1052 Journal of Natural Products, 2010, Vol. 73, No. 6
Arnone et al.
Isolation of 4 and 5 from Cultures of White 176-ICRM and Black
198-ICRM Strains. The crude residues (0.9 g 176-ICRM; 0.85 g 198-
ICRM) were purified by column chromatography on silica gel with a
stepwise elution with CH2Cl2/i-PrOH from 30:1 to 1:1. The fractions
obtained were further purified on preparative plates (PLC) in reverse
phase (RP-18): metabolite 4 was isolated from both strains of fungus
C. lunata, using H2O/MeCN (2:1) as eluent to give 3 mg (0.3%) from
white cultures (176-ICRM) and 4 mg (0.4%) from black cultures (198-
ICRM). Compound 5 (13 mg, 1.4%) was obtained only from cultures
of the white strain using H2O/MeCN (1:1) as eluent; unreacted 10-
DAB (1) was recovered (260 mg, 22%).
12 Union Road, Cambridge CB21EZ, UK (fax: +44-(0)12233336033
Acknowledgment. We are indebted to Prof. G. Appendino for
helpful discussions, Dr. F. Zunino (Istituto Nazionale dei Tumori,
Milano) for the cytotoxicity tests, and Indena Spa for financial support.
1
Supporting Information Available: NOEs (Table 4) and H and
13C NMR spectra for the new compound 4. This material is available
Compound 4: white solid, mp 190 °C; [R]D -36 (c 0.2, MeOH);
UV (EtOH) λmax 203, 230, and 273sh (ε 15.300, 12.400, and 1200);
ESIMS m/z 523 [M + Na]+, 501 [M + H]+, and 482 [M + H - 18]+;
FABMS m/z 501 [M + H]+; HREIMS 500.2038 (calcd for C27H32O9,
References and Notes
(1) Kingston, D. G. I. Phytochemistry 2007, 68, 1844–1854.
(2) Dai, J.; Qu, R.; Zou, J.-H.; Chen, X. Tetrahedron 2008, 64, 8102–
8116.
(3) Sun, D.-A.; Nikolakakis, A.; Sauriol, F.; Mamer, O.; Zamir, L. O.
Bioorg. Med. Chem. 2001, 9, 1985–1992.
(4) Jennewein, S.; Croteau, R. Appl. Microbiol. Biotechnol. 2001, 57, 13–
19.
(5) Arnone, A.; Bava, A.; Alemani, S.; Nasini, G.; Bombardelli, E.;
Fontana, G. J. Mol. Catal. B: Enzym. 2006, 42, 95–98.
(6) Arnone, A.; Bava, A.; Fronza, G.; Nasini, G. J. Nat. Prod. 2009, 72,
2000–2004.
1
500.2046); H and 13C NMR data are in Tables 1 and 2.
Compound 5: white solid, mp 165 °C; [R]D -30 (c 0.1, MeOH);
ESIMS m/z 565 [M + Na]+, 543 [M + H]+, and 524 [M + H - 18]+;
ESIMS, negative ion m/z 541 [M - H]+; HREIMS 542.2127 (calcd
for C29H34O10, 542.2148); 1H and 13C NMR data are in Tables 1 and 2.
Cytotoxicity Bioassay. Compounds 4, 5, 6, and 10-Dab (1) were
tested for their cytotoxicity against the non-small-cell lung tumor cell
line H 460 [IC50 (µM): 102.8, 68, 60, and 3.1, respectively].
(7) This adduct crystallizes in the orthorhombic system, space group
P212121, with cell parameters a ) 8.383(1) Å, b ) 10.846(1) Å, c )
31.148(2) Å, V )2805.9(2) Å3, Z ) 4, Dc ) 1.320 g cm-3, F(000) )
1188. In the crystal the molecules are kept together, forming zig-zag
chains, elongated parallel to the c axis, by C5-H · · · O4 hydrogen
bonding. Full data (excluding structure factors) of this crystal structure
have been deposited as supplementary publication No. CCDC 769838.
(8) Vander Velde, D. G.; Georg, G. I.; Gollapudi, S. R.; Jampani, H. B.;
Liang, X.-Z.; Mitscher, L. A.; Ye, Q.-M. J. Nat. Prod. 1994, 57, 862–
867.
(9) Zamir, L. O.; Balachandran, S.; Zheng, Y. F.; Nedea, M. E.; Caron,
G.; Nikolakakis, A.; Vishwakarma, R. A.; Sauriol, F.; Mamer, O.
Tetrahedron 1997, 47, 15991–16008.
(10) Shen, Y.-C.; Wang, S.-S.; Pan, Y.-L.; Lo, K.-L.; Chakraborty, R.;
Chien, C.-T.; Kuo, Y. H.; Lin, Y.-C. J. Nat. Prod. 2002, 65, 1848–
1852.
(11) Appendino, G.; Noncovich, A.; Bettoni, P.; Dambruoso, P.; Sterner,
O.; Fontana, G.; Bombardelli, E. Eur. J. Org. Chem. 2003, 4422–
4431.
(12) Appendino, G.; Jakupovic, J.; Varese, M.; Bombardelli, E. Tetrahedron
Lett. 1996, 37, 727–730.
(13) Pattel, R. N. Food Technol. Biotechnol. 2004, 42, 305–325.
(14) Li, Y.-C.; Tao, W.-Y. Cell Biol. Int. 2009, 33, 106–112.
(15) Sheldrick, G. M. SADABS; University of Go¨ttingen, Germany, 1996.
(16) Altomare, A.; Burla, M. C.; Camalli, M.; Cascarano, G. L.; Giaco-
vazzo, G.; Guagliardi, A.; Moliterni, A. G. G.; Polidori, G. P.; Spagna,
J. Appl. Crystallogr. 1999, 32, 115–119.
Crystal Structure Determination of 4. The compound crystallizes
in the orthorhombic system, space group P212121, with unit cell
parameters a ) 8.362(1) Å, b ) 10.869(1) Å, c ) 27.818(2) Å,, V )
2528.3(2) Å3, Z ) 4, Dc ) 1.362 g cm-3, F(000) ) 1104. A crystal
suitable for X-ray analysis, with dimensions of 0.4× 0.2 × 0.15 mm,
was obtained upon slow recrystallization from acetone/water.
Intensity data were collected, at room temperature, on a Bruker AXS
SMART Apex single-crystal diffractometer with a 5 cm crystal-to-
detector distance and graphite-monochromatized Mo KR radiation
(λ ) 0.71069 Å) at 50 kV and 30 mA. Unit-cell dimensions were
calculated from least-squares refinement of 5017 reflections in the 2θ
range 4.8-45.3°. A total of 47 782 reflections was collected in the θ
range 2.4-27.0°, corresponding to 5522 unique reflections (Rint
)
0.033). Raw intensity data were corrected for absorption using the
SADABS v. 2.10 program.15
The structure was solved by the direct method using the SIR97
program,16 which revealed the position of all non H-atoms. The
refinement was carried out on F2 by a full-matrix least-squares
procedure with SHELXL97 for 384 parameters, with anisotropic
temperature factors for non-H atoms.17 The final stage converged to R
) 0.0467 (Rw ) 0.1192) for 5080 observed reflections (with I g 2σ(I)),
and R ) 0.0510 for all reflections. The hydroxy and the tertiary H
atoms were freely and isotropically refined; all other H atoms were
placed in geometrically calculated positions and refined in a riding
model.
(17) Sheldrick, G. M. SHELXL-97, Program for the Refinement of Crystal
Structures; University of Go¨ttingen: Germany, 1997.
Full data (excluding structure factors) of the crystal structure have
been deposited as supplementary publication No. CCDC 755642. Copies
of the data can be obtained, free of charge, on application to the CCDC,
NP900765Y