Design, synthesis, and biological evaluation of first-in-class dual acting

Design, synthesis, and biological evaluation of first-in-class dual acting histone deacetylases (HDACs) and phosphodiesterase 5 (PDE5) inhibitors for the treatment of Alzheimer s disease Obdulia Rabal, 1,5 Juan A. Sánchez-Arias, 1,5 Mar Cuadrado-Tejedor, 2,3,5 Irene de Miguel, 1 Marta Pérez-González, 2 Carolina García-Barroso, 2 Ana Ugarte, 1 Ander Estella-Hermoso de Mendoza, 1 Elena Sáez, 1 Maria Espelosin, 2 Susana Ursua, 2 Tan Haizhong, 4 Wu Wei, 4 Xu Musheng, 4 Ana Garcia-Osta, 2,* and Julen Oyarzabal. 1,* SUPPORTING INFORMATION Table of Contents S1. Protocol for preparative HPLC purification methods.....s2 S2. Protocol for SFC purification method of 7a and 7b... S2 S3. Synthesis of intermediates 1 to 20...... S3 S4. HPLC purities and HRMS of final compounds........ S18 S5. HPLC traces for final compounds....... S21 S6. Figure S1..S45 S7. Table S1 S46 S1

S1. Protocol for preparative HPLC purification methods The HPLC measurement was performed using Gilson 281 or SHIMADZU preparative HPLC, an autosampler and a UV detector. The fractions were detected by LC-MS. The MS detector was configured with an electrospray ionization source. The source temperature was maintained at 300-350 o C. Method 1 Reverse phase HPLC was carried out on luna C18 (100 x 30 mm; 4 µm). Solvent A: water with 0.075% trifluoroacetic acid; Solvent B: acetonitrile with 0.075% trifluoroacetic acid. Gradient: At room temperature, 20% of B to 40% of B within 6 minutes at 25 ml/min; then 40% B at 25 ml/min over 2 minutes. Method 2 Reversed phase HPLC was carried out on luna (100 x 30 mm; 5 µm). Solvent A: water; Solvent B: acetonitrile. Gradient: At room temperature, 45% of B to 75% of B within 9.5 minutes; then 75% B over 2 minutes, Flow rate: 20 ml/min. Method 3 Reversed phase HPLC was carried out on luna (100 x 30 mm; 5 µm). Solvent A: water with 0.075% trifluoroacetic acid; Solvent B: acetonitrile. Gradient: At room temperature, 25% of B to 55% of B within 11 minutes; then 55% B over 2 minutes. Flow rate: 25 ml/min. S2. Protocol for SFC purification method of 7a and 7b Preparative separation was performed using Thar SFC Pre-80 instrument with a Chiralcel OD-H column (5mm, 3.0 cm id x 25 cm L). Solvent A: CO 2 ; Solvent B: IPA (isopropylantipyrine). Mobile phase 65% of A and 35% of B at 70 g/min. Back pressure: 100 bar. S2

S3. Synthesis of intermediates 1 to 20 Synthesis of ethyl 2-piperazin-1-ylpyrimidine-5-carboxylate (Int.1) Pd(dppf)Cl 2 K 2 CO 3 CH 3 CN, 80 ºC SI-1 Et 3 N, CO EtOH, 100 ºC SI-2 HCl/CH 2 Cl 2 Int. 1 tert-butyl 4-(5-bromopyrimidin-2-yl)piperazine-1-carboxylate (SI-1) To a solution of 5-bromo-2-chloro-pyrimidine (9.75 g, 50 mmol) in CH 3 CN (100 ml) were added tert-butyl piperazine-1-carboxylate (9.25 g, 50 mmol) and K 2 CO 3 (13.8 g, 100 mmol) and the reaction mixture was stirred at 80 ºC overnight. Then the reaction mixture was concentrated under vacuum and extracted with EtOAc. The organic phase was washed with water, dried over anhydrous Na 2 SO 4, filtered and concentrated to give compound SI-1 (15 g 87%). ESI-MS m/z 343 [M+H] + calc. for C 13 H 19 BrN 4 O 2 ethyl 2-(4-tert-butoxycarbonylpiperazin-1-yl)pyrimidine-5-carboxylate (SI-2) To a solution of SI-1 (6.00 g, 17.49 mmol) in EtOH (100 ml) were added Et 3 N (1.77 g, 2.43 ml) and Pd(dppf)Cl 2 (5 g, 6.83 mmol) and the reaction mixture was stirred at 100 ºC at 1.5 MPa under CO atmosphere overnight. The reaction mixture was filtrated and the filtrate was concentrated to give the desired product SI-2 (5 g, 85%). ESI-MS m/z 337 [M+H] + calc. for C 16 H 24 N 4 O 4 ethyl 2-piperazin-1-ylpyrimidine-5-carboxylate (Int. 1) A solution of SI-2 (2.50 g, 7.44 mmol) in HCl/CH 2 Cl 2 (4.0 M, 30 ml) was stirred at room temperature overnight. Then, the solution was concentrated to give the desired Int. 1 (1.7 g, 97%). 1 H NMR (MeOD, 400 MHz): δ 8.88 (s, 2H), 4.35 (q, 2H), 4.15 (m, 4H), 3.30 (m, 4H), 1.38 (t, 3H). ESI-MS m/z 237 [M+H] + calc. for C 11 H 16 N 4 O 2 S3

Synthesis of ethyl (E)-3-[4-(piperazin-1-ylmethyl)phenyl]prop-2-enoate (Int. 2) THF SI-3 AcOH, MeOH SI-4 Boc 2 O, Et 3 N CH 2 Cl 2 TFA CH 2 Cl 2 SI-5 Int. 2 ethyl (E)-3-(4-formylphenyl)prop-2-enoate (SI-3) To a solution of terephthalaldehyde (15 g, 0.11 mol) in anhydrous THF (100 ml) was added ethyl (triphenylphosphoranylidene)acetate (19.5 g, 56 mmol) and the reaction mixture was stirred at room temperature overnight. Then, the reaction mixture was concentrated and purified by column chromatography to give the desired product SI-3 (5 g, 44%). 1 H NMR (CDCl 3, 400 MHz): δ 10.04 (s, 1H), 7.92-7.90 (d, J = 8 Hz, 2H), 7.70-7.68 (m, 3H), 6.62-6.58 (d, J = 16 Hz, 1H), 4.30 (q, 2H), 1.36 (t, 3H). ESI-MS m/z 205 [M+H] + calc. for C 12 H 12 O 3 ethyl (E)-3-[4-(piperazin-1-ylmethyl)phenyl]prop-2-enoate (SI-4) To a solution of compound SI-3 (0.204 g, 1 mmol) in MeOH (10 ml) was added AcOH (0.1 ml) and piperazine (0.164 g, 2 mmol) and the reaction mixture was stirred at room temperature overnight. Then, NaBH 3 CN (189 g, 3 mmol) was added into the reaction mixture and stirred at room temperature for 2 hours. Then the reaction mixture was concentrated to give the desired compound SI-4 (250 mg, 91%). ESI-MS m/z 275 [M+H] + calc. for C 16 H 22 N 2 O 2 tert-butyl 4-[[4-[(E)-3-ethoxy-3-oxo-prop-1-enyl]phenyl]methyl]piperazine-1-carboxylate (SI-5) To a solution of compound SI-4 (250 mg, 0.91 mmol) in CH 2 Cl 2 (12 ml) was added Et 3 N (152 mg, 1.50 mmol) and Boc 2 O (236 mg, 1.09 mmol). The reaction mixture was stirred at room temperature overnight, and then concentrated. The residue was purified by column S4

chromatography to give SI-5 (320 mg, 94%). ESI-MS m/z 375 [M+H] + calc. for C 21 H 30 N 2 O 4 ethyl (E)-3-[4-(piperazin-1-ylmethyl)phenyl]prop-2-enoate (Int. 2) To a solution of compound SI-5 (320 mg, 0.86 mmol) in CH 2 Cl 2 (10 ml) was added TFA (1 ml). The reaction mixture was stirred at room temperature for 5 hours, and then concentrated to give the desired compound Int. 2 (200 mg, 85%). ESI-MS m/z 275 [M+H] + calc. for C 16 H 22 N 2 O 2 Synthesis of methyl 3-(4-piperidyl)propanoate (Int.3) Pd/C, H 2 THF EtOH, 40 ºC SI-6 Int. 3 ethyl (E)-3-(4-pyridyl)prop-2-enoate (SI-6) To a solution of commercially available pyridine-4-carbaldehyde (5.16 g, 48.22 mmol) in anhydrous THF (50 ml) was added ethyl (triphenylphosphoranylidene)acetate (20.13 g, 57.82 mmol) and the reaction mixture was stirred at room temperature overnight. Then the reaction mixture was concentrated and purified by column chromatography to give the desired compound SI-6 (3 g, 36%). 1 H NMR (CDCl 3, 400 MHz): δ 8.67-8.65 (d, J = 6.4 Hz, 2H), 7.62-7.58 (d, J = 16 Hz, 1H), 7.38-7.36 (d, J = 6.4 Hz, 2H), 6.62-6.58 (d, J = 16 Hz, 1H), 4.32 (q, 2H), 1.35 (t, 3H). ESI-MS m/z 178 [M+H] + calc. for C 10 H 11 NO 2 methyl 3-(4-piperidyl)propanoate (Int.3) A mixture of SI-6 (3 g, 16.95 mmol) and Pd/C (0.5 g) in EtOH (30 ml) was stirred at 40 o C under H 2 atmosphere (40 Psi) overnight. The mixture was filtrated and washed with MeOH. Then, the filtrate was concentrated under vacuum to give the desired compound Int. 3 (3 g, 99 crude%). 1 H NMR (MeOD, 400 MHz): δ 3.67 (s, 3H), 3.04 (m, 2H), 2.65 (t, 2H), 2.35 (t, 2H), 1.75 (m, 2H), 1.65 (m, 2H), 4.32 (q, 2H), 1.41 (m, 1H), 1.25 (m, 2H). ESI-MS m/z 172 [M+H] + calc. for C 9 H 17 NO 2. (The methyl ester is formed when washing with MeOH). S5

Synthesis of ethyl 2-(4-amino-1-piperidyl)pyrimidine-5-carboxylate (Int. 4) Pd(dppf) 2 Cl 2 K 2 CO 3 CH 3 CN, 80 ºC SI-7 Et 3 N, CO EtOH, 100 ºC SI-8 HCl/CH 2 Cl 2 Int. 4 tert-butyl N-[1-(5-bromopyrimidin-2-yl)-4-piperidyl]carbamate (SI-7) To a solution of 5-bromo-2-chloro-pyrimidine (9.75 g, 50 mmol) in CH 3 CN (100 ml) were added tert-butyl piperidin-4-ylcarbamate (9.95 g, 50 mmol) and K 2 CO 3 (13.8 g, 100 mmol) and the reaction mixture was stirred 80 o C overnight. Then, the reaction mixture was concentrated and extracted with EtOAc. The organic layer was washed with water, dried over anhydrous Na 2 SO 4, filtered and concentrated under vacuum to give the crude product which was purified through column chromatography to give SI-7 (15 g, 84%). ESI-MS m/z 357 [M+H] + calc. for C 14 H 21 BrN 4 O 2 ethyl 2-[4-(tert-butoxycarbonylamino)-1-piperidyl]pyrimidine-5-carboxylate (SI-8) To a solution of SI-7 (5.00 g, 14.00 mmol) in EtOH (100 ml) were added Et 3 N (1.77 g, 2.43 ml) and Pd(dppf)Cl 2 (512 mg, 0.7 mmol) and the reaction mixture was stirred at 100 o C at 1.5 MPa under CO atmosphere overnight. The reaction mixture was filtrated and the filtrate was concentrated to give the crude product which was purified through column chromatography to give desired SI-8 (3 g, 61%). ESI-MS (M+1): 351.2 calc. for C 17 H 26 N 4 O 4 ethyl 2-(4-amino-1-piperidyl)pyrimidine-5-carboxylate (Int. 4) A solution of SI-8 (3 g, 8.57 mmol) in HCl/CH 2 Cl 2 (4.0 M, 30 ml) was stirred at room temperature overnight. Then, the solution was concentrated to give the desired Int. 4 (2 g, 93%). 1 H NMR (CDCl 3, 400 MHz): δ 8.88 (s, 2H), 4.88 (m, 2H), 4.35 (q, 2H), 3.51 (m, 1H), 3.25 (m, 2H), 2.15 (m, 2H), 1.65 (m, 2H), 1.38 (t, 3H). ESI-MS m/z 251 [M+H] + calc. for C 12 H 18 N 4 O 2 S6

Synthesis of ethyl (E)-3-(4-aminophenyl)prop-2-enoate (Int. 5) Fe THF AcOH SI-9 Int. 5 ethyl (E)-3-(4-nitrophenyl)prop-2-enoate (SI-9) To a solution of 4-nitrobenzaldehyde (10 g, 66.23 mmol) in anhydrous THF (100 ml) was added ethyl (triphenylphosphoranylidene)acetate (34 g, 99.35 mmol) and the reaction mixture was stirred at room temperature overnight. Then, the reaction mixture was concentrated and purified by column chromatography to give the desired compound SI-9 (10 g, 68%). 1 H NMR (CDCl 3, 400 MHz): δ 8.28-8.26 (d, J = 8.8 Hz, 2H), 7.77-7.65 (m, 3H), 6.60-6.56 (d, J = 16 Hz, 1H), 4.32 (q, 2H), 1.38 (t, 3H). ESI-MS (M+1): 222.2 calc. for C 11 H 11 NO 4 ethyl (E)-3-(4-aminophenyl)prop-2-enoate (Int. 5) To a solution of SI-9 (2.07 g, 10 mmol) in AcOH (30 ml) was added Fe (1112 mg, 20 mmol) and the reaction mixture was stirred at room temperature overnight. Then, the mixture was filtrated and the residue was concentrated to give the Int. 5 (1.5 g, 78%). 1 H NMR (CDCl 3, 400 MHz): δ 7.64-7.60 (d, J = 15.6 Hz, 1H), 7.38-7.36 (d, J = 8.4 Hz, 2H), 6.68-6.66 (d, J = 8.4 Hz, 2H), 6.27-6.24 (d, J = 15.6 Hz, 1H) 4.32 (q, 2H), 1.38 (t, 3H). ESI-MS (M+1): 192.1 calc. for C 11 H 13 NO 2 Synthesis of methyl 4-(4-oxo-1-piperidyl)benzoate (Int. 6) Cu(OAc) 2, Et 3 N CH 2 Cl 2 HCl/EtOAc 70 ºC SI-10 Int. 6 methyl 4-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)benzoate (SI-10) To a solution of compound 1,4-dioxa-8-azaspiro[4.5]decane (1.43 g, 10 mmol) in CH 2 Cl 2 (20 ml) were added (4-methoxycarbonylphenyl)boronic acid (3.6 g, 20 mmol), Cu(OAc) 2 (1.82 g, 10 mmol) and Et 3 N (2 g, 20 mmol) and the mixture was stirred overnight. Then, S7

the mixture was quenched with water and extracted with CH 2 Cl 2, the organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude product which was purified by column chromatography to give compound SI-10 (1.2 g, 44%) as a pale yellow solid. ESI-MS m/z 278 [M+H] + calc. for C 15 H 19 NO 4 methyl 4-(4-oxo-1-piperidyl)benzoate (Int. 6) A solution of compound SI-10 (0.8 g, 2.9 mmol) in HCl/EtOAc (2.0 M, 10 ml) was stirred at 70 o C overnight, then concentrated to give the crude product which was purified by column chromatography to obtained pure Int. 6 (0.3 g, 44%) as white solid. ESI-MS m/z 234.2 [M+H] + calc. for C 13 H 15 NO 3 Synthesis of methyl 2-azaspiro[5.5]undecane-9-carboxylate (Int. 7) HCl/EtOAc Int. 7 A solution of O2-tert-butyl O9-methyl 2-azaspiro[5.5]undecane-2,9-dicarboxylate (200 mg, 0.64 mmol) in HCl/EtOAc (1.0 N, 15 ml) was stirred at room temperature for 1 hour, then concentrated to give the desired Int. 7 (120 mg, 89%) as yellow solid. ESI-MS m/z 212.1 [M+H] + calc. for C 12 H 21 NO 2 Synthesis of ethyl 4-hydroxycyclohexanecarboxylate (Int. 8) NaBH 4 MeOH, 0 ºC then rt Int. 8 To a solution of ethyl 4-oxocyclohexanecarboxylate (5.0 g, 29 mmol) in MeOH (50 ml) was added NaBH 4 (2.24 g, 59 mmol) at 0 o C in portions, then the mixture was stirred at room temperature for 1 hour. Then, the reaction was quenched with water and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous S8

Na 2 SO 4, filtered and concentrated to give Int. 8 (4.95 g, 99%) as a pale yellow oil. ESI-MS m/z 173 [M+H] + calc. for C 9 H 16 O 3 Synthesis of tert-butyl 4-hydroxypiperidine-1-carboxylate (Int. 9) Boc 2 O, Et 3 N CH 2 Cl 2 Int.9 To a solution of piperidin-4-ol (10.1 g, 0.1 mol) in anhydrous CH 2 Cl 2 (100 ml) were added Et 3 N (20.2 g, 0.2 mol) and di-tert-butyl dicarbonate (21.6 g, 0.1 mmol) and the reaction mixture was stirred at room temperature for 3 hours. Then, the reaction mixture was washed with water and extracted with CH 2 Cl 2. The organic layer was dried over anhydrous Na 2 SO 4, filtered and concentrated under vacuum to give Int. 9 (20 g, 99%). ESI-MS m/z 202 [M+H] + calc. for C 10 H 19 NO 3 Synthesis of bromo-(2-ethoxy-2-oxo-ethyl)zinc (Int. 10) Zn, TMSCl THF, rt then 40 ºC Int. 10 Zn powder (5.2 g, 80 mmol) was put into a 250 ml of three-neck flask under N 2 protection, and then TMSCl (0.5 ml, 3.96 mmol) dissolved in dry THF (20 ml) was injected into the flask. The suspension mixture was stirred at room temperature for 20 minutes, then ethyl 2- bromoacetate (6.5 ml, 58.62 mmol) dissolved in dry THF (50 ml) was dropped into the flask for about 30 minutes at room temperature. After the addition was complete, the reaction mixture was stirred at 40 o C for another 30 minutes and then was used for the next step directly. Synthesis of tert-butyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)piperidine-1-carboxylate (Int. 11) CH 3 P(C 6 H 5 ) 3 Br nbuli, THF SI-11 9-BBN-H THF, reflux Int. 11 S9

tert-butyl 4-methylenepiperidine-1-carboxylate (SI-11) To a solution of methyl triphenyl phosphonium bromide (59.03 g, 0.166 mol) in THF (300 ml) was added n-buli (66 ml, 2.5 M in THF, 166 mmol) at -78 ºC and the mixture was stirred for 1 hour. Then, tert-butyl 4-oxopiperidine-1-carboxylate (30 g, 0.151 mol) was added dropwise in THF (30 ml) at -78 ºC. After addition, the mixture was stirred at room temperature overnight. Then, the reaction was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude compound which was purified by column chromatography to obtain pure compound SI-11 (18 g, 61%) as a pale yellow oil. ESI-MS m/z 198 [M+H] + calc. for C 11 H 19 NO 2. tert-butyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)piperidine-1-carboxylate (Int. 11) Compound SI-11 (6 g, 31 mmol) was treated with a solution of 9-BBN-H (0.5 M in THF, 62 ml) and the mixture was heated at reflux for 4 hours. The resulting Int. 11 was immediately used for the synthesis of compound 43d. Synthesis of tert-butyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)piperidine-1-carboxylate (Int. 12) 9-BBN-H n-buli THF, - 78 ºC to rt THF, reflux SI-12 Int. 12 tert-butyl 4-methylenepiperidine-1-carboxylate (SI-12) To a solution of methyltriphenylphosphonium bromide (59.03 g, 0.166 mol) in THF (300 ml) was added n-buli (66 ml, 2.5 M, 0.166 mol) at -78 o C and the mixture was stirred at - 78 o C for 1 hour. Then, tert-butyl 4-oxopiperidine-1-carboxylate (30 g, 0.151 mol) in THF (30 ml) was added dropwise at -78 o C and the mixture was stirred at room temperature overnight. Then, the reaction was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude compound which was purified by column S10

chromatography to give pure compound SI-12 (18 g, 61%) as a pale yellow oil. ESI-MS m/z 198 [M+H] + calc. for C 11 H 19 O 2 tert-butyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)piperidine-1-carboxylate (Int. 12) Compound SI-12 (6 g, 31 mmol) was treated with a solution of 9-BBN-H in THF (0.5 M, 62 ml) and the mixture was heated at reflux for 4 hours. The resulting Int. 11 was immediately used for the synthesis of compound 43g. Synthesis of ethyl 2-[4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclohexyl]acetate (Int. 13) Pd/C NaH MeOH, H 2 THF, 0 ºC to rt SI-13 SI-14 HCl/EtOAc 70 ºC 9-BBN-H n-buli THF, reflux SI-15 THF, -70 ºC to rt SI-16 Int. 13 ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate (SI-13) To a solution of ethyl 2-diethoxyphosphorylacetate (9.5 g, 42.3 mmol) in THF (20 ml) was added NaH (1.7 g 60% in mineral oil, 42.3 mmol) at 0 o C and the mixture was stirred at 0 o C for 1 hour. Then, a solution of 1,4-dioxaspiro[4.5]decan-8-one (6 g, 38.5 mmol) in THF (5 ml) was added at 0 o C and the solution was stirred at room temperature overnight. The mixture was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude product which was purified by the column chromatography to give compound SI-13 (6.6 g, 76%) as a white solid. 1 H NMR (CDCl 3, 400 MHz): δ 5.64 (s, 1H), 4.20-4.05 (m, 2H), 4.05-3.90 (m, 4H), 3.05-2.90 (m, 2H), 2.40-2.30 (m, 2H), 1.80-1.70 (m, 4H), 1.30-1.15 (m, 3H). ESI-MS m/z 227.2 [M+H] + calc. for C 12 H 18 O 4 S11

ethyl 2-(1,4-dioxaspiro[4.5]decan-8-yl)acetate (SI-14) To a solution of compound SI-13 (6 g, 26.5 mmol) in MeOH (40 ml) was added Pd/C (3 g). The solution was stirred at room temperature for 3 hours under H 2 atmosphere (1 atm). The solution was filtered and the filtrate was concentrated to give compound SI-14 (5 g, 83%) as a white solid. ESI-MS m/z 229.2 [M+H] + calc. for C 12 H 20 O 4 ethyl 2-(4-oxocyclohexyl)acetate (SI-15) A solution of compound SI-14 (5 g, 21.9 mmol) in HCl/EtOAc (2.0 M, 10 ml) was stirred at 70 o C overnight. Then, the solution was concentrated to give the crude product which was purified by column chromatography to obtain pure compound SI-15 (3 g, 74%) as white solid. 1 H NMR (CDCl 3, 400 MHz): δ 4.20-4.05 (m, 2H), 2.40-2.35 (m, 4H), 2.35-2.15 (m, 3H), 2.10-2.00 (m, 2H), 1.50-1.35 (m, 2H), 1.30-1.15 (m, 3H). ESI-MS m/z 185.2 [M+H] + calc. for C 10 H 16 O 3 ethyl 2-(4-methylenecyclohexyl)acetate (SI-16) To a solution of methyltriphenylphosphonium bromide (2.94 g, 8.2 mmol) in THF was added n-buli (3.4 ml, 2.5 M, 8.2 mmol) at -70 o C and the mixture was stirred at 0 o C for 2 hours. Then a solution of SI-15 (1 g, 5.43 mmol) in THF was added at -70 o C and the mixture was stirred at room temperature overnight. Then, the reaction was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude product which was purified by the column chromatography to give compound SI-16 (0.5 g, 50%) as a white solid. 1 H NMR (CDCl 3, 400 MHz): δ 7.48-7.45 (m, 2 H) 4.61 (s, 2H), 4.20-4.05 (m, 2H), 2.40-2.30 (m, 1H), 2.30-2.25 (m, 2H), 2.00-1.85 (m, 2H), 1.85-1.75 (m, 2H), 1.30-1.15 (m, 3H), 1.15-1.00 (m, 2H). ESI-MS m/z 183.2 [M+H] + calc. for C 11 H 18 O 2 ethyl 2-[4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclohexyl]acetate (Int. 13) Compound SI-16 (500 mg, 2.74 mmol) was treated with a solution of 9-BBN-H in THF (0.5 M, 10 ml) and the mixture was heated at reflux for 4 hours. The resulting Int. 13 was immediately used for the synthesis of compound 43h. S12

Synthesis of ethyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclohexanecarboxylate (Int. 14) 9-BBN-H n-buli THF, -70 ºC to rt THF, reflux SI-17 Int. 14 ethyl 4-methylenecyclohexanecarboxylate (SI-17) To a solution of methyltriphenylphosphonium bromide (5.3 g, 15 mmol) in THF was added n-buli (6 ml, 2.5 M, 15 mmol) at -70 o C and the mixture was stirred at 0 o C for 2 hours. Then, a solution of ethyl 4-oxocyclohexanecarboxylate (1.7 g, 10 mmol) in THF was added to the solution at -70 o C and the mixture was stirred at room temperature overnight. The reaction was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude product which was purified by the column chromatography to give compound SI-17 (1.02 g, 60%) as a white solid. ESI-MS m/z 169.2 [M+H] + calc. for C 10 H 16 O 2 ethyl 4-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclohexanecarboxylate (Int. 14) Compound SI-17 (500 mg, 3 mmol) was treated with a solution of 9-BBN-H in THF (0.5 M, 10 ml), and the mixture was heated at reflux for 4 hours. The resulting Int. 14 was immediately used for the synthesis of compound 43i. Synthesis of methyl 3-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclopentanecarboxylate (Int. 15) 9-BBN-H n-buli THF, -70 ºC to rt THF, reflux SI-18 Int. 15 Synthesis of methyl 3-methylenecyclopentanecarboxylate (SI-18) To a solution of methyltriphenylphosphonium bromide (10.7 g, 30 mmol) in THF was added n-buli (12 ml, 2.5 M, 30 mmol) at -70 o C. The mixture was stirred at 0 o C for 2 S13

hours. Then, a solution of methyl 3-oxocyclopentanecarboxylate (3 g, 21 mmol) in THF was added at -70 o C and the mixture was stirred at room temperature overnight. Then the mixture was quenched with aqueous NH 4 Cl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude product which was purified by column chromatography to give compound SI-18 (0.7 g, 24%) as a white solid. ESI-MS m/z 141.1 [M+H] + calc. for C 8 H 12 O 2 methyl 3-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclopentanecarboxylate (Int. 15) Compound SI-18 (700 mg, 5 mmol) was treated with a solution of 9-BBN-H in THF (0.5 M, 10 ml), and the mixture was heated at reflux for 4 hours. The resulting Int. 15 was immediately used for the synthesis of compound 43j. Synthesis of tert-butyl 3-oxocyclobutanecarboxylate (Int. 16) DCC, CH 2 Cl 2 Int. 16 To a solution of 3-oxocyclobutanecarboxylic acid (11.4 g, 100 mmol) in anhydrous CH 2 Cl 2 (100 ml) were added DCC (31 g, 150 mmol) and 2-methylpropan-2-ol (8.9 g, 120 mmol) and the mixture was stirred at room temperature overnight. Then, the mixture was quenched with water and extracted with CH 2 Cl 2. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated in vacuum. The residue was purified by column chromatography to give pure Int. 16 (15.1 g, 89%). 1 H NMR (CDCl 3 400 MHz): δ 3.29-3.27 (m, 2H), 3.23-3.21 (m, 2H), 3.16-3.08 (m, 1H), 1.44 (s, 9H). Synthesis of ethyl 3-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclobutanecarboxylate (Int. 17) KOH EtOH, 100 ºC SI-19 CH 3 CH 2 I K 2 CO 3 DMF 9-BBN-H THF, reflux SI-20 Int. 17 S14

3-methylenecyclobutanecarboxylic acid (SI-19) To a mixture of 3-methylenecyclobutanecarbonitrile (15 g, 161 mmol) in EtOH (500 ml) was added KOH (30.70 g, 540 mmol) in one portion under N 2 and the mixture was stirred at 100 C for 12 hours. Then, the mixture was cooled to 25 C and the residue was poured into water. The aqueous phase was extracted with EtOAc and the combined organic phase was washed with saturated brine, dried with anhydrous Na 2 SO 4, filtered and concentrated in vacuum to afford compound SI-19 (12.00 g, 67%) as yellow oil. ESI-MS m/z 113.3 [M+H] + calc. for C 6 H 8 O 2 ethyl 3-methylenecyclobutanecarboxylate (SI-20) To a solution of compound SI-19 (12 g, 107 mmol) in DMF (250 ml) was added K 2 CO 3 (29.6 g, 214 mmol), then CH 3 CH 2 I (20 g, 128 mmol) was added slowly. The reaction mixture was stirred at 25 C for 12 hours. Then, the mixture was quenched with water and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude compound which was purified by column chromatography to give pure compound SI-20 (9.5 g, 63%) as a pale yellow oil. 1 H NMR (CDCl 3 400MHz): δ 4.79 (s, 2H), 4.17-4.12 (m, 2H), 3.12-2.89 (m, 5H), 1.27-1.24 (m, 3H). ESI-MS m/z 141.1 [M+H] + calc. for C 8 H 12 O 2 ethyl 3-(9-borabicyclo[3.3.1]nonan-9-ylmethyl)cyclobutanecarboxylate (Int. 17) Compound SI-20 (3 g, 21.4 mmol) was treated with a solution of 9-BBN-H (0.5 M in THF, 40 ml) and the mixture was heated at reflux for 4 hours. The resulting Int. 17 was immediately used for the synthesis of compound 43p. S15

Synthesis of ethyl 3-[4-(bromomethyl)phenyl]propanoate (Int. 18) NaBH 4 POT, n-bu 3 N, Pd(OAc) 2 DMF, 90 ºC SI-21 THF/MeOH, 0 ºC NBS, PPh 3 CH 2 Cl 2, 0 ºC to rt SI-22 Int. 18 ethyl 3-(4-formylphenyl)propanoate (SI-21) To a solution of 4-bromobenzaldehyde (6.0 g, 32 mmol) in DMF (30 ml) were added 3,3- diethoxyprop-1-ene (12.65 g, 96 mmol), tri-o-tolylphosphine (1.2 g, 3.84 mmol), n-bu 3 N (11.84 g, 64 mmol) and Pd(OAc) 2 (215 mg, 0.96 mmol) and the reaction mixture was stirred at 90 o C for 1 hour. Then, the mixture was diluted with 2 N HCl and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude compound which was purified by column chromatography to give pure compound SI-21 (3.78 g, 57%) as a yellow oil. ESI-MS m/z 207 [M+H] + calc. for C 12 H 14 O 3 ethyl 3-[4-(hydroxymethyl)phenyl]propanoate (SI-22) To a solution of compound SI-21 (2.2 g, 10.68 mmol) in THF/MeOH (3:1, 40 ml) was added NaBH 4 (619 mg, 16.29 mmol) in portions at 0 o C and the reaction mixture was stirred at 0 o C for 1 hour. Then, the mixture was extracted with CH 2 Cl 2. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the desired compound SI-22 (1.93 g, 86%) as a pale yellow oil. ESI-MS m/z 209 [M+H] + calc. for C 12 H 16 O 3 ethyl 3-[4-(bromomethyl)phenyl]propanoate (Int. 18) To a solution of compound SI-22 (1.93 g, 9.28 mmol) in anhydrous CH 2 Cl 2 (30 ml) was added PPh 3 (0.97 g, 3.71 mmol) and the mixture was cooled to 0 o C. Then, NBS (1.96 g, 11.14 mmol) was added and the mixture was stirred at room temperature overnight. After LC-MS showed the starting material was consumed completely, the mixture was extracted S16

with CH 2 Cl 2 and the organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give the crude compound which was purified by column chromatography to give pure Int. 18 (680 mg, 27%) as a pale yellow oil. ESI-MS m/z 271 [M+H] + calc. for C 12 H 15 BrO 2 Synthesis of ethyl 2-[4-(trifluoromethylsulfonyloxy)cyclohex-3-en-1-yl]acetate (Int. 19) PhN(SO 2 CF 3 ) 2 LiHMDS THF, -78 ºC to rt Int. 19 To a solution of ethyl 2-(4-oxocyclohexyl)acetate (1.0 g, 5.43 mmol) in anhydrous THF (30 ml) was added LiHMDS (5.7 ml, 1.0 M in THF, 5.70 mmol) at -78 o C and the mixture was stirred at the same temperature for 1 hour. Then, PhN(SO 2 CF 3 ) 2 (2.04 g, 5.70 mmol) in THF (20 ml) was added under N 2 protection. After addition, the mixture was stirred at room temperature overnight. Then, the reaction was quenched with aqueous KHSO 4, extracted with methyl tert-butyl ether and the organic layer was washed with 1.0 M aqueous NaOH, aqueous NH 4 Cl and brine. The organic phase was dried over anhydrous Na 2 SO 4, filtered and concentrated to give Int. 19 (1.50 g, 87%) as a yellow oil. 1 H NMR (CDCl 3 400 MHz): δ 5.66 (s, 1H), 4.12-4.05 (m, 2H), 2.35-2.23 (m, 5H), 2.21-2.03 (m, 1H), 1.90-1.83 (m, 2H), 1.49-1.42 (m, 1H), 1.23-1.18 (m, 3H). ESI-MS m/z 317 [M+H] + calc. for C 11 H 15 F 3 O 5 S Synthesis of methyl 5-bromofuran-2-carboxylate (Int. 20) MeI, K 2 CO 3 DMF Int. 20 To a solution of 5-bromofuran-2-carboxylic acid (4 g, 21 mmol) in DMF (50 ml) was added K 2 CO 3 (5.8 g, 42 mmol) and then MeI (4.44 g, 31.5 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 5 hours. Then the mixture was quenched with water and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na 2 SO 4, filtered and concentrated to give Int. 20 (3.5 g, 81%) as a pale yellow oil. ESI-MS m/z 205 [M+H] + calc. for C 6 H 5 BrO 3 S17

S4. LC purities and HRMS of final compounds HPLC-analysis was performed using a Shimadzu LC-20AB or LC-20AD with a Luna- C18(2), 5µm, 2.0*50mm column at 40 ºC and UV detection at 215, 220 or 254 nm. Three different methods were used (Methods 1, 2 and 3). UHPLC analysis was performed using an Acquity UPLC system (Waters Corp., Milford, MA, USA) with thermostatized autosampler. Separation was carried out on an Acquity UPLC TM BEH C18 column (50 mm 2.1 mm, 1.7 mm) at 40 ºC and UV detection at 215, 220 or 254 nm. One method was used (Method 4). High Resolution Mass Spectrometry (HRMS) of final compounds High resolution mass spectrometry, m/z determination, was performed by a nanolc Ultra HPLC system (Eksigent, Sciex, Ontario, Canada) coupled to a TripleTOF 5600+ system (Sciex, Ontario, Canada). Separation was carried out on an Acclaim PepMap TM 100 column (100 µm 2 cm, 5µm, 100Å, nanoviper, ThermoScientific) at 25 ºC. One method was used (Method 4) Method 1 Solvent A: water with 0.056% trifluoroacetic acid; Solvent B: acetonitrile with 0.056% trifluoroacetic acid. Gradient: After 0.1 minutes at the initial condition of 90% A and 10% B, solvent B was increased to 80% over 4 minutes, maintained at 80% for 0.9 minutes, then a linear gradient to initial conditions was applied for 0.02 minutes and maintained for 0.58 minutes to re-equilibrate the column, giving a cycle time of 5.50 minutes. Flow rate was 0.8 ml/min from 0.01 to 4.90 minutes, increased to 1.2 ml/min in 0.03 minutes and maintained until the end of the run. Method 2 Solvent A: water with 0.056% trifluoroacetic acid; Solvent B: acetonitrile with 0.056% trifluoroacetic acid. Gradient: After 0.4 minutes at the initial condition of 100% A, solvent B was increased to 60% over 4 minutes, maintained at 60% for 0.8 minutes, then a linear gradient to initial conditions was applied for 0.02 minutes and maintained for 0.68 minutes S18

to re-equilibrate the column, giving a cycle time of 5.90 minutes. Flow rate was 0.8 ml/min from 0.01 to 5.21 minutes, increased to 1.2 ml/min in 0.02 minutes and maintained until the end of the run. Method 3 Solvent A: water with 0.056% trifluoroacetic acid; Solvent B: acetonitrile with 0.019% trifluoroacetic acid. Gradient: After 0.1 minutes at the initial condition of 90% A and 10% B, solvent B was increased to 80% over 4 minutes, maintained at 80% for 2 minutes, then a linear gradient to initial conditions was applied for 0.01 minutes and maintained for 0.68 minutes to re-equilibrate the column, giving a cycle time of 7.80 minutes. Flow rate was 3 ml/ min. Method 4 Solvent A: water with 0.1% formic acid; Solvent B: acetonitrile. Gradient: After 0.1 minutes at the initial condition of 90% A and 10% B solvent B was increased from 5 to 100% over 8 minutes, maintained at 100% for 2 minutes, then a linear gradient to initial conditions was applied for 0.5 minutes and maintained for 1.5 minutes to re-equilibrate the column (A: 95%, B: 5%), giving a cycle time of 12 minutes. Flow rate was 0.6 ml/min (Acquity UHPLC system) and 0.6µL/min (nanolc Ultra HPLC system) during the complete run. LC purities and HRMS of final compounds Compound Method Rt m/z by HRMS Purity 7 1 2.73 440.2293 99.46 7a 1 2.80 440.2293 96.51 7b 4 3.74 440.2293 98.18 13a 3 2.78 548.2280 99.22 13b 3 2.78 562.2441 95.87 13c 4 3.85 598.2200 100.00 13d 3 2.94 636.2603 95.46 13e 3 3.29 547.2337 96.71 13f 4 3.63 612.2349 99.07 13g 3 3.29 553.1867 95.91 S19

21a 1 1.88 415.2089 99.17 21b 1 2.03 469.2557 97.90 21c 1 2.15 546.2825 97.39 21d 1 2.14 548.2730 99.49 21e 1 2.31 562.2883 97.98 21f 1 2.13 562.2883 95.93 30a 1 2.01 469.2560 99.32 30b 1 1.81 483.2717 97.04 30c 4 2.13 509.2875 98.56 30d 1 2.82 523.3030 97.03 30e 1 3.03 588.3042 94.60 30f 1 2.66 470.2404 100.00 30g 1 2.95 464.1993 100.00 37 4 4.11 549.2569 96.03 42 4 2.84 372.1671 98.29 48a 1 2.32 386.1825 95.49 48b 1 2.44 400.1980 96.83 48c 1 1.88 548.2737 98.56 48d 4 4.54 547.2775 99.19 48e 1 2.62 546.2828 100.00 48f 1 2.84 545.2879 98.87 48g 1 2.35 497.2879 98.97 48h 1 3.30 482.2765 99.56 48i1 4 4.87 468.2608 99.40 48i2 4 4.25 468.2608 95.47 48j 4 4.29 454.2449 98.89 48k 2 2.71 426.2135 99.60 48l 1 2.00 469.2558 99.58 48m 1 1.85 441.2248 95.32 48n 1 2.64 426.2138 97.27 48o 1 2.44 412.1980 98.71 52a 4 4.37 490.2448 99.03 52b 4 4.34 488.2301 98.90 52c 4 4.12 468.2605 98.64 52d 4 3.59 449.1935 95.00 52e 4 3.71 449.1935 94.62 52f 4 3.82 438.1776 97.43 S20

LC traces for final compounds Compound 7 2.44 5594 16172 0.26 2.56 5334 16673 0.27 2.73 1580119 6105126 99.46 Compound 7a 2.44 5347 16599 0.13 2.80 3772718 12013602 96.51 3.33 141956 385218 3.09 4.56 6709 32489 0.26 S21

Compound 7b λ: 220 nm, 1.2 nm 3.74 1100774 29549.73 98.18 3.91 5887 247.18 0.82 4.14 4416 83.07 0.28 4.48 11248 218.61 0.73 Compound 13a 2.67 1419 3560 0.11 2.78 1267092 3310630 99.22 2.85 12688 18427 0.55 2.98 1751 4118 0.12 S22

Compound 13b 2.66 4898 17244 0.42 2.78 1505378 3899166 95.87 2.84 27072 49380 1.21 3.03 16216 47103 1.16 3.26 1520 4158 0.10 3.35 19736 50099 1.23 Compound 13c λ: 254 nm, 1.2 nm 3.85 454596 12275.72 100.00 S23

Compound 13d 2.30 1562 3640 0.08 2.87 7837 25466 0.58 2.94 1343160 4180218 95.46 3.09 25492 131497 3.00 3.28 10266 23321 0.53 3.55 2231 7311 0.17 4.57 1065 3648 0.08 4.72 1411 4154 0.09 Compound 13e λ: 254 nm, 8 nm 3.29 2036443 6802314 96.71 3.71 64999 158634 2.26 3.85 11659 43744 0.62 4.23 6974 18549 0.26 4.61 3917 10820 0.15 S24

Compound 13f λ: 254 nm, 1.2 nm 3.63 473290 12904.40 99.07 4.09 3657 79.34 0.61 5.40 1756 42.05 0.32 Compound 13g 2.77 1648 2996 0.17 3.29 720750 1737115 95.91 3.66 2482 8198 0.45 3.79 12091 33821 1.87 4.02 738 2295 0.13 4.19 2950 9408 0.52 4.30 1231 3206 0.18 4.46 1042 3276 0.18 4.59 822 4346 0.24 4.71 1837 6524 0.36 S25

Compound 21a 1.57 4222 11155 0.37 1.88 1140503 3013400 99.17 2.11 4097 14107 0.46 Compound 21b 1.94 27666 55311 2.10 2.03 1088159 2574064 97.90 S26

Compound 21c 2.15 1070208 2898094 97.39 2.22 40270 77709 2.61 Compound 21d 1.94 4940 11668 0.16 2.14 3149927 7216343 99.49 2.57 4939 13076 0.18 4.25 4180 12158 0.17 S27

Compound 21e 2.31 1175430 3049474 97.98 2.38 15495 49890 1.60 3.19 4909 13048 0.42 Compound 21f 2.13 1803563 4872410 95.93 2.34 6253 11785 0.23 2.56 74728 174389 3.43 2.79 8555 20560 0.40 S28

Compound 30a 2.01 1072796 2564552 99.32 2.21 7748 17641 0.68 Compound 30b 1.81 1417837 3745245 97.04 2.05 48354 114070 2.96 S29

Compound 30c λ: 254 nm, 1.2 nm 2.13 101569 2726.37 98.56 2.99 1765 39.74 1.44 Compound 30d 2.30 9562 25089 0.74 2.45 3712 7045 0.21 2.82 1170471 3282433 97.03 2.89 9633 20288 0.60 3.40 9159 28883 0.85 3.81 5912 19159 0.57 S30

Compound 30e λ: 215 nm, 8 nm 1.90 1072 2305 0.04 2.14 15216 42192 0.80 2.20 12281 40763 0.77 2.70 9931 33639 0.64 3.03 1794126 4988126 94.60 3.22 37589 144243 2.74 3.59 6361 18419 0.35 4.84 1059 2993 0.06 Compound 30f 2.66 1273417 3583133 100.00 S31

Compound 30g 2.95 1345667 3520429 100.00 Compound 37 λ: 254 nm, 1.2 nm 1.81 356 26.41 2.52 1.97 118 8.36 0.80 4.11 18319 1005.69 96.03 4.84 166 5.34 0.51 5.03 82 1.44 0.14 S32

Compound 42 λ: 254 nm, 1.2 nm 0.25 515 3.32 0.27 1.72 358 10.98 0.88 2.84 41583 1230.50 98.29 3.05 258 4.16 0.33 3.83 132 2.92 0.23 Compound 48a λ: 215 nm, 8 nm 2.32 2907907 7258972 95.49 2.38 192578 293999 3.87 2.82 15718 49082 0.65 S33

Compound 48b 2.44 1212137 2939952 96.83 2.97 11092 27594 0.91 3.95 7021 19516 0.64 4.07 8089 22979 0.76 4.27 7903 26205 0.86 Compound 48c 1.88 1168162 2849373 98.56 2.36 6438 23058 0.80 4.07 4860 18566 0.64 S34

Compound 48d λ: 254 nm, 1.2 nm 0.24 1563 13.94 0.26 4.54 171794 5264.72 99.19 5.31 648 17.40 0.33 5.42 245 11.72 0.22 Compound 48e 2.62 1359407 3813369 100.00 S35

Compound 48f 0.28 22745 50347 1.13 2.84 1359443 4422123 98.87 Compound 48g 2.35 986550 3190848 98.97 2.45 9236 33084 1.03 S36

Compound 48h 3.14 11474 35551 0.44 3.30 2019515 8030053 99.56 Compound 48i1 λ: 254 nm, 1.2 nm 2.36 492 17.45 0.24 2.42 524 10.19 0.14 4.87 278451 7245.08 99.40 5.97 517 16.19 0.22 S37

Compound 48i2 λ: 254 nm, 1.2 nm 0.23 343 3.93 0.30 4.16 1076 37.46 2.85 4.25 24016 1254.64 95.47 6.15 568 18.08 1.38 Compound 48j λ: 254 nm, 1.2 nm 1.67 222 12.51 0.19 1.72 1017 30.48 0.46 4.13 447 11.08 0.17 4.21 1044 20.09 0.30 4.29 129199 6617.49 98.89 S38

Compound 48k 2.51 11255 25546 0.24 2.71 3927654 10652574 99.60 3.16 5467 14685 0.14 4.12 1073 2824 0.03 Compound 48l 2.00 1667608 3879112 99.58 2.17 7704 16479 0.42 S39

Compound 48m λ: 215 nm, 8 nm 1.73 1234 2739 0.03 1.85 2475865 8491185 95.32 1.96 36142 374610 4.21 2.28 7631 21294 0.24 2.85 2764 18571 0.21 Compound 48n 2.64 938952 3499731 97.27 2.71 52627 94840 2.64 3.13 1200 3313 0.09 S40

Compound 48o 2.05 1963 4180 0.05 2.44 3617429 8449481 98.71 2.62 15080 61468 0.72 2.84 12130 40503 0.47 3.11 1864 4559 0.05 Compound 52a λ: 254 nm, 1.2 nm 0.23 1305 9.25 0.16 0.27 344 13.29 0.23 4.17 290 12.41 0.22 4.37 152945 5661.17 99.03 5.09 611 20.55 0.36 S41

Compound 52b λ: 254 nm, 1.2 nm 0.24 781 18.74 0.29 2.18 271 11.65 0.18 2.96 432 14.48 0.23 4.11 1090 25.15 0.39 4.34 198095 6313.32 98.90 Compound 52c λ: 220 nm, 1.2 nm 0.22 2483 18.84 0.18 0.34 1028 16.86 0.16 4.12 376105 10165.45 98.81 4.25 2036 37.23 0.36 4.94 4192 49.96 0.49 S42

Compound 52d λ: 254 nm, 1.2 nm 0.24 2617 17.22 0.27 3.59 247639 6052.09 95.00 3.67 3400 106.03 1.66 3.74 3487 73.75 1.16 5.27 4684 121.55 1.91 Compound 52e λ: 254 nm, 1.2 nm 2.15 826 21.74 0.16 3.66 1235 71.23 0.42 3.84 545756 13938.79 94.62 4.17 11928 285.50 1.84 5.27 17856 462.46 2.96 S43

Compound 52f λ: 254 nm, 1.2 nm 0.24 3052 22.40 0.29 2.30 2390 45.38 0.58 2.36 486 16.76 0.21 3.44 909 23.28 0.30 3.67 756 20.19 0.26 3.82 320067 7605.60 97.43 4.03 1251 35.52 0.46 4.38 543 19.60 0.25 5.32 702 17.47 0.22 S44

S6. Figure S1. Superposition of sildenafil and vardenafil in the different crystal complexes with PDE5: 1TBF (sildenafil, orange) 1, 2H42 (sildenafil, pink) 2, 1UDT (sildenafil, blue) 3 and 3B2R (vardenafil, green) 4. S45

S7. Table S1. PDE5A and HDACs biochemical activities as pic 50. Cpd PDE5A HDAC1 HDAC2 HDAC3- HDAC6 pic 50 pic 50 pic 50 NCOR2 pic 50 pic 50 13a 8.52 4.98 <4.70 5.63 13b 8.70 5.96 5.33 6.44 13c 8.52 8.10 6.93 7.44 * 6.57 13d 8.70 5.87 5.16 7.05 13e 9.30 6.39 5.71 7.06 13f 9.22 7.24 6.47 7.27 * 7.23 13g 9.00 6.45 5.88 7.08 21a 7.36 5.15 <4.70 4.98 21b 7.66 5.22 <4.70 4.90 21c 7.70 5.11 4.76 5.80 21d 8.30 7.17 6.31 7.51 6.36 21e 7.77 6.93 6.15 6.15 21f 8.00 7.60 6.78 7.37 6.23 30a 7.19 5.23 <4.70 5.96 30b 7.62 6.22 5.69 5.97 30c 7.92 4.89 <4.70 5.36 30d 7.40 5.03 <4.70 5.00 30e 7.13 6.35 5.73 5.72 S46

30f 7.96 5.61 5.01 5.63 30g 7.64 6.25 5.65 6.71 37 8.00 7.18 6.36 6.43 42 8.30 5.19 <4.70 <4.70 48a 7.72 <4.70 <4.70 <4.70 48b 7.34 5.32 <4.70 5.67 52a 7.89 5.62 <4.70 5.63 52b 6.91 5.81 <4.70 5.75 48c 8.15 7.85 7.05 6.42 48d 8.15 7.20 6.47 7.29 5.90 48e 7.24 5.73 5.19 6.40 48f 6.92 <4.70 <4.70 <4.70 48g 7.24 <4.70 <4.70 <4.70 48h 7.42 5.81 <4.70 6.46 48i1 7.89 6.17 <4.70 6.29 48i2 7.66 6.46 <4.70 7.24 48j 6.78 5.45 4.86 6.38 7 7.22 6.51 6.31 6.49 7.04 7a 7.47 6.65 6.14 6.55 6.84 7b 7.35 6.49 5.91 6.62 6.90 48k 7.41 5.58 4.85 5.72 48l 6.68 5.12 <4.70 5.02 48m 6.52 <4.70 <4.70 <5.00 S47

48n 7.77 6.26 5.73 6.89 48o 7.70 5.06 <4.70 5.27 52c 7.15 5.16 <4.70 5.29 52d 8.40 6.45 5.73 7.10 52e 8.40 5.63 <4.70 6.06 52f 8.30 <4.70 <4.70 5.25 * HDAC3_NCOR2 values obtained at BPS (https://bpsbioscience.com/) BIBLIOGRAPHY (1) Bischoff, E. Int. J. Impot. Res. 2004, 16 Suppl 1, S11 S14. (2) Wang, H.; Liu, Y.; Huai, Q.; Cai, J.; Zoraghi, R.; Francis, S.H.; Corbin, J.D.; Robinson, H.; Xin, Z.; Lin, G.; Ke, H. J. Biol. Chem. 2006, 281, 21469-21479. (3) Sung, B.-J.; Hwang, K. Y.; Jeon, Y. H.; Lee, J. I.; Heo, Y.-S.; Kim, J. H.; Moon, J.; Yoon, J. M.; Hyun, Y.-L.; Kim, E.; Eum, S. J.; Park, S.-Y.; Lee, J.-O.; Lee, T. G.; Ro, S.; Cho, J. M. Nature 2003, 425 (6953), 98 102. (4) Wang, H.; Ye, M.; Robinson, H.; Francis, S. H.; Ke, H. Mol. Pharmacol. 2008, 73 (1), 104 110. S48