Kutaiba Ibrahim Alzand et al.

/ Journal of Pharmacy Research 2012,5(8),4013-4020

Review Article Available online through
ISSN: 0974-6943 http://jprsolutions.info
Flavonoids: Chemistry, Biochemistry and Antioxidant activity
Kutaiba Ibrahim Alzand a* and Mohamed Abdalkarim Mohamedb *
a
Department of Drug Technology, Faculty of Medical Technology, Derna, Libya.
b
Department of Chemistry, Sudan University of Science and Technology, Khartoum, Sudan.
Received on:09-05-2012; Revised on: 14-06-2012; Accepted on:22-07-2012

ABSTRACT
Flavonoids continue to capture the interest of scientists from many different disciplines because of their structural diversity, biological and ecological
significance (e.g. as the coloured pigments in many flower petals), and health-promoting and anti-cancer properties. According to our assessment, approxi-
mately 9000 different flavonoids have already been reported from plant sources, but no doubt many more remain to be discovered. The topics that will be
discussed in this review are chemistry, biosynthesis, bioavailability and metabolism, antioxidant and estrogenic properties of flavonoids.

Key words: Flavonoids, Chemistry of Flavonoids, Antioxidant activity of Flavonoids

1. INTRODUCTION

1.1. PRIMARY AND SECONDARY METABOLITES

Plant chemicals are often classified as either primary or secondary metabo-
lites. Primary metabolites are substances widely distributed in nature, occur-
ring in one form or another in virtually all organisms. In higher plants such
compounds are often concentrated in seeds and vegetative storage organs and
are needed for physiological development because of their role in basic cell
metabolism. (Examples: carbohydrates, lipids, proteins…)1.
Secondary metabolites are compounds biosynthetically derived from pri-
mary metabolites. Contrary to primary metabolites these compounds are
not ubiquitous in the living organisms that produce them nor are they neces-
sarily expressed continuously. Although plants are better known as a source
of secondary metabolites, bacteria, fungi and many marine organisms (sponges,
tunicates, corals, and snails) are very interesting sources, too. Secondary
metabolites, also known as natural products are not essential for normal
growth, development or reproduction of an organism. In this sense they are
“secondary”. The function or importance of these compounds to the
organism’s development is usually of ecological nature as they are used as
defence against predators (herbivores, pathogens etc.), for interspecies com-
petition, and to facilitate the reproductive processes1,2.
Secondary metabolites can be classified by their chemical structure or physi-
cal properties into one or more of the following groups: alkaloids, phenols,
terpenoids, iridoids, steroids, saponins, polyletides, aliphatic, aromatic, and
heteroaromatic organic acids, peptides, ethereals oils, resins and balsams 2.
Phenolic compounds form one of the main classes of secondary metabolites
with a large range of structures and functions, but generally possessing an
aromatic ring bearing one or more hydroxyl substituents. This definition is
not entirely satisfactory, however, since it inevitably includes compounds
such as oestrone, the female sex hormone that is principally terpenoid in
origin. For this reason, a definition based on metabolic origin is preferable,
the plant phenols being regarded as those substances derived from the
shikimate pathway and phenylpropanoid metabolism3.
*Corresponding author.
Kutaiba Ibrahim Alzand
Department of Drug Technology,
Faculty of Medical Technology,
Derna, Libya
Natural polyphenols can range from simple molecules, such as phenolic
acids, to highly polymerized compounds, such as tannins. They occur pri-
marily in conjugated form, with one or more sugar residues linked to hy-
droxyl groups, although direct linkages of the sugar unit to an aromatic carbon
atom also exist. The associated sugars can be present as monosaccharides,
disaccharides, or even as oligosaccharides, and glucose is the most common
sugar residue. Associations with other compounds, such as carboxylic and
organic acids, amines, and lipids, and linkage with other phenols are also
common4.
Polyphenols can be divided into at least 10 different classes depending on
their basic chemical structure5. Table 1 illustrates the basic chemical struc-
ture of the main polyphenolic compounds. In order to simplify the drawing
of chemical structures, only the skeleton structures are shown in Tables 1,
and there should be at least one hydroxyl group attached to the aromatic ring.
1.2. FLAVONOIDS
1.2.1. CLASSIFICATION AND CHEMISTRY
Flavonoids are phenolic substances isolated from a wide range of vascular
plants, and more than 8150 different flavonoids have been reported5. Fla-
vonoids are located inside the cells or on the surface of various plant organs
and have various functions in plants6. They act in plants as antioxidants,
antimicrobials, photoreceptors, visual attractors, feeding repellents, and for
light screening7. Many studies have shown that flavonoids exhibit biological
and pharmacological activities, including antioxidant, cytotoxic, anticancer,
antiviral, antibacterial, cardioprotective, hepatoprotective, neuroprotective,
antimalarial, antileishmanial, antitrypanosomal and antiamebial properties8-
12
. These biological and pharmacological properties are usually attributed to
their free radical scavenging efficacies, metal complexion capabilities, and
their ability to bind to proteins with a high degree of specificity13.
The basic flavonoid structure contains the flavan nucleus (1), which consists
of 15 carbon atoms derived from a C6-C3-C6 skeleton. A flavonoid skeleton is
composed of two aromatic rings (commonly designated as A and B), which
are linked by a three-carbon chain. The connecting carbon chain combines
with an oxygen to form a heterocyclic central or C-ring for most flavonoids
with the exception of chalcones (2) in which the carbon chain between the A

Journal of Pharmacy Research Vol.5 Issue 8.August 2012 4013-4020

 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
and B rings is linear14. The numbering scheme for chalcones differs from
three-ring flavonoids in that the A ring, rather than the B ring carbons are
labeled as prime.
Depending on the position of the linkage of the aromatic B-ring to the
benzopyrano (chromano) moiety, this group of natural products may be
divided into three classes: the flavonoids (2-phenylbenzopyrans) 1,
isoflavonoids (3-phynylbenzopyrans) 3, and the neoflavonoids (4-
phynylbenzopyrans) 4. These groups usually share a common chalcone
precursor, and therefore are biogenetically and structurally related2.
Based on the degree of oxidation and saturation present in the heterocyclic C-
ring, the flavonoids may be divided into the following groups: flavone (5),
flavonol (6), flavanone (7), dihydroflavonol (8), flavan (1), flavan-3-ol (9),
flavan-4-ol (10), flavan-3,4-diol (11), and anthocyanidin (12).
5 6 7 defense against microorganisms and germination of pollen20,21.
8 9
10 11 12
Natural products such as chalcones (2), dihydrochalcones (13), aurones (14)
and auronols (15) also contain a C6-C3-C6 backbone and are considered to be
minor flavonoids6. Natural products such as chalcones (2), dihydrochalcones
(13), aurones (14) and auronols (15) also contain a C6-C3-C6 backbone and
are considered to be minor flavonoids6.
13 14 15
Journal of Pharmacy Research Vol.5 Issue 8.August 2012
The flavonoids may be modified by hydroxylation, methoxylation, or O-
glycosylation of hydroxyl groups as well as C-glycosylation directly to
carbon atom of the flavonoid skeleton. In addition, alkyl groups (often prenyls)
may be covalently attached to the flavonoid moieties, and sometimes addi-
tional rings are condensed to the basic skeleton of the flavonoid core. Fla-
vonoid glycosides are frequently acylated with aliphatic or aromatic acid
molecules. These derivatives are thermally labile and their isolation and fur-
ther purification without partial degradation is difficult. Condensed tannins
create a special group of flavonoid compounds formed by polymeric com-
pounds built of flavan-3-ol units, and their molecular weights often exceed-
ing 1,000 Da11.
Up until the middle of the 20th century, flavonoids were believed to be waste
products of plant primary metabolism, a notion that was soon abandoned
based on research demonstrating the myriad functions of flavonoids in plant
survival16. We now know the complex metabolic pathways used to synthe-
size flavonoids in plants-pathways that have evolved over millennia to pro-
vide a survival advantage to plants. Plants, clearly do not have the luxury of
mobility and have had to evolve elaborate chemical mechanisms in order to
defend themselves from various insults (such as UV radiation) and in order to
attract those that will assist with their reproduction (bees, birds)17. Indeed,
the complexity of the plant genome, which exceeds that of humans, is be-
lieved to have developed to maintain these immensely complex pathways of
synthesis of novel chemicals17.
Flavonoids are best known as the red, blue and purple pigments of flowering
plants (due to the anthocyanidin sub-group) although the red pigment of
some fruits can be due to carotenoids such as lycopene18. These pigments
and the yellow pigments of flavones and flavanols are also responsible for
the fall leaves coloration19. Because of the importance of color in pollinator
attraction, flavonoids have an important role in plant reproduction. Fla-
vonoids in plants also serve to protect from ultraviolet (UV) light owing to
their high UV absorbance coefficients, and it has been suggested that this
property of flavonoids was critical in the evolution of aquatic plants to a
terrestrial existence16. Flavonoids also possess other critical functions in
Each group of flavonoids possesses unique chemical properties and has a
particular distribution in plants. Anthocyanins (glycosylated anthocyanidins)
and proanthocyanidins (polymers that produce anthocyanidins when hy-
drolyzed) primarily provide color to flowering plants and fruit, and are
therefore found in high concentrations in the skin of red grapes, red wine and
berries. Flavan-3-ols, such as catechin, epicatechin gallate are colorless and
are found in high concentrations in green tea. Isoflavones are only found in
legumes (e.g. soy) and are therefore consumed in high quantities in regions of
the world with high soy consumption. Flavanones are found in high levels in
citrus fruits, while flavones are present in green leafy spices such as parsley,
and flavonols are ubiquitous and found in most fruits and vegetables con-
sumed in the human diet14,22.
Interest in the health benefits of flavonoids stemmed from early research in
1936 by the Hungarian scientist Szent-Gyorgyi, who also incidentally dis-
covered vitamin C. He isolated a substance, which he called citrin, from
lemons that restored weakened capillaries to normal. This effect was not
noted when vitamin C alone was administered. Citrin was later shown to be
composed of the flavonoids hesperidin and eriodictyol23. Although citrin
was also called vitamin P, this name was dropped in the 1950s after it was
concluded that flavonoids did not fit the strict definition of a vitamin24.
Despite not qualifying as vitamins, flavonoids have been shown in the last 72
years since Szent-Gyorgyi’s initial discovery to affect various aspects of
human health not limited to their beneficial effects on capillary wall integrity.
4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
1.2.2. BIOSYNTHESIS OF FLAVONOIDS
Biosynthesis of flavonoids in plants is via a series of enzymatic steps start-
ing with the aromatic amino acid phenylalanine and acetate19. The initial step
in biosynthesis of all flavonoids is the condensation of 4-coumarate coen-
zyme A (shikimate derived, B ring) with three malonyl coenzyme A mol-
ecules (polyketid origin, A ring) to give 2', 4', 6', 4-tetrahydroxychalcone,
which is catalysed by the enzyme chalcone synthase25. The chalcone is then
isomerised to the flavanone naringenin, a key indermediate, which can be
converted to several end-products including aurones, isoflavonoids, flavones,
flavonols, flavandiols, anthocyanins, condensed tannins, and phlobaphene
pigments (Figure 1)25,26.
Table 1. Main Classes of Phenolic Compounds
1.2.3. BIOAVAILABILITY AND METABOLISM OF FLAVONOIDS
The absorption, metabolism and excretion of flavonoids is a complex pro-
cess involving various structural modifications to the ingested flavonoid in
multiple tissues and cellular compartments. Determining the bioavailability
of flavonoids (the proportion of flavonoid found in blood or target tissue
after ingestion) is critical to understanding the effects of flavonoids as
chemopreventive agents. Although flavonoids have been shown to undergo
extensive metabolism, this does not necessarily equate to biological inacti-
vation of the compound. In many cases in pharmacology, the metabolized
product is often more active than the parent compound. A case in point is
morphine whose glucuronated metabolite is known to be a more potent
Journal of Pharmacy Research Vol.5 Issue 8.August 2012
opiate than the parent compound27.
Flavonoids are predominantly absorbed in the small intestine, with only
small amounts absorbed via the gastric mucosa28. The glycosylation state
of the flavonoid greatly affects the mechanism of flavonoid absorption.
Most flavonoids in nature exist as glycosides. Early studies suggested that
flavonoid glycosides were not absorbed intact in the human gut (due to their
high hydrophilicity). These findings have more recently been refuted 29.
Studies have since demonstrated that quercetin glycosides are not only
absorbed, but that their absorption is actually enhanced compared to quer-
cetin aglycone30. This absorption is believed to occur partly due to the
action of sodium dependent glucose transporter (SGLT1)31. More com-
monly, however, the first step in absorption of flavonoid-glycosides is
usually hydrolysis of the sugar moiety in the gut resulting in generation of
the flavonoid aglycone32. This hydrolysis was initially assumed only to
occur in the colon by bacteria since humans lack the necessary enzymes to
hydrolyze the ß-glycoside linkages of flavonoid glycosides. However, re-
cently is has become clear that a broad-specificity ß-glucosidase enzyme in
enterocytes and lactase phloridzin hydrolase in the small intestine brush
border can hydrolyze these ß-glycoside linkages33. Hydrolysis of fla-
vonoid-glycosides has also been shown to occur in the oral cavity34. The
flavonoid aglycones generated by hydrolysis of the sugar moiety are more
lipophilic, and hence are more readily absorbed in the gut by passive diffu-
sion. Flavonoids entering the colon undergo a similar hydrolysis by bacte-
rial glucosidases29. Flavonoids that reach the colon undergo ring scission of
the aromatic ring after sugar hydrolysis, resulting in simple phenolic com-
pounds, which accounts for the low levels of flavonoid absorption from the
colon35.
The type of sugar moiety is an important determinant of absorption effi-
ciency, with flavonoid-glucosides much more readily absorbed than fla-
vonoid-rutinosides36. With respect to the flavonoid aglycones, methoxylated
flavonoids as opposed to hydroxylated flavonoids are much more readily
flavonoid absorption include the protein content of food ingested with the
flavonoid. Since flavonoids bind to proteins, flavonoid absorption will be
attenuated until the protein is digested 38.
The biotransformation of flavonoids continues in the enterocytes. The main
metabolic transformations include conjugation of glucuronic acid
(glucuronidation), methylation and sulphation39. These conjugations are es-
sentially phase II detoxification reactions resulting in increased molecular
mass and improved solubility of the compound which enhances excretion of
the compound in bile and urine40. Thus the enterocyte is an important site of
flavonoid metabolism. Flavonoid aglycones that reach the circulation are
bound to albumin. Interestingly, binding to albumin does not affect the anti-
oxidant ability of flavonoids, an important point in terms of the likely
biological effect of absorbed flavonoids41. Flavonoids entering the circula-
tion subsequently undergo phase II detoxification in the liver. Other transfor-
mations in the liver include the formation of flavonoid-glutathione adducts
also resulting in enhanced excretion of the flavonoid39. Excretion of fla-
vonoids occurs in urine as well as bile42.
The kinetics of flavonoid absorption and metabolism in humans has been
studied for quercetin and other flavonoids. Hollman et al. demonstrated that
peak plasma levels of quercetin were reached in 2.9 hours in subjects con-
suming a meal of 333 grams fried onion43. The mean peak plasma level of
quercetin was 196 ng/ml and the half-life of quercetin was 16.8 hours. This
long half-life suggests that quercetin may accumulate with continued fla-
vonoid administration. Importantly for purposes of chemoprevention, stud-
ies have also demonstrated the accumulation of flavonoids in various animal
tissues44.
4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020

Figure 1. Schematic of the major branch pathways of flavonoid biosynthesis, starting with general phenylpropanoid metabolism and leading to the nine major
subgroups: the colorless chalcones, aurones, isoflavonoids, flavones, flavonols, and flavandiols (gray boxes), and the anthocyanins, condensed tannins, and
phlobaphene pigments (colored boxes). The first committed step is catalyzed by chalcone synthase (CHS), which uses malonyl CoA and 4-coumaroyl CoA as
substrates. Enzyme names are abbreviated as follows: cinnamate-4-hydroxylase (C4H), chalcone isomerase (CHI), chalcone reductase (CHR), chalcone synthase
(CHS), 4-coumaroyl:CoA-ligase (4CL), dihydroflavonol 4-reductase (DFR), 7,2'-dihydroxy, 4'-methoxyisoflavanol dehydratase (DMID), flavanone 3-hydroxy-
lase (F3H), flavone synthase (FSI and FSII), flavonoid 3' hydroxylase (F3’H) or flavonoid 3’5' hydroxylase (F3’5’H), isoflavone O-methyltransferase (IOMT),
isoflavone reductase (IFR), isoflavone 2'-hydroxylase (I2’H), isoflavone synthase (IFS), leucoanthocyanidin dioxygenase (LDOX), leucoanthocyanidin reductase
(LCR), O-methyltransferase (OMT), Phe ammonia-lyase (PAL), rhamnosyl transferase (RT), stilbene synthase (STS), UDPG-flavonoid glucosyl transferase
(UFGT), and vestitone reductase (VR).
Journal of Pharmacy Research Vol.5 Issue 8.August 2012 4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
Quercetin, a flavonol, is believed to have different absorption and kinet-
ics to other flavonoid classes. Anthocyanins, in comparison, are absorbed
poorly and rapidly excreted in urine. Citrus flavanones are well absorbed but
have shorter plasma half lives. They also reach higher maximum concentra-
tions than flavonols45. A significant degree of variability is therefore expected
in the pharmacokinetic properties of different flavonoids. As with other
ingested compounds, inter-individual variability is another important factor
accounting for the pharmacokinetic properties of flavonoids in humans.
Food preparation has variable effects on the bioavailability of flavonoids
in the diet. Peeling, for example, greatly reduces flavonoid content since the
peel contains a large proportion of flavonoids in fruits and vegetables46. The
effect of cooking on flavonoid content has been examined for quercetin con-
tent in onions after cooking. When onions are boiled, flavonoids diffuse out
to enter the broth, making the broth a rich source of flavonoids. Frying
onions for 40 minutes did not alter total quercetin content. An increase in
quercetin is noted on microwaving due to increased extractability47. Con-
sumption of flavonoids and protein together, while postulated to reduce the
absorption of flavonoids, has not been shown to have an effect48. Thus, in
general, flavonoid availability from food appears to be enhanced by cooking.
1.2.4. THE MECHANISMS OF ACTION OF FLAVONOIDS
1.2.4.1. Antioxidant Potential
The ability of flavonoids to function as antioxidants has given them an
important place in the field of human health and medicine. Diets high in
flavonoids, fruits, and vegetables are protective against a variety of diseases,
particularly cardiovascular disease and some types of cancer49. The protec-
tive effects of flavonoids in biological systems are ascribed to their capacity
to transfer free radical electrons, chelate metal catalysts50, activate antioxi-
dant enzymes51, reduce alpha-tocopherol radicals52, and inhibit oxidases53.
The flavones and catechins seem to be the most powerful flavonoids for
protecting the body against reactive oxygen species54. Reactive oxygen spe-
cies (ROS) are highly reactive molecules with both physiologic and patho-
logic roles. ROS can occur in the form of molecules with highly reactive
unpaired electrons known as free radicals (e.g. superoxide, O2.-), or as non-
radicals that are highly liable to form free radicals (e.g. hydrogen peroxide,
H2O2). They can exist in the body as a result of deliberate synthesis (e.g.
production by macrophages for bacterial killing), or as a result of accidental
production by metabolic processes such cellular respiration in mitochondria,
or via exogenous insults such as smoking55. Many ROS are not in themselves
exceptionally reactive; however, in the presence of free heavy metal ions
such as copper and iron they generate highly toxic radicals such as hydroxyl
ions (OH.). ROS are highly damaging as they can attack lipids in cell mem-
branes, proteins, carbohydrates, and DNA. The resulting oxidative damage
may play a role in aging and chronic and degenerative diseases including
cancer56-59.
About 1-3% of the oxygen we breathe ultimately goes into making ROS,
resulting in a huge burden of pro-oxidant free radicals that has to be effec-
tively removed60. The human body relies on both endogenous and exogenous
(dietary) anti-oxidant systems to buffer the effect of the ROS constantly
produced by metabolic processes. Endogenous antioxidant systems include
enzymes such as superoxide dismutase, which converts O2.- into H2O2, and
glutathione peroxidase and catalase, that serve to remove H2O2. Non-enzy-
matic endogenous defense mechanisms have a significant anti-oxidant im-
pact, including buffering by plasma urate and plasma protein thiols. Further-
more, the sequestration of heavy metal ions in binding proteins such as
transferrin (iron) reduces the risk of formation of toxic hydroxyl radicals61
physiology61,62.
Journal of Pharmacy Research Vol.5 Issue 8.August 2012
Antioxidant phytochemicals constitute some of the most important exog-
enous defense antioxidants in mammalian physiology. Up until the mid-
1990’s the dietary phytochemicals most prominently studied for their anti-
oxidant properties were vitamin C, E and the carotenoids63. Polyphenols,
which constitute a major group of plant chemicals, only gained interest for
their antioxidant effects in the last decade. Flavonoids, the largest group of
polyphenols found in nature, appear to be particularly potent antioxidants in
vitro. The flavonoids, however, are not all equally effective, with definite
structural requirements necessary for the greatest antioxidant effect. The
presence of a 2,3 double bond in the C-ring, a catechol structure in the B-ring,
and hydroxylation at position 3 and 5 of the A ring appear to impart increased
redox potential64. The redox potential of quercetin was similar to ascorbic
acid and greater than the redox potential of uric acid65.
Direct scavenging of free radicals is one of the major mechanisms of antioxi-
dant activity by flavonoids. The resulting aroxyl radical (Flavonoid-O.) is
more stable than other ROS and gains further stability on reacting with a
second radical to form a stable quinine structure66. Several other mechanisms
of antioxidant activity of flavonoids have been proposed including scaveng-
ing of transition metal ions67, and inhibition of enzymes responsible for
antioxidant production. In terms of the latter property, flavonoids have been
shown to inhibit several pro-oxidant enzymes including xanthine oxidase68,
glutathione S-transferase69, nitric oxide synthase70, and NADH oxidase71
among others.
1.2.4.2. Pro-Oxidant Effects
While flavonoids are best known for their anti-oxidant properties, it has been
shown that under certain conditions, flavonoids may be pro-oxidant. This
property has been proposed to account for several biological effects of fla-
vonoids, such as apoptosis, that are induced in the setting of oxidative stress.
These pro-oxidant effects have also been observed for other phenolic antioxi-
dants including tocopherols, ascorbate, urate, curcumin and N-acetylcysteine.
The balance between anti-oxidant and pro-oxidant effects of these compounds
is dependent on several factors in the cellular environment, particularly on
the presence of transition metal ions.
The hydroxyl groups of flavonoids account for much of their antioxidant
effect. After scavenging ROS, flavonoids are themselves oxidized, with a
hydroxyl group now containing a free radical known as a phenoxyl radical.
Some flavonoids possess a catechol structure in the B-ring. Oxidation of
these flavonoids can result in a semi-quinone radical. The flavonoid semi-
quinone can undergo further oxidation resulting in flavonoid quinone. There-
fore different types of oxidation of flavonoids occur depending on their exact
structure. In addition to scavenging of ROS, flavonoids can also be oxidized
in other ways. These include oxidation by cellular peroxidases, or by auto-
oxidation in the presence of oxygen- a process greatly accelerated in the
presence of transition metal ions.
The paradox of ‘antioxidant’ flavonoids is that in the process of scavenging
ROS, they become pro-oxidant radicals themselves, albeit less reactive than
the scavenged species. The flavonoid radicals never-the-less have undesirable
properties. For example, in the presence of transition metals such as Cu+2,
flavonoids undergo a series of redox reactions culminating in the genesis of
damaging hydroxyl radicals72. Another mechanism involves the flavonoid-
quinones that are the products of oxidation in catechol containing flavonoids.
These are highly reactive to thiol groups, and result in the formation of
flavonoid conjugates with thiol containing proteins such as glutathione. In-
terestingly, flavonoids that do not form flavonoid-quinones have also been
shown to form thiol conjugates, highlighting the complexity of flavonoid
chemistry that remains to be fully elucidated73. The pro-oxidant effects of
flavonoids have been shown to result in DNA damage and lipid peroxidation
4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
in vitro. Pro-oxidant radicals have been demonstrated for several flavonoids
including myricetin74, quercetin75,76, proanthocyanidins77, green tea cat-
echins78,79, daidzein80 and baicalin81.
1.2.4.2. Hormonal Properties
Together with the antioxidant effects of flavonoids, the hormonal, and par-
ticularly the estrogenic effects of flavonoids have garnered the greatest atten-
tion in flavonoid research over the past 50 years. The estrogenic properties
of flavonoids first came to light in the 1950’s when it was observed that
sheep grazing on red clover pastures had reduced breeding rates82. Red clover
was found to contain several isoflavones and the estrogen-like properties of
isoflavones were shown to account for fertility disturbances in animals feed-
ing on red clover. As a result of their estrogen-like properties, isoflavones are
also classed as phytoestrogens. Flavonoids from the flavone, flavonols,
flavonone and chalcone classes are considerably weaker phytoestrogens than
the isoflavones as determined by competitive binding assays83. The precise
physiologic effects of flavonoids mediated by their binding to ERa and ERß
are yet to be fully determined. This is an important area of future research
because phytoestrogenic flavonoids are consumed in large amounts in the
human diet. If the estrogenic effects of these compounds are predominantly
pro-proliferative at low concentrations, flavonoids could potentially pose a
health risk in terms of promoting hormone dependent cancers.
Flavonoids may also exert anti-estrogenic effects by various enzymatic
mechanisms. Blocking the synthesis of estrogens by inhibiting aromatase is
an established strategy in the treatment of breast cancer. Flavonoids have
been shown to bind the active site of aromatase, and inhibit its function, with
flavones and flavanones, rather than isoflavones having the greatest effect84,85.
Other enzymes of note in estrogen metabolism include sulfatase and 17ß
hydroxydteroid dehydrogenase, both of which result in activation on estra-
diol precursors, and which are inhibited by flavonoids86. The levels of sex
hormone binding globulin (SHBG) by flavonoids is also of importance, as
excess SHBG can bind estrogen reducing its effect87.
The estrogenic effects of isoflavones in humans is supported by studies
demonstrating altered menstrual cycle length in women consuming daily soy
protein, a product rich in isoflavones88-90.
The similarity in structure of flavonoids to all steroid hormones raises the
possibility of ligand binding of flavonoids to other members of the nuclear
steroid receptor family. Flavonoids have been shown to bind and activate a
number of nuclear receptors including androgen91,92, progesterone91, thy-
roid93, and peroxisome proliferator-activated receptor PPAR?93. Flavonoids
functionally activate androgen receptor mediated transcription, resulting in
increased PSA, a major downstream androgen receptor regulated gene. Api-
genin, a flavone, was the most effective flavonoid in up-regulating PSA ex-
pression94. Interestingly, in a related study other flavonoids were shown to
have precisely the opposite effect and inhibit PSA production. It was con-
cluded that unlike the estrogenic effects of flavonoids, the effects on PSA
production did not follow a structure-function relationship 95 .
In addition to their sex steroid effects, flavonoids affect several other hor-
monal pathways. For example, several mechanisms have been described for
the potentiation of the vitamin D pathway by genistein. This includes up-
regulation of vitamin D receptor gene expression and activity96, and inhibi-
tion of enzymes (CYP24) that convert 1,25-vitamin D into less active me-
tabolites 97 . The vitamin D pathway is increasingly implicated in
chemoprevention of prostate cancer, and up-regulation of this pathway is a
potentially useful synergistic property of flavonoids. Genistein has also
been shown to have a stimulatory effect on insulin secretion in vitro (al-
though this effect is not seen in vivo) and an inhibitory effect on leptin
Journal of Pharmacy Research Vol.5 Issue 8.August 2012
secretion in rats administered genistein98. Flavonoids also inhibit corticoster-
oid secretion in vitro and in vivo98. Finally, the goitrogenic activity of soy is
well documented, especially in the setting of iodine deficiency. This effect is
believed to be secondary to inhibition of thyroid peroxidase, a major metabo-
lizing enzyme in thyroxine biosynthesis99. Overall, the interactions of fla-
vonoids with steroid hormone pathways are highly complex. However, the
effect of flavonoids on these pathways has been shown to ultimately cause
alterations resulting in beneficial effects such as the negative regulation of
proliferative stimuli. As with much of flavonoid research, the in vivo effects
of flavonoids on hormonal signaling require further study. This is highlighted
in a recent review by Hamilton-Reeves et al, where the majority of interven-
tion studies reviewed did not find a difference in circulating sex steroid
hormone levels100.
REFERENCES
1. Balandrin, F.M., Klocke, J.A., Wurtele, E.S., and Bollinger, W.H.,
Sciences, 228, 1154 (1995).
2. Dewick, P.M., Medicinal Natural Products. A Biosynthetic Ap-
proach, 2 nd ed., John Wiley & Sons, West Sussex 2001.
3. Robard, K., Prenzler, P. D., Tucker, G., and Glover, W., Food
Chemistry, 66, 401 (1999).
4. Bravo, L., Nutrition reviews, 56, 317 (1998).
5. Harborne, J.B., General procedures and measurement of total
phenolics, in Methods in Plant Biochemistry, Vol. 1, Plant Pheno-
lics, Harborne, J.B., Ed., Academic Press, London, 1989, chap. 1.
6. Marais, J.P.J., Deavours, B., Dixon, R.A., and Ferreira, D., The
stereochemistry of flavonoids, in The Science of Flavonoids,
Grotewold, E., Ed., Springer Science, New York, 2006, chap. 1.
7. Pieatta, G.P., Flavonoids as antioxidant, J. Nat. Prod., 63, 1035
(2000).
8. Harborne, J.B., and Williams, C.A., Phytochemistry, 55, 481
(2000).
9. Nowakowska, Z., Eur. J. Med. Chem., 42, 125 (2007).
10. Williams, C.A., Harborne, J.B., Geiger, H., and Hoult, J.R.S.,
Phytochemistry, 51, 417 (1999).
11. Weimann, C., Goransson, U., Ponprayoon-Claeson, P., Bhlin, L.,
Rimpler, H., and Heinrich, M., J. Pharm. Pharmacol., 54, 99
(2002).
12. Havsteen, B.H., Pharmacol. Therapeutics, 96, 67 (2000).
13. Snow, R.W., Guerra, C.A., Noor, A.M., Myint, H.Y., and Hay,
S.I., Nature, 434, 214 (2005).
14. Beecher, G.R., J. Nutr., 133, 3248S (2003).
15. Stobiecki, M., Kachlicki, P., Isolation and identification of fla-
vonoids, in The Science of Flavonoids, Grotewold, E., Ed., Springer
Science, New York, 2006, chap. 2.
16. Andersen, Ø.M., and Markham, K.R., Flavonoids:
Chemistry,Biochemistry and Applications, CRC Press: Boca
Raton, 2006.
17. Prasain, J.K., and Barnes, S., Mol. Pharm., 4, 846 (2007).
18. Timberlake, C.F., and Henry, B.S., Endeavour, 10, 31 (1986).
19. Winkel-Shirley, B., Plant physiology 126, 485 (2001).
20. Taylor, L.P., and Grotewold, E., Current opinion in plant biology,
8, 317 (2005).
21. Treutter, D., Plant biology, 7, 581 (2005).
22. Harnly, J.M., Doherty, R.F., and Beecher, G.R., Journal of agri-
cultural and food chemistry, 54, 9966(2006).
23. Bentsath, A., St.Rusznyak, I., and Szent-Gyorgyi, A., Nature,
138 (1936).
24. Harborne, J.B., The Flavonoids: Advances in Research Since
1986, Chapman and Hall, London, 1994.
25. Strack, D., and Wray, V., The Anthocyanins, in The Flavonoids:
4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
Advances in Research since 1986, Harborne, J.B., Ed., Chapman
and Hall, London, 1994, chap.1.
26. Cooper-Driver, G.A., Phytochemistry, 56, 229 (2001).
27. Wittwer, E., Kern, S.E., The AAPS journal, 8, 348 (2006).
28. Piskula, M.K., Yamakoshi, J., and Iwai, Y., FEBS letters, 447, 287
(1999).
29. Griffiths, L., and Barrow, A., The Biochemical Journal, 130, 1161
(1972).
30. Hollman, P.C., de Vries, J.H., van Leeuwen, S.D., The American
journal of clinical nutrition, 62, 1276 (1995).
31. Walgren, R.A., Lin, J.T., Kinne, R.K., and Walle, T., The Journal
of pharmacology and experimental therapeutics, 294, 837 (2000).
32. Spencer, J.P., Chowrimootoo, G., Choudhury, R., Debnam, E.S.,
Srai, S.K., and Rice-Evans, C., FEBS letters, 458, 224 (1999).
33. Day, A.J., Gee, J.M., DuPont, M.S., Johnson, I.T., and Williamson,
G., Biochemical pharmacology, 65, 1199 (2003).
34. Walle, T., Browning, A.M., Steed, L.L., Reed, S.G., and Walle,
U.K., J. Nutr, 135, 48 (2005).
35. Rice-Evans, C., Current medicinal chemistry, 8, 797 (2001).
36. Hollman, P.C., Bijsman, M.N., van Gameren, Y., Cnossen, E.P.,
de Vries, J.H., and Katan, M.B., Free radical research, 31, 569
(1999).
37. Walle, T., Seminars in cancer biology, 17, 354 (2007).
38. Prasain, J.K., and Barnes, S., Mol Pharm, 4, 846 (2007).
39. Spencer, J.P., Abd-el-Mohsen, M.M., and Rice-Evans, C., Ar-
chives of biochemistry and biophysics, 423, 148 (2004).
40. Williamson, G., Day, A.J., Plumb, G.W., and Couteau, D., Bio-
chemical Society transactions, 28, 16 (2000).
41. Janisch, K.M., Williamson, G., Needs, P., and Plumb, G.W., Free
radical research, 38, 877 (2004).
42. Nielsen, I.L., and Williamson, G., Nutr Cancer, 57, 1 (2007).
43. Hollman, P.C., vd Gaag, M., Mengelers, M.J., van Trijp, J.M., de
Vries, J.H., and Katan, M.B., 21, 703 (1996).
44. Chu, K.O., Wang, C.C., Chu, C.Y., Rogers, M.S., Choy, K.W.,
and Pang, C.P., Journal of chromatography, 810, 187 (2004).
45. Manach, C., and Donovan, J.L., Free radical research, 38, 771
(2004).
46. Gennaro, L. et al., Journal of agricultural and food chemistry 50,
1904 (2002).
47. Nemeth, K., and Piskula, M.K., Critical reviews in food science
and nutrition, 47, 397 (2007).
48. Hollman, P.C., Van Het Hof, K.H., Tijburg, L.B., and Katan,
M.B., Free radical research, 34, 297 (2001).
49. Ness, A.R., and Powles, J.W., Int. J. Epidemiol, 26, 1 (1997).
50. Ferrali, M., Signorini, C., Caciotti, B., Sugherini, L., Ciccoli, L.,
Giachetti, D., and Comproti, M., FEBS Letters, 416, 123 (1997).
51. Elliott, A.J., Scheiber, S.A., Thomas, C., and Pardini, R.S.,
Biochem. Pharmacol, 44, 1603 (1992).
52. Hirano, R., Sasamoto, W., Matsumoto, A., Itakura, H., Igarashi,
O., and Kondo, K., J. Nutr. Sci. Vitaminol (Tokyo), 47, 357 (2001).
53. Cos, P., Ying, L., Calomme, M., Hu, J.P., Cimanga, K., Van Poel,
B., Pieters, L., Vlietinck, A.J., and Vanden Berghe, D., J. Nat.
Prod., 61, 71 (1998).
54. Nijveldt, R.J., van Nood, E., van Hoorn, D.E., Boelens, P.G., van
Norren, K., and van Leeuwen, P.A., Am. J. Clin. Nutr., 74, 418
(2001).
55. Halliwell, B., Lancet, 344, 721 (1994).
56. Muller, F.L., Lustgarten, M.S., Jang, Y., Richardson, A., and Van
Remmen, H., Free radical biology & medicine, 43, 477 (2007).
57. Valko, M., Izakovi, M., Mazu, M., Rhodes, C.J., and Telser, J.,
Molecular and cellular biochemistry, 266, 37 (2004).
Journal of Pharmacy Research Vol.5 Issue 8.August 2012
58. Nagy, I.Z., Annals of the NewYork Academy of Sciences, 928, 187
(2001).
59. Halliwell, B., The Biochemical journal, 401, 1 (2007).
60. Sohal, R.S., and Weindruch, R., Science, 273, 59 (1996).
61. Halliwell, B., Nutrition reviews, 55, 44 (1997).
62. Seifried, H.E., Anderson, D.E., Fisher, E.I., and Milner, J.A., J.
Nutr. Biochem., 18, 567 (2007).
63. Scalbert, A., Johnson, I.T., and Saltmarsh, M., The American
journal of clinical nutrition, 81, 215 (2005).
64. Bors, W., Heller, W., Michel, C., and Saran, M., Methods in
enzymology, 186, 343 (1990).
65. Jovanovic, S.V., and Simic, M.G., Annals of the New York Acad-
emy of Sciences, 899, 326 (2000).
66. Pietta, P.G., Journal of natural products, 63, 1035 (2000).
67. Mira, L., Fernandez, M.T., Santos, M., Rocha, R., Florencio,
M.H., and Jennings, K.R., Free radical research, 36, 1199 (2002).
68. Van Hoorn, D.E. et al., European journal of pharmacology, 45,
111 (2002).
69. Van Zanden, J.J., Geraets, L., Wortelboer, H.M., van Bladeren,
P.J., Rietjens, I.M., and Cnubben, N.H., Biochemical pharmacol-
ogy, 67, 1607 (2004).
70. Raso, G.M., Meli, R., Di Carlo, G., Pacilio, M., and Di Carlo, R.,
Life sciences, 68, 921 (2001).
71. Morre, D.J., Bridge, A., Wu, L.Y., and Morre, D.M., Biochemical
pharmacology, 60, 937 (2000).
72. Cao, G., Sofic, E., and Prior, R.L., Free radical biology & medi-
cine, 22, 749 (1997).
73. Michels, G., Haenen, G.R., Watjen, W., Rietjens, S., and Bast, A.,
Toxicology letters, 151, 105 (2004).
74. Hodnick, W.F., Kung, F.S., Roettger, W.J., Bohmont, C.W., and
Pardini, R.S., Biochemical pharmacology, 35, 2345 (1986).
75. Metodiewa, D., Jaiswal, A.K., Cenas, N., Dickancaite, E., and
Segura-Aguilar, J., Free radical biology & medicine, 26, 107 (1999).
76. Canada, A.T., Giannella, E., Nguyen, T.D., Free radical biology
& medicine, 9, 441 (1990).
77. Shao, Z.H., Vanden Hoek, T.L., Xie, J. et al., Cardiovascular
toxicology, 3, 331 (2003).
78. Azam, S., Hadi, N., Khan, N.U., and Hadi, S.M., Toxicol In Vitro
18, 555 (2004).
79. Raza, H., and John, A., Toxicol Appl Pharmacol, 207, 212 (2005).
80. Choi, E.J., European journal of pharmacology, 542, 162 (2006).
81. Woo, A.Y., Cheng, C.H., and Waye, M.M., Cardiovascular re-
search, 65, 244 (2005).
82. Bennets, H.W., Underwood F.L., and Shier, A., Aust. Vet. J., 22,
124 (1946).
83. Kuiper, G.G., Lemmen, J.G., and Carlsson, B., et al., Endocrinol-
ogy, 139, 4252 (1998).
84. Hackett, J.C., Kim, Y.W., Su, B., and Brueggemeier R.W., Bioor-
ganic & medicinal chemistry, 13, 4063 (2005).
85. Kao, Y.C., Zhou, C., Sherman, M., Laughton, C.A., and Chen, S.,
Environmental health perspectives, 106, 85 (1998).
86. Basly, J.P., and Lavier, M.C., Planta medica, 71, 287 (2005).
87. Pino, A.M., Valladares, L.E., Palma, M.A., Mancilla, A.M., Yanez,
M., and Albala, C., The Journal of clinical endocrinology and
metabolism, 85, 2797 (2000).
88. Cassidy, A., Bingham, S., and Setchell, K.D., The American jour-
nal of clinical nutrition, 60, 333 (2005).
89. Jakes, R.W., Alexander, L., Duffy, S.W., Leong, J., Chen, L.H.,
and Lee, W.H., Public health nutrition, 4, 191 (2001).
90. Kurzer, M.S., J. Nutr., 132, 570S (2002).
91. Beck, V., Unterrieder, E., Krenn, L., Kubelka, W., and Jungbauer,
4013-4020
 Kutaiba Ibrahim Alzand et al. / Journal of Pharmacy Research 2012,5(8),4013-4020
A., The Journal of steroid biochemistry and molecular biology, 96. Gilad, L.A., Tirosh, O., and Schwartz, B., The Journal of endo-
84, 259 (2003). crinology, 191, 387 (2006).
92. Fang, H., Tong, W., Branham, W.S. et al., Chemical research in 97. Farhan, H., Wahala, K., and Cross, H.S., The Journal of steroid
toxicology, 16, 1338 (2003). biochemistry and molecular biology, 84, 423 (2003).
93. Ricketts, M.L., Moore, D.D., Banz, W.J., Mezei, O., and Shay, 98. Szkudelska, K., and Nogowski, L., The Journal of steroid bio-
N.F., A review. J. Nutr. Biochem., 16, 321 (2005). chemistry and molecular biology, 105, 37 (2007).
94. Zand, R.S., Jenkins, D.J., and Diamandis, E.P., Breast cancer 99. Doerge, D.R., and Sheehan, D.M., Environmental health per-
research and treatment, 62, 35 (2000). spectives, 110, 349 (2002).
95. Rosenberg Zand, R.S., Jenkins, D.J., Brown, T.J., and Diamandis, 100. Hamilton-Reeves, J.M., Rebello, S.A., Thomas, W., Slaton, J.W.,
E.P., Clinica chimica acta; international journal of clinical chem- and Kurzer, M.S., J. Nutr., 137, 1769 (2005).
istry, 317, 17 (2002).
Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.5 Issue 8.August 2012 4013-4020