SUEMENTARY INRMATIN - Nature Research · mixture was stirred at 5 °C for 5 h and then cooled to 78 °C. MeI (13.4 ml, 30.6 g, 215 mmol, 3.5 eq.) was subsequently added very slowly

NATURE CHEMISTRY | www.nature.com/naturechemistry 1

SUPPLEMENTARY INFORMATIONDOI: 10.1038/NCHEM.2336

An Eight-Step Synthesis of Epicolactone

Reveals its Biosynthetic Origin

Pascal Ellerbrock,* Nicolas Armanino,* Marina K. Ilg, Robert Webster and Dirk Trauner

Department of Chemistry and Center for Integrated Protein Science, Ludwig Maximilians

University Munich, Butenandtstraße 5-13, 81377 Munich, Germany

* These authors contributed equally to this work.

Supplementary Information

Table of Content

1. General Procedures .......................................................................................................... S1

2. Chromatography ............................................................................................................... S1

3. Nuclear Magnetic Resonance Spectroscopy (NMR) ...................................................... S2

4. Mass Spectrometry ........................................................................................................... S2

5. Solvents and Reagents ...................................................................................................... S2

6. Melting Points ................................................................................................................... S3

7. Infrared Spectroscopy (IR) .............................................................................................. S3

8. LC/MS ................................................................................................................................ S3

9. Additional Information .................................................................................................... S4

10. Experimental procedures ................................................................................................. S5

11. X-Ray Single Crystal Structure ..................................................................................... S25

12. NMR spectra ................................................................................................................... S34

An eight-step synthesis of epicolactone reveals its biosynthetic origin

© 2015 Macmillan Publishers Limited. All rights reserved



SUPPORTING INFORMATION S1

1. General Procedures

All air and/or moisture sensitive reactions were conducted under inert gas atmosphere

(nitrogen). Glassware was stored in an oven (130 °C) and heat-gun dried (650 °C) under

high-vacuum prior to use (three times, in between filled with nitrogen). Liquids and solutions

were transferred via three times nitrogen-flushed syringes with oven-dried stainless steel

cannulas (130 °C). Solids were added under counter flow of nitrogen (Schlenk technique).

Solutions were concentrated under reduced pressure by rotary evaporation at 30 °C with

Heidolph Laborota 4000 efficient employing a vacuubrand PC 3001 pump.

membrane pump for rotary evaporation (VACUUBRAND): 4 mbar

oil pump for high-vacuum: 0.006 mbar

Yields, unless noted otherwise, are isolated yields. All reactions were magnetically stirred and

monitored either by LC/MS or TLC (vide infra).

2. Chromatography

Thin-Layer Chromatography (TLC): TLC plates (silica gel 60) on glass with fluorescence

indicator F-254 (MERCK) were used. Eluents were of HPLC grade. TLCs were analyzed by

UV light ( = 254 nm), if applicable, or immersing in staining solutions. After staining, TLC

plates were heated with a heat-gun at 300 °C until dry.

Staining solutions:

KMnO4-solution: 3 g KMnO4, 20 g K2CO3, 5 ml 5% NaOH (aq), 300 ml dist. H2O

CAN-solution: 6.25 g molybdophosphoric acid, 2.50 g Ce(SO4)2·4H2O and 15 ml

sulfuric acid (97%) in 230 ml dist. H2O

Flash Column Chromatography: Purification by flash column chromatography was achieved

using silica gel 60 (40-63 m, MERCK) with HPLC grade solvents. For purification with C18

silica gel flash column chromatography, silica gel (preparative C18, 125 Å, 55105 m,

Waters Corp.) was suspended in HPLC grade MeCN, the column packed and the silica gel

washed twice with the starting eluent. The volume of silica gel used is provided in (height x

diameter column). The compounds were loaded in dioxane or THF and fractions analyzed by

LC/MS (vide infra). The combined fractions were extracted with EtOAc three times, the

combined organic phases dried over Na2SO4 and concentrated under reduced pressure.





3. Nuclear Magnetic Resonance Spectroscopy (NMR)

All nuclear magnetic resonance spectra were recorded either on Varian VNMRS 300,

VNMRS 400, INOVA 400, VNMRS 600 or a Bruker AVANCEIIIHD 400 spectrometer.

NMR spectra were referenced to the undeuterated solvent signal (CDCl3: 1H NMR 7.26

ppm, 13C NMR 77.16 (t) ppm; (D3C)2CO: 1H NMR 2.05 (quint) ppm, 13C NMR 29.84

(sept) ppm; [D6]DMSO: 1H NMR 2.50 (quint) ppm, 13C NMR 39.52 (sept) ppm;

[D7]DMF: 1H NMR 2.75 (quint) ppm, 13C NMR 29.76 (sept) ppm). Spectral data is

provided in ppm from downfield to upfield. The following abbreviations are utilized in the

analysis of NMR spectra: s = singlet, d = doublet, t = triplet, q = quartet, quint = quintet,

sept = septet, m = multiplet, brs = broad singlet. Combination of these abbreviations is applied

whenever more than one coupling is observed. Data is provided in the following order:

chemical shift in ppm (multiplicity, coupling constant J in Hz, signal integration). Unless

noted differently, coupling constants are 3JH,H in 1H NMR. NMR spectra were recorded at

T = 27 °C. Analysis of all spectra was performed with MestReNova version 8.1 by Mestrec

Laboratories.

4. Mass Spectrometry

High resolution (HRMS) and low resolution (LRMS) mass spectra were recorded on a

Finnigan MAT 90 and a Finnigan MAT 95 instrument. Ionization of the samples was

achieved using electrospray ionization (ESI) or electron ionization (EI).

5. Solvents and Reagents

Commercial reagents and dry solvents over molecular sieves were used as purchased from

Aldrich or Acros Organics with the following exceptions. Tetrahydrofuran (THF) and

diethylether (Et2O) were pre-dried over CaCl2 and distilled over sodium and benzophenone

under a nitrogen atmosphere before use. Triethylamine (NEt3), diisopropylamine (DIPA) and

diisopropylethylamine (DIPEA) were distilled over CaH2 under a nitrogen atmosphere prior

to use. Solvents were stored under nitrogen.





6. Melting Points

Melting points were measured on a BÜCHI B-540 melting point apparatus and are

uncorrected.

7. Infrared Spectroscopy (IR)

Infrared (IR) spectra were recorded on a Perkin Elmer Spectrum BX-59343 instrument with a

Smiths Detection DuraSamplIR II Diamond ATR sensor for detection in the range from

ῦ = 4500 cm–1 to ῦ = 600 cm–1. Samples were prepared as a film for liquid or neat for solid

substances. The intensities are given with vw (very weak, 120%), w (weak, 2040%), m

(medium, 4060%), s (strong, 6080%) and vs (very strong, 80100%) referring to the most

intense peak (100%).

8. LC/MS

LC/MS data was obtained on Agilent Technologies 1260 Infinity with C18 silica gel columns

and Agilent 1100 Series LC/MSD mass detector. As eluent, a mixture of MeCN (HPLC

grade) with H2O (Milli-Q Integral Water Purification System) and 0.1 vol-% formic acid as

additive was employed.





9. Additional Information: Aromatization upon Deprotection of

Protected Dimers 28 and S1

Dimers 28 and S1 were found to undergo aromatization by loss of formaldehyde upon

deprotection of the primary alcohol under a variety of conditions (Scheme S1). The different

reaction outcome compared to the parent cascade reaction could be rationalized by the fact

that dimers 28 and S1 have undergone tautomerization to a dienone system. This

tautomerization, which in the case of an unprotected primary alcohol seems to be slower than

the attack of the latter onto the carbonyl bridge, significantly increases the driving force for

aromatization. The orbital of the C1C2 σ-bond is parallel to the LUMO of the adjacent

enone system, which ensures a rapid retro-Aldol fragmentation of the molecule upon

deprotection of the primary alcohol S2 to catechol S3.

The resulting catechol S3 is unstable to purification, but was characterized by 1H NMR and

2D NMR spectroscopy and HRMS.

Scheme S1. Deprotection of protected dimers 28 and S1 resulting in aromatization by loss of formaldehyde.





10. Experimental procedures

2-methoxy-4-(methoxymethyl)phenol (S4)

Vanillyl alcohol 16 (7.00 g, 45.4 mmol) was dissolved in MeOH (70 ml), treated with

p-toluenesulfonic acid (432 mg, 2.27 mg, 0.05 eq.) and the mixture was stirred at rt for 7 h.

The mixture was then treated with NaHCO3 (210 mg), stirred for 15 min and then

concentrated under reduced pressure. The residue was taken up in EtOAc and eluted through a

pad of silica gel with EtOAc (total volume: 1 l). The collected elute was evaporated to give

the title compound S4 as a colorless oil (7.63 g, quant.).

TLC Rf = 0.51 (50% EtOAc/hexanes). 1H NMR (400 MHz, CHCl3): δ 6.89 – 6.80 (m, 3H), 5.62 (s, 1H), 4.38 (s, 2H), 3.90 (s, 3H),

3.37 (s, 3H) ppm. 13C NMR (100 MHz, CHCl3): δ 146.5, 145.2, 130.0, 121.1, 114.0, 110.4, 74.7, 57.8, 55.8 ppm.

HRMS ((–)-ESI, m/z): calc. [M–H–]: 167.07137; found: 167.07146 [M–H–].

IR 𝜈𝜈 = 3376 (br, w), 2935 (w), 1604 (w), 1514 (vs), 1451 (m), 1429 (s), 1362 (m), 1271

(vs), 1238 (vs), 1185 (vs), 1152 (vs), 1121 (s), 1079 (vs), 1031 (vs), 938 (w), 899 (m),

854 (m), 818 (s), 796 (s), 741 (s), 720 (m) cm–1.

4-hydroxy-3-methoxy-2-methylbenzaldehyde (S5)

A solution of n-BuLi in hexanes (c = 2.5 M, 74.0 ml, 185 mmol, 3.0 eq.) was added dropwise

to a solution of phenol S4 (10.35 g, 61.51 mmol) in THF (100 ml) at 10 °C. The resulting

mixture was stirred at 5 °C for 5 h and then cooled to 78 °C. MeI (13.4 ml, 30.6 g, 215

mmol, 3.5 eq.) was subsequently added very slowly and the resulting reaction mixture was

allowed to warm to 0 °C. Stirring was continued for 30 min before H2O (100 ml) and CH2Cl2





(300 ml) were added. The organic phase was washed with sat. aq. NaHCO3 (3x150 ml) and

brine (200 ml) and concentrated under reduced pressure.

The crude product was dissolved in CH2Cl2/H2O (95:5, 385 ml), the reaction vessel placed in

a water bath at 15 °C and DDQ (14.1 g, 62.1 mmol, 1.01 eq.) was added in two portions. The

water bath was removed and the reaction mixture stirred for 5 h before concentration under

reduced pressure. The suspension was filtered over a silica plug (3x6.5 cm, CH2Cl2) and the

crude product was purified by flash column chromatography (16x10 cm, 20304050%

Et2O/hexanes). The resulting mixed fractions were again subjected to flash column

chromatography to afford the title compound as a colorless solid (3.60 g, 35%).

TLC Rf = 0.24 (15% EtOAc/hexanes). 1H NMR (300 MHz, CDCl3): δ 10.05 (s, 1H), 7.54 (d, J = 8.4 Hz, 1H), 6.95 (d, J = 8.4 Hz,

1H), 6.24 (s, 1H), 3.81 (s, 3H), 2.61 (s, 3H) ppm. 13C NMR (75 MHz, CDCl3): δ 191.6, 154.1, 146.0, 134.3, 131.4, 128.6, 113.2, 61.4, 12.1 ppm.

MS (EI, %): 166.01 (100, M+), 151.00 (57), 123.01 (36), 77.02 (20).

HRMS (EI, m/z): calc. [M+]: 166.0630; found: 166.0621 [M+].

IR 𝜈𝜈 = 3239 (br, w), 2996 (vw), 2976 (vw), 2948 (vw), 2866 (vw), 2833 (vw), 2766 (vw),

1669 (s), 1585 (vs), 1491 (w), 1458 (vs), 1438 (w), 1410 (m), 1378 (vw), 1351 (vw)

1310 (vs), 1264 (w), 1212 (m), 1173 (vs), 1155 (vs), 1093 (m), 1031 (vw), 986 (m),

955 (vw), 879 (vw), 835 (vw), 821 (w), 783 (m), 764 (vw), 700 (vw), 664 (m) cm–1.

5-bromo-4-hydroxy-3-methoxy-2-methylbenzaldehyde (17)

Bromine (1.5 ml, 4.5 g, 28 mmol, 1.2 eq.) was added to a solution of aldehyde S5 (3.91 g,

23.5 mmol) and NaOAc (2.32 g, 28.3 mmol, 1.2 eq.) in HOAc (66 ml) at 15 °C. The cold

water bath was removed and the resulting solution was stirred at rt for 8.5 h. The reaction

mixture was poured onto ice water (290 ml) and subsequently filtered at 0 °C (Buchner funnel

sintered glass por. 4) to obtain the title compound as a colorless solid (5.59 g, 97%) after

drying under HV.

TLC Rf = 0.24 (20% EtOAc/hexanes).

m.p.: 157–160 °C.





1H NMR (600 MHz, CDCl3): δ 10.04 (s, 1H), 7.76 (s, 1H), 6.36 (s, 1H), 3.84 (s, 3H), 2.57 (s,

3H) ppm. 13C NMR (150 MHz, CDCl3): δ 190.2, 151.4, 146.5, 133.9, 132.9, 129.0, 107.0, 61.3, 11.8 ppm.

MS (EI, %): 243.93 (100, M+), 228.90 (64), 200.90 (32), 94.01 (22), 65.00 (35) Mass data

only given for 79Br containing compound.


IR 𝜈𝜈 = 3131 (br, w), 3067 (w), 2992 (w), 2944 (w), 2881 (vw), 2794 (vw), 2639 (vw),

1657 (s), 1561 (s), 1505 (vw), 1456 (m), 1418 (s), 1377 (vw), 1300 (vs), 1248 (m),

1203 (vs), 1164 (vs), 1076 (m), 989 (vs), 894 (m), 827 (s), 781 (vw), 737 (w), 703

(m), 688 (w) cm–1.

4,5-dihydroxy-3-methoxy-2-methylbenzaldehyde (S6)

Small scale preparation:

Bromide 17 (310 mg, 1.26 mmol) and NaOH (506 mg, 12.6 mmol, 10.0 eq.) were dissolved

in H2O (6.5 ml) and Cu powder (particle size 45 m, 4 mg, 0.06 mmol, 0.05 eq.) was added.

The resulting suspension was heated to reflux for 17 h, filtered and the pH of the resulting

solution was adjusted to pH = 7 with an aq. HCl solution (2 M). The aqueous phase was

extracted with EtOAc (3x15 ml), the combined organic phases washed with aq. sat.

Na2H2EDTA solution (50 ml), dried over Na2SO4 and concentrated under reduced pressure.

The crude product was purified by flash column chromatography (12x2.5 cm, 50%

EtOAc/hexanes) to afford the title compound as a brownish solid (173 mg, 75%).

The purity of the starting material proved to be crucial for the success of this reaction. Traces

of residual bromine led to failure presumably due to oxidation of the copper catalyst.

Reactions on larger scale were thwarted by a lack of reproducibility, lower conversion and

therefore lower isolated yield of the product. A homogeneous copper catalyst proved

beneficial (vide infra).

Large scale preparation:





CuSO4·5 H2O (70 mg, 0.28 mmol, 0.02 eq.) was added to a solution of NaOH (5.50 g, 138

mmol, 10.0 eq.) in deaerated H2O (55 ml) and the mixture was deaerated by passing a

nitrogen stream through the solution for 30 min. The solution was then cannulated onto

bromide 17 (4.94 g, 20.2 mmol) and the reaction mixture was heated to reflux for 9.5 h. After

cooling to rt, the suspension was acidified with aq. HCl solution (c = 6 M) and then filtered

through celite. The aqueous phase was extracted with EtOAc (3x200 ml), using addition of

MeOH (10 ml) to break the emulsion. The combined organic phases were dried over Na2SO4

and concentrated under reduced pressure. The crude residue was purified by flash column

chromatography on reverse-phase silica (10–20% MeCN/H2O + 0.1% FA) to give the title

compound S6 as an ocher solid (1.48 g, 59%).


m.p.: 148–150 °C. 1H NMR (400 MHz, D3COD): δ 10.01 (s, 1H), 7.09 (s, 1H), 3.76 (s, 3H), 2.48 (s, 3H) ppm. 13C NMR (100 MHz, D3COD): δ 192.9, 147.9, 146.5, 145.5, 128.3, 127.2, 114.7, 60.8, 10.7

ppm.

MS (EI, %): 182.96 (100, M+), 167.95 (31), 139.98 (36), 65.00 (30).


IR 𝜈𝜈 = 3308 (br, m), 2940 (w), 2849 (vw), 2730 (vw), 1665 (s), 1587 (s), 1495 (m), 1465

(s), 1429 (w), 1411 (w), 1374 (m), 1366 (m), 1315 (vs), 1226 (s), 1194 (m), 1105 (vs),

1017 (w), 946 (w), 874 (vw), 761 (vw), 731 (w), 705 (w) cm–1.

5-(hydroxymethyl)-3-methoxy-4-methylbenzene-1,2-diol (18)





Aldehyde S6 (1.17 g, 6.42 mmol) in THF (50 ml) at 0 °C was treated with water (1.17 ml)

followed by portion-wise addition of NaBH4 (243 mg, 6.42 mmol, 1.0 eq.) over 10 min. After

stirring for additional 5 min, the mixture was slowly treated with pH 5 phosphate buffer (c = 1

M, 40 ml) at 0 °C. The aqueous phase was extracted with EtOAc (150 ml) and the organic

phase washed with pH 5 buffer (c = 1 M, 2x50 ml) and brine (2x50 ml). The combined

aqueous layers were extracted with EtOAc (3x50 ml) and the resulting combined organic

phases dried over Na2SO4 and filtered. This solution was treated with silica gel (8 g) and

evaporated to dryness. (Note that concentration of the workup solution without silica gel

results in decomposition of the material). The silica-adsorbed product was purified flash

column chromatography on silica gel (15x3 cm, 66% EtOAc/hexanes) to afford the title

compound as a colorless solid (930 mg, 79%).

TLC Rf = 0.61 (100% EtOAc).

m.p.: 159160 °C (decomposition). 1H NMR (400 MHz, (D3C)2CO): δ 7.55 (s, 1H), 7.41 (s, 1H), 6.69 (s, 1H), 4.47 (d, J = 5.8 Hz,

2H), 3.83 (t, J = 5.8 Hz, 1H), 3.71 (s, 3H), 2.12 (s, 3H) ppm. 13C NMR (100 MHz, (D3C)2CO): δ 147.3, 144.1, 137.6, 132.0, 120.4, 111.6, 63.0, 60.3, 10.8

ppm.


IR 𝜈𝜈 = 3344 (br, w), 3202 (br, m), 2993 (w), 2936 (w), 2903 (w), 2832 (vw), 1619 (w),

1518 (w), 1489 (w), 1461 (m), 1431 (m), 1408 (w), 1379 (w), 1306 (s), 1260 (m),

1235 (s), 1216 (s), 1184 (m), 1093 (vs), 998 (s), 960 (s), 942 (vs), 879 (s), 864 (s), 832

(m), 759 (m), 713 (s), 692 (s), 642 (m), 620 (s), 579 (s), 563 (s) cm–1.

Epicolactone methyl ether 19 and regioisomer 20.

Benzyl alcohol 18 (150 mg, 0.814 mmol, 1.0 eq.) was suspended in MeCN (45 ml) and water

(90 ml). To this mixture was quickly added a solution of potassium ferricyanide (1.61 g,

4.89 mmol, 6.0 eq.) and NaHCO3 (410 mg, 4.89 mmol, 6.0 eq.) in water (15 ml).





Subsequently, epicoccine 10 (148 mg, 0.814 mmol) in MeCN/H2O (5:1, 9.0 ml) was added

via a syringe pump over 15 min at rt. After stirring for 1 h, pH 5 phosphate buffer (c = 1 M,

60 ml) was added and the aqueous phase extracted with EtOAc (4x100 ml). The combined

organic phases were washed with pH 5 phosphate buffer (c = 1 M, 50 ml) and brine (50 ml),

dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified

by flash column chromatography on silica gel (10x2.5 cm, 50% EtOAc/hexanes) to afford the

title compound as a colorless solid (124 mg, 42%). Further increasing the polarity of the

eluent during flash chromatography (66% EtOAc/hexanes) allows isolation of the regioisomer

20 as a colorless solid (46 mg, 16%). X-Ray suitable crystals of each compound were

obtained by slow diffusion of hexanes into a solution of the respective compound in EtOAc.

Epicolactone methyl ether 19

TLC Rf = 0.49 (5% MeOH/CH2Cl2).

m.p.: 250–260 °C (gradual decomposition) 1H NMR (800 MHz, [D6]DMSO): δ 8.51 (s, 1H), 6.00 (s, 1H), 4.53 (d, J = 9.7 Hz, 1H), 4.17

(d, J = 9.7 Hz, 1H), 3.91 (d, J = 9.7 Hz, 1H), 3.76 (d, J = 10.8 Hz, 1H), 3.66 (d,

J = 9.7 Hz, 1H), 3.57 (d, J = 10.8 Hz, 1H), 3.57 (s, 3H), 3.15 (s, 1H), 2.05 (s, 3H),

1.79 (s, 3H) ppm. 13C NMR (200 MHz, [D6]DMSO): δ 192.7, 189.9, 175.6, 149.2, 146.7, 144.3, 127.3, 91.0, 72.9,

70.1, 68.0, 67.9, 67.1, 66.5, 59.2, 49.8, 14.3, 13.6 ppm.


IR 𝜈𝜈 = 3423 (br, w), 2986 (vw), 2936 (vw), 2876 (vw), 1770 (s), 1729 (m), 1671 (vs),

1638 (s), 1475 (w), 1446 (w), 1392 (m), 1374 (m), 1356 (m), 1321 (w), 1289 (w),

1242 (s), 1218 (s), 1175 (s), 1145 (m), 1101 (s), 1081 (s), 1044 (vs), 1020 (m), 1003

(w), 982 (m), 967 (w), 952 (w), 933 (m), 885 (vw), 858 (vw), 803 (w), 789 (w), 744

(vw), 717 (w), 689 (w), 625 (vw) cm–1.





Regioisomer 20

TLC Rf = 0.40 (5% MeOH/CH2Cl2).

m.p.: 204–207 °C (decomposition) 1H NMR (400 MHz, [D6]DMSO): δ 9.00 (s, 1H), 6.29 (s, 1H), 4.64 (d, J = 10.2 Hz, 1H), 4.38

(d, J = 16.1 Hz, 1H), 4.27 (d, J = 16.1 Hz, 1H), 4.22 (d, J = 10.2 Hz, 1H), 3.85 (d,

J = 10.1 Hz, 1H), 3.75 (d, J = 10.1 Hz, 1H), 3.58 (s, 3H), 3.17 (s, 1H), 2.04 (s, 3H),


67.8, 66.3, 65.6, 59.8, 59.6, 59.5, 51.2, 17.4, 14.4 ppm.


IR 𝜈𝜈 = 3388 (w), 2991 (vw), 2929 (w), 2848 (vw), 1760 (m), 1695 (w), 1666 (vs), 1611

(w), 1483 (vw), 1449 (w), 1376 (m), 1324 (w), 1273 (w), 1229 (w), 1211 (w), 1180

(w), 1157 (w), 1143 (w), 1086 (s), 1042 (m), 1024 (m), 1014 (m), 976 (w), 915 (vw),

902 (vw), 857 (vw), 821 (vw), 744 (vw), 626 (vw) cm–1.

Epicolactone 1

Me-epicolactone 19 (29 mg, 0.080 mmol) was dried by co-evaporation from toluene and then

dissolved in THF (1.5 ml). The solution was treated with freshly distilled quinoline (38 μl,

0.32 mmol, 4.0 eq.) followed by MgI2 (334 mg, 1.20 mmol, 15.0 eq.). The suspension was

heated to 60 °C and stirred for 17 h. The mixture was cooled to rt, diluted with EtOAc (100

ml) and washed with an aq. HCl solution (c = 1 M, 3x30 ml) and brine (30 ml). The aqueous

layers were extracted with EtOAc (2x30 ml). The combined organic extracts were dried over





Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash

column chromatography on silica gel (12x1.3 cm, 2-2.5% MeOH/CH2Cl2) to afford the

natural product epicolactone (1) as a colorless solid (21.0 mg, 75%) as well as recovered

starting methyl ether 19 (4.5 mg, 16%) resulting in 89% yield based on recovered starting

material (brsm).

TLC Rf = 0.37 (5% MeOH/CH2Cl2). 1H NMR (400 MHz, [D6]DMSO): δ 8.63 (s, 1H), 8.52 (s, 1H), 6.08 (s, 1H), 4.51 (d, J = 9.6

Hz, 1H), 4.16 (d, J = 9.6 Hz, 1H), 3.91 (d, J = 9.6 Hz, 1H), 3.70 (d, J = 10.4 Hz, 1H),

3.65 (d, J = 9.6 Hz, 1H), 3.52 (d, J = 10.4 Hz, 1H), 3.06 (s, 1H), 1.97 (s, 3H), 1.79 (s,

3H) ppm. 13C NMR (100 MHz, [D6]DMSO): δ 192.8, 190.1, 175.9, 146.6, 145.8, 128.7, 127.4, 90.8, 73.0,

71.2, 68.3, 67.7, 67.2, 66.6, 50.1, 14.3, 13.0 ppm.


IR 𝜈𝜈 = 3410 (br, w), 2982 (vw), 2923 (w), 2852 (vw), 1769 (m), 1728 (m), 1670 (s), 1638

(s), 1473 (vw), 1445 (w), 1392 (m), 1375 (m), 1354 (s), 1241 (vs), 1218 (s), 1177 (s),

1143 (m), 1100 (s), 1076 (s), 1042 (vs), 1000 (m), 962 (w), 933 (m), 900 (w), 857 (w),

802 (w), 787 (w), 755 (w), 716 (w), 689 (w) cm–1.





1H NMR ([D6]DMSO):

chemical shift

δ/ppm (natural sample)1

chemical shift

δ/ppm (synthetic sample)

Δδ/ppm

8.61 (s, 1H) 8.63 (s, 1H) +0.02

8.50 (s, 1H) 8.52 (s, 1H) +0.02

6.07 (s, 1H) 6.08 (s, 1H) +0.01

4.51 (d, J = 9.6 Hz, 1H) 4.51 (d, J = 9.6 Hz, 1H) 0.00

4.16 (d, J = 9.6 Hz, 1H) 4.16 (d, J = 9.6 Hz, 1H) 0.00

3.90 (d, J = 9.8 Hz, 1H) 3.91 (d, J = 9.6 Hz, 1H) +0.01

3.70 (d, J = 10.4 Hz, 1H) 3.70 (d, J = 10.4 Hz, 1H) 0.00

3.65 (d, J = 9.8 Hz, 1H) 3.65 (d, J = 9.6 Hz, 1H) 0.00

3.53 (d, J = 10.4 Hz, 1H) 3.52 (d, J = 10.4 Hz, 1H) –0.01

3.06 (s, 1H) 3.06 (s, 1H) 0.00

1.96 (s, 3H) 1.97 (s, 3H) +0.01

1.79 (s, 3H) 1.79 (s, 3H) 0.00

1 da Silva Araújo, F. D.; de Lima Fávaro, L. C.; Araújo, W. L.; de Oliveira, F. L.; Aparicio, R.; Marsaioli, A. J.

Epicolactone – Natural Product Isolated from the Sugarcane Endophytic Fungus Epicoccum nigrum. Eur. J. Org. Chem. 2012, 5225 – 5230 (2012).





13C NMR ([D6]DMSO):

chemical shift

δ/ppm (natural sample)2

chemical shift

δ/ppm (synthetic sample)

Δδ/ppm

192.8 192.8 0.0

190.1 190.1 0.0

176.0 175.9 –0.1

146.6 146.6 0.0

145.9 145.8 –0.1

128.7 128.7 0.0

127.5 127.4 –0.1

90.9 90.8 –0.1

73.0 73.0 0.0

71.3 71.2 –0.1

68.3 68.3 0.0

67.7 67.7 0.0

67.2 67.2 0.0

66.6 66.6 0.0

50.1 50.1 0.0

14.4 14.3 –0.1

13.0 13.0 0.0

2 da Silva Araújo, F. D.; de Lima Fávaro, L. C.; Araújo, W. L.; de Oliveira, F. L.; Aparicio, R.; Marsaioli, A. J.

Epicolactone – Natural Product Isolated from the Sugarcane Endophytic Fungus Epicoccum nigrum. Eur. J. Org. Chem. 2012, 5225 – 5230 (2012).





isoepicolactone 21

regioisomer 20 (25 mg, 0.069 mmol) was dried by co-evaporation from toluene and then

dissolved in CH2Cl2 (6.0 ml). The solution was treated with dry AlCl3 (138 mg, 1.04 mmol,

15.0 eq.) and stirred vigorously at r.t. for 6.5 h. The mixture was then treated with aq.

Rochelle salt soln. (20 ml) and extracted with EtOAc (4x30 ml) using MeOH to break the

emulsion (4x1 ml). The combined organics were washed with brine (20 ml), dried over

Na2SO4 and concentrated under reduced pressure. The crude product was purified by filtration

through a short plug of silica gel, eluting with EtOAc to afford isoepicolactone (21) as a

colorless solid (23 mg, 96%).

TLC Rf = 0.15 (streak from baseline) (5% MeOH/CH2Cl2). 1H NMR (800 MHz, [D6]DMSO): δ 8.93 (s, 1H), 8.80 (s, 1H), 6.24 (s, 1H), 4.61 (d, J = 10.1

Hz, 1H), 4.38 (d, J = 16.1 Hz, 1H), 4.29 (d, J = 16.1 Hz, 1H), 4.22 (d, J = 10.1 Hz,

1H), 3.82 (d, J = 10.0 Hz, 1H), 3.75 (d, J = 10.0 Hz, 1H), 3.08 (s, 1H), 1.96 (s, 3H),


67.8, 66.4, 65.6, 60.0, 59.6, 51.3, 17.5, 13.8 ppm.


IR 𝜈𝜈 = 3398 (br, m), 2922 (w), 2851 (vw), 1769 (s), 1667 (vs), 1451 (w), 1382 (s), 1249

(m), 1229 (m), 1188 (m), 1156 (m), 1141 (m), 1081 (s), 1062 (m), 1040 (m), 1023

(m), 963 (vw), 915 (w), 803 (vw), 742 (vw), 696 (vw) cm–1.





The synthesis of protected analogs of alcohol 18 was carried out as follows:

4,5-bis(benzyloxy)-3-methoxy-2-methylbenzaldehyde (S7)

CuSO4·5 H2O (100 mg, 0.400 mmol, 0.02 eq.) was added to a solution of NaOH (8.06 g, 202

mmol, 10.0 eq.) in deaerated H2O (81 ml) and the mixture was deaerated by passing a

nitrogen stream through the solution for 30 min. The solution was gently heated to dissolve

the copper salt and then cannulated onto bromide 17 (4.94 g, 20.2 mmol). The reaction

mixture was heated to reflux for 16 h. After cooling to rt, the suspension was filtered and

acidified with aq. HCl solution (c = 6 M). The aqueous phase was extracted with EtOAc

(3x200 ml). The combined organic phases were dried over Na2SO4 and concentrated under

reduced pressure. The crude product was dissolved in acetone (100 ml) and K2CO3 (5.86 g,

42.4 mmol, 2.1 eq.) and BnBr (5.03 ml, 7.25 g, 42.4 mmol, 2.1 eq.) were added. The reaction

mixture was heated to reflux for 15 h and then cooled to rt. NEt3 (1.0 ml) was added and the

reaction mixture concentrated under reduced pressure. The residue was dissolved in EtOAc

(100 ml), washed with aq. HCl solution (c = 1 M, 2x100 ml), dried over Na2SO4 and

concentrated under reduced pressure. Purification of the crude product by flash column

chromatography (5–9% EtOAc/hexanes) to afford the compound S7 as a colorless oil (5.41 g,

74%).





TLC Rf = 0.31 (10% EtOAc/hexanes). 1H NMR (300 MHz, CDCl3): δ 10.24 (s, 1H), 7.47–7.29 (m, 11H), 5.15 (s, 2H), 5.13 (s, 2H),

3.86 (s, 3H), 2.54 (s, 3H) ppm. 13C NMR (75 MHz, CDCl3): δ 190.9, 152.9, 151.1, 147.3, 137.4, 136.6, 129.9, 129.1, 128.7,

128.5, 128.5, 128.2, 127.7, 110.4, 75.4, 71.2, 61.1, 10.4 ppm. 1 aromatic C missing

(overlapping)

MS (EI, %): 362.21 (2, M+), 271.12 (8), 181.09 (12), 91.04 (100), 65.02 (6).


IR 𝜈𝜈 = 3091 (vw), 3063 (vw), 3030 (vw), 2934 (vw), 2873 (vw), 2726 (vw), 1740 (vw),

1680 (s), 1589 (m), 1568 (w), 1497 (w), 1481 (m), 1452 (s), 1412 (w), 1376 (m), 1326

(vs), 1282 (s), 1240 (w), 1219 (w), 1199 (w), 1188 (w), 1118 (vs), 1079 (m), 1040 (s),

1028 (s), 1001 (m), 973 (m), 933 (w), 910 (w), 852 (w), 839 (w), 775 (w), 736 (s), 695

(vs) cm–1.

(4,5-bis(benzyloxy)-3-methoxy-2-methylphenyl)methanol (S8)

Benzaldehyde S8 (5.41 g, 14.9 mmol) was dissolved in EtOH (80 ml) and the solution cooled

to 0 °C. NaBH4 (845 mg, 22.4 mmol, 1.5 eq.) was introduced in 3 portions and the reaction

mixture was stirred at 0 °C for 2 h. The reaction mixture was treated with aq. phosphate

buffer (c = 1 M, pH 5, 150 ml) and the aqueous phase was extracted with EtOAc (2x200 ml).

The combined organic phases were washed with H2O (200 ml) and brine (400 ml), dried over

Na2SO4 and concentrated under reduced pressure. The product can be taken forward without

further purification.

The crude product can also be purified by flash column chromatography (12x4.5 cm, 1–1.5%

MeOH/CH2Cl2) on 10.4 mmol scale to yield the title compound as a pale yellow solid (3.19 g,

84% over 2 steps from S7).


m.p.: 5052 °C.





1H NMR (600 MHz, CDCl3): δ 7.48–7.45 (m, 2H), 7.45–7.42 (m, 2H), 7.39–7.35 (m, 2H),

7.35–7.29 (m, 4H), 5.09 (s, 2H), 5.04 (s, 2H), 4.64 (s, 2H), 3.86 (s, 3H), 2.21 (s, 3H),

1.56 (brs, 1H) ppm. 13C NMR (150 MHz, CDCl3): δ 152.7, 150.8, 141.6, 138.0, 137.3, 134.6, 128.6, 128.5, 128.4,

128.0, 127.6, 122.7, 109.5, 75.5, 71.3, 63.7, 61.0, 11.0 ppm. 1 aromatic C missing

(overlapping).

MS (EI, %): 364.15 (3, M+), 273.08 (8), 227.06 (3), 195.04 (3), 91.03 (100).


IR 𝜈𝜈 = 3396 (br, vw), 3089 (vw), 3063 (vw), 3031 (vw), 2921 (w), 2850 (w), 1658 (vw),

1594 (vw), 1497 (w), 1487 (m), 1452 (s), 1411 (m), 1372 (m), 1324 (s), 1278 (w),

1223 (w), 1187 (w), 1113 (vs), 1079 (m), 1035 (s), 1027 (s), 1002 (s), 974 (s), 929

(w), 910 (w), 880 (w), 840 (w), 773 (vw), 734 (s), 694 (vs), 592 (w) cm–1.

4,5-dihydroxy-3-methoxy-2-methylbenzyl acetate (26)

Benzyl alcohol S8 (1.0 g, 2.7 mmol) was dissolved in pyridine (14 ml) and Ac2O (0.34 l,

0.37 g, 3.6 mmol, 1.3 eq.) was added. The reaction mixture was stirred at rt for 25 h before aq.

HCl solution (2 M, 30 ml) was added. The aqueous phase was extracted with EtOAc (2x100

ml), the combined organic phases were washed with aq. HCl solution (2 M, 2x100 ml), dried

over Na2SO4 and concentrated under reduced pressure. The crude product S9 could be used

without further purification.

TLC Rf = 0.56 (20% EtOAc/hexanes) 1H NMR (400 MHz, CDCl3): δ 7.48–7.30 (m, 10H), 6.78 (s, 1H), 5.08 (s, 2H), 5.04 (s, 2H),

5.04 (s, 2H), 3.86 (s, 3H), 2.21 (s, 3H), 2.10 (s, 3H) ppm. 13C NMR (100 MHz, CDCl3): δ 171.1, 152.7, 150.7, 142.3, 137.9, 137.1, 129.6, 128.6, 128.5,

128.4, 128.0, 127.6, 124.2, 111.3, 75.4, 71.3, 65.0, 61.0, 21.2, 11.3 ppm. One C

missing (overlapping).

MS (EI, %): 406.14 (4, M+), 315.06 (4), 245.05 (7), 181.04 (2), 91.02 (100), 65.01 (4).






IR 𝜈𝜈 = 3031 (vw), 2933 (vw), 2865 (vw), 1735 (s), 1599 (vw), 1583 (vw), 1490 (m),

1453 (m), 1414 (w), 1372 (m), 1359 (m), 1328 (s), 1226 (vs), 1115 (vs), 1079 (w),

1027 (s), 957 (m), 930 (w), 911 (w), 879 (vw), 834 (w), 735 (s), 695 (vs) cm–1.

The crude product was dissolved in EtOAc (22 ml) and Pd/C (10 wt-%, 146 mg, 0.137 mmol,

0.05 eq.) was added. The reaction vessel was evacuated and filled with hydrogen gas for four

times and the reaction mixture was stirred under hydrogen gas atmosphere for 3.5 h at rt. The

reaction mixture was filtered over celite with EtOAc and the organic phase was concentrated

under reduced pressure to afford the title compound 26 as a pale green solid (623 mg, 100%

over 2 steps).


m.p.: 98–100 °C. 1H NMR (400 MHz, THF-d8): δ 7.73 (brs, 2H), 6.52 (s, 1H), 4.91 (s, 2H), 3.70 (s, 3H), 2.12 (s,

3H), 1.95 (s, 3H) ppm. 13C NMR (100 MHz, THF-d8): δ 170.4, 147.6, 144.9, 139.2, 125.9, 121.6, 113.6, 65.2, 60.1,

20.6, 11.1 ppm.

MS (EI, %): 226.03 (14, M+), 167.02 (36), 166.01 (100), 151.00 (28), 123.01 (26), 42.90

(40).


IR 𝜈𝜈 = 3390 (br, w), 2940 (vw), 2839 (vw), 1709 (s), 1606 (w), 1501 (m), 1466 (m), 1432

(w), 1363 (s), 1299 (s), 1217 (vs), 1090 (vs), 1018 (vs), 941 (m), 859 (w), 677 (w)

cm–1.

rac-((6S,10aS,10bR)-6,7-dihydroxy-9-methoxy-4,10-dimethyl-5,8,11-trioxo-3,5,6,8-

tetrahydro-1H,10aH-6,10b-methanobenzo[3,4]cyclohepta[1,2-c]furan-10a-yl)methyl

acetate (28)

Catechol 26 (38 mg, 0.17 mmol) was dissolved in dioxane (0.5 ml) and treated with Na2SO4

(250 mg), followed by Ag2O (230 mg, 1.00 mmol, 6.0 eq.). The dark-red mixture was treated

with epicoccine 10 (31 mg, 0.17 mmol, 1.0 eq.) in small portions over 3.5 h. The crude





product was purified by flash column chromatography on reverse-phase silica (10–20–30–

40% MeCN/H2O + 0.1% FA). The product-containing fractions were extracted with EtOAc

(2x) and the combined organic phases were washed with brine and concentrated under

reduced pressure to afford the title compound 28 as a colorless solid (34 mg, 50%). X-Ray

suitable crystals were obtained by slow diffusion of hexanes into a solution of the title

compound in EtOAc.

LC/MS Rt = 2.738 min (10–90% MeCN/H2O + 0.1% FA, 7 min, 2 ml/min).

m.p.: 205–215 °C (gradual decomposition). 1H NMR (800 MHz, (D3C)2CO): δ 8.28 (s, 1H, OH, C14OH), 5.24 (s, 1H, OH, C12OH), 4.88 (d,

J = 17.5 Hz, 1H, C8H), 4.73–4.69 (m, 2H, C6H, C8H), 4.42 (d, J = 11.0 Hz, 1H, C6H),

4.39 (d, J = 11.1 Hz, 1H, C2H), 4.36 (d, J = 11.1 Hz, 1H, C2H), 3.68 (s, 3H, C22H3),

2.21 (s, 3H, C18H3), 1.92 (s, 3H, C21H3), 1.73 (s, 3H, C19H3) ppm. 13C NMR (200 MHz, (D3C)2CO): δ 197.0 (C4), 189.3 (C11), 178.1 (C15), 169.8 (C20), 159.6

(C9), 153.1 (C16), 149.1 (C14), 142.2 (C17), 129.2 (C10), 123.7 (C13), 89.9 (C12),

69.9 (C8), 68.1 (C2), 66.8 (C6), 65.4 (C5), 60.0 (C22), 55.0 (C1), 20.5 (C21), 12.3

(C19), 12.1 (C18) ppm.

MS (EI, %): 404.08 (6, M+), 334.11 (14), 275.13 (32), 193.12 (26), 181.09 (32), 153.13

(42), 123.14 (20), 69.12 (28), 60.13 (42), 45.12 (51), 43.14 (100).

HRMS (EI, m/z): calc. [M+]: 404.1107; found: 404.1101 [M+]

IR 𝜈𝜈 = 3369 (w), 2926 (w), 2849 (vw), 1782 (s), 1739 (s), 1694 (vs), 1652 (vs), 1604 (w),

1455 (w), 1378 (m), 1291 (s), 1224 (vs), 1149 (m), 1100 (vs), 1049 (s), 1002 (m), 961

(w), 942 (w), 909 (m), 879 (w), 802 (vw), 767 (vw), 729 (m), 687 (vw) cm-1.

5-(((tert-butyldimethylsilyl)oxy)methyl)-3-methoxy-4-methylbenzene-1,2-diol (S11)

Benzyl alcohol S8 (431 mg, 1.18 mmol) was dissolved in CH2Cl2 (2.1 ml) and the solution

was cooled to 0 °C. Imidazole (121 mg, 1.77 mmol, 1.5 eq.) and TBSCl (232 mg, 1.54 mmol,

1.3 eq.) were added and the reaction mixture was stirred for 2 h. H2O (10 ml) was added and

the aqueous phase was extracted with CH2Cl2 (3x20 ml). The combined organic phases were





washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude

product was used directly in the following deprotection to catechol S10.

An analytical sample was purified by flash column chromatography (5% EtOAc/hexanes) to

afford the title compound as a colorless oil.

TLC Rf = 0.69 (10% EtOAc/hexanes). 1H NMR (400 MHz, CD2Cl2): δ 7.49–7.27 (m, 10H), 6.91 (s, 1H), 5.10 (s, 2H), 5.02 (s, 2H),

4.63 (s, 2H), 3.82 (s, 3H), 2.10 (s, 3H), 0.95 (s, 9H), 0.10 (s, 6H) ppm. The compound

shows rotamers in the 1H NMR spectrum. 13C NMR (100 MHz, CD2Cl2): δ 152.6, 150.7, 140.9, 138.6, 137.9, 135.6, 128.8, 128.7, 128.6,

128.2, 128.1, 127.9, 121.7, 108.4, 75.5, 71.2, 63.5, 61.1, 26.1, 18.7, 10.6, 5.2 ppm.

HRMS ((+)-ESI, m/z): calc. [M+Na+]: 501.2432; found: 501.2427 [M+Na+].

IR 𝜈𝜈 = 3065 (vw), 3031 (vw), 2952 (w), 2927 (w), 2881 (vw), 2855 (w), 1600 (vw), 1585

(vw), 1497 (w), 1486 (w), 1471 (w), 1452 (m), 1413 (w), 1371 (m), 1326 (m), 1252

(m), 1223 (w), 1188 (vw), 1116 (vs), 1055 (s), 1027 (s), 1004 (m), 937 (w), 910 (w),

834 (vs), 814 (m), 774 (vs), 732 (s), 694 (vs), 677 (m) cm–1.

Crude TBS ether S10 was dissolved in EtOAc (9.5 ml) and Pd/C (10 wt-%, 63 mg, 0.059

mmol, 0.05 eq.) was added. The reaction vessel was evacuated and filled with hydrogen gas

three times repetitively and stirred for 1.5 h. The reaction mixture was filtered over silica, the

filter cake washed with EtOAc and the organic phase concentrated. Purification of the residue

by flash column chromatography (12x2.5 cm, 20% EtOAc/hexanes) afforded the title

compound as a colorless solid (333 mg, 94% over 2 steps).


m.p.: 117118 °C. 1H NMR (400 MHz, (D3C)2CO): δ 7.52 (brs, 1H), 7.44 (brs, 1H), 6.71 (s, 1H), 4.60 (s, 2H),

3.71 (s, 3H), 2.09 (s, 3H), 0.91 (s, 9H), 0.08 (s, 6H) ppm. 13C NMR (100 MHz, (D3C)2CO): δ 147.3, 144.2, 137.7, 131.1, 120.1, 111.1, 64.1, 60.4, 26.3,

18.9, 10.8, 5.1 ppm.

HRMS (()-ESI, m/z): calc. [MH]: 297.1522; found: 297.1524 [MH].

IR 𝜈𝜈 = 3458 (br, m), 3235 (br, m), 2950 (m), 2927 (m), 2876 (w), 2850 (m), 1605 (m),

1494 (s), 1470 (m), 1459 (m), 1383 (w), 1365 (w), 1309 (s), 1251 (m), 1217 (s), 1171

(s), 1129 (s), 1106 (s), 1056 (m), 1009 (m), 937 (m), 866 (s), 850 (s), 837 (vs), 778 (s)

cm–1.





5-(((tert-butyldimethylsilyl)oxy)methyl)-3-methoxy-4-methylcyclohexa-3,5-diene-1,2-

dione (S12)

Catechol S11 (150 mg, 0.503 mmol) was dissolved in Et2O (9.5 ml) and o-chloranil (148 mg,

0.602 mmol, 1.2 eq.) was added. The reaction mixture was stirred for 1 min at rt and then

cooled to 78 °C. After stirring for 1 h, the suspension was filtered at 78 °C with precooled

glassware and the filter cake was washed with 78 °C cold Et2O to afford the product as a

bronze solid (96 mg, 64%).

m.p.: 98100 °C 1H NMR (400 MHz, (D3C)2CO): δ 6.32 (t, J = 2.1 Hz, 1H), 4.67 (d, J = 2.1 Hz, 2H), 3.81 (s,

3H), 2.03 (s, 3H), 0.96 (s, 9H), 0.16 (s, 6H) ppm. 13C NMR (100 MHz, (D3C)2CO): δ 179.9, 177.4, 157.9, 151.3, 134.1, 121.1, 63.1, 60.4, 26.2,

18.8, 10.7, 5.3 ppm.

HRMS ((+)-ESI, m/z): calc. [M+Na+]: 319.1336; found: 319.1335 [M+Na+]

IR 𝜈𝜈 = 2948 (vw), 2926 (w), 2892 (vw), 2853 (w), 1675 (w), 1659 (m), 1621 (vw), 1603

(vw), 1563 (w), 1472 (w), 1460 (w), 1441 (w), 1396 (w), 1371 (vw), 1360 (vw), 1328

(m), 1299 (w), 1252 (m), 1227 (w), 1196 (w), 1177 (w), 1128 (m), 1049 (s), 1004 (m),

935 (w), 874 (m), 835 (vs), 816 (s), 792 (m), 776 (vs), 704 (w), 676 (w) cm–1

rac-(6S,10aS,10bR)-10a-(((tert-butyldimethylsilyl)oxy)methyl)-6,7-dihydroxy-9-methoxy-

4,10-dimethyl-1H,3H-6,10b-methanobenzo[3,4]cyclohepta[1,2-c]furan-5,8,11(6H,10aH)-

trione (S1)





Quinone S12 (49 mg, 0.17 mmol, 2.0 eq.) was dissolved in dioxane (0.1 ml) and epicoccine

10 (15 mg, 0.083 mmol) in dioxane (0.1 ml) was added at rt. The reaction mixture was stirred

for 12 h and then concentrated under reduced pressure. The crude product was purified by

HPLC (40–90% MeCN/H2O + 0.15% FA, 45 min, 20ml/min, Rt = 25.7 min) to afford the title

compound as a waxy solid (8 mg, 19%).

The title compound can also be synthesized using the procedure described for the preparation

of acetate 28.

TLC Rf = 0.34 (5% MeOH/CH2Cl2). 1H NMR (600 MHz, CDCl3): δ 6.45 (brs, 1H, C14OH), 4.69 (dd, J = 17.3, 1.5 Hz, 1H, C8H),

4.61 (dd, J = 17.3, 1.5 Hz, 1H, C8H), 4.58 (d, J = 10.7 Hz, 1H, C6H), 4.51 (d, J

= 10.7 Hz, 1H, C6H), 4.35 (brs, 1H, C12OH), 3.87 (d, J = 9.6 Hz, 1H, C2H), 3.75 (s,

3H, C24H3), 3.68 (d, J = 9.6 Hz, 1H, C2H), 2.18 (s, 3H, C8H3), 1.76 (s, 3H, C19H3),

0.83 (s, 9H, 3xC23H3), 0.04 (s, 3H, C20H3 or C21H3), 0.01 (s, 3H, C20H3 or C21H3) ppm. 13C NMR (150 MHz, CDCl3): δ 195.0 (C4), 188.5 (C11), 177.3 (C15), 159.9 (C9), 151.8 (C16),

146.8 (C14), 143.7 (C17), 128.0 (C10), 124.3 (C13), 88.8 (C12), 71.0 (C2), 69.6 (C8),

66.7 (C6), 64.1 (C5), 60.2 (C24), 56.8 (C1), 25.9 (3×C23), 18.5 (C22), 12.7 (C19),

12.4 (C18), –5.7 (C20/C21), –5.8 (C20/C21) ppm.


IR 𝜈𝜈 = 3410 (w), 2927 (s), 2856 (m), 1782 (s), 1738 (w), 1691 (s), 1644 (s), 1548 (vw),

1463 (w), 1452 (w), 1376 (w), 1290 (m), 1251 (s), 1154 (m), 1101 (vs), 1006 (w), 940

(w), 838 (vs), 782 (w), 699 (w) cm–1.

rac-(6S,10bS)-6,7,8-trihydroxy-9-methoxy-4,10-dimethyl-1H,3H-6,10b-

methanobenzo[3,4]cyclohepta[1,2-c]furan-5,11(6H)-dione (S3)

As a representative example for the decomposition of dimers 28 and S1, the following

procedure is provided:

TBS ether S1 was dissolved in CD2Cl2 (0.6 ml) and the solution treated with TFA-d1 (3

drops). The product formed instantaneously as judged by NMR analysis.





The described procedure represents one of many examples of the formation of catechol S3

upon deprotection of protected alcohols 28 or S1.

1H NMR (600 MHz, CD2Cl2): δ 5.03 (dq, J = 17.5, 1.4 Hz, 1H), 4.82 (d, J = 10.6 Hz, 1H), 4.79

(dq, J = 17.5, 1.5 Hz, 1H), 4.69 (d, J = 10.6 Hz, 1H), 3.85 (s, 3H), 2.34 (s, 3H), 1.76

(s, 3H) ppm.






11. X-Ray Single Crystal Structure

11.1 Epicolactone Me-Ether 19 CCDC 1062999

Crystallographic data.

net formula C18H18O8 Mr/g mol−1 362.32 crystal size/mm 0.100 × 0.100 × 0.090 T/K 173(2) Radiation MoKα Diffractometer 'Bruker D8Quest' crystal system monoclinic space group 'P 21/c' a/Å 13.2284(7) b/Å 7.8031(4) c/Å 15.7251(7) α/° 90 β/° 107.0479(13) γ/° 90 V/Å3 1551.86(13) Z 4





calc. density/g cm−3 1.551 μ/mm−1 0.123 absorption correction multi-scan transmission factor range 0.9290–0.9705 refls. Measured 26422 Rint 0.0375 mean σ(I)/I 0.0226 θ range 2.709–26.54 observed refls. 2660 x, y (weighting scheme) 0.0529, 0.6811 hydrogen refinement mixed refls in refinement 3183 Parameters 246 Restraints 0 R(Fobs) 0.0409 Rw(F2) 0.1027 S 1.060 shift/errormax 0.001 max electron density/e Å−3 0.257 min electron density/e Å−3 −0.296

C-H: constr, O-H: refall.

Number Label Charge SybylType Xfrac + ESD Yfrac + ESD Zfrac + ESD Symm. op. 1 C1 0 C.3 0.161479 0.703319 0.241212 x,y,z 2 C2 0 C.3 0.221495 0.872425 0.263456 x,y,z 3 H2 0 H 0.171641 0.971827 0.249341 x,y,z 4 C3 0 C.2 0.302282 0.884943 0.212699 x,y,z 5 C4 0 C.2 0.348802 0.725933 0.192366 x,y,z 6 C5 0 C.2 0.326987 0.574732 0.222981 x,y,z 7 C6 0 C.3 0.254293 0.574991 0.282394 x,y,z 8 C7 0 C.3 0.2152 0.402422 0.305548 x,y,z 9 H7A 0 H 0.153025 0.362658 0.257446 x,y,z

10 H7B 0 H 0.271553 0.314659 0.315879 x,y,z 11 C8 0 C.3 0.27259 0.535277 0.439871 x,y,z 12 H8A 0 H 0.332173 0.460131 0.471199 x,y,z 13 H8B 0 H 0.24927 0.600916 0.484626 x,y,z 14 C9 0 C.3 0.306151 0.65781 0.375881 x,y,z 15 C10 0 C.3 0.267843 0.848545 0.366433 x,y,z 16 C11 0 C.3 0.369593 0.950048 0.40943 x,y,z 17 H11A 0 H 0.373421 1.05244 0.373325 x,y,z 18 H11B 0 H 0.371508 0.98791 0.46997 x,y,z 19 C12 0 C.2 0.425312 0.671064 0.399132 x,y,z 20 C13 0 C.2 0.071799 0.699052 0.285079 x,y,z 21 C14 0 C.2 0.085278 0.802356 0.365723 x,y,z 22 C15 0 C.2 0.176832 0.881506 0.405198 x,y,z 23 C16 0 C.3 0.370018 0.410274 0.200677 x,y,z 24 H16A 0 H 0.311552 0.335972 0.168555 x,y,z 25 H16B 0 H 0.4108 0.353354 0.255571 x,y,z





26 H16C 0 H 0.416062 0.433106 0.163205 x,y,z 27 C17 0 C.3 -0.033741 0.681393 0.434323 x,y,z 28 H17A 0 H -0.108823 0.689181 0.430721 x,y,z 29 H17B 0 H 0.008775 0.678024 0.496884 x,y,z 30 H17C 0 H -0.021365 0.576838 0.404314 x,y,z 31 C18 0 C.3 0.189363 0.998526 0.482426 x,y,z 32 H18A 0 H 0.208607 1.11321 0.467032 x,y,z 33 H18B 0 H 0.24513 0.954799 0.533615 x,y,z 34 H18C 0 H 0.122598 1.00457 0.497449 x,y,z 35 O1 0 O.3 0.116474 0.679372 0.149122 x,y,z 36 H1 0 H 0.062789 0.608128 0.144215 x,y,z 37 O2 0 O.2 0.325543 1.02203 0.186516 x,y,z 38 O3 0 O.3 0.411849 0.741322 0.137372 x,y,z 39 H3 0 H 0.420393 0.849923 0.131536 x,y,z 40 O4 0 O.3 0.187435 0.43633 0.385323 x,y,z 41 O5 0 O.3 0.456688 0.835464 0.413273 x,y,z 42 O6 0 O.2 0.487026 0.556055 0.405508 x,y,z 43 O7 0 O.2 -0.010193 0.625511 0.247187 x,y,z 44 O8 0 O.3 -0.004133 0.828226 0.391875 x,y,z





11.2 Regioisomer 20 CCDC 1063000


net formula C18H18O8 Mr/g mol−1 362.32 crystal size/mm 0.090 × 0.080 × 0.020 T/K 100(2) Radiation MoKα Diffractometer 'Bruker D8Venture' crystal system monoclinic space group 'P 21/n' a/Å 8.9094(7) b/Å 13.7653(10) c/Å 13.1489(9) α/° 90 β/° 103.0714(19) γ/° 90 V/Å3 1570.8(2) Z 4 calc. density/g cm−3 1.532 μ/mm−1 0.122 absorption correction multi-scan transmission factor range 0.6891–0.7453 refls. Measured 18028 Rint 0.0406 mean σ(I)/I 0.0314





θ range 3.119–26.03 observed refls. 2431 x, y (weighting scheme) 0.0395, 0.8828 hydrogen refinement mixed refls in refinement 3088 Parameters 246 Restraints 0 R(Fobs) 0.0360 Rw(F2) 0.0881 S 1.022 shift/errormax 0.001 max electron density/e Å−3 0.323 min electron density/e Å−3 −0.217


Number Label Charge SybylType Xfrac + ESD Yfrac + ESD Zfrac + ESD Symm. op. 1 C1 0 C.3 0.788553 0.431642 0.079291 x,y,z 2 H1A 0 H 0.781662 0.444844 0.004362 x,y,z 3 H1B 0 H 0.878513 0.389088 0.105677 x,y,z 4 C2 0 C.2 0.803634 0.523968 0.139485 x,y,z 5 C3 0 C.2 0.912908 0.59204 0.150817 x,y,z 6 C4 0 C.2 0.895594 0.680869 0.208995 x,y,z 7 C5 0 C.3 0.751943 0.690269 0.252506 x,y,z 8 H5 0 H 0.728219 0.760058 0.263043 x,y,z 9 C6 0 C.3 0.613864 0.639498 0.182626 x,y,z

10 C7 0 C.3 0.673926 0.531521 0.196061 x,y,z 11 C8 0 C.3 0.561518 0.45379 0.141118 x,y,z 12 H8A 0 H 0.513506 0.419851 0.192088 x,y,z 13 H8B 0 H 0.478978 0.483839 0.087101 x,y,z 14 C9 0 C.2 0.469215 0.654629 0.225768 x,y,z 15 C10 0 C.2 0.490241 0.672253 0.33818 x,y,z 16 C11 0 C.2 0.630075 0.662562 0.40269 x,y,z 17 C12 0 C.3 0.762234 0.630959 0.354968 x,y,z 18 C13 0 C.3 0.922538 0.632842 0.427394 x,y,z 19 H13A 0 H 0.975895 0.694447 0.419161 x,y,z 20 H13B 0 H 0.915352 0.62659 0.501149 x,y,z 21 C14 0 C.2 0.906737 0.484602 0.341594 x,y,z 22 C15 0 C.3 0.741517 0.522526 0.317362 x,y,z 23 C16 0 C.3 0.313416 0.789638 0.369224 x,y,z 24 H16A 0 H 0.214565 0.795808 0.38952 x,y,z 25 H16B 0 H 0.302681 0.813546 0.297734 x,y,z 26 H16C 0 H 0.391772 0.827964 0.416874 x,y,z 27 C17 0 C.3 0.653194 0.67368 0.51836 x,y,z 28 H17A 0 H 0.728266 0.725429 0.542703 x,y,z 29 H17B 0 H 0.691572 0.612471 0.552652 x,y,z 30 H17C 0 H 0.55489 0.690426 0.535511 x,y,z 31 C18 0 C.3 0.651508 0.457147 0.376759 x,y,z 32 H18A 0 H 0.695736 0.462378 0.451894 x,y,z





33 H18B 0 H 0.657813 0.389608 0.354525 x,y,z 34 H18C 0 H 0.543389 0.477627 0.361859 x,y,z 35 O1 0 O.3 0.649303 0.386306 0.093856 x,y,z 36 O2 0 O.3 1.03787 0.58464 0.108518 x,y,z 37 H2 0 H 1.11196 0.628689 0.138892 x,y,z 38 O3 0 O.2 0.993945 0.744276 0.221607 x,y,z 39 O4 0 O.3 0.587094 0.671971 0.079235 x,y,z 40 H4 0 H 0.497422 0.654285 0.048511 x,y,z 41 O5 0 O.2 0.342786 0.651205 0.165238 x,y,z 42 O6 0 O.3 0.359237 0.689278 0.374421 x,y,z 43 O7 0 O.3 1.00521 0.550592 0.396933 x,y,z 44 O8 0 O.2 0.950873 0.406606 0.319577 x,y,z





11.3 rac-((6S,10aS,10bR)-6,7-dihydroxy-9-methoxy-4,10-dimethyl-5,8,11-trioxo-

3,5,6,8-tetrahydro-1H,10aH-6,10b-methanobenzo[3,4]cyclohepta[1,2-c]furan-10a-

yl)methyl acetate (28) CCDC 1063001


net formula C20H20O9 Mr/g mol−1 404.36 crystal size/mm 0.100 × 0.090 × 0.080 T/K 100(2) Radiation MoKα Diffractometer 'Bruker D8Venture' crystal system monoclinic space group 'P 21/n' a/Å 12.4719(4) b/Å 10.4198(3) c/Å 14.7987(5) α/° 90 β/° 109.9144(10) γ/° 90 V/Å3 1808.16(10) Z 4 calc. density/g cm−3 1.485 μ/mm−1 0.118 absorption correction multi-scan transmission factor range 0.9259–0.9585 refls. Measured 43959 Rint 0.0356





mean σ(I)/I 0.0172 θ range 2.442–26.43 observed refls. 3097 x, y (weighting scheme) 0.0453, 1.1557 hydrogen refinement mixed refls in refinement 3711 Parameters 274 Restraints 0 R(Fobs) 0.0379 Rw(F2) 0.1001 S 1.045 shift/errormax 0.001 max electron density/e Å−3 0.304 min electron density/e Å−3 −0.279


Number Label Charg

e SybylTyp

e Xfrac +

ESD Yfrac +

ESD Zfrac +

ESD Symm.

op.

1 C1 0 C.2 0.15802 0.48743 0.493802 x,y,z

2 C2 0 C.2 0.153128 0.494621 0.392519 x,y,z

3 C3 0 C.2 0.210428 0.38984 0.358674 x,y,z

4 C4 0 C.2 0.246097 0.285861 0.412974 x,y,z

5 C5 0 C.3 0.33136 0.188454 0.402608 x,y,z

6 C6 0 C.2 0.447135 0.261039 0.438561 x,y,z

7 C7 0 C.2 0.487265 0.298146 0.540839 x,y,z

8 C8 0 C.2 0.429617 0.254467 0.595986 x,y,z

9 C9 0 C.3 0.465468 0.263791 0.702898 x,y,z

10 H9A 0 H 0.544734 0.232795 0.733012 x,y,z

11 H9B 0 H 0.461505 0.35391 0.722715 x,y,z

12 C10 0 C.3 0.325027 0.104354 0.652495 x,y,z

13 H10A 0 H 0.246784 0.090534 0.653204 x,y,z

14 H10B 0 H 0.362879 0.01994 0.656799 x,y,z

15 C11 0 C.3 0.321749 0.174398 0.561068 x,y,z

16 C12 0 C.2 0.339183 0.090944 0.481755 x,y,z

17 C13 0 C.3 0.217459 0.261356 0.502784 x,y,z

18 C14 0 C.2 0.191456 0.382277 0.548677 x,y,z

19 C15 0 C.3 0.104398 0.185198 0.473909 x,y,z





20 H15A 0 H 0.085229 0.162839 0.531627 x,y,z

21 H15B 0 H 0.041332 0.236958 0.430312 x,y,z

22 C16 0 C.2 0.027344 0.000281 0.381469 x,y,z

23 C17 0 C.3 0.055898 -0.116356 0.336216 x,y,z

24 H17A 0 H -0.005507 -0.133853 0.275389 x,y,z

25 H17B 0 H 0.064602 -0.189773 0.379526 x,y,z

26 H17C 0 H 0.127403 -0.102248 0.323932 x,y,z

27 C18 0 C.3 0.204724 0.68852 0.569008 x,y,z

28 H18A 0 H 0.234582 0.721004 0.520111 x,y,z

29 H18B 0 H 0.266765 0.64914 0.621456 x,y,z

30 H18C 0 H 0.171999 0.759661 0.594311 x,y,z

31 C19 0 C.3 0.596101 0.372678 0.579197 x,y,z

32 H19A 0 H 0.607762 0.422259 0.527035 x,y,z

33 H19B 0 H 0.660022 0.313294 0.606015 x,y,z

34 H19C 0 H 0.591737 0.431252 0.62964 x,y,z

35 C20 0 C.3 0.183155 0.377959 0.647956 x,y,z

36 H20A 0 H 0.137586 0.45075 0.656202 x,y,z

37 H20B 0 H 0.259812 0.382767 0.696123 x,y,z

38 H20C 0 H 0.146673 0.297555 0.65605 x,y,z

39 O1 0 O.3 0.118077 0.594447 0.526755 x,y,z

40 O2 0 O.2 0.111882 0.586354 0.340765 x,y,z

41 O3 0 O.3 0.231081 0.405477 0.275456 x,y,z

42 H3 0 H 0.208852 0.478367 0.252091 x,y,z

43 O4 0 O.3 0.312335 0.137191 0.311612 x,y,z

44 H4 0 H 0.364803 0.170056 0.294332 x,y,z

45 O5 0 O.2 0.496971 0.281989 0.382747 x,y,z

46 O6 0 O.3 0.388741 0.185582 0.731374 x,y,z

47 O7 0 O.2 0.363312 -0.020623 0.483597 x,y,z

48 O8 0 O.3 0.121217 0.070796 0.426127 x,y,z

49 O9 0 O.2 -0.065754 0.031069 0.380747 x,y,z





12. NMR spectra

S4 (1H NMR, 400 MHz, CDCl3)

S4 (13C NMR, 100 MHz, CDCl3)











17 (1H NMR, 600 MHz, CDCl3)

17 (13C NMR, 150 MHz, CDCl3)





S6 (1H NMR, 400 MHz, D3COD)

S6 (13C NMR, 100 MHz, D3COD)





18 (1H NMR, 400 MHz, (D3C)2CO)

18 (13C NMR, 100 MHz, (D3C)2CO)





19 (1H NMR, 800 MHz, [D6]DMSO)

19 (13C NMR, 200 MHz, [D6]DMSO)





20 (1H NMR, 400 MHz, [D6]DMSO)

20 (13C NMR, 100 MHz, [D6]DMSO)





Epicolactone 1 (1H NMR, 400 MHz, [D6]DMSO)

Epicolactone 1 (13C NMR, 100 MHz, [D6]DMSO)





isoepicolactone 21 (1H NMR, 400 MHz, [D6]DMSO)

isoepicolactone 21 (13C NMR, 200 MHz, [D6]DMSO)























26 (1H NMR, 400 MHz, THF-d8)

26 (13C NMR, 100 MHz, THF-d8)





28 (1H NMR, 800 MHz, (D3C)2CO)

28 (1H NMR, 200 MHz, (D3C)2CO)





S10 (1H NMR, 400 MHz, CD2Cl2)

S10 (13C NMR, 100 MHz, CD2Cl2)





S11 (1H NMR, 400 MHz, (D3C)2CO)

S11 (13C NMR, 100 MHz, (D3C)2CO)





S12 (1H NMR, 400 MHz, (D3C)2CO)

S12 (13C NMR, 100 MHz, (D3C)2CO)








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SUEMENTARY INRMATIN - Nature Research · mixture was stirred at 5 °C for 5 h and then cooled to 78 °C. MeI (13.4 ml, 30.6 g, 215 mmol, 3.5 eq.) was subsequently added very slowly