H. Kanatani et al. / Tetrahedron Letters 46 (2005) 8467–8470
8469
ella sp. (TM-I1104). We also thank Dr. Masahide
Tanaka for the offer of synthetic ( )-2.
References and notes
1. (a) Taguchi, H.; Ikeya, Y. Chem. Pharm. Bull. 1975, 23,
3296–3298; (b) Ikeya, Y.; Taguchi, H.; Yoshioka, I.;
Kobayashi, H. Chem. Pharm. Bull. 1979, 27, 1383–
1394.
2. (a) Maeda, S.; Sudo, K.; Miyamoto, Y.; Takeda, S.;
Shinbo, M.; Aburada, M.; Ikeya, Y.; Taguchi, H.;
Harada, M. Yakugaku Zasshi 1982, 102, 579–588; (b)
Kiso, Y.; Tohkin, M.; Hikino, H.; Ikeya, Y.; Taguchi, H.
Planta Med. 1985, 51, 331–334; (c) Maeda, S.; Takeda, S.;
Miyamoto, Y.; Aburada, M.; Harada, M. Jpn. J. Phar-
macol. 1985, 38, 347–353; (d) Yamada, S.; Murawaki, Y.;
Kawasaki, H. Biochem. Pharmacol. 1993, 46, 1081–1085;
(e) Shiota, G.; Yamada, S.; Kawasaki, H. Res. Commun.
Mol. Path. 1996, 94, 141–146.
3. (a) Ohkura, Y.; Mizoguchi, Y.; Sakagami, Y.; Kobayashi,
K.; Yamamoto, S.; Morisawa, S.; Takeda, S.; Aburada,
M. Jpn. J. Pharmacol. 1987, 44, 179–185; (b) Cyong, J. C.;
Ki, M. S.; Iijima, K.; Kobayashi, T.; Furuya, M. Am. J.
Chin. Med. 2000, 28, 351–360.
4. (a) Nomura, M.; Nakachiyama, M.; Hida, T.; Ohtaki, Y.;
Sudo, K.; Aizawa, T.; Aburada, M.; Miyamoto, K.
Cancer Lett. 1994, 76, 11–18; (b) Ohtaki, T.; Nomura,
M.; Hida, T.; Miyamoto, K.; Kanitani, M.; Aizawa, T.;
Aburada, M. Biol. Pharm. Bull. 1994, 17, 808–814; (c)
Nomura, M.; Ohtaki, Y.; Hida, T.; Aizawa, T.; Wakita,
H.; Miyamoto, K. Anticancer Res. 1994, 14, 1967–
1972.
5. (a) Takeda, S.; Kase, Y.; Arai, I.; Hasegawa, M.;
Sekiguchi, Y.; Funo, S.; Aburada, M.; Hosoya, E. Nippon
Yakurigaku Zasshi 1986, 88, 321–330; (b) Takeda, S.;
Kase, Y.; Arai, I.; Ohkura, Y.; Hasegawa, M.; Sekiguchi,
Y.; Tatsugi, A.; Funo, S.; Aburada, M.; Hosoya, E.
Nippon Yakurigaku Zasshi 1987, 90, 51–65; (c) Kubo, S.;
Ohkura, Y.; Mizoguchi, Y.; Matsui-Yuasa, I.; Otani, S.;
Morisawa, S.; Kinoshita, H.; Takeda, S.; Aburada, M.;
Hosoya, E. Planta Med. 1992, 58, 489–492; (d) Hirotani,
Y.; Kurokawa, N.; Takashima, N.; Sawada, M.; Iguchi,
K.; Yanaihara, N.; Iwasaki, M.; Aburada, M.; Arakawa,
Y.; Yanaihara, C. Biomed. Res. 1995, 16, 43–50.
6. (a) Tanaka, M.; Mukaiyama, C.; Mitsuhashi, H.; Wak-
amatsu, T. Tetrahedron Lett. 1992, 33, 4165–4168; (b)
Tanaka, M.; Mukaiyama, C.; Mitsuhashi, H.; Maruno,
M.; Wakamatsu, T. J. Org. Chem. 1995, 60, 4339–
4352.
Figure 1. HPLC profile of culture broth11 by Mortierella sp. (TM-
I1104) analytical condition; column, YMC A-312 (C18, 150 · 6 mm);
flow rate, 1.0ml/min; wavelength, 254 nm; mobile phase, H 2O–
MeOH–CH3CN–tetrahydrofuran (40:15:6:3, v/v). The retention times
of 2 and 3 were 11.7 and 23.6 min, respectively. 4, 13-norschizandrin;
5, 2-norschizandrin; 6, 3-norschizandrin.
separated by preparative HPLC to afford the produced
25
3 (amorphous powder, ½aꢀD +93.6), which was con-
1
13
firmed by comparison of the H and C NMR data12
with those of natural 3. The circular dichroism (CD)
spectrum of produced 3 showed a positive Cotton
effect at 249 nm ([h] +80,442) and a negative Cotton
effect at 215 nm ([h] ꢁ93,821), indicating that the prod-
uct possesses an R-biphenyl configuration.7 On the
other hand, the CD spectrum of the recovered substrate
shows a positive Cotton effect at 215 nm ([h] +103,109)
and a negative Cotton effect at 249 nm ([h] ꢁ94,055),
indicating that the recovered substrate possesses an S-
biphenyl configuration,7 thus to be (ꢁ)-2. The optical
purity of produced (+)-3 was determined to be 96%
ee by the following method. Produced (+)-3 was first
derived to (+)-213 by O-methylation with methyl iodide,
then subjected to HPLC analysis using a chiral
column.14
7. Ikeya, Y.; Taguchi, H.; Yoshioka, I.; Kobayashi, H.
Chem. Pharm. Bull. 1979, 27, 1383–1394.
8. Kanatani, H.; Sakakibara, C.; Tanaka, M.; Niitsu, K.;
Ikeya, Y.; Wakamatsu, T.; Maruno, M. Biosci. Biotechnol.
Biochem. 1994, 58, 1054–1057.
In conclusion, we have discovered a competent fungus,
Mortierella sp. (TM-I1104), which converted 2 into 3
in 80% yield and 91% regioselectivity. This fact signifies
that the microbial 12-O-demethylation of isolated natu-
ral 2 by TM-I1104 theoretically permits the total synthe-
sis of 1 in 52% overall yield. Moreover, the microbial
12-O-demethylation of ( )-2 by TM-I1104 produced 3
in 96% ee. This result indicates that synthetic ( )-2 is
also available as a starting material for the total synthe-
sis of 1.
9. Culture medium for screening contains 0.5% potato
dextrose broth, 2% glucose, 0.5% yeast extract, 0.5%
sodium chloride, 0.5% potassium phosphate and pH 7.0.
10. The analysis of each culture broth by HPLC was done
under the following conditions: column, YMC A-312 (C18,
150 · 6 mm); mobile phase, H2O–MeOH–CH3CN–tetra-
hydrofuran (40:15:16:3); flow rate, 1 ml/min; detection,
wavelength, 254 nm. The retention time of 2 and 3 were
11.7 and 23.6 min, respectively.
11. The incubation in the optimum condition was performed
in the culture medium containing 4% glucose, 1% peptone,
0.07% KH2PO4, 0.05% MgSO4Æ7H2O, pH 5.5 at 25 ꢁC for
2 weeks.
Acknowledgements
We are grateful to Dr. Tatsuo Yokoyama (Institute for
Fermentation, Osaka) for the identification of Mortier-