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1 Supporting Information The Discovery of 3,3 -spiro[azetidine]-2-oxo-indoline Derivatives as Fusion Inhibitors for Treatment of RSV Infection Weihua Shi 1, Zhigan Jiang 1, Haiying He 1 *, Fubiao Xiao 1, Fusen Lin 1, Ya Sun 1, Lijuan Hou 1, Liang Shen 1, Lixia Han 1, Minggao Zeng 1, Kunmin Lai 1, Zhengxian Gu 1, Xinsheng Chen 1, Tao Zhao 2, Li Guo 2, Chun Yang 2, Jian Li 1, Shuhui Chen 1 1 WuXi AppTec (Shanghai) Co., Ltd, 288 FuTe Zhong Road, Shanghai , P. R. China 2 Buchang Pharmaceutical, 50 Gaoxin Road, Gaoxin District, Xi'an , Shanxi Province, P. R. China Contents: 1. Chemical synthesis 2. Hep-2 cell protection and cytotoxicity assays 3. In vivo PK study 4. In vivo Efficacy in the RSV Infection Mouse Model: 1

2 1. Chemical synthesis Supplementary Material Unless otherwise stated, all reagents and anhydrous solvents were of the highest grade available and were purchased from commercial sources and used without further purification. All NMR spectra were recorded on Bruker Advance III 400 MHz FT-NMR spectrometer with a 5 mm BBO (F) probe or a Varian 400 MHz spectrometer with a 1 H/ 19 F/ 31 P/ 13 C 5 mm PFG 4Nuc probe. 1H chemical shifts (δ) were reported in parts per million with MeOH-d 4, DMSO-d 6, or CDCl 3 as the reference standard. Signal multiplicities are represented by s (singlet), d (doublet), dd (doublet of doublets), t (triplet), q (quadruplet), br (broad), bs (broad singlet), and m (multiplet). Analytical LC-MS analyses were conducted using Shimadzu LC-20AB pumps and an SPD-M20 PDA detector set at 220 and 254 nm, and the MS detection was performed with a MS-2010EV Micromass Platform LC spectrometer in electrospray ionization mode. Analytical SFC-MS analyses were conducted using Mettler pumps and an Agilent G1315B detector set at nm, and the MS detection was performed with an Agilent 6110 detector following the detailed column and eluting conditions specified below. Preparative reversed-phase (RP) HPLC was performed using a Gilson 322 pump, a Gilson 156 UV detector set at 220 and 254 nm, and a Gilson GX-281 liquid handler, following the column Boston Green ODS 150*30*5u column using gradient mixtures of water/0.1% formic acid and water (10%)/acetonitrile (90%)/0.1% formic acid. Normal-phase silica gel preparative purification was performed using an automated Combi-flash companion from ISCO with pre-packed silica gel cartridges supplied by Santai Technologies Inc. and Agela Inc. SFC separation was performed using a Berger SFC Analytic system on a Column: Chiralpak AD mm I.D., 3um; Mobile phase: A: CO 2 B:ethanol (0.05% DEA); Gradient: from 5% to 40% of B in 4.5min and hold 40% for 2.5 min, then 5% of B for 1 min; Flow rate: 2.8mL/min; Column temperature:40 o C. Chromatographic purifications were performed silica gel ( mesh). Analytical thin-layer chromatography (TLC) was performed using SGF254 TLC plates ( mm) supplied by Combinol Reagent (Yantai) Co., Ltd. Preparative TLC was performed using SGF254 TLC plates ( mm) supplied by Yucheng Chemical (Shanghai) Co., Ltd. All of the compounds were established by a variety of LC/MS, and NMR analytical techniques, and purities were >95% for all final products. EXPERIMENTAL SECTION General Procedure 1 (Preparation of 6a-s): N-alkylation with 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol To a solution of N-alkylation nucleophilic reagent R-NH (1.0 eq) in DMF was added NaH (1.2 eq) 2

3 under N 2 at 0, the resulting mixture was stirred at 0~25 over 0.5 hr, then 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (1.2 eq) was added thereto. The reaction mixture was stirred at room temperature for 1~4 hrs. The reaction mixture was quenched with water at 0, extracted with EtOAc. The combined organic layers were washed with brine, dried over Na 2 SO 4, filtered and concentrated under reduced pressure to give a residue. The residue was subjected to prep-hplc for purification to give the desired product 5a-s. General Procedure 2 (Preparation of 6a-h): substituted benzo[d]imidazole cyclization To a solution of varied substituted benzene-1,2-diamine (1.0 eq) in DMF was added DIPEA (1.4 eq) and HATU (1.2 eq), stirred at room temperature for 15 min, followed by 2-(2'-oxospiro[cyclopropane-1,3'-indolin]-1'-yl)acetic acid (1.0 eq) was added, the mixture stirred at room temperature for 1~4 hrs. Water was added and extracted with EtOAc. The combined organic layer was washed with water and brine, dried with anhydrous Na 2 SO 4, filtered and concentrated to give a couple of amide isomers which was used directed for the next step. To the couple of amide isomers was added AcOH, and the resulting mixture was allowed to stir at 90 for 1 hr. The mixture was concentrated, and the residue was dissolved in EtOAc, and washed with aq.nahco 3. The combined organic layer was washed with water and brine, dried with anhydrous Na 2 SO 4, filtered and concentrated to give the various substituted benzimidazole intermediates. To the substituted benzimidazoles in MeCN was added 4-bromobutyl-1-ol (1.2 eq.), NaI (0.2 eq.), K 2 CO 3 (2.0 eq.) and stirred at 80 for 4~8 hrs till the monitored TLC indicated the starting material was consumed completely. The mixture was filtered and the filtrate was concentrated, purified by silica gel chromatography or SFC separation to give (6a-h). 4-((2-nitrophenyl)amino)butan-1-ol, 2. 3

4 HO H N NO 2 To a solution of 1-fluoro-2-nitrobenzene (6.73 g, mmol, 1.0 eq.) in 1,4-dioxane (70 ml) was added 4-aminobutan-1-ol (5.1 g, mmol, 1.2 eq.) and triethylamine (7.23 g, mmol, 1.5 eq.) sequentially. The resulting solution was stirred at 22 for 2 hrs. The mixture was concentrated in vacuo and washed with water for twice, extracted with EtOAc. The organic layer was concentrated in vacuo to give 3 as yellow oil (9.31g, 92.7%). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.14 (m, 1H), 8.06 (dd, J = 1.3 Hz, 8.6 Hz, 1H), (m, 1H), 7.06 (d, J = 8.6 Hz, 1H), (m, 1H), 4.46 (t, J = 5.2 Hz, 1H), (m, 4H), (m, 2H), (m, 2H). HRMS calcd for C 10 H 14 N 2 O 3 [M+H] + : , found: ((2-aminophenyl)amino)butan-1-ol, 3. HO H N NH 2 To a solution of 4-((2-nitrophenyl)amino)butan-1-ol 3 (22.3 g, 106 mmol, 1.0 eq.) in MeOH (250 ml) was added Pd/C (3.0 g, 5% w/w). The resulting solution was stirred at room temperature for 12 hrs under H 2 (1 atm). The mixture was filtered and the filtrates were concentrated in vacuo to give 4 as a dark oil (14.7g, 92.7%). 1 H NMR (400 MHz, CDCl 3 ) δ (m, 1H), (m, 3H), 3.67 (t, J = 5.9 Hz, 2H), (m, 2H), 3.13 (t, J = 6.4 Hz, 2H), (m, 4H). HRMS calcd for C 10 H 16 N 2 O [M+H] + : , found: (2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 4. N N Cl OH To a solution of 4-((2-aminophenyl)amino)butan-1-ol, 4 (4.2 g, 23.3 mmol, 1.0 eq.) in 4N HCl (40 ml) was added 2-chloroacetic acid (3.3 g, 35.0 mmol, 1.5 eq.). The resulting solution was refluxed at 80 for 2 hrs. The mixture was adjusted ph to 6~7 using saturated aqueous Na 2 CO 3 4

5 and extracted with EtOAc(100 ml 3). The organic layer was dried over Na 2 SO 4 and concentrated in vacuo. The organic layer was purified by silica gel chromatography (PE:EtOAc=2:1) to give 5 as a light yellow solid (3.1 g, 55.7%). 1 H NMR (400 MHz, CDCl 3 ) δ 7.76 (d, J = 7.3 Hz, 1H), (m, 1H), (m, 2H), 4.86 (s, 2H), (m, 2H), 3.72 (t, J = 6.1 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 12 H 15 ClN 2 O [M+H] + : , found: '-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-indolin]-2'- one, 5a. According to general procedure 1, spiro[cyclopropane-1,3'-indolin]-2'-one (100 mg, 1.0 eq ) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (180 mg, 1.2 eq) were reacted together in DMF (2 ml). After workup and purification, the title product 5a was obtained (35 mg, 15.4%) as a white solid. 1 H NMR(400 MHz, Methanol-d 4 ) δ 7.62 (d, J = 7.6 Hz, 1H),7.50 (d, J = 7.6 Hz, 1H), (m, 2H), (m, 2H), (m, 2H), 5.34 (s, 2H), 4.32 (t, J = 7.6 Hz, 2H), 3.50 (t, J = 6.4 Hz, 2H), (m, 6H), (m, 2H). HRMS calcd for C 22 H 23 N 3 O 2 [M+H] + : , found: '-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-pyrrolo[2,3 -c]pyridin]-2'(1'h)-one, 5b. According to general procedure 1, spiro[cyclopropane-1,3'-pyrrolo[2,3-c]pyridin]-2'(1'h)-one (70 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (114 mg, 1.1 eq.) were reacted together in DMF (1.5 ml). After workup and purification, the title product was obtained 5

6 5b (40 mg, 25.3%) as a light yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.34 (s, 1H), 8.24 (d, J = 4.8 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.55 (d, J = 8.0 Hz, 1H), (m, 1H), (m, 1H), 7.15 (d, J = 5.0 Hz, 1H), 5.42 (s, 2H), 4.39 (t, J = 7.7 Hz, 2H), 3.56 (t, J = 6.3 Hz, 2H), (m, 4H), (m, 2H), (m, 2H). HRMS calcd for C 21 H 22 N 4 O 2 [M+H] + : , found: '-fluoro-1'-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-in dolin]-2'-one, 5c. According to general procedure 1, 6'-fluorospiro[cyclopropane-1,3'-indolin]-2'-one (50 mg, 1.0 eq) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (73 mg, 1.1 eq) were reacted together in DMF (1.5 ml). After workup and purification, the title product 5c was obtained (40 mg, 25.3%) as light yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.69 (d, J = 7.6 Hz, 1H),7.55 (d, J = 7.6 Hz, 1H), (m, 2H), (m, 2H),6.77 (t, J = 8.0 Hz, 1H), 5.37 (s, 2H), 4.37 (t, J = 8.0 Hz, 2H),3.55 (t, J = 6.4 Hz, 2H), (m, 6H), (m, 2H). HRMS calcd for C 22 H 22 FN 3 O 2 [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-3,3-dimethylindolin-2-one, 5d. According to general procedure 1, 3,3-dimethylindolin-2-one (111.0 mg, 1.1 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (150.0 mg, 1.0 eq.) were reacted together in DMF (3 ml). 8.8%) as light yellow solid. After workup and purification, the title product was obtained 5d (20 mg, 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.63 (d, J = 8.0 Hz, 1H), 7.53 (d, J = 8.0 Hz, 1H), (m, 6H), 5.27 (s, 2H), 4.37 (t, J = 8.0 Hz, 2H),3.57 (t, J = 8.0 Hz, 6

7 2H), (m, 2H), (m, 2H), 1.43 (s, 6H). HRMS calcd for C 22 H 25 N 3 O 2 [M+H] + : , found: (2-((3-cyclopropyl-1H-indol-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5e. According to general procedure 1, 1,4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.0 eq.) and 1H-indole-3-carbonitrile (71 mg, 1.2 eq.) were reacted together in DMF (2 ml). After workup and purification, the title product was obtained 5e (72 mg, 49.9%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.13 (s, 1H), (m, 3H), 7.53 (d, J = 7.8 Hz, 1H), (m, 4H), 5.87 (s, 2H), 4.28 (t, J = 7.4 Hz, 2H), 3.43 (t, J = 5.9 Hz, 2H), (m, 4H). HRMS calcd for C 21 H 20 N 4 O [M+H] + : , found: (2-((3-methyl-1H-indazol-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5f. According to general procedure 1, 3-methyl-1H-indazole (110 mg) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (200 mg) were reacted together in DMF (2 ml). After workup and purification, the title product 5f was obtained (40 mg, 25.3%) as light yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.73 (d, J = 8.4 Hz, 2H), 7.66 (d, J = 8.0 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H),7.49 (d, J = 8.0 Hz, 1H), 7.39 (t, J = 8.0 Hz, 1H), (m, 1H), 7.16 (t, J = 8.0 Hz, 1H), 5.90 (s, 2H), 4.27 (t, J = 7.6 Hz, 2H), 3.40 (t, J = 5.6 Hz, 2H), 2.57 (s, 3H), 1.38 (t, J = 3.6 Hz, 4H). HRMS calcd for C 20 H 22 N 4 O [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)benzo[d]oxazol-2(3H)-one, 5g. 7

8 According to general procedure 1, benzo[d]oxazol-2(3h)-one (68 mg, 1.2 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.0 eq.) were reacted together in DMF (2.0 ml). After workup and purification, the title product 5g was obtained (23.2 mg, 16.5%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.62 (d, J = 8.0 Hz, 1H), 7.55 (d, J = 8.0 Hz, 1H), (m, 3H), (m, 3H), 5.42 (s, 2H), 4.41 (t, J = 7.6 Hz, 2H), 3.56 (t, J = 6.4 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 19 H 19 N 3 O 3 [M+H] + : , found: (2-((1H-pyrrolo[2,3-c]pyridin-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5h. Step 1. According to general procedure 1, 5-bromo-1H-pyrrolo[2,3-c]pyridine (100.0 mg, 1.1 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (111 mg, 1.0 eq.) were reacted together in DMF (2 ml). After workup, the residue obtained was purified by prep-tlc (Developer: EtOAc) to afford the product 4-(2-((5-bromo-1H-pyrrolo[2,3-c]pyridin-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (150mg, 80.8%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.62 (s, 1H), 7.81 (s, 1H), (m, 2H), 7.55 (d, J = 8.0 Hz, 1H), (m, 2H), 6.66 (d, J = 2.8 Hz, 1H), 5.89 (s, 2H), 4.28 (t, J = 7.6 Hz, 2H),3.45 (t, J = 6.0 Hz, 2H), (m, 4H). HRMS calcd for C 19 H 19 BrN 4 O [M+H] + : , found: Step 2. To a solution of 4-(2-((5-bromo-1H-pyrrolo[2,3-c]pyridin-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 0.25 mmol) in MeOH (3.0 ml) was added Pd(OH) 2 /C (20 mg, 5% w/w) at 22. The resulting 8

9 mixture was stirred at 22 for 40 min. After the reaction was complete, the mixture was filtered and the filtrate was concentrated to give the crude product. The residue was purified by prep-hplc (0.05% formic acid in water and CAN from 20-55%) to give the title product 5h (22 mg, 27.4%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 9.02(s, 1H), 8.19 (d, J = 6.0 Hz, 1H), 7.94 (d, J = 2.8 Hz, 1H), 7.87 (d, J = 6.0 Hz, 1H),7.64 (d, J = 8.0 Hz, 1H),7.56 (d, J = 8.0 Hz, 1H), (m, 2H), 6.86 (d, J = 3.2 Hz, 1H), 6.00 (s, 2H), 4.33 (t, J = 7.6 Hz, 2H), 3.47 (t, J = 6.0 Hz, 2H), (m, 4H). HRMS calcd for C 19 H 20 N 4 O [M+H] + : , found: (2-((1H-pyrazolo[3,4-c]pyridin-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5i. Step 1. According to general procedure 1, 5-bromo-1H-pyrazolo[3,4-c]pyridine (75 mg, 0.91 eq) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.0 eq) were reacted together in DMF (2 ml). After workup, the obtained residue was purified by preparation TLC to afford a pair of isomers (90 mg, 16.5%) as yellow solid, which was used directed in the next step. HRMS calcd for C 18 H 18 BrN 5 O [M+H] + : , found: Step 2. To a solution of 4-(2-((5-bromo-1H-pyrazolo[3,4-c]pyridin-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (60 mg) in MeOH (2 ml) was added Pd(OH) 2 /C (10 mg, 5% w/w) under N 2. The suspension was degassed under vacuum and purged with H 2 several times, which was stirred at room temperature for 1 hr. The mixture was subjected to prep-hplc (0.05% formic acid in water and CAN from 20-55%) for purification to give the title product 5i (3.3 mg, 27.4%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 9.17 (s, 1H), 8.24 (d, J = 4.4 Hz, 2H), (m, 1H),7.80 (d, J = 1.2 Hz, 1H),7.63 (d, J = 7.6 Hz, 1H), 7.52 (d, J = 7.6 Hz, 1H), (m, 2H), 6.13 (s, 2H),4.38 (t, J = 8.0 Hz, 2H), 3.47 (t, J = 6.4 Hz, 2H), (m, 4H). HRMS calcd for C 18 H 19 N 5 O 9

10 [M+H] + : , found: (2-((1H-indazol-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5j. According to general procedure 1, 1H-indazole(59 mg, 1.2 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.0 eq.) were reacted together in DMF (2 ml). After workup and purification, the title product 5j was obtained (26 mg, 19%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.09 (s, 1H), 7.77(d, J = 8.4 Hz, 1H), 7.76(d, J = 5.6 Hz, 2H), 7.48(d, J = 8.4 Hz, 1H), 7.39 (t, J = 5.6 Hz, 1H), (m, 2H), 7.17 (t, J = 5.6 Hz, 1H), 5.97 (s, 2H), (m, 2H), (m, 2H), (m, 4H). HRMS calcd for C 19 H 20 N 4 O [M+H] + : , found: (2-((3-methyl-4,5,6,7-tetrahydro-2H-indazol-2-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1 -ol, 5k and 4-(2-((3-methyl-4,5,6,7-tetrahydro-2H-indazol-2-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1 -ol, 5l The flasks (50 ml) equipped with magnetic stirrer was charged with 1-(cyclohex-1-en-1-yl)ethanone (1.0 g, 8.06 mmol, 1.0 eq), hydrazine hydrate (2.02 g, 40.3 mmol, 5.0 eq) and EtOH (10 ml). The resulting solution was stirred at 80 for 18 hr. The solvent was removed directly under vacuum conditions. The residue was charged with dioxane (10 ml) and DDQ (2.01 g, 8.9 mmol, 1.1 eq), and the mixture was refluxed for 6 hrs. Then KOH (20% aq.) and DCM were added to the reaction mixture which stirred for 1 hr. The solvent was separated and the aqueous phase extracted with DCM (50.0 ml 3). The organic layer was 10

11 concentrated in vacuum and purified by silica gel chromatography (PE:EtOAc =1:1) to afford 3-methyl-4,5,6,7-tetrahydro-1H-indazole (371 mg, 33.73%) as yellow oil. According to general procedure 1, 3-methyl-4,5,6,7-tetrahydro-1H-indazole (100 mg, 1.0 eq) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (190.4 mg, 1.1 eq) were reacted together in DMF (2.0 ml). After workup and purification, the desired product 5k (17 mg, 7%) and 5l (14 mg, 6%) was obtained respectively as white solid. 5k, 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.63 (d, J = 8.0 Hz, 1H), 7.51 (d, J = 8.0 Hz, 1H), (m, 2H), 5.50 (s, 2H), (m, 2H), (m, 2H), 2.50 (t, J = 5.9 Hz, 2H), 2.38 (t, J = 5.8 Hz, 2H), 2.11 (s, 3H), (m, 4H), (m, 4H). HRMS calcd for C 20 H 26 N 4 O [M+H] + : , found: l, 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.62 (d, J = 7.8 Hz, 1H), 7.51 (d, J = 8.0 Hz, 1H), (m, 2H), 5.57 (s, 2H), (m, 2H), (m, 2H), 2.58 (t, J = 5.9 Hz, 2H), 2.41 (t, J = 5.9 Hz, 2H), 2.15 (s, 3H), (m, 4H), (m, 4H). HRMS calcd for C 20 H 26 N 4 O [M+H] + : , found: (2-((2-(trifluoromethyl)-1H-benzo[d]imidazol-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan- 1-ol, 5m. According to general procedure 1, 2-(trifluoromethyl)-1H-benzo[d]imidazole (94 mg, 1.2 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.0 eq.) were reacted together in DMF (2 ml). After workup and purification, the title product 5m was obtained (56 mg, 34%) as yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.88 (d,j = 6.0 Hz, 1H), (m, 5H), 7.32 (t, J = 8.0 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H), 6.06 (s, 2H), 4.40 (t, J = 8.0 Hz, 2H), 3.58 (t, J = 6.4 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 20 H 19 F 3 N 4 O [M+H] + : , found:

12 4-(2-((2-(thiazol-4-yl)-1H-benzo[d]imidazol-1-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol, 5n. According to general procedure 1, 4-(1H-benzo[d]imidazol-2-yl)thiazole (70.46 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (100 mg, 1.2 eq.) were reacted together in DMF (2 ml). After workup and purification, the title product 5n was obtained (30 mg, 17.69%) as white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 9.12 (d, J = 1.8 Hz, 1H), 8.47 (d, J = 1.8 Hz, 1H), 7.77 (d, J = 7.8 Hz, 1H), (m, 3H), (m, 3H), 7.2 (t, J = 8.0 Hz, 1H), 6.52 (s, 2H), 4.38 (t, J = 7.5 Hz, 2H), 3.53 (t, J = 6.3 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 22 H 21 N 5 OS [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-4-methylquinazolin-2(1H)-one, 5o. According to general procedure 1, 4-methylquinazolin-2(1H)-one (56 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (122 mg, 1.46 eq) were reacted together in DMF (2 ml). After workup and purification, the title product 5o was obtained (91 mg, 71.7%) as pale yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 8.16 (d, J = 8.0 Hz, 1H), 7.78 (t, J = 8.0 Hz, 1H), 7.56 (d, J = 8.0 Hz, 2H), 7.47 (d, J = 8.0 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), (m, 1H), (m, 1H), 5.85 (s, 2H), 4.47 (t, J = 8.0 Hz, 2H), 3.62 (t, J = 6.0 Hz, 2H), (m, 3H), (m, 2H), (m, 2H). HRMS calcd for C 21 H 22 N 4 O 2 [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-4-methylquinolin-2(1H)-one, 5p. 12

13 According to general procedure 1, 4-methylquinolin-2(1H)-one (100 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (165 mg, 1.1 eq.) were reacted together in DMF (2 ml). After workup and purification, the title product 5p was obtained (30 mg, 13.2%) as pale yellow solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.91 (d, J = 8.0 Hz, 1H), (m, 4H), (m, 3H), 6.69 (s, 1H), 5.91 (s, 2H), 4.43 (t, J = 7.6 Hz, 2H), 3.57 (t, J = 6.0 Hz, 2H),2.59 (s, 3H), (m, 2H), (m, 2H). HRMS calcd for C 22 H 23 N 3 O 2 [M+H] + : , found: (2-((3,4-dihydro-1H-pyrido[3,4-b]indol-9(2H)-yl)methyl)-1H-benzo[d]imidazol-1-yl)butan-1 -ol, 5q. Step 1. According to general procedure 1, tert-butyl 3,4-dihydro-1H-pyrido[3,4-b]indole-2(9H)-carboxylate (150 mg, 1.1 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (120 mg, 1.0 eq.) were reacted together in DMF (2 ml). After workup to afford the crude product tert-butyl 9-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-3,4-dihydro-1H-pyrido[3,4-b]indole-2( 9H)-carboxylate (240 mg) as yellow oil. HRMS calcd for C 28 H 34 N 4 O 3 [M+H] + : , found: Step 2. To a solution of the crude (120 mg, 1.0 eq.) in DCM (4 ml) was added TFA (2 ml) at 0. The resulting solution was stirred at room temperature for 1 hr. The mixture was concentrated in vacuo and the residue was subjected to prep-hplc (0.05% formic acid in water and CAN from 20-55%) for purification to give the title product 5q (43 mg, 97.7%) as a white solid. 1 H NMR 13

14 (400 MHz, Methanol-d 4 ) δ 7.62 (d, J = 8.0 Hz, 1H), (m, 3H), (m, 3H), 7.15 (t, J = 8.0 Hz, 1H), 5.74 (s, 2H), 4.23 (s, 2H), 4.17 (t, J = 8.0 Hz, 2H), 3.43 (t, J = 8.0 Hz, 2H), (m, 2H), (m, 2H), (m, 2H), (m, 2H). HRMS calcd for C 23 H 26 N 4 O [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-4-methyl-4,5-dihydro-1H-benzo[b][ 1,4]diazepin-2(3H)-one, 5r. Step 1. According to general procedure 1, tert-butyl 2-methyl-4-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepine-1-carboxylate (130 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (136 mg, 1.2 eq.) were reacted together in DMF (1.5 ml). After workup, the residue obtained was purified by prep-tlc to afford the product tert-butyl-5-((1-(4-hydroxybutyl)-1h-benzo[d]imidazol-2-yl)methyl)-2-methyl-4-oxo-2,3,4,5-tetra hydro-1h-benzo[b][1,4]diazepine-1-carboxylate (110 mg, 48.8%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl 3 ) δ 7.79 (d, J = 7.6 Hz, 1H), (m, 3H), (m, 1H), 6.75 (d, J = 8.0 Hz, 1H), 6.63 (d, J = 8.0 Hz, 1H), 6.36 (d, J = 7.6 Hz, 1H), 5.43 (d, J = 15.2 Hz, 1H), 4.59 (d, J = 15.2 Hz, 1H), (m, 1H), (m, 2H), (m, 1H), (m, 1H), (m, 1H), (m, 5H), 1.42 (s, 9H), 1.36 (t, J = 6.4 Hz, 3H). HRMS calcd for C 27 H 34 N 4 O 4 [M+H] + : , found: Step 2. To a solution of tert-butyl 5-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2-methyl-4-oxo-2,3,4,5-tetrahydro-1Hbenzo[b][1,4]diazepine-1-carboxylate(100 mg, 1.0 eq.) in CF 3 COOH/DCM (5 ml) was stirred at 20 for 2 hrs, The reaction was quenched with water, The aqueous phase was basified with aq.nahco 3 till ph = 8 and extracted with EtOAc (50 ml 2). The combined organic layer was 14

15 dried over Na 2 SO 4, concentrated to give crude product. The crude product was subjected to HPLC (0.05% formic acid in water and CAN from 20-55%) for purification to give the title product 5r (20 mg, 25%) as white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.64 (d, J = 7.2 Hz, 1H), 7.50 (d, J = 7.2 Hz, 1H), (m, 2H), 7.90 (d, J = 8.0 Hz, 1H), (m, 3H), (m, 2H), (m, 4H), (m, 1H), 2.81 (dd, J = 3.6, 14.0 Hz, 1H), (m, 1H), (m, 2H), (m, 2H), 1.30 (d, J = 6.0 Hz, 3H). HRMS calcd for C 22 H 26 N 4 O 2 [M+H] + : , found: ((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-1-phenyl-1,3,8-triazaspiro[4.5]deca n-4-one, 5s. Step 1. According to general procedure 1, tert-butyl tert-butyl 4-oxo-1-phenyl-1,3,8-triazaspiro[4.5]decane-8-carboxylate (80 mg, 1.0 eq.) and 4-(2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (75 mg, 1.2 eq.) were reacted together in DMF (1 ml). After workup to afford the crude product tert-butyl-3-((1-(4-hydroxybutyl)-1h-benzo[d]imidazol-2-yl)methyl)-4-oxo-1-phenyl-1,3,8-triazas piro[4.5]decane-8-carboxylate (110 mg) as yellow oil. 1 H NMR (400 MHz, CDCl 3 ) δ 7.75(d, J = 6.8 Hz, 1H), (m, 1H), (m, 2H), (m, 2H), 6.84 (t, J = 7.2 Hz, 1H), 6.72 (d, J = 8.0 Hz, 2H), 4.93 (s, 2H), 4.80 (s, 2H), 4.33 (t, J = 8.0 Hz, 2H), 3.73 (t, J = 6.0 Hz, 2H), 1.94 (t, J = 7.6 Hz, 2H), (m, 7H), 1.50 (s, 12H). HRMS calcd for C 30 H 39 N 5 O 4 [M+H] + : , found: Step 2. To a solution of tert-butyl tert-butyl 3-((1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-4-oxo-1-phenyl-1,3,8-triazaspiro[4.5]de cane-8-carboxylate (100 mg, 1.0 eq.) in TFA/DCM (5 ml) was stirred at 20 for 2 hrs. After the reaction was complete, the mixture was concentrated to remove TFA and DCM, The residue 15

16 was diluted with THF (2 ml) and added LiOH (23 mg, 0.94 mmol, 5 eq.), The mixture was stirred at 20 for 3 hrs. The reaction was quenched with water. The mixture was extracted with EtOAc (50 ml 2). The combined organic layer was dried over Na 2 SO 4, concentrated to give crude product. The crude product was subjected to prep-hplc (0.05% formic acid in water and CAN from 20-55%) to give the title product 5s (20 mg, 24.5%) as a white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.65(d, J = 8.0 Hz, 1H),7.58 (d, J = 8.0 Hz, 1H), (m, 4H), (m, 3H), 4.99 (s, 2H), 4.87 (s, 2H), 4.41 (t, J = 7.6 Hz, 2H), (m, 2H), 3.61 (t, J = 6.4 Hz, 2H), 3.40 (d, J = 9.6 Hz, 2H), (m, 2H),2.14 (d, J = 14.8 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 25 H 31 N 5 O 2 [M+H] + : , found: '-((5-chloro-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-in dolin]-2'-one, 6a. According to general procedure 2, to afford the product 6a as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.68 (d, J = 1.8 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.27 (dd, J = 1.9, 8.7 Hz, 1H), (m, 2H), (m, 2H), 5.29 (s, 2H), 4.48 (s, 1H), 4.31 (t, J = 7.7Hz, 2H), (m, 2H), (m, 6H), (m, 2H). HRMS calcd for C 22 H 22 ClN 3 O 2 [M+H] + : , found: '-((1-(4-hydroxybutyl)-5-methyl-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-in dolin]-2'-one, 6b. According to general procedure 2, to afford the product 6b as a white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.86 (d, J = 9.28 Hz, 1H), (m, 2H), (m, 1H), (m, 16

17 3H), 5.67 (s, 2H), (m, 2H), 3.62 (t, J = 6.16 Hz, 2H), 2.68 (s, 1H), 2.56 (s, 3H), 1.99 (td, J = 7.84, Hz, 2H), (m, 4H), (m, 2H). HRMS calcd for C 23 H 25 N 3 O 2 [M+H] + : , found: '-((5-cyclohexyl-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1, 3'-indolin]-2'-one, 6c. To the solution of 6h (300 mg, 0.68 mmol, 1.0 eq.), cyclohex-1-en-1-ylboronic acid (103 mg, 0.82 mmol, 1.2 eq.), Pd(PPh 3 ) 4 (61 mg, 0.07 mmol, 0.1 eq.) and Na 2 CO 3 (108 mg, 1.02 mmol, 1.5 eq.) in DMF (1 ml) was stirred under microwave at 90 under N 2 protection over 4 hrs. The reaction was quenched with water, extracted with EA. The combined organic layers were washed with water, dried over Na 2 SO 4, concentrated in vacuum. The residue was purified via column chromatography (PE/EA=1:1) to afford 150 mg of light yellow oil. The oil was dissolved in MeOH (4 ml) and Pd/C (30 mg, 5% w/w) was added. The mixture was stirred under H 2 balloon at room temperature overnight. The reaction mixture was filtered and the filtrate was concentrated in vacuum. The residue was purified by prep-hplc (0.05% formic acid in water and CAN from 20-65%) purification to affordedthe desired compound 6c (40 mg, 13% yield) as a white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ (m, 1H), (m, 4H), 7.32 (s, 1H), 7.21 (d,j = 8.5 Hz, 1H), 6.03 (s, 2H), 4.36 (t, J = 7.5 Hz, 2H), 3.56 (t, J = 6.3 Hz, 2H), (m, 1H), (m, 7H), (m, 11H). HRMS calcd for C 28 H 33 N 3 O 2 [M+H] + : , found: '-((5-fluoro-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-in dolin]-2'-one, 6d. 17

18 According to general procedure 2, to afford the product 6d as a white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ (m, 1H), (m, 1H), (m, 5H), 5.37 (s, 2H), 4.36 (t, J = 7.6 Hz, 2H), 3.54 (t, J = 6.4 Hz, 2H), (m, 6H), (m, 2H). HRMS calcd for C 22 H 22 FN 3 O 2 [M+H] + : , found: (4-hydroxybutyl)-2-((2'-oxospiro[cyclopropane-1,3'-indolin]-1'-yl)methyl)-1H-benzo[d]imid azole-5-carbonitrile, 6e. According to general procedure 2, to afford the product 6e as a white solid. 1 H NMR (400 MHz, CDCl 3 ) δ 8.10 (s, 1H) (q, J = 12Hz, 1H) (d, J = 8Hz, 1H) (d, J = 8.4Hz, 1H) (t, J = 16Hz, 1H) (t,j = 16 Hz, 1H) (d, J = 7.2Hz, 1H) 5.32 (s, 2H) (t, J = 15.6Hz, 2H) (q, J = 15.6Hz, 2H) (m, J = 7.6Hz, 2H) (m, J = 15.6Hz, 2H) (m, J = 13.6 Hz, 2H) (m, J = 4Hz, 2H). HRMS calcd for C 23 H 22 N 4 O 2 [M+H] + : , found: '-((1-(4-hydroxybutyl)-5-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cycloprop ane-1,3'-indolin]-2'-one, 6f. According to general procedure 2, to afford the product 6f as a pale white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H), (m, 2H), (m, 2H), 5.37 (s, 2H), 4.39 (t, J = 7.7 Hz, 2H), 3.38 (t, J = 6.4 Hz, 2H), 18

19 (m, 6H), (m, 2H). HRMS calcd for C 23 H 22 F 3 N 3 O 2 [M+H] + : , found: '-((5-(aminomethyl)-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropan e-1,3'-indolin]-2'-one, 6g. According to general procedure 2, to afford the product 6g as a white solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.84 (s, 1H), 7.71 (s, 1H), (d, J = 8.4Hz, 1H), (d, J = 8.4Hz, 1H), (d, J = 3.2Hz, 2H), (d, J = 7.2Hz, 1H), (d, J = 4.8Hz, 2H), 5.26 (s, 2H), 4.78 (s, 1H), (m, J = 15.2Hz, 2H), 4.05 (s, 1H), (d, J = 3.2Hz, 2H), 1.65(s, 2H), (m, J = 28.8Hz, 4H), (m, J = 21.2Hz, 2H). HRMS calcd for C 23 H 26 N 4 O 2 [M+H] + : , found: '-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[cyclopropane-1,3'-in dolin]-2'-one, 6h. According to general procedure 2, to afford the product 6h as a white solid. 1 H NMR (400 MHz, Methanol-d 4 ) δ 7.77 (s, 1H), (m, 2H), (m, 2H), (m, 2H), 5.28 (s, 2H), (m, 2H), (m, 2H), (m, 6H), (m, 2H). HRMS calcd for C 22 H 22 BrN 3 O 2 [M+H] + : , found: ((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2',3',5',6'-tetrahydrospiro [indoline-3,4'-pyran]-2-one, 14a 19

20 Step 1 1-iodo-2-(2-iodoethoxy)ethane (14a-2) To a mixture of compound 14a-1 (10 g, 70 mmol, 1.0 eq.) and NaI (30 g, 0.2 mol, 2.8 eq.) was stirred in acetone (100 ml) at 55 o C for 18 hours. The mixture was washed with water and extracted with EtOAc (3 times). The organic layer was washed with water and NaHCO 3 (aq.).the organic layer was dried over by Na 2 SO 4. The organic layer was removed under reduce pressure to afford crude product 16 g (70.2% yield). The crude product which was used next step without further purification. 1 H NMR (400 MHz, CDCl 3 ) δ 3.24 (t, J=6.72 Hz, 4 H) (m, 1 H) (m, 5 H). Step 2 tert-butyl 2-oxoindoline-1-carboxylate (14a-4) To a mixture of compound 14a-3 (20 g, 0.15 mol, 1.0 eq.) and DMAP (3.4g, mol, 0.19 eq.) in MeCN (200 ml) at 26 o C. Then (Boc) 2 O (41.9 g, mol, 1.3 eq.) was added dropwise at 26 o C. The mixture was stirred at 26 o C for 16 hours. The solvent was removed and washed with water. The mixture was extracted with EtOAc. The organic layer was washed with water and brine. The organic layer was dried over and concentrated. The crude product was purified by silica gel chromatography(pe : Ethyl acetate = 70 : 1) to afford 12g (34.2% yield) of compound 14a-4. 1 H NMR (400 MHz, CDCl 3 ) δ 1.67 (s, 9 H) 3.68 (s, 2 H) 7.17 (d, J=7.52Hz, 1 H) 7.26 (s, 1 H) (m, 1 H) 7.81 (d, J=8.28 Hz, 1 H). Step 3 tert-butyl 2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-1-carboxylate (14a-5) To a mixture of compound 14a-4 (4.395 g, mmol, 1.0 eq.), compound 14a-2 (6.035 g, mmol, 1.0 eq.) and Cs 2 CO 3 ( g, mmol, 4.0 eq.) in DMF (50 ml) was stirred at -20 o C for 16 hous. The mixture was with water and extracted with EtOAc (3 times). The organic layer was washed with water and NaHCO 3 (aq).the organic layer was dried over and concentrated. The crude product was purified by silica gel chromatography (PE : Ethyl acetate=25:1) to afford 20

21 2.455g (43.8% yield) of compound 14a-5. 1 H NMR (400 MHz, CDCl 3 ) δ (m, 9 H) (m, 2 H) 1.93 (dd, J=10.03, 4.40 Hz, 2 H) 3.91 (dt, J=11.92, 4.20 Hz, 2 H) (m, 2 H) 7.20 (d, J=7.32 Hz, 1 H) (m, 2 H) 7.83 (d, J=8.08 Hz, 1 H). Step 4 2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (14a-6) To a solution of TFA (7 ml) in DCM (14 ml) was stirred at 26 o C. The solution was added dropwise into compound 14a-5 at 26 o C. The solution was stirred at 26 o C for 30 min. The solution was with NaHCO 3 (aq.) and extracted with EtOAc 3 times. The organic layer was washed with brine.the organic layer was dried over and concentrated. The crude product was purified by silica gel chromatography (PE : Ethyl acetate :DCM = 1 : 1 : 1) to afford 900 mg (56.3% yield) of compound 14a-6. 1 H NMR (400 MHz, CDCl 3 ) δ (m, 4 H) 3.93 (dt, J=11.58, 4.92 Hz, 2 H) 4.24 (ddd, J=11.85, 7.55, 4.40 Hz, 2 H) 6.90 (d, J=7.72 Hz, 1 H) (m, 1 H) 7.23 (t, J=7.94 Hz, 1 H) 7.36 (d, J=7.52 Hz, 1 H) 7.94 (br. s., 1 H). Step 5 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2',3',5',6'-tetrahydrospiro [indoline-3,4'-pyran]-2-one (14a) To a mixture of compound 14a-6 (300 mg, mmol, 1.0 eq.) in DMF (50 ml) was added NaH (79 mg, 1.9 mmol, 1.3 eq.) at 26 o C. The mixture was stirred at 26 o C for 20 min. Then, to this solution, compound 7 (470 mg, mmol, 1.0 eq.) was added. The mixture was stirred at 26 o C for 1.5 hours. The mixture was with NH 4 Cl (aq.) and extracted with EtOAc (3 times). The organic layer was washed with brine.the organic layer was dried over Na 2 SO 4. The solvent was removed under reduce pressure afford crude product. The crude product was purified by preparative HPLC (0.05% formic acid in water and CAN from 20-65%) to afford 280 mg (39.2% yield) of 14a. 1 H NMR (400 MHz, CDCl 3 ) δ 7.94 (d, J=1.00 Hz, 1 H), 7.52 (d, J=7.76 Hz, 1 H), 7.40 (d, J=7.52 Hz, 2 H), (m, 2H), (m, 1 H), 5.24 (s, 2 H), (m, 4 H), (m, 2H), 3.72 (t, J=5.92 Hz, 2 H), (m, 4H), (m, 4H). HRMS calcd for C 24 H 26 BrN 3 O 3 [M+H] + : , found:

22 1'-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-4,5-dihydro-2H-spiro[fur an-3,3'-indolin]-2'-one, 14b Step 1 N-(2-bromophenyl)tetrahydrofuran-3-carboxamide (14b-2) To a stirred solution of tetrahydrofuran-3-carboxylic acid (8. 02 g, 70 mmol ) in DCM (50 ml) was added oxalyl dichloride (10 ml). And the mixture was stirred at 25 C for 2 hours. The solvent was removed, then the residue was dissolved in DCM (50 ml). To this solution, 14b-1 (12 g, 70 mmol) and Et 3 N (20 g, 210 mmol) were added. The mixture was stirred at 25 C for 2 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in EtOAc, washed with water and brine, dried over Na 2 SO 4, and concentrated under reduced pressure. The crude was purified by column chromatography on silica gel (eluting with PE/EA = 15:1) to give 14 g of compound 14b-2 (74 % yield). Step 2 N-(2-bromophenyl)-N-(4-methoxybenzyl)tetrahydrofuran-3-carboxamide (14b-3) To a stirred solution of 14b-2 (5.4 g, 20 mmol) in DMF (40 ml) was added NaH (1.6 g, 40 mmol) and PMBCl (4.68 g, 30 mmol). The mixture was stirred at 25 C for 12 hours. The reaction was quenched with water, extracted with EtOAc, washed with water and brine, dried over Na 2 SO 4, and concentrated under reduced pressure. The crude was purified by column chromatography on silica gel (eluting with PE / Ethyl acetate = 4:1) to give 8.0 g of compound 14b-3 (100 % yield). Step 3 1'-(4-methoxybenzyl)-4,5-dihydro-2H-spiro[furan-3,3'-indolin]-2'-one (14b-4) To a solution of compound 14b-3 (2.5 g, 6.4 mmol) in dioxane (6 ml) were added tricyclohexylphosphine (700 mg, 2.5 mmol) and t-buona (1.75 g, 12.5 mmol). To this solution, Pd(OAc) 2 (275 mmol, 1.22 mmol) was added. And the mixture was subjected to microwave at 75 C for 40 mins. The mixture was filtered and extracted with EtOAc, concentrated and the residue was purified by column chromatography on silica gel (eluting with PE / Ethyl acetate = 10: 22

23 1) to give 1.2 g of compound 14b-4 (61 % yield). Step 4 4,5-dihydro-2H-spiro[furan-3,3'-indolin]-2'-one (14b-5) To a stirred solution of compound 14b-4 (2.0 g, 6.5 mmol) in CF 3 SO 3 H(10 ml) was added. The mixture was stirred at 25 C for 2 hours. The reaction was quenched with water, extracted with EtOAc, washed with water and brine, dried over Na 2 SO 4, and concentrated under reduced pressure. The crude was purified by column chromatography on silica gel (eluting with PE/EA = 5:1) to give 0.96 g of compound 14b-5 (80% yield). Step 5 4,5-dihydro-2H-spiro[furan-3,3'-indolin]-2'-one (14b) To a stirred solution of compound 14b-5 (378 mg, 2.0 mmol) in DMF (10 ml) were added 4-(5-bromo-2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (634 mg, 2.0 mmol) and NaH (120 mg, 3.0 mmol). The mixture was stirred at 25 C for one hour. Then the reaction was quenched with H 2 O, extracted with EtOAc, washed with water and brine, dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was dissolved in DMSO (5 ml). The crude was purified by Prep-HPLC (0.05% formic acid in water and CAN from 20-65%) to obtain the target product 14b (450 mg, 48% yield). 1 H NMR (CD 3 OD, 400MHz) δ (m, 2H), (d, J = 8.2 Hz, 1H), (d, J = 7.2 Hz, 1H), (m, 1H), (m, 1H), (d, J = 7.2 Hz, 1H), 5.62 (s, 2H), (m, 2H), (m, 1H), (m, 1H), (d, J = 8.8 Hz, 1H), (d, J = 8.8 Hz, 1H), (m, 2H), (m, 1H), (m, 1H), (m, 2H), (m, 2H). HRMS calcd for C 23 H 24 BrN 3 O 3 [M+H] + : , found: ((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[indoline-3,3'-thietan] -2-one, 14c Step 1 tert-butyl 3,3-bis(hydroxymethyl)-2-oxoindoline-1-carboxylate (14c-1) 23

24 An aqueous 37% HCHO solution (11.3 ml) was added to a mixture of 14a-4 (12 g, 1.0 eq) and Na 2 CO 3 (1.0 g, 0.2 eq) in dioxane (100 ml), after finished the mixture was stirred at room temperature for 2 h. The solvent was removed and the residue was purified by column chromatography (PE: Ethyl acetate = 3: 1) to obtain the target product (10 g, 66% yield) as white solid. 1 H NMR (CDCl 3, 400 MHz) δ: (m, 1H), (m, 2H), (m, 1H), (m, 1H), (m, 4H), 1.67 (s, 9H). Step 2 tert-butyl 3,3-bis(((methylsulfonyl)oxy)methyl)-2-oxoindoline-1-carboxylate (14c-2) To a solution of 14c-1 (12 g, 1.0 eq) in DCM (100 ml) in ice-water bath was added Et 3 N (37.5 ml, 2.0 eq) through a syringe followed by MsCl (11.7 g, 2.0 eq) also through a syringe, after finished the resulting mixture was stirred at 0 o C for 2.5 h. After being checked with TLC, the solvent was removed under reduced pressure and the residue was purified by column chromatography (PE: Ethyl acetate = 5: 1) to obtain the target product (7 g, 38% yield) as a colorless oil. 1 H NMR (CDCl 3, 400MHz) δ: (d, J = 8.0 Hz, 1H), (m, 2H), (m, 1H), (d, J = 10.0 Hz, 2H), (d, J = 10.0 Hz, 2H), 2.92 (s, 6H), 1.65 (s, 9H). Step 3 spiro[indoline-3,3'-thietan]-2-one (14c-3) A mixture of 14c-2 (8 g, 1.0 eq), Na 2 S (2.12 g 1.5eq) in DMF (50 ml) was stirred at 105 o C for 16 h. After cooling, the mixture was poured into ice-water and saturated NH 4 Cl aqueous solution, extracted with EA (100 ml 3), the combined organic layer was dried over Na 2 SO 4, concentrated, and purified by column chromatography (PE: Ethyl acetate = 4 : 1) to obtain the target product (1.4 g, 41% yield) as a white solid. 1 H NMR (CDCl 3, 400MHz) δ (d, J = 7.2 Hz, 1H), 7.81 (brs, 1H), (m, 1H), (m, 1H), (d, J = 7.6 Hz, 1H), (d, J = 9.6 Hz, 2H), (d, J = 9.6 Hz, 2H). Step 4 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[indoline-3,3'-thietan] -2-one (14c) To a mixture of 14c-3 (80 mg, 1.1 eq) in DMF (4 ml) was added NaH (11 mg, 1.2 eq) in ice-water 24

25 bath, after stirred at room temperature for 10 min, 4-(5-bromo-2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (120 mg, 1.0 eq) was added, after finished the resulting solution was stirred at room temperature for 16 h. The reaction solution was added cold water and extracted with EtOAc (30mL 2), the combined organic layer was dried over Na 2 SO 4, concentrated and the residue was further purified by Prep-HPLC (0.05% formic acid in water and CAN from 20-65%) to obtain the target product (85 mg, 47% yield). 1 H NMR (CD 3 OD, 400MHz) δ: (d, J = 7.2 Hz, 1H), (m, 2H), (d, J = 8.8 Hz, 1H), (m, 1H), (m, 1H), (d, J = 7.6 Hz, 1H), 5.53 (s, 2H), (t, J = 7.6 Hz, 2H), (d, J = 9.6 Hz, 2H), (t, J = 6.4 Hz, 2H), (d, J = 9.6 Hz, 2H), (m, 2H), (m, 2H). HRMS calcd for C 22 H 22 BrN 3 O 2 S [M+H] + : , found: ((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[indoline-3,3'-thietan] -2-one 1',1'-dioxide, 14d Step 1 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[indoline-3,3'-thietan] -2-one 1',1'-dioxide (14d) To a stirred ice-cooled solution of compound 14c (100 mg, 1.0 eq) in a mixture of water (0.8 ml), methanol (5.4 ml), and acetone (1.6 ml) was added Oxone (320 mg, 2.0 eq), after finished the reaction mixture was stirred at room temperature for overnight. The mixture was poured into saturated aqueous NH 4 Cl solution and extracted with EtOAc, the combined organic layer was dried over Na 2 SO 4, and concentrated under reduced pressure, then the residue was purified by Prep-HPLC (0.05% formic acid in water and CAN from 20-65%) to obtain the target product (30 mg, 25.7%). 1 H NMR (CD 3 OD) δ: (d, J = 7.6 Hz, 1H), (m, 2H), (m, 25

26 1H), 7.39 (m, 1H), (m, 1H), (d, J = 8.0 Hz, 1H), 5.51 (s, 2H), (m, 2H), (m, 2H), (m, 2H), (m, 2H), (m, 2H), (m, 2H). HRMS calcd for C 22 H 22 BrN 3 O 4 S [M+H] + : , found: isopropyl 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2-oxospiro[indoline-3,3'-p yrrolidine]-1'-carboxylate, 14e Step 1 methyl 2-(2-nitrophenyl)acetate (14e-2) The compound 14e-1 (60 g, 0.33 mol) was stirring in 300 ml of methanol. Sulfuric acid (1 ml) was added and the mixture heated to reflux. After 14 h, the mixture was cooled and evaporated under reduced pressure to give clear yellow oil. The oil was brought up in EtOAc and washed up with saturated NaHCO 3. The organic layers were dried and evaporated to afford the compound 14e-2 (64g, 99.07%), which was used directly. 1 H NMR (400MHz, CDCl 3 ) δ 8.11 (d, J=7.9 Hz, 1H), (m, 1H), (m, 1H), 7.35 (d, J=7.5 Hz, 1H), 4.02 (s, 2H), 3.71 (s, 3H). Step 2 methyl 2-(2-nitrophenyl)acrylate (14e-3) To a solution of the compound 14e-2 (52 g, 0.27 mol) in toluene (416 ml) was added HCHO (20.8 g, 0.63 mol), Bu 4 NI (3.95 g, mmol) and K 2 CO 3 (109.2 g, 0.8 mol) at room temperature. The resulting mixture was stirred for 12 h at 80 C. After cooling to room temperature, water (500 ml) was added and the aqueous phase was extracted with EtOAc(500 ml 2). The collected organic extracts were dried, filtered and concentrated under reduced pressure to give the crude product, which was purified by flash chromatography to afford the compound 3 (51.2g, 92.5%). 1 H NMR (400MHz, CDCl 3 ) δ 8.12 (d, J=7.9 Hz, 1H), 7.65 (dt, J=1.3, 7.5 Hz, 1H), (m, 1H),

27 (dd, J=1.1, 7.7 Hz, 1H), 6.54 (s, 1H), 5.88 (s, 1H), (m, 3H). Step 3 methyl 1-benzyl-3-(2-nitrophenyl)pyrrolidine-3-carboxylate (14e-4) To the solution of compound 14e-3 (6.2 g, mmol) and TFA (66 mg, mmol) in CHCl 3 (5 ml) was added a solution of N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (8.25 g, mmol) in CHCl 3 (25 ml) at 0 o C and the reaction solution was stirred at 70 o C for 12 h. After cooling to room temperature, saturated NaHCO 3 (100 ml) was added and the aqueous phase was extracted with DCM (50 ml 2). The collected organic extracts were dried, filtered and concentrated under reduced pressure to give the crude product, which was purified by flash chromatography to afford the compound 14e-4 (7.4g, 75%). 1 H NMR (400MHz, CDCl 3 ) δ 7.88 (dd, J=0.9, 7.9 Hz, 1H), 7.77 (d, J=7.5 Hz, 1H), (m, 1H), (m, 5H), (m, 1H), (m, 1H), (m, 4H), (m, 1H), (m, 2H), 2.82 (d, J=10.6 Hz, 1H), (m, 1H), (m,1h). Step 4 methyl 1-benzyl-3-(2-nitrophenyl)pyrrolidine-3-carboxylate (14e-5) To a solution of 14e-4 (4.9 g, 14.5 mmol) in MeOH (1 L) was added Pd/C (1.0 g) and the suspension was degassed under vacumm and purged H 2 three times. The mixture was stirred at 50 o C under the atmosphere of H 2 at 50 psi for 4 h. The mixture was filtrated and evaporated under the reduced pressure to afforded the compound 14e-5 (2.7 g, 99.2%), which was used directly. Step 5 tert-butyl 2-oxospiro[indoline-3,3'-pyrrolidine]-1'-carboxylate (14e-6) To a solution of 14e-5 (2.8 g, 14.8 mmol) and Et 3 N (4.67 ml) in DCM (20 ml) was added to Boc 2 O (2.9 g, 13.3mmol) dropwise at 0 o C. The mixture was stirred at the temperature. After 20 min, the mixture was washed up with water and the organic layers was evaporated under reduced pressure and separated by silica gel column chromatography using (PE:EA=20:1- PE:EA=3:1) to afforded the compound 14e-6 (1.4g, 36.8%). 1 H NMR (400MHz, CDCl 3 ) δ (m, 1H), (m, 1H), 6.99 (t, J=7.3 Hz, 1H), 6.84 (d, J=7.5 Hz, 1H), (m, 3H), (m, 1H), (m, 1H), 1.98 (s, 1H), (m, 9H). 27

28 Step 6 tert-butyl 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2-oxospiro[indoline-3,3'-p yrrolidine]-1'-carboxylate (14e-7) To a mixture of Compound 14e-6 (0.5 g, 1.74 mmol) and K 2 CO 3 (0.7 g,7.29 mmol) in CH 3 CN (20 ml) was added to Compound 4-(5-bromo-2-(chloromethyl)-1H-benzo[d]imidazol-1-yl)butan-1-ol (0.60 g, 1.91 mmol) and the mixture was refluxed for 10 h. The mixture was filter and the filtrated was evaporated under reduced pressure and separated by gel column chromatography using (PE : Ethyl acetate=20:1 PE : Ethyl acetate=1:1) to afforded the compound 14e-7 (0.38g, 38.5%) as a white solid. 1 H NMR (400MHz, CDCl 3 ) δ 7.85 (d, J=1.3 Hz, 1H), (m, 1H), 7.31 (dd, J=1.5, 8.6 Hz, 1H), (m, 1H), 7.15 (d, J=8.4 Hz, 1H), 7.10 (d, J=6.6 Hz, 1H), (m, 1H), 5.38 (d, J=15.4 Hz, 1H), (m, 1H), 4.93 (d, J=15.4 Hz, 1H), (m, 1H), (m, 1H), (m, 1H), (m, 2H), (m, 4H), 3.48 (br. s., 1H), (m, 1H), (m, 1H), 1.74 (d, J=8.8 Hz, 2H), (m, 3H), (m, 9H). Step 7 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)spiro[indoline-3,3'-pyrroli din]-2-one (14e-8) To a solution of 14e-7 (0.16 g, mmol) in CH 3 OH (3 ml) was added to HCl/CH 3 OH(1 mol/l, 120 ml) dropwise at room temperature. The mixture was stirred at room temperature for 5 h and evaporated under reduced pressure to afford 14e-8 (0.13g, 98.5%) as a white solid. 1 H NMR (400MHz, CD 3 OD) δ7.88 (d, J=11.0 Hz, 1H), (m, 1H), 7.57 (d, J=7.5 Hz, 1H), (m, 1H), (m, 1H), 7.14 (d, J=7.9 Hz, 1H), 5.56 (s, 2H), 4.62 (t, J=7.7 Hz, 2H), (m, 3H), (m, 3H), (m, 1H), (m, 1H), 2.04 (quin, J=7.6 Hz, 2H), (m, 2H). Step 8 isopropyl 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-2-oxospiro[indoline-3,3'-p 28

29 yrrolidine]-1'-carboxylate (14e) To a solution of 14e-8 (90 mg, mmol) and Et 3 N (96.9 mg, 0.96 mmol) in DCM (10 ml) was added to isopropyl carbonochloridate (23.4 mg, 0.192mmol) dropwise at -10 o C. After 10 mins, the mixture was washed up with saturated NH 4 Cl and extract with EtOAc and the organic layers was evaporated under reduced pressure and separated by TLC using (Ethyl acetate : CH 3 OH = 40 : 1) to afford compound 14e (56mg, 52.8%). 1 H NMR (400MHz, MeOH-d 4 ) δ 7.73 (s, 1H), (m, 1H), (m, 1H), 7.26 (dd, J=7.3, 19.2 Hz, 2H), (m, 2H), (m, 2H), (m, 1H), 4.37 (t, J=7.5 Hz, 2H), (m, 6H), (m, 1H), (m, 1H), 1.85 (br. s., 2H), (m, 2H), (m, 6H). HRMS calcd for C 27 H 31 BrN 4 O 4 [M+H] + : , found: ((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-1'-isobutyrylspiro[indolin e-3,3'-pyrrolidin]-2-one, 14f Step 1 1'-isobutyrylspiro[indoline-3,3'-pyrrolidin]-2-one (14f-1) To a solution of 14e-5 (0.55 g, mmol) and Et 3 N(0.887 g, mmol) in DCM(25 ml) was added to isobutyryl chloride (0.312 g, mmol) at 0 C. After 10 min, the mixture was washed up with saturated NH 4 Cl and extracted with EA, concentrated under reduced pressure to afford the compound 14f-1 (0.750 g, 99.3%) as white oil. 1 H NMR (400MHz, CDCl 3 ) δ (m, 1H), (m, 1H), 7.08 (dd, J=3.3, 7.3 Hz, 1H), (m, 1H), (m, 1H), (m, 4H), (m, 2H), (m, 1H), (m, 6H). Step 2 1-((5-bromo-1-(4-hydroxybutyl)-1H-benzo[d]imidazol-2-yl)methyl)-1'-isobutyrylspiro[indolin e-3,3'-pyrrolidin]-2-one, (14f) To a mixture of 14f-1 (0.264 g, mmol) and 4-(5-bromo-2-(chloromethyl)-1H-benzo 29