Iraqi J Pharm Sci, Vol.22(2) 2013 Antimicrobial activity of hydroxymatairesinol lignan
30
Antimicrobial Activity of Hydroxymatairesinol (HMR) Lignan
#
Widad M.K. Al-Ani
*,1
and Fitua M. Aziz
*
*
Department of Pharmacognosy and Medicinal Plants, College of Pharmacy, Al-Mustansiriyah University, Baghdad, Iraq.
Abstract
Lignans are natural products widely distributed in the plant kingdom. They are composed of two β-
β-linked phenylpropane (shikimate-derived biogenetic subunits). Although the backbone of lignans is
composed of phenylpropane units, there is enormous diversity in the structure of lignans leading to
different classes of lignans, such as γ-butyrolactone derivatives, eg. Hymatairesinol, bicyclooctadiene
derivatives, e.g. pinoresinol, tetrahydrofuran derivatives e.g.lariciresinol, di-arylbutandiol derivatives,
e.g. secoisolariciresinol. Introduction of a further carbon –carbon linkage leads to a class of lignans
collectively known as cyclolignans such as tetrahydro-naphthalene derivatives, for example
podophyllotoxin. Lignans have a broad range of biological activities; many of them show significant
antitumour, antimitotic, and antiviral effects. They also have cardiovascular effects, antimicrobial and
insecticidal activities. The efflux mechanisms of bacteria to some antibiotics and resistance of bacteria
to many antibiotics led to search for antibacterial compound of plant origin to overcome these
problems. The antibacterial activity of HMR lignan was determined using the disc diffusion method
and the MIC of the isolated compound was tested by the broth micro-dilution method. HMR lignan
showed activity against Staphylococcus epidermidis(17 mm), Candida albicans (13mm),
Proteussp(12mm) and Klebsiella sp (12mm).
Key words: Lignans, Hydroxymatiaresinol, Antimicrobial activity.
هيدروكسي ميتارزينول نكنان كمضاد نهمايكروبات
امم انعانيوداد مصطفى ك
،*1
و فتوة منور عسيس
*
اىعشاق . بغذاد،فشع اىعقاقُش واىْباتاث اىطبُت ، ميُت اىصُذىت ، اىجاٍعت اىَستْصشَت ،
الخالصة
. هْاك phenylpropaneاىينْاُ ٍشمباث طبُعُت راث اّتشاس واسع فٍ اىطبُعت وهٍ تتنىُ فٍ اىْباتاث ٍِ استباط جضئُتُِ ٍِ
بُىىىجُتوغُشها . اىينْاُ ٍشمباث راث فعاىُت HMRتْىع مبُش فٍ تشمُب اىينْاُ عيً اىشغٌ ٍِ اّها تتشابه فٍ هُنيها اىشئُسٍ ٍثو
. اصداد ومزىل ىها فعاىُت فٍ عالج اٍشاض اىقيب واألوعُت اىذٍىَت واىفُشوطيسشطاُ وأخشي ٍضادة ىيبنتشَا ٍتعذدة فبعضها ٍضادة ى
اهتَاً اىباحثُِ فٍ اىسْىاث االخُشة فٍ اىبحث عِ ٍشمباث ٍضادة ىيبنتشَا راث اصو ّباتٍ طبُعٍ ورىل الصدَاد ظاهشة اىَقاوٍت ٍِ
قبو اىبنتشَا ىيَضاداث اىحُىَت ومزىل ظهىس سالالث ٍِ اىبنتشَا ىها قابيُت عيً سحب اىذواء وسٍُه خاسج اىخيُت ٍَا َؤدٌ اىً عذً
ضذ اىبنتشَا بطشَقت االّتشاس. اظهش اىَشمب ّشاط ضذ اىبنتشَا ٍتَثال بىجىد ٍْع َّى HMR. َهتٌ اىبحث بتحذَذ ّشاط اه فعاىُته
اىبنتشَا حىه ّقطت تىاجذ اىَشمب اىْباتٍ.
. مفتاح انكهمات : انهكنان، هيدروكسي ميتاريسينول، مضادات انمايكروبات
Introduction
Lignans are natural products widely
distributed in the plant kingdom. They are
composed of two β-β-linked phenylpropane
(shikimate-derived biogenetic subunits).
Lignans are biosynthesized by the coupling of
two conifer
(l)
alcohol units through a one
electron oxidative coupling process (Figure
1).Lignans have a broad range of biological
activities; many of them show significant
antitumour, antimitotic, and antiviral effects.
They also have cardiovascular effects,
antimicrobial and insecticidal activities
(2
).In
the early 1980s, it was suggested that a diet
rich in HMR lignan may help prevent breast
cancer through conversion to enterolactone
(ENL)
(3)
.An epidemiological study indicated
that ENL
found in the serum of women with no previous
history of breast cancer but not in the serum of
women who have such a history. This study
led to the suggestion that a diet high in lignans
may have chemopreventive effects
(4)
. HMR
lignans usually occur in the form of glycosides
in the fiber of cereals and some fruit like
apricot
(5)
.The knots of the Norway spruce,
from Finland, are considered to be a rich
source of HMR. Concentrations of HMR up to
20% have been found in some knots, making
the Norway sprucea source of HMR for
experimental studies
(6)
.The precipitation of (-)-
HMR by potassium acetate as a potassium a
adduct from a concentrated ethanolic extract,
by Freudenberg, is a superior method for large-
scale production
(7)
.
# Based on oral presentation in the fourteenth professional and scientific conference of the syndicate of
Iraqi pharmacists 19-21 March 2013
1
Corresponding author E-mail:wmkalani@yahoo.com
Received: 4/5/2013
Accepted: 9/6/2013
Iraqi J Pharm Sci, Vol.22(2) 2013 Antimicrobial activity of hydroxymatairesinol lignan
31
Figure(1):Biosynthesis and example of lignans
HMR had stronger lipid peroxidation
inhibition capacity than any other lignan or
flavonoid tested
(8)
.HMR was compared to the
well-known antioxidants TROLOX (a water-
soluble vitamin E derivative), butylated
hydroxyanisole, and butylated hydroxytoluene
in terms of their ability to inhibit lipid
peroxidation, inhibit LDL oxidation, and
scavenge superoxide and peroxyl radicals.
HMR was found to be the strongest
antioxidant, more effective than butylated
hydroxyanisole or butylated hydroxytoluenein
all assays and stronger than TROLOX in all
assays except in the lipid peroxidation
inhibition assay in which the compounds were
almost equally active
(9)
.
The oestrogenic activity of HMR was
investigated by measuring their effects on
growth and apoptotic markers in the human
oestrogen-sensitive cell line MCF-7 in
comparison to oestradiol (E2). HMR and ENL
targeted the same intracellular mechanisms
acted upon by E2 but had a milder effect. This
weak oestrogenic activity was blocked by the
anti-oestrogenic drug tamoxifen
(10)
.It was
found that HMR decreases flushing in women
during menopause, which indicates that this
compound mimics oestrogen activity
(11)
.
Searching for antimicrobial natural products
have been acquired a considerable importance
nowadays due to the development of multiple
drug resistance and efflux mechanisms of
bacteria to many antibiotics
(12)
.Side effects
associated with use of synthetic antibiotics are
frequently more reported than that of natural
products
(13)
.
Three lignans were isolated and
characterized from Larreatridentata and
compounds were tested against 16 bacterial
species/strains. Results showed that:
dihydroguaiaretic (4) acid had activity towards
methicillin resistant (MR) Staphylococcusaur-
eusand multidrug-resistant (MDR) strains of
Mycobacteriumtuberculosis
(14)
.
The relationship between the
stereochemistry and antimicrobial activity of
butane-type lignans was clarified by Satoshi et
al. All stereoisomers of dihydroguaiaretic acid
(4) showed both antibacterial and antifungal
activity. No activity of any stereoisomer of
secoisolariciresinol (5) was apparent
(15)
.
4
5
Lignans secoisolariciresinol 5, pinoresinol 6
and Lariciresinol 7, were isolated from MeOH
extract from Araucaria araucana wood for the
first time in this species and their structures
determined with spectroscopic methods. The
antimicrobial activities of these compounds
were determined for the bacteria Citrobacter
sp., Bacillus subtilis, Escherichia coli,
Micrococcus luteus, Staphylococcus
aureus,and Pseudomonas aeruginosa, and for
the white rooting and staining fungi Mucor
miehei,Paecilomyces variotii, Ceratocystis
pilifera, Trametes versicolor, and Penicillium
notatum, in addition, the MeOH extract was
evaluated against Aspergillus niger, Candida
albicans,Fusarium moniliforme, F.
sporotrichum and Trichophyton
mentagrophytes. The most sensitive bacteria
against pinoresinol were the Gram-positive.
However, secoisolariciresinol exhibited
antifungal activity against wood rooting
(16)
.
Iraqi J Pharm Sci, Vol.22(2) 2013 Antimicrobial activity of hydroxymatairesinol lignan
32
6
7
Materials and Methods
General
HMR potassisum adduct from Linea
R
Company was converted to free HMR lignan
by dissolving in phosphate buffer pH 4.5.
Free HMR from potassium adducts
The food supplement HMR potassium adducts
(1 g) was dissolve in minimum amount
ofaqueous ethanol. Phosphate buffer pH 5(100
mL) was added and the pH was adjusted to 4.5.
The mixture was stirred for 15 minutes and the
free HMR was extracted with ethyl acetate (3 x
100 mL). The combined ethyl acetate layers
were dried with anhydrous magnesium
sulphate, evaporated under vacuo to give free
HMR (7.8 g, 78% yield). The purity of the
lignan was tested by TLC (DCM / ethanol,
7%).
Screening of antibacterial activity
HMR was dissolved in methanol to obtain
final concentrations of (100, 50, and 25) mg /
mL and sterilized by filtration through a 0.45
μm membrane filter. The Muller-Hinton agar
was inoculated with one of the tested bacteria.
Methanol was used as a control to eliminate
solvent effect. Ciprofloxacin (5 μg/ disc) was
used as a positive control.
The agar diffusion method was used to
determine antimicrobial activity of the tested
lignan (HMR).Six mm diameter wells were
punched in to the agar. Inocula were obtained
from overnight cultures on nutrient agar slants
at 37
o
C and diluted in sterile saline solution to
a final concentration of 10
(6)
colony forming
uints (cfu)/ mL (adjusted according to the
turbidity of 0.5 Mc farland scale tube).
Results
In this study HMRlignan showed an
efficient antimicrobial activity against most of
the bacteria and fungi used except
Pseudomonasaeruginosa .This bacteria
showed resistance to all concentration used.
The most sensitive microorganism was
Staphylococcus epidermidis (inhibition zone,
17, 15, 8 mm) and Candida albicans showed
(inhibition zone, 13, 12, 10 mm). Klebsiella sp
and Proteus sp. also showed sensitivity to all
concentrations. Escherichia coli was sensitive
to a high concentration only (100 mg/ mL).
The result of our study revealed that HMR
lignan posses antimicrobial properties as
antibiotic principles (Table 1)
Table (1): Inhibition zones of bacterial growth produced by HMR lignan
Methanol as a solvent showed no antimicrobial
activity against the bacteria, while HMR lignan
showed a good inhibition zone for both
(Staphylococcus epidermidis) and Klebsiella sp
(Figure 2).
Microorganisms
Concentrations (mg/mL)/ Inhibition zone in mm
100
mg/mL
50
mg/ mL
25
mg/ mL
Solvent (C) ciprofloxacin
Proteus sp. 12 9 7 - 35
Pseudomonasaeruginosa - - - - 27
Escherichia coli 8 - - - 32
Staphylococcus aureus 8 7 - - 19
Candida albicans 13 12 10 - -
Klebsiella sp 12 9 7 - -
Staphylococcus
epidermidis
17 15 8 - -
Iraqi J Pharm Sci, Vol.22(2) 2013 Antimicrobial activity of hydroxymatairesinol lignan
33
Figure (2): Inhibition zone for staphylococcus
epidermidis
Discussion
Natural products have been found to be an
efficient antimicrobial compounds especially
those with phenolic group. It is necessary to
investigate the activity of the diphenolic HMR
lignan to improve the quality of health care.
Many organisms develop resistance to
antibiotics which lead to search for novel
antimicrobial agents. References 14 and 15
focus on the antimicrobial activity of butane
type lignan therefor HMR is also a good
example for those butane type antimicrobial
lignans.
The diameter of inhibition zone of the
antibacterial agents the lignans and
ciprofloxacin were different according to the
kinds, concentrations and purity.
The mechanism of action of the antimicrobial
(HMR) is due to the toxic effect or may be
impair variety of enzyme systems including
those involved in energy production and
structural component synthesis
(17)
.
Excess acid should be avoided during the
libration of free HMR from the potassium
adduct due to cyclization of HMR to the
cyclolignan (Figure 3)
(18)
.
Figure (3): Cyclization of HMR to cyclolig-
nansin acidic media
Conclusion
HMR lignan shoswed an efficient
antimirobial activity it shows antimicrobial
activity against most of the bacteria and fungi
used. This result together with the results
reported in the literatures support the idea of
using natural compounds as sources for
antimicrobial drug to overcome the resistance
and efflux mechanism of microbe to
antibiotics.
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