Supporting Information

Transcription

1 Supporting Information Wiley-VCH Weinheim, Germany

2 Supporting Information First Asymmetric Total Synthesis of (+)-Cytotrienin A, an Ansamycin-type Anticancer Drug Yujiro Hayashi,* Mitsuru Shoji, Hayato Ishikawa, Junichiro Yamaguchi, Tomohiro Tamura, Hiroki Imai, Yosuke Nishigaya, Kenichi Takabe, Hideaki Kakeya, Hiroyuki sada Department of Industrial Chemistry, Faculty of Engineering, Tokyo University of Science, Kagurazaka, Shinjuku-ku, Tokyo , Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto-city, Kyoto , Antibiotics Laboratory, RIKEN Discovery Research Institute, Wako, Saitama , Japan General Remarks: All reactions were carried out under argon atmosphere and monitored by thin-layer chromatography using rck 60 F254 precoated silica gel plates (0.25 mm thickness). Specific optical rotations were measured using a J=ASC P-1020 polarimeter. FT-IR spectra were recorded on a JASC FT/IR-410 spectrometer. 1 H and 13C NMR spectra were recorded on a Brucker AM-400 (400 MHz for 1 H NMR, 100 MHz for 13 C NMR) and Brucker AV-600 (600 MHz for 1 H NMR, 150 MHz for 13 C NMR) instrument. Data for 1 H NMR are reported as chemical shift (δ ppm), multiplicity (s = singlet, d = doublet, t = triplet, dd = doubldoublet, dt = doubltriplet, m = multiplet, br = broad), coupling constant (Hz), integration, and assignment. Data for 13 C NMR are reported as chemical shift. High-resolution mass spectral analyses (HRMS) were carried out using Bruker ESI-TF MS. Preparative thin layer chromatography was performed using Wakogel B-5F purchased from Wako Pure Chemical Industries, Tokyo, Japan. Flash chromatography was performed using silica gel 60N of Kanto Chemical Co. Int., Tokyo, Japan. HPLC analysis was performed on a HITACHI Elite LaChrom Series HPLC, UV detection monitered at appropriate wavelength respectively, using Chiralcel Chiralpak IC (0.46 cm 25 cm) or Chiralpak AS-H (0.46 cm 25 cm). - S1 -

3 Experimental Data (1R, 2R)-1-(Furan-2-yl)-2-methylpropane-1,3-diol (2) 8 10 mol% H + H N H 33 neat, 4 o C, 48 h C 2 H then NaBH 4, H 0 o C, 1 h H H 2 H H + S1 S2 H 77%, 96% ee anti : syn = 6.2 : 1 To a solution of furfural (0.91 ml, 10.4 mmol) and 4-acyloxyproline 33 (297 mg, 1.04 mmol) was added propanal (2.25 ml, 31.2 mmol) at 4 C under argon atmosphere. The reaction mixture was stirred for 48 h at that temperature followed by an addition of H (10 ml) and NaBH4 (1.2 g, 31.2 mmol). The reaction mixture was stirred for additional 1 h at 0 C before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (5 times). The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration followed by an azeotropic removal with H (3 times). Compound S1 and trace amount of furfuryl alcohol (S2) were removed by Kugel-Rohr (115~120 C, 0~1 Torr) and the residue was directly passed through silica gel pad eluted with ethyl acetate to afford 2 (1.25 g, 77%, anti : syn = 6.2 : 1, 96% ee) as a pale yellow oil. This compound was identified with published data 1). For HPLC analysis; The product 2 was converted to the corresponding mono benzoylester with benzoylchloride, pyridine and enantiomeric excess was determined by HPLC with a Chiralpak AS H column (1 : 100 = 2-propanol : hexane), 0.5 ml/min ; major enantiomer tr = 45.7 min, minor enantiomer tr = 52.0 min. (2R, 4R, 5R)-4-(Furan-2-yl)-2-(4-methoxyphenyl)-5-methyl-1,3-dioxane (3) PMP H H PMPC() 2 PPTS benzene 80 C, 1 h 2 3 single isomer 64%, >99% ee - S2 -

4 To a solution of 2 (1.22 g, anti : syn = 6.2 : 1, 7.81 mmol) in benzene (16 ml) were added p-anisaldehyde dimethylacetal (1.6 ml, 9.37 mmol) and pyridinium p-toluene sulfonate (PPTS, 98 mg, 0.39 mmol) at room temperature under argon atmosphere. The resulting mixture was stirred for 1 h at 80 C before being quenched with saturated aqueous NaHC3. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl (3 times). The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by recrystallization from 33% ethyl acetate in n-hexane to afford acetal 3 (1.36 g, 64%, >99% ee) as white needles. 1H NMR (400 MHz, CDCl3) δ 0.74 (3H, d, J = 6.7 Hz), (1H, m), 3.61 (1H, t, J = 11.4 Hz), 3.78 (3H, s), 4.26 (1H, dd, J = 11.4, 4.7 Hz), 4.49 (1H, d, J = 10.4 Hz), 5.59 (1H, s), 6.36 (1H, dd, J = 3.2, 1.0 Hz), 6.39 (1H, br d, J = 3.2 Hz), 6.87 (2H, br d, J = 8.7 Hz), 7.41 (1H, d, J = 1.0 Hz), 7.45 (2H, br d, J = 8.7 Hz); 13C NMR (100 MHz, CDCl3) δ 12.4, 32.8, 55.3, 73.2, 78.7, 101.6, 108.5, 110.1, 113.6, 127.6, 130.7, 142.4, 152.5, 160.0; IR (KBr) ν 2958, 2927, 2845, 1618, 1587, 1520, 1455, 1425, 1392, 1354, 1308, 1254, 1174, 1074, 1036, 822, 748, 652, 606 cm 1; HRMS (ESI) [M+Na] + calculated for [C16H18Na4] + : , found: ; [α]d (c 0.99, H). mp C Enantiomeric excess was determined by HPLC with a Chiralpak IC column (1 : 30 = 2-propanol : hexane), 1.3 ml/min ; major enantiomer tr = 8.5 min, minor enantiomer tr = 10.0 min. (2R, 3R)-3-(Furan-2-yl)-3-(4-methoxybenzyloxy)-2-methylpropane-1-ol (4) PMP DIBAL-H PMB Et 2!78 ~!10 o C h 80 % 4 (borsm : 92%) H To a solution of p methoxyphenyl acetal 3 (24 g, 87.5 mmol) in Et2 (875 ml) was added diisobutylaluminium hydride (DIBAL-H, 268 ml, 0.98 M hexane solution, S3 -

5 mmol) at 78 C under argon atmosphere. The reaction temperature was raised to 10 C over 128 h before being quenched with an addition of saturated aqueous Rochelle salt. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 10/1) to afford alcohol 4 (19.3 g, 80%) as a colorless oil and recovered 5 (3.1 g, 13%). 1H NMR (400 MHz, CDCl3) δ 0.71 (3H, d, J = 7.0 Hz), (1H, m), 2.87 (1H, br s), 3.59 (1H, dd, J = 10.7, 7.0 Hz) (1H, m), 3.80 (3H, s), 4.21 (1H, d, J = 11.5 Hz), 4.22 (1H, d, J = 9.0 Hz), 4.45 (1H, d, J = 11.5 Hz), 6.32 (1H, br d, J = 3.1 Hz), 6.38 (1H, dd, J = 3.1, 1.3 Hz), 6.87 (2H, br d, J = 8.6 Hz), 7.21 (2H, br d, J = 8.6 Hz), 7.44 (1H, d, J = 1.3 Hz); 13C NMR (100 MHz, CDCl3) δ 13.4, 39.4, 55.1, 66.7, 70.0, 78.5, 109.0, 109.9, 113.8, 129.4, 129.6, 142.3, 153.0, 159.2; IR (neat) ν 3448, 2962, 1612, 1514, 1466, 1302, 1250, 1174, 1151, 1035, 1010, 822, 741 cm 1; HRMS (ESI) [M+Na] + calculated for [C16H20Na4] + : , found: ; [α]d (c 0.77, H). (2S, 3R)-3-(Furan-2-yl)-3-(4-methoxybenzyloxy)-2-methylpropanal (5) PMB H S 3 pyr DMS, Et 3 N CH 2 Cl 2 0 C, 45 min PMB 4 quant. 5 H To a solution of alcohol 4 (4.7 g, 17 mmol) in CH2Cl2 (17 ml) were added triethylamine (11.8 ml, 85 mmol), dimethylsulfoxide (DMS, 17 ml, 1.0 M) and S3 pyridine complex (7.8 g, 51 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 45 min at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 10/1) to afford aldehyde 5 (4.7 g, quantitative yield) as a pale yellow oil. - S4 -

6 1H NMR (400 MHz, CDCl3) δ 0.86 (3H, d, J = 7.1 Hz), (1H, m), 3.78 (3H, s), 4.21 (1H, d, J = 11.5 Hz), 4.45 (1H, d, J = 11.5 Hz), 4.48 (1H, d, J = 9.1 Hz), (2H, m), 6.84 (2H, br d, J = 8.5 Hz), 7.16 (2H, br d, J = 8.5 Hz), 7.44 (1H, br s), 9.76 (1H, br d, J = 2.3 Hz); 13C NMR (100 MHz, CDCl3) δ 10.7, 49.9, 55.2, 70.1, 74.5, 109.8, 110.1, 113.8, 129.5, 129.6, 143.0, 151.7, 159.3, 203.3; IR (neat) ν 2993, 2851, 1729, 1613, 1514, 1457, 1249, 1063, 819, 739 cm 1; (1R, 2R, 3R, Z)-1-(Furan-2-yl)-1-(4-methoxybenzyloxy)-2,4-dimethyl-6- (triisopropylsiloxy)hex-4-en-3-ol (6) PMB H 5 I TIPS 34 t BuLi, 2 Zn THF,!78 o C~ 0 o C 79% PMB H TIPS 6 To a solution of 34 (1.46 g, 4.13 mmol) in THF (20 ml) was added t-buli (5.2 ml, 8.27 mmol, 1.59 M solution in pentane) at 78 C under argon atmosphere. The reaction mixture was stirred for 1 h at that temperature followed by an addition of 2Zn (4.13 ml, 4.13 mmol, 1.0 M solution in hexane). After the reaction mixture was stirred for additional 20 min at 0 C and then cooled to 78 C, aldehyde 5 (945 mg, 3.45 mmol) in THF (20 ml) was added dropwise to the reaction mixture. The reaction temperature was raised slowly to 35 C over 3 h before being quenched with ph 7.0 phosphate buffer solution. After the crude material was filtrated through celite pad, the organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 50/1) to afford alcohol 6 (1.36 g, 79%) as a pale yellow oil. 1H NMR (400 MHz, CDCl3) δ 0.83 (3H, d, J = 7.0 Hz), 1.05 (21H, s), 1.73 (3H, s), 2.14 (1H, dq, J = 3.2, 7.0 Hz), 2.75 (1H, br d, J = 3.1 Hz), 3.80 (3H, s), (2H, m), S5 -

7 (1H, d, J = 11.4 Hz), 4.37 (1H, d, J = 7.0Hz), 4.48 (1H, d, J = 11.4 Hz), 4.74 (1H, br s), 5.42 (1H, dt, J = 1.0, 5.6 Hz), 6.32 (1H, d, J = 3.1 Hz), 6.37 (1H, dd, J = 1.8, 3.1 Hz), 6.86 (2H, d, J = 8.5 Hz), 7.22 (2H, d, J = 8.5 Hz), 7.42 (1H, br d, J = 1.8 Hz); 13C NMR (100 MHz, CDCl3) δ 10.7, 12.0, 18.0, 20.5, 42.2, 55.2, 59.5, 70.7, 71.2, 76.7, 108.7, 110.0, 113.8, 127.0, 129.5, 130.1, 138.4, 142.2, 153.6, 159.3; IR (neat) ν 2942, 2866, 1614, 1514, 1458, 1386, 1249, 1173, 1056, 1011, 882, 821, 738, 683, 432 cm 1; HRMS (ESI) [M+Na] + calculated for [C29H46Na5Si] + : , found: ; [α]d (c 1.54, H). (1R, 2S, 3R, Z)-1-(Furan-2-yl)-1-(4-methoxybenzyloxy)-2,4-dimethyl-3,6-bi s (triisopropylsiloxy)hex-4-ene (7) PMB H TIPSTf 2,6-lutidine PMB TIPS TIPS CH 2 Cl 2 0 o C, 23 h 6 99% 7 TIPS To a solution of alcohol 6 (5.0 g, 9.95 mmol) in CH2Cl2 (99 ml) were added 2,6-lutidine (1.97 ml, 16.9 mmol) and triisopropylsilyl trifluoromethaneslfonate (TIPSTf, 4.0 ml, 14.9 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 23 h at that temperature before being quenched with saturated aqueous NaHC3. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 100/1) to afford 7 (6.43 g, 99%) as a pale red oil. 1H NMR (400 MHz, CDCl3) δ 0.85 (3H, d, J = 6.8 Hz), 1.05 (42H, s), 1.75 (3H, s), (1H, m), 3.80 (3H, s), 4.16 (1H, br ddd, J = 12.8, 4.4, 1.6 Hz), 4.21 (1H, d, J = 11.2 Hz), 4.25 (1H, d, J = 7.6 Hz), (2H, m), 4.80 (1H, d, J = 4.4 Hz), 5.30 (1H, br t, J = 6.0 Hz), 6.29 (1H, br d, J = 3.2 Hz), 6.35 (1H, br dd, J = 3.2, 1.6 Hz), 6.83 (2H, d, J = 8.8 Hz), 7.18 (2H, d, J = 8.4 Hz), 7.39 (1H, br d, J = 1.2 Hz); 13C NMR (100 MHz, CDCl3) δ 11.1, 12.0, 12.9, 18.0, 18.2, 18.2, 19.3, 44.4, 55.2, 59.7, 69.9, 70.9, 75.6, 109.2, 109.9, 113.6, 127.3, 129.2, 130.7, 138.3, 142.0, 153.9, 159.0; - S6 -

8 IR (neat) ν 2942, 2866, 2360, 1614, 1514, 1463, 1384, 1249, 1173, 1105, 1060, 1011, 919, 883, 821, 737, 682, 599, 421 cm 1; HRMS (ESI) [M+Na] + calculated for [C38H66Na5Si2] + : , found: ; [α]d (c 1.71, H). (2E, 5R, 6S, 7R, 8Z)-5-(4-thoxybenzyloxy)-6,8-dimethyl-7,10-bis (triisopropylsiloxy)deca-2,8-diene-1,4-diol (8) PMB TIPS 1) 2, h! Rose bengal EtCN, "78 C, 8 h ; 2 S "20 C, 15 h 2) DABC EtCN, "20 C, 2 h TIPS PMB NaBH 4 CeCl 3 7H 2 EtH, 0 C 20 min TIPS PMB TIPS 7 S3 81% (2 steps) TIPS H TIPS H 8 H A solution of 7 (500 mg, 0.76 mmol) in propionitrile (EtCN, 15.2 ml) was irradiated for 8 h at 78 C with a fluorescent lamp (22 W) in the presence of Rosebengal (38.7 mg, mmol) under oxygen atmosphere followed by an adittion of 2S (3.3 ml, 45.6 mmol) under argon atmosphere. After the reaction temperature was raised to 20 C over 15 h, 1,4-diazabicyclo[2.2.2]octane (DABC, 85.2 mg, 0.76 mmol) in propionitrile (EtCN, 2 ml) was added to the solution of aldehyde mixture. The resulting mixture was stirred for 2 h at that temperature before being quenched with saturated aqueous NH4Cl followed by a filtration through celite pad. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NH4Cl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 10/1) to afford unstable aldehyde S3 (493 mg) as a pale yellow oil. To the solution of aldehyde S3 (493 mg) and CeCl3 7H2 (812 mg, 2.18 mmol) in EtH (18.3 ml) was added NaBH4 (82.5 mg, 2.18 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 20 min at that temperature before being quenched with saturated aqueous NH4Cl. The organic materials were extracted with chloroform (2 times) and ethyl acetate (2 times). The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 5/1) to afford diol 8 (417 mg, 81%, 2 steps) as a pale orange oil. - S7 -

9 1H NMR (400 MHz, CDCl3) δ 1.04 (21H, br s), 1.07 (21H, br s), 1.18 (3H, d, J = 6.8 Hz), 1.73 (3H, s), (1H, m), 2.63 (1H, d, J = 7.2 Hz), 3.25 (1H, t, J = 3.2 Hz), 3.80 (3H, s), (4H, m), 4.33 (1H, dd, J = 12.8, 7.2 Hz), 4.41 (1H, d, J = 11.2 Hz), 4.48 (1H, d, J = 8.4 Hz), 4.59 (1H, d, J = 8.4 Hz), 5.36 (1H, dd, J = 12.0, 6.0 Hz), 5.66 (1H, dd, J = 15.6, 7.2 Hz), 5.89 (1H, dt, J = 15.6, 10.4 Hz), 6.86 (2H, d, J = 8.4 Hz), 7.22 (2H, d, J = 8.4 Hz) (H 1, undetected); 13C NMR (100 MHz, CDCl3) δ 12.0, 12.7, 13.1, 18.0, 18.2, 18.3, 40.6, 55.3, 59.7, 63.0, 70.8, 72.0, 72.6, 81.1, 113.8, 128.4, 129.4, 130.3, 130.4, 132.7, 137.3, 159.3; IR (neat) ν 3419, 2943, 2866, 1614, 1515, 1464, 1384, 1302, 1250, 1173, 1056, 1013, 883, 822, 777, 682, 595 cm 1; HRMS (ESI) [M+Na] + calculated for [C38H70Na6Si2] + : , found: ; [α]d (c 0.46, H). (2E, 5R, 6S, 7R, 8Z)-5-(4-thoxybenzyloxy)-6,8-dimethyl-7,10-bis triisopropylsiloxy-1-triphenylmethoxy-deca-2,8-dien-4-ol (9) TIPS PMB TIPS TIPS TrCl, Et 3 N CH 2 Cl 2 TIPS 23 C, 3 h H PMB 93% H H 8 9 Tr To a solution of diol 8 (7.9 g, 11.6 mmol) in CH2Cl2 (23 ml) were added Et3N (6.5 ml, 46.4 mmol) and triphenylmethyl chloride (TrCl, 6.5 g, 23.2 mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 3 h at that temperature before being quenched with excess H followed by an addition of ph 7.0 phosphate buffer solution. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 30/1) to afford 9 (9.3 g, 93%) as a pale yellow oil. 1H NMR (400 MHz, CDCl3) δ 0.99 (21H, s), 1.07 (21H, br s), 1.18 (3H, d, J = 6.8 Hz), 1.72 (3H, s), ,13 (1H, m), 2.54 (1H, d, J = 7.8 Hz), 3.29 (1H, dd, J = 4.0, 2.8 Hz), 3.61 (2H, br d, J = 2.4 Hz), 3.76 (3H, s), (2H, m), 4.32 (1H, dd, J = 12.8, 7.8 Hz), - S8 -

10 4.51 (1H, dd, J = 18.6, 7.8 Hz), 4.53 (1H, d, J = 7.2 Hz), 4.59 (1H, d, J = 10.8 Hz), 5.33 (1H, br t, J = 6.4 Hz), 5.84 (1H, d, J = 3.2 Hz), 5.86 (1H, d, J = 4.0 Hz), 6.80 (2H, d, J = 8.4 Hz), (11H, m), 7.45 (6H, d, J = 7.2 Hz); 13C NMR (100 MHz, CDCl3) δ 12.0, 12.8, 13.1, 18.0, 18.2, 18.3, 41.0, 55.2, 59.6, 64.2, 70.9, 72.0, 73.0, 81.4, 86.8, 113.8, 126.9, 127.7, 128.1, 128.6, 129.3, 130.3, 133.0, 137.6, 144.2, 159.3; IR (neat) ν 2943, 2866, 1614, 1514, 1458, 1386, 1249, 1173, 1056, 882, 822, 764, 704, 681 cm 1; HRMS (ESI) [M+Na] + calculated for [C57H84Na6Si2] + : , found: ; [α]d (c 0.92, H). (2E, 5R, 6S, 7R, 8Z)-Formic acid 2-(4-methoxybenzyloxy)-3,5-dimethyl-4,7-bis triisopropylsiloxy-1-(3- triphenylmethoxypropenyl)-hept-5-enyl ester (10) TIPS PMB TIPS DMAP N N N CH TIPS TIPS Tr CH 2 Cl 2, 23 C PMB 4 h H quant. CH 9 10 Tr To a solution of alcohol 9 (3.7 g, 4.02 mmol) and 4-dimethylaminopyridine (DMAP, 147 mg, 1.21 mmol) in CH2Cl2 (8.0 ml) was added N-formylbenzotriazole S2) (1.7 g, 9.25 mmol, purity : 80%) at 23 C under argon atmosphere. The reaction mixture was stirred for 4 h at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 30/1) to afford 10 (3.8 g, quantitative yield) as a pale yellow oil. 1H NMR (600 MHz, CDCl3) δ 0.99 (21H, s), 1.06 (21H, br s), 1.12 (3H, d, J = 7.2 Hz), 1.71 (3H, s), 2.04 (1H, dq, J = 12.1, 5.1 Hz), 3.41 (1H, t, J = 4.8 Hz), 3.60 (2H, br d, J = 4.2 Hz), 3.75 (3H, s), 4.18 (1H, br dd, J = 12.8, 4.1 Hz), 4.37 (1H, dd, J = 12.8, 7.9 Hz), 4.59 (2H, s), 4.62 (1H, d, J = 7.2 Hz), 5.35 (1H, br t, J = 6.4 Hz), 5.55 (1H, br t, J = 5.1 Hz), 5.84 (1H, - S9 -

11 dt, J = 15.5, 8.3 Hz), 5.91 (1H, dd, J = 15.7, 6.4 Hz), 6.78 (2H, d, J = 9.0 Hz), (11H, m), 7.43 (6H, d, J = 7.8 Hz), 8.11 (1H, s); 13C NMR (100 MHz, CDCl3) δ 12.0, 12.8, 13.6, 18.0, 18.2, 18.2, 40.1, 55.1, 59.8, 63.8, 71.3, 73.3, 73.9, 81.5, 86.9, 113.7, 126.2, 127.0, 127.8, 128.5, 128.6, 129.3, 130.4, 131.7, 137.4, 144.0, 159.2, 160.3; IR (neat) ν 2943, 2866, 2724, 1727, 1613, 1587, 1514, 1491, 1463, 1449, 1383, 1248, 1173, 1057, 1013, 883, 822, 763, 745, 704, 682, 633, 595, 462 cm 1; HRMS (ESI) [M+Na] + calculated for [C58H84Na7Si2] + : , found: ; [α]d (c 1.51, H). (2Z, 4R, 5S, 6S, 8E)-6-(4-thoxybenzyloxy)-3,5-dimethyl-1,4-bis(triisopropylsiloxy) -10- triphenylmethoxy-deca-2,8-diene ( 11) TIPS PMB TIPS Pd 2 (dba) 3 CHCl 3 TIPS n Bu 3 P, HC 2 NH 4 TIPS 1.4-dioxane, 23 C Tr 67 h PMB CH 76% Tr To a solution of aldehyde 10 (940 mg, 0.99 mmol), formic acid ammonium salt (125 mg, 1.98 mmol) and tributylphosphine (0.39 ml, 1.58 mmol) in degassed 1,4-dioxane (9.9 ml) was added Pd2(dba)3 CHCl3 (205 mg, 0.20 mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 67 h at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 40/1) to afford 11 (683 mg, 76%) as a pale yellow oil. 1H NMR (400 MHz, CDCl3) δ (45H, m), 1.71 (3H, s), 1.95 (1H, br dd, J = 14.6, 6.2 Hz), (1H, m), 2.24 (1H, br dt, J = 15.2, 7.4 Hz), 3.20 (1H, br ddd, J = 10.0, 6.5, 2.3 Hz), 3.49 (2H, br dd, J = 7.8, 5.2 Hz), 3.70 (3H, s), 4.06 (1H, br dd, J = 12.6, 3.2 Hz), (4H, m), 5.32 (1H, br dd, J = 6.6, 5.2 Hz), 5.61 (1H, dt, J = 10.6, 6.5 Hz), 5.70 (1H, dt, J = 12.6, 6.5 Hz), 6.74 (2H, d, J = 8.6 Hz), (11H, m), (6H, m); S10 - -

12 13C NMR (100 MHz, CDCl3) δ 11.1, 11.9, 12.7, 18.0, 18.2, 18.3, 33.2, 40.3, 55.2, 59.6, 64.9, 70.8, 72.1, 78.9, 86.7, 113.7, 126.9, 127.7, 128.0, 128.4, 128.7, 129.2, 129.7, 130.9, 137.1, 144.3, 159.0; IR (neat) ν 2943, 2866, 1613, 1514, 1464, 1383, 1247, 1058, 882, 772, 705, 682, 594, 460 cm 1; HRMS (ESI) [M+Na] + calculated for [C57H84Na5Si2] + : , found: ; [α]d (c 1.03, H). (2E, 5S, 6S, 7R, 8Z)-6,8-Dimethyl-7,10-bis(triisopropylsiloxy)-1-(triphenylmethoxy)deca -2,8-dien-5-ol (12) TIPS PMB TIPS DDQ TIPS CH 2 Cl 2 /buffer = 5/1 Tr 0 C, 4 h H 96% TIPS Tr To a solution of 11 (450 mg, 0.50 mmol) in CH2Cl2 (4.1 ml) and ph 7.0 phosphate buffer solution (0.8 ml) was added 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ, 136 mg, 0.60 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 4 h at that temperature before being quenched with saturated aqueous Na2S23. After filtration with celite pad, the organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 50/1) to afford 12 (375 mg, 96%) as a pale yellow oil. 1H NMR (400 MHz, CDCl3) δ 0.76 (3H, d, J = 6.8 Hz) (42H, m), 1.86 (3H, s), 1.91 (1H, br dd, J = 13.6, 7.2 Hz), (1H, m), 2.30 (1H, br d, J = 14.4 Hz), 3.46 (1H, s), 3.55 (2H, d, J = 5.2 Hz) 3.61 (1H, br t, J = 6.8 Hz), 4.11 (1H, br dd, J = 12.4, 3.6 Hz), 4.28 (1H, dd, J = 12.4, 7.6 Hz), 4.69 (1H, d, J = 4.8 Hz), 5.45 (1H, br t, J = 6.4 Hz), 5.65 (1H, dt, J = 15.2, 5.2 Hz), (1H, m), (10H, m), 7.42 (5H, d, J = 7.6 Hz); 13C NMR (100 MHz, CDCl3) δ 12.0, 12.4, 13.0, 18.0, 18.1, 19.9, 38.0, 44.2, 59.6, 65.0, 73.1, 74.9, 86.7, 126.8, 127.7, 128.4, 128.7, 128.7, 129.5, 136.8, 144.3; IR (neat) ν 3480, 2943, 2866, 1490, 1464, 1383, 1104, 1055, 1013, 882, 763, 745, 705, S11 - -

13 682 cm 1; HRMS (ESI) [M+Na] + calculated for [C49H76Na4Si2] + : , found: ; [α]d (c 0.69, CHCl3). (2E, 5S, 6S, 7R, 8Z)-6,8-Dimethyl-7,10-bis(triisopropylsiloxy)-1-(triphenylmethoxy)deca-2, 8- dien-5-yl 1-azidocyclopropanecarboxylate (13) TIPS H (CCl) 2, DMF N 3 H benzene, 23 C TIPS 1 h TIPS Et TIPS 3 N, DMAP CH 2 Cl 2, 0 C, 20 min Tr 98% N Tr To a solution of 1-azidocyclopropanecarboxylic acid S3) (2.48 g, 19.5 mmol) and N,N-dimethylformamide (DMF, 0.15 ml, 1.95 mmol) in benzene (20 ml) was added oxalyl chloride (1.33 ml, 15.6 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 1 h at 23 C. anwhile, in the separated flask, 4-dimethylaminopyridine (DMAP, 236 mg, 1.95 mmol) was added to a solution of alcohol 12 (5.1 g, 6.5 mmol)and Et3N (5.44 ml, 39 mmol) in CH2Cl2 (20 ml). The carboxylic acid solution was transferred by cannula to the alcohol 12 solution at 0 C. The reaction mixture was stirred for 20 min at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 80/1) to afford 13 (5.67 g, 98%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ (45H, m), (4H, m), 1.84 (3H, s), (2H, m), (1H, m), 3.50 (2H, br t, J = 4.2 Hz), 4.13 (1H, dd, J = 12.0, 4.8 Hz), 4.35 (2H, dd, J = 14.4, 7.8 Hz), 4.80 (1H, br ddd, J = 10.2, 4.2, 2.4 Hz), 5.42 (1H, br t, J = 6.0 Hz), 5.57 (1H, br dt, J = 14.1, 6.0 Hz), 5.62 (1H, dt, J = 15.6, 4.8 Hz), 7.23 (3H, t, J = 7.8 Hz), 7.29 (6H, t, J = 7.2 Hz), 7.42 (6H, d, J = 7.2 Hz); S12 - -

14 13C NMR (100 MHz, CDCl3) δ 11.2, 12.0, 12.7, 16.5, 16.6, 18.0, 18.1, 18.2, 32.4, 41.2, 42.7, 59.4, 64.5, 72.1, 75.7, 86.8, 126.9, 127.4, 127.7, 128.2, 128.6, 130.1, 137.2, 144.2, 170.6; IR (neat) ν 2943, 2866, 2360, 2104, 1732, 1464, 1383, 1317, 1288, 1208, 1155, 1058, 882, 771, 745, 704, 681 cm 1; HRMS (ESI) [M+Na] + calculated for [C53H79N3Na5Si2] + : , found: ; [α]d (c 1.0, CHCl3). (2E, 5S, 6S, 7R, 8Z)-10-Hydroxy-6,8-dimethyl-7-(triisopropylsiloxy)-1-(triphenylmethoxy)d eca -2,8-dien-5-yl-1-azidocyclopropanecarboxylate (14) TIPS H TIPS HF Pyr TIPS CPh 3 THF, 23 C N 17 h 3 N 84% CPh 3 To a solution of azide 13 (5.47 g, 6.12 mmol) in THF (122 ml) was added 65% HF pyridine (6.9 ml, 245 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 17 h at 23 C before being quenched with saturated aqueous NaHC3 at 0 C. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 30/1) to afford 14 (3.77 g, 84%) as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 0.99 (21H, d, J = 3.6 Hz), 1.03 (3H, d, J = 6.4 Hz), (4H, m), 1.78 (3H, s), (2H, m), (1H, m), 3.46 (2H, s), (3H, m), 4.33 (1H, d, J = 8.4 Hz), (1H, m), 5.39 (1H, t, J = 6.8 Hz), 5.56 (2H, s), (10H, m), 7.35 (5H, d, J = 7.6 Hz); 13C NMR (100 MHz, CDCl3) δ 11.4, 12.6, 16.6, 16.7, 18.1, 32.9, 41.6, 42.7, 58.6, 64.4, 72.4, 76.0, 86.8, 126.9, 127.0, 127.7, 128.5, 130.3, 139.8, 144.1, 171.0; IR (neat) ν 3446, 2944, 2866, 2106, 1729, 1448, 1384, 1318, 1287, 1209, 1158, 1058, 883, 746, 706, 681, 633, 420 cm 1; HRMS (ESI) [M+Na] + calculated for [C44H59N3Na5Si] + : , found: ; [α]d (c 1.08, CHCl3). S13 - -

15 (2E, 5S, 6S, 7R, 8Z)- 11-(2-Hydroxy-5-methoxy-3-nitrophenyl)-6,8-dimethyl- 11- (phenylsulfonyl)-7-(triisopropylsiloxy)-1-(triphenylmethoxy)undeca-2,8-die n-5-yl-1-azidocyclopropanecarboxylate (17) H TIPS N 3 14 I 1) I 2, Ph 3 P imidazole TIPS benzene CPh 3 23 C, 30 min N 3 15 CPh 3 2) LHMDS, Ph 2 S N 2 N 2 16 H S 2 Ph H TIPS THF,!90 C to!65 C 18 h CPh 78% (2 steps) 3 N 3 17 To a solution of alcohol 14 (328 mg, 0.44 mmol), imidazole (181 mg, 2.66 mmol) and iodine (563 mg, 2.22 mmol) in benzene (7.0 ml) was added dropwise to a solution of Ph3P (466 mg, 1.78 mmol) in benzene (2.0 ml) over 30 min at 23 C under argon atmosphere. The resulting mixture was quenched with ph 7.0 phosphate buffer solution and saturated aqueous Na2S23. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. (Caution : Complete dryness induce critical decomposition.) The residue with small amount of ethyl acetate was directly purified by flash column chromatography (Si2, hexane/ethyl acetate = 20/1) to afford unstable iodide 15 (378 mg, quantitative yield) as a pale yellow oil. To a solution of aromatic segment 16 (718 mg, 2.22 mmol) in THF (34 ml) was added Lithium bis(trimethylsilyl)amide (LHMDS, 4.7 ml, 1.0 M THF solution, 4.66 mmol) at 90 C under argon atmosphere. The reaction mixture was stirred for 40 min at that temperature. The resulting mixture was transferred by cannula to the solution of unstable iodide 15 (378 mg, 0.44 mmol) in THF (10 ml) at 90 C followed by raising the temperature until 65 C over 2 h. The reaction mixture was stirred for additional S14 - -

16 18 h at 65 C before being quenched with cooled ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 20/1) to afford 17 (361 mg, 78%, 2 steps) as a yellow amorphous powder. 1H NMR (400 MHz, CDCl3, mixture of diastereomer) δ (24H, m), (4H, m), 1.62 (3H, s), (2H, m), (1H, m), 2.55 (1H, br t, J = 13.2 Hz), 3.24 (1H, br t, J = 11.2 Hz), (2H, m), 3.59 (3H, s), 4.32 (1H, br d, J = 7.6 Hz), (3H, m), (2H, m), (22H, m), (1H, br s); 13C NMR (100 MHz, CDCl3, mixture of diastereomer) δ 11.2, 11.3, 12.7, 16.6, 16.4, 18.1, 18.1, 18.2, 18.2, 25.9, 26.6, 32.6, 32.9, 41.5, 42.7, 55.9, 56.0, 64.5, 64.8, 72.1, 75.1, 76.0, 86.8, 86.9, 106.9, 107.3, 124.2, 124.6, 126.4, 126.9, 127.0, 127.4, 127.8, 128.6, 128.7, 128.9, 130.5, 130.7, 133.1, 133.8, 133.9, 137.5, 137.7, 141.1, 144.2, 144.2, 148.5, 148.6, 152.1, 170.7; IR (neat) ν 3022, 2944, 2866, 2106, 1729, 1596, 1542, 1466, 1448, 1430, 1383, 1320, 1254, 1215, 1150, 1085, 1056, 883, 760, 706, 687, 602, 555, 518, 443 cm 1; HRMS (ESI) [M+Na] + calculated for [C58H70N4Na10SSi] + : , found: ; [α]d (c 2.93, CHCl3). (2E, 5S, 6S, 7R, 8Z)- 11-[2-(tert-Butoxycarbonyloxy)-5-methoxy-3-nitrophenyl]-6,8- dimethyl- 11-(phenylsulfonyl)-7-(triisopropylsiloxy)-1-(triphenylmethoxy)u ndeca-2,8-dien-5-yl 1-azidocyclopropanecarboxylate (18) TIPS N 3 S 2 Ph H N 2 (Boc) 2 DMAP TIPS CH 2 Cl 2, 23 C Tr 30 min 17 96% N 3 18 N 2 S 2 Ph Boc Tr To a solution of azide 17 (956 mg, 0.92 mmol) and 4-dimethylaminopyridine (DMAP, 224 mg, 1.83 mmol) in CH2Cl2 (63 ml) was added (Boc)2 (0.66 ml, 2.75 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 30 min at 23 C before S15 - -

17 being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 5/1) to afford 18 (1.01 g, 96%) as a yellow amorphous powder. 1H NMR (400 MHz, CDCl3, mixture of diastereomer) δ (24H, m), (4H, m), 1.49 (6H, s), 1.53 (3H, s), 1.69 (3H, s), (2H, m), (1H, m), 2.62 (1H, br t, J = 14.0 Hz), (1H, m), 3.57 (2H, t, J = 4.8 Hz), 3.65 (3H, s), 4.40 (1H, br d, J = 7.6 Hz), 4.59 (1H, br d, J = 12.0 Hz), (2H, m), (2H, m), (9H, m), (8H, m), (3H, m), 7.64 (2H, d, J = 7.6 Hz); 13C NMR (100 MHz, CDCl3, mixture of diastereomer) δ 11.0, 11.3, 12.7, 14.1, 14.2, 16.6, 16.6, 18.1, 18.1, 18.2, 18.2, 20.7, 21.0, 22.6, 25.2, 26.9, 27.5, 31.5, 32.9, 33.0, 34.5, 34.6, 41.7, 41.8, 42.7, 42.8, 56.0, 56.1, 60.3, 61.7, 62.2, 64.5, 64.7, 72.2, 75.5, 75.7, 85.2, 85.6, 86.8, 86.9, 110.6, 111.2, 119.6, 120.1, 120.5, 126.9, 126.9, 127.8, 128.6, 128.7, 128.9, 129.1, 129.6, 130.5, 130.8, 131.9, 134.0, 137.2, 137.3, 141.2, 142.3, 144.2, 149.6, 156.6, 170.6, 170.6; IR (neat) ν 2944, 2867, 1771, 1727, 1621, 1584, 1543, 1448, 1315, 1211, 1146, 1058, 1003, 910, 882, 820 cm 1; HRMS (ESI) [M+Na] + calculated for [C63H78N4Na12SSi] + : , found: ; [α]d (c 0.84, CHCl3). (2E, 5S, 6S, 7R, 8Z)- 11-[2-(tert-Butoxycarbonyloxy)-5-methoxy-3-nitrophenyl]-6,8- dimethyl- 11-(phenylsulfonyl)-7-(triisopropylsiloxy)-1-(triphenylmethoxy)u ndeca-2,8-dien-5-yl 1-aminocyclopropanecarboxylate (19) TIPS N 3 N 2 S 2 Ph Boc HS Et 3 N SH TIPS N 2 S 2 Ph Boc H, 23 C 18 h CPh 3 87% CPh 3 H 2 N To a solution of azide 18 (1.84 g, 1.61 mmol) and Et3N (11.2 ml, 80.5 mmol) in H S16 - -

18 (37 ml) was added 1,3-propanedithiol (8.1 ml, 80.5 mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 15 h at that temperature. After volatile materials were removed in vacuo, ph 7.0 phosphate buffer solution was added to the crude materials. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 2/1) to afford 19 (1.56 g, 87%) as a yellow amorphous powder. 1H NMR (400 MHz, CDCl3, mixture of diastereomer) δ 0.97 (3H, d, J = 6.4 Hz), 1.06 (21H, d, J = 5.6 Hz), (3H, m), 1.45 (1H, s), 1.49 (6H, s), 1.70 (3H, s), (6H, m), (1H, m), 2.61 (1H, br t, J = 13.6 Hz), (1H, m), 3.57 (2H, br t, J = 6.4 Hz), 3.65 (3H, s), (1H, m), (1H, m), (1H, m), 4.78 (1H, br d, J = 8.8 Hz), (1H, m), (2H, m), (9H, m), (8H, m), (3H, m), 7.63 (2H, d, J = 7.6 Hz); 13C NMR (100 MHz, CDCl3, mixture of diastereomer) δ 11.2, 12.7, 18.0, 18.2, 18.2, 20.7, 25.2, 27.5, 32.8, 34.6, 35.8, 41.6, 55.9, 61.6, 64.5, 74.4, 85.2, 86.8, 111.0, 120.2, 126.9, 127.3, 127.8, 128.6, 128.6, 128.7, 128.9, 129.0, 130.0, 134.0, 137.2, 142.2, 144.1, 144.2, 149.6, 156.5, 175.8; IR (neat) ν 3884, 3022, 2944, 2867, 1771, 1717, 1585, 1543, 1448, 1312, 1287, 1214, 1146, 1056, 1003, 882, 753, 705, 599, 525 cm 1; HRMS (ESI) [M+Na] + calculated for [C63H80N2Na12SSi] + : , found: ; [α]d (c 1.18, CHCl3). (2E, 5S, 6S, 7R, 8Z)- 11-[2-(tert-Butoxycarbonyloxy)-5-methoxy-3-nitrophenyl]-6,8- dimethyl- 11-(phenylsulfonyl)-7-(triisopropylsiloxy)-1-(triphenylmethoxy)u ndeca-2,8-dien-5-yl 1-(cyclohex-1-enecarboxamido)cyclopropanecarboxylate (20) N 2 S 2 Ph Boc TIPS CPh 3 H 2 N 19 H EDCI HCl, DMAP CH 2 Cl 2, 23 C 21 h 78% TIPS H N N 2 S 2 Ph Boc CPh 3 20 S17 - -

19 To a solution of amine 19 (918 mg, 0.82 mmol) and 4-dimethylaminopyridine (DMAP, 802 mg, 6.57 mmol) in CH2Cl2 (17 ml) were added a solution of 1-cyclohexenecarboxylic acid (311 mg, 2.46 mmol) in CH2Cl2 (10 ml) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI HCl, 630 mg, 3.28 mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 21 h at that temperature before being quenched with saturated aqueous NaHC3. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaHC3 (3 times) and saturated aqueous NaCl (2 times). The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 3/1) to afford 20 (780 mg, 78%) as a yellow amorphous powder. 1H NMR (400 MHz, CDCl3, mixture of diastereomer) δ (24H, m), (3H, m), (16H, m), (1H, m), (4H, m), (6H, m), 2.62 (1H, br t, J = 13.6 Hz), 3.41 (1H, br t, J = 12.0 Hz), (1H, m), 3.70 (3H, s), 4.32 (1H, br d, J = 10.4 Hz), 4.51 (1H, br d, J = 5.2 Hz), (2H, m), (2H, m), 6.52 (2H, br s), 6.95 (1H, br s), (7H, m), (12H, m); 13C NMR (100 MHz, CDCl3, mixture of diastereomer) δ 10.9, 12.8, 14.1, 17.4, 18.1, 18.2, 18.2, 20.7, 21.1, 21.5, 21.8, 22.0, 22.6, 24.0, 24.1, 25.3, 26.1, 31.6, 33.2, 33.8, 34.5, 34.7, 56.0, 62.3, 65.0, 75.1, 86.8, 111.0, 120.4, 126.9, 127.6, 127.8, 128.7, 128.9, 130.0, 133.2, 133.6, 134.1, 137.3, 142.3, 143.9, 144.3, 156.6, 169.3, 172.0; IR (neat) ν 3398, 2941, 2866, 1734, 1669, 1639, 1541, 1491, 1446, 1311, 1190, 1056, 1009, 924, 883, 754, 705, 601 cm 1; MS (ESI); (C65H80N2Na11SSi: Boc group was deprotected in Mass) [α]d (c 1.45, CHCl3). (2E, 5S, 6S, 7R, 8Z)- 11-(2-Hydroxy-5-methoxy-3-nitrophenyl)-6,8-dimethyl-7- (triisopropylsiloxy)-1-(triphenylmethoxy)undeca-2,8-dien-5-yl-1-(cyclohex- 1-enecarboxamido)cyclopropanecarboxylate (22) S18 - -

20 TIPS H N N 2 S 2 Ph Boc CPh ) pyrrolidine CH 2 Cl 2, 23 C 5 h TIPS H N S 2 Ph H CPh 3 21 N 2 2) NaBH 4 EtH, 50 C, 21 h 57% (2 steps) TIPS H N H CPh 3 22 To a solution of amide 20 (780 mg, 0.64 mmol) in CH2Cl2 (32 ml) was added pyrrolidine (9.4 ml, mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 5 h at that temperature before being quenched with saturated aqueous NH4Cl. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NH4Cl (3 times). The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 2/1) N 2 to afford 21 (698 mg, 97%) as a yellow amorphous powder. To a solution of 21 (143 mg, 0.13 mmol) in EtH (9.4 ml) was added NaBH4 (192 mg, 5.06 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 21 h at 50 C before being quenched with saturated aqueous NH4Cl. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 10/1) to afford 22 (125 mg, 57%, 2 steps) as a yellow amorphous powder. 1H NMR (400 MHz, CDCl3) δ (24H, m), (2H, m), (7H, m), 1.81 (3H, s), (5H, m), (2H, m), (1H, m), (2H, m), 3.52 (2H, d, J = 4.0 Hz), 3.75 (3H, s), 4.36 (1H, d, J = 9.2 Hz), (1H, m), 5.25 (1H, br t, J = 6.4 Hz), (2H, m), 6.10 (1H, s), 6.52 (1H, br s), 7.05 (1H, d, J = 3.2 Hz), (10H, m), 7.35 (1H, d, J = 3.2 Hz), (6H, m); 13C NMR (100 MHz, CDCl3) δ 11.1, 12.6, 14.2, 17.2, 18.1, 18.2, 21.5, 22.0, 24.1, 25.3, 27.7, 30.6, 32.4, 33.9, 40.9, 55.8, 64.7, 72.1, 74.7, 86.8, 103.4, 126.1, 126.9, 127.4, 127.7, 128.3, 128.6, 129.6, 133.0, 133.2, 133.7, 134.0, 137.7, 144.2, 148.8, 151.8, 169.2, 171.7; S19 - -

21 IR (neat) ν 3422, 3338, 2941, 2865, 1727, 1668, 1634, 1539, 1493, 1466, 1430, 1374, 1327, 1255, 1174, 1055, 999, 883, 758, 705 cm 1; HRMS (ESI) [M+Na] + calculated for [C59H76N2Na9Si] + : , found: ; [α]d (c 1.33, CHCl3). (2E, 5S, 6S, 7R, 8Z)- 11-[2-(Allyloxycarbonyloxy)-5-methoxy-3-nitrophenyl]-1-hydroxy -6,8-dimethyl-7-(triisopropylsiloxy)undeca-2,8-dien-5-yl-1-(cyclohex-1-ene carboxamido)cyclopropanecarboxylate (24) TIPS H N H CPh 3 22 N 2 1) AllocCl, Et 3 N CH 2 Cl 2, 23 C 40 min TIPS H N N 2 Alloc CPh ) TsH H 2 H, 23 C 1 h 94% (2 steps) TIPS H N 24 N 2 Alloc H To a solution of 22 (36 mg, mmol) and Et3N (127 µl, 0.91 mmol) in CH2Cl2 (1.8 ml) was added chloroformic acid allyl ester (AllocCl, 50.3 µl, 0.48 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 40 min at 23 C before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration to afford the crude material 23 (41.3 mg) as a pale yellow amorphous powder. To a solution of crude material 23 (41.3 mg) in H (3.7 ml) was added p-toluenesulfonic acid monohydrate (TsH H2, 5.6 mg, mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 1 h at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under S20 - -

22 reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 1/4) to afford 24 (28.3 mg, 94%, 2 steps) as a pale yellow amorphous powder. 1H NMR (400 MHz, CDCl3) δ (25H, m), (1H, m), (5H, m), 1.76 (4H, s), 1.99 (2H, br d, J = 11.6 Hz), (6H, m), (1H, m), (2H, m), 3.61 (1H, br s), 3.85 (3H, s), 4.01 (2H, br d, J = 5.2 Hz), 4.33 (1H, d, J = 8.4 Hz), 4.76 (2H, d, J = 7.6 Hz), 4.84 (1H, br d, J = 11.2 Hz), 5.21 (1H, br t, J = 6.4 Hz), 5.34 (1H, d, J = 10.4 Hz), (2H, m), 5.61 (1H, dt, J = 14.8, 5.6 Hz), (1H, m), 6.23 (1H, s), 6.62 (1H, br s), 7.00 (1H, br d, J = 2.8 Hz), 7.46 (1H, br d, J = 3.2 Hz); 13C NMR (100 MHz, CDCl3) δ 11.3, 12.7, 17.0, 17.5, 18.1, 18.2, 21.4, 22.0, 24.1, 25.4, 28.2, 30.3, 32.6, 33.7, 41.6, 56.0, 63.3, 70.0, 72.2, 74.3, 108.1, 119.7, 121.8, 125.6, 126.8, 130.8, 132.9, 133.6, 134.6, 136.2, 138.1, 138.3, 142.1, 152.6, 157.0, 169.7, 171.5; IR (neat) ν 3332, 2942, 2866, 1771, 1726, 1664, 1630, 1540, 1462, 1349, 1288, 1231, 1203, 1083, 1058, 995, 941, 883, 757, 681, 600, 445 cm 1; HRMS (ESI) [M+Na] + calculated for [C44H66N2Na11Si] + : , found: ; [α]d (c 0.31, CHCl3). (3E, 6S, 7S, 8R, 9Z)-12-[2-(Allyloxycarbonyloxy)-5-methoxy-3-nitrophenyl] -7,9-dimethyl-8-(triisopropylsiloxy)dodeca-1,3,9-trien-6-yl-1-(cyclohexenyl methylamino)cyclopropanecarboxylate (26) TIPS H N 24 N 2 Alloc 1) Mn 2 H CH 2 Cl 2, 23 C 40 min TIPS H N 25 N 2 Alloc H 2) [Ph 3 P-CH 3 ]I t BuK THF, 0 C, 30 min 74% (2 steps) TIPS H N 26 N 2 Alloc To a solution of alcohol 24 (21 mg, mmol) in CH2Cl2 (5 ml) was added excess amount of Mn2 at 23 C under argon atmosphere. The reaction mixture was stirred for 40 min at that temperature before being removed Mn2 by filtration with celite pad. S21 - -

23 The organic materials were concentrated under reduced pressure to afford crude material of 25 (19 mg) as a pale yellow amorphous powder. To a solution of methyltriphenylphosphonium iodide (46.2 mg, mmol) in THF (1.5 ml) was added potassium tert-butoxide (10 mg, mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 30 min at that temperature. The resulting mixture was transferred by cannula to the solution of aldehyde 25 (19 mg, mmol) in THF (0.5 ml) at that temperature. The reaction mixture was stirred for additional 30 min at that temperature before being quenched with saturated aqueous NH4Cl at 0 C. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by preparative TLC (Si2, hexane/ethyl acetate = 3/1) to afford 26 (15.4 mg, 74%, 2 steps) as a pale yellow amorphous powder. 1H NMR (400 MHz, CDCl3) δ (25H, m), (6H, m), 1.79 (3H, br s), (10H, m), (2H, m), 3.85 (3H, s), 4.33 (1H, d, J = 8.8 Hz), 4.72 (1H, br dt, J = 9.6, 2.4 Hz), 4.76 (2H, d, J = 6.0 Hz), 4.97 (1H, d, J = 10.0 Hz), 5.07 (1H, d, J = 16.8 Hz), 5.19 (1H, br t, J = 6.8 Hz), 5.35 (1H, d, J = 10.4 Hz), 5.44 (1H, br dd, J = 16.0, 1.2 Hz), 5.51 (1H, dd, J = 14.4, 6.8 Hz), (2H, m), 6.12 (1H, s), (1H, m), 6.58 (1H, br s), 7.01 (1H, d, J = 2.8 Hz), 7.46 (1H, d, J = 3.2 Hz); 13C NMR (100 MHz, CDCl3) δ 11.0, 12.7, 17.2, 17.3, 18.1, 18.2, 21.5, 22.1, 24.2, 25.4, 28.1, 30.3, 32.5, 33.9, 41.1, 56.0, 69.9, 72.2, 74.9, 108.1, 115.9, 119.7, 121.8, 125.4, 129.9, 130.8, 133.3, 133.6, 133.7, 136.2, 136.7, 138.3, 138.4, 142.1, 152.6, 157.0, 169.2, 171.8; IR (neat) ν 3327, 3089, 2941, 2867, 1771, 1726, 1665, 1631, 1540, 1459, 1349, 1203, 1060, 1004, 945, 883, 758, 682 cm 1; HRMS (ESI) [M+Na] + calculated for [C44H66N2Na10Si] + : , found: ; [α]d (c 0.87, CHCl3). (3E, 6S, 7R, 8R, 9Z)-12-[2-(Allyloxycarbonyloxy)-5-methoxy-3-nitrophenyl]-8-hydroxy -7,9-dimethyl-dodeca-1,3,9-trien-6-yl-1-(cyclohex-1-enecarboxamido)cyclo propanecarboxylate (27) S22 - -

24 TIPS H N 26 N 2 Alloc 46% HF CN 23 C, 16 h 68% (borsm : 91%) H N H 27 N 2 Alloc To a solution of diene 26 (80 mg, mmol) in CN (8.0 ml) was added 46% HF (0.8 ml, 44 mmol) at 0 C under argon atmosphere. The reaction mixture was stirred for 16 h at 23 C before being quenched with saturated aqueous NaHC3 at 0 C. The organic materials were extracted with ethyl acetate (3 times) and chloroform (once). The combined organic phase was dried over anhydrous MgS4 and concentrated under reduced pressure after filtration. The residue was purified by flash column chromatography (Si2, hexane/ethyl acetate = 1/1) to provide alcohol 27 (43.8 mg, 68%) as a pale yellow amorphous powder and recovered 26 (21 mg, 26%). 1H NMR (400 MHz, CDCl3) δ 0.92 (3H, d, J = 7.2 Hz), (2H, m), (9H, m), (1H, m), (4H, m), (5H, m), (2H, m), 3.85 (3H, s), 4.33 (1H, br d, J = 4.8 Hz), 4.77 (2H, dt, J = 6.0, 1.2 Hz), 4.82 (1H, dt, J = 7.6, 4.0 Hz), 4.98 (1H, d, J = 10.4 Hz), 5.08 (1H, d, J = 16.8 Hz), 5.20 (1H, br t, J = 14.4 Hz), 5.35 (1H, dd, J = 10.4, 0.8 Hz), 5.44 (1H, dt, J = 16.8, 1.2 Hz), (1H, m), (2H, m), (2H, m), (1H, m), 7.09 (1H, d, J = 3.2 Hz), 7.45 (1H, d, J = 3.2 Hz); 13C NMR (100 MHz, CDCl3) δ 10.2, 17.4, 17.5, 20.0, 21.5, 22.0, 24.1, 25.4, 27.7, 30.4, 33.8, 34.4, 40.9, 56.1, 70.0, 75.8, 108.0, 115.9, 119.8, 122.0, 125.1, 129.4, 130.8, 133.0, 133.8, 134.3, 136.2, 136.7, 138.1, 138.7, 142.0, 152.6, 156.9, 169.5, 172.5; IR (neat) ν 3416, 3089, 2935, 1770, 1725, 1664, 1629, 1538, 1448, 1352, 1283, 1203, 1038, 1005, 946, 861, 823, 769, 738, 704 cm 1; HRMS (ESI) [M+Na] + calculated for [C36H46N2Na10] + : , found: ; [α]d (c 1.0, H). (3E, 6S, 7S, 8R, 9Z)-12-[2-(Allyloxycarbonyloxy)-5-methoxy-3-nitrophenyl]- 7,9-dimethyl-8-(triethylsiloxy)dodeca-1,3,9-trien-6-yl-1-(cyclohexenylmeth ylamino)cyclopropanecarboxylate (28) S23 - -

25 H N H 27 N 2 Alloc TESTf i Pr 2 EtN CH 2 Cl 2, 23 C 30 min 99% TES H N 28 N 2 Alloc To a solution of alcohol 27 (17 mg, mmol) in CH2Cl2 (1.5 ml) were added i Pr2EtN (44.5 µl, mmol) and triethylsilyl triflate (TESTf, 29 µl, mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 30 min at that temperature before being quenched with ph 7.0 phosphate buffer solution. The organic materials were extracted with ethyl acetate (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by preparative TLC (Si2, hexane/ethyl acetate = 4/1) to afford 28 (19.7 mg, 99%) as a pale yellow amorphous powder. 1H NMR (400 MHz, CDCl3) δ 0.52 (6H, q, J = 7.6 Hz), 0.91 (9H, t, J = 7.6 Hz), 1.13 (2H, d, J = 2.0 Hz), (2H, m), (6H, m), 1.75 (3H, s), (1H, m), (8H, m), (1H, m), (2H, m), 3.85 (3H, s), 4.19 (1H, d, J = 8.0 Hz), 4.70 (1H, ddd, J = 9.2, 4.0, 2.8 Hz), 4.76 (2H, d, J = 6.0 Hz), 4.96 (1H, d, J = 10.4 Hz), 5.07 (1H, d, J = 17.2 Hz), 5.18 (1H, t, J = 6.4 Hz), 5.34 (1H, dd, J = 10.4, 0.8 Hz), 5.44 (1H, dd, J = 17.2, 1.2 Hz), (1H, m), (2H, m), 6.14 (1H, s), 6.25 (1H, dt, J = 17.2, 10.4 Hz), (1H, m), 7.05 (1H, d, J = 2.8 Hz), 7.46 (1H, d, J = 2.8 Hz); 13C NMR (100 MHz, CDCl3) δ 4.8, 5.8, 6.8, 10.7, 17.2, 18.6, 21.5, 22.1, 24.2, 25.3, 28.0, 30.3, 32.7, 33.9, 40.8, 56.0, 69.9, 71.7, 75.1, 108.1, 115.8, 119.7, 121.8, 125.1, 129.9, 130.8, 133.3, 133.5, 133.7, 136.2, 136.8, 138.4, 138.5, 142.1, 152.6, 156.9, 169.2, 171.8; IR (neat) ν 3671, 3327, 3088, 2937, 1771, 1732, 1667, 1633, 1584, 1537, 1504, 1494, 1455, 1361, 1294, 1203, 1066, 1006, 946, 853, 823, 742, 611 cm 1; HRMS (ESI) [M+Na] + calculated for [C42H60N2Na10Si] + : , found: ; [α]d (c 1.92, H). (3E, 6S, 7S, 8R, 9Z)-12-{2-Hydroxy-5-methoxy-3-[(3R, 4E)-3-methoxy- hepta-4,6-dienoylamino]-phenyl}-7,9-dimethyl-8-(triethylsiloxy)dodeca-1,3,9-trien-6-yl-1-(cyclohexenylmethylamino)cyclopropanecarboxylate (30) S24 - -

26 TES H N 28 N 2 Alloc 1) NaBH 4, S 8 THF, 50 C, 2.5 h TES H N 29 H NH 2 H 2) NH 42 TES i Pr 2 EtN, BPCl 3) K 2 C 3 H toluene, 23 C, 8 h N H, 23 C 10 min 79% (3 steps) S4 TES NH H H N 30 To a solution of diene 28 (21.7 mg, mmol) in THF (3.8 ml) were added sulfer (31.2 mg, mmol) and NaBH4 (21 mg, mmol) at 23 C under argon atmosphere. The reaction mixture was stirred for 2.5 h at 50 C before being quenched with saturated aqueous NaHC3. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous MgS4 and concentrated under reduced pressure after filtration to afford unstable crude material of 29 as a pale yellow amorphous powder. To a solution of crude material of 29 in toluene (3.8 ml) was added i Pr2EtN (248 µl, 1.39 mmol) at 23 C under argon atmosphere. The resulting mixture was transferred by cannula to the stirred solution of C1 C6 segment 42 (26 mg, mmol) and bis(2-oxo-3-oxazolidinyl) phosphinic chloride (BPCl, 21.2 mg, mmol) in THF (10 ml) at 23 C. The resulting mixture was stirred for 8 h at that temperature before being quenched with saturated aqueous NaHC3. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration to afford crude materials of S4 as a pale yellow amorphous powder. To a solution of crude materials of S4 in H (4.0 ml) was added K2C3 (40 mg) at S25 - -

27 23 C under argon atmosphere. The reaction mixture was stirred for 10 min at that temperature before being quenched with saturated aqueous NH4Cl. The organic materials were extracted with chloroform (3 times) and washed with saturated aqueous NaCl. The combined organic phase was dried over anhydrous Na2S4 and concentrated under reduced pressure after filtration. The residue was purified by preparative TLC (Si2, hexane/ethyl acetate = 2/1) to afford 30 (17.6 mg, 79%, 3 steps) as a pale yellow amorphous powder. 1H NMR (400 MHz, CDCl3) δ 0.53 (6H, t, J = 8.0 Hz), (9H, m), 1.13 (2H, d, J = 3.2 Hz), (9H, m), 1.75 (3H, s), 1.98 (1H, q, J = 6.8 Hz), (8H, m), (1H, m), (3H, m), 3.37 (3H, s), 3.73 (3H, s), (1H, m), 4.30 (1H, d, J = 7.2 Hz), 4.74 (1H, q, J = 6.0 Hz), 4.95 (1H, d, J = 10.0 Hz), 5.06 (1H, d, J = 16.4 Hz), (3H, m), (2H, m), 6.00 (1H, dd, J = 15.2, 10.4 Hz), (4H, m), 6.57 (3H, s), 8.16 (1H, br s), 8.66 (1H, s); 13C NMR (100 MHz, CDCl3) δ 4.8, 6.9, 10.8, 17.2, 17.3, 18.8, 21.5, 22.1, 24.2, 25.3, 28.2, 29.7, 31.6, 33.0, 33.9, 41.2, 42.9, 55.7, 56.5, 71.3, 75.5, 78.6, 105.5, 113.2, 115.4, 119.2, 126.4, 126.6, 130.2, 130.8, 133.1, 133.3, 133.5, 133.7, 134.6, 135.6, 137.1, 137.5, 140.6, 152.9, 169.5, 170.3; IR (neat) ν 3328, 2932, 2876, 2360, 1726, 1660, 1627, 1488, 1347, 1297, 1236, 1176, 1065, 1006, 841, 742 cm 1; HRMS (ESI) [M+Na] + calculated for [C46H68N2Na8Si] + : , found: ; [α]d (c 0.47, H). (3E, 6S, 7S, 8R, 9Z)-12-{2,5-Dihydroxy-3-[(3R, 4E)-3-methoxy-hepta- 4,6-dienoylamino]-phenyl}-7,9-dimethyl-8-(triethylsiloxy)dodeca-1,3,9-trie n-6-yl-1- (cyclohexenylmethylamino)cyclopropanecarboxylate (31) S26 - -