1
486
Helvetica Chimica Acta ± Vol. 87 (2004)
Experimental Part
General. Silica gel (200 ± 300 mesh) for column chromatography (CC) and silica gel GF254 for thin-layer
chromatography (TLC) were obtained from Qingdao Marine Chemical, Ltd., China. Melting points (m.p). are
uncorrected. Optical rotations: Horiba SEPA-300 automatic polarimeter. IR Spectra: Bio-Rad FTS-135
À1
1
13
spectrophotometer, KBr pellets; in cm
spectrometers (400/100 and 500/125 MHz, resp.); d in ppm rel. to Me
70 eV): VG Autospec3000 spectrometer; in m/z (rel. %). GC/MS: Finnigan 4510 apparatus; at 70 eV (EI);
.
H- and C-NMR Spectra: Bruker AM-400 and DRX-500
4
Si as internal standard, J in Hz. EI-MS
(
capillary column: 30 m  0.25 mm, packed with 5% phenyl- and 95% methylsilicon on HP-5, with He as carrier
gas, heating at 58/min in the range of 160 ± 2408.
Fungal Material. The fresh fruiting bodies of Paxillus panuoides were collected at Ailao Mountains, Yunnan
Province, P. R. China, in August 1998. A voucher specimen was deposited at the Herbarium of the Kunming
Institute of Botany, the Chinese Academy of Sciences, People×s Republic of China.
Extraction and Isolation. The air-dried and powdered fruiting bodies (50 g) of P. panuoides were extracted
with CHCl
3
/MeOH 1 :1 (3Â) at r.t. The combined org. phase was evaporated in vacuo to give a deep brown
gum. The crude extract was subjected to CC (SiO
2
; petroleum ether/acetone gradient). The fraction eluted with
petroleum ether/acetone 1:1 afforded pure 1 (6 mg). The fraction eluted with petroleum ether/acetone 6 :4 gave
pure 4 (26 mg).
2
,3-Dihydroxy-N-[(1S,2S,3R)-2,3-dihydroxy-1-(hydroxymethyl)heptadecyl]tetracosanamide
2
5
(
0
7
Paxillamide; 1). White amorphous powder. M.p. 160 ± 1628 (petroleum ether/acetone). [a] 12.3 (c
D
.01, pyridine). IR (KBr): 3350, 3218, 2920, 2854, 1620, 1546, 1468, 1358, 1276, 1132, 1109, 1070, 984, 904, 874,
1
13
22. H- and C- NMR: see the Table. EI-MS: 699 (1.5), 682 (5), 681 (4), 651 (6), 668 (3), 472(11), 442 (13), 425
(10), 400 (21), 383 (3), 376 (4), 375 (18), 355 (2), 337 (4), 325 (5), 307 (4), 306 (17), 286 (4), 268 (9), 250 (4),
2
39 (2), 227 (9), 186 (4), 160 (4), 147 (12), 131 (24), 125 (13), 118 (33), 111 (29), 97 (49), 83 (68), 69 (64), 55
(100). HR-EI-MS: 699.6358 (M , C42
H
85NO
6
; calc. 699.6377).
2-Hydroxy-N-[(1S,2S,3R)-2,3-dihydroxy-1-(hydroxymethyl)heptadecyl]tetracosanamide (4). White amor-
phous powder. M.p. 141 ± 1438 (petroleum ether/acetone). [a]
D
9.6 (c 0.22, pyridine). IR (KBr): 3340,
1
3
220, 2919, 2850, 2487, 2395, 1619, 1544, 1468, 1353, 1068, 1027, 723. H-NMR ((D
5
)pyridine): 5.12 (m); 4.62 (dd,
J 7.6, 4.0); 4.52 (dd, J 10.6, 4.5); 4.43 (dd, J 10.6, 5.2); 4.35 (dd, J 6.5, 4.0); 4.28 (m). EI-MS: 683 (2, M ),
6
6
65 (11), 651 (5), 456 (13), 439 (18), 409 (22), 384 (24), 357 (27), 339 (4), 320 (7), 227 (13). HR-EI-MS:
83.6407 (M , C42
H
85NO
5
; calc. 683.6427).
Methanolysis and Derivatization. a) Compound 1 (3 mg) was refluxed at 808 for 16 h in 82% aq. MeOH
1.2 ml) containing 0.9m HCl. The mixture was extracted repeatedly with petroleum ether, and the combined
(
org. layers were dried (Na
ether/AcOEt 9 :1 ! 6 :4) to afford methyl 2,3-dihydroxytetracosanoate. H-NMR (CDCl
2
SO
4
) and evaporated. The resulting residue was purified by CC (SiO
2
, petroleum
1
3
): 4.22 (dd, J 4.2,
5
.6); 3.84 (m); 3.83 (s); 1.47 (m); 1.25 (br. s), 0.88 (t, J 7.0). EI-MS: 414 (M ), 355 ([M À 59] ).
b) The aq. MeOH phase from the above reaction was neutralized with sat. Na
to dryness. The residue was taken up in Ac O/pyridine 1 :1 and heated at 708 for 90 min. The mixture was diluted
with H O, extracted with AcOEt, the org. phase was evaporated, and the residue was purified by CC (SiO
hexane/AcOEt 8 :2) to afford 3. H-NMR (CDCl
2 3
CO soln. and concentrated
2
2
2
;
1
3
): 5.97 (d, J 9.2); 5.10 (dd, J 8.5, 3.1); 4.93 (dt, J 9.8, 3.1);
.47 (m); 4.29 (dd, J 11.6, 4.3); 4.00 (dd, J 11.6, 3.1); 2.08 (s), 2.05 (s); 2.05 (s); 2.03 (s); 1.12 ± 1.70 (br. s); 0.88
4
(
t, J 6.1). EI-MS: 486 (1, [M 1] ), 426 (2, [M 1 À AcOH] ), 366 (9, [M 1 À 2AcOH] ), 305 (25, [M À
3
AcOH] ), 245 (0.5, [M 1 À 4AcOH] ).
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