrN 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(4-bromophenoxy)acetamide (8l, method B): 91% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.36 (t, 1H, J ~ 5.6 Hz), (m, 2H), 7.39 (d, 2H, J = 9.0 Hz), (m, 2H), 6.88 (d, 2H, J = 9.0 Hz), 4.47 (s, 2H), 3.59 (q, 2H, J ~ 6.7 Hz), 2.99 (t, 2H, J ~ 7.0 Hz); 13 C NMR (60 MHz, DMSOd 6 ) , , , , , , , 67.14, 36.91, 28.62; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 17 BrN 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(naphthalen-1-yloxy)acetamide (8m, method B): 78% yield, pale, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), (t, 1H, J ~ 5.6 Hz), 8.34 (d, 1H, J = 8.2 Hz), 7.86 (d, 1H, J = 8.1 Hz), (m, 5H), 7.36 (t, 1H, J 8

9 = 7.9 Hz), (m, 2H), 6.90 (d, 1H, J = 7.4 Hz), 4.68 (s, 2H), 3.70 (q, 2H, J ~ 6.4 Hz), 3.07 (t, 2H, J = 6.8 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , 67.29, 36.70, 28.52; HRMS (ES+) m/z ([M + H] + ) calculated for C 21 H 20 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(naphthalen-2-yloxy)acetamide (8n, method B): 94% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), (t, 1H, J = 5.5 Hz), (m, 2H), 7.75 (d, 1H, J = 8.0 Hz), (m, 6H), (m, 2H), 4.60 (s, 2H), 3.67 (q, 2H, J ~ 6.6 Hz), 3.03 (t, 2H, J = 7.1 Hz); 13 C NMR (60 MHz, DMSOd 6 ) , , , , , , , , , , , , , , , , , 67.04, 36.88, 28.63; HRMS (ES+) m/z ([M + H] + ) calculated for C 21 H 20 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(1-formylnaphthalen-2-yloxy)acetamide (8o, method B): 34% yield, pale, amorphous solid; 1 H NMR (500 MHz, DMSO-d 6 ) (bs, 1H), (s, 1H), 9.14 (d, 1H, J = 8.5 Hz), 8.50 (s, 1H), 8.16 (d, 1H, J = 9.0 Hz), 7.91 d, 1H, J = 7.8 Hz), 7.64 (t, 1H, J ~ 7 Hz), (m, 4H), 7.12 (s, 2H), 4.84 (s, 2H), 3.64 (m, 2H), 3.03 (m, 2H); 13 C NMR (100 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , 67.99, 36.99, 28.57; HRMS (ES+) m/z ([M + H] + ) calculated for C 22 H 20 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(1-(hydroxymethyl)naphthalen-2-yloxy)acetamide (8p): Compound 8o (350 mg, 0.94 mmol) was dissolved in hot ethanol (20 ml), and the solution 9

10 was cooled to 0 C to receive the addition of solid sodium borohydride (0.19 g, 5.0 mmol). The ice-bath was removed, and then stirring was continued for 2 h. The reaction mixture was cooled to 0 C, and the excess of the reducing agent was destroyed with acetic acid. The solvent was evaporated in vacuum, and the evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (once), brine, dried over Na 2 SO 4, and concentrated to dryness in vacuum. The crude product was suspended in ethanol, and the resulting slurry was stirred for 1 h at room temperature, and then kept in a freezer overnight. The solid material was filtered off, rinsed with cold ethanol, and dried in vacuum. 150 mg (40% yield), white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.39 (bs, 1H), 8.16 (d, 1H, J = 8.4 Hz), 7.82 (m, 2H), (m, 5H), (m, 2H), 5.10 (s, 1H), 5.01 (s, 2H), 4.66 (s, 2H), (m, 2H), 3.02 (t, 2H, J = 6.7 Hz); 13 C NMR (60 MHz, DMSOd 6 ) , , , , , , , , , , , , , 68.68, 53.37, 36.93, 28.62; HRMS (ES+) m/z ([M + H] + ) calculated for C 22 H 22 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(1-hydroxynaphthalen-2-yloxy)acetamide (8q): Compound 8o (250 mg, 0.67 mmol) was dissolved in hot ethanol (15 ml), and the solution was cooled to room temperature, and then solid 3-chloroperbenzoic acid (0.3 g, ca. 77 % purity) was added. The mixture was stirred for 12 h. An aqueous solution of Na 2 SO 3 (1g in 10 ml water) was added, the solution was stirred for 1 h, and then concentrated in vacuum. The evaporation residue was diluted with saturated NaHCO 3 solution (10 ml) and extracted with ethyl acetate (three times). The combined organic phases were washed with brine, dried over Na 2 SO 4, and concentrated to dryness in vacuum. The crude product was suspended and stirred for 1 h in a 10

11 blend of ethanol and ethyl acetate (5 ml, 1:1, v/v). The slurry was kept in a fridge overnight, the solid material filtered off, rinsed with ethyl acetate, and dried in vacuum. 150 mg (37%), pale powder; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 9.68 (bs, 1H), 8.71 (t, 1H, J ~ 5 Hz), 8.10 (d, 1H, J = 8.2 Hz), 7.77 (d, 1H, J = 8.0 Hz), (m, 6H), (m, 2H), 4.58 (s, 2H), 3.68 (q, 2H, J ~ 6 Hz), 3.07 (t, 2H, J ~ 7 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , 69.09, 37.11, 28.84; HRMS (ES+) m/z ([M + H] + ) calculated for C 21 H 20 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(1-bromonaphthalen-2-yloxy)acetamide (8r, method B): 65% yield, white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.20 (bt, 1H, J ~ 5.5 Hz), 8.05 (d, 1H, J = 8.5 Hz), 7.88 (m, 2H), (m, 5H), (m, 2H), 4.73 (s, 2H), 3.69 (q, 2H, J ~ 6.0 Hz), 3.04 (t, 2H, J = 6.6 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , , , , , 68.32, 36.61, 28.58; HRMS (ES+) m/z ([M + H] + ) calculated for C 21 H 19 O 2 N 3 Br , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-3-(2-bromophenyl)propanamide (8s, method A): 42% yield, colorless, crystalline solid; 1 H NMR (500 MHz, DMSO-d 6 ) (s, 1H), 8.06 (t, 1H, J = 5.7 Hz), 7.54 (dd, 1H, J = 1.0, 8.0 Hz), 7.52 (bd, 1H, J = 6.9 Hz), 7.40 (bd, 1H, J = 6.7 Hz), 7.28 (dd, 1H, J = 1.8, 7.8 Hz), 7.24 (dt, 1H, J = 1.2, 7.3 Hz), (m, 3H), 3.50 (q, 2H, J = 6.7 H Hz), 2.93 (t, 2H, J = 7.3 Hz), 2.90 (t, 2H, J = 7.8 Hz), 2.37 (m, 2H); ); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , , 11

12 123.63, , , 118,11, , 37.20, 35.04, 31.29, 28.91; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 19 N 3 OBr , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(phenylthio)acetamide (8t, method B): Purification: crude product was suspended in DCM (3 ml), and then diluted with diethyl ether (4 ml). The slurry was kept in a freezer overnight. Solid was filtered off, rinsed with diethyl ether, collected, and dried in vacuum. 92% yield, pale microneedles; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.33 (t, 1H, J = 5.5 Hz), (m, 2H), (m, 7H), 3.63 (s, 2H), 3.53 (q, 2H, J ~ 6.8 Hz), 2.93 (t, 2H, J ~ 6.8 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , 37.41, 36.40, 28.56; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 18 N 3 OS , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)acetamide (8u): Commercial 2-(2- aminoethyl)benzimidazole dihydrochloride (7, 0.10 g, 0.42 mmol) was suspended in absolute DCM (4 ml) and acetic anhydride (1 ml). The mixture was cooled to 0 o C, neat, dry Et 3 N (0.69 ml, 5.4 mmol) was combined, and the mixture was stirred for 1 h at room temperature. The reaction mixture was cooled to 0 o C, methanol (1 ml) was added, and the mixture was stirred for 1 h at room temperature. The reaction solution was concentrated in vacuum. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with saturated NaHCO 3 solution (three times), dried over Na 2 SO 4, and concentrated in vacuum. The crude product was suspended in diethyl ether (3 ml), stirred for 1 h at room temperature, and then kept in a freezer overnight. The solid was filtered off, rinsed with diethyl ether, collected, and dried in vacuum. 55 mg, 64% yield, pale, crystalline solid; 1 H NMR (250 MHz, CD3OD) 12

13 7.49 (dd, 2H, J = 3.1, 6.0 Hz), 7.18 (dd, 2H, J = 3.1, 6.0 Hz), 3.61 (t, 2H, J = 7.0 Hz), 3.06 (t, 2H, J = 7.0 Hz), 1.90 (s, 3H); 13 C NMR (125 MHz, CD3OD) , , , , , 39.12, 29.98, 22,51; HRMS (ES+) m/z ([M + H] + ) calculated for C 11 H 14 N 3 O , found N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-3-phenoxypropanamide (8v, method A): 38% yield, white powder; 1 H NMR (250 MHz, CD3OD) (m, 2H), (m, 4H), 6.87 (t, 1H, J = 7.3 Hz), 6.79 (d, 2H, J = 8.2 Hz), 4.16 (t, 2H, J = 6.1 Hz), 3.68 (t, 2H, J = 6.9 Hz), 3.10 (t, 2H, J = 6.9 Hz), 2.60 (t, 2H, J = 6.1 Hz); 13 C NMR (60 MHz, CD3OD) , , , , , , , , , 65.11, 38.96, 37.16, 29.90; HRMS (ES+) m/z ([M + H] + ) calculated for C 19 H 22 N 2 O , found General Procedures for Preparation of Compounds of Type 10 Method A: The matching 2-phenoxyacetic acid (6, 15 mmol) was dissolved in absolute DCM (50 ml) containing dry Et 3 N (22.5 mmol) at 0 C under an Ar atmosphere. Neat PivCl (15 mmol) was added dropwise over 15 min, whereupon a white precipitate separated. The slurry was stirred for 20 min, the ice bath was removed, and stirring was continued for 1 h. The mixture containing the crude mixed anhydride was stored at 0 C. In a separate reaction flask, the matching amine of type 9 (2 mmol) was dissolved in absolute THF (10 ml). Neat, dry Et 3 N (0.20 g, 2 mmol) was added. The mixture was cooled down to -78 C, and then the cold DCM solution containing the crude mixed anhydride (12 ml, ca. 3.6 mmol) was added portionwise within 10 min. The mixture was stirred for 2 h. The reaction mixture was concentrated in 13

14 vacuum, and the evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (once), saturated NaHCO 3 solution (twice) and brine; dried over Na 2 SO 4 ; and concentrated to dryness in vacuum to give the crude products. Method B: The matching 2-phenoxyacetic acid (6, 5 mmol) was suspended in absolute DCM (15 ml) at room temperature. A catalytic amount of DMF (0.10 ml) was added, and then neat oxalyl chloride (0.63 g, 5 mmol) was combined portionwise within 10 min. Stirring continued until gas evolution ceased. The clear solution that resulted was cooled down to 0 C, solid N- hydroxysuccinimide (0.69 g, 6 mmol) was combined, and then neat dry Et 3 N (1.52 g, 15 mmol) was added portionwise within 10 min. After 15 min, the mixture was removed from the ice bath, and stirring was continued for 2 h. The solvent was evaporated in vacuum, and the solid evaporation residue was partitioned between ethyl acetate and ice-cold water. The organic layer was washed with cold diluted NaHCO 3 solution and brine, dried over Na 2 SO 4, and concentrated in vacuum. The crude N-hydroxysuccinimide ester thus obtained was suspended in diethyl ether (3 ml), and the mixture was stirred at room temperature (15 min). Hexane (3 ml) was added, and the mixture was kept in a freezer overnight. The solid was filtered off, rinsed with cold diethyl ether, collected, dried in vacuum, and stored. In a separate reaction flask, commercial 2- (1H-imidazol-4-yl)ethanamine dihydrochloride (9D, g, 1.06 mmol) was suspended in ordinary ethyl acetate (40 ml). Neat, ordinary Et 3 N (0.21 g, 2.12 mmol) was added, and the mixture was stirred for 30 min at room temperature. The matching solid N-hydroxysuccinimide ester was added, and the mixture was stirred at room temperature for 15 min. The reaction solution was washed with water and then extracted with 2 M HCl solution. The organic phase was discarded. The aqueous phase was alkalinized using 2 M NaOH solution and extracted with 14

15 ethyl acetate (three times). The combined organic phases were washed with brine, dried over Na 2 SO 4, and concentrated to dryness in vacuum. The products were obtained in pure form, and no purification was required. N-(2-(1H-imidazol-5-yl)ethyl)-2-(2-chlorophenoxy)acetamide (10cD, method B): g (53% yield), white, amorphous solid; 1 H NMR (500 MHz, CDCl 3 ) 7.64 (s, 1H), (m, 2H), 7.19 (m, 1H), 6.93 (m, 1H), (m, 2H), 6.69 (bs, 2H), 4.48 (s, 2H), 3.66 (q, 2H, J = 6.4 Hz), 2.86 (t, 2H, J = 6.6 Hz); 13 C NMR (125 MHz, CDCl 3 ) , , , , , , , , , , 67.89, 38.56, 26.91; HRMS (ES+) m/z ([M + H] + ) calculated for C 13 H 15 ClN 3 O , found (2-bromophenoxy)-N-(2-(piperidin-1-yl)ethyl)acetamide (10dB, method A): The crude product was dissolved in diethyl ether and extracted with 1 M aqueous HCl solution (10 ml). The acidic aqueous phase was washed with diethyl ether and both ethereal phases were discarded. The acidic aqueous phase was alkalinized with aqueous 2 M NaOH solution and reextracted (three times) with diethyl ether. The combined ethereal phases were washed with brine, dried over Na 2 SO 4, concentrated in vacuum, and dried under high vacuum. This simple procedure rendered the products in very good purity. 94% yield, slightly colored oil, 1 H NMR (250 MHz, CDCl 3 ) 7.55 (dd, 1H, J = 1.6, 7.9 Hz), 7.45 (bs, 1H), 7.27 (dt, 1H, J = 1.58, 7.8 Hz), 6.90 (dd, 1H, J = 1.3, 7.7 Hz), 6.84 (dd, 1H, J = 1.3, 8.2 Hz), 4.51 (s, 2H), 3,42 (q, 2H, J = 5.8 Hz), 2.45 (t, 2H, J = 5.8 Hz), (m, 4H), (m, 6H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , 67.73, 56.97, 54.29, 35.78, 15

16 25.99, 24.36; HRMS (ES+) m/z ([M + H] + ) calculated for C 15 H 22 BrO 2 N , found N-(2-(piperidin-1-yl)ethyl)-2-(o-tolyloxy)acetamide (10jB, method A): The crude product was purified as described for 10dB. 64% yield, slightly colored syrup; 1 H NMR (250 MHz, CDCl 3 ) 7.36 (bs, 1H), (m, 2H), 6.90 (m, 1H), 6.75 (m, 1H), 4.48 (s, 2H), 3.39 (q, 2H, J = 5.5 Hz), 2.43 (t, 2H, J = 5.5 Hz), (m, 9H), (m, 6H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , 121,47, , 67.05, 56.79, 54.18, 35.40, 25.93, 24.33, 16.46; HRMS (ES+) m/z ([M + H] + ) calculated for C 16 H 25 O 2 N , found N-(2-hydroxyethyl)-2-(o-tolyloxy)acetamide (10sB, method A): 1 H NMR (250 MHz, CDCl 3 ) (m, 2H), 7.05 (bs, 1H), 6.92 (t, 1H, J = 7.1 Hz ), 6.75 (d, 2H, J = 8.5 Hz), 4.49 (s, 2H), 3.75 (t, 2H, J = 5.0 Hz), 3.51 (q, 2H, J = 5.0 Hz), 2.72 (bs, 1H), 2.27 (s, 3H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , 67.46, 61.90, 41.78, 16.27; IR ; HRMS (ES+) m/z ([M + H] + ) calculated for C 11 H 16 NO , found (2-bromophenyl)-N-(2-(piperidin-1-yl)ethyl)propanamide (10jA, method A): The crude product was suspended, stirred in diethyl ether, kept in a freezer overnight, filtered, and then dried in vacuum. 40% yield, colorless microcrystalline solid; 1 H NMR (250 MHz, CDCl 3 ) 7.50 (d, 1H, J = Hz), (m, 2H), 7.03 (dt, 1H, J = 2.0, 7.4 Hz), 6.11 (bs, 1H), 3.26 (q, 2H, J = 5.7 Hz), 3.06 (t, 2H, J = 7.7 Hz), 2.47 (t, 2H, J = 7.7 Hz), (m, 6H), (m, 6H); ); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , 16

17 124.23, 57.00, 54.14, 36.43, 35.92, 32.23, 25.91, 24.29; HRMS (ES+) m/z ([M + H] + ) calculated for C 16 H 24 N 2 OBr , found N-(2-(1H-indol-3-yl)ethyl)-2-(o-tolyloxy)acetamide (10jC, method A): The crude product was recrystallized from diethyl ether (-20 to -78 C); 1 H NMR (250 MHz, CDCl 3 ) 8.13 (bs, 1H), 7.60 (d, 1H, J = 7.9 Hz), 7.35 (d, 1H, J = 8.1 Hz), (m, 4H), 6.90 (m, 2H), (m, 2H), 4.45 (s, 2H), 3.71 (q, 2H, J = 6.4 Hz), 3.01 (t, 2H, J = 6.4 Hz), 2.01 (s, 3H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , , , , , , , , 67.33, 38.87, 25.20, N-(2-(1H-indol-3-yl)ethyl)-2-(1-bromonaphthalen-2-yloxy)acetamide (10rC, method A): The crude product was suspended, and stirred in diethyl ether, kept in a fridge overnight, solid material filtered off, rinsed with diethyl ether, and then dried in vacuum. 79% yield, pale microcrystalline solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.08 (d, 1H, J = 8.5 Hz), (m, 3H), 7.63 (m, 1H), 7.56 (d, 1H, J = 7.7 Hz), 7.46 (m, 1H), 7.35 (d, 2H, J = 8.8 Hz), 7.19 (d, 1H, J = 2.0 Hz), 7.06 (m, 1H), 6.97 (m, 1H), 4.72 (s, 2H), 3.50 (q, 2H, J = 6.7 Hz), 2.90 (t, 2H, J = 7.2 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , , , , 68.45, 39.09, 25.02; HRMS (ES+) m/z ([M + H] + ) calculated for C 22 H 20 O 2 N 2 Br , found N-(2-(1H-indol-3-yl)ethyl)-3-phenoxypropanamide (10vC, method A): The crude product was suspended, and stirred in ethyl acetate (2 ml), diethyl ether (5 ml) was added and stirring 17

18 continued for 1 h. The slurry was kept in a freezer overnight, the solid material was filtered off, rinsed with diethyl ether, and then dried in vacuum. 78% yield, white, amorphous solid; 1 H NMR (250 MHz, CDCl 3 ) 8.04 (bs, 1H), 7.59 (d, 1H, J = 7.7 Hz), 7.34 (d, 1H, J = 8.0 Hz), (m, 3H), 7.11 (t, 1H, J = 7.4 Hz), 6.94 (m, 2H), 6.77 (d, 2H, J = 7.9 Hz), 6.02 (bs, 1H), 4.18 (t, 2H, J = 5.9 Hz), 3.62 (q, 2H, J = 6.4 Hz), 2.97 (t, 2H, J = 6.4 Hz), 2.59 (t, 2H, J = 5.9 Hz); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , , , , , , , 64.09, 39.70, 36.68, 25.19; HRMS (ES+) m/z ([M + H] + ) calculated for C 19 H 21 O 2 N , found N-(2-(1H-imidazol-5-yl)ethyl)-2-(naphthalen-2-yloxy)acetamide (10nD, method B): g (46% yield), white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) 8.31 (bt, 1H, J = 5.2 Hz), (m, 3H), 7.70 (s, 1H), 7.45 (m, 1H), 7.35 (m, 1H), (m, 2H), 6.88 (s, 1H), 4.58 (s, 2H), 3.40 (q, 2H, J ~ 7 Hz), 2.70 (t, 2H, J ~ 7 Hz); 13 C NMR (150 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , 67.08, 38.46, 26.59; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 18 N 3 O , found N-(2-(3-cyano-4,6-dimethylpyridin-2-ylthio)ethyl)-2-(o-tolyloxy)acetamide (12): Compound 10jA (0.30 g, 1.43 mmol) was dissolved in absolute DCM (10 ml). Dry Et 3 N (0.20 ml, 1.43 mmol) was added, and the solution cooled to 0 o C. Neat mesylchloride (0.11 ml, 1.43 mmol) was combined portionwise within 10 min Stirring was continued for 2 h at the same temperature. A further share of dry Et 3 N (0.20 ml, 1.43 mmol) was added together with solid 4,6-dimethyl-2- thioxo-1,2-dihydropyridine-3-carbonitrile (thiopyridone 11, 0.24 g, 1.43 mmol), and the mixture 18

19 was stirred for 40 h at room temperature. DCM was evaporated under reduced pressure. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (once), diluted NaHCO 3 solution (2 times) and brine; dried over Na 2 SO 4 ; and then concentrated to dryness in vacuum. The crude product was chromatographed on silica gel (15 g) using hexane/ethyl acetate 2:1 as eluent. 146 mg (41% yield), of a white solid; 1 H NMR (500 MHz, CDCl 3 ) (m, 3H), 6.90 (dt, 1H, J = 0.8, 7.5 Hz), 6.72 (s, 1H), 6.71 (d, 1H, J = 8.2 Hz), 4.44 (s, 2H), 3.71 (q, 2H, J = 6.1 Hz), 3.47 (t, 2H, J = 6.1 Hz), 2.46 (s, 3H), 2.38 (s, 3H), 2.21 (s, 3H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , , 121,87, , , , , 67.90, 38.51, 29.64, 24.66, 20.08, 16.22; HRMS (ES+) m/z ([M + H] + ) calculated for C 19 H 22 N 3 O 2 S , found General Procedures for Preparation of Compounds 16E and 16F 2,5-dioxopyrrolidin-1-yl 2-chloroacetate (13): N-hydroxysuccinimide (1.15 g, 10 mmol) was dissolved in a mixture of absolute DCM (20 ml) and dry Et 3 N (1.39 ml, 10 mmol) at 0 o C. Neat 2-chloroacetyl chloride was combined portionwise within 5 min, whereupon a white precipitated was separated. The mixture was stirred for 1 h at 0 o C. The ice-bath was removed, and stirring was continued for 2 h. The reaction mixture was diluted with DCM, washed with ice-cold water (once), diluted NaHCO 3 solution (twice) and brine; dried over Na 2 SO 4 ; and concentrated in vacuum. The crude product showed excellent purity and no purification was carried out g (63%), pale solid; 1 H NMR (250 MHz, CDCl 3 ) 4.35 (s, 2H), 2.84 (s, 4H); 13 C NMR (60 MHz, CDCl 3 ) , , 37.79,

20 N-(2-(1H-indol-3-yl)ethyl)-2-(3-cyano-4,6-dimethylpyridin-2-ylthio)acetamide (16E): Tryptamine (54 mg, 0.34 mmol) was dissolved in ordinary DCM (30 ml) at 0 C. Solid 2,5- dioxopyrrolidin-1-yl 2-chloroacetate (13, 65 mg, 0.34 mmol) was added, and the mixture was stirred at 0 C. The reaction progress was monitored by TLC (hexane/ethyl acetate 2:3). After 2 h, tryptamine was still detected, but 13 was completely consumed. Additional 13 (30 mg) was added, and complete consumption of tryptamine was observed after an additional 30 min at 0 C. Ordinary Et 3 N (0.10 ml, 0.75 mmol) was added. After 30 min, excess 13 was no longer detected on TLC (hydrolysis in wet basic media). Solid 4,6-dimethyl-2-thioxo-1,2-dihydropyridine-3- carbonitrile (11, 55 mg, 0.34 mmol) was then added, and the mixture was stirred at room temperature overnight. 2-bromoacetic acid was added (to consume the minor amount of unreacted 11), and stirring was continued for 1 h. DCM was evaporated under reduced pressure. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (once), diluted NaHCO 3 solution (two times), and brine; dried over Na 2 SO 4 ; and concentrated to dryness in vacuum. The crude product was re-dissolved in DCM (ca. 1 ml), and then diethyl ether (5 ml) was added. The mixture was kept in a freezer overnight. The solid was filtered off, rinsed with diethyl ether, and dried in vacuum. 75 mg (60% yield), pale, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (s, 1H), 8.19 (t, 1H, J = 5.5 Hz), 7.50 (d, 1H, J = 7.7 Hz), 7.31 (d, 1H, J = 7.9 Hz), (m, 3H), 6.95 (t, 1H, J = 7.1 Hz), 3.93 (s, 2H), 3.60 (q, 2H, J ~ 6.6 Hz), 2.80 (t, 2H, J = 7.3 Hz), 2.38 (2s, 6H); 13 C NMR (100 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , , 39.86, 33.60, 25.04, 24.17, 19.61; HRMS (ES+) m/z ([M + H] + ) calculated for C 20 H 21 N 4 OS , found

21 N-(2-(1H-benzo[d]imidazol-2-yl)ethyl)-2-(3-cyano-4,6-dimethylpyridin-2-ylthio)acetamide (16F): Commercial 2-(2-aminoethyl)benzimidazole dihydrochloride (7, 80 mg, 0.34 mmol) was suspended in ordinary DCM (30 ml), and ordinary Et 3 N was added. After complete dissolution of 7, the mixture was cooled to 0 C, and solid 2,5-dioxopyrrolidin-1-yl 2-chloroacetate (13, 65 mg, 0.34 mmol) was added. The mixture was stirred at 0 C while the reaction progress was monitored by TLC (chloroform/methanol 20:3). Small amounts of 13 were added until complete consumption of 7 was observed. Solid 4,6-dimethyl-2-thioxo-1,2-dihydropyridine-3-carbonitrile (11, 55 mg, 0.34 mmol) was then added, and the mixture was stirred at room temperature overnight. 2-bromoacetic acid was added (to consume the minor amount of unreacted 11), and stirring was continued for 1 h. DCM was evaporated under reduced pressure. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (once), diluted NaHCO 3 solution (2 times), and brine; dried over Na 2 SO 4 ; and concentrated to dryness in vacuum. The crude product was suspended in diethyl ether, stirred at room temperature for 1 h, and kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 107 mg (86% yield), pale, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.27 (t, 1H, J ~ 5.4 Hz), (m, 2H), (m, 2H), 6.93 (s, 1H), 3.90 (s, 2H), 3.55 (q, 2H, J ~ 6.5 Hz), 2.92 (t, 2H, J ~ 6.9 Hz), 2.36 (s, 3H), 2.30 (s, 3H); 13 C NMR (100 MHz, DMSO-d 6 ) , , , , , , , , , , 37.31, 33.45, 28.56, 24.17, 19.57; HRMS (ES+) m/z ([M + H] + ) calcd for C 19 H 20 N 5 OS , found

22 Typical Procedures for Preparation of Compounds of Type 17 Preparation of N-hydroxysuccinimide esters. (2,5-dioxopyrrolidin-1-yl 2-(2- nitrophenoxy)acetate): 2-(o-nitrophenoxy)acetic acid (1 g, 15 mmol) was suspended in absolute DCM (30 ml) at room temperature. A catalytic amount of DMF (0.20 ml) was added, and then neat oxalyl chloride (2.28 g, 18 mmol) was combined portionwise within 10 min. Stirring was continued until gas evolution ceased. The clear solution that resulted was cooled to 0 C, solid N- hydroxysuccinimide (2.10 g, 18 mmol) was combined, and then neat dry Et 3 N (2.27 g, 22.5 mmol) was added portionwise within 10 min; stirring was continued for 1 h at 0 C. The reaction mixture was diluted with ordinary DCM (50 ml), acidified with a few drops of acetic acid, washed with ice-cold water (once), then washed with ice-cold diluted Na 2 CO 3 solution (3 times) and brine, dried over Na 2 SO 4 and concentrated to dryness in vacuum. The crude N- hydroxysuccinimide ester thus obtained was suspended in diethyl ether (10 ml), and DCM (1-2 ml) was added. The slurry was vigorously stirred at room temperature for 1 h and then kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum g (90% yield), pale solid; 1 H NMR (250 MHz, DMSO-d 6 ) 7.90 (m, 1H), 7.67 (m, 1H), 7.37 (m, 1H), 7.20 (m, 1H), 5.24 (s, 2H), 2.82 (s, 4H); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , 63.86, (2-(2-nitrophenoxy)acetamido)propanoic acid (17g): -alanine (0.67 g, 7.50 mmol), potassium carbonate (1.13 g, 8.16 mmol), tetrabutylammonium bromide (0.22 g, 0.68 mmol), ethylacetate (30 ml), and water (20 ml) were vigorously stirred. The solid N- hydroxysuccinimide ester described above (2,5-dioxopyrrolidin-1-yl 2-(2-nitrophenoxy)acetate) 22

23 was added portionwise under vigorous stirring within 30 min at room temperature. The slurry was stirred for one further hour, acidified with aqueous 6 M HCl solution, and extracted with ethyl acetate (3 times). The combined organic phases were washed with diluted aqueous HCl solution (3 times) and brine, dried over Na 2 SO 4, and concentrated to dryness in vacuum. The crude product was suspended and stirred in diethyl ether (10 ml) for 1 h. The resulted slurry was kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum g (87% yield), pale yellow crystals; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 7.97 (t, 1H, J = 5.3 Hz), 7.90 (dd, 1H, J = 8.0 Hz, J = 1.6 Hz), 7.63 (m, 1H), 7.22 (d, 1H, J = 8.5 Hz), 7.14 (m, 1H), 4.69 (s, 2H), 3.34 (q, 2H, J ~ 6 Hz), 2.41 (t, 2H, J = 6.8 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , 67.67, 34.55, (2-(o-tolyloxy)acetamido)propanoic acid (17j): 85% yield, white solid; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 7.92 (t, 1H, J = 5.1 Hz), (m, 2H), (m, 2H), 4.45 (s, 2H), 3.35 (q, 2H, J ~ 6.6 Hz), 2.42 (t, 2H, J = 6.6 Hz), 2.21 (s, 3H); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , 67.09, 34.44, 33.71, 16.07; HRMS (ES+) m/z ([M + H] + ) calculated for C 12 H 16 O 4 N , found (2-(1-bromonaphthalen-2-yloxy)acetamido)propanoic acid (17r): 2.80 g (80% yield), white, microcrystalline solid; 1 H NMR (250 MHz, CDCl 3 ) (bs, 1H), (m, 4H), 7.63 (m, 1H), (m, 2H), 4.73 (s, 2H), 3.39 (q, 2H, J = 6.3 Hz), 2.46 (t, 2H, J = 6.8 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , 68.35, 34.50,

24 3-(2-(naphthalen-2-yloxy)acetamido)propanoic acid (17n): 1 H NMR (500 MHz, DMSO-d 6 ) (bs, 1H), 8.21 (t, 1H, J = 5.8 Hz), 7.83 (m, 2H), 7.78 (dd, 1H, J = 8.1 Hz, J = 0.5 Hz), 7.45 (ddd, 1H, J = 8.2 Hz, J = 6.8 Hz, J = 1.2 Hz), 7.35 (ddd, 1H, J = 8.2 Hz, J = 6.8 Hz, J = 1.2 Hz), 7.28 (m, 1H), 7.24 (dd, 1H, J = 8.9 Hz, J = 2.5 Hz), 4.58 (s, 2H), 3.16 (dd, 2H, J = 6.9 Hz, J = 5.8 Hz), 2.45 (t, 2H, J = 6.9 Hz); 13 C NMR (125 MHz, DMSO-d 6 ) , , , , , , , , , , , , 67.00, 34.61, General Procedures for Preparation of Bisamides 18 The matching substituted propanoic acid of type 17 (1.0 equiv.) was dissolved in absolute DCM (50 ml) containing dry Et 3 N (1.0 equiv.) at 0 C under an Ar-atmosphere. Neat PivCl (1.0 equiv.) was added dropwise within 15 min, whereupon a white precipitate separated. The slurry was stirred for 20 min, and then the ice-bath was removed, and stirring was continued for 1 h. The reaction mixture containing the mixed anhydride was kept at 0 C. In a separate reaction flask, the matching aromatic amines (1.2 equiv. of G, H, or L and 2.4 equiv. of J or K) were dissolved in absolute THF under an Ar-atmosphere. Dry Et 3 N (1.0 equiv.) was added, and the solution was cooled to -78 C. The cold DCM solution containing the preformed mixed anhydride was carefully transferred to the reaction mixture by means of a syringe, and stirring at -78 C was continued for 2 h. The reaction mixture was concentrated under reduced pressure. The evaporation residue was portioned between ethyl acetate and water. The organic layer was washed with water (three times), diluted NaHCO 3 solution (two times) and brine; dried over Na 2 SO 4 ; and concentrated to dryness in vacuum to produce the crude products. 24

25 N-(2-aminophenyl)-3-(2-(2-nitrophenoxy)acetamido)propanamide (18gL): The crude product was suspended and stirred in ethyl acetate (2 ml) for 1 h. Diethyl ether (4 ml) was added and the slurry kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 365 mg (69% yield), brown, semi-crystalline solid; 1 H NMR (250 MHz, DMSO-d 6 ) 9.12 (s, 1H), 8.06 (t, 1H, J = 5.3 Hz), 7.90 (dd, 1H, J = 1.6, J = 8.1 Hz), 7.59 (m, 1H), 7.23 (d, 1H, J = 8.2 Hz), (m, 2H), 6.89 (m, 1H), 6.70 (dd, 1H, J = 1.1, J = 7.9 Hz), 6.43 (m, 1H), 4.83 (s, 2H), 4.71 (s, 2H), 3.44 (q, 2H, J ~ 6.4 Hz), 2.54 (t, 2H, J ~ 6.8 Hz); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , 67.69, 35.50, 35.13; HRMS (ES+) m/z ([M + H] + ) calculated for C 17 H 19 N 4 O , found N-benzyl-3-(2-(o-tolyloxy)acetamido)propanamide (18jG): The crude product was dissolved in DCM, and precipitated with diethyl ether. The solvent mixture was evaporated to dryness in vacuum, and the evaporation residue re-suspended and stirred in diethyl ether. The slurry was kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 270 mg (83% yield), snow-white, fluffy solid; 1 H NMR (250 MHz, CDCl 3 ) 7.39 (bs, 1H), (m, 5H), (m, 2H), 6.92 (t, 1H, J = 7.3 Hz), 6.71 (d, 1H, J = 8.1 Hz), 6.23 (bs, 1H), 4.40 (d, 2H, J = 5.2 Hz), 4.39 (s, 2H), 3.64 (q, 2H, J ~ 6.0 Hz), 2.47 (t, 2H, J ~ 6.0 Hz), 2.29 (s, 3H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , , , , , , 67.16, 43.55, 35.38, 34.99, 16.30; HRMS (ES+) m/z ([M + H] + ) calculated for C 19 H 23 O 3 N , found

26 N-phenyl-3-(2-(o-tolyloxy)acetamido)propanamide (18jH): The crude product was dissolved un BCM, and precipitated with diethyl ether. The solvent mixture was evaporated to dryness in vacuum and the evaporation residue re-suspended and stirred in diethyl ether. The slurry was kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 275 mg (88% yield), snow-white, amorphous solid; 1 H NMR (250 MHz, CDCl 3 ) 8.17 (bs, 1H), (m, 2H), 7.43 (bs, 1H), (m, 2H), (m, 3H), 6.38 (t, 1H, J = 7.3 Hz), 6.70 (d, 1H, J = 8.1 Hz), 4.46 (s, 2H), 3.72 (q, 2H, J ~ 5.9 Hz), 2,65 (t, 2H, J ~ 5.9 Hz), 2.26 (s, 3H); 13 C NMR (60 MHz, CDCl 3 ) , , , , , , , , , , , , 67.25, 36.57, 34.97, 16.26; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 21 O 3 N , found N-(4-hydroxyphenyl)-3-(2-(o-tolyloxy)acetamido)propanamide (18jJ): The crude product was suspended and stirred in ethyl acetate (2 ml) for 1 h. Diethyl ether (4 ml) was added and the slurry was kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 207 mg (63% yield), white, amorphous solid; 1 H NMR (250 MHz, DMSO-d 6 ) 9.69 (s, 1H), 9.13 (s, 1H), 7.98 (t, 1H, J = 5.5 Hz), 7.34 (d, 2H, J = 8.8 Hz), (m, 2H), (m, 2H), 6.67 (d, 2H, J = 8.8 Hz), 4.45 (s, 2H), 3.42 (q, 2H, J ~ 6.5 Hz), 2.47 (t, 2H, J ~ 6.5 Hz), 2.19 (s, 3H); 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , 67.12, 35.83, 34.98, 16.06; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 21 N 2 O , found N-(2-hydroxyphenyl)-3-(2-(o-tolyloxy)acetamido)propanamide (18jK): The crude product was suspended and stirred in diethyl ether (8 ml) for 1 h. The slurry was kept in a freezer 26

27 overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum g (75% yield), pale, amorphous solid; 1 H NMR (500 MHz, DMSO-d 6 ) 9.70 (s,1h), 9.28 (s, 1H), 7.98 (t, 1H, J = 5.4 Hz), 6.70 (d, 1H, J = 7.3 Hz), 7.13 (d, 1H, J = 7.3 Hz), 7.09 (t, 1H, J = 7.8 Hz), 6.93 (t, 1H, J = 7.1 Hz), (m, 3H), 6.75 (dt, 1H, J = 1.1, 7.6 Hz), 4.47 (s, 2H), 3.44 (q, 2H, J ~ 6.5 Hz), 2.62 (t, 2H, J ~ 6.5 Hz), 2.20 (s, 3H); 13 C NMR (60 MHz, DMSOd 6 ) , , , , , , , , , , , , , , 67.13, 35.71, 34.99, 16.06; HRMS (ES+) m/z ([M + H] + ) calculated for C 18 H 21 N 2 O , found N-(2-hydroxyphenyl)-3-(2-(naphthalen-2-yloxy)acetamido)propanamide (18nK): The crude product was suspended and stirred in diethyl ether (8 ml) for 1 h. The slurry was kept in a freezer overnight. The solid material was filtered off, rinsed with diethyl ether, and dried in vacuum. 431 mg (41% yield), pale yellow powder; 1 H NMR (250 MHz, DMSO-d 6 ) 9.70 (s, 1H), 9.29 (s, 1H), 8.23 (t, 1H, J ~ 5.1 Hz), (m, 4H), (m, 7H), 4.60 (s, 2H), 1.99 (q, 2H, J ~ 6.4 Hz), 2.64 (t, 2H, J ~ 6.7 Hz), 13 C NMR (60 MHz, DMSO-d 6 ) , , , , , , , , , , , , , , , , , , 67.05, 35.80, 35.12; HRMS (ES+) m/z ([M + H] + ) calculated for C 21 H 20 N 2 O 4 Na , found Typical Procedure for Preparation of Compounds 19M-O 2-(1-bromonaphthalen-2-yloxy)-N-(2-(5-chloro-1H-benzo[d]imidazol-2-yl)ethyl)acetamide (19N): The substituted propionic acid 17r (1. 0 g, 2.85 mmol) was suspended in absolute DCM 27

28 (30 ml) at room temperature. A catalytic amount of DMF (0.10 ml) was added, and then neat oxalyl chloride (0.54 g, 4.27 mmol) was combined portionwise within 10 min. Stirring was continued until gas evolution ceased (ca. 1 h). The clear solution that resulted was cooled to 0 C for addition of solid 4-chloro-2-nitroaniline (N, 0.49 g, 2.85 mmol). Neat, dry Py (0.69 ml, 8.55 mmol) was then combined portionwise within 10 min. Stirring was continued at 0 C for 30 min, acetic acid (2 ml) was added in one shot, and the DCM was evaporated in vacuum. The wet evaporation residue was suspended in n-butanol (20 ml). Iron powder (0.48 g, 8.85 mmol) was added, and the slurry was heated to reflux temperature for 3 h. The oil bath was removed, and after some time, ethyl acetate (150 ml) was carefully added to the still hot (ca C) reaction mixture under vigorous stirring. The resulting dark brown slurry was allowed to cool to room temperature, and the solid material was filtered out (suction through a paper filter). The limpid filtrate was washed with water (three times), diluted aq. sodium carbonate solution (three times), and brine. The organic phase was vigorously stirred in the presence of anhydrous Na 2 SO 4 for a few minutes, and then silica gel (ca. 1 g) was added, and stirring continued for a few minutes. The solid material was filtered off, and the solution was concentrated in vacuum. Finally, the bulk part of residual n-butanol was co-evaporated with ethanol. The wet crude product was suspended in a few milliliters of ethanol, and the slurry was kept in the freezer overnight. The precipitate was filtered off, rinsed with cold ethanol, and dried in vacuum. 735 mg (57% overall yield, based on acid 17r), pale, amorphous solid, excellent purity according to 1 H NMR; 1 H NMR (250 MHz, DMSO-d 6 ) (bs, 1H), 8.20 (t, 1H, J ~ 5 Hz), 8.05 (d, 1H, J = 8.5 Hz), 7.87 (m, 2H), (m, 4H), (m, 1H), (m, 1H), 4.72 (s, 2H), 3.68 (q, 2H, J ~ 6 Hz), 3.04 (t, 2H, J = 6.5 Hz); 13 C NMR (100 MHz, DMSO-d 6 ) , , , , , , , , , , , , , 28