1
[Supplementary Information A]
Supplementary figure 1 A representative chromatogram of columns, molecular
structure and fragmentation patterns of daumone.
a, Daumone activity eluted from the amine column. The chromatogram was detected on
optical density 380nm (OD380).
Each fraction applied to ESI-MSD, and then %
daumone determinate by dauer assay. The open circle represents a faction that was
collected and quantified for dauer-forming activity. The sample solution is loaded onto
the amine column, which is equilibrated with isopropanol solution. The daumone
fraction bound to the column was eluted with a gradient solution from isopropanol to
distilled water. b, Daumone activity eluted from the GPC column. The chromatogram
was detected on optical density 254 nm (OD254) and reflective index detector (RI).
Each fraction applied to ESI-MSD, and then % daumone determinate by dauer assay.
The open circle represents a faction that was collected and quantified for dauer-forming
activity.
The sample solution is loaded onto the GPC (W251) column, which is
equilibrated with methanol solution. c, Positive ion mass spectrum of the fraction
diluted in methanol. d, Positive ion mass spectrum of the fraction diluted in 0.1mM
ammonium acetate in methanol. e, Negative ion mass spectrum of the fraction diluted
in methanol. These spectra were obtained at low resolution. f, High resolution MS/MS
spectrum obtained by collisionally activated dissociation of sodiated daumone with m/z
299 using a Q-TOF mass spectrometer. Argon was used as the collision gas, and
collision energy of 30 eV applied.
Supplementary figure 2 DEPT spectrum of the daumone.
2
The
13
C NMR showed the daumone carbons, which were analyzed from the DEPT
spectrum to consist of two methyls (CH3), five methylenes (CH2) and five methines
(CH).
Supplementary figure 3 2D 1H, 13C HMBC spectrum of the daumone.
The corresponding parts of the 1H and
13
C NMR spectra are displayed along the
horizontal and vertical axes, respectively. The linkage positions were determined by
identifying the cross peaks from the HMBC spectrum, which gives cross peaks with
covalently bound H and C. Carbon chemicalshifts were assigned from the HMQC
spectrum based on proton resonances. HMBC experiment to emphasize the long range
coupling, either 2J(C,H) or 3J(C,H) between the carbons and protons.
In the HMBC
spectrum, the methylene protons gave cross-peaks with the carboxyl carbon at  = 177.3.
As two groups of protons in –C5H2-C6H- and –C2’H-C3’H- are not correlation with
each other, some coupling network is not observed.
Supplementary figure 4 2D TOCSY spectrum of the daumone.
The mixing time of 53 ms dissolved in 99.9% d-MeOH. Though bond connectivity
observed in the TOCSY spectrum. As two groups of protons in –C5H2-C6H- and –
C2’H-C3’H- are not bond connectivity with each other.
Supplementary figure 5 The observed ROEs of the daumone.
The sequence of atoms is arranged by their relative order. Distance was denoted by a
gray scale. Distances are obtained by calibration of the cross-peak intensities between
germinal protons (4’). Proton distances calculated using the equation: rij = rref (Iref/Iij)-6
3
(1), where rij is the distance and Iij is the peak intensity between atom i and atom j. (1)
Croasmun, W.R. and Carlson, R.M.K. (1994) Two-Dimensional NMR Spectroscopy:
Applications for Chemists and Biochemists. 2nd Ed., VCH Publishers, Inc.
Supplementary figure 6 500MHz 1H NMR spectrum of compound 1 (synthetic) at
298 K.
Supplementary figure 7 500MHz 1H NMR spectrum of compound 1 (natural) at
298 K.
Supplementary figure 8 125.7MHz 13C NMR spectrum of compound 1 at 298 K.
Supplementary figure 9 500MHz DEPT spectrum of compound 1 at 298 K.
Supplementary figure 10 500MHz HMBC spectrum of compound 1 at 298 K.
Supplementary figure 11 500MHz ROESY spectrum of compound 1 at 298 K
Supplementary figure 12 500MHz NOESY spectrum of compound 1 at 298 K.
Supplementary figure 13 FT-IR spectrum of compound 1 at 298 K.
Supplementary figure 14 HRMS(FAB) spectrum of compound 1 at 298 K.
4
Supplementary figure 15 High resolution Q-TOF mass spectrum of compound 1.
5
[Supplementary Information B]
a. Summary of daumone chemical synthesis
Stereospecific total synthesis of the pheromone was designed and successively achieved
from L-rhamnose in 10 steps (Fig. 2). Key intermediate (8) was synthesized from Lrhamnose (2) as depicted (Fig. 2)1,2,3. Thus, protection of all four hydroxyl groups of Lrhamnose (2) with benzoyl chloride/pyridine and subsequent selective deprotection of
benzoyl group at C-2’of compound (3) with ammonia afforded compound (4) with the
natural α-configuration at C-2’exclusively (14 : 1) in 87% yield. Without separation of
diastereomers at C-2’, oxidation of anomeric hemiacetal at C-2’ of compound (4) with
PCC in the presence of molecular sieves afforded the ketone (5) in 85 % yield. Selective
elimination of benzoyl group at C-4’ of compound (5) with triethylamine afforded
α , β-unsaturated double
compound (6
bond of compound (6) in the presence of 10% Pd/C gave compound (7) with natural configuration of the O-benzyl group at C-3’(yield 85%).
The stereoselectivity of
hydrogenation may be ascribed to the steric hindrance exerted by the quasi axial C-7
methyl group in the conformation of (6) that would prevent the attack of hydrogen from
underneath the ring. Reduction of (7) with disiamylborane in THF afforded the key
intermediate (8) in 93.8 % yield in the ratio of 2’R/2’S = 4.6/1. Preparation of the
second key intermediate (10) is described in Fig. 2.4,5 Regiospecific Grignard-type
alkylation of readily available (R)-(+)-propylene oxide (9) with 5-bromo-1-pentene and
magnesium cleanly afforded 7(R)-hydroxy-1-octene (10)5 [α]D20 = - 11.11 (c 4.30,
acetone), lit.5 (S-form) [α]D20 = + 11.20 (c 4.30, acetone), in 65% yield without
racemization.
The coupling of the key intermediate (8) with compound (10) is
described in Fig. 2. Thus, acetalization of (8) with compound (10) in the presence of
6
BF3.Et2O provided compound (11) in 72% yield with the natural α-configuration at C-2
stereoelectively. Final oxidation and deprotection of compound (11) to the target
compound (1) are shown in Fig. 2. Thus, oxidation of the terminal double bond of
compound (11) into compound (12) was achieved with KMnO4 in the presence of
NaHCO3 in one step (yield 87 %).6,7 Debenzoylation of compound (12) with sodium
methoxide and subsequent in situ acidification in the presence of acidic resin (Amberlite
IR-10, H+) successively furnished the target compound, (-)-6R-(3’R,5’R-dihydroxy-6’Smethyltetrahydro-pyran-2’R-yloxy) heptanoic acid (1)8 in 87% yield. The synthetic
compound (1) was identical with natural daumone isolated from C. elegans with respect
to spectroscopic data: 2D-NMR(500 MHz), 13C NMR, IR, HRMass and specific
rotation [α]D25 = - 81.0 (c 0.1, CH3OH), respectively. Total synthesis of compound (1)
finally confirmed the absolute stereochemistry of natural daumone as 6R, 2’R, 3’R, 5’R,
6’S, respectively. The amount of synthetic daumone for effective dauer formation was
indentical to natural daumone (i.e., 384 μM of daumone in the presence of 160 μg/plate
of dead E. coli).
(1) Han, O.; Liu, H. W. Synthesis of Stereospecifically labeled Carbohydrates:
Preparation of (3S)- and (3R)-[3-2H1]ascarylose, Tetrahedron Lett. 28, 1073-1076
(1987)
(2) Russell, R. N.; Weigel, T. M.; Han, O. S.; Liu, H. W. Synthesis of Stereospecifically
Labelled 3,6-Dideoxyhexoses, Carbohydr. Res., 201, 95-114 (1990)
(3) Varela, O. J.; Cirelli, A. F.; de Lederkremer, R. M. β-Elimination in Aldonolactones.
Synthesis of 3,6-Dideoxy-L-arabino-Hexose, Carbohydr. Res., 70, 27-35 (1979)
7
(4) Lee, W. W.; Shin, H. J.; Chang, S. B. A rapid formal synthesis of the macrolide (−)A26771B, Tetrahedron : Asymmetry, 12, 29-31 (2001)
(5) Fürstner, A.; Seidel, G.; Kindler, N. Macrocycles by Ring-Closing-Metathesis, XI:
Syntheses of (R)-(+)-Lasiodiplodin, Zeranol and Truncated Salicylihalamides,
Tetrahedron, 55, 8215-8230 (1999)
(6) Jung, M.; Lee, K.; Kendrick, H.; Robinson, B. L.; Croft, S. L. Synthesis, Stability
and Antimalarial Activity of New Hydrolytically Stable and Water Soluble
(+)Deoxoartelinic Acid, J. Med. Chem., 45, 4940 –4944 (2002)
(7) Jung, M.; Lee, S.; Ham, J.; Lee, K.; Kim, H.; Kim, S. K. Antitumor Activity of
Novel Deoxoartemisinin Monomer, Dimers and Trimer, J. Med. Chem., 46, 987-994
(2003)
(8) 1 ; a colorless syrup, Rf = 0.43 (EtOAc / MeOH, 11:1, v/v); [α]D25 = - 81.0 (c 0.1,
MeOH); FT-IR (film) vmax 3391, 2969, 2933, 1712, 1452, 1379, 1244, 1126, 1103, 1042,
1031 cm-1; 1H NMR (500 MHz, CD3OD) δ 4.64 (s, 1H, H-2), 3.80-3.77 (m, 1H, H-6),
3.72-3.71 (m, 1H, H-3), 3.63-3.59 (m, 1H, H-6), 3.54-3.49 (m, 1H, H-5), 2.30 (t, 2H, J
= 7.5 Hz, H-2), 1.96-1.92 (m, 1H, H-4eq), 1.79-1.74 (m, 1H, H-4ax), 1.61 (m, 2H, H3), 1.56-1.50 (m, 2H, H-5), 1.47 (m, 2H, H-4), 1.21 (d, 3H, J = 6.5 Hz, H-7), 1.12 (d,
3H, J = 6.5 Hz, H-7); 13C NMR (125.7 MHz, CD3OD) δ 177.7(C-1), 97.6(C-2, α),
72.4(C-6), 71.3(C-6), 70.1(C-3), 68.5(C-5), 38.2(C-5), 36.1(C-4), 35.0(C-2), 26.5(C3), 26.1(C-4), 19.4(C-7), 18.2(C-7); HRMS(FAB) calcd for C13H25O6 (M+ + H) m/z
277.1651, found 277.1652.
8
b. Supporting materials for the chemical synthesis of daumone
General methods.
All other commercial reagents and solvents were used as received without
further purification.
Reaction solvents were distilled from calcium hydride for
dichloromethane and from sodium metal and benzophenone for tetrahydrofuran. The
reactions were monitored and the Rf values determined using analytical thin layer
chromatography (TLC) with Merck silica gel 60, F-254 precoated plates (0.25 mm
thickness). Spots on the TLC plates were visualized using ultraviolet light (254 nm), a
basic potassium permanganate solution or cerium sulfate/ Ammonium dimolybdate/
H2SO4 solution followed by heating on a hot plate. Flash column chromatography was
performed with Merck silica gel 60 (230-400 mesh).
1
H NMR spectra were recorded on Bruker DPX-250 or Varian Unity-Inova 500
Spectrometers. Proton chemical shifts are reported in ppm (δ) relative to internal
tetramethylsilane (TMS, δ 0.0) or with the solvent reference relative to TMS employed
as the internal standard (CDCl3, δ 7.26 ppm; d4-CD3OD, δ 3.31 ppm). Data are reported
as follows: chemical shift (multiplicity [singlet (s), doublet (d), triplet (t), quartet (q),
and multiplet (m)], coupling constants [Hz], integration).
13
C NMR spectra were
recorded on Bruker DPX-250 (62.9 MHz) or Varian Unity-Inova 500 (125.8 MHz)
spectrometers with complete proton decoupling. Carbon chemical shifts are reported in
ppm (δ) relative to TMS with the respective solvent resonance as the internal standard
(CDCl3, δ 77.0 ppm; d4-CD3OD, δ 49.0 ppm). Infrared (IR) spectra were recorded on a
Nicolet Model Impact FT-IR 400 spectrometer. Data are reported in wave numbers
(cm-1). Specific rotation was recorded on a RUDOLPH Research AUTOPOL® III
9
automatic polarimeter. High Resolution Mass spectrometer was recorded on a JEOL
JMS-AX505WA, HP 5890 Series II. Elemental Analysis (EA) was recorded on a Carlo
Erba Instruments EA1L08-Elemental Analyzer.
OBz
O
BzO
BzO
OBz
Compd. 3
2,3,4,5-Tetra-O-benzoyl-L-rhamnopyranose (3)
To a solution of L-rhamnose monohydrate 2 (7.5g, 41.2mmol) in dry pyridine (100mL)
at 0°C, benzoyl chloride (28.7 mL, 0.247 mol) was added slowly. The solution was
warmed to r. t. and stirred for 16 h. and quenched by addition of H2O (15mL). The
reaction mixture was extracted by CH2Cl2 (50mL×2), followed washed by 1M HCl
(40mL×2) and successively by saturated NaHCO3 (40mL) and dried by anhydrous
MgSO4. Concentration in vacuo and purification by flash column chromatography
(toluene / EtOAc, 10:1, v/v) afforded compound 3 (22.7g, 95%, α:β = 2:1).
3α; an amorphous white solid, Rf = 0.58 (Toluene / EtOAc, 10:1, v/v); [α]D22= +82.0 (c
1.5, CHCl3) ; FT-IR (film) vmax 3066, 3032, 2986, 1730, 1601, 1452, 1260, 1176, 1094,
1068, 1027, 965 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.22-7.25 (m, 20H, aromatic H),
6.57 (d, 1H, J = 1.6 Hz, H-2), 6.01 (dd, 1H, J = 3.4, 10.2 Hz, H-4), 5.89 (dd, 1H, J =
1.9, 3.2 Hz, H-3), 5.82 (t, 1H, J = 10.0 Hz, H-5), 4.41-4.35 (m, 1H, H-6), 1.42 (d, 3H,
10
J = 6.2 Hz, H-7); 13C NMR (62.9 MHz, CDCl3) δ 165.8(2), 165.4, 164.1, 134.0, 133.8,
133.6, 133.4, 130.2(2), 130.1(2), 129.8(4), 129.1(2), 129.0(2), 128.8(2), 128.7(2),
128.6(2), 128.4(2), 91.4(C-2, α), 71.3, 70.0, 69.8, 69.4, 17.8(C-7).; HRMS(FAB) calcd
for C34H28NaO9 (M+ + Na) m/z 603.1631, found 603.1637.; Anal. Calcd for C34H28O9: C,
70.34; H, 4.86. Found: C, 70.31; H, 4.83.
3β; an amorphous white solid, Rf = 0.53 (Toluene/EtOAc, 10:1, v/v); [α]D22= +139.0 (c
1.0, CHCl3); FT-IR (film) vmax 3066, 3031, 2985, 2937, 1731, 1601, 1452, 1265, 1178,
1106, 1069, 1027 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.21-7.22 (m, 20H, aromatic H),
6.39 (s, 1H), 6.11 (s, 1H), 5.77 (s, 1H), 5.74 (s, 1H), 4.14-4.08 (m, 1H, H-6), 1.49 (d,
3H, J = 6.1 Hz, H-7); 13C NMR (62.9 MHz, CDCl3) δ 165.8, 165.7, 165.5, 164.3, 133.8,
133.6(2), 133.4, 130.2(2), 130.1(2), 130.0(2), 129.8(2), 129.5, 129.1, 128.9, 128.7(3),
128.6(2), 128.5(2), 128.4(2), 91.3(C-2, β), 71.9, 71.5, 71.3, 69.7, 17.8(C-7).
OH
O
BzO
BzO
OBz
Compd. 4
3,4,5-Tri-O-benzoyl-L-rhamnopyranose (4)
To a cooled solution of compound 3 (22.4g, 38.6mmol) in MeOH and THF (3:7,
400mL) at 0°C, NH3 gas was dissolved by bubbling for 15 min. and stirred for 1 h. at r.
t. The reaction was monitored by TLC and completed after 12 h. Concentration in vacuo
11
and purification by flash column chromatography (toluene / EtOAc, 10:1, v/v) afforded
compound 4 (16g, 87%, α:β = 14:1).
4α; a white solid, Rf = 0.18 (Toluene / EtOAc, 10:1, v/v); [α]D23= +236.0 (c 1.0, CHCl3);
FT-IR (film) vmax 3458, 3062, 2985, 2935, 1727, 1601, 1451, 1348, 1264, 1102, 1069,
1027 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.12-7.22 (m, 15H, aromatic H), 5.95 (dd, 1H,
J = 3.2, 10.1 Hz, H-4), 5.74-5.62 (m, 2H), 5.49-5.48 (m, 1H), 4.54-4.43 (m, 1H, H-6),
4.21 (d, 1H, J = 4.0 Hz, -OH), 1.37 (d, 3H, J = 6.2 Hz, H-7);
13
C NMR (62.9 MHz,
CDCl3) δ 166.0, 165.9, 165.8, 133.6, 133.5, 133.3, 130.0(2), 129.9(2), 129.8(2), 129.4,
129.3, 129.2, 128.7(2), 128.5(2), 128.4(2), 92.3(C-2, α), 72.1, 71.5, 69.9, 66.7, 17.8(C7).; HRMS(FAB) calcd for C27H24NaO8 (M+ + Na) m/z 499.1369, found 499.1372.;
Anal. Calcd for C27H24O8: C, 68.06; H, 5.08. Found: C, 68.05; H, 4.98.
O
O
BzO
BzO
OBz
Compd. 5
3,4,5-Tri-O-benzoyl-L-rhamnono-2,6-lactone (5)
Dry CH2Cl2 (250mL) was added to the preloaded PCC (30g, 0.139mmol) and dried 4Å
molecular sieves (25g) under N2, and the solution was stirred for 1 h. and cooled to 0°C.
Compound 4 (16g, 33.6mmol) in dry CH2Cl2 (250mL) was added to the solution and
warmed to r. t. and stirred for 4 h. The reaction was completed by addition of Et2O
(200mL) and the reaction solution was filtered through silica gel. Concentration in
12
vacuo and purification by flash column chromatography (Toluene / EtOAc, 10:1, v/v)
afforded compound 5 (13.54g, 85%).
5; an amorphous white solid, Rf = 0.51 (Toluene/EtOAc, 10:1, v/v); [α]D22= -10.0 (c 0.5,
CHCl3); FT-IR (film) vmax 3064, 3031, 2983, 2936, 1784, 1730, 1601, 1452, 1393, 1259,
1096, 1026 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.10-7.29 (m, 15H, aromatic H), 6.28
(d, 1H, J = 3.8 Hz), 6.05 (dd, 1H, J = 1.4, 3.8 Hz), 5.34 (dd, 1H, J = 1.4, 11.0 Hz), 4.964.85 (m, 1H, H-6), 1.61 (d, 3H, J = 6.3 Hz, H-7);
C NMR (62.9 MHz, CDCl3) δ
13
165.9(C-2), 165.1, 164.9, 164.8, 134.0, 133.9, 133.8, 130.1(4), 130.0(2), 128.7(5),
128.5(3), 128.4, 74.8, 74.1, 71.8, 67.6, 19.0(C-7).; HRMS (FAB) calcd for C27H23O8
(M+ + H) m/z 475.1393, found 475.1393.; Anal. Calcd for C27H22O8: C, 68.35; H, 4.67.
Found: C, 68.39; H, 4.76.
O
BzO
O
OBz
Compd. 6
3,5-Di-O-benzoyl-4,7-dideoxy-L-erythro-hex-3-enono-2,6-lactone(6)
A solution of 5 (13.2g, 27.8mmol) in Et3N : CHCl3 (1:4, 500mL) was stirred under N2
for 16h. The reaction mixture was extracted by H2O, and dried by anhydrous MgSO4.
Concentration in vacuo and purification by flash column chromatography (Toluene /
EtOAc, 10:1, v/v) afforded compound 6 (6.37g, 65%).
13
6; a crystalline white solid, Rf = 0.53 (Toluene/EtOAc, 10:1, v/v); mp 108-112°C ;
[α]D21= -93.1 (c 1.0, CHCl3); FT-IR (film) vmax 3069, 3007, 2936, 2920, 1738, 1674,
1598, 1452, 1355, 1257, 1155, 1115, 1060 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.137.44 (m, 10H, aromatic H), 6.71 (d, 1H, J = 4.3 Hz, H-4), 5.69 (t, 1H, J = 4.7 Hz, H-5),
5.00-4.90 (m, 1H, H-6), 1.64 (d, 3H, J = 6.7 Hz, H-7); 13C NMR (62.9 MHz, CDCl3) δ
165.5, 164.3, 158.0(C-3), 140.8, 134.3, 133.9, 130.5(2), 130.0(2), 128.7(5), 127.9,
125.6, 77.4, 68.6, 18.4(C-7).; HRMS(FAB) calcd for C20H17O6 (M+ + H) m/z 353.1025,
found 353.1023.; Anal. Calcd for C20H16O6: C, 68.18; H, 4.58. Found: C, 68.10; H, 4.61.
O
BzO
O
OBz
Compd. 7
3,5-Di-O-benzoyl-4,7-dideoxy-L-arabino-hexono-2,6-lactone (7)
Pd/C (10%, 400mg) was added to a solution of 6 (6.1g, 17.31mmol) in EtOAc (300mL).
The mixture was hydrogenated for 3h, and filtered through Celite. Concentration in
vacuo and purification by flash column chromatography (Toluene / EtOAc, 10:1, v/v)
afforded compound 7 (5.2g, 85%).
7; a white solid, Rf = 0.45 (Toluene / EtOAc, 10:1, v/v); [α]D21= +18.4 (c 1.0, CHCl3);
FT-IR (film) vmax 3031, 2982, 2939, 1724, 1601, 1452, 1383, 1273, 1114, 1070, 1028
cm-1; 1H NMR (250 MHz, CDCl3) δ 8.11-7.43 (m, 10H, aromatic H), 5.90 (dd, 1H, J =
7.6, 12.0 Hz, H-3), 5.30-5.25 (m, 1H, H-5), 4.87-4.77 (m, 1H, H-6), 2.78-2.52 (m, 2H,
14
H-4eq,4ax), 1.58 (d, 3H, J = 6.5 Hz, H-7); 13C NMR (62.9 MHz, CDCl3) δ 168.0(C2), 165.5(2), 133.9, 133.8, 130.2(2), 129.9(2), 129.1, 129.0, 128.8(2), 128.6(2), 76.9,
70.5, 65.0, 30.2(C-4), 19.3(C-7).; HRMS(FAB) calcd for C20H19O6 (M+ + H) m/z
355.1182, found 355.1178.; Anal. Calcd for C20H18O6: C, 67.79; H, 5.12. Found: C,
67.70; H, 5.26.
OH
BzO
O
OBz
Compd. 8
3,5-Di-O-benzoyl-4,7-dideoxy-L-arabino-hexopyranose (8)
To a pre-cooled 1M BH3-THF (65mL) at 0°C, 2M 2,3-dimethyl-2-butene (65mL) was
added slowly and stirred for 2 h. to give 0.5M Disiamylborane (127 mL). Compound 7
(5g, 14.11mmol) in dry THF (15mL) was added and stirred for 20 h. at r. t. After H2O
(3mL) was added, stirred for 30 min. and cooled to 0°C. 30% H2O2 (15mL) was added
and pH of the solution was adjusted to 7-8 by addition of 3N NaOH. The Concentration
in vacuo gave crude product which was dissolved in CH2Cl2 (100mL) and washed by
H2O (50mL) and dried by anhydrous MgSO4. Concentration in vacuo and purification
by flash column chromatography (Toluene / EtOAc, 10:1, v/v) afforded compound 8
(4.72g, 93.8%, α:β = 4.6:1).
8α; a colorless syrup, Rf = 0.23 (Toluene / EtOAc, 10:1, v/v); [α]D24= +51.4 (c 1.0,
CHCl3); FT-IR (film) vmax 3448, 3065, 3027, 2979, 1720, 1601, 1452, 1270, 1112, 1095,
15
1068, 1025 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.15-7.43 (m, 10H, aromatic H), 5.29 (s,
1H, H-2), 5.25-5.15 (m, 2H, H-2, H-5), 4.39-4.28 (m, 1H, H-6), 3.51 (d, 1H, J = 3.6
Hz, -OH), 2.44 (td, 1H, J = 3.8, 13.5 Hz, H-4eq), .2.29 (ddd, 1H, J = 3.1, 11.0, 13.7 Hz,
H-4ax), 1.30 (d, 3H, J = 6.2 Hz, H-7); 13C NMR (62.9 MHz, CDCl3) δ 166.0, 165.8,
133.5, 133.4, 130.0(3), 129.8(3), 128.6(4), 91.1(C-2, α), 71.0(C-3), 70.7(C-5), 67.0(C6), 29.2(C-4), 18.0(C-7).; HRMS(FAB) calcd for C20H21O6 (M+ + H) m/z 357.1338,
found 357.1334.; Anal. Calcd for C20H20O6: C, 67.41; H, 5.66. Found: C, 67.44; H, 5.53.
OH
H
R
Compd. 10
(R)-(-)-7-Octen-2-ol (10)
To a cloudy solution of magnesium (571mg, 23.5mmol) in dry THF (3mL), 5-bromo-1pentene (2.8mL, 23.5mmol) in dry THF (20mL) was added dropwise for 30 min. The
mixture solution was refluxed for 3 h. at 60°C and cooled to r. t. To the cooled to -78°C
solution of (R)-(+)-1,2-Epoxypropane 9 (1.12mL, 16.0mmol) and CuBr (230mg,
1.6mmol in dry THF (23mL), the above prepared Grignard reagent of 1M 4pentenylmagnesium bromide (23mL, 23.5mmol ) was added, warmed to r. t.. and stirred
for 4 h. The reaction mixture was added by aq. NH4Cl (10mL), extracted by Et2O
(20mL×2). The organic layer was dried by anhydrous MgSO4. Concentration in vacuo
and purification by flash column chromatography (Et2O / n-Pentane, 10:1, v/v) afforded
compound 10 (1.3g, 65%).
16
10; a colorless liquid, Rf = 0.15 (Et2O / n-Pentane, 5:1, v/v); [α]D20= - 11.11 (c 4.30,
acetone); FT-IR (film) vmax 3357, 2969, 2930, 2858, 1641, 1460, 1416, 1374, 1305,
1122 cm-1; 1H NMR (250 MHz, CDCl3) δ 5.89-5.73 (m, 1H, H-2), 5.03-4.92 (m, 2H, H1), 3.80-3.78 (m, 1H, H-7), 2.07 (m, 2H, H-3), 1.43-1.39 (m, 6H, H-4, 5, 6), 1.18 (d, 3H,
J = 6.1 Hz, -CH3); 13C NMR (62.9 MHz, CDCl3) δ 138.9(C-2), 114.4(C-1), 68.0(C-7),
39.2(C-6), 33.8(C-3), 29.0(C-4), 25.3(C-5), 23.5(C-8).; Anal. Calcd for C8H16O: C,
74.94; H, 12.58. Found: C, 72.87; H, 12.35.
R
O
BzO
O
OBz
Compd. 11
(2R)-Oct-7-en-2-yl
-3,5-di-O-benzyl-4,7-dideoxy-α-L-arabino-hexopyranoside
(11)
To a solution of 8 (2.0g, 5.61mmol, 1eq), 10 (1.08g, 8.42mmol), and 4Å molecular
sieves (200mg) in dry CH2Cl2 (30mL) under N2 cooled to 0°C, BF3-Et2O (2.85mL,
16.8mmol, 4eq) was added slowly and the whole solution was stirred for 10 h. Et3N
(5mL) was added to complete the reaction and the reaction mixture was filtered.
Concentration in vacuo and purification by flash column chromatography (n-Hexane /
EtOAc, 5:1, v/v) afforded compound 11 (1.89g, 72%).
17
11; a colorless syrup, Rf = 0.55 (n-Hexane / EtOAc, 5:1, v/v); [α]D22= +0.9 (c 1.0,
CHCl3); FT-IR (film) vmax 3069, 2974, 2933, 2859, 1723, 1602, 1451, 1316, 1267, 1152,
1108, 1068, 1025 cm-1; 1H NMR (250 MHz, CDCl3) δ 8.14-7.42 (m, 10H, aromatic H),
5.93-5.76 (m, 1H), 5.26-5.16 (m, 2H, H-3, H-5), 5.07-5.00 (m, 3H, H-2), 4.20-4.09 (m,
1H, H-6), 3.85 (m, 1H), 2.48-2.41 (m, 1H, H-4eq), 2.28-2.17 (m, 1H, H-4ax), 2.11 (m,
2H), 1.68-1.37 (m, 6H), 1.30 (d, 3H, J = 6.2 Hz), 1.20 (d, 3H, J = 6.1 Hz);
13
C NMR
(62.9 MHz, CDCl3) δ 165.9, 165.7, 138.9, 133.3, 133.2, 129.9(3), 129.6(2), 128.5(4),
114.5, 93.8(C-2, α), 72.5, 71.3, 70.7, 67.0, 37.0, 33.8, 29.8, 28.8, 25.3, 19.2, 17.9.;
HRMS(FAB) calcd for C28H35O6 (M+ + H) m/z 467.2434, found 467.2438.; Anal. Calcd
for C28H34O6: C, 72.08; H, 7.35. Found: C, 72.01; H, 7.31.
R
OH
O
BzO
O
O
OBz
Compd. 12
(6R)-6-(3,5-Di-O-benzoyl-4,7-dideoxy-α-L-arabino-hexopyranosyl)
heptanoic
acid (12)
To a stirred solution of compound 11 (1.8g, 3.86mmol) in acetone (150mL), NaHCO3
(972mg, 11.57mmol) and KMnO4 (3g, 19.29mmol) was added. The solution was stirred
for 12 h. at room temperature and acidified by 10% HCl (20mL). The reaction mixture
was extracted by EtOAc (100mL×2), washed by brine (70mL) dried over MgSO4,
18
filtered and concentrated in vacuo to give the crude product. The crude product was
purified by chromatography on silica gel using n-Hexane / EtOAc (5:1, v/v) as eluant to
afford the compound 12 (1.51g, 87%).
12; a colorless syrup, Rf = 0.13 (Hexane / EtOAc, 5:1, v/v); [α]D22= -1.9 (c 1.0, CHCl3);
FT-IR (film) vmax 3063, 2973, 2935, 1721, 1602, 1451, 1316, 1267, 1109, 1068, 1025
cm-1; 1H NMR (250 MHz, CDCl3) δ 10.69 (bs, 1H, -OH), 8.14-7.42 (m, 10H, aromatic
H), 5.26-5.17 (m, 2H, H-3, H-5), 4.98 (s, 1H, H-2), 4.19-4.08 (m, 1H, H-6), 3.87 (m,
1H), 2.47-2.36 (m, 3H), 2.28-2.17 (m, 1H, H-4ax), 1.72-1.45 (m, 6H), 1.31 (d, 3H, J =
6.2 Hz), 1.21 (d, 3H, J = 6.0 Hz); 13C NMR (62.9 MHz, CDCl3) δ 179.8, 165.8, 165.7,
133.3, 133.2, 130.0, 129.9(2), 129.8, 129.7(2), 128.5(4), 93.8(C-2, α), 72.4, 71.2, 70.7,
67.1, 36.7, 34.0, 29.7, 25.2, 24.6, 19.1, 17.9.; HRMS(FAB) calcd for C27H33O8 (M+ +
H) m/z 485.2175, found 485.2165.; Anal. Calcd for C27H32O8: C, 66.93; H, 6.66. Found:
C, 67.04; H, 6.64.
R
OH
O
HO
O
O
OH
Compd. 1
(-)-6R-(3R, 5R-dihydroxy-6S-methyltetrahydro-pyran-2R-yloxy) heptanoic acid
(1)
19
To a stirred solution of compound 12 (472.9mg, 0.976mmol) in MeOH (20mL) was
added NaOMe (52.7mg, 0.976mmol) at 0C. The solution was warmed to r. t. and was
stirred for 12 h. After solvent was removed in vacuo, the resultant crude product was
dissolved in H2O (20mL) and washed with CH2Cl2 (20mL×5) to remove methyl
benzoate. The aqueous solution was neutralized by acidic Amberlite IR-120 (H+)
(500mg), filtered and freeze dried to give a thick crude oil. The crude oil was purified
by chromatography on silica gel using EtOAc / MeOH, (11:1, v/v) as eluant to afford
the compound 1 (234.6mg, 87%).
1 ; a colorless syrup, Rf = 0.43 (EtOAc / MeOH, 11:1, v/v); [α]D25 = - 81.0 (c 0.1,
MeOH); FT-IR (film) vmax 3391, 2969, 2933, 1712, 1452, 1379, 1244, 1126, 1103, 1042,
1031 cm-1; 1H NMR (500 MHz, CD3OD) δ 4.64 (s, 1H, H-2), 3.80-3.77 (m, 1H, H-6),
3.72-3.71 (m, 1H, H-3), 3.63-3.59 (m, 1H, H-6), 3.54-3.49 (m, 1H, H-5), 2.30 (t, 2H, J
= 7.5 Hz, H-2), 1.96-1.92 (m, 1H, H-4eq), 1.79-1.74 (m, 1H, H-4ax), 1.61 (m, 2H, H3), 1.56-1.50 (m, 2H, H-5), 1.47 (m, 2H, H-4), 1.21 (d, 3H, J = 6.5 Hz, H-7), 1.12 (d,
3H, J = 6.5 Hz, H-7);
13
C NMR (125.7 MHz, CD3OD) δ 177.7(C-1), 97.6(C-2, α),
72.4(C-6), 71.3(C-6), 70.1(C-3), 68.5(C-5), 38.2(C-5), 36.1(C-4), 35.0(C-2), 26.5(C3), 26.1(C-4), 19.4(C-7), 18.2(C-7); HRMS(FAB) calcd for C13H25O6 (M+ + H) m/z
277.1651, found 277.1652.; Anal. Calcd for C13H24O6: C, 56.51; H, 8.75. Found: C,
56.55; H, 8.80.