Insecticidal and α-glucosidase inhibitory activities of chemical constituents from Viburnum fordiae Hance

Article abstract of DOI:10.1080/14786419.2018.1466130

The ethanolic extract of the stems of Viburnum fordiae Hance showed insecticidal and α-glucosidase inhibitory activities and then was fractionated by bioactivity-guided fractionation to obtain a rare C₁₃-norisoprenoid (1), together with a new phenolic glycoside (2), and seven known compounds, alangionoside C (3), pisumionoside (4), koaburaside (5), 3,5-dimethoxy-benzyl alcohol 4-O-β-d-glucopyranoside (6), 3,4,5-trimethoxybenzyl-β-d-glucopyranoside (7), arbutin (8), and salidroside (9). The previously undescribed compounds were elucidated as (3R,9R)-3-hydroxy-7,8-didehydro-β-ionyl 9-O-α-d-arabinopyranosyl-(1→6)-β-d-glucopyranoside (1) and 2-(4-O-β-d-glucopyranosyl)syringylpropane-1,3-diol (2) by spectroscopic data (¹H and ¹³C NMR, HSQC, HMBC, ¹H-¹H COSY, HSQC-TOCSY, HRESIMS, IR and ORD) and chemical methods. Compound 1 showed potent insecticidal effect against Mythimna separata with LD₅₀ value of 140 μg g^?1. Compounds 2, 5, 6, 8 and 9 showed varying α-glucosidase inhibitory activity with IC₅₀ values ranging from 148.2 to 230.9 μM.

Full text of DOI:10.1080/14786419.2018.1466130

Natural Product Research
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Insecticidal and α-glucosidase inhibitory activities
of chemical constituents from Viburnum fordiae
Hance
Jian-Hua Shao, Jia Chen, Chun-Chao Zhao, Jie Shen, Wen-Yan Liu, Wen-Yan
Gu & Ke-Huan Li
To cite this article: Jian-Hua Shao, Jia Chen, Chun-Chao Zhao, Jie Shen, Wen-Yan Liu, Wen-Yan
Gu & Ke-Huan Li (2018): Insecticidal and α-glucosidase inhibitory activities of chemical constituents
from Viburnum fordiae Hance, Natural Product Research, DOI: 10.1080/14786419.2018.1466130
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Natural Product research, 2018
https://doi.org/10.1080/14786419.2018.1466130
Insecticidal and α-glucosidase inhibitory activities of chemical
constituents from Viburnum fordiae Hance
Jian-Hua Shaoa , Jia Chena, Chun-Chao Zhaoa,b , Jie Shenb, Wen-Yan Liub,
Wen-Yan Gua and Ke-Huan Lia
aJiangsu Key laboratory of crop Genetics and Physiology/co-Innovation center for Modern Production
b
technology of Grain crops, Yangzhou university, Yangzhou, china; Joint International research laboratory
of agriculture & agri-Product safety of Ministry of education of china, Yangzhou university, Yangzhou, china
ABSTRACT
ARTICLE HISTORY
received 8 February 2018
accepted 14 april 2018
The ethanolic extract of the stems of Viburnum fordiae Hance showed
insecticidal and α-glucosidase inhibitory activities and then was
fractionated by bioactivity-guided fractionation to obtain a rare
C13-norisoprenoid (1), together with a new phenolic glycoside (2),
and seven known compounds, alangionoside C (3), pisumionoside
(4), koaburaside (5), 3,5-dimethoxy-benzyl alcohol 4-O-β-d-
KEYWORDS
Viburnum fordiae hance;
chemical constituents;
insecticidal activity;
α-glucosidase inhibitory
activity
glucopyranoside (6), 3,4,5-trimethoxybenzyl-β- -glucopyranoside (7),
d
arbutin (8), and salidroside (9). The previously undescribed compounds
were elucidated as (3R,9R)-3-hydroxy-7,8-didehydro-β-ionyl 9-O-α-
d-arabinopyranosyl-(1→6)-β-d-glucopyranoside (1) and 2-(4-O-β-d-
glucopyranosyl)syringylpropane-1,3-diol (2) by spectroscopic data (1H
and 13C NMR, HSQC, HMBC, 1H-1H COSY, HSQC-TOCSY, HRESIMS, IR and
ORD) and chemical methods. Compound 1 showed potent insecticidal
effect against Mythimna separata with LD50 value of 140 μg g−1
.
Compounds 2, 5, 6, 8 and 9 showed varying α-glucosidase inhibitory
activity with IC50 values ranging from 148.2 to 230.9 μM.
1. Introduction
Many species of Viburnum, belonging to the family Adoxaceae (formerly Caprifoliaceae),
have attracted attention as garden or landscape plants because of their showy flowers and
CONTACT chun-chao Zhao
cczhao@yzu.edu.cn; Jia chen
chartishchen@hotmail.com
supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2018.1466130.
© 2018 Informa uK limited, trading as taylor & Francis Group

2
J.-H. SHAO ET AL.
Figure 1. the structures of compounds 1–9.
berries. More importantly, they have notable biological activities that can improve human
health and treat various diseases (El-Gamal 2008; Seeram 2008; Saltan et al. 2016). Previous
phytochemical and pharmacological investigations on Viburnum species clearly indicated
that these plants contain monoterpenoids, sesquiterpenoids, diterpenoids, triterpenoids,
iridoids, lignans, flavonoids and other phenolic compounds, some of which possessed anti-
tumor, antimicrobial, antioxidant, antihyperglycemic, anti-inflammatory and neuroprotective
activities (Wang et al. 2010; In et al. 2015). Viburnum fordiae Hance is a small tree widely
distributed in the south of China. As a common traditional Chinese medicine, its stems have
been used for the treatment of rheumatic arthralgia and allergic dermatitis (Zhonghuabencao
Editorial Board 1999). The crimson fruits of V. fordiae, ripening in autumn, have also been
reported as a promising functional food (Shao et al. 1992). To the best of our knowledge,
there were minimal work on the chemical constituents and biological activities focusing on
V. fordiae. As part of our continuing search for bioactive constituents from the genus Viburnum
(Shao et al. 2018), we investigated the ethanol extract from the stems of V. fordiae. Nine
compounds were isolated from its n-butanol fraction (Figure 1). Among them, two previously
undescribed compounds (1, 2) have been unambiguously determined by extensive spec-
troscopic methods, including 1D (1H and 13C) and 2D NMR (HSQC, HMBC, 1H-1H COSY, and
HSQC-TOCSY), IR and HRESIMS, and chemical transformations. The insecticidal and α-gluco-
sidase inhibitory activities of all isolated secondary metabolites were assayed. Herein, we
describe the isolation, structural elucidation and evaluation of insecticidal and α-glucosidase
inhibitory activities of these compounds.

NATURAL PRODUCT RESEARCH
3
2. Results and discussion
Compound 1 was isolated and purified as white amorphous power. Its positive HRESIMS
spectrum exhibited a sodium adduct molecular ion peak [M + Na]+ at m/z 525.2321 (Calcd
525.2306), corresponding to the molecular formula C24H38O11 with 6° of unsaturation. The
IR spectrum indicated the presence of alkyne (vmax 2209 cm−1) and alkene (vmax 1623 cm−1)
1
groups. The H NMR spectrum of 1 (Table S1, Supplementary material) displayed proton
signals corresponding to two oxygenated methine groups at δH 3.72 (1H, m, H-3) and 4.78
(1H, q, J = 6.5 Hz, H-9), two methylene moieties at δH 1.26 (1H, t, J = 12.1 Hz, H-2ax), 1.69 (1H,
ddd, J = 12.1, 3.7, 2.1 Hz, H-2 eq), 1.90 (1H, dd, J = 17.5, 9.7 Hz, H-4ax) and 2.25 (1H, dd,
J = 17.5, 5.6 Hz, H-4 eq), four methyl residues at δH 1.05 (3H, s, H-11), 1.09 (3H, s, H-12), 1.40
(3H, d, J = 6.5 Hz, H-10) and 1.80 (3H, s, H-13). Combining with HSQC spectrum of 1, the 13
C
NMR spectrum showed 13 signals of aglycone, assigned to four methyl groups at δC 22.2
(C-13), 22.5 (C-10), 28.4 (C-11) and 30.3 (C-12), two methylene moieties at δC 41.1 (C-4) and
46.4 (C-2), two oxygenated methine groups at δC 62.5 (C-3) and 62.6 (C-9) and five quaternary
carbons at δC 35.9 (C-1), 83.1 (C-7), 92.7 (C-8), 122.5 (C-6) and 138.3 (C-5). The above data,
together with 6° of unsaturation, implied that compound 1 should be a C13-norisoprenoid
(Cutillo et al. 2005; Picerno et al. 2008). Acid hydrolysis of 1 resulted in liberating d-glucose
and d-arabinose, which were analysed by TLC, ORD and GC, and the configuration of the
pentose was determined by Hara’s method (Hara et al. 1987; Seo et al. 2002; Sun et al. 2016).
The d-glucose and d-arabinose were confirmed to have β- and α-linkages by the coupling
constants of anomeric protons at δH 4.43 (1H, d, J = 7.8 Hz, H-1′) and 4.17 (1H, d, J = 6.2 Hz,
H-1″), respectively (Pei et al. 2011). In the HMBC spectrum of 1 (Figure S16, Supplementary
material), key correlations from H-10 (δH 1.40) to C-9 (δC 62.6) and C-8 (δC 92.7), from H-9 (δH
4.78) to C-8 and C-7 (δC 83.1), from H-2 (δH 1.26, 1.69) and H-4 (δH 1.90, 2.25) to C-3 (δC 62.5),
and from H-13 (δH 1.80) to C-5 (δC 138.3) and C-6 (δC 122.5) indicated that two oxygenated
groups, an alkynyl moiety, and an alkenyl residue were substituted at C-9, C-3, C-7 and C-5
positions, respectively. Also, the correlations of H-1′ (δH 4.43) with C-9 (δC 62.6) and H-1″ (δH
4.17) with C-6′ (δC 64.8) indicated that the linkage points of the glucose and arabinose units
were at C-9 and C-6′, respectively. Based on the above data and comprehensive 2D NMR
experiments (1H-1H COSY, HSQC-TOCSY and HMBC), the structure of 1 was unambiguously
assigned as shown in Figure 1. According to the literature, in the 13C NMR spectra of 1 in D2O
(Table S1, Supplementary material), the chemical shift (δC 68.2) of C-9 indicated that the
absolute configuration of C-9 was R (Yamano et al. 2002). The relative stereochemistry at C-3
in the aglycone group of 1 was identified by a large coupling constant of H-4ax (J = 9.7 Hz).
This implied that H-3 must be in the axial position (Yue et al. 2012). C13-norisoprenoids are
biosynthetically formed in nature from the hydrolytic breakdown of complex secondary
metabolites derived from carotenoids which have R-configuration for C-3 (Yamano et al.
2000; Baumes et al. 2002;Yamano et al. 2002). Thus, compound 1 was determined as (3R,9R)-
3-hydroxy-7,8-didehydro-β-ionyl 9-O-α-d-arabinopyranosyl-(1→6)-β-d-glucopyranoside.
Compound 2, white amorphous powder, had the molecular formula C17H26O10 as deduced
by analysis of positive HRESIMS (m/z 413.1442 [M + Na]+, Calcd 413.1418). Hydrolysis of 2
with β-glucosidase produced an aglycone and β-d-glucose, which was identified by the
positive optical rotation[ꢀ]2D3 = +48.6 (c 0.2, H2O) (Jiang et al. 2015). The 1H NMR spectrum
of 2 (Table S2, Supplementary material) indicated the presence of a 1,3,4,5-tetrasubstituted
benzene ring residue [δH 6.52 (2H, s, H-2′, 6′)], two methoxyl groups [δH 3.74 (6H, s, -OCH3-3′,

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J.-H. SHAO ET AL.
5′)], and a propane chain [two methylene moieties at δH 3.58 (2H, m, H-1, 3), 3.67 (2H, m, H-1,
3) and a methine group at δH 2.72 (1H, m, H-2)]. Combining with HSQC spectrum of 2, this
aliphatic part was further confirmed in the 13C NMR spectrum by a methine carbon signal
at δC 50.7 (C-2) and a methylene peak at δC 62.7 (C-1 and C-3). Considering only 7 carbon
resonances corresponding to the aglycone in the 13C NMR spectrum together with the above
molecular formula, it was obvious that the aglycone of compound 2 should possess a sym-
metrical structure. The structure of 2 was further determined by HMBC experiment. Key
HMBC correlations from H-1″ (δH 4.83) to C-4′ (δC 133.2), from H-2 (δH 2.72) to C-1′ (δC 137.9),
C-2′ (δC 106.7) and C-6′ (δC 106.7), and from H-1 (δH 3.58, 3.67) and H-3 (δH 3.58, 3.67) to C-1′
showed the linkage points of glucosyl unit and syringyl group, respectively. Consequently,
compound 2 was identified as 2-(4-O-β-d-glucopyranosyl)syringylpropane-1,3-diol.
Compounds 3–9, by comparing the physical and spectral data with literature values
(Experimental Section, Supplementary material), were determined as alangionoside C (3),
pisumionoside (4), koaburaside (5), 3,5-dimethoxy-benzyl alcohol 4-O-β- -glucopyranoside
d
(6), 3,4,5-trimethoxybenzyl-β-d-glucopyranoside (7), arbutin (8) and salidroside (9).
The n-butanol soluble fraction of V. fordiae was initially assayed for its insecticidal and
α-glucosidase inhibitory activities (Experimental Section, Supplementary material). The
obtained data of bioassay revealed that the n-butanol extract showed the moderate insec-
ticidal activity against Mythimna separata Walker with LD50 value of 297 μg g−1 and α-gluco-
sidase inhibitory activity with IC50 value of 258 μg mL−1. Subsequently, the n-butanol extract
was isolated and purified by chromatographic methods to give nine compounds (1–9). These
compounds were further tested for insecticidal and α-glucosidase inhibitory activities. New
compound 1 showed potent insecticidal effects against M. separata with LD50 value of
140 μg g−1 in comparison with isonimbinolide (124 μg g−1) and celangulin-V (312 μg g−1),
while compounds 3 and 4 were inactive. This might be attributed to the existence of an
alkynyl unit at C-7 of 1. Among the tested phenolic compounds, 2, 5, 6, 8 and 9 exhibited
potent intestinal α-glucosidase inhibitory activity with IC50 values ranging from 148.2 to
230.9 μM in comparison to the positive control acarbose (IC50 = 220.4 μM) (Figure S18,
Supplementary material), while 7 did not show inhibitory effect. This result preliminarily
indicated that the free hydroxyl group in aglycone played a key role in exerting inhibition
of α-glucosidase. Postprandial hyperglycemia (PPHG), treated by α-glucosidase inhibitors
via delaying the digestion of carbohydrates in the intestines, is involved in the development
of type 2 diabetes (Yao et al. 2010). Obviously, the obtained data of the bioactivity assays
indicated that V. fordiae Hance containing these constituents could be developed as drugs
to treat type 2 diabetes and insecticidal agents.
3. Conclusions
The ethanol extract of V. fordiae exhibited insecticidal and α-glucosidase inhibitory activities,
which suggested the potential presence of bioactive secondary metabolites. Bioassay guided
fractionation and purification obtained two new compounds (1, 2) and seven known com-
pounds (3–9). Compounds 3, 4, 6, and 7 were found for the first time in genus Viburnum. In
bioactivity assays, compound 1 showed potent insecticidal activity against M. separata, and
2, 5, 6, 8 and 9 showed potent α-glucosidase inhibitory activity. In conclusion, these results
provided initial evidence of V. fordiae with implication towards potential insecticidal and
α-glucosidase inhibitory properties.

NATURAL PRODUCT RESEARCH
5
Supplementary material
Experimental details, MS, IR and NMR spectra, and data of bioactivity assays are available alongside
Figures S1–S18 as well as Tables S1 and S2, respectively.
Acknowledgements
We are also grateful to the Analytical Detective Center, Yangzhou University, for recording MS, IR and
NMR spectra.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This work was supported by the National Natural Science Foundation of China [grant number 31201563].
ORCID
Jian-Hua Shao
Chun-Chao Zhao
http://orcid.org/0000-0003-2519-6735
http://orcid.org/0000-0001-7941-8175
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