Structure of a novel diterpenoid ester, fritillahupehin from bulbs of Fritillaria hupehensis Hsiao and K.C. Hsia

Article abstract of DOI:10.1016/S0367-326X(02)00085-0

A new diterpenoid ester, fritillahupehin (1) together with three known fatty acids: palmitic acid (2), lignoceric acid (3) and azelaic acid (4) were isolated from the bulbs of Fritillaria hupehensis Hsiao and K.C. Hsia. The structure of fritillahupehin has been established to be ent-kauran-16β-yl lignocerate by means of spectroscopic and chemical evidence. Compounds 2-4 were isolated from Fritillaria sp. for the first time.

Full text of DOI:10.1016/S0367-326X(02)00085-0

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Fitoterapia 73 2002 288᎐291
Structure of a novel diterpenoid ester,
fritillahupehin from bulbs of Fritillaria
hupehensis Hsiao and K.C. Hsia
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Hanli Ruana, Yonghui Zhanga, Jizhou Wua, ,
Shukai Denga, Handong Sunb, Tetsuro Fujitac
aFaculty of Pharmaceutical Sciences, Tongji Medical College of Huazhong Uni¨ersity of Science
and Technology, Wuhan 430030, PR China
bKunming Institute of Botany, Chinese Academy of Sciences, Kunming 650204, PR China
cFaculty of Pharmaceutical Sciences, Kyoto Uni¨ersity, Kydo 606-01, Japan
Received 5 December 2001; accepted in revised form 2 April 2002
Abstract
A new diterpenoid ester, fritillahupehin
palmitic acid 2 , lignoceric acid 3 and azelaic acid 4 were isolated from the bulbs of
Fritillaria hupehensis Hsiao and K.C. Hsia. The structure of fritillahupehin has been
established to be ent-kauran-16␤-yl lignocerate by means of spectroscopic and chemical
evidence. Compounds 2–4 were isolated from Fritillaria sp. for the first time. ᮊ 2002
Elsevier Science B.V. All rights reserved.
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together with three known fatty acids:
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Keywords: Fritillaria hupehensis; Diterpenoid ester; Fritillahupehin
1. Introduction
Fritillaria hupehensis is a liliaceous plant growing in the south-west district of
Hubei Province, China. Its bulbs have been recorded in the Pharmacopoeia of the
People’s Republic of China as a principal Chinese traditional medicine named
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‘Hubeibeimu’ 1 .
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Corresponding author.
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E-mail addresses: ruanhl@27.net H. Ruan , wujiz@tjmu.edu.cn J. Wu .
0367-326Xr02r$ - see front matter ᮊ 2002 Elsevier Science B.V. All rights reserved.
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PII: S 0 3 6 7 - 3 2 6 X 0 2 0 0 0 8 5 - 0

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We have previously reported the isolation of ten C-nor-D-homo steroidal alka-
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loids from the bulbs of this plant 2᎐8 . A number of the non-basic constituents
were also isolated: sitosterol, ent-kauran-16␤, 17-diol, ent-kauran-16␣, 17-diol,
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fritillebin C and fritillebin D 9,10 . In our continuing studies on the non-basic
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constituents, a novel diterpenoid ester, fritillahupehin 1 and three fatty acids:
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palmitic acid 2 , lignoceric acid 3 and azelaic acid 4 were isolated. The paper
describes the structural elucidation of compounds 1.
2. Experimental
2.1. General experimental procedures
Melting points were determined on X4 apparatus and are uncorrected. Optical
rotations were taken on WZZ-1digital polarimeter. IR spectra were taken on
Shimadzu IR-460 spectrometer. EI-MS spectra were measured on Auto-Spe mass
spectrometer. 1H-NMR and 13C-NMR data were recorded on Bruker-400 spec-
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trometer using CDCl3 as solvent. TLC: silica gel QingDao, China . Column
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chromatography: silica gel 100᎐200 mesh, QingDao, China .
2.2. Plant material
The bulbs of F. hupehensis Hsiao and K.C. Hsia were bought from HuBei
Institute of Chinese Materia Medeca and identified by PengDeTai, LiChuan
Institute of Chinese Materia Medeca, China.
2.3. Extraction and isolation
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The powdered bulbs 12 kg of F. hupehensis were extracted with EtOH. The
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extract 484.75 g was partitioned between n-hexane and water. The n-hexane
extract was fractionated by CC over silica gel, and eluted with n-hexane᎐EtOAc
containing increasing contents of EtOAc to get fraction II, 155 g. Part 86 g of
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fraction II was fractionated by CC over silica gel and eluted with petroleum᎐EtOAc
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to obtain: fritillahupehin 53 mg 1 ; palmitic acid 2 158.7 mg ; lignoceric acid
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69 mg 3 ; and azelaic acid 4 75.3 mg .
3. Results and discussion
The EtOH extract of F. hupehensis was partitioned between water and n-hexane.
The n-hexane extract was fractionated by column chromatography over silica gel to
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afford 1᎐4 . Fritillahupehin 1 , colorless needles mp 183᎐185 ЊC EtOAc
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D
y78.2Њ c 0.67, CHCl3 ; IR bands KBr : 1738, 1382, 1365, 720 cmy1; FAB-MS
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mrz:Mq 731 M y H q Gly . Found: C, 82.92; H, 12.24. Calc. for C44 H80O2: C,
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82.50; H, 12.50 .

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H. Ruan et al. rFitoterapia 73 2002 288᎐291
Fig. 1. Fritillahupehin.
Alkaline hydrolysis of fritillahupehin with 5% NaOH᎐MeOH yielded two com-
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1
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pounds. The C-NMR 11 and H-NMR 12 spectra of the neutral component
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were in accordance with those of ent-kauran-16␤-ol 16␣-hydroxy-ent-kaurane
which had been isolated previously from Xylopia aethiopica 13 .
The acidic component was identified as lignoceric 3 from spectral parameters
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and direct comparison with the authentic sample.
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From the evidence described above, the structure of fritrillahupehin 1 was
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established as ent-kauran-16-␤-yl lignocerate Fig. 1 .
The other compounds recovered from chromatography were identified as palmitic
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2 , lignoceric 3 and azelaic acid 4 .
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