Fitoterapia 72 Ž2001.

676᎐679

Short report

A novel cytotoxic flavonoid glycoside from

Physalis angulata夽
N. IsmailU , M. Alam
College of Pharmacy, Uni¨ ersity of Houston, 4800 Calhoun, Houston, TX 77204-5515, USA

Received 22 January 2001; accepted in revised form 10 February 2001

Abstract

A new flavonol glycoside, myricetin 3-O-neohesperidoside Ž1. was isolated from a cyto-
toxic MeOH extract of the leaves of Physalis angulata. Compound 1 showed remarkable
cytotoxicity in vitro against murine leukemia cell line P-388, epidermoid carcinoma of the
nasopharynx KB-16 cells, and lung adenocarcinoma A-549 with ED50 values of 0.048, 0.50
and 0.55 ␮g mly1, respectively. 䊚 2001 Elsevier Science B.V. All rights reserved.

Keywords: Physalis angulata; Flavonol glycosides; Myricetin 3-O-neohesperidoside; Cytotoxicity

Plant. Physalis angulata L. ŽSolanaceae., leaves collected in June 1997, were

authenticated specimens cultivated and supplied by Moody Gardens, Texas, USA.
The plant commonly called ‘mullaca’ in Peru and was selected for study when an
MeOH extract of the leaves displayed in vitro cytotoxic activity against various
tumor cells.

Uses in traditional medicine and reported activities. Rural inhabitants in the

Peruvian Amazon use the leaf for liver problems, malaria, and hepatitis w1,2x.
Ethanol extracts of the whole plant have shown strong in vitro and in vivo cytotoxic

夽
Dedicated to M. Alam
U
Corresponding author. Present address: Department of Chemistry, University of Saskatchewan, 110
Science Place, Saskatoon, SK S7N-5C9, Canada.
E-mail address: nargisi@hotmail.com ŽN. Ismail..

0367-326Xr01r$ - see front matter 䊚 2001 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X Ž 0 1 . 0 0 2 8 1 - 7
 N. Ismail, M. Alam r Fitoterapia 72 (2001) 676᎐679 677

activity against numerous types of cancer cells including leukemia, lung, colon,
cervix and melanomas w3,4x.

Previously isolated classes of constituents. Steroids w5᎐7x, alkaloids w8x.

New-isolated constituents. Myricetin 3-O-neohesperidoside Ž1. w9᎐13x Žyield:

0.034% on dried wt... A myricetin 3-O-rhamnoglucoside was previously isolated
from Ammi ¨ isnaga w14x, but the authors did not specify the sugar interlinkage.

Myricetin 3-O-neohesperidoside Ž1.. Yellow amorphous powder; mp 174᎐175⬚C;

w ␣ x 25
D -79.4⬚ c 0.7, MeOH ; UV max MeOH : 266 lg␧ 4.23 , 299 sh 3.80 , 356
Ž . Ž . Ž . Ž .
Ž4.12.; Žq NaOMe. 273, 323, 415; ŽqAlCl 3 q HCl. 271, 360 sh, 405 nm; 1 H-NMR
Ž300 MHz, CD3 OD.: ␦ 6.23 Ž1H, d, J 1.8 Hz, H-6., 6.40 Ž1H, d, J 1.8 Hz, H-8., 7.18
Ž2H, s, H-2⬘,6⬘., 5.75 Ž1H, d, J 7.8 Hz, H-1⬙ ., 3.42᎐3.69 Ž5H, m, H-2⬙,6⬙ ., 5.13 Ž1H,
d, J 1.5 Hz, H-1⬘⬙ ., 3.78᎐3.88 Ž4H, m, H-2⬘⬙,5⬘⬙ ., 1.02 Ž3H, d, J 6 Hz, H-6⬘⬙ ., 13.54
Ž1H, s, 5-OH.; 13 C-NMR Ž300 MHz, CD3 OD.: 158.10 ŽC-2., 134.41 ŽC-3., 179.32
ŽC-4., 163.50 ŽC-5., 99.81 ŽC-6., 166.33 ŽC-7., 94.82 ŽC-8., 158.32 ŽC-9., 104.62
ŽC-10., 121.51 ŽC-1⬘., 110.92 ŽC-2⬘ and C-6⬘., 146.50 ŽC-3⬘ and C-5⬘., 138.42 ŽC-4⬘.,
100.67 ŽC-1⬙ ., 80.93 ŽC-2⬙ ., 76.82 ŽC-3⬙ ., 73.52 ŽC-4⬙ ., 77.50 ŽC-5⬙ ., 61.84 ŽC-6⬙ .,
102.53 ŽC-1⬘⬙ ., 75.20 ŽC-2⬘⬙ ., 70.33 ŽC-3⬘⬙ ., 71.92 ŽC-4⬘⬙ ., 69.80 ŽC-5⬘⬙ ., 17.24
ŽC-6⬘⬙ .; yveFABMS mrz: 625wM y Hxy Ž85., 317 wM y 309x Ž100.. Acid hydroly-
sis gave myricetin, glucose and rhamnose.

Tested material. MeOH extract Žyield: 23.7%. and myricetin 3-O-neohesperido-

side.
678 N. Ismail, M. Alam r Fitoterapia 72 (2001) 676᎐679

Table 1
In vitro cytotoxicity of Physalis angulata leaf MeOH extract and myricetin 3-O-neohesperidoside on
P-388, KB-16 and A-549 cells

Tested material ED50 Ž␮grml.a

P-388 KB-16 A-549

MeOH extract 2.50 3.15 3.93

Myricetin 3-O-neohesperidoside 0.048 0.50 0.55
a
ED50 , concentration of the test compound causing 50% reduction of absorbance compared to
untreated cells.

Used microorganisms. The P-388 cells were cultured in Fisher’s medium supple-
mented with 10% heat-inactivated fetal calf serum ŽFCS.. The KB-16 cells were
maintained in the Basal Medium Eagle ŽBME. containing 10% heat-inactivated
FCS. The A-549 cell line was cultured in Eagle’s Minimum ŽEMEM. containing
Earle’s salts and supplemented with 0.1 mM of nonessential amino acids and 10%
heat-inactivated FCS. All the cell lines were maintained in an incubator at 37⬚C in
the humidified air containing 5% CO 2 .

Studied activity. Cytotoxic activity w15x. Test compounds were dispensed to the
established culture plate at eight concentrations each with three replicates. After 3
days of incubation, P-388 cells were evaluated with MTT. KB-16 and A-549 cells
were enumerated using MTT after exposure to test samples for 3 and 6 days,
respectively. Fifty microliters of 1 mgrml MTT were added to each well, and plates
were incubated at 37⬚C for a further 4 h.

Results. Reported in Table 1.

Conclusions. Both the MeOH extract of P. angulata leaves and myricetin 3-O-
neohesperidoside showed strong cytotoxicity against the three tested cell lines, the
strongest action being observed with lymphocytic leukemia P-388 cells. These
results confirm the importance of the investigation on the therapeutic potential of
plants used by indigenous people of Peru.

Acknowledgements

This work was financially supported by the Moody Foundations grant 噛1-5-55279.
The passing in 1999 of the co-author of this paper, Dr Maktoob Alam, is noted
with deep regret.

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