supporting information · 2zn (1.2 m in toluene, 2.3 ml, 2.72 mmol, 1.5 eq) was added dropwise over...

Supporting Information

© Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2008

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Asymmetric Total Synthesis of PDIM A, a Virulence Factor of Mycobacterium

tuberculosis

Eva Casas-Arce, Bjorn ter Horst, Ben L. Feringa* and Adriaan Minnaard*

Stratingh Institute for Chemistry

University of Groningen, Nijenborgh 4, 9747 AG

Groningen (The Netherlands)

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General experimental remarks: All reactions were carried out under nitrogen atmosphere and in flame

dried glassware unless otherwise noted. Reagents were purchased from Aldrich, Acros Chimica, Merck or

Fluka and were used as received unless otherwise stated. All solvents were reagent grade and were dried

and distilled before use according to standard procedures. Column chromatography was performed using

pure silica gel (Merck type 9385, 230-400 mesh) as the stationary phase. TLC was performed using silica

gel plates (Merck type 60, 0.25 mm) and the components were visualized by staining with KMnO4 or

phosphomolybdic acid. CI-MS and EI-HRMS spectra were recorded on an AEI MS-902, ESI-HRMS

measurements were carried out using a LTQ-Orbitrap XL (Thermo Fischer Scientific) mass spectrometer

and MALDI-TOF spectra were recorded on a Voyager-OE.Pro (Applied Biosystems). 1H, 13C and 19F

NMR spectra were carried out in CDCl3 on a Varian AMX400 (100.59 MHz) spectrometer. Chemical

shift values are denoted in δ values (ppm) relative to residual solvent peaks (CHCl3, 1H δ = 7.26, 13C δ =

77.0). Multiplicity was determined by APT 13C experiments. Data are reported as follows: chemical shift,

multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, td = triple doublet and m = multiplet),

coupling constants and integration. Optical rotations were measured on a Schmidt + Haensch polarimeter

(Polartronic MH8) with a 10 cm cell (c given in g/100 ml). Melting points were determined on a Buchi

B–545 melting point apparatus. The enantiomeric excess of 17 was determined by GC analysis (CP

Chiralsil Dex CB column, 30 m x 0.25 mm) using a flame ionization detector (in comparison with

racemic product). Racemic 17 was prepared by reaction of cycloheptenone with MeMgBr at 0 ºC in THF

for 2 h in the presence of Cu(OTf)2.

OO

OP N

Ph

Ph

(S,R,R)-L*

(3S)-methylcycloheptanone (17): (S,R,R)-L*1 (9.75 mg, 0.018 mmol, 1.0 mol%) and Cu(OTf)2 (3.28 mg,

0.009 mmol, 0.5 mol%) were dissolved in dry toluene (4 ml) and stirred for 30 min under nitrogen at

room temperature. The mixture was cooled at -25 ºC and Me2Zn (1.2 M in toluene, 2.3 ml, 2.72 mmol,

1.5 eq) was added dropwise over 2 min. After stirring for 10 min, a solution of cycloheptenone (200 mg,

1.82 mmol) in dry toluene (4 ml) was added over 5 h by syringe pump and the resulting mixture was

stirred overnight at -25 ºC. The reaction was quenched with aq. NH4Cl (sat.), extracted with Et2O, washed

with brine (sat.) and dried (Na2SO4). The solvents were removed under reduced pressure and the product

1 For the synthesis of the lignad, see: Feringa, B. L.; Pineschi, M.; Arnold, L. A.; Imbos, R.; de Vries, A. H. M. Angew. Chem. Int. Ed. Engl. 1997, 36, 2620-2623.

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was purified by column chromatography (n-pentane-Et2O 50:1) to give (3S)-17 (213 mg, 1.69 mmol, 93%,

95% ee) as a colorless oil.

[α]D22 = – 46.1 º (c = 0.8, MeOH). Proof of stereochemistry: reported positive optical rotation is assigned

to the R enantiomer, [a]D25 = + 55.2 (MeOH).2 1H-NMR (CDCl3, 400 MHz) δ = 0.99 (d, J = 6.7 Hz, 3H),

1.29 (m, 1H), 1.40 (m, 1H), 1.61 (m, 2H), 1.84 (m, 3H), 2.41 (d, J = 7.3 Hz, 2H), 2.47 (t, J = 4.7 Hz, 2H)

ppm. Ee determination by chiral GC analysis, CP Chiralsil Dex CB column, T = 100 ºC, retention times:

10.13 (S) / 10.94 (R) min.

Methylcycloheptanone (racemic)

(3S)-methylcycloheptanone (17)

2 Lightner, D. A.; Docks, E. L.; Tetrahedron 1979, 35, 713-720.

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OO

OP N

Ph

Ph

(S,R,R)-L*

(2R)-ethyl-(3S)-methylcycloheptanone (6): (S,R,R)-L* (146 mg, 0.27 mmol, 1.0 mol%) and Cu(OTf)2

(49 mg, 0.14 mmol, 0.5 mol%) were dissolved in dry toluene (60 ml) and stirred for 30 min under

nitrogen at room temperature. The mixture was cooled at -25 ºC and Me2Zn (1.2 M in toluene, 34 ml,

40.36 mmol, 1.5 eq) was added dropwise over 5 min. After stirring for 10 min, a solution of

cycloheptenone (3.0 ml, 26.91 mmol) in dry toluene (60 ml) was added over 5 h by syringe pump and the

resulting mixture was stirred overnight at -25 ºC. Ethyl iodide (22 ml, 269 mmol, 10.0 eq) and

hexamethylphosphoramide (47 ml, 269 mmol, 10.0 eq) were added, the mixture was warmed up to 0 ºC

and stirred for 60 h. The reaction was quenched with aq. NH4Cl (sat.), extracted with Et2O, washed with

brine (sat.) and dried (Na2SO4). The solvents were removed under reduced pressure and the product was

purified by column chromatography (n-pentane-Et2O 50:1) to give 6 (3.44 g, 22.3 mmol, 83%, >95:5

trans:cis, 95% ee for trans) as a colorless oil.

[α]D22 = – 49.7 º (c = 4.9, CHCl3). 1H-NMR (CDCl3, 400 MHz) δ = 0.82 (t, J = 7.4 Hz, 3H), 0.99 (d, J =

6.7 Hz, 3H), 1.20 (m, 1H), 1.32-1.69 (m, 6H), 1.83 (m, 2H), 1.99 (td, J = 3.8 Hz, J = 10.9 Hz, 1H), 2.27

(m, 1H), 2.55 (td, J = 3.6 Hz, J = 11.8 Hz, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 11.9 (CH3), 21.0

(CH3), 24.1 (CH2), 26.0 (CH2), 28.0 (CH2), 35.6 (CH), 36.1 (CH2), 41.2 (CH2), 61.8 (CH), 216.0 (C) ppm.

MS (CI) for C10H18O: m/z = 172.2 [M+NH4]+, HRMS (EI) calcd for C10H18O: 154.1358, found: 154.1353.

OO

(8R)-ethyl-(7S)-methyloxocan-2-one (7): A solution of 6 (3.30 g, 21.39 mmol) in DCM (15 ml) was

added to a suspension of m-chloroperbenzoic acid (70-75%, 25.28 g, 106.95 mmol, 5.0 eq) in DCM (15

ml) and the resulting mixture was refluxed for 60 h. The reaction mixture was cooled to room temperature,

washed with aq. NaHCO3 (sat.) and aq. Na2S2O3 (sat.) and then dried (Na2SO4). The DCM was removed

under reduced pressure and the product was purified by column chromatography (n-pentane-Et2O 50:1) to

give 7 (2.18 g, 12.83 mmol, 60%) as a colorless oil. [α]D22 = – 54.0 º (c = 5.2, CHCl3). 1H-NMR (CDCl3,

400 MHz) δ = 0.85 (d, J = 6.9 Hz, 3H), 0.95 (t, J = 7.4 Hz, 3H), 1.28 (m, 1H), 1.41-1.92 (m, 8H), 2.38 (t,

J = 6.4 Hz, 2H), 4.28 (td, J = 2.5 Hz, J = 9.5 Hz, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 9.9 (CH3),

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17.8 (CH3), 24.3 (CH2), 25.8 (CH2), 29.6 (CH2), 33.4 (CH), 33.7 (CH2), 40.9 (CH2), 84.6 (CH), 177.3 (C)

ppm. MS (CI) for C10H18O2: m/z = 188.2 [M+NH4]+, HRMS (EI) calcd for C10H18O2-C2H5: 141.0915,

found: 141.0921.

MeO

O OH

(7R)-hydroxy-(6S)-methylnonanoic acid methyl ester (8): To a solution of 7 (1.81 g, 10.63 mmol) in

methanol (30 ml), activated K2CO3 (1.47 g, 10.63 mmol, 1.0 eq)3 was added and the resulting mixture

was stirred for 3 h at room temperature. The reaction was quenched with aq. NH4Cl (sat.) and the

methanol was removed under reduced pressure. The product was then extracted with DCM and the

solution was dried (Na2SO4). The solvent was removed under reduced pressure and the product was

purified by column chromatography (n-pentane-EtOAc 9:1) to give 8 (1.94 g, 9.57 mmol, 90%) as a

colorless oil. [α]D22 = – 9.9 º (c = 2.8, CHCl3). 1H-NMR (CDCl3, 400 MHz) δ = 0.85 (d, J = 6.8 Hz, 3H),

0.93 (t, J = 7.4 Hz, 3H), 1.03-1.68 (m, 9H, 1OH), 2.29 (t, J = 7.2 Hz, 2H), 3.30 (m, 1H), 3.63 (s, 3H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.3 (CH3), 15.3 (CH3), 25.2 (CH2), 26.2 (CH2), 26.7 (CH2), 31.4

(CH2), 34.0 (CH2), 38.2 (CH), 51.4 (CH3), 77.3 (CH), 174.2 (C) ppm. MS (CI) for C11H22O3: m/z = 220.3

[M+NH4]+, HRMS (EI) calcd for C11H22O3-C2H5: 173.1178, found: 173.1172.

MeO

O O

(7R)-methoxy-(6S)-methylnonanoic acid methyl ester (9): To a solution of 8 (1.00 g, 4.94 mmol) in

dry DMF (30 ml), MeI (4.62 ml, 74.15 mmol, 15.0 eq) and NaH (60%, 1.98 g, 49.43 mmol, 10.0 eq) were

added at 0 ºC. The resulting suspension was stirred overnight under nitrogen at 40 ºC, after which the

reaction was quenched with H2O and extracted with Et2O. The combined organic layers were washed

with brine (sat.), dried (MgSO4) and concentrated. The product was purified by column chromatography

(n-pentane-EtOAc 9:1) to give 9 (983 mg, 4.54 mmol, 92%) as a colorless oil. [α]D22 = – 3.1 º (c = 2.8,

CHCl3). 1H-NMR (CDCl3, 400 MHz) δ = 0.81 (d, J = 6.9 Hz, 3H), 0.89 (t, J = 7.4 Hz, 3H), 1.09 (m, 1H),

1.23 (m, 1H), 1.32-1.50 (m, 4H), 1.54-1.72 (m, 3H), 2.30 (t, J = 7.6 Hz, 2H), 2.84 (m, 1H), 3.31 (s, 3H),

3.65 (s, 3H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 9.9 (CH3), 14.7 (CH3), 22.3 (CH2), 25.2 (CH2), 27.0

3 Activated K2CO3: dried in the oven at 150 ºC overnight before use.

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(CH2), 32.2 (CH2), 34.0 (CH2), 34.7 (CH), 51.4 (CH3), 57.4 (CH3), 86.6 (CH), 174.2 (C) ppm. MS (CI)

for C12H24O3: m/z = 234.2 [M+NH4]+, HRMS (EI) calcd for C12H24O3-OCH3: 185.1541, found: 185.1540.

HO

O

(7R)-methoxy-(6S)-methylnonan-1-ol (18): To a solution of 9 (600 mg, 2.77 mmol) in dry THF (14 ml)

at -78 ºC, a solution of DIBAL-H (20% in toluene, 5.0 ml, 5.82 mmol, 2.1 eq) was added. The resulting

mixture was stirred overnight under nitrogen at -78 ºC, after which the reaction was quenched with an

aqueous solution of Rochelle salt and extracted with Et2O. The combined organic layers were washed

with brine (sat.), dried (MgSO4) and concentrated. The product was purified by column chromatography

(n-pentane-EtOAc 9:1) to give 18 (496 mg, 2.63 mmol, 95%) as a colorless oil. [α]D22 = – 3.5 º (c = 3.3,

CHCl3). 1H-NMR (CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.91 (t, J = 7.5 Hz, 3H), 1.10 (m, 1H),

1.20-1.76 (m, 10H, 1OH), 2.87 (m, 1H), 3.33 (s, 3H), 3.64 (m, 2H) ppm. 13C-NMR (CDCl3, 100.6 MHz)

δ = 9.9 (CH3), 14.7 (CH3), 22.3 (CH2), 26.0 (CH2), 27.2 (CH2), 32.5 (CH2), 32.7 (CH2), 34.7 (CH), 57.3

(CH3), 62.7 (CH2), 86.7 (CH) ppm. MS (CI) for C11H24O2: m/z = 206.2 [M+NH4]+, HRMS (EI) calcd for

C11H24O2-OCH3: 157.1592, found: 157.1589.

H

O O

(7R)-methoxy-(6S)-methylnonanal (4): To a solution of 18 (545 mg, 2.89 mmol) in dry DCM (15 ml),

Dess-Martin reagent (1.35 g, 3.18 mmol, 1.1 eq) was added and the resulting mixture was stirred under

nitrogen at room temperature for 30 min. The reaction was quenched with aq. Na2S2O3 (sat.) and aq.

NaHCO3 (sat.) and then extracted with DCM. The combined organic layers were washed with brine (sat.),

dried (MgSO4) and concentrated. The product was purified by column chromatography (n-pentane-

EtOAc 9:1) to give 4 (494 mg, 2.65 mmol, 92%) as a colorless oil. The product turned out to be unstable

and was therefore immediately used in the next reaction. [α]D22 = – 5.3 º (c = 3.1, CHCl3). 1H-NMR

(CDCl3, 400 MHz) δ = 0.79 (d, J = 6.8 Hz, 3H), 0.86 (t, J = 7.5 Hz, 3H), 1.06 (m, 1H), 1.22 (m, 1H),

1.30-1.47 (m, 4H), 1.50-1.66 (m, 3H), 2.39 (td, J = 1.8 Hz, J = 7.3 Hz, 2H), 2.82 (m, 1H), 3.28 (s, 3H),

9.72 (t, J = 1.8 Hz, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 9.8 (CH3), 14.7 (CH3), 22.3 (CH2), 27.0

(CH2), 32.2 (CH2), 34.7 (CH), 43.8 (CH2), 57.3 (CH3), 86.5 (CH), 202.6 (C) ppm.

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OOH

HONMe2

OH(+)-L*

(11R)-methoxy-(2,10S)-dimethyltridec-3-yne-(2,5R)-diol [(11R,10S,5R)-10]: (+)-L* (998 mg, 5.57

mmol, 2.1 eq) and Zn(OTf)2 (1.93 g, 5.31 mmol, 2.0 eq) were stirred under nitrogen for 15 min at room

temperature. Dry toluene (4.5 ml) and dry Et3N (0.78 ml, 5.57 mmol, 2.1 eq) were added and the resulting

mixture was stirred under nitrogen at room temperature for 2 h, after which 2-methyl-but-3-yn-2-ol (545

µl, 5.57 mmol, 2.1 eq) was added. After stirring for 15 min, a solution of 4 (494 mg, 2.65 mmol) in dry

toluene (5.7 ml) was added over 4 h by syringe pump and the resulting mixture was stirred overnight at

room temperature. The reaction was quenched with aq. NH4Cl (sat.), extracted with Et2O, washed with

brine (sat.) and dried (Na2SO4). The solvents were removed under reduced pressure and the product was

purified by column chromatography (n-pentane-EtOAc 7:3) to give (11R,10S,5R)-10 (563 mg, 2.08

mmol, 95% de, 78%) as a colorless oil. [α]D22 = – 5.1 º (c = 2.9, CHCl3). 1H-NMR (CDCl3, 400 MHz) δ =

0.84 (d, J = 6.8 Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H), 0.92 (m, 1H), 1.25 (m, 1H), 1.34-1.58 (m, 6H), 1.52 (s,

6H), 1.71 (m, 3H), 1.95 (s, 1OH), 2.87 (m, 1H), 3.34 (s, 3H), 4.39 (m, 1H) ppm. 13C-NMR (CDCl3, 100.6

MHz) δ = 10.0 (CH3), 14.8 (CH3), 22.3 (CH2), 25.5 (CH2), 27.1 (CH2), 31.3 (CH3), 32.5 (CH2), 34.8 (CH),

37.6 (CH2), 57.3 (CH3), 62.1 (CH), 64.9 (C), 83.1 (C), 86.7 (CH), 89.5 (C) ppm. MS (CI) for C16H30O3:

m/z = 288.3 [M+NH4]+, HRMS (EI) calcd for C16H30O3-CH3: 255.1960, found: 255.1961. The

diasteromeric ratio was measured by 19F-NMR of the corresponding Mosher ester (40:1, 95% de).

OOH

HONMe2

OH(-)-L*

(11R)-methoxy-(2,10S)-dimethyltridec-3-yne-(2,5S)-diol [(11R,10S,5S)-10]: (-)-L* (90 mg, 0.50

mmol, 2.1 eq) and Zn(OTf)2 (176 mg, 0.48 mmol, 2.0 eq) were stirred under nitrogen for 15 min at room

temperature. Then dry toluene (0.4 ml) and dry Et3N (70 µl, 0.50 mmol, 2.1 eq) were added and the

resulting mixture was stirred under nitrogen at room temperature for 2 h, after which 2-methyl-but-3-yn-

2-ol (49 µl, 0.50 mmol, 2.1 eq) was added. After stirring for 15 min, a solution of 4 (45 mg, 0.24 mmol)

in dry toluene (0.5 ml) was added over 4 h by syringe pump and the resulting mixture was stirred

overnight at room temperature. The reaction was quenched with aq. NH4Cl (sat.), extracted with Et2O,

washed with brine (sat.) and dried (Na2SO4). The solvents were removed under reduced pressure and the

product was purified by column chromatography (n-pentane-EtOAc 7:3) to give (11R,10S,5S)-10 (45 mg,

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0.17 mmol, 95% de, 70%) as a colorless oil. The 1H-NMR data were identical with those reported for

(11R,10S,5R)-10. The diasteromeric ratio was measured by 19F-NMR of the corresponding Mosher ester

(40:1, 95% de).

OO

HO

O

O

FF

F

(3,3,3)-trifluoro-(2R)-methoxy-(2R)-phenylpropionic acid 4-hydroxy-(1R)-[(6R)-methoxy-(5S)-

methyloctyl]-4-methylpent-2-ynyl [Mosher ester (R)-19]: To a solution of (11R,10S,5R)-10 (20 mg,

0.074 mmol) in dry pyridine (0.5 ml), DMAP (4.5 mg, 0.037 mmol, 0.5 eq) and (R)-(-)-MTPACl (18 µl,

0.096 mmol, 1.3 eq) were added and the resulting mixture was stirred under nitrogen at room temperature

for 2 h. The reaction was quenched with aq. NaHCO3 (sat.) and extracted with Et2O. The combined

organic layers were washed with brine (sat.), dried (MgSO4) and concentrated. The product was purified

by column chromatography (n-pentane-EtOAc 9:1) to give Mosher ester (R)-19 (33 mg, 0.067 mmol,

91%) as a colorless oil. 1H-NMR (CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H),

1.07 (m, 1H), 1.33-1.52 (m, 6H), 1.47 (s, 6H), 1.66 (m, 2H), 1.83 (m, 2H), 2.07 (s, 1OH), 2.85 (m, 1H),

3.32 (s, 3H), 3.55 (s, 3H), 5.52 (t, J = 6.8 Hz, 1H), 7.40 (m, 3H), 7.53 (m, 2H) ppm. 19F-NMR (CDCl3,

376 MHz) δ = – 72.17 (s, major diastereoisomer), – 71.88 (s, minor diastereoisomer) ppm (diasteromeric

ratio 40.0:1.0).

OO

HO

O

O

FF

F

(3,3,3)-trifluoro-(2R)-methoxy-(2R)-phenylpropionic acid 4-hydroxy-(1S)-[(6R)-methoxy-(5S)-

methyloctyl]-4-methylpent-2-ynyl [Mosher ester (S)-19]: To a solution of (11R,10S,5S)-10 (13 mg,

0.048 mmol) in dry pyridine (0.4 ml), DMAP (3 mg, 0.024 mmol, 0.5 eq) and (R)-(-)-MTPACl (12 µl,

0.062 mmol, 1.3 eq) were added and the resulting mixture was stirred under nitrogen at room temperature

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for 2 h. The reaction was quenched with aq. NaHCO3 (sat.) and extracted with Et2O. The combined


by column chromatography (n-pentane-EtOAc 9:1) to give Mosher ester (S)-19 (21 mg, 0.043 mmol,

90%) as a colorless oil. 1H-NMR (CDCl3, 400 MHz) δ = 0.81 (d, J = 6.8 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H),

1.03 (m, 1H), 1.32-1.53 (m, 6H), 1.50 (s, 6H), 1.63 (m, 2H), 1.79 (m, 2H), 2.01 (s, 1OH), 2.84 (m, 1H),

3.33 (s, 3H), 3.58 (s, 3H), 5.58 (t, J = 6.7 Hz, 1H), 7.40 (m, 3H), 7.56 (m, 2H) ppm. 19F-NMR (CDCl3,

376 MHz) δ = – 72.18 (s, minor diastereoisomer), – 71.88 (s, major diastereoisomer) ppm (diasteromeric

ratio 1:40).

OOTIPS

HO

(11R)-methoxy-(2,10S)-dimethyl-(5R)-triisopropylsilanyloxytridec-3-yn-2-ol (20): To a solution of 10

(563 mg, 2.08 mmol) in dry DCM (20 ml) at 0 ºC, 2,6-lutidine (0.36 ml, 3.12 mmol, 1.5 eq) and TIPSOTf

(0.73 ml, 2.70 mmol, 1.3 eq) were added and the resulting mixture was stirred under nitrogen at 0 ºC for

30 min. The reaction was quenched with aq. NaHCO3 (sat.) and then extracted with DCM. The combined


by column chromatography (n-pentane-EtOAc 95:5) to give 20 (843 mg, 1.98 mmol, 95%) as a colorless

oil. [α]D22 = + 17.1 º (c = 3.7, CHCl3). 1H-NMR (CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.91 (t, J

= 7.4 Hz, 3H), 0.98-1.17 (m, 21H), 1.24 (m, 2H), 1.33-1.54 (m, 6H), 1.49 (s, 6H), 1.68 (m, 3H), 1.89 (s,

1OH), 2.87 (m, 1H), 3.34 (s, 3H), 4.47 (t, J = 6.3 Hz, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.1

(CH3), 12.2 (CH), 14.7 (CH3), 18.0 (CH3), 22.3 (CH2), 25.3 (CH2), 27.2 (CH2), 31.3 (CH3), 32.5 (CH2),

34.7 (CH), 38.8 (CH2), 57.3 (CH3), 62.9 (CH), 65.0 (C), 83.9 (C), 86.7 (CH), 88.5 (C) ppm. MS (CI) for

C25H50O3Si: m/z = 444.2 [M+NH4]+, HRMS (EI) calcd for C25H50O3Si-C3H7: 383.2981, found: 383.2991.

OOTIPS

(9R)-methoxy-(8S)-methyl-(3R)-triisopropylsilanyloxyundec-1-yne (11): To a solution of 20 (843 mg,

1.98 mmol) in dry toluene (25 ml), NaH (60%, 95 mg, 2.38 mmol, 1.2 eq) was added and the resulting

mixture was refluxed under nitrogen overnight. The reaction was quenched with aq. NH4Cl (sat.) and then

extracted with EtOAc. The combined organic layers were washed with brine (sat.), dried (MgSO4) and

concentrated. The product was purified by column chromatography (n-pentane-Et2O 50:1) to give 11 (700

S10

mg, 1.90 mmol, 96%) as a colorless oil. [α]D22 = + 15.5 º (c = 2.8, CHCl3). 1H-NMR (CDCl3, 400 MHz) δ

= 0.83 (d, J = 6.8 Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H), 1.00-1.18 (m, 21H), 1.25 (m, 1H), 1.34-1.53 (m, 7H),

1.71 (m, 3H), 2.37 (d, J = 2.0 Hz, 1H), 2.87 (m, 1H), 3.33 (s, 3H), 4.47 (td, J = 2.0 Hz, J = 6.2 Hz, 1H)

ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.1 (CH3), 12.2 (CH), 14.7 (CH3), 18.0 (CH3), 22.3 (CH2),

25.2 (CH2), 27.3 (CH2), 32.6 (CH2), 34.7 (CH), 38.8 (CH2), 57.3 (CH3), 62.9 (CH), 72.0 (CH), 85.7 (C),

86.7 (CH) ppm. MS (CI) for C22H44O2Si: m/z = 386.3 [M+NH4]+, HRMS (EI) calcd for C22H44O2Si-

C3H7: 325.2563, found: 325.2557.

OOTIPS

22

(3R)-methoxy-(4S)-methyl-(9R)-triisopropylsilanyloxytetratriacont-10-yne (12): To a solution of 11

(355 mg, 0.96 mmol) in dry THF (6 ml) at -78 ºC, a solution of BuLi (1.6 M in hexane, 0.59 ml, 0.94

mmol, 0.98 eq)4 was added and the resulting mixture was stirred under nitrogen at -78 ºC for 10 min.

Then a solution of C23H47Br (500 mg, 1.25 mmol, 1.3 eq) in dry THF (3 ml) was added and the mixture

was allowed to warm up to room temperature. Activated NaI (36 mg, 0.24 mmol, 25 mol%)5 was added

and the resulting mixture was refluxed under nitrogen for 60 h. The reaction was quenched with aq.

NH4Cl (sat.) and then extracted with Et2O. The combined organic layers were washed with brine (sat.),

dried (MgSO4) and concentrated. The product was purified by column chromatography (n-pentane-Et2O

50:1) to give 12 (520 mg, 0.75 mmol, 87%)6 as a colorless oil. [α]D22 = + 10.3 º (c = 3.9, CHCl3). 1H-

NMR (CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.88 (t, J = 6.9 Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H),

0.96-1.52 (m, 70H), 1.67 (m, 4H), 2.17 (td, J = 1.8 Hz, J = 6.9 Hz 1H), 2.87 (m, 1H), 3.33 (s, 3H), 4.43 (tt,

J = 1.8 Hz, J = 6.2 Hz, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.2 (CH3), 12.3 (CH), 14.1 (CH3),

14.6 (CH3), 18.1 (CH3), 18.7 (CH2), 22.3 (CH2), 22.7 (CH2), 25.5 (CH2), 27.4 (CH2), 28.7 (CH2), 28.9

(CH2), 29.2 (CH2), 29.4 (CH2), 29.6 (CH2), 29.7 (CH2), 31.9 (CH2), 32.7 (CH2), 34.7 (CH), 39.3 (CH2),

57.3 (CH3), 63.2 (CH), 82.1 (C), 84.4 (C), 86.7 (CH) ppm.

4 When more than one eq. of BuLi was used, side reactions took place and the desired product was not formed, so it was

crucial to use 0.98 eq of base. 5 Activated NaI: recristalized in acetone and dried under reduced pressure at 100 ºC overnight before use. 6 Due to the amount of base used in the reaction (0.98 eq), 10% of the starting material was recovered after column

cromatography (35 mg). The reported yield takes in account this recovered starting material.

S11

OOH

22

(3R)-methoxy-(4S)-methyltetratriacont-10-yn-(9R)-ol (5): To a solution of 12 (500 mg, 0.72 mmol) in

dry THF (6 ml) at 0 ºC, a solution of TBAF (1.0 M in THF, 0.94 ml, 0.94 mmol, 1.3 eq) was added and

the resulting mixture was stirred under nitrogen at 0 ºC for 1 h. Then the THF was removed under

reduced pressure and the resulting crude was purified by column chromatography (n-pentane-EtOAc

95:5) to give 5 (354 mg, 0.66 mmol, 92%) as a colorless oil. [α]D22 = – 1.3 º (c = 4.1, CHCl3). 1H-NMR

(CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 6.9 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H), 0.98-

1.53 (m, 49H), 1.75-1.84 (m, 4H), 1.85 (d, J = 5.2 Hz, 1OH), 2.19 (td, J = 1.8 Hz, J = 7.0 Hz 1H), 2.86

(m, 1H), 3.33 (s, 3H), 4.34 (m, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.1 (CH3), 14.1 (CH3),


(CH2), 29.4 (CH2), 29.5 (CH2), 29.6 (CH2), 29.7 (CH2), 31.9 (CH2), 32.6 (CH2), 34.7 (CH), 38.2 (CH2),

57.3 (CH3), 62.7 (CH), 82.3 (C), 85.5 (C), 86.7 (CH) ppm. Positive ion ESI-HRMS calcd for C36H70O2Li

[M+Li]+: 541.5530, found: 541.5528.

OOHSiBn OOH

SiBn

2222

11-(benzyldimethylsilanyl)-(3R)-methoxy-(4S)-methyltetratriacont-10-en-(9R)-ol (14) and 10-

(benzyldimethylsilanyl)-(3R)-methoxy-(4S)-methyltetratriacont-10-en-(9R)-ol (15):

To a solution of 5 (225 mg, 0.42 mmol) in dry DCM (1.7 ml) at 0 ºC, benzyldimethylsilane (200 µl, 1.26

mmol, 3.0 eq) and [Cp*Ru(MeCN)3]PF6 (21.2 mg, 0.04 mmol, 10 mol%)7 were added and the resulting

mixture was stirred under nitrogen at room temperature for 2 h. Then the DCM was removed under

reduced pressure and the resulting crude was purified by column chromatography (n-pentane-EtOAc

95:5) to give a mixture of regioisomers 14:15 4.0:1.0 (248 mg, 0.36 mmol, 86%) as a colorless oil. A

pure fraction of 14 was isolated and characterized: [α]D22 = + 7.9 º (c = 4.6, CHCl3). 1H-NMR (CDCl3,

400 MHz) δ = 0.15 (s, 3H), 0.16 (s, 3H), 0.83 (d, J = 6.8 Hz, 3H), 0.89 (t, J = 6.9 Hz, 3H), 0.91 (t, J = 7.4

Hz, 3H), 1.02-1.57 (m, 52H), 1.69 (m, 1H), 2.05 (m, 2H), 2.21 (s, 2H), 2.87 (m, 1H), 3.33 (s, 3H), 4.04

(m, 1H), 5.91 (d, J = 9.4 Hz, 1H), 7.01 (d, J = 7.8 Hz, 2H), 7.09 (t, J = 7.2 Hz, 1H), 7.22 (t, J = 7.2 Hz,

2H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = -1.4 (CH3), -1.0 (CH3), 10.1 (CH3), 14.1 (CH3), 14.7 7 [Cp*Ru(MeCN)3]PF6 was weight in a glove box.

S12

(CH3), 22.3 (CH2), 22.7 (CH2), 25.9 (CH2), 26.9 (CH2), 27.6 (CH2), 29.4 (CH2), 29.5 (CH2), 29.7 (CH2),

30.5 (CH2), 31.9 (CH2), 32.6 (CH2), 34.7 (CH), 37.3 (CH2), 38.1 (CH2), 57.3 (CH3), 71.0 (CH), 86.7 (CH),

124.3 (CH), 128.1 (CH), 128.3 (CH), 139.8 (C), 140.9 (C), 145.8 (CH) ppm. Positive ion ESI-HRMS

calcd for C45H85O2Si [M+H]+: 685.6313, found: 685.6316.

OOHO

22

OOH

22O

(9R)-hydroxy-(3R)-methoxy-(4S)-methyltetratriacontan-11-one (16) and (9R)-hydroxy-(3R)-

methoxy-(4S)-methyltetratriacontan-10-one (21): To a solution of 14:15 4.0:1.0 (248 mg, 0.36 mmol)

in dry THF (2.5 ml) at 0 ºC, a solution of TBAF (1.0 M in THF, 0.43 ml, 0.43 mmol, 1.2 eq) was added

and the resulting mixture was stirred under nitrogen at 0 ºC for 20 min. Then methanol (2.5 ml) was

added, followed by KHCO3 (108 mg, 1.08 mmol, 3.0 eq) and H2O2 (30%, 0.6 ml, 5.43 mmol, 15.0 eq)

and the resulting mixture was stirred at room temperature overnight. Water was added to the reaction

mixture and then extracted with EtOAc. The combined organic layers were washed with brine (sat.), dried

(MgSO4) and concentrated. The crude was purified by column chromatography (n-pentane-EtOAc 95:5)

to give 16 (126 mg, 0.23 mmol, 63%) as a white solid and 21 (30 mg, 0.05 mmol, 15%) as a white solid.

16: [α]D22 = – 17.4 º (c = 0.9, CHCl3), mp = 186-188 ºC. 1H-NMR (CDCl3, 400 MHz) δ = 0.82 (d, J = 6.8

Hz, 3H), 0.87 (t, J = 6.9 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H), 0.99-1.75 (m, 53H), 2.40 (t, J = 7.5 Hz, 2H),

2.47 (dd, J = 9.2 Hz, J = 17.5 Hz, 1H), 2.58 (dd, J = 2.7 Hz, J = 17.5 Hz, 1H), 2.85 (m, 1H), 3.09 (d, J =

2.7 Hz, 1OH), 3.32 (s, 3H), 4.01 (m, 1H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.0 (CH3), 14.1


29.4 (CH2), 29.6 (CH2), 29.7 (CH2), 31.9 (CH2), 32.6 (CH2), 34.7 (CH), 36.4 (CH2), 43.7 (CH2), 48.9

(CH2), 57.4 (CH3), 67.6 (CH), 86.6 (CH), 212.6 (C) ppm. Positive ion ESI-HRMS calcd for C36H72O3Li

[M+Li]+: 559.5636, found: 559.5634.

21: 1H-NMR (CDCl3, 400 MHz) δ = 0.83 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 6.9 Hz, 3H), 0.90 (t, J = 7.4 Hz,

3H), 1.04-1.72 (m, 53H), 1.83 (m, 1H), 2.02 (m, 1H), 2.44 (m, 2H), 2.86 (m, 1H), 3.33 (s, 3H), 3.50 (d, J

= 4.9 Hz, 1OH), 4.16 (m, 1H) ppm.

S13

OOHOH

22

OOHOH

22

(3R)-methoxy-(4S)-methyltetratriacontane-(9R,11R)-diol (anti-3, phthiocerol) and (3R)-methoxy-

(4S)-methyltetratriacontane-(9R,11S)-diol (syn-3): A solution of Me4N(AcO)3BH (190 mg, 0.72 mmol,

5.0 eq) in dry THF (1.0 ml) and dry AcOH (1.0 ml) was stirred under nitrogen at room temperature for 30

min. Then a solution of 16 (80 mg, 0.14 mmol) in dry THF (1.0 ml) was added and the resulting mixture

was stirred under nitrogen at room temperature overnight. The reaction was quenched with an aqueous

solution of Rochelle salt and extracted with Et2O. The combined organic layers were washed with brine

(sat.), dried (MgSO4) and concentrated. The product was purified by column chromatography (n-pentane-

EtOAc 8:2) to give 3 as a mixture of anti:syn 88:12 (70 mg, 0.13 mmol, 90%) as a white solid. Both diols

were separated by a second column chromatography (n-pentane-EtOAc 9:1) and phthiocerol was isolated

(58 mg, 0.11 mmol, 77%) as a white solid.

syn-3: mp = 65-67 ºC. 1H-NMR (CDCl3, 400 MHz) δ = 0.82 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 6.9 Hz, 3H),

0.91 (t, J = 7.4 Hz, 3H), 1.05-1.73 (m, 57H), 2.87 (m, 1H, 1OH), 2.95 (s, 1OH), 3.33 (s, 3H), 3.84 (m,

2H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ = 10.0 (CH3), 14.1 (CH3), 14.8 (CH3), 22.3 (CH2), 22.7


34.8 (CH), 38.3 (CH2), 42.9 (CH2), 57.4 (CH3), 73.2 (CH), 73.3 (CH), 86.7 (CH) ppm. Positive ion ESI-

HRMS calcd for C36H74O3Li [M+Li]+: 561.5793, found: 561.5789.

anti-3 (phthiocerol): [α]D22 = – 4.5 º (c = 0.4, CHCl3), mp = 72-74 ºC. 1H-NMR (CDCl3, 400 MHz) δ =

0.82 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 6.9 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H), 0.99-1.75 (m, 57H), 2.61 (s,

1OH), 2.66 (s, 1OH), 2.86 (m, 1H), 3.32 (s, 3H), 3.91 (m, 2H) ppm. 13C-NMR (CDCl3, 100.6 MHz) δ =


(CH2), 29.6 (CH2), 29.7 (CH2), 31.9 (CH2), 32.6 (CH2), 34.7 (CH), 37.4 (CH2), 37.5 (CH2), 42.2 (CH2),

57.3 (CH3), 69.3 (CH), 69.4 (CH), 86.7 (CH) ppm. Positive ion ESI-HRMS calcd for C36H74O3Li

[M+Li]+: 561.5793, found: 561.5790.

S14

OOO

OO

19 19

22

PDIM A (1): anti-3 (4.0 mg, 7.21 µmol), mycocerosic acid (10.4 mg, 21.62 µmol, 3.0 eq), DCC (6.0 mg,

28.84 µmol, 4.0 eq) and DMAP (3.5 mg, 28.84 µmol, 4.0 eq) were dissolved in dry DCM (0.4 ml) and the

resulting mixture was stirred under nitrogen at room temperature overnight. The solvent was removed

under reduced pressure and the product was purified by column chromatography (n-pentane-Et2O 50:1) to

give PDIM A as a white wax (6.7 mg, 4.54 µmol, 63%). 1H-NMR (CDCl3, 500 MHz) δ = 0.74-1.81 (m,

190H), 2.54 (m, 2H), 2.87 (m, 1H), 3.34 (s, 3H), 4.85 (m, 2H) ppm. 13C-NMR (CDCl3, 125 MHz) δ =

10.1, 14.1, 14.7, 18.4, 20.4, 20.5, 20.7, 22.3, 22.7, 25.2, 25.6, 27.0, 27.2, 27.5, 28.0, 29.4, 29.5, 29.6, 29.7,

29.8, 29.9, 30.1, 31.9, 32.6, 34.8, 36.6, 37.8, 38.4, 41.0, 45.3, 45.5, 57.4, 70.3, 86.6, 175.9, 176.0 ppm.

MALDI-TOF for C100H198O5: m/z = 1503.6 [M+Na]+, 1519.3 [M+K]+.

S15

O

17

ppm (f1)1.02.03.04.05.06.07.0

3.00

4.04

1.091.01

2.02

3.14

O

6

ppm (f1)1.02.03.04.05.06.07.0

3.00

3.00

1.00

1.01

0.94

0.90

5.99

2.03

S16

O

6

ppm (f1)50100150200

ppm (f1)50100150200

S17

OO

7

ppm (f1)1.02.03.04.05.06.07.0

1.00

8.06

5.97

2.00

1.19

ppm (f1)050100150

S18

MeO

O OH

8

ppm (f1)1.02.03.04.05.06.07.0

1.00

2.03

2.98

5.89

10.28

ppm (f1)50100150

ppm (f1)50100150

S19

MeO

O O

9

ppm (f1)1.02.03.04.05.06.07.0

3.03

1.00

3.02

2.04

3.08

3.07

4.09

1.051.08

2.99

ppm (f1)50100150

ppm (f1)50100150

S20

HO

O

18

ppm (f1)1.02.03.04.05.06.07.0

2.05

3.05

1.00

5.81

11.06

1.08

ppm (f1)102030405060708090

ppm (f1)102030405060708090

S21

H

O O

4

ppm (f1)0.05.010.0

2.98

1.00

1.98

3.134.01

1.071.06

6.08

1.00

ppm (f1)50100150200

ppm (f1)50100150200

S22

OOH

HO

10

ppm (f1)1.02.03.04.05.06.07.0

1.00

1.09

2.98

1.04

6.08

12.00

1.10

1.01

3.27

ppm (f1)102030405060708090

ppm (f1)50

S23

OO

HO

O

O

FF

F

(R)-19

ppm (f1)1.02.03.04.05.06.07.0

2.033.02

3.06

3.03

1.10

1.04

1.98

1.95

12.08

0.96

1.00

6.10

S24

OO

HO

O

O

FF

F

(R)-19

ppm (f1)-80.0-75.0-70.0-65.0-60.0

1.0040.65

S25

OO

HO

O

O

FF

F

(S)-19

ppm (f1)1.02.03.04.05.06.07.0

1.00

2.04

3.00

3.08

2.97

1.03

0.96

2.032.05

12.47

1.10

6.55

S26

OO

HO

O

O

FF

F

(S)-19

ppm (f1)-80.0-75.0-70.0-65.0-60.0

39.971.00

S27

OOTIPS

HO

20

ppm (f1)1.02.03.04.05.06.07.0

1.06

3.02

21.23

1.02

1.00

2.98

6.14

2.01

12.06

ppm (f1)50

S28

OOTIPS

HO

20

ppm (f1)102030405060708090

S29

OOTIPS

11

ppm (f1)1.02.03.04.05.06.07.0

1.00

3.08

1.03

0.99

7.04

21.31

3.003.00

1.18

2.98

ppm (f1)50

S30

ppm (f1)50

OOTIPS

1222

ppm (f1)1.02.03.04.05.06.07.0

3.01

1.00

1.00

2.08

9.05

4.04

70.02

S31

ppm (f1)1020304050607080

ppm (f1)1020304050607080

S32

OOH

522

ppm (f1)1.02.03.04.05.06.07.0

1.00

3.01

2.09

0.98

9.12

49.09

3.83

1.05

ppm (f1)1020304050607080

S33

O

14

OHSiBn

22

ppm (f1)0.01.02.03.04.05.06.07.0

1.03

2.01

0.99

1.06

3.00

1.00

5.98

51.66

1.12

2.05

2.08

2.05

9.06

ppm (f1)050100

S34

O

16

OHO

22

ppm (f1)1.02.03.04.05.06.07.0

3.00

1.03

1.02

1.04

2.031.14

52.71

9.02

1.03

ppm (f1)50100150200

ppm (f1)50100150200

S35

OOH

21

22O

ppm (f1)1.02.03.04.05.06.07.0

1.05

3.16

1.00

53.22

9.04

1.04

1.98

1.07

1.04

S36

OOHOH

syn-3

22

ppm (f1)1.02.03.04.05.06.07.0

3.19

2.00

9.00

1.012.01

57.33

ppm (f1)50

S37

OOHOH

anti-3

22

ppm (f1)1.02.03.04.05.06.07.0

2.00

56.81

9.05

1.06

2.01

3.04

ppm (f1)102030405060708090

S38

OOO

OO

PDIM A (1)

19 19

22

ppm (f1)1.02.03.04.05.06.07.0

2.18

190.02

2.94

0.99

2.06

ppm (f1)50100150

S39

OOO

OO

PDIM A (1)

19 19

22

MALDI-TOF mass spectra of PDIM A (1)

1 µl of a 1 mg/ml solution was mixed with 0.5 µl of the matrix solution.

The matrix used was 2,5-dihydroxybenzoic acid (10 mg/ml) in CHCl3/MeOH (1:1 v/v).

[M+Na]+

[M+K]+

supporting information · 2zn (1.2 m in toluene, 2.3 ml, 2.72 mmol, 1.5 eq) was added dropwise over...

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