Total Synthesis and Biological Evaluation of. Cortistatins A and J and Analogues Thereof

Transcription

1 Total Synthesis and Biological Evaluation of Cortistatins A and J and Analogues Thereof K. C. Nicolaou,* Xiao-Shui Peng, Ya-Ping Sun, Damien Polet, Bin Zou, Chek Shik Lim, and David Y.-K. Chen* Contribution from Chemical Synthesis Laboratory@Biopolis, Institute of Chemical and Engineering Sciences (ICES), Agency for Science, Technology and Research (A*STAR), 11 Biopolis Way, The elios Block, #03 08, Singapore kcn@scripps.edu; david_chen@ices.a-star.edu.sg I) Experimental Section Supporting Information Available II) III) IV) V) Abbreviations Materials and thods for Cytotoxocity Assay References 1 and 13 C NMR Spectra of Compounds I) Experimental Section Experimental Data for Compounds General Procedures. All reactions were carried out under a nitrogen or argon atmosphere with dry solvents under anhydrous conditions, unless otherwise noted. Dry tetrahydrofuran (TF), methylene chloride (C 2 Cl 2 ), and toluene were obtained by passing commercially available pre-dried, oxygen-free

2 formulations through activated alumina columns. thanol (), N,N -dimethylformamide (DMF), dimethylsulfoxide (DMS), dioxane and acetonitrile (CN) were purchased in anhydrous form and used without further purification. Acetone, water, ethyl acetate (EtAc) and hexanes were purchased at the highest commercial quality and used without further purification, unless otherwise stated. Reagents were purchased at the highest commercial quality and used without further purification, unless otherwise stated. Yields refer to chromatographically and spectroscopically ( 1 NMR) homogeneous materials, unless otherwise stated. Reactions were monitored by thin-layers chromatography (TLC) carried out on 0.25 mm E. rck silica gel plates (60F-254) using UV light as visualizing agent and an ethanolic solution of ammonium molybdate and anisaldehyde and heat as developing agents. E. rck silica gel (60, particle size mm) was used for flash column chromatography. NMR spectra were recorded on Bruker AV-600 instrument and calibrated using residual undeuterated solvent as an internal reference. The following abbreviations were used to explain the multiplicities: s = singlet, d = doublet, t = triplet, q = quartet, quint = quintet, m = multiplet, pent = pentet, hex = hexet, br = broad. IR spectra were recorded on a Perkin-Elmer Spectrum ne FTIR spectrometer with diamond ATR accessory. ptical rotation ([ α] D ) were recorded on a Perkin-Elmer Model 341 polarimeter at 25 C using thermostable optical glass cell (100mm path length). lting points (m.p.) are uncorrected and were recorded on a Buchi B-540 melting point apparatus. ighresolution mass spectra (RMS) were recorded on an Agilent ESI TF (time of flight) mass spectrometer at 3500 V emitter voltage. TES ether 18: To a stirred solution of alcohol 17 1 (10 g, 79 mmol) in DMF (30 ml) was added sequentially imidazole (10.7 g, 158 mmol) and TESCl (14.3 g, 95 mmol) at room temperature. The TES reaction mixture was stirred for 6 h before it was quenched with NaC 3 (30 ml, sat. aq.) and extracted with EtAc (4 30 ml). The combined organic layers were dried 18 (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 10:1) afforded TES ether 18 (14.3 g, 81%) as a colorless oil. 18: R f = 0.30 (silica gel, hexanes:etac 30:1); IR (film) ν max 2978, 2746, 1730, 1260, 1245 cm 1 ; 1 NMR S2

3 (600 Mz, CDCl 3 ): δ = 7.04 (m, 1 ), 4.36 (t, J = 2.4 z, 2 ), 2.41 (m, 4 ), 2.00 (t, J = 6.6 z, 2 ), 0.97 (t, J = 8.4 z, 9 ), 0.64 ppm (q, J = 15.6 z, 6 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 199.1, 144.1, 138.3, 59.6, 38.4, 25.6, 22.9, 6.7, 4.4 ppm; RMS (ESI): calcd for C SiNa + [M + Na + ] , found Acetylene 20: To a stirred solution of enone 18 (5 g, 22.3 mmol) in TF (50 ml) at 0 C was added a solution 20 of lithium trimethylsilylacetylide [prepared by treatment of a solution of trimethylsilylacetylene (3.3 g, 33.5 mmol) in TF (30 ml) with of n-buli (1.6 M in hexanes, 21 ml, 33.5 mol) at 0 C]. The reaction mixture was then allowed to warm up to room temperature and stirred for 2 h before it was quenched with N 4 Cl (20 ml, sat. aq.) and extracted with EtAc (4 50 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. The crude residue was dissolved in TF (20 ml) and stirred with TBAF (1 M in TF, 30 ml, 30 mmol) at room temperature for 30 min. before it was concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) afforded acetylene 20 (2.4 g, 72%) as a colorless oil. 20: R f = 0.25 (silica gel, hexanes:etac 2:1); IR (film) ν max 3406, 2978, 2647, 1675, 1639, 1260, 1247 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 5.84 (t, J = 4.2 z, 1 ), 4.63 (d, J = 12.0 z, 1 ), 4.11 (d, J = 12.0 z, 1 ), 4.10 (brs, 1 ), 3.05 (brs, 1 ), 2.53 (s, 1 ), 2.11 (m, 1 ), 2.04 (m, 3 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 137.0, 129.2, 87.2, 72.1, 67.9, 65.7, 38.3, 25.1, 18.7 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Enynone 21: To a mixture of acetylene 20 (2.0 g, 13 mmol) were added enol triflate 12 [freshly prepared from 1,4-cyclohexadione (6.7 g, 60 mmol), Tf 2 (2.1 g, 72 mmol) and Et 3 N (17.3 ml, 120 mmol) in C 2 Cl 2 (150 ml)], Pd(PPh 3 ) 4 (750 mg, 0.65 mmol), CuI (247 mg, 1.3 mmol) in DMF (88 ml) and Et 3 N (5.5 ml, 40 mmol) at room temperature. The reaction mixture was stirred for 1 h before it was quenched with 2 (50 ml) and 21 extracted with EtAc (3 60 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) S3

4 afforded enynone 21 (2.6 g, 82%) as a yellow oil. 21: R f = 0.24 (silica gel, hexanes:etac 1:1); IR (film) ν max 3006, 2994, 1732, 1655, 1275 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.18 (s, 1 ), 5.87 (t, J = 3.6 z, 1 ), 4.61 (d, J = 12.0 z, 1 ), 4.30 (s, 1 ), 4.13 (d, J = 4.2 z, 1 ), 3.11 (s, 1 ), 2.44 (t, J = 6.0 z, 2 ), 2.39 (t, J = 6.6 z, 1 ), 2.13 (m, 1 ), 2.06 (m, 5 ), 1.81 (m, 1 ), 1.69 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 199.1, 143.5, 136.6, 132.6, 129.7, 103.4, 82.8, 68.3, 65.9, 38.0, 37.2, 30.4, 25.1, 22.5, 18.7 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Enone enal 22: To a stirred solution of alcohol 21 (2.5 g, 10 mmol) in C 2 Cl 2 (50 ml) were added NaC 3 (5 g, 60 mmol) and DMP (6.4 g, 15 mmol) at room temperature. The reaction mixture was stirred for 30 min before it was quenched with NaC 3 (50 ml, sat. aq.), Na 2 S 2 3 (30 ml, sat. aq.), and extracted with C 2 Cl 2 (4 100 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 4:1) afforded enone enal (1.76 g, 71%) as a colorless oil. 22: R f = 0.30 (silica gel, hexanes:etac 6:1); IR (film) ν max 3209, 1635, 1666, 1260, 1236 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 9.36 (s, 1 ), 6.86 (t, J = 3.6 z, 1 ), 6.08 (s, 1 ), 4.54 (s, 1 ), 2.35 (m, 6 ), 2.16 (dd, J = 10.2, 2.4 z, 1 ), 1.95 (t, J = 5.4 z, 2 ), 1.83 (m, 2 ), 1.76 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 198.5, 194.1, 153.2, 142.8, 141.8, 132.7, 101.5, 83.3, 65.8, 37.2, 36.4, 30.3, 26.6, 22.4, 18.6 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found ydroxy enone enal 23: To a stirred solution of alkyne 22 (1.6 g, 0.65 mmol) in /EtAc (2:3, 350 ml) was added Pd/C (10% wt/wt, 266 mg, 0.25 mmol) at room temperature. The resulting mixture 23 was stirred under 2 (1 atm) for 25 min before it was filtered and washed through with EtAc (3 80 ml). The combined organic fractions were concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 3:1) afforded hydroxy enone enal 23 (0.99 g, 61%) as a colorless oil. 23: R f = 0.30 (silica gel, hexanes:etac 4:1); IR (film) ν max 3006, 1733, 1462, 1275, 1260 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): S4

5 δ = 9.37 (s, 1 ), 6.84 (t, J = 4.2 z, 1 ), 5.86 (t, J = 1.2 z, 1 ), 3.81 (brs, 1 ), 2.38 (m, 3 ), 2.33 (t, J = 4.2 z, 2 ), 2.29 (t, J = 7.2 z, 2 ), 2.21 (m, 1 ), 2.02 (m, 1 ), 1.99 (m, 2 ), 1.92 (m, 1 ), 1.79 (m, 2 ), 1.64 ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 199.7, 195.7, 166.2, 154.4, 144.7, 125.6, 71.2, 37.3, 36.3, 32.7, 31.7, 29.9, 26.8, 22.7, 18.3 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Dienone 26: thod A: To a stirred solution of hydroxy enone enal 23 (51 mg, 0.21 mmol) in (5.0 ml) was added a solution of K (10% in, 0.5 ml, 0.9 mmol) at room temperature. The resulting 26 mixture was stirred at 101 o C for 12 h before it was concentrated in vacuo to remove most of the. The residue was taken up with water (10 ml) and extracted with EtAc (3 30 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded dienone 26 (14.7 mg, 31%) as a colorless oil. 26: R f = 0.25 (silica gel, hexanes:etac 8:1); IR (film) ν max 2707, 2318, 1734, 1426, 1260, 1255 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.90 (s, 1 ), 5.92 (t, J = 3.6 z, 1 ), 2.55 (dd, J = 15.6, 4.6 z, 1 ), 2.36 (m, 1 ), 2.24 (m, 3 ), 2.01 (m, 4 ), 1.91 (m, 5 ), 1.80 (m, 1 ), 1.62 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 198.3, (2), 132.7, 132.0, 82.3, 80.5, 41.4, 39.4, 38.1, 33.6, 26.3 (2), 20.3, 19.0 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found thod B: To a stirred solution of hydroxy enone enal 23 (43 mg, 0.17 mmol) in toluene (6.0 ml) was added piperidine (1.68 µl, mmol) at room temperature. The resulting mixture was refluxed for 22 h before it was concentrated in vacuo to remove most of the toluene. The residue was taken up with water (8 ml) and extracted with EtAc (3 25 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded dienone 26 (8.4 mg, 21%) as a colorless oil. thod C: To a stirred solution of hydroxy enone enal 23 (400 mg, 1.63 mmol) in dioxane (16.0 ml) was added K 2 C 3 (pre-dried, 270 mg, 1.96 mmol) at room temperature. The resulting mixture was S5

6 heated at 125 C for 16 h before it was concentrated in vacuo to remove most of the dioxane. The residue was taken up with water (15 ml) and extracted with EtAc (3 50 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded dienone 26 (185 mg, 50%) as a colorless oil. thod D: To a stirred solution of hydroxy enone enal 23 (65 mg, 0.26 mmol) in toluene (8.0 ml) was added PPTS (6.5 mg, mmol) at room temperature. The resulting mixture was refluxed for 14 h before it was concentrated in vacuo to remove most of the toluene. The residue was taken up with water (10 ml) and extracted with EtAc (3 40 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded dienone 26 (13 mg, 22%) as a colorless oil. thod E: To a stirred solution of hydroxy enone enal 23 (62 mg, 0.25 mmol) in toluene (8.0 ml) was added PTSA (4.7 mg, mmol) at room temperature. The resulting mixture was refluxed for 14 h before it was concentrated in vacuo to remove most of the toluene. The residue was taken up with water (10 ml) and extracted with EtAc (3 40 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded dienone 26 (9.7 mg, 17%) as a colorless oil. Trienone 27: thod A (Saegusa oxidation): To a stirred solution of dienone 26 (90 mg, 0.39 mmol) in TF (4.0 ml) was added Et3N (0.11 ml, 0.8 mmol) at room temperature. The resulting solution was cooled to 78 C and TMSTf (0.087 ml, 0.48 mmol) was added dropwise before the reaction mixture was warmed to 0 C and stirred for 1.5 h. The reaction mixture was quenched with NaC 3 (10 ml, sat. aq.), stirred at room temperature for 15 min, 27 and extracted with Et 2 (3 25 ml). The combined organic layers were washed with brine (20 ml), dried (Na 2 S 4 ) and concentrated in vacuo to afford the crude TMS enol ether. To a stirred solution of crude TMS enol ether (obtained above) in C 3 CN (10 ml) was added Pd(Ac) 2 (135 mg, 0.6 mmol) at room temperature. The reaction mixture was stirred for 15 h before it S6

7 was concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded trienone 27 (38 mg, 43%) as a colorless oil. 27: R f = 0.25 (silica gel, hexanes:etac 8:1); IR (film) ν max 2930, 2863, 1657, 1630, 1457, 1260, 1245 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 7.05 (s, 1), 6.94 (ddd, J = 8.4, 6.6, 2.4 z, 1 ), 6.21 (ddd, J = 10.2, 3.6, 0.6 z, 1 ), 5.94 (t, J = 3.6 z, 1 ), 2.98 (dt, J = 18.0, 1.8 z, 1 ), 2.68 (dd, J = 18.0, 6.6 z, 1 ), 2.40 (dd, J = 10.8, 2.4 z, 1 ), 2.26 (m, 2 ), 2.05 (m, 1 ), 1.94 (m, 4 ), 1.88 (m, 1 ), 1.64 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 185.7, 145.5, 140.0, 138.1, 132.0, 131.6, 130.3, 81.5, 80.2, 42.0, 38.3, 34.8, 33.6, 26.2, 20.4 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found thod B (IBX MP): To a stirred solution of dienone 26 (20 mg, mmol) in TF (1.0 ml) was added Et 3 N (0.024 ml, mmol) at room temperature. The resulting solution was cooled to 78 C and TMSTf (0.019 ml, mmol) was added dropwise before the mixture was warmed to 0 C and stirred for 1.5 h. The reaction mixture was quenched with NaC 3 (3 ml, sat. aq.), stirred at room temperature for 15 min, and extracted with Et 2 (3 10 ml). The combined organic layers were washed with brine (5 ml), dried (Na 2 S 4 ) and concentrated in vacuo to afford the crude TMS enol ether. To a stirred solution of the crude TMS enol ether (obtained above) in DMS (1 ml) was added a solution of IBX MP (0.4 M in DMS, 1.3 ml, 0.52 mmol) at room temperature. The reaction mixture was stirred for 6 h before it was quenched with NaC 3 (5 ml, sat. aq.) and stirred for 15 min. The resulting mixture was extracted with EtAc (4 10 ml), and the combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 6:1) afforded trienone 27 (9.0 mg, 45% over the two steps) as a colorless oil. Keto carbamate 28: To a stirred solution of trienone 27 (20 mg, 0.09 mmol) in TF (1.0 ml) at 78 C was added a solution of AllocN()Li [prepared by adding a solution of n-buli (2.5 M in hexane, 0.18 ml, 0.45 mmol) to a stirred solution of AllocNC 3 (61 mg, 0.53 mmol) in TF (3.0 ml) at 78 C and stirred for 1 h]. The resulting mixture was stirred for 15 min before it was quenched with N 4 Cl (1.5 ml, sat. aq.) at 78 C. The resulting biphasic mixture was warmed to room temperature and S7

8 extracted with EtAc (5 5 ml). The combined organic layers were AllocN 28 washed with brine (10 ml), dried (Na 2 S 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) afforded keto carbamate 28 (21 mg, 70%) as a yellow oil. 28: R f = 0.25 (silica gel, hexanes:etac 2:1); IR (film) ν max 2925, 1698, 1577, 1456, 1260, 1170 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.81 (s, 1 ), 5.94 (m, 2 ), 5.22 (dd, J = 18.0, 10.2 z, 2 ), 4.60 (t, J = 5.4 z, 2 ), 2.88 (s, 3 ), 2.58 (d, J = 4.4 z, 1 ), (m, 2 ), (m, 2 ), (m, 7 ), ppm (m, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 197.7, 155.6, 140.2, 133.6, 133.2, (2), 117.5, 81.5, 79.6 (2), 66.1, 42.9, 38.2 (2), 33.4 (2), 29.7, 26.4, 20.1 ppm; RMS (ESI): calcd for C20 25 N 4 Na + [M + Na + ] , found Triene 32: To a stirred solution of keto carbamate 28 (20 mg, 0.06 mmol) and PhNTf 2 (42 mg, 0.12 mmol) in TF (2.0 ml) was added KMDS (0.5 M in toluene, 0.36 ml, 0.18 mmol) at 78 C. The resulting mixture was warmed to 0 C and stirred for 0.5 h before it was N 32 quenched with N 4 Cl (2.0 ml, sat. aq.) and extracted with EtAc (5 10 ml). The combined organic layers were washed with brine (5 ml), dried (Na 2 S 4 ), and concentrated in vacuo. Filtration through a plug of silica and elution with (hexane:etac 3:1, 30 ml) afforded the semi-purified enol triflate of 31, which was used directly without further purification. To a stirred solution of semi-purified enol triflate (obtained above) in C 2 Cl 2 (10.0 ml) were added acetic acid (82 μl, 1.4 mmol) and n-bu 3 Sn (32 μl, 0.12 mmol) at room temperature. The reaction mixture was degassed three times (freeze-pump-thaw cycle) before Pd(PPh 3 ) 2 Cl 2 (2.1 mg, mmol) was added at room temperature. The resulting mixture was stirred for 1 h before it was concentrated in vacuo. Flash column chromatography (silica gel, EtAc: 3:1) afforded triene 32 (3.8 mg, 27% over two steps from 28) as a colorless oil. 32: R f = 0.28 (silica gel, EtAc: 3:1); IR (film) ν max 3327, 2925, 2862, 1563, 1442, 1402, 1335, 1275, 1133, 1044 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.11 (d, J = 9.6 z, 1 ), 5.95 (dd, J = 9.6, 5.4 z, 1 ), 5.81 (s, 1 ), 5.51 (t, J = 4.8 z, 1 ), 3.42 (t, S8

9 J = 4.8 z, 1 ), 2.95 (bs, 1 ), 2.54 (s, 3 ), (m, 5 ), (m, 1 ), (m, 5 ), ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 140.9, 139.8, 128.2, 127.2, 123.9, 123.0, 80.0, 78.4, 53.9, 40.6, 39.2, 34.2, 34.0, 33.8, 25.7, 20.7 ppm; RMS (ESI): calcd for C16 21 NNa + [M + Na + ] , found Dimethyl amine 33: To a stirred solution of methyl amine 32 (3.0 mg, mmol) in (0.9 ml) at room temperature were added formaldehyde (37% aq., 2.0 µl, mmol) and NaCNB 3 (4.4 mg, mmol). The resulting mixture was stirred for 1 h before it was 2 N 33 quenched with Na (1.0 M aq., 1.0 ml). The resulting mixture was stirred for 15 min before it was extracted with EtAc (3 4 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Preparative TLC purification (silica gel, EtAc: 3:1) afforded dimethyl amine 33 (1.8 mg, 56%) as a white amorphous solid. 33: R f = 0.32 (silica gel, EtAc: 3:1); IR (film) ν max 2989, 1736, 1463, 1372, 1275, 1265, 1044 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.12 (d, J = 9.6 z, 1 ), 5.83 (dd, J = 9.6, 5.4 z, 1 ), 5.79 (s, 1 ), 5.48 (t, J = 4.2 z, 1 ), 2.88 (bs, 1 ), 2.33 (bs, 6 ), (m, 2 ), (m, 2 ), (m, 2 ), ppm (m, 6 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 141.3, 140.6, 128.6, 128.1, 123.0, 122.2, 80.2, 79.2, 60.5, 44.3, 41.0, 39.1, 34.2, 33.7, 25.7, 20.7 ppm; RMS (ESI): calcd for C N + [M + + ] , found ydroxy ketone 29: To a stirred solution of ketone 28 (40 mg, 0.12 mmol) in TF (2.0 ml) was added KMDS (0.5 M in toluene, 0.72 ml, 0.36 mmol) at 78 C. The resulting mixture was stirred for 0.5 h before a solution of Davis oxaziridine (60 mg, 0.24 mmol) in TF (1.0 ml) was added dropwise. The resulting mixture was stirred for 1 h AllocN before it was quenched with N 4 Cl (2.0 ml, sat. aq.) and extracted with 29 EtAc (3 15 ml). The combined organic layers were washed with brine (5 ml), dried (Na 2 S 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 3:2) afforded hydroxy ketone 29 (25 mg, 60%) as a yellow amorphous solid. 29: R f = 0.40 (silica gel, hexanes:etac 1:1); IR (film) ν max 3453, 2940, 1686, 1572, 1449, 1275, 1159 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.01 (d, S9

10 J = 9.6 z, 1 ), 5.95 (ddd, J = 16.2, 10.8, 5.4 z, 1 ), 5.89 (s, 1 ), 5.33 (dd, J = 16.8, 6.0 z, 1 ), 5.22 (d, J = 10.8 z, 1 ), 5.27 (s, 1 ), 4.92 (s, 1 ), 4.60 (s, 2 ), 2.98 (s, 3 ), 2.48 (td, J = 12.6, 8.4 z, 2 ), 2.31 (m, 1 ), 2.21 (s, 1 ), (m, 5 ), 1.76 (dd, J = 11.4, 4.2 z, 1 ), 1.69 (m, 1 ), 1.61 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 208.2, 156.4, 143.3, 133.0, 130.9, 125.2, 118.0, 117.3, 83.3, 82.5, 72.2, 66.1, 35.0, 33.9, 30.8, 30.6, 24.6 ppm; RMS (EI): calcd for C N , found Nitrobenzoate 29a: To a stirred solution of DCC (7.7 mg, mmol), 4-nitrobenzoic acid (6.2 mg, mmol) and 4-DMAP (0.61 mg, mmol) in C 2 Cl 2 (0.5 ml) was added a solution of alcohol R AllocN 29a R = 4-Nitrobenzoate 29 (9.0 mg, mmol) in C 2 Cl 2 (0.5 ml) at room temperature. The resulting mixture was stirred for 2 h before it was quenched with NaC 3 (5 ml, sat. aq.) and extracted with Et 2 (3 10 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) afforded nitrobenzoate 29a (9.0 mg, 71% yield) as a white solid. 29a: R f = 0.60 (silica gel, hexanes:etac 1:1); m.p. = 145 C (decomp. C 2 Cl 2 /hexanes); IR (film) ν max 2941, 1735, 1697, 1628, 1573, 1528, 1347, 1268 cm 1 ; 1 NMR (600 Mz, CD 3 CN, 65 C): δ = 8.32 (d, J = 9.0 z, 2 ), 8.19 (d, J = 9.0 z, 2 ), 6.98 (s, 1 ), 6.12 (t, J = 3.9 z, 1 ), 5.88 (brs, 1 ), 5.74 (brs, 1 ), (m, 2 ), 4.70 (brs, 1 ), 4.52 (d, J = 4.8 z, 2 ), 2.88 (s, 3 ), 2.58 (ddd, J = 15.0, 11.4, 3.6 z, 2 ), (m, 3 ), 2.30 (quint, J = 3.6 z, 2 ), (m, 3 ), ppm (m, 2 ); 13 C NMR (150 Mz, CD 3 CN, 65 C): δ = 192.7, 165.5, 157.5, 152.8, 142.1, 140.2, 136.8, 136.7, 135.1, 133.0, 132.5, 125.6, 125.5, 83.3, 80.4, 77.4, 71.2, 67.6, 67.5, 45.1, 39.9, 35.1, 27.9, 21.5 ppm; RMS (EI): calcd for C N 2 8 Na + [M + Na + ] , found Diol 30: To a stirred solution of 4 NB(Ac) 3 (70 mg, mmol) in C 3 CN (0.7 ml) was added Ac (0.7 ml) at room temperature. The resulting mixture was stirred for 15 min before a solution of the ketone 29 (12 mg, mmol) in C 3 CN (1.0 ml) was added at room temperature and the resulting mixture was stirred for 1 h before it was quenched with potassium sodium tartrate (5 ml, S10

11 sat. aq.), neutralized with NaC 3 (10 ml, sat. aq.), and extracted with EtAc (3 15 ml). The combined organic layers were dried (Na 2 S 4 ) and AllocN 30 concentrated in vacuo. Preparative TLC purification (silica gel, hexanes:etac 2:1) afforded diol 30 (3.0 mg, 25%) as a colorless oil. 30: R f = 0.25 (silica gel, hexanes:etac 1:1); IR (film) ν max 3404, 2936, 1685, 1450, 1405, 1159 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.04 (s, 1 ), 5.95 (dddd, J = 15.6, 10.2, 5.4, 3.0 z, 1 ), 5.46 (brs, 1 ), 5.32 (d, J = 15.0 z, 1 ), 5.24 (d, J = 10.8 z, 1 ), 4.60 (s, 2 ), 4.35 (d, J = 5.4 z, 1 ), 4.19 (td, J = 6.6, 1.8 z, 1 ), 3.90 (td, J = 6.6, 3.0 z, 1 ), 2.94 (s, 3 ), (m, 2 ), (m, 2 ), (m, 5 ), ppm (m, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 142.0, 139.6, 132.8, 121.9, 119.6, 117.7, 81.9, 79.7, 72.8, 67.6, 66.4, 60.3, 39.7, 37.8, 34.2, 32.3, 31.8, 25.4, 20.5 ppm; RMS (EI): calcd for C N , found Epoxy ketone 34: To a stirred solution of trienone 27 (10 mg, mmol) in C 2 Cl 2 (2.5 ml) were added t-bu (5.5 M solution in decane, 32 μl, 0.18 mmol) and DBU (20 μl, 0.13 mmol) at 0 C. The resulting mixture was warmed to room temperature and stirred for 5 h before it was quenched with Na 2 S 2 3 (2 ml, sat. aq.) and NaC 3 (5 ml, sat. 34 aq.). The resulting mixture was stirred for 15 min and extracted with Et 2 (3 20 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 3:1) afforded epoxy ketone 34 (7.7 mg, 72% yield) as a white solid. 34: R f = 0.30 (silica gel, hexanes:etac 6:1); m.p. = C (C 2 Cl 2 /hexanes); IR (film) ν max 2937, 1677, 1627, 1577, 1450, 1289, 1249 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 7.08 (s, 1 ), 5.98 (t, J = 3.6 z, 1 ), 3.69 (t, J = 3.6 z, 1 ), 3.46 (d, J = 4.2 z, 1 ), 2.56 (dd, J = 9.0, 3.6 z, 1 ), 2.51 (m, 2 ), 2.26 (m, 2 ), 2.00 (m, 2 ), 1.94 (m, 2 ), 1.89 ppm (m, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 192.1, 140.2, 137.0, 134.0, 133.9, 79.8, 78.9, 54.6, 53.6, 44.3, 38.8, 33.5, 32.6, 26.4, 20.2 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Epoxy alcohols 35 and 36: To a stirred solution of epoxy ketone 34 (10 mg, mmol) in (3.0 ml) was added CeCl (46 mg, 0.12 mmol) at 0 C. The resulting mixture was stirred for 10 S11

12 min before a solution of NaB 4 (1.6 mg, mmol) in (0.3 ml) was added dropwise. The resulting mixture was stirred for 10 min before it was quenched with NaC 3 (5 ml, sat. aq.) and stirred for 15 min. The mixture was extracted with EtAc (3 15 ml), and the combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Preparative TLC purification (silica gel, hexanes:etac 2:1) afforded epoxy alcohols 35 (4.0 mg, 40%) and 36 (4.0 mg, 40%) as white solids. 35: R f = 0.22 (silica gel, hexanes:etac 4:1); IR (film) ν max 3428, 2936, 2167, 2042, 1942, 1275, 1260 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.16 (s, 1 ), 5.50 (t, J = 4.2 z, 1 ), 4.70 (s, 1 ), 3.50 (s, 1 ), 3.41 (t, J = 4.2 z, 1 ), 3.28 (dd, J = 4.2, 2.4 z, 1 ), 2.50 (d, J = 14.4 z, 1 ), 2.39 (dd, J = 14.4, 2.4 z, 1 ), 2.33 (m, 1 ), 2.17 (m, 2 ), 2.01 (m, 1 ), 1.93 (m, 2 ), 1.91 (d, J = 3.0 z, 2 ), 1.84 (m, 1 ), 1.58 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 141.8, 139.8, 126.2, 123.8, 80.4, 78.7, 66.9, 53.3, 52.6, 44.3, 38.6, 33.9, 32.6, 25.5, 20.5 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found : R f = 0.25 (silica gel, hexanes:etac 4:1); m.p. = C (C 2 Cl 2 /hexanes); IR (film) ν max 3428, 2935, 2164, 2042, 1942, 1275, 1260 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.20 (s, 1 ), 5.50 (t, J = 3.6 z, 1 ), 4.72 (s, 1 ), 3.49 (dd, J = 6.6, 3.0 z, 1 ), 3.44 (t, J = 3.0 z, 1 ), 2.41 (m, 3 ), 2.17 (m, 2 ), 2.02 (m, 1 ), 1.89 (m, 6 ), 1.59 ppm (m, 1 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 142.3, 139.9, 126.5, 123.7, 80.1, 79.7, 68.3, 54.2, 52.9, 44.8, 38.4, 33.9, 32.5, 25.4, 20.5 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Dimethyl amines 38 and 41: To a stirred solution of epoxy alcohol 36 (8.0 mg, mmol) in 2 N (2.0 M solution in TF, 2.0 ml) was added Ti(i-Pr) 4 (48.0 μl, mmol) at room temperature. The resulting mixture was heated 2 N to 80 C in a sealed tube for 5 h and then 2 N quenched with NaC 3 (5 ml, sat. aq.) and stirred for 15 min before it was extracted with EtAc (3 15 ml). The combined organic layers were S12

13 dried (Na 2 S 4 ) and concentrated in vacuo. Preparative TLC purification (silica gel, ) afforded dimethyl amines 38 (4.2 mg, 44%) and 41 (3.3 mg, 35%) as white amorphous solids. 38: R f = 0.20 (silica gel, ); IR (film) ν max 3367, 2934, 2864, 1457, 1275, 1261, 1049 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.20 (d, J = 2.4 z, 1 ), 5.49 (t, J = 4.2 z, 1 ), 4.11 (d, J = 9.0 z, 1 ), 3.35 (t, J = 9.6 z, 1 ), 2.41 (ddd, J = 13.8, 10.2, 3.6 z, 1 ), 2.29 (s, 6 ), (m, 3 ), 2.07 (ddd, J = 12.6, 9.6, 7.2 z, 1 ), 1.97 (dd, J = 12.6, 3.6 z, 1 ), (m, 4 ),1.80 (td, J = 10.8, 6.6 z, 1 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 140.5, 139.2, 122.6, 119.8, 80.7, 80.0, 74.1, 73.8, 62.1, 40.9, 40.1, 38.3, 34.1, 29.3, 25.4, 20.7 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found : R 367, 2934, 2864, 1457, 1275, 1261, 1049 cm 1 ; 1 f = 0.23 (silica gel, ); IR (film) ν max 3 NMR (600 Mz, CDCl 3 ): δ = 6.02 (d, J = 1.8 z, 1 ), 5.37 (t, J = 3.6 z, 1 ), 4.30 (m, 1 ), 4.29 (d, J = 7.8 z, 1 ), 2.99 (t, J = 8.4 z, 1 ), 2.46 (s, 6 ), (m, 2 ), 2.08 (t, J = 9.0 z, 1 ), (m, 3 ), (m, 4 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 145.3, 140.0, 120.7, 117.5, 81.8, 79.6, 69.5, 66.1, 63.7, 43.5, 39.9, 39.2, 37.9, 34.2, 25.4, 20.6 ppm; RMS (ESI): calcd for C [M + + ] , found TBS ether 37: To a stirred solution of epoxy alcohol 36 (7 mg, mmol) in C 2 Cl 2 (1.0 ml) were added 2,6-lutidine (10 μl, mmol) and TBSTf (13 μl, mmol) at 0 C. The resulting TBS 37 mixture was stirred for 1 h before it was quenched with NaC 3 (5 ml, sat. aq.). The resulting mixture was stirred for 15 min and extracted with Et 2 (3 10 ml), and the combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 9:1) afforded TBS ether 37 (8.2 mg, 80% yield) as a colorless oil. 37: R f = 0.30 (silica gel, hexanes:etac 10:1); IR (film) ν max 2930, 1472, 1463, 1361, 1258, 1061, 836 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 5.98 (s, 1 ), 5.43 (t, J = 3.6 z, 1 ), 4.53 (s, 1 ), 3.38 (dd, J = 6.0, 3.6 z, 1 ), 3.15 (dd, J = 4.2, 2.4 z, 1 ), 2.49 (dd, J = 15.0, 2.4 z, 1 ), 2.22 (dd, J = 15.0, 3.6 z, 1 ), (m, 3 ), 1.99 (m, 1 ), S13

14 (m, 5 ), 1.79 (dd, J = 11.4, 5.4 z, 1 ), 0.95 (s, 9 ), 0.17 (s, 3 ), 0.16 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 141.6, 140.0, 123.1, 122.2, 80.5, 79.6, 68.2, 55.8, 52.0, 43.1, 38.4, 34.0, 33.2, 25.8, 25.4, 20.6, 18.2, 4.4, 4.6 ppm; RMS (ESI): calcd for C SiNa + [M + Na + ] , found Dimethyl amines 39 and 40: To a stirred solution of epoxide 37 (8.2 mg, mmol) in 2 N (2.0 M solution in TF, 1.0 ml) was added Ti(i-Pr) 4 (34.0 μl, mmol) at room temperature. The TBS TBS resulting mixture was heated to 80 C in a 2 N sealed tube for 50 h and then quenched with 2 N NaC 3 (5 ml, sat. aq.) and stirred for 15 min before it was extracted with EtAc (3 15 ml). The combined organic layers were dried (Na 2 S 4 ), and concentrated in vacuo. Preparative TLC purification (silica gel, C 2 Cl 2 / 10:1) afforded dimethyl amines 39 (6.0 mg, 65%) and 40 (1.5 mg, 16%) as white amorphous solids. 39: R f = 0.30 (silica gel, C 2 Cl 2 / 10:1); IR (film) ν max 3433, 2934, 2858, 1457, 1360, 1297, 1246, 1140, 1054, 832 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.15 (d, J = 1.8 z, 1 ), 5.46 (t, J = 4.2 z, 1 ), 4.04 (d, J = 9.0 z, 1 ), 3.34 (t, J = 9.6 z, 1 ), 2.38 (ddd, J = 13.2, 10.2, 3.0 z, 1 ), 2.28 (s, 6 ), (m, 2 ), 2.05 (ddd, J = 12.0, 9.6, 6.6 z, 1 ), 1.94 (dd, J = 12.6, 3.6 z, 1 ), (m, 4 ),1.78 (td, J = 10.8, 6.6 z, 1 ), (m, 2 ), 0.98 (s, 9 ), 0.19 (s, 3 ), 0.12 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 142.5, 139.6, 121.8, 120.3, 80.9, 79.7, 75.0, 73.8, 62.5, 40.7, 40.1, 38.3, 34.1, 29.2, 26.1, 25.4, 20.7, 18.6, 4.3, 5.0 ppm; RMS (ESI): calcd for C N 3 SiNa + [M + Na + ] , found Diol 38: To a stirred solution of TBS ether 39 (6.0 mg, mmol) in TF (0.3 ml) at room temperature was added TBAF (1.0 M in TF, ml, mmol). The resulting mixture was 2 N 38 stirred for 0.5 h and then quenched with NaC 3 (0.5 ml, sat. aq.) and stirred for 15 min before it was extracted with EtAc (3 5 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. S14

15 Preparative TLC purification (silica gel, ) afforded diol 38 (3.5 mg, 80%) as a white amorphous solid. All physical properties of this compound were identical to those reported for dimethyl amine 38 obtained from epoxy alcohol 36 as previously described. Dimethyl amine 42: To a stirred solution of epoxy alcohol 35 (17.0 mg, mmol) in 2 N (2.0 M solution in TF, 3.0 ml) was added Ti(i-Pr) 4 (101.0 μl, mmol) at room temperature. The resulting mixture was heated to 80 C in a sealed tube for 1 h and then quenched with NaC 3 (5 ml, sat. aq.) and stirred for 15 min before it was 2 N extracted with EtAc (3 15 ml). The combined organic layers were dried 42 (Na 2 S 4 ) and concentrated in vacuo. Preparative TLC purification (silica gel, ) afforded dimethyl amines 42 (14 mg, 70%) as a white amorphous solid. 42: R f = 0.15 (silica gel, ); IR (film) ν max 3407, 2935, 2865, 1456, 1264, 1147, 1069 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.10 (s, 1 ), 5.53 (t, J = 4.2 z, 1 ), 4.45 (d, J = 3.6 z, 1 ), 4.13 (brs, 1 ), 3.54 (dd, J = 10.2, 3.6 z, 1 ), 2.92 (ddd, J = 13.2, 10.2, 3.0 z, 1 ), 2.75 (brs, 1 ), 2.58 (td, J = 11.4, 2.4 z, 1 ), 2.30 (s, 6 ), (m, 2 ), 2.11 (ddd, J = 12.6, 9.6, 7.2 z, 1 ), 2.00 (dd, J = 12.6, 3.0 z, 1 ), (m, 3 ), (m, 2 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 145.3, 140.0, 120.7, 117.5, 81.8, 79.6, 69.5, 66.1, 63.7, 43.5, 39.9, 39.2, 37.9, 34.2, 25.4, 20.6 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Thiocarbonate 44: To a stirred solution of diol 42 (14.0 mg, mmol) in toluene (1.5 ml) was added thiocarbonyldiimidazole (12.8 mg, mmol) at room temperature. The resulting mixture was S 2 N 44 heated to 110 C in a sealed tube for 12 h before it was quenched with NaC 3 (5 ml, sat. aq.) and stirred for 15 min before it was extracted with EtAc (3 15 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 0:1) afforded thiocarbonate 44 (13 mg, 81%) as a white amorphous solid. 44: R f = 0.30 (silica gel, hexanes:etac 0:1); IR (film) ν max 3407, 2935, 2865, 1456, 1264, 1147, 1069 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.03 (s, 1 ), 5.55 (t, J = 3.9 z, 1 ), 5.21 (d, J = 7.2 z, 1 ), 4.73 (dd, S15

16 J = 10.2, 6.6 z, 1 ), 3.28 (ddd, J = 13.2, 10.2, 3.0 z, 1 ), 2.54 (t, J = 9.6 z, 1 ), 2.42 (s, 6 ), 2.19 (quint, J = 4.5 z, 2 ), (m, 2 ), (m, 5 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 170.9, 139.2, 133.4, 128.4, 125.4, 80.4, 80.3, 79.9, 59.1, 52.4, 42.1, 40.9, 37.9, 33.8, 32.5, 25.5, 20.4 ppm; RMS (ESI): calcd for C N 3 SNa + [M + Na + ] , found Triene 45: A solution of thiocarbonate 44 (10.0 mg, 0.03 mmol) in P(Et) 3 (1.0 ml) was heated to 160 C in a sealed tube for 24 h before it was concentrated in vacuo. Preparative TLC purification (silica gel, EtAc: 3:1) afforded triene 45 (3.2 mg, 42%) and 2 N 45 recovered starting material 44 (5.0 mg, 50%) as white amorphous solids. 45: R f = 0.20 (silica gel, EtAc: 3:1); IR (film) ν max 2934, 2862, 1616, 1457, 1275, 1261, 1145, 1043 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.11 (dd, J = 9.6, 2.4 z, 1 ), 5.82 (d, J = 9.6 z, 1 ), 5.79 (s, 1 ), 5.49 (t, J = 3.9 z, 1 ), 3.43 (bd, J = 10.2 z, 1 ), 2.32 (s, 6 ), (m, 2 ), (m, 2 ), 2.02 (t, J = 11.4 z, 2 ), (m, 3 ), (m, 1 ), ppm (m, 2 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 140.8, 140.4, 132.1, 127.2, 122.9, 121.2, 80.4, 80.2, 60.3, 40.6, 38.8, 38.4, 34.1, 31.3, 25.7, 20.8 ppm; RMS (ESI): calcd for C N + [M + + ] , found Diol 46: To a stirred solution of enone 14 2 (8.5 g, 29 mmol) in acetone/ 2 (8:1, 27 ml) were added NM (8.4 g, 72 mmol) followed by s 4 (0.08 M in t-bu, 7.5 ml, 0.59 mmol) at room TBS 46 temperature. The resulting mixture was stirred for 16 h before it was quenched with NaS 3 (50 ml, sat. aq.). The resulting mixture was stirred for 30 min before it was diluted with brine (50 ml) and extracted with EtAc (5 60 ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) afforded diol 46 (6.75 g, 73%) as a clear colorless oil. 46: R f = 0.25 (silica gel, hexanes:etac 3:1); [ α] 25 D = +3.0 (c = 0.1, C 2 Cl 2 ); IR (film) ν max 3446, 2955, 2858, 1715, 1142 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 4.01 (d, J = 11.4 z, 1 ), 3.86 (d, J = 11.4 z, 1 ), 3.58 (t, J = 8.4 z, 1 ), 2.82 (dt, J = 13.8, 6.6 z, 1 ), 2.49 (dddd, J S16

17 = 14.4, 6.6, 4.2, 1.8 z, 1 ), (m, 2 ), (m, 3 ), (m, 1 ), 1.41 (dt, J = 13.6, 4.2 z, 1 ), 1.10 (s, 3 ), 0.89 (s, 9 ), 0.01 ppm (s, 6 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 213.3, 81.5, 80.4, 63.6, 55.2, 43.5, 36.7, 34.4, 31.0, 25.8 (3), 19.7, 18.0, 12.1, 4.5, 4.9 ppm; RMS (ESI): calcd for C SiNa + [M + Na + ] , found Acetonide 47: To a stirred solution of diol 46 (8.0 g, 25 mmol) in acetone (30 ml) were added 2 C() 2 (15 ml, 125 mmol) and p-ts (200 mg, 1.0 mmol) at room temperature. The resulting 47 TBS mixture was stirred for 1 h before it was quenched with NaC 3 (20 ml, sat. aq.) and extracted with Et 2 (3 100 ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 20:1) afforded acetonide 47 (7.95 g, 87%) as a clear colorless oil. 47: R f = 0.3 (silica gel, hexanes:etac 20:1); [ α] 25 D = +8.7 (c = 0.4, C 2 Cl 2 ); IR (film) ν max 2956, 2981, 2862, 1718, 1367, 1142 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 4.07 (d, J = 9.2 z, 1 ), 3.89 (d, J = 9.2 z, 1 ), 3.62 (t, J = 8.5 z, 1 ), 2.57 (dt, J = 13.8, 6.6 z, 1 ), 2.39 (dddd, J = 15.6, 6.5, 4.3, 1.6 z, 1 ), (m, 2 ), 1.84 (dd, J = 12.9, 6.5 z, 1 ), (m, 2 ), (m, 1 ), 1.50 (s, 3 ), (m, 1 ), 1.28 (s, 3 ), 0.90 (s, 3 ), 0.84 (s, 9 ), 0.02 ppm (s, 6 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 208.9, 109.2, 88.2, 80.7, 66.1, 50.9, 44.1, 36.1, 35.6, 30.8, 26.2, 26.0 (3), 25.7, 19.7, 17.9, 11.4, 4.3, 4.5 ppm; RMS (ESI): calcd for C SiNa + [M + Na + ] , found Enoate 48: To a stirred solution of ketone 47 (10.0 g, 27 mmol) in TF (80 ml) was added NaMDS (1.0 M in TF, 33.6 ml, 33.6 mmol) at 0 ο C. The resulting mixture was stirred for 30 min 2 C TBS 48 before PhNTf 2 (10.8 g, 30 mmol) was added. The resulting mixture was stirred for a further 2 h before it was quenched with N 4 Cl (50 ml, sat. aq.) and extracted with Et 2 (3 80 ml). The combined organic layers S17

18 were washed with brine (70 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 20:1) afforded crude enol triflate as a clear colorless oil, which was used for the next step without further purification. To a stirred solution of enol triflate (obtained above, 1.3 g, 2.6 mmol) in DMF (50 ml) were added Pd(PPh 3 ) 4 (150 mg, 0.13 mmol), Et 3 N (1.4 ml, 7.8 mmol), and (20 ml) at room temperature. The resulting mixture was warmed to 70 C and stirred for 3 h under C atmosphere (1 atm) before it was quenched with N 4 Cl (50 ml, sat. aq.). The resulting mixture was extracted with EtAc (3 50 ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 15:1) afforded enoate 48 (766 mg, 72%) as a clear colorless oil. 48: R f = 0.3 (silica gel, hexanes:etac 15:1); [ α] 25 D = +21 (c = 0.2, C 2 Cl 2 ); IR (film) ν max 2953, 2859, 1726, 1368, 1250, 1125, 1029 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.67 (dd, J = 5.4, 2.4 z, 1 ), 4.57 (d, J = 9.0 z, 1 ), 4.16 (d, J = 9.0 z, 1 ), (m, 4 ), 2.14 (dd, J = 6.0, 5.4 z, 1 ), 1.95 (brs, 2 ), (m, 2 ), (m, 2 ), 1.40 (s, 3 ), 1.38 (s, 3 ), 0.87 (s, 9 ), 0.73 (s, 3 ), 0.02 ppm (s, 6 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 168.1, 139.8, 133.9, 106.6, 83.0, 81.3, 66.5, 51.6, 49.7, 43.9, 39.0, 30.1, 27.6, 25.8, 25.7 (3), 21.2, 18.0, 12.5, 4.3, 4.9 ppm; RMS (ESI): calcd for C SiNa + [M + Na + ] , found Alcohol 49: To a stirred solution of TBS ether 48 (7.2 g, 17.5 mmol) in TF (30 ml) was added TBAF (1.0 M in TF, 21 ml, 21 mmol) at room temperature. The resulting mixture was stirred for 16 h 2 C 49 before it was concentrated in vacuo. The resulting residue was diluted with N 4 Cl (30 ml, sat. aq.) and extracted with Et 2 (3 50 ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 3:1) afforded alcohol 49 (4.52 g, 87%) as a clear colorless oil. 49: R f = 0.25 (silica gel, hexanes:etac 2:1); [ α] 25 D = +78 (c = 0.1, CCl 3 ); IR (film) ν max 3447, 2989, 2870, 1724, 1436, 1270, 1143, 1022 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.69 (dd, J = 6.0, 2.4 z, 1 ), 4.57 (d, J = 8.4 z, 1 ), 4.15 (d, J = 8.4 z, 1 ), 3.79 (t, J = 9.0 z, 1 ), 3.72 (s, 3 ), 2.23 (dd, J = 18.6, 6.0 z, 1 ), 2.04 (m, 2 ), 1.92 (dd, J = S18

19 11.4, 7.8 z, 1 ), 1.82 (m, 2 ), 1.53 (m, 1 ), 1.40 (s, 3 ), 1.38 (s, 3 ), 0.76 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 168.1, 139.7, 133.8, 108.7, 82.8, 81.2, 66.5, 51.7, 50.0, 43.5, 38.6, 29.8, 27.6, 26.0, 21.0, 12.2 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Ketone 50: To a stirred solution of alcohol 49 (5.2 g, 17.5 mmol) in C 2 Cl 2 (40 ml) at room temperature were added Dess Martin periodinane (15.75 g, 35.0 mmol) and NaC 3 (7.4 g, C 50 mmol). The resulting mixture was stirred for 3 h before it was diluted with NaC 3 (15 ml, sat. aq.), water (30 ml), and C 2 Cl 2 (100 ml). The layers were separated and the aqueous layer was extracted with Et 2 (3 100 ml). The combined organic layers were washed with brine (70 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 4:1) afforded ketone 50 (4.05 g, 81%) as a white foam. 50: R f = 0.28 (silica gel, hexanes:etac 4:1); [α] 25 D = +57 (c = 0.15, CCl 3 ); IR (film) ν max 2985, 1738, 1632, 1435, 1372, 1238, 1044 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 6.70 (t, J = 4.2 z, 1 ), 4.63 (d, J = 9.0 z, 1 ), 4.17 (d, J = 9.0 z, 1 ), 3.74 (s, 3 ), 2.55 (m, 1 ), 2.24 (m, 4 ), 2.13 (dd, J = 13.2, 4.8 z, 1 ), 1.98 (m, 1 ), 1.47 (s, 3 ), 1.42 (s, 3 ), 0.91 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 219.2, 167.6, 138.5, 134.3, 109.0, 82.8, 66.7, 51.8, 50.6, 47.7, 35.6, 33.9, 27.6, 25.8, 20.5, 15.8 ppm; RMS (ESI): calcd for C Na + [M + Na + ] , found Alkenyl Isoquinoline 51: To a stirred solution of ketone 50 (1.8 g, 6.1 mmol) in TF (60 ml) was added NaMDS (1.0 M in TF, 9.3 ml, 9.3 mmol) at 78 C. The resulting mixture was stirred for 30 min before PhNTf 2 (2.62 g, 7.3 mmol) was added. The resulting 2 C N mixture was warmed to 0 ο C and stirred for 2 h before it was quenched with N 4 Cl (20 ml, sat. aq.) and extracted with EtAc ( ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 5:1) afforded enol triflate of 50 (2.9 g) as a colorless oil. To a stirred solution of the enol triflate (obtained above, 2.9 g, ca 6.1 mmol) in TF (degassed, 60 ml) were added Pd(PPh 3 ) 4 (704 mg, 0.61 mmol), K 2 C 3 (3.37 g, 24.4 mmol), and isoquinoline boronic S19

20 ester 11 3 (2.0 g, 7.93 mmol) at room temperature. The resulting mixture was warmed to 80 C and stirred for 3 h before it was concentrated in vacuo. The resulting residue was diluted with N 4 Cl (50 ml, sat. aq.) and EtAc (100 ml), the layers were separated, and the aqueous layer was extracted with EtAc (3 50 ml). The combined organic layers were washed with brine (50 ml), dried (MgS 4 ), and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 2:1) afforded alkenyl isoquinoline 51 (1.54 g, 62% over two steps) as a yellow oil. 51: R f = 0.35 (silica gel, hexanes:etac 1:1); [ α] 25 D = +2.0 (c = 0.2, C 2 Cl 2 ); IR (film) ν max 2924, 2870, 1724, 1458, 1379, 1274, 1137, 1079 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 9.22 (s, 1 ), 8.49 (d, J = 6.0 z, 1 ), 7.87 (s, 1 ), 7.78 (d, J = 9.0 z, 1 ), 7.76 (dd, J = 8.4, 1.8 z, 1 ), 7.62 (d, J = 5.4 z, 1 ), 6.79 (dd, J = 6.0, 2.4 z, 1 ), 6.24 (dd, J = 3.0, 1.8 z, 1 ), 4.69 (d, J = 9.0 z, 1 ), 4.35 (d, J = 9.0 z, 1 ), 3.77 (s, 3 ), 2.64 (dd, J = 18.0, 5.4 z, 1 ), (m, 1 ), 2.57 (dd, J = 10.8, 1.8 z, 1 ), (m, 2 ), 1.50 (s, 3 ), 1.46 (s, 3 ), 1.16 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 167.9, 152.6, 149.6, 143.1, 139.1, 135.1, 134.9, 134.8, 129.9, 128.8, 128.7, 126.6, 124.0, 120.1, 108.7, 82.4, 67.1, 55.9, 51.7, 47.9, 38.2, 30.8, 27.6, 26.1, 18.8 ppm; RMS (ESI): calcd for C N 4 Na + [M + Na + ] , found Isoquinoline 52: thod A: To a stirred solution of alkenyl isoquinoline 51 (38 mg, mmol) in Et (10.0 ml) was added Wilkinson s catalyst (2.6 mg, mmol) at room temperature. The resulting mixture was stirred under 2 (1 atm) for 12 h before it was filtered and washed with EtAc (3 50 ml). The combined organic layers were concentrated in vacuo. LCMS analysis of the crude reaction mixture indicated no desired product. thod B: To a stirred solution of alkenyl isoquinoline 51 (45 mg, 0.11 mmol) in TF (10.0 ml) was added Cy 2 B [1 M in TF (prepared from hydroboration of cyclohexene with B 3 S 2 ), 0.28 ml, 0.28 mmol] at room temperature. The resulting mixture was stirred for 10 h before it was quenched with Ac (0.13 ml, 2.2 mmol). The resulting mixture was stirred 2 h at room temperature before it was diluted with water (10 ml) and extracted with EtAc (3 50 ml). The combined organic layers S20

21 N were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column 2 C 52 chromatography (silica gel, hexanes:etac 1:1) afforded isoquinoline 52 (6.3 mg, 14% yield) as a white solid. 52: R f = 0.20 (silica gel, hexanes:etac 1:1); [ α] 25 D = +5.0 (c = 0.4, CCl 3 ); m.p. = C (C 2 Cl 2 /hexanes); IR (film) ν max 2916, 2189, 2160, 1980, 1723, 1141 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 9.24 (s, 1 ), (d, J = 5.4 z, 1 ), 7.78 (s, 1 ), 7.77 (s, 1 ), 7.63 (d, J = 5.4 z, 1 ), 7.57 (d, J = 1.2 z, 1 ), 6.68 (dd, J = 5.4, 1.8 z, 1 ), 4.63 (d, J = 8.4 z, 1 ), 4.21 (d, J = 8.4 z, 1 ), 3.75 (s, 3 ), 3.12 (dd, J = 11.4, 7.8 z, 1 ), 2.35 (t, J = 11.4 z, 1 ), 2.30 (dd, J = 11.4, 7.8 z, 1 ), (m, 2 ), 2.03 (d, J = 5.4 z, 1 ), 2.02 (d, J = 4.2 z, 1 ), 1.87 (s, 1 ), 1.49 (s, 3 ), 1.43 (s, 3 ), 0.55 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 168.1, 152.3, 142.7, (2), 134.7, 134.2, 131.7, 128.6, 126.4, 126.0, 120.1, 108.8, 83.0, 66.5, 56.7, 54.9, 51.7, 45.0, 39.8, 27.7, 26.1, 25.9, 22.4, 14.3 ppm; RMS (ESI): calcd for C N 4 Na + [M + Na + ] , found thod C: To a stirred solution of alkenyl isoquinoline 51 (51 mg, 0.13 mmol) in TF (10.0 ml) was added B 3 S 2 (0.02 ml, 0.20 mmol) at room temperature. The resulting mixture was stirred for 5 h before it was quenched with Ac (0.015 ml, 0.26 mmol). The resulting mixture was stirred for 2 h before it was diluted with water (10 ml) and extracted with EtAc (3 50 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 1:1) afforded isoquinoline 52 (10 mg, 20% yield) as a white amorphous solid. thod D: To a stirred solution of alkenyl isoquinoline 51 (40 mg, mmol) in (10.0 ml) was added Pd-C (10% w/w, 20 mg, mmol) at room temperature. The resulting mixture was stirred under 2 (1 atm) for 4 h before it was filtered and washed with EtAc (3 50 ml). The combined organic layers were concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 1: 1) afforded isoquinoline 52 (28 mg, 70% yield) as a white amorphous solid. thod E: To a stirred solution of alkenyl isoquinoline 51 (67 mg, 0.16 mmol) and K 2 CN=NC 2 K (80 mg, 0.48 mmol) in TF/ 2 (4:1, 10.0 ml) was added Ac (0.18 ml, 3.2 mmol) via syringe S21

22 pump at room temperature. The resulting mixture was stirred for 6 h before it was quenched with N 4 Cl (5 ml, sat. aq.) and filtered through a short silica gel column. The residue was diluted with water (10 ml) and extracted with EtAc (3 50 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 1:1) afforded isoquinoline 52 (16.8 mg, 25% yield) as a white amorphous solid. Alcohol 53: To a stirred solution of the methyl ester 52 (250 mg, 0.61 mmol) in toluene (28 ml) was added DIBAL (1.0 M in toluene, 1.8 ml, 1.8 mmol) at 78 C. The resulting mixture was stirred for N 3 h before it was quenched with (2 ml) and NaC 3 (15 ml, sat. 53 aq.), and allowed to warm to room temperature and stirred for 30 min. The resulting mixture was extracted with EtAc (4 50 ml). The combined organic layers were dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 1:3) afforded alcohol 53 (166 mg, 71%) as a white amorphous solid. 53: R f = 0.25 (silica gel, hexanes:etac 1:2); [ α] 25 D = (c = 0.3, C 2 Cl 2 ); IR (film) ν max 3254, 2989, 2870, 1598, 1458, 1393, 1379, 1360, 1293, 1077 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 1 NMR (600 Mz, CDCl 3 ): δ = 9.30 (s, 1 ), 8.50 (d, J = 6.0 z, 1 ), 7.78 (s, 1 ), 7.77 (d, J = 9.0 z, 1 ), 7.64 (d, J = 5.4 z, 1 ), 7.57 (dd, J = 8.4, 1.8 z, 1 ), 5.89 (dd, J = 5.4, 1.8 z, 1 ), 4.26 (d, J = 9.6 z, 1 ), 4.24 (bs, 2 ), 3.96 (d, J = 9.6 z, 1 ), 3.15 (dd, J = 11.4, 9.0 z, 1 ), 2.45 (dd, J = 12.0, 8.4 z, 1 ), (m, 2 ), (m, 2 ), 1.99 (td, J = 12.0, 4.8 z, 1 ), 1.93 (dd, J = 16.8, 5.4 z, 1 ), 1.56 (s, 3 ), 1.48 (s, 3 ), 0.51 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 151.9, 141.9, 139.8, 139.7, 134.9, 132.2, 129.1, 128.6, 126.4, 126.0, 120.4, 108.1, 84.5, 67.4, 64.1, 56.9, 54.3, 45.1, 39.7, 26.8, 26.1, 26.0, 22.3, 14.5 ppm; RMS (ESI): calcd for C N 3 Na + [M + Na + ] , found Enal 54: To a stirred solution of alcohol 53 (250 mg, 0.66 mmol) in C 2 Cl 2 (6.0 ml) were added NaC 3 (276 mg, 3.3 mmol) and DMP (410 mg, 0.99 mmol) at room temperature. The reaction mixture was stirred for 30 min before it was quenched with NaC 3 (10 ml, sat. aq.) and Na 2 S 2 3 (10 ml, sat. aq.), and then extracted with C 2 Cl 2 (4 30 ml). The combined organic layers were dried S22

23 N (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica 54 gel, hexanes:etac 1:3) afforded enal 54 (200 mg, 80%) as a white amorphous solid. 54: R f = 0.55 (silica gel, hexanes:etac 1:2); [ α] 25 D = (c = 0.1, C 2 Cl 2 ); IR (film) ν max 2937, 1696, 1592, 1479, 1369, 1241, 1202, 1150 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 9.66 (s, 1 ), 9.25 (s, 1 ), 8.52 (d, J = 4.8 z, 1 ), 7.80 (s, 1 ), 7.79 (d, J = 8.4 z, 1 ), 7.65 (d, J = 5.4 z, 1 ), 7.58 (dd, J = 9.0, 1.8 z, 1 ), 6.92 (dd, J = 6.0, 2.4 z, 1 ), 4.28 (d, J = 8.4 z, 1 ), 4.23 (d, J = 9.0 z, 1 ), 3.16 (dd, J = 10.8, 7.8 z, 1 ), 2.49 (br d, J = 18.6 z, 1 ), (m, 2 ), (m, 2 ), 2.18 (ddd, J = 13.2, 9.6, 4.2 z, 1 ), 2.03 (ddd, J = 15.6, 11.4, 4.2 z, 1 ), 1.60 (s, 3 ), 1.53 (s, 3 ), 0.53 ppm (s, 3 ); 13 C NMR (150 Mz, CDCl 3 ): δ = 192.4, 152.3, 150.7, 142.8, 141.2, 138.9, 134.8, 131.7, 126.5, 126.2, 120.1, 109.3, 81.7, 66.7, 56.5, 55.1, 45.5, 40.4, 27.6, 26.1, 26.0, 22.1, 14.3 ppm; RMS (ESI): calcd for C N 3 Na + [M + Na + ] , found Dithiane diol 55: To a stirred solution of aldehyde 54 (200 mg, 0.53 mmol) in C 2 Cl 2 (11 ml) were added S(C 2 ) 3 S (0.42 ml, 3.2 mmol) and BF 3 Et 2 (0.72 ml, 5.3 mmol) at 78 C. The reaction N mixture was warmed to 40 C and stirred for 3 h before it was quenched with (2 ml) and NaC 3 (15 ml, sat. aq.), and then stirred S S vigorously for 30 min at room temperature. The resulting mixture was extracted with EtAc (4 50 ml) and the combined organic layers were 55 dried (Na 2 S 4 ) and concentrated in vacuo. Flash column chromatography (silica gel, hexanes:etac 0:1) afforded dithiane diol 55 (92 mg, 41%) as a white amorphous solid. 55: R f = 0.33 (silica gel, hexanes:etac 0:1); [ α] 25 D = (c = 0.05, C 2 Cl 2 ); IR (film) ν max 3356, 2958, 2896, 1595, 1505, 1273, 1034 cm 1 ; 1 NMR (600 Mz, CDCl 3 ): δ = 9.24 (s, 1 ), 8.50 (d, J = 6.0 z, 1 ), 7.78 (d, J = 8.4 z, 1 ), 7.77 (s, 1 ), 7.65 (d, J = 5.4 z, 1 ), 7.56 (d, J = 8.4 z, 1 ), 6.27 (dd, J = 5.4, 2.4 z, 1 ), 4.90 (s, 1 ), 3.90 (dd, J = 12.0, 4.8 z, 1 ), 3.84 (dd, J = 11.4, 8.4 z, 1 ), 3.09 (t, J = 9.6 z, 1 ), 2.99 (m, 2 ), 2.76 (dd, J = 8.4, 4.8 z, 2 ), 2.36 (d, J = 18.0 z, 1 ), 2.26 (m, 1 ), 2.15 (m, 4 ), 2.05 (m, 1 ), 2.00 (dd, J = 17.4, 6.0 z, 1 ), 1.85 (m, 1 ), 0.58 ppm (s, 3 ); 13 C NMR (150 S23